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Sample records for chromogenic differential medium

  1. Prospective evaluation of the chromogenic medium CandiSelect 4 for differentiation and presumptive identification of non-Candida albicans Candida species

    NARCIS (Netherlands)

    Zhao, Liang; de Hoog, G Sybren; Cornelissen, Akke; Lyu, Qian; Mou, Lili; Liu, Taohua; Cao, Yu; Vatanshenassan, Mansoureh; Kang, Yingqian

    Rapid identification of pathogenic yeasts is a crucial step in timely and appropriate antifungal therapy. For diagnostics in the clinical laboratory, simplified alternatives to barcoding are needed. CandiSelect 4 (CS4) medium, a chromogenic medium for isolation of clinical yeasts, allows routine

  2. Comparative evaluation of Strepto B ID chromogenic medium and Granada media for the detection of Group B streptococcus from vaginal samples of pregnant women.

    Science.gov (United States)

    Tazi, Asmaa; Réglier-Poupet, Hélène; Dautezac, François; Raymond, Josette; Poyart, Claire

    2008-06-01

    Two types of selective media, the chromogenic medium Strepto B ID and two non-chromogenic media Strepto B agar and the Granada medium, were tested and compared to blood agar plates (BAP) for screening of Group B streptococcus vaginal colonization in pregnant women. All tested media were comparable in terms of sensitivity however, their use in routine laboratories may markedly facilitate the rapid detection of GBS in vaginal samples.

  3. Evaluation of a Chromogenic Culture Medium for Isolation of Clostridium difficile within 24 Hours ▿

    Science.gov (United States)

    Perry, John D.; Asir, Kerry; Halimi, Diane; Orenga, Sylvain; Dale, Joanne; Payne, Michelle; Carlton, Ruth; Evans, Jim; Gould, F. Kate

    2010-01-01

    Rapid and effective methods for the isolation of Clostridium difficile from stool samples are desirable to obtain isolates for typing or to facilitate accurate diagnosis of C. difficile-associated diarrhea. We report on the evaluation of a prototype chromogenic medium (ID C. difficile prototype [IDCd]) for isolation of C. difficile. The chromogenic medium was compared using (i) 368 untreated stool samples that were also inoculated onto CLO medium, (ii) 339 stool samples that were subjected to alcohol shock and also inoculated onto five distinct selective agars, and (iii) standardized suspensions of 10 C. difficile ribotypes (untreated and alcohol treated) that were also inoculated onto five distinct selective agars. Two hundred thirty-six isolates of C. difficile were recovered from 368 untreated stool samples, and all but 1 of these strains (99.6%) were recovered on IDCd within 24 h, whereas 74.6% of isolates were recovered on CLO medium after 48 h. Of 339 alcohol-treated stool samples cultured onto IDCd and five other selective agars, C. difficile was recovered from 218 samples using a combination of all media. The use of IDCd allowed recovery of 96.3% of isolates within 24 h, whereas 51 to 83% of isolates were recovered within 24 h using the five other media. Finally, when they were challenged with pure cultures, all 10 ribotypes of C. difficile generated higher colony counts on IDCd irrespective of alcohol pretreatment or duration of incubation. We conclude that IDCd is an effective medium for isolation of C. difficile from stool samples within 24 h. PMID:20739493

  4. Chromogenic media for urine cultures can be cost-effective

    Directory of Open Access Journals (Sweden)

    Matjaž J. Retelj

    2007-03-01

    Full Text Available Background: Chromogenic media for diagnostic urinary bacteriology have several advantages over traditional media, such as cysteine-lactose-electrolyte deficient (CLED medium. Chromogenic media allow for easier recognition of mixed growth, save time, reduce workload and provide higher detection rates. However, the cost of chromogenic media is significantly higher compared to CLED and performance of chromogenic media varies depending on the manufacturer. In the present study, performance, turn-around time and cost of Uriselect4 chromogenic medium was compared to CLED.Methods: For performance analysis, 351 midstream urine (MSU samples from September 2005 to December 2005 were directly plated in parallel on Uriselect4 and CLED agar using the calibrated loop technique. Isolates on Uriselect4 were presumptively identified according to the product insert. For cost-effectiveness analysis, we included 1,972 consecutive MSU samples from May 2005 to July 2006. We compared the cost of required materials as well as technologists’ or specialists’ time for each medium examined.Results: No significant differences were found between the isolation rates of urinary pathogens on the studied media. The procedure using chromogenic media for uropathogens is slightly cheaper than the procedure using CLED, considering the proportion of bacteriuria positive samples (50.5 % and the distribution of taxa among isolates (namely Escherichia coli with 59.6 % observed in our laboratory. At the current isolation proportion in MSU samples processed in our laboratory, the average time to reporting results could be decreased by 0.3 days.Conclusions: Use of chromogenic media for urine investigations offers multiple advantages without increasing costs compared to procedures using CLED.

  5. Detection of oxacillin-susceptible mecA-positive Staphylococcus aureus isolates by use of chromogenic medium MRSA ID.

    Science.gov (United States)

    Kumar, V Anil; Steffy, Katherin; Chatterjee, Maitrayee; Sugumar, Madhan; Dinesh, Kavitha R; Manoharan, Anand; Karim, Shamsul; Biswas, Raja

    2013-01-01

    Reports of oxacillin-susceptible mecA-positive Staphylococcus aureus strains are on the rise. Because of their susceptibility to oxacillin and cefoxitin, it is very difficult to detect them by using routine phenotypic methods. We describe two such isolates that were detected by chromogenic medium and confirmed by characterization of the mecA gene element.

  6. CHROMagar COL-APSE: a selective bacterial culture medium for the isolation and differentiation of colistin-resistant Gram-negative pathogens

    DEFF Research Database (Denmark)

    Abdul Momin, Muhd Haziq F; Bean, David C; Hendriksen, Rene S.

    2017-01-01

    Purpose. A selective chromogenic culture medium for the laboratory isolation and differentiation of colistin resistant Acinetobacter, Pseudomonas, Stenotrophomonas and Enterobacteriaceae spp. (CHROMagar COL-APSE) was developed, evaluated and compared to an existing selective bacterial culture......-resistant non-fermentative bacteria (Acinetobacter, Pseudomonas and Stenotrophomonas). CHROMagar COL-APSE was also more sensitive in supporting the growth of Enterobacteriaceae with COL resistance associated with the carriage of mcr-1. Conclusion. CHROMagar COL-APSE is a sensitive and specific medium...

  7. Evaluation of chromogenic media and seminested PCR in the identification of Candida species

    Science.gov (United States)

    Daef, Enas; Moharram, Ahmed; Eldin, Salwa Seif; Elsherbiny, Nahla; Mohammed, Mona

    2014-01-01

    Identification of Candida cultured from various clinical specimens to the species level is increasingly necessary for clinical laboratories. Although sn PCR identifies the species within hours but its cost-effectiveness is to be considered. So there is always a need for media which help in the isolation and identification at the species level. The study aimed to evaluate the performance of different chromogenic media and to compare the effectiveness of the traditional phenotypic methods vs. seminested polymerase chain reaction (sn PCR) for identification of Candida species. One hundred and twenty seven Candida strains isolated from various clinical specimens were identified by conventional methods, four different chromogenic media and sn PCR. HiCrome Candida Differential and CHROMagar Candida media showed comparably high sensitivities and specificities in the identification of C. albicans, C. tropicalis, C. glabrata and C. krusei. CHROMagar Candida had an extra advantage of identifying all C. parapsilosis isolates. CHROMagar-Pal’s medium identified C. albicans, C. tropicalis and C. krusei with high sensitivities and specificities, but couldn’t identify C. glabrata or C. parapsilosis. It was the only medium that identified C. dubliniensis with a sensitivity and specificity of 100%. Biggy agar showed the least sensitivities and specificities. The overall concordance of the snPCR compared to the conventional tests including CHROMAgar Candida in the identification of Candida species was 97.5%. The use of CHROMAgar Candida medium is an easy and accurate method for presumptive identification of the most commonly encountered Candida spp. PMID:24948942

  8. Performance of chromogenic media for Candida in rapid presumptive identification of Candida species from clinical materials

    OpenAIRE

    Pravin Charles, M. V.; Kali, Arunava; Joseph, Noyal Mariya

    2015-01-01

    Background: In perspective of the worldwide increase in a number of immunocompromised patients, the need for identification of Candida species has become a major concern. The development of chromogenic differential media, introduced recently, facilitate rapid speciation. However, it can be employed for routine mycology workup only after an exhaustive evaluation of its benefit and cost effectiveness. This study was undertaken to evaluate the benefit and cost effectiveness of chromogenic media ...

  9. Exudative epidermitis in pigs caused by toxigenic Staphylococcus chromogenes.

    Science.gov (United States)

    Andresen, Lars Ole; Ahrens, Peter; Daugaard, Lise; Bille-Hansen, Vivi

    2005-02-25

    Staphylococcus chromogenes is closely related to Staphylococcus hyicus, which is recognised as the causative agent of exudative epidermitis (EE) in pigs. S. chromogenes is part of the normal skin flora of pigs, cattle and poultry and has so far been considered non-pathogenic to pigs. A strain of S. chromogenes producing exfoliative toxin type B, ExhB, was identified by the use of a multiplex PCR specific for the exfoliative toxins from S. hyicus. The exfoliative toxin from S. chromogenes reacted in immunoblot analysis with polyclonal and monoclonal antibodies specific to ExhB from S. hyicus and had an apparent molecular weight of 30 kDa. Sequencing the gene encoding the exfoliative toxin from S. chromogenes revealed that the molecular weight of the toxin with the signal peptide and the mature toxin was 30,553 and 26,694 Da, respectively. Comparison of the exhB genes from S. chromogenes strain VA654 and S. hyicus strain 1289D-88 showed differences in seven base pairs of the DNA sequences and in two amino acid residues in the deduced amino acid sequences. Pigs were experimentally inoculated with S. chromogenes strain VA654. By clinical observations and histopathological evaluation of the skin alterations, all pigs revealed development of generalized exudative epidermitis. No toxin producing S. hyicus was isolated from the pigs and all ExhB-positive bacterial isolates were identified as S. chromogenes. This confirmed that the disease-causing agent was the inoculated S. chromogenes strain VA654. The results of this study show that S. chromogenes may cause exudative epidermitis in pigs.

  10. Exudative epidermitis in pigs caused by toxigenic Staphylococcus chromogenes

    DEFF Research Database (Denmark)

    Andresen, Lars Ole; Ahrens, Peter; Daugaard, Lise

    2005-01-01

    Staphylococcus chromogenes is closely related to Staphylococcus hyicus, which is recognised as the causative agent of exudative epidermitis (EE) in pigs. S. chromogenes is part of the normal skin flora of pigs, cattle and poultry and has so far been considered non-pathogenic to pigs. A strain of S....... chromogenes producing exfoliative toxin type B, ExhB, was identified by the use of a multiplex PCR specific for the exfoliative toxins from S. hyicus. The exfoliative toxin from S. chromogenes reacted in immunoblot analysis with polyclonal and monoclonal antibodies specific to ExhB from S. hyicus and had...... an apparent molecular weight of 30 kDa. Sequencing the gene encoding the exfoliative toxin from S. chromogenes revealed that the molecular weight of the toxin with the signal peptide and the mature toxin was 30,553 and 26,694 Da, respectively. Comparison of the exhB genes from S. chromogenes strain VA654...

  11. Inhibitory effect of chromogenic culture media on the growth of Rhodotorula: relevance to the diagnosis of Rhodotorula spp. infections.

    Science.gov (United States)

    Bellanger, Anne-Pauline; Grenouillet, Frédéric; François, Nadine; Skana, Florence; Millon, Laurence

    2013-11-01

    With the increasing incidence and diverse etiologies of fungal infections, chromogenic yeast culture media are increasingly used for routine diagnosis. Rhodotorula species, which are characterized by the production of carotenoid pigments, are considered as emerging opportunistic pathogens. We recently diagnosed two fungemia due to Rhodotorula spp. and noticed that in both cases, the yeast failed to grow in subculture on the chromogenic yeast culture medium. This study was thus undertaken to investigate more thoroughly the ability (or inability) of Rhodotorula species to grow on different commercially available chromogenic media for yeast. Eighteen Rhodotorula spp. were checked for their ability to grow on four chromogenic yeast culture media: CHROMagar Candida (BD), Candi 4 Select (Biorad), Brilliance Candida (Oxoid), and Candida ID 2 (BioMerieux). All the Rhodotorula spp. strains grew on Brilliance and Candida ID 2, while only six isolates grew on Candi 4, and seven on CHROMagar. Two chromogenic yeast culture media showed a significant inhibitory effect on the growth of Rhodotorula species. As all Rhodotorula species are resistant to echinocandins and fluconazole, it is essential to isolate and identify these yeast quickly to initiate appropriate amphotericin B antifungal treatment as early as possible. The choice of media for routine use should take into account the ability of different media to allow all emerging fungal pathogens to grow. © 2013 APMIS. Published by John Wiley & Sons Ltd.

  12. Staphylococcus chromogenes, a Coagulase-Negative Staphylococcus Species That Can Clot Plasma

    OpenAIRE

    dos Santos, Danielle Cabral; Lange, Carla Christine; Avellar-Costa, Pedro; dos Santos, Katia Regina Netto; Brito, Maria Aparecida Vasconcelos Paiva; Giambiagi-deMarval, Marcia

    2016-01-01

    Staphylococcus chromogenes is one of the main coagulase-negative staphylococci isolated from mastitis of dairy cows. We describe S. chromogenes isolates that can clot plasma. Since the main pathogen causing mastitis is coagulase-positive Staphylococcus aureus, the coagulase-positive phenotype of S. chromogenes described here can easily lead to misidentification.

  13. Staphylococcus chromogenes, a Coagulase-Negative Staphylococcus Species That Can Clot Plasma

    Science.gov (United States)

    dos Santos, Danielle Cabral; Lange, Carla Christine; Avellar-Costa, Pedro; dos Santos, Katia Regina Netto; Brito, Maria Aparecida Vasconcelos Paiva

    2016-01-01

    Staphylococcus chromogenes is one of the main coagulase-negative staphylococci isolated from mastitis of dairy cows. We describe S. chromogenes isolates that can clot plasma. Since the main pathogen causing mastitis is coagulase-positive Staphylococcus aureus, the coagulase-positive phenotype of S. chromogenes described here can easily lead to misidentification. PMID:26912749

  14. Staphylococcus chromogenes, a Coagulase-Negative Staphylococcus Species That Can Clot Plasma.

    Science.gov (United States)

    Dos Santos, Danielle Cabral; Lange, Carla Christine; Avellar-Costa, Pedro; Dos Santos, Katia Regina Netto; Brito, Maria Aparecida Vasconcelos Paiva; Giambiagi-deMarval, Marcia

    2016-05-01

    Staphylococcus chromogenes is one of the main coagulase-negative staphylococci isolated from mastitis of dairy cows. We describe S. chromogenes isolates that can clot plasma. Since the main pathogen causing mastitis is coagulase-positive Staphylococcus aureus, the coagulase-positive phenotype of S. chromogenes described here can easily lead to misidentification. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  15. A selective and differential medium for Vibrio harveyi.

    Science.gov (United States)

    Harris, L; Owens, L; Smith, S

    1996-01-01

    A new medium, termed Vibrio harveyi agar, has been developed for the isolation and enumeration of V. harveyi. It is possible to differentiate V. harveyi colonies from the colonies of strains representing 15 other Vibrio species with this medium. This medium has been shown to inhibit the growth of two strains of marine Pseudomonas spp. and two strains of marine Flavobacterium spp. but to allow the growth of Photobacterium strains. Colonies displaying typical V. harveyi morphology were isolated from the larval rearing water of a commercial prawn hatchery with V. harveyi agar as a primary isolation medium and were positively identified, by conventional tests, as V. harveyi. This agar displays great potential as a primary isolation medium and offers significant advantages over thiosulfate-citrate-bile salts-sucrose agar as a medium for differentiating V. harveyi from other marine and estuarine Vibrio species. PMID:8795252

  16. A redox-mediated chromogenic reaction and application in immunoassay.

    Science.gov (United States)

    Yu, Ru-Jia; Ma, Wei; Peng, Mao-Pan; Bai, Zhi-Shan; Long, Yi-Tao

    2016-08-31

    A novel redox-mediated chromogenic reaction was demonstrated based on the reaction between HAuCl4 and 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), which generate various color responses from red to green in the resulting solutions. Various redox substance could be used to mediate the reaction and trigger a distinct color response. We established a sensitive hydrogen peroxide colorimetric sensor based on the redox-mediated chromogenic reaction and depicted the application both in detection of enzyme and in an immunoassay. Combining the traditional chromogenic reagent with gold nanoparticles, our assay has the advantage in short response time (within three minutes), high sensitivity (10(-12) g mL(-1) for HBsAg) and stability. Copyright © 2016. Published by Elsevier B.V.

  17. Development of selective and differential medium for Shigella sonnei using three carbohydrates (lactose, sorbitol, and xylose) and X-Gal.

    Science.gov (United States)

    Na, G N; Kim, S A; Kwon, O C; Rhee, M S

    2015-08-01

    The aim of this study was to develop a new selective and differential medium for isolating Shigella sonnei (designated 3SD medium). The new medium was based on three carbohydrates (lactose, sorbitol, and xylose) and a chromogenic substrate (5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside, X-Gal). S. sonnei cannot ferment lactose, sorbitol, or xylose, but can ferment X-Gal, which generates turquoise-blue colonies with rough edges. Other bacteria (54 strains of foodborne pathogens and spoilage bacteria) produced visually distinct colonies on 3SD medium (colorless or pink-violet colonies), or their growth was inhibited on 3SD medium. The optimum concentration of 50 mg/L X-Gal was selected because it yielded the highest level of morphological discrimination between S. sonnei and other bacteria, and this concentration was cost-effective. Bile salt concentration optimization was performed using healthy, heat-injured, and acid-injured S. sonnei. The recovery rate differed significantly depending on the bile salt concentration; media containing >1.0 g/L bile salt showed significantly lower recovery of stress-injured cells than medium containing 0.5 g/L bile salt (P<0.05). Growth of all Gram-positive bacteria was inhibited on medium containing 0.5 g/L bile salt; therefore, this concentration was used as the optimal concentration. Previous media used to isolate Shigella spp. (MacConkey, xylose lysine desoxycholate, and Salmonella-Shigella agar) showed poor performance when used to support the growth of injured S. sonnei cells, whereas 3SD medium supported a high growth rate of injured and healthy cells (equivalent to that obtained with nutrient-rich tryptic soy agar). To validate the performance of 3SD medium with real specimens, S. sonnei and other bacteria were spiked into samples such as untreated water, carrot, salad, and oyster. 3SD medium showed superior specificity (100%) and sensitivity (100%) for S. sonnei, and yielded no false-positive or false-negative results

  18. Comparison and Recovery of Escherichia coli and Thermotolerant Coliforms in Water with a Chromogenic Medium Incubated at 41 and 44.5°C

    Science.gov (United States)

    Alonso, Jose L.; Soriano, Adela; Carbajo, Oscar; Amoros, Inmaculada; Garelick, Hemda

    1999-01-01

    This study compared the performance of a commercial chromogenic medium, CHROMagarECC (CECC), and CECC supplemented with sodium pyruvate (CECCP) with the membrane filtration lauryl sulfate-based medium (mLSA) for enumeration of Escherichia coli and non-E. coli thermotolerant coliforms (KEC). To establish that we could recover the maximum KEC and E. coli population, we compared two incubation temperature regimens, 41 and 44.5°C. Statistical analysis by the Fisher test of data did not demonstrate any statistically significant differences (P = 0.05) in the enumeration of E. coli for the different media (CECC and CECCP) and incubation temperatures. Variance analysis of data performed on KEC counts showed significant differences (P = 0.01) between KEC counts at 41 and 44.5°C on both CECC and CECCP. Analysis of variance demonstrated statistically significant differences (P = 0.05) in the enumeration of total thermotolerant coliforms (TTCs) on CECC and CECCP compared with mLSA. Target colonies were confirmed to be E. coli at a rate of 91.5% and KEC of likely fecal origin at a rate of 77.4% when using CECCP incubated at 41°C. The results of this study showed that CECCP agar incubated at 41°C is efficient for the simultaneous enumeration of E. coli and KEC from river and marine waters. PMID:10427079

  19. Evaluation of chromogenic medium and direct latex agglutination test for detection of group B streptococcus in vaginal specimens from pregnant women in Lebanon and Kuwait.

    Science.gov (United States)

    Ghaddar, Nahed; Alfouzan, Wadha; Anastasiadis, Elie; Al Jiser, Tamima; Itani, Saad Eddine; Dernaika, Racha; Eid, Toufic; Ghaddar, Ali; Charafeddine, Adib; Dhar, Rita; El Hajj, Hiba

    2014-10-01

    This study was undertaken to evaluate chromogenic medium and a direct latex agglutination test (DLA) for detection of Group B Streptococcus (GBS) in the vaginal specimens of pregnant women, and to ascertain the prevalence of GBS in this population in Kuwait and Lebanon. Vaginal swabs, collected from women at 35-37 weeks of gestation, were cultured on 5 % sheep blood agar (SBA), colistin nalidixic acid agar (CNA), Strept B Select chromogenic agar (SBS) as well as Lim enrichment broth in 168 cases in Lebanon while only SBA was used for 1391 samples in Kuwait. In addition, vaginal samples from 102 GBS-positive and 20 GBS-negative women near the time of delivery were collected in Kuwait for evaluation of the DLA test. During the study period, the prevalence of GBS colonization was determined to be 20.7 % (288/1391) in Kuwait while 18.4 % (31) of 168 pregnant women in Lebanon had vaginal cultures positive for GBS. By direct plating of vaginal swabs on the three media used, the isolation rates of GBS were 51.6, 64.5 and 77.4 % on SBA, CNA and SBS, respectively, which increased to 90.35, 93.1 and 96.8 %, respectively, following subculture in Lim broth after 18 h of incubation. The sensitivity of the DLA test was found to be dependent on the density of GBS colonization, resulting in 100 % sensitivity and 100 % specificity for heavy (>10(2) c.f.u. per swab) and moderately heavy (50-100 c.f.u. per swab) growth of GBS. However, for vaginal specimens yielding women. The DLA test, in particular, could prove to be a useful tool for immediate detection of GBS in women near delivery so that intrapartum antibiotic prophylaxis can be initiated. © 2014 The Authors.

  20. A selective and differential medium for Vibrio harveyi.

    OpenAIRE

    Harris, L; Owens, L; Smith, S

    1996-01-01

    A new medium, termed Vibrio harveyi agar, has been developed for the isolation and enumeration of V. harveyi. It is possible to differentiate V. harveyi colonies from the colonies of strains representing 15 other Vibrio species with this medium. This medium has been shown to inhibit the growth of two strains of marine Pseudomonas spp. and two strains of marine Flavobacterium spp. but to allow the growth of Photobacterium strains. Colonies displaying typical V. harveyi morphology were isolated...

  1. Hyperspectral imaging for presumptive identification of bacterial colonies on solid chromogenic culture media

    Science.gov (United States)

    Guillemot, Mathilde; Midahuen, Rony; Archeny, Delpine; Fulchiron, Corine; Montvernay, Regis; Perrin, Guillaume; Leroux, Denis F.

    2016-04-01

    BioMérieux is automating the microbiology laboratory in order to reduce cost (less manpower and consumables), to improve performance (increased sensitivity, machine algorithms) and to gain traceability through optimization of the clinical laboratory workflow. In this study, we evaluate the potential of Hyperspectral imaging (HSI) as a substitute to human visual observation when performing the task of microbiological culture interpretation. Microbial colonies from 19 strains subcategorized in 6 chromogenic classes were analyzed after a 24h-growth on a chromogenic culture medium (chromID® CPS Elite, bioMérieux, France). The HSI analysis was performed in the VNIR region (400-900 nm) using a linescan configuration. Using algorithms relying on Linear Spectral Unmixing, and using exclusively Diffuse Reflectance Spectra (DRS) as input data, we report interclass classification accuracies of 100% using a fully automatable approach and no use of morphological information. In order to eventually simplify the instrument, the performance of degraded DRS was also evaluated using only the most discriminant 14 spectral channels (a model for a multispectral approach) or 3 channels (model of a RGB image). The overall classification performance remains unchanged for our multispectral model but is degraded for the predicted RGB model, hints that a multispectral solution might bring the answer for an improved colony recognition.

  2. Performance of chromogenic media for Candida in rapid presumptive identification of Candida species from clinical materials.

    Science.gov (United States)

    Pravin Charles, M V; Kali, Arunava; Joseph, Noyal Mariya

    2015-06-01

    In perspective of the worldwide increase in a number of immunocompromised patients, the need for identification of Candida species has become a major concern. The development of chromogenic differential media, introduced recently, facilitate rapid speciation. However, it can be employed for routine mycology workup only after an exhaustive evaluation of its benefit and cost effectiveness. This study was undertaken to evaluate the benefit and cost effectiveness of chromogenic media for speciation of Candida clinical isolates. Sputum samples of 382 patients were screened for the presence of Candida spp. by Gram stain and culture on sabouraud dextrose agar. Candida species were identified using Gram stain morphology, germ tube formation, cornmeal agar with Tween-80, sugar fermentation tests and morphology on HiCrome Candida differential agar. All the Candida isolates were inoculated on HiCrome Candida agar (HiMedia, Mumbai, India). The sensitivity and specificity of HiCrome agar for identification of Candida albicans were 90% and 96.42%, respectively whereas sensitivity and specificity of carbohydrate fermentation test were 86.67% and 74.07%, respectively. Sensitivity and specificity values of HiCrome agar for detection of C. albicans, Candida parapsilosis and Candida glabrata were above 90%. We found HiCrome agar has high sensitivity and specificity comparable to that of the conventional method. In addition, use of this differential media could significantly cut down the turnaround time as well as cost of sample processing.

  3. Structure elucidation of chromogen resulting from jaffe's reaction

    International Nuclear Information System (INIS)

    Hussain, I.; Tariq, M.I.; Siddiqui, H.L.

    2009-01-01

    Jaffe's reaction is a reaction of creatinine with alkaline picric acid resulting in red chromogen. This color is being used widely for the estimation of creatinine in biological fluids without knowing the exact structure of the chromogen. The red species is isolated and its UV/Visible, /sup 1/H- and /sup 13/C- NMR studies are carried out. It is found that the red species is not a pure compound but a mixture of stereoisomers. Moreover, the red species in water reflects the same UV /Visible spectra as given by the reaction of creatinine with alkaline picrate in aqueous media The red species is a 2:3 molar Meisenheimer a-complex of creatinine-picric acid. (author)

  4. Identification and antimicrobial susceptibility of Staphylococcus chromogenes isolates from intramammary infections of dairy cows.

    Science.gov (United States)

    Devriese, L A; Baele, M; Vaneechoutte, M; Martel, A; Haesebrouck, F

    2002-06-20

    Staphylococcus chromogenes is a highly prevalent species in subclinical mastitis with a well-established impact on somatic cell count. Few data are available on its antimicrobial susceptibility. The objective of this study was three-fold: (1) to evaluate simple identification tests by comparing them with a genomic method; (2) to determine minimal inhibitory concentrations (MICs) of different antibiotics; (3) to search for the presence of important resistance mechanisms and resistance-determining genes.Seventy-three staphylococcal strains, all collected on different dairy farms, were tentatively identified as S. chromogenes based on their lack of hemolysis and their characteristic intermediate DNase activity. The identification of 70 strains was confirmed as S. chromogenes by tRNA intergenic spacer PCR (tRNA PCR). Three strains were identified as S. sciuri, a species that is naturally cloxacillin- and lincomycin-resistant. All 70 S. chromogenes strains were found to be normally susceptible to neomycin, gentamicin, erythromycin, enrofloxacin, and to penicillinase-stable penicillins and cephalosporins, represented in this study by cloxacillin. The latter result was confirmed by the absence of the mecA gene in each of 13 strains in which this gene was searched for. Twenty-seven (38%) strains were penicillinase producers. Three lincomycin-resistant S. chromogenes strains were found to carry the linA gene. It was concluded that S. chromogenes can be identified reliably in routine mastitis bacteriology, and that the only resistance of importance is against penicillinase-susceptible penicillins.

  5. Local host response following an intramammary challenge with Staphylococcus fleurettii and different strains of Staphylococcus chromogenes in dairy heifers.

    Science.gov (United States)

    Piccart, Kristine; Verbeke, Joren; De Visscher, Anneleen; Piepers, Sofie; Haesebrouck, Freddy; De Vliegher, Sarne

    2016-05-12

    Coagulase-negative staphylococci (CNS) are a common cause of subclinical mastitis in dairy cattle. The CNS inhabit various ecological habitats, ranging between the environment and the host. In order to obtain a better insight into the host response, an experimental infection was carried out in eight healthy heifers in mid-lactation with three different CNS strains: a Staphylococcus fleurettii strain originating from sawdust bedding, an intramammary Staphylococcus chromogenes strain originating from a persistent intramammary infection (S. chromogenes IM) and a S. chromogenes strain isolated from a heifer's teat apex (S. chromogenes TA). Each heifer was inoculated in the mammary gland with 1.0 × 10(6) colony forming units of each bacterial strain (one strain per udder quarter), whereas the remaining quarter was infused with phosphate-buffered saline. Overall, the CNS evoked a mild local host response. The somatic cell count increased in all S. fleurettii-inoculated quarters, although the strain was eliminated within 12 h. The two S. chromogenes strains were shed in larger numbers for a longer period. Bacterial and somatic cell counts, as well as neutrophil responses, were higher after inoculation with S. chromogenes IM than with S. chromogenes TA. In conclusion, these results suggest that S. chromogenes might be better adapted to the mammary gland than S. fleurettii. Furthermore, not all S. chromogenes strains induce the same local host response.

  6. Comparative evaluation of six chromogenic media for presumptive yeast identification.

    Science.gov (United States)

    Vecchione, Alessandra; Florio, Walter; Celandroni, Francesco; Barnini, Simona; Lupetti, Antonella; Ghelardi, Emilia

    2017-12-01

    The present study was undertaken to evaluate the discrimination ability of six chromogenic media in presumptive yeast identification. We analysed 108 clinical isolates and reference strains belonging to eight different species: Candida albicans , Candida dubliniensis , Candida tropicalis , Candida krusei , Candida glabrata , Candida parapsilosis , Candida lusitaniae and Trichosporon mucoides . C. albicans , C. tropicalis and C. krusei could be distinguished from one another in all the tested chromogenic media, as predicted by the manufacturers. In addition, C. albicans could be distinguished from C. dubliniensis on BBL CHROMagar Candida, Kima CHROMagar Candida and Brilliance Candida, and C. parapsilosis could be identified on CHROMATIC Candida agar, CHROMOGENIC Candida agar, and Brilliance Candida agar. Brilliance Candida provided the widest discrimination ability, being able to discriminate five out of the seven Candida species tested. Interestingly, C. tropicalis and C. krusei could be already distinguished from each other after 24 hours of incubation. © Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2017. All rights reserved. No commercial use is permitted unless otherwise expressly granted.

  7. Pathogenesis and inflammatory response in experimental caprine mastitis due to Staphylococcus chromogenes.

    Science.gov (United States)

    Lasagno, M; Ortiz, M; Vissio, C; Yaciuk, R; Bonetto, C; Pellegrino, M; Bogni, C; Odierno, L; Raspanti, C

    2018-03-01

    Coagulase-negative staphylococci (CNS) are the most frequently isolated bacteria in cases of subclinical mastitis in dairy cows. CNS species may differ in their pathogenicity, but very little is known about their virulence factors or their immune response in intramammary infections. To our knowledge, no experimental studies into the mastitis pathogenesis caused by CNS have been described in lactating goats. The aim of this study was to induce an experimentally Staphylococcus chromogenes mastitis in lactating goats aimed at verifying if the model can be used to evaluate the inflammatory response, the dynamics of infection and the pathological findings within the first hours of intramammary inoculation. Six Saanen goats in mid-lactation were inoculated with 1 × 10 7 colony forming units of S. chromogenes. Bacterial growth peaked in milk from the challenged right halves of the mammary glands (RMG) at 4 h post inoculation (PI). Shedding of viable bacteria showed a marked decrease at 12 h PI. An increase in mean somatic cell counts was observed in the milk samples from 8 h PI onwards. Mild clinical signs were evoked by intramammary inoculation. Staphylococcus chromogenes could be isolated in tissue from all RMG. Histological examination of specimens of the RMG and lymph nodes of the goats showed an increased inflammatory response throughout the experiment with respect to control halves. In conclusion, the experimental inoculation of S. chromogenes in lactating goats is capable of eliciting an inflammatory response and capable of causing pathological changes. This research represents a preliminary study for a better knowledge of the mastitis pathogenesis caused by S. chromogenes. Copyright © 2018 Elsevier Ltd. All rights reserved.

  8. Differentiation of Dental Pulp Stem Cells into Neuron-Like Cells in Serum-Free Medium

    Directory of Open Access Journals (Sweden)

    Shahrul Hisham Zainal Ariffin

    2013-01-01

    Full Text Available Dental pulp tissue contains dental pulp stem cells (DPSCs. Dental pulp cells (also known as dental pulp-derived mesenchymal stem cells are capable of differentiating into multilineage cells including neuron-like cells. The aim of this study was to examine the capability of DPSCs to differentiate into neuron-like cells without using any reagents or growth factors. DPSCs were isolated from teeth extracted from 6- to 8-week-old mice and maintained in complete medium. The cells from the fourth passage were induced to differentiate by culturing in medium without serum or growth factors. RT-PCR molecular analysis showed characteristics of Cd146+, Cd166+, and Cd31− in DPSCs, indicating that these cells are mesenchymal stem cells rather than hematopoietic stem cells. After 5 days of neuronal differentiation, the cells showed neuron-like morphological changes and expressed MAP2 protein. The activation of Nestin was observed at low level prior to differentiation and increased after 5 days of culture in differentiation medium, whereas Tub3 was activated only after 5 days of neuronal differentiation. The proliferation of the differentiated cells decreased in comparison to that of the control cells. Dental pulp stem cells are induced to differentiate into neuron-like cells when cultured in serum- and growth factor-free medium.

  9. Yeast identification in routine clinical microbiology laboratory and its clinical relevance

    Directory of Open Access Journals (Sweden)

    S Agarwal

    2011-01-01

    Full Text Available Rapid identification of yeast infections is helpful in prompt appropriate antifungal therapy. In the present study, the usefulness of chromogenic medium, slide culture technique and Vitek2 Compact (V2C has been analysed. A total of 173 clinical isolates of yeast species were included in the study. An algorithm to identify such isolates in routine clinical microbiology laboratory was prepared and followed. Chromogenic medium was able to identify Candida albicans, C. tropicalis, C. krusei, C. parapsilosis and Trichosporon asahii. Chromogenic medium was also helpful in identifying "multi-species" yeast infections. The medium was unable to provide presumptive identification of C. pelliculosa, C. utilis, C. rugosa, C. glabrata and C. hemulonii. Vitek 2 compact (V2C differentiated all pseudohypae non-producing yeast species. The algorithm followed was helpful in timely presumptive identification and final diagnosis of yeast infections, including multi-species yeast infections.

  10. Complete chromogen separation and analysis in double immunohistochemical stains using Photoshop-based image analysis.

    Science.gov (United States)

    Lehr, H A; van der Loos, C M; Teeling, P; Gown, A M

    1999-01-01

    Simultaneous detection of two different antigens on paraffin-embedded and frozen tissues can be accomplished by double immunohistochemistry. However, many double chromogen systems suffer from signal overlap, precluding definite signal quantification. To separate and quantitatively analyze the different chromogens, we imported images into a Macintosh computer using a CCD camera attached to a diagnostic microscope and used Photoshop software for the recognition, selection, and separation of colors. We show here that Photoshop-based image analysis allows complete separation of chromogens not only on the basis of their RGB spectral characteristics, but also on the basis of information concerning saturation, hue, and luminosity intrinsic to the digitized images. We demonstrate that Photoshop-based image analysis provides superior results compared to color separation using bandpass filters. Quantification of the individual chromogens is then provided by Photoshop using the Histogram command, which supplies information on the luminosity (corresponding to gray levels of black-and-white images) and on the number of pixels as a measure of spatial distribution. (J Histochem Cytochem 47:119-125, 1999)

  11. In vitro growth inhibition of major mastitis pathogens by Staphylococcus chromogenes originating from teat apices of dairy heifers.

    Science.gov (United States)

    De Vliegher, S; Opsomer, G; Vanrolleghem, A; Devriese, L A; Sampimon, O C; Sol, J; Barkema, H W; Haesebrouck, F; de Kruif, A

    2004-07-14

    Earlier field observations suggest that teat apex colonization by Staphylococcus chromogenes pre-partum in dairy heifers protects udder quarters against elevated somatic cell counts early post-partum. To explain these findings, the in vitro inhibitory capability of S. chromogenes from teat apices of heifers towards some major mastitis pathogens was tested using a modified cross-streaking method. Two out of 10 S. chromogenes isolates, both originating from two different teats from the same heifer, consistently inhibited growth of all Staphylococcus aureus, Streptococcus dysgalactiae, and Streptococcus uberis strains, but none of the Escherichia coli strains. The present study, therefore, supports the protective effect of teat apex colonization by S. chromogenes by in vitro production of inhibitory substances.

  12. Prepartum teat apex colonization with Staphylococcus chromogenes in dairy heifers is associated with low somatic cell count in early lactation.

    Science.gov (United States)

    De Vliegher, S; Laevens, H; Devriese, L A; Opsomer, G; Leroy, J L M; Barkema, H W; de Kruif, A

    2003-04-02

    A high number of dairy heifers freshen with udder health problems. The prevalence of teat apex colonization (TAC) with Staphylococcus chromogenes, one of the most widespread coagulase-negative staphylococci (CNS) in milk samples from freshly calved dairy heifers, was measured cross-sectionally in non-lactating heifers on eight commercial dairy farms in Belgium. The influence of age on this prevalence, and the association between teat apex colonization with S. chromogenes prepartum and quarter milk somatic cell count (SCC) in early lactation were studied. In total, 492 teat apices were sampled from 123 heifers. The age of the heifers varied from 8 to 34 months. Overall, 20% of the heifers had at least one teat apex colonized with S. chromogenes. Of all teats sampled, 10% were colonized with S. chromogenes. The chance of having at least one teat apex colonized with S. chromogenes increased with age of the heifer. The presence of prepartum teat apex colonization with S. chromogenes was not associated with intramammary infection (IMI) early postpartum with the same bacterium. On the contrary, teat apex colonization with S. chromogenes prepartum appeared to protect quarters in the first few days of lactation from having somatic cell count >or=200000cells/ml milk, commonly accepted as the threshold for intramammary infection.

  13. Pulsed-field gel electrophoresis of Staphylococcus hyicus and Staphylococcus chromogenes genomic DNA and its taxonomic, epidemiologic and ecologic applications in veterinary medicine.

    Science.gov (United States)

    Shimizu, A; Kloos, W E; Berkhoff, H A; George, C G; Ballard, D N

    1997-06-01

    One hundred and thirty-eight strains of Staphylococcus hyicus and 21 strains of S. chromogenes isolated from animals were analyzed by pulsed-field gel electrophoresis (PFGE) after restriction endonuclease Smal digestion of chromosomal DNA. Eighty-eight strains of S. hyicus from pigs with or without exudative epidermitis (EE) generated 16 to 26 fragments in the size range of chromogenes from pigs and cows generated 17 to 24 fragments ranging from chromogenes strains were more highly conserved than those of S. hyicus. S. chromogenes strains could be distinguished from S. hyicus strains by fragments within the range of 305 to 545 kb. The results indicate that PFGE analysis could be used to distinguish between S. hyicus and S. chromogenes. We conclude that PFGE analysis is a useful tool not only for species or strain identification but also for epidemiologic or ecologic studies of S. hyicus and S. chromogenes.

  14. Chromogenic switchable glazing: Towards the development of the smart window

    Energy Technology Data Exchange (ETDEWEB)

    Lampert, C.M.

    1995-06-01

    The science and technology of chromogenic materials for switchable glazings in building applications is discussed. These glazings can be used for dynamic control of solar and visible energy. Currently many researchers and engineers are involved with the development of products in this field. A summary of activities in Japan, Europe, Australia, USA and Canada is made. The activities of the International Energy Agency are included. Both non-electrically activated and electrically activated glazings are discussed. Technologies covered in the first category are photochromics, and thermochromics and thermotropics. A discussion of electrically activated chromogenic glazings includes dispersed liquid crystals, dispersed particles and electrochromics. A selection of device structures and performance characteristics are compared. A discussion of transparent conductors is presented. Technical issues concerning large-area development of smart windows are discussed.

  15. A selective medium for the enumeration and differentiation of Lactobacillus delbrueckii ssp. bulgaricus.

    Science.gov (United States)

    Nwamaioha, Nwadiuto O; Ibrahim, Salam A

    2018-06-01

    Modified reinforced clostridial medium (mRCM) was developed and evaluated for the differential enumeration of Lactobacillus delbrueckii ssp. bulgaricus. Lactobacillus bulgaricus, an important species of lactic acid bacteria with health benefits, is used in the production of yogurt and other fermented foods. Our results showed that supplementing reinforced clostridial medium with 0.025% CaCl 2 , 0.01% uracil, and 0.2% Tween 80 (mRCM) significantly enhanced the growth rate of L. bulgaricus RR and ATCC 11842 strains as measured by the optical densities of these strains after 12 h of incubation at 42°C. The bacterial populations (plate count) of the RR and ATCC 11842 strains were 0.76 and 0.77 log cfu/g higher in mRCM than in de Man, Rogosa, and Sharpe and reinforced clostridial medium media, respectively. Conversely, the population counts for other bacterial species (Bifidobacterium, Lactobacillus rhamnosus, and Lactobacillus reuteri) were significantly inhibited in the mRCM medium. The addition of aniline blue dye to mRCM (mRCM-blue) improved the selectivity of L. bulgaricus in mixed lactic bacterial cultures compared with de Man, Rogosa, and Sharpe medium and lactic agar with regard to colony appearance and morphology. The mRCM-blue performed better than the conventional medium in culturing, enumerating, and differentiating L. bulgaricus. Therefore, mRCM-blue could be used as a selective medium to enhance the growth and differentiation of L. bulgaricus in order to meet the increasing demand for this beneficial species of bacteria. Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  16. Performance of Chromogenic Candida agar and CHROMagar Candida in recovery and presumptive identification of monofungal and polyfungal vaginal isolates.

    Science.gov (United States)

    Ozcan, Kadri; Ilkit, Macit; Ates, Aylin; Turac-Bicer, Aygul; Demirhindi, Hakan

    2010-02-01

    Chromogenic Candida agar (OCCA) is a novel medium facilitating isolation and identification of Candida albicans, C. tropicalis, and C. krusei, as well as indicating polyfungal population in clinical samples. We compare the performance of OCCA, to CHROMagar Candida (CAC) and Sabouraud chloramphenicol agar (SCA). Vaginal swab samples from 392 women were simultaneously inoculated onto three study media. A total of 161 (41.1%) were found to be positive for fungi of which 140 (87%) were monofungal, and 21 (13%) polyfungal. One-hundred and fifty-seven samples (97.5%) were positive on CAC, 156 (96.9%) on OCCA, 148 (91.9%) on SCA and 144 (89.4%) samples were positive on all three media. The yeasts were identified by conventional methods including germ tube test, microscopic morphology on cornmeal-Tween 80 agar, and the commercial API 20C AUX. The 182 isolates were C. albicans (n = 104), C. glabrata (n = 51), C. krusei (n = 7), C. tropicalis (n = 5), C. famata (n = 3), C. kefyr (n = 3), C. zeylanoides (n = 3), C. colliculosa (n = 2), and other species of Candida (n = 4). Among the 21 polyfungal populations, 20 (95.2%) were detected in OCCA, 14 (66.7%) in CAC, and 13 (61.9%) in CAC and OCCA (P or=92.9% at 72 h. OCCA is more efficient and reliable for rapidly identifying C. albicans and polyfungal populations than CAC. However, CAC is more efficient for identifying C. krusei and C. tropicalis. A chromogenic agar with a higher isolation rate of yeasts and better detection of polyfungal populations than SCA, is suggested as a medium of first choice when available.

  17. Draft Genome Sequence of Staphylococcus chromogenes Strain MU 970, Isolated from a Case of Chronic Bovine Mastitis

    OpenAIRE

    Fry, Pamela R.; Calcutt, Michael J.; Foecking, Mark F.; Hsieh, Hsin-Yeh; Suntrup, Douglas G.; Perry, Jeanette; Stewart, George C.; Middleton, John R.

    2014-01-01

    Coagulase-negative staphylococcal species are a common cause of subclinical bovine mastitis, with Staphylococcus chromogenes being one of the most frequently identified species in these cases. The draft genome sequence of an S. chromogenes isolate (MU 970) recovered from the milk of a cow with a chronic intramammary infection is reported here.

  18. Draft Genome Sequence of Staphylococcus chromogenes Strain MU 970, Isolated from a Case of Chronic Bovine Mastitis

    Science.gov (United States)

    Fry, Pamela R.; Calcutt, Michael J.; Foecking, Mark F.; Hsieh, Hsin-Yeh; Suntrup, Douglas G.; Perry, Jeanette; Stewart, George C.

    2014-01-01

    Coagulase-negative staphylococcal species are a common cause of subclinical bovine mastitis, with Staphylococcus chromogenes being one of the most frequently identified species in these cases. The draft genome sequence of an S. chromogenes isolate (MU 970) recovered from the milk of a cow with a chronic intramammary infection is reported here. PMID:25125652

  19. Draft Genome Sequence of Staphylococcus chromogenes Strain MU 970, Isolated from a Case of Chronic Bovine Mastitis.

    Science.gov (United States)

    Fry, Pamela R; Calcutt, Michael J; Foecking, Mark F; Hsieh, Hsin-Yeh; Suntrup, Douglas G; Perry, Jeanette; Stewart, George C; Middleton, John R

    2014-08-14

    Coagulase-negative staphylococcal species are a common cause of subclinical bovine mastitis, with Staphylococcus chromogenes being one of the most frequently identified species in these cases. The draft genome sequence of an S. chromogenes isolate (MU 970) recovered from the milk of a cow with a chronic intramammary infection is reported here. Copyright © 2014 Fry et al.

  20. Host response in bovine mastitis experimentally induced with Staphylococcus chromogenes.

    Science.gov (United States)

    Simojoki, H; Orro, T; Taponen, S; Pyörälä, S

    2009-02-16

    An experimental infection model was developed to study host response to intramammary infection in cows caused by Staphylococcus chromogenes. CNS intramammary infections have become very common in modern dairy herds, and they can remain persistent in the mammary gland. More information would be needed about the pathophysiology of CNS mastitis, and an experimental mastitis model is a means for this research. Six primiparous Holstein-Friesian cows were challenged with S. chromogenes 4 weeks after calving. One udder quarter of each cow was inoculated with 2.1 x 10(6)cfu of S. chromogenes. All cows became infected and clinical signs were mild. Milk production of the challenged quarter decreased on average by 16.3% during 7 days post-challenge. Cows eliminated bacteria in a few days, except for one cow which developed persistent mastitis. Milk indicators of inflammation, SCC and N-acetyl-beta-D-glucosaminidase (NAGase) returned to normal within a week. Milk NAGase activity increased moderately, which reflects minor tissue damage in the udder. Concentrations of serum amyloid A (SAA) and milk amyloid A (MAA) were both elevated at 12h PC. MAA was affected by the milking times, and was at its highest before the morning milking. In our experimental model, systemic acute phase protein response with SAA occurred as an on-off type reaction. In conclusion, this experimental model could be used to study host response in CNS mastitis caused by the main CNS species and also for comparison of the host response in a mild intramammary infection and in more severe mastitis models.

  1. Simple low cost differentiation of Candida auris from Candida haemulonii complex using CHROMagar Candida medium supplemented with Pal's medium.

    Science.gov (United States)

    Kumar, Anil; Sachu, Arun; Mohan, Karthika; Vinod, Vivek; Dinesh, Kavitha; Karim, Shamsul

    Candida auris is unique due to its multidrug resistance and misidentification as Candida haemulonii by commercial systems. Its correct identification is important to avoid inappropriate treatments. To develop a cheap method for differentiating C. auris from isolates identified as C. haemulonii by VITEK2. Fifteen C. auris isolates, six isolates each of C. haemulonii and Candida duobushaemulonii, and one isolate of Candida haemulonii var. vulnera were tested using CHROMagar Candida medium supplemented with Pal's agar for better differentiation. On CHROMagar Candida medium supplemented with Pal's agar all C. auris strains showed confluent growth of white to cream colored smooth colonies at 37°C and 42°C after 24 and 48h incubation and did not produce pseudohyphae. The isolates of the C. haemulonii complex, on the contrary, showed poor growth of smooth, light-pink colonies at 24h while at 48h the growth was semiconfluent with the production of pseudohyphae. C. haemulonii complex failed to grow at 42°C. We report a rapid and cheap method using CHROMagar Candida medium supplemented with Pal's agar for differentiating C. auris from isolates identified as C. haemulonii by VITEK2. Copyright © 2017 Asociación Española de Micología. Publicado por Elsevier España, S.L.U. All rights reserved.

  2. Chromogenic Factor VIII Assays for Improved Diagnosis of Hemophilia A.

    Science.gov (United States)

    Rodgers, Susan; Duncan, Elizabeth

    2017-01-01

    Hemophilia A is an inherited bleeding disorder caused by a reduced level of factor VIII coagulant activity (FVIII:C) in blood. Bleeding episodes may occur spontaneously in the severe form of hemophilia or after trauma in the milder forms. It is important that patients are diagnosed correctly, which includes placing them into the correct severity category of the disorder so that appropriate treatment can be given. Diagnosis is made by determination of the amount of FVIII:C in the blood, usually using a one-stage factor VIII:C assay. However, approximately one third of patients with mild or moderate hemophilia will have much lower results by the chromogenic assay, with some of them having normal results by the one-stage assay. The chromogenic factor VIII assay is used in some specialized hemophilia reference centers and is recommended for the diagnosis of mild hemophilia A, as this assay is considered to better reflect the severity status of hemophilia patients than the one-stage assay.

  3. Chromogenic assay for BAY 81-8973 potency assignment has no impact on clinical outcome or monitoring in patient samples.

    Science.gov (United States)

    Kitchen, S; Katterle, Y; Beckmann, H; Maas Enriquez, M

    2016-06-01

    Essentials Discrepancies can exist in factor VIII activity measured by the one-stage or chromogenic assays. LEOPOLD trial data were used to assess clinical impact of BAY 81-8973 potency assignment assay. Efficacy was not affected by the assay used for potency assignment and dosing of BAY 81-8973. Either assay may be used to measure factor VIII activity after BAY 81-8973 infusion. Background Product-specific discrepancies have been reported for factor VIII (FVIII) activity determined with one-stage or chromogenic assays. Objective To assess the clinical impact of potency assignment of BAY 81-8973, a full-length, unmodified, recombinant human FVIII, by use of the chromogenic assay or chromogenic assay adjusted to mimic results obtained with the one-stage assay Patients/methods Patients aged 12-65 years with severe hemophilia A received BAY 81-8973 in LEOPOLD I (20-50 IU kg(-1) two or three times weekly [investigator decision]) and LEOPOLD II (randomized to 20-30 IU kg(-1) twice weekly, 30-40 IU kg(-1) three times weekly, or on-demand treatment). Both trials included two 6-month crossover periods in which potency labeling was determined with the chromogenic substrate assay as per the European Pharmacopoeia (CS/EP) or the chromogenic substrate assay adjusted to mimic results obtained with the one-stage assay (CS/ADJ). The annualized bleeding rate (ABR) and FVIII incremental recovery were assessed by the use of pooled data. Results The analysis was perfomed on 121 patients. Median (quartile [Q] 1; Q3) ABRs during the CS/EP and CS/ADJ periods were 1.98 (0; 5.92) and 1.98 (0; 7.34), respectively. The mean incremental recovery was > 2 IU dL(-1) per IU kg(-1) in both periods with the use of either assay for postinfusion FVIII measurements. The median (Q1; Q3) chromogenic/one-stage assay recovery ratio was 1.054 (0.892; 1.150) for the CS/EP period when a plasma standard was used for calibration. Conclusions No impact on the ABR was observed with chromogenic-based as compared

  4. A differential medium for the enumeration of the spoilage yeast Zygosaccharomyces bailii in wine.

    Science.gov (United States)

    Schuller, D; Côrte-Real, M; Leão, C

    2000-11-01

    A collection of yeasts, isolated mostly from spoiled wines, was used in order to develop a differential medium for Zygosaccharomyces bailii. The 118 selected strains of 21 species differed in their origin and resistance to preservatives and belonged to the genera Pichia, Torulaspora, Dekkera, Debaryomyces, Saccharomycodes, Issatchenkia, Kluyveromyces, Kloeckera, Lodderomyces, Schizosaccharomyces, Rhodotorula, Saccharomyces, and Zygosaccharomyces. The design of the culture medium was based on the different ability of the various yeast species to grow in a mineral medium with glucose and formic acid (mixed-substrate medium) as the only carbon and energy sources and supplemented with an acid-base indicator. By manipulating the concentration of the acid and the sugar it was possible to select conditions where only Z. bailii strains gave rise to alkalinization, associated with a color change of the medium (positive response). The final composition of the mixed medium was adjusted as a compromise between the percentage of recovery and selectivity for Z. bailii. This was accomplished by the use of pure or mixed cultures of the yeast strains and applying the membrane filtration methodology. The microbiological analysis of two samples of contaminated Vinho Verde showed that the new medium can be considered as a differential medium to distinguish Z. bailii from other contaminating yeasts, having potential application in the microbiological control of wines and probably other beverages and foods.

  5. Quantitative digital image analysis of chromogenic assays for high throughput screening of alpha-amylase mutant libraries.

    Science.gov (United States)

    Shankar, Manoharan; Priyadharshini, Ramachandran; Gunasekaran, Paramasamy

    2009-08-01

    An image analysis-based method for high throughput screening of an alpha-amylase mutant library using chromogenic assays was developed. Assays were performed in microplates and high resolution images of the assay plates were read using the Virtual Microplate Reader (VMR) script to quantify the concentration of the chromogen. This method is fast and sensitive in quantifying 0.025-0.3 mg starch/ml as well as 0.05-0.75 mg glucose/ml. It was also an effective screening method for improved alpha-amylase activity with a coefficient of variance of 18%.

  6. Anionic chromogenic chemosensors highly selective for fluoride or cyanide based on 4-(4-Nitrobenzylideneamine)phenol

    Energy Technology Data Exchange (ETDEWEB)

    Nicoleti, Celso R; Marini, Vanderleia G; Zimmermann, Lizandra M; Machado, Vanderlei G., E-mail: vanderlei.machado@ufsc.br [Departamento de Quimica, Universidade Federal de Santa Catarina (UFSC), Florianopolis, SC (Brazil)

    2012-08-15

    4-(4-Nitrobenzylideneamine)phenol was used in two strategies allowing the highly selective detection of F{sup -} and CN{sup -}. Firstly, the compound in acetonitrile acts as a chromogenic chemosensor based on the idea that more basic anions cause its deprotonation (colorless solution), generating a colored solution containing phenolate. The discrimination of CN{sup -} over F{sup -} was obtained by adding 1.4% water to acetonitrile: water preferentially solvates F{sup -}, leaving the CN{sup -} free to deprotonate the compound. Another strategy involved an assay comprised of the competition between phenolate dye and the analyte for calyx[4]pyrrole in acetonitrile, a receptor highly selective for F{sup -}. Phenolate and calyx[4]pyrrole form a hydrogen-bonded complex, which changes the color of the medium. On the addition of various anions, only F{sup -} was able to restore the original color corresponding to phenolate in solution due to the fact that the anion dislodges phenolate from the complexation site. (author)

  7. Anionic chromogenic chemosensors highly selective for fluoride or cyanide based on 4-(4-Nitrobenzylideneamine)phenol

    International Nuclear Information System (INIS)

    Nicoleti, Celso R.; Marini, Vanderleia G.; Zimmermann, Lizandra M.; Machado, Vanderlei G.

    2012-01-01

    4-(4-Nitrobenzylideneamine)phenol was used in two strategies allowing the highly selective detection of F - and CN - . Firstly, the compound in acetonitrile acts as a chromogenic chemosensor based on the idea that more basic anions cause its deprotonation (colorless solution), generating a colored solution containing phenolate. The discrimination of CN - over F - was obtained by adding 1.4% water to acetonitrile: water preferentially solvates F - , leaving the CN - free to deprotonate the compound. Another strategy involved an assay comprised of the competition between phenolate dye and the analyte for calyx[4]pyrrole in acetonitrile, a receptor highly selective for F - . Phenolate and calyx[4]pyrrole form a hydrogen-bonded complex, which changes the color of the medium. On the addition of various anions, only F - was able to restore the original color corresponding to phenolate in solution due to the fact that the anion dislodges phenolate from the complexation site. (author)

  8. A Decade of Development of Chromogenic Culture Media for Clinical Microbiology in an Era of Molecular Diagnostics.

    Science.gov (United States)

    Perry, John D

    2017-04-01

    In the last 25 years, chromogenic culture media have found widespread application in diagnostic clinical microbiology. In the last decade, the range of media available to clinical laboratories has expanded greatly, allowing specific detection of additional pathogens, including Pseudomonas aeruginosa, group B streptococci, Clostridium difficile, Campylobacter spp., and Yersinia enterocolitica. New media have also been developed to screen for pathogens with acquired antimicrobial resistance, including vancomycin-resistant enterococci, carbapenem-resistant Acinetobacter spp., and Enterobacteriaceae with extended-spectrum β-lactamases and carbapenemases. This review seeks to explore the utility of chromogenic media in clinical microbiology, with particular attention given to media that have been commercialized in the last decade. The impact of laboratory automation and complementary technologies such as matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) is also assessed. Finally, the review also seeks to demarcate the role of chromogenic media in an era of molecular diagnostics. © Crown copyright 2017.

  9. Maintenance and neuronal cell differentiation of neural stem cells C17.2 correlated to medium availability sets design criteria in microfluidic systems.

    Directory of Open Access Journals (Sweden)

    Bu Wang

    Full Text Available BACKGROUND: Neural stem cells (NSCs play an important role in developing potential cell-based therapeutics for neurodegenerative disease. Microfluidics has proven a powerful tool in mechanistic studies of NSC differentiation. However, NSCs are prone to differentiate when the nutrients are limited, which occurs unfavorable by fast medium consumption in miniaturized culture environment. For mechanistic studies of NSCs in microfluidics, it is vital that neuronal cell differentiation is triggered by controlled factors only. Thus, we studied the correlation between available cell medium and spontaneous neuronal cell differentiation of C17.2 NSCs in standard culture medium, and proposed the necessary microfluidic design criteria to prevent undesirable cell phenotype changes. METHODOLOGY/PRINCIPAL FINDINGS: A series of microchannels with specific geometric parameters were designed to provide different amount of medium to the cells over time. A medium factor (MF, defined as the volume of stem cell culture medium divided by total number of cells at seeding and number of hours between medium replacement successfully correlated the amount of medium available to each cell averaged over time to neuronal cell differentiation. MF smaller than 8.3×10(4 µm3/cell⋅hour produced significant neuronal cell differentiation marked by cell morphological change and significantly more cells with positive β-tubulin-III and MAP2 staining than the control. When MF was equal or greater than 8.3×10(4 µm3/cell⋅hour, minimal spontaneous neuronal cell differentiation happened relative to the control. MF had minimal relation with the average neurite length. SIGNIFICANCE: MFs can be controlled easily to maintain the stem cell status of C17.2 NSCs or to induce spontaneous neuronal cell differentiation in standard stem cell culture medium. This finding is useful in designing microfluidic culture platforms for controllable NSC maintenance and differentiation. This study also

  10. The correlation between dual-color chromogenic in situ hybridization and fluorescence in situ hybridization in assessing HER2 gene amplification in breast cancer

    DEFF Research Database (Denmark)

    Pedersen, Marianne; Rasmussen, Birgitte Bruun

    2009-01-01

    and the generated chromogenic signals are also stable. This study presents a dual color CISH for simultaneous detection of the HER2 gene and chromosome 17. The CISH method performs a chromogenic detection "on top" of the Food and Drug Administration (FDA)-approved HER2 FISH pharmDx method, where the fluorochrome......-labeled probes are detected using enzyme-labeled antibodies and visualized by chromogenic enzymatic reactions. The HER2 status (amplified/not amplified and HER2 ratios) was evaluated by the CISH method and compared with results obtained by the FDA-approved FISH method. Of the 72 successfully investigated...

  11. Evaluation of Myc Gene Amplification in Prostate Cancer Using a Dual Color Chromogenic in-Situ Hybridization (Dual CISH) Assay

    OpenAIRE

    Daniel Lerda; Marta Cabrera; Jorge Flores; Luis Gutierrez; Armando Chierichetti; Martin Revol; Hernan Garcia Onto

    2013-01-01

    Objetive: The overall purpose of the study was to demonstrate applicability of the Dako dual-color chromogenic in situ hybridization (CISH) assay (DAKO Denmark, Glostrup) with respect to fluorescence in situ hybridization (FISH) probes MYC-C. Methods: MYC gene amplification by FISH and Dako dual-color CISH Results: The study showed that the dual-color CISH assay can convert Texas red and fluorescein isothiocyanate (FITC) signals into chromogenic signals. The dual –color CISH assay was p...

  12. A chromogenic and fluorogenic rhodol-based chemosensor for hydrazine detection and its application in live cell bioimaging

    Science.gov (United States)

    Tiensomjitr, Khomsan; Noorat, Rattha; Chomngam, Sinchai; Wechakorn, Kanokorn; Prabpai, Samran; Kanjanasirirat, Phongthon; Pewkliang, Yongyut; Borwornpinyo, Suparerk; Kongsaeree, Palangpon

    2018-04-01

    A rhodol-based fluorescent probe has been developed as a selective hydrazine chemosensor using levulinate as a recognition site. The rhodol levulinate probe (RL) demonstrated high selectivity and sensitivity toward hydrazine among other molecules. The chromogenic response of RL solution to hydrazine from colorless to pink could be readily observed by the naked eye, while strong fluorescence emission could be monitored upon excitation at 525 nm. The detection process occurred via a ring-opening process of the spirolactone initiated by hydrazinolysis, triggering the fluorescence emission with a 53-fold enhancement. The probe rapidly reacted with hydrazine in aqueous medium with the detection limit of 26 nM (0.83 ppb), lower than the threshold limit value (TLV) of 10 ppb suggested by the U.S. Environmental Protection Agency. Furthermore, RL-impregnated paper strips could detect hydrazine vapor. For biological applicability of RL, its membrane-permeable property led to bioimaging of hydrazine in live HepG2 cells by confocal fluorescence microscopy.

  13. Detection of c-myc amplification in formalin-fixed paraffin-embedded tumor tissue by chromogenic in situ hybridization (CISH).

    Science.gov (United States)

    Todorović-Raković, Nataša

    2013-01-01

    In situ hybridization (ISH) allows evaluation of genetic abnormalities, such as changes in chromosome number, chromosome translocations or gene amplifications, by hybridization of tagged DNA (or RNA) probes with complementary DNA (or RNA) sequences in interphase nuclei of target tissue. However, chromogenic in situ hybridization (CISH) is also applicable to formalin-fixed, paraffin-embedded (FFPE) tissues, besides metaphase chromosome spreads. CISH is similar to fluorescent in situ hybridization (FISH) regarding pretreatments and hybridization protocols but differs in the way of visualization. Indeed, CISH signal detection is similar to that used in immunohistochemistry, making use of a peroxidase-based chromogenic reaction instead of fluorescent dyes. In particular, tagged DNA probes are indirectly detected using an enzyme-conjugated antibody targeting the tags. The enzymatic reaction of the chromogenic substrate leads to the formation of strong permanent brown signals that can be visualized by bright-field microscopy at 40 × magnification. The advantage of CISH is that it allows the simultaneous observation of gene amplification and tissue morphology and the slides can be stored for a long time.

  14. Soft-tissue tumor differentiation using 3D power Doppler ultrasonography with echo-contrast medium injection.

    Science.gov (United States)

    Chiou, Hong-Jen; Chou, Yi-Hong; Chen, Wei-Ming; Chen, Winby; Wang, Hsin-Kai; Chang, Cheng-Yen

    2010-12-01

    We aimed to evaluate the ability of 3-dimensional power Doppler ultrasonography to differentiate soft-tissue masses from blood flow and vascularization with contrast medium. Twenty-five patients (mean age, 44.1 years; range, 12-77 years) with a palpable mass were enrolled in this study. Volume data were acquired using linear and convex 3-dimensional probes and contrast medium injected manually by bolus. Data were stored and traced slice by slice for 12 slices. All patients were scanned by the same senior sonologist. The vascular index (VI), flow index (FI), and vascular-flow index (VFI) were automatically calculated after the tumor was completely traced. All tumors were later confirmed by pathology. The study included 8 benign (mean, 36.5 mL; range, 2.4-124 mL) and 17 malignant (mean, 319.4 mL; range, 9.9-1,179.6 mL) tumors. Before contrast medium injection, mean VI, FI and VFI were, respectively, 3.22, 32.26 and 1.07 in benign tumors, and 1.97, 29.33 and 0.67 in malignant tumors. After contrast medium injection, they were, respectively, 20.85, 37.33 and 8.52 in benign tumors, and 40.12, 41.21 and 17.77 in malignant tumors. The mean differences between with and without contrast injection for VI, FI and VFI were, respectively, 17.63, 5.07 and 7.45 in benign tumors, and 38.15, 11.88 and 16.55 in malignant tumors. Tumor volume, VI, FI and VFI were not significantly different between benign and malignant tumors before and after echo-contrast medium injection. However, VI, FI and VFI under self-differentiation (differences between with and without contrast injection) were significantly different between malignant and benign tumors. Three-dimensional power Doppler ultrasound is a valuable tool for differential diagnosis of soft-tissue tumors, especially with the injection of an echo-contrast medium. Copyright © 2010 Elsevier. Published by Elsevier B.V. All rights reserved.

  15. Double-staining chromogenic in situ hybridization as a useful alternative to split-signal fluorescence in situ hybridization in lymphoma diagnostics

    DEFF Research Database (Denmark)

    van Rijk, A.; Svenstroup-Poulsen, T.; Jones, M.

    2010-01-01

    within the reach of every pathology laboratory. Design and Methods Our study was initiated to determine the consistency between chromogenic in situ hybridization and fluorescence in situ hybridization, both using split-signal probes developed for the detection of chromosomal breaks. Five hundred...... and actual signal were compared to the original fluorescence hybridization results. In addition, hematoxylin background staining intensity and signal intensity of the double-staining chromogenic in situ hybridization procedure were analyzed. Results With respect to the presence or absence of chromosomal...

  16. Differentiated strategy, business performance, and intellectual capital: Evidence small medium enterprise from Indonesia

    Science.gov (United States)

    Hariyati; Venusita, L.; Dyani, Z. F.

    2018-01-01

    Small and Medium Enterprises (SMEs) have a very important position in Indonesian economics. Implementation of the differentiated strategy has been impacted on improving the business performance of SMEs where the role of intellectual capital strongly supports the success of the implementation of the differentiated strategy. This study applied quantitative research which used survey method. This research examines the relationship between differentiated strategy to the performance of SMEs with mediated by intellectual capital. The results of this study show that intellectual capital mediates the relationship between differentiation strategies and business performance of SMEs. This study theoretically proves the importance of contextual variables in contingency theory. The practical results of this study contribute to raising awareness of business unit managers or other equivalent positions, especially managers in East Java to understand the importance of the role of intellectual capital, this is because intellectual capital meets the criteria as a unique source of the company that is able to create competitive advantage and increase the firm’s value.

  17. Evaluation of a Chromogenic Agar under Development To Screen for VRE Colonization ▿

    OpenAIRE

    Kallstrom, George; Doern, Christopher D.; Dunne, W. Michael

    2010-01-01

    BBL CHROMagar VanRE (CVRE) was compared with bile esculin azide agar plus vancomycin to screen for vancomycin-resistant enterococcus (VRE) colonization. CVRE distinguishes Enterococcus faecalis (green colonies) from Enterococcus faecium (mauve colonies) on the basis of chromogenic substrate use. CVRE sensitivity and specificity were 98.6% and 99.1%. Positive and negative predictive values were 95.9% and 99.7%.

  18. The importance of matrix-assisted laser desorption ionization–time of flight mass spectrometry for correct identification of Clostridium difficile isolated from chromID C. difficile chromogenic agar

    Directory of Open Access Journals (Sweden)

    Jonathan H.K. Chen

    2017-10-01

    Full Text Available The clinical workflow of using chromogenic agar and matrix-assisted laser desorption ionization time-of-fight mass spectrometry (MALDI-TOF MS for Clostridium difficile identification was evaluated. The addition of MALDI-TOF MS identification after the chromID C. difficile chromogenic agar culture could significantly improve the diagnostic accuracy of C. difficile.

  19. Dendrimeric tweezers for recognition of fluorogenic Co{sup 2+}, Mg{sup 2+} and chromogenic Fe{sup 2+}

    Energy Technology Data Exchange (ETDEWEB)

    Roy, Chandana B.; Meshram, Jyotsna S., E-mail: drjsmeshram@gmail.com

    2016-03-15

    Dendrimers are the attractive candidature for the formation of metal complexes capable of performing varied application, owing to the presence of multiple terminal groups on the exterior of the molecule has received tremendous attention. Herein, we have synthesized novel dendritic macromolecule (N′E,N‴E,N″‴E,N‴‴′E)-3,3′,3″,3‴-(ethane-1,2-diylbis(azanetriyle)) tetrakis(N'-(2-hydroxybenzyllidene)propanehydrazide) chemosensor L and its metal complexes. In the present study the application in the optical sensing for chromogenic Fe{sup 2+} and fluorogenic Co{sup 2+} and Mg{sup 2+}cation is reported. The dendrimeric chemosensor L and its metal complexes are investigated with the help of FTIR spectroscopy, Nuclear magnetic resonance ({sup 1}H NMR and {sup 13}C NMR), FT Raman Microspectroscopy, fluorescence and UV–visible spectroscopy. Thermal properties are studied using thermal gravimetric analysis. - Highlights: • Dual effect – Chromogenic and fluorogenic. Chemosensor shows chromogenic effect towards Fe{sup 2+} as well as fluorogenic effect towards Co{sup 2+}and Mg{sup 2+} cation. • From Linear fitting calibration plot for computing LOD and LOQ, it was detected that – LOD=32.3 nM, LOQ=97.8 nM. • Jobs Plot – A graph plotted [HG]={(ΔF/Fo)[H]} Vs {[H]v/([H]v+[G]v)} has maxima at 0.33 which corresponds to 1:2 stoichiometry of chemosensor L:Co{sup 2+}.

  20. Selective chromogenic detection of thiol-containing biomolecules using carbonaceous nanospheres loaded with silver nanoparticles as carrier.

    Science.gov (United States)

    Hu, Bo; Zhao, Yang; Zhu, Hai-Zhou; Yu, Shu-Hong

    2011-04-26

    Thiol-containing biomolecules show strong affinity with noble metal nanostructures and could not only stably protect them but also control the self-assembly process of these special nanostructures. A highly selective and sensitive chromogenic detection method has been designed for the low and high molecular weight thiol-containing biomolecules, including cysteine, glutathione, dithiothreitol, and bovine serum albumin, using a new type of carbonaceous nanospheres loaded with silver nanoparticles (Ag NPs) as carrier. This strategy relies upon the place-exchange process between the reporter dyes on the surface of Ag NPs and the thiol groups of thiol-containing biomolecules. The concentration of biomolecules can be determined by monitoring with the fluorescence intensity of reporter dyes dispersed in solution. This new chromogenic assay method could selectively detect these biomolecules in the presence of various other amino acids and monosaccharides and even sensitively detect the thiol-containing biomolecules with different molecular weight, even including proteins.

  1. Determination of HER-2 status on FNAC material from breast carcinomas using in situ hybridization with dual chromogen visualization with silver enhancement (dual SISH

    Directory of Open Access Journals (Sweden)

    Beraki Elsa

    2010-01-01

    Full Text Available During the last years, HER-2 status kits and protocols for chromogen visualization of hybridization signals have come on the market. The first generation using chromogen visualization used single color probes. The second generation, now emerging on the market, uses dual chromogen visualization. The aim of this study has been to test a new dual color chromogen kit (Ventana INFORM HER2 Dual Colour ISH Roche ® and compare the results with our in-house method(s. The material consisted primarily of cytological material from invasive breast carcinomas in 49 women. Dual SISH was done on all 49 cytological and histological specimens. The histological specimens were treated according to the manufacturer′s recommendations. The procedure was modified in several steps in order to adapt it to the cytological material. Hybridization failed in two cytological specimens. Dual SISH showed concordant results on cytological and histological material as to amplified/not amplified. The included cases had the same HER-2 expression in the invasive and the in situ components on histology. Four IDC showed HER-2 amplification (8.5%. Polysomy was found in two cases. All dual SISH results except for one concurred with the results of the in-house method(s (1/47=2.1%. The dual SISH is suitable for cytological examination of HER-2 status. The protocol must be optimized for cytological material.

  2. The correlation between dual-color chromogenic in situ hybridization and fluorescence in situ hybridization in assessing HER2 gene amplification in breast cancer.

    Science.gov (United States)

    Pedersen, Marianne; Rasmussen, Birgitte Bruun

    2009-06-01

    Fluorescence in situ hybridization (FISH) is regarded as the gold standard method for detecting HER2 gene amplification. Chromogenic in situ hybridization (CISH) is a promising alternative to FISH because CISH has the advantages of being a method evaluated by bright-field microscopy and the generated chromogenic signals are also stable. This study presents a dual color CISH for simultaneous detection of the HER2 gene and chromosome 17. The CISH method performs a chromogenic detection "on top" of the Food and Drug Administration (FDA)-approved HER2 FISH pharmDx method, where the fluorochrome-labeled probes are detected using enzyme-labeled antibodies and visualized by chromogenic enzymatic reactions. The HER2 status (amplified/not amplified and HER2 ratios) was evaluated by the CISH method and compared with results obtained by the FDA-approved FISH method. Of the 72 successfully investigated invasive breast carcinomas, both FISH and CISH detected HER2 amplification in 24 cases and nonamplification was detected in 47 cases. One case showed a discrepancy between FISH and CISH. The concordance between CISH and FISH was found to be almost perfect (98.6%). The correlation between the HER2 ratios obtained by the 2 methods showed excellent correlation (correlation coefficient 0.95). In conclusion, it is possible by dual-color CISH method to demonstrate HER2 genes and chromosome 17 genes, in the same tissue section and reliably assess HER2 status. The CISH method is a very promising alternative to the FISH method.

  3. Using selective chromogenic plates to optimize isolation of group B Streptococcus in pregnant women

    Directory of Open Access Journals (Sweden)

    Romano Mattei

    2014-03-01

    Full Text Available Group B Streptococcus (GBS remains the leading cause of severe bacterial infections (sepsis, meningitis, pneumonia in neonates. We compared the detection of GBS from recto-vaginal swabs on blood agar and two chromogenic media and evaluated their antibiotic susceptibility. A total of 1351 swabs were taken from pregnant women at 35-37 weeks of gestation. Following enrichment in Todd Hewitt broth + nalidixic acid and colistin, the samples were plated on Columbia CNA agar (CNA, chromID Strepto B agar (STRB and Granada Agar (GRAN, respectively. GBS were found in 22.4% of recto-vaginal swabs from pregnant women. Sensitivity, specificity, positive and negative predictive values of GBS detection were 88%, 88%, 81% and 96% for CNA, 99%, 97%, 90% and 99% for STRB and 94%, 99%, 98% e 99% for GRAN; Cohen’s k index concordances for CNA, STREB and GRAN were 0.68, 0.92 and 0.96, respectively. All isolates were susceptible to penicillin, whereas resistances of erythromycin and clindamycin were 40% and 42%, respectively. To conclude, selective broth enrichment combined with chromogenic plates is recommended for GBS screening in pregnant women.

  4. Chromogenic culture media or rapid immunochromatographic test: Which is better for detecting Klebsiella pneumoniae that produce OXA-48 and can they be used in blood and urine specimens.

    Science.gov (United States)

    Genc, Ozlem; Aksu, Evrim

    2018-05-01

    Our goal was to compare a rapid test (OXA-48K-SeT) and four different chromogenic media (CHROMagar KPC, CHROMagar mSuperCARBA, ChromID Carba and ChromID OXA-48) for the detection of OXA-48 producing Klebsiella pneumoniae isolates and spiked urine/blood samples with these bacteria. In total 100 K.pneumoniae isolates, including 60 OXA-48 positive, 15 other carbapenemase producing, 15 Extended spectrum betalactamases (ESBL) positive and 10 carbapenem sensitive K.pneumoniae were included in the study. After all samples were inoculated into all chromogenic media, temocillin discs were placed onto the media. OXA-48K-SeT was studied according to the manufacturer's instructions and the lower detection limit was determined. Sensitivities and specificities of all chromogenic media and rapid test were detected as 100%. All of the OXA-48 producers were found resistant to temocillin on all chromogenic media. The lower detection limit of the rapid assay was determined as 10 6 in both direct bacterial samples and in spiked urine/blood samples. As a result, four chromogenic culture media and OXA-48 K-SeT can be used safely for detection of OXA-48 positive K.pneumoniae isolates. Although direct clinical specimens were not used, our study suggests that this media and OXA-48 K-SeT may be used in patient samples like blood and urine. Further studies are needed to assess this suggestion. Copyright © 2018 Elsevier B.V. All rights reserved.

  5. Qualification of a select one-stage activated partial thromboplastin time-based clotting assay and two chromogenic assays for the post-administration monitoring of nonacog beta pegol.

    Science.gov (United States)

    Tiefenbacher, S; Bohra, R; Amiral, J; Bowyer, A; Kitchen, S; Lochu, A; Rosén, S; Ezban, M

    2017-10-01

    Essentials Nonacog beta pegol (N9-GP) is an extended half-life, recombinant human factor IX (FIX). One-stage clotting (OSC) and chromogenic FIX activity assays were assessed for N9-GP recovery. OSC STA ® -Cephascreen ® , ROX FIX and BIOPHEN FIX chromogenic assays were qualified for N9-GP. Other extended half-life factor products should be assessed in a similar way prior to approval. Background Nonacog beta pegol (N9-GP) is an extended half-life, glycoPEGylated recombinant human factor IX that is under development for the prophylaxis and treatment of bleeding episodes in hemophilia B patients. Considerable reagent-dependent variability has been observed when one-stage clotting assays are used to measure the recovery of recombinant FIX products, including N9-GP. Objective To qualify select one-stage clotting and chromogenic FIX activity assays for measuring N9-GP recovery. Methods The accuracy and precision of the one-stage clotting assay (with the STA-Cephascreen activated partial thromboplastin [APTT] reagent) and the ROX Factor IX and BIOPHEN Factor IX chromogenic assays for measuring N9-GP recovery were assessed in N9-GP-spiked hemophilia B plasma samples in a systematic manner at three independent sites, with manufacturer-recommended protocols and/or site-specific assay setups, including different instruments. Results For each of the three FIX activity assays qualified on five different reagent-instrument systems, acceptable intra-assay and interassay accuracy and precision, dilution integrity, reagent robustness and freeze-thaw and short-term sample stabilities were demonstrated. The STA-Cephascreen assay showed a limited reportable range at one of the three qualification sites, and the BIOPHEN Factor IX assay showed suspect low-end sensitivity at one of the three qualification sites. An individual laboratory would account for these limitations by adjusting the assay's reportable range; thus, these findings are not considered to impact the respective

  6. Analytical assays based on chromogenic and fluorogenic chemosensors for the detection of cyanide

    Directory of Open Access Journals (Sweden)

    Vanderléia Gava Marini

    2010-06-01

    Full Text Available Cyanide (CN– is an anion well–known for its toxicity, being a chemical agent often related to cases of homicide and suicide. Despite being responsible for the toxicity of many animals and plants, it is used in several industrial activities, with innumerous implications in terms of the environment. Due to its high toxicity, the maximum level of CN– concentration allowed by the World Health Organization in potable water is 1.7 µmol/L. This low concentration limit requires methods of visual detection and quantitative determination which are ever more sensitive, simple, reliable, and economical. Advancements in the field of chromogenic and fluorogenic chemosensors for anionic analytes have led to the development of several methodologies for the detection of CN–. Therefore, this review aims to present the main strategies that have been used in the study of quantitative and naked–eye detection of CN– by means of chromogenic and fluorogenic chemosensors. Aspects related to CN–, such as its reactivity, toxicity, applications, and implications in different domains of knowledge, are presented. Recent work involving the development of chemosensors for CN– based on acid–base reactions, chemodosimeters, chromoreactands, and competition assays is also described. In addition, recent studies that make use of nanotechnology to develop strategies for the detection of CN– are also discussed, as well as the prospects envisioned in this field.

  7. Biological activity analysis of native and recombinant streptokinase using clot lysis and chromogenic substrate assay.

    Science.gov (United States)

    Mahboubi, Arash; Sadjady, Seyyed Kazem; Mirzaei Saleh Abadi, Mohammad; Azadi, Saeed; Solaimanian, Roya

    2012-01-01

    DETERMINATION OF STREPTOKINASE ACTIVITY IS USUALLY ACCOMPLISHED THROUGH TWO ASSAY METHODS: a) Clot lysis, b) Chromogenic substrate assay. In this study the biological activity of two streptokinase products, namely Streptase®, which is a native product and Heberkinasa®, which is a recombinant product, was determined against the third international reference standard using the two forementioned assay methods. The results indicated that whilst the activity of Streptase® was found to be 101 ± 4% and 97 ± 5% of the label claim with Clot lysis and Chromogenic substrate assay respectively, for Heberkinasa® the potency values obtained were 42 ± 5% and 92.5 ± 2% of the label claim respectively. To shed some light on the reason for this finding, the n-terminal sequence of the streptokinase molecules present in the two products was determined. The results showed slight differences in the amino acid sequence of the recombinant product in comparison to the native one at the amino terminus. This finding supports those of other workers who found that n-terminal sequence of the streptokinase molecule can have significant effect on the activity of this protein.

  8. Changes in the chromogene properties of the betalaine

    International Nuclear Information System (INIS)

    Rodriguez N, S.; Pinedo S, A.; Amador V, P.; Chacon R, A.; Arcos P, A.; Vega C, H.R.

    2005-01-01

    The changes of coloration of four natural extracts were determined in function of the absorbed dose taken place by a source of rays gamma of Cs-137. The used natural extracts contain betalaine that are natural pigments of some plants as the beet root, of there their name. They are also in abundant form in the fruits (tunas) of some species of the Opuntia gender. The extracts were obtained by maceration, starting from beet root and three tuna varieties, and they were stabilized to pH 5.5 the change of coloration you determines in a spectrophotometer ultraviolet/visible by means of the absorbance from the samples to photons of 475, 535 and 600 nm of wave longitude. The absorbance was measured, to different intervals of time. The relationship settled down between the absorbed dose and the chromogene properties of the pigment, with the intention of using it as possible dosemeter. (Author)

  9. Putting the colours into chromogenic in situ hybridization (CISH).

    Science.gov (United States)

    Shipley, J

    2006-09-01

    Recurrent genomic alterations are the hallmarks of particular cancers. Application of molecular cytogenetic technologies to tumour material in order to detect these alterations has become important for molecular diagnostics and research. A dual-colour chromogenic in situ hybridization (dc-CISH) method described recently in the Journal of Pathology has the advantage of visualizing two probes simultaneously with the ability to discern morphological features. In addition, the bright field microscopy required is readily accessible to many laboratories. The approach has been validated by comparison of results with standard analyses for HER-2 amplification status in formalin-fixed, paraffin-embedded breast cancers and is applicable to the analysis of other clinically relevant genomic aberrations as well as of use in research investigations. Copyright (c) 2006 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

  10. Measuring factor IX activity of nonacog beta pegol with commercially available one-stage clotting and chromogenic assay kits: a two-center study.

    Science.gov (United States)

    Bowyer, A E; Hillarp, A; Ezban, M; Persson, P; Kitchen, S

    2016-07-01

    Essentials Validated assays are required to precisely measure factor IX (FIX) activity in FIX products. N9-GP and two other FIX products were assessed in various coagulation assay systems at two sites. Large variations in FIX activity measurements were observed for N9-GP using some assays. One-stage and chromogenic assays accurately measuring FIX activity for N9-GP were identified. Background Measurement of factor IX activity (FIX:C) with activated partial thromboplastin time-based one-stage clotting assays is associated with a large degree of interlaboratory variation in samples containing glycoPEGylated recombinant FIX (rFIX), i.e. nonacog beta pegol (N9-GP). Validation and qualification of specific assays and conditions are necessary for the accurate assessment of FIX:C in samples containing N9-GP. Objectives To assess the accuracy of various one-stage clotting and chromogenic assays for measuring FIX:C in samples containing N9-GP as compared with samples containing rFIX or plasma-derived FIX (pdFIX) across two laboratory sites. Methods FIX:C, in severe hemophilia B plasma spiked with a range of concentrations (from very low, i.e. 0.03 IU mL(-1) , to high, i.e. 0.90 IU mL(-1) ) of N9-GP, rFIX (BeneFIX), and pdFIX (Mononine), was determined at two laboratory sites with 10 commercially available one-stage clotting assays and two chromogenic FIX:C assays. Assays were performed with a plasma calibrator and different analyzers. Results A high degree of variation in FIX:C measurement was observed for one-stage clotting assays for N9-GP as compared with rFIX or pdFIX. Acceptable N9-GP recovery was observed in the low-concentration to high-concentration samples tested with one-stage clotting assays using SynthAFax or DG Synth, or with chromogenic FIX:C assays. Similar patterns of FIX:C measurement were observed at both laboratory sites, with minor differences probably being attributable to the use of different analyzers. Conclusions These results suggest that, of the

  11. A protocol describing the use of a recombinant protein-based, animal product-free medium (APEL) for human embryonic stem cell differentiation as spin embryoid bodies.

    Science.gov (United States)

    Ng, Elizabeth S; Davis, Richard; Stanley, Edouard G; Elefanty, Andrew G

    2008-01-01

    In order to promote the uniform and reproducible differentiation of human embryonic stem cells (HESCs) in response to exogenously added growth factors, we have developed a method (spin embryoid bodies (EBs)) that uses a recombinant protein-based, animal product-free medium in which HESCs are aggregated by centrifugation to form EBs. In this protocol we describe the formulation of this medium, denoted APEL (Albumin Polyvinylalcohol Essential Lipids), and its use in spin EB differentiation of HESCs. We also describe a more economical variant, BPEL (Bovine Serum Albumin (BSA) Polyvinylalchohol Essential Lipids), in which BSA replaces the recombinant human albumin. The integration of a medium that includes only defined and recombinant components with a defined number of cells to initiate EB formation results in a generally applicable, robust platform for growth factor-directed HESC differentiation.

  12. High concentration of human lactoferrin in milk of rhLf-transgenic cows relieves signs of bovine experimental Staphylococcus chromogenes intramammary infection.

    Science.gov (United States)

    Simojoki, Heli; Hyvönen, Paula; Orro, Toomas; Pyörälä, Satu

    2010-08-15

    Six transgenic cows producing recombinant human lactoferrin (rhLf) in their milk and five normal cows at the same lactation stage were experimentally infected with Staphylococcus chromogenes to study the effect of a high concentration of lactoferrin in milk. Coagulase-negative staphylococci such as S. chromogenes have become very common as agents causing mild or subclinical mastitis. All transgenic cows became infected but showed no clinical signs, unlike the control cows, which developed mild clinical mastitis. Transgenic cows eliminated bacteria faster from the quarters than did the controls. Local clinical signs were milder, and the inflammatory reaction assessed by NAGase activity in the milk and by the concentration of milk amyloid A was lower in the transgenic cows. The mild response probably reflected the rapid elimination of bacteria. The milk concentration of rhLf remained constant throughout the study period, but the total concentration of bovine lactoferrin in the milk peaked in both groups at 46h post-challenge. Three cows, all in the control group, exhibited systemic acute phase response as increased concentrations of serum amyloid A in the blood circulation. Transgenic cows with a high concentration of human lactoferrin in their milk seemed to be protected from clinical disease and from prolonged inflammatory reaction, but not from experimental intramammary infection induced by S. chromogenes. Copyright 2010 Elsevier B.V. All rights reserved.

  13. Kinetics of intravenous radiographic contrast medium injections as used on CT: simulation with time delay differential equations in a basic human cardiovascular multicompartment model.

    Science.gov (United States)

    Violon, D

    2012-12-01

    To develop a multicompartment model of only essential human body components that predicts the contrast medium concentration vs time curve in a chosen compartment after an intravenous injection. Also to show that the model can be used to time adequately contrast-enhanced CT series. A system of linked time delay instead of ordinary differential equations described the model and was solved with a Matlab program (Matlab v. 6.5; The Mathworks, Inc., Natick, MA). All the injection and physiological parameters were modified to cope with normal or pathological situations. In vivo time-concentration curves from the literature were recalculated to validate the model. The recalculated contrast medium time-concentration curves and parameters are given. The results of the statistical analysis of the study findings are expressed as the median prediction error and the median absolute prediction error values for both the time delay and ordinary differential equation systems; these are situated well below the generally accepted maximum 20% limit. The presented program correctly predicts the time-concentration curve of an intravenous contrast medium injection and, consequently, allows an individually tailored approach of CT examinations with optimised use of the injected contrast medium volume, as long as time delay instead of ordinary differential equations are used. The presented program offers good preliminary knowledge of the time-contrast medium concentration curve after any intravenous injection, allowing adequate timing of a CT examination, required by the short scan time of present-day scanners. The injected volume of contrast medium can be tailored to the individual patient with no more contrast medium than is strictly needed.

  14. Anionic chromogenic chemosensors highly selective for fluoride or cyanide based on 4-(4-Nitrobenzylideneamine)phenol

    OpenAIRE

    Nicoleti,Celso R.; Marini,Vanderléia G.; Zimmermann,Lizandra M.; Machado,Vanderlei G.

    2012-01-01

    4-(4-Nitrobenzylideneamine)phenol was used in two strategies allowing the highly selective detection of F- and CN-. Firstly, the compound in acetonitrile acts as a chromogenic chemosensor based on the idea that more basic anions cause its deprotonation (colorless solution), generating a colored solution containing phenolate. The discrimination of CN- over F- was obtained by adding 1.4% water to acetonitrile: water preferentially solvates F-, leaving the CN- free to deprotonate the compound. A...

  15. Zn(II)-cyclam based chromogenic sensors for recognition of ATP in aqueous solution under physiological conditions and their application as viable staining agents for microorganism.

    Science.gov (United States)

    Mahato, Prasenjit; Ghosh, Amrita; Mishra, Sanjiv K; Shrivastav, Anupama; Mishra, Sandhya; Das, Amitava

    2011-05-02

    Two chromogenic complexes, L.Zn (where L is (E)-4-((4-(1,4,8,11-tetraazacyclotetradecan-1-ylsulfonyl)phenyl)diazenyl)-N,N-dimethylaniline) and its [2]pseudorotaxane form (α-CD.L.Zn), were found to bind preferentially to adenosine triphosphate (ATP), among all other common anions and biologically important phosphate (AMP, ADP, pyrophosphate, and phosphate) ions in aqueous HEPES buffer medium of pH 7.2. Studies with live cell cultures of prokaryotic microbes revealed that binding of these two reagents to intercellular ATP, produced in situ, could be used in delineating the gram-positive and the gram-negative bacteria. More importantly, these dyes were found to be nontoxic to living microbes (eukaryotes and prokaryotes) and could be used for studying the cell growth dynamics. Binding to these two viable staining agents to intercellular ATP was also confirmed by spectroscopic studies on cell growth in the presence of different respiratory inhibitors that influence the intercellular ATP generation. © 2011 American Chemical Society

  16. Chromogenic platform based on recombinant Drosophila melanogaster acetylcholinesterase for visible unidirectional assay of organophosphate and carbamate insecticide residues

    Energy Technology Data Exchange (ETDEWEB)

    Han Zheng [Institute for Agri-food Standards and Testing Technology, Shanghai Academy of Agricultural Sciences, 1018 Jinqi Road, Shanghai 201403 (China); Chi Chensen [School of Life Science and Biotechnology, Bor Luh Food Safety Center, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai 200240 (China); Bai Bing; Liu Gang; Rao Qinxiong [Institute for Agri-food Standards and Testing Technology, Shanghai Academy of Agricultural Sciences, 1018 Jinqi Road, Shanghai 201403 (China); Peng Shaojie [Institute of Shanghai Food and Drug Supervision, 615 Liuzhou Road, Shanghai 200233 (China); Liu Hong [Shanghai Municipal Center for Disease Control and Prevention, 1380 Zhongshan West Road, Shanghai 200336 (China); Zhao Zhihui [Institute for Agri-food Standards and Testing Technology, Shanghai Academy of Agricultural Sciences, 1018 Jinqi Road, Shanghai 201403 (China); Zhang Dabing [School of Life Science and Biotechnology, Bor Luh Food Safety Center, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai 200240 (China); Wu Aibo, E-mail: wuaibo@saas.sh.cn [Institute for Agri-food Standards and Testing Technology, Shanghai Academy of Agricultural Sciences, 1018 Jinqi Road, Shanghai 201403 (China)

    2012-03-30

    Highlight: Black-Right-Pointing-Pointer A visible chromogenic platform for rapid analysis of OP and CM insecticide residues was developed. Black-Right-Pointing-Pointer The assay has the capabilities of both qualitative measurement and quantitative analysis. Black-Right-Pointing-Pointer The sensitivity, capabilities of resisting interferences and storage stability were desirable. Black-Right-Pointing-Pointer Matrix effects were acceptable and detection performance was satisfactory in real application. - Abstract: In this study we propose a chromogenic platform for rapid analysis of organophosphate (OP) and carbamate (CM) insecticide residues, based on recombinant Drosophila melanogaster acetylcholinesterase (R-DmAChE) as enzyme and indoxyl acetate as substrate. The visible chromogenic strip had the advantages identical to those of commonly used lateral flow assays (LFAs) with utmost simplicity in sample loading and result observation. After optimization, depending on the color intensity (CI) values, the well-established assay has the capabilities of both qualitative measurement via naked eyes and quantitative analysis by colorimetric reader with the desirable IC{sub 50} values against the tested six insecticides (0.06 {mu}g mL{sup -1} of carbofuran, 0.28 {mu}g mL{sup -1} of methomyl, 0.03 {mu}g mL{sup -1} of dichlorvos, 31.6 {mu}g mL{sup -1} of methamidophos, 2.0 {mu}g mL{sup -1} of monocrotophos, 6.3 {mu}g mL{sup -1} of omethoate). Acceptable matrix effects and satisfactory detection performance were confirmed by in-parallel LC-MS/MS analysis in different vegetable varieties at various spiked levels of 10{sup -3} to 10{sup 1} {mu}g g{sup -1}. Overall, the testified suitability and applicability of this novel platform meet the requirements for practical use in food safety management and environmental monitoring, especially in the developing world.

  17. Chromogenic platform based on recombinant Drosophila melanogaster acetylcholinesterase for visible unidirectional assay of organophosphate and carbamate insecticide residues

    International Nuclear Information System (INIS)

    Han Zheng; Chi Chensen; Bai Bing; Liu Gang; Rao Qinxiong; Peng Shaojie; Liu Hong; Zhao Zhihui; Zhang Dabing; Wu Aibo

    2012-01-01

    Highlight: ► A visible chromogenic platform for rapid analysis of OP and CM insecticide residues was developed. ► The assay has the capabilities of both qualitative measurement and quantitative analysis. ► The sensitivity, capabilities of resisting interferences and storage stability were desirable. ► Matrix effects were acceptable and detection performance was satisfactory in real application. - Abstract: In this study we propose a chromogenic platform for rapid analysis of organophosphate (OP) and carbamate (CM) insecticide residues, based on recombinant Drosophila melanogaster acetylcholinesterase (R-DmAChE) as enzyme and indoxyl acetate as substrate. The visible chromogenic strip had the advantages identical to those of commonly used lateral flow assays (LFAs) with utmost simplicity in sample loading and result observation. After optimization, depending on the color intensity (CI) values, the well-established assay has the capabilities of both qualitative measurement via naked eyes and quantitative analysis by colorimetric reader with the desirable IC 50 values against the tested six insecticides (0.06 μg mL −1 of carbofuran, 0.28 μg mL −1 of methomyl, 0.03 μg mL −1 of dichlorvos, 31.6 μg mL −1 of methamidophos, 2.0 μg mL −1 of monocrotophos, 6.3 μg mL −1 of omethoate). Acceptable matrix effects and satisfactory detection performance were confirmed by in-parallel LC–MS/MS analysis in different vegetable varieties at various spiked levels of 10 −3 to 10 1 μg g −1 . Overall, the testified suitability and applicability of this novel platform meet the requirements for practical use in food safety management and environmental monitoring, especially in the developing world.

  18. Chromogene properties of the betalains before gamma photons

    International Nuclear Information System (INIS)

    Rodriguez N, S.; Quintero M, C. L.; Vega C, H. R.

    2010-09-01

    The coloration changes of four natural extracts in function of the absorbed dose produced by a gamma rays source of 137 Cs have been studied. The natural extracts were obtained of tuna varieties that contain betalains that are natural pigments of some plants as the beet of where is derived its name. These also are found in abundant form in the fruits of some species of opuntia genus (tunas). The extracts were obtained by maceration, starting from beet and three tuna varieties that were stabilized to a p H of 5.5. The extracts were exposed to the gamma rays of a 137 Cs source and the change in the coloration was observed by means of an ultra violet/visible spectrophotometer through of the absorption of the samples to photons of wave longitude 535 nm. The absorption was measured, to different time intervals. The relation between the absorbed dose in D w water and the chromogene properties of the pigment was established, with the intention of using it as possible dosemeter. (Author)

  19. Comparison of Fluorescence In Situ Hybridization and Chromogenic In Situ Hybridization for Low and High Throughput HER2 Genetic Testing

    DEFF Research Database (Denmark)

    Poulsen, Tim S; Espersen, Maiken Lise Marcker; Kofoed, Vibeke

    2013-01-01

    cancer patients with HER2 immunohistochemistry (IHC) results scored as 0/1+, 2+, and 3+. HER2 genetic status was analysed using chromogenic in situ hybridization (CISH) and fluorescence in situ hybridization (FISH). Scoring results were documented through digital image analysis. The cancer region...

  20. CHROMagar COL-APSE: a selective bacterial culture medium for the isolation and differentiation of colistin-resistant Gram-negative pathogens

    DEFF Research Database (Denmark)

    Abdul Momin, Muhd Haziq F; Bean, David C; Hendriksen, Rene S.

    2017-01-01

    medium (SuperPolymyxin). Methodology. The medium was challenged with 84 isolates, including polymyxin B (POL B)-susceptible and -resistant type strains and colistin (COL)-resistant organisms recovered from human and animal samples. Susceptibility to COL and POL B was determined by agar dilution and broth...... microtitre dilution. The lower limit for the detection of COL-resistant organisms was also calculated for both CHROMagar COL-APSE and SuperPolymyxin media. The ability to isolate and correctly differentiate COL-resistant organisms within mixed cultures was also assessed and compared using both media. Results...

  1. Proton quasi-elastic scattering at 600MeV on the. cap alpha. -substructure of medium nuclei. [Differential cross sections

    Energy Technology Data Exchange (ETDEWEB)

    Anne, R [CEA Centre d' Etudes Nucleaires de Saclay, 91 - Gif-sur-Yvette (France). Service d' Entretien et de Developpement des Appareils de Physique; Delpierre, P; Kahane, J; Sene, R [College de France, 75 - Paris. Lab. de Physique Corpusculaire; Devaux, A; Landaud, G [Clermont-Ferrand Univ., 63 (France); Yonnet, J [Caen Univ., 14 (France). Lab. de Physique Corpusculaire

    1975-01-01

    Alpha knock out from light and medium nuclei up to /sup 40/Ca was investigated. Preliminary values of the differential cross sections are given for /sup 6/Li and /sup 12/C nuclei. The p(R) recoil momentum distributions show a maximum at p(R)=0.

  2. In situ analysis of proteins at high temperatures mediated by capillary-flow hydrothermal UV-vis spectrophotometer with a water-soluble chromogenic reagent.

    Science.gov (United States)

    Kawamura, Kunio; Nagayoshi, Hiroki; Yao, Toshio

    2010-05-14

    In situ monitoring of quantities, interactions, and conformations of proteins is essential for the study of biochemistry under hydrothermal environments and the analysis of hyperthermophilic organisms in natural hydrothermal systems on Earth. We have investigated the potential of a capillary-flow hydrothermal UV-vis spectrophotometer (CHUS) for performing in situ measurements of proteins and determining their behavior at extremely high temperatures, in combination with a chromogenic reagents probe, which interacts with the proteins. The spectral shift obtained using a combination of water-soluble porphyrin (TPPS) and bovine serum albumin (BSA) was the best among the spectral shifts obtained using different combinations of chromogenic reagents and proteins. The association behavior of TPPS with BSA was investigated in detail using CHUS at temperatures up to 175 degrees C and the association constant (K(ass)) of TPPS with BSA was successfully determined at temperatures up to 100 degrees C. The lnK(ass) values were inversely proportional to the T(-1) values in the temperature range 50-100 degrees C. These analyses showed for the first time that the decrease of association of TPPS with BSA is due to the conformational change, fragmentation, and/or denaturing of BSA rather than the decrease of the hydrophobic association between TPPS and BSA. This study conclusively demonstrates the usability of the CHUS system with a chromogenic reagent as an in situ detection and measurement system for thermostable proteins at extremely high temperatures. Copyright 2010 Elsevier B.V. All rights reserved.

  3. Evaluation of Myc Gene Amplification in Prostate Cancer Using a Dual Color Chromogenic in-Situ Hybridization (Dual CISH Assay

    Directory of Open Access Journals (Sweden)

    Daniel Lerda

    2013-04-01

    Full Text Available Objetive: The overall purpose of the study was to demonstrate applicability of the Dako dual-color chromogenic in situ hybridization (CISH assay (DAKO Denmark, Glostrup with respect to fluorescence in situ hybridization (FISH probes MYC-C. Methods: MYC gene amplification by FISH and Dako dual-color CISH Results: The study showed that the dual-color CISH assay can convert Texas red and fluorescein isothiocyanate (FITC signals into chromogenic signals. The dual –color CISH assay was performed on 40 cases of prostate cancer. Amplification was identified in 12 of 40 (30% tumors. No amplification was seen in 28 of 40 (70% tumors. FISH data were available in total of 40 tumors. All tumors showed concordant results between dual-color CISH and FISH for classifying a tumor as MYC amplified or not amplified. Conclusions: We conclude that dual-color Dako CISH assay is an accurate method for determining MYC gene amplification with added advantages that make it a more practically useful method. [J Interdiscipl Histopathol 2013; 1(2.000: 81-84

  4. Growth factors and medium hyperglycemia induce Sox9+ ductal cell differentiation into β cells in mice with reversal of diabetes

    Science.gov (United States)

    Zhang, Mingfeng; Lin, Qing; Qi, Tong; Wang, Tiankun; Chen, Ching-Cheng; Riggs, Arthur D.; Zeng, Defu

    2016-01-01

    We previously reported that long-term administration of a low dose of gastrin and epidermal growth factor (GE) augments β-cell neogenesis in late-stage diabetic autoimmune mice after eliminating insulitis by induction of mixed chimerism. However, the source of β-cell neogenesis is still unknown. SRY (sex-determining region Y)-box 9+ (Sox9+) ductal cells in the adult pancreas are clonogenic and can give rise to insulin-producing β cells in an in vitro culture. Whether Sox9+ ductal cells in the adult pancreas can give rise to β cells in vivo remains controversial. Here, using lineage-tracing with genetic labeling of Insulin- or Sox9-expressing cells, we show that hyperglycemia (>300 mg/dL) is required for inducing Sox9+ ductal cell differentiation into insulin-producing β cells, and medium hyperglycemia (300–450 mg/dL) in combination with long-term administration of low-dose GE synergistically augments differentiation and is associated with normalization of blood glucose in nonautoimmune diabetic C57BL/6 mice. Short-term administration of high-dose GE cannot augment differentiation, although it can augment preexisting β-cell replication. These results indicate that medium hyperglycemia combined with long-term administration of low-dose GE represents one way to induce Sox9+ ductal cell differentiation into β cells in adult mice. PMID:26733677

  5. Comparison of selective agars recommended by method ISO 11290-1 and chromogenic agars for the isolation of Listeria sp. in refrigerated sausages

    Directory of Open Access Journals (Sweden)

    Thalyta Marina Benetti

    2012-12-01

    Full Text Available The aim of this study was to determine the prevalence of Listeria sp. in refrigerated sausages, and to compare the performance of the selective plating media employed in the ISO 11290-1 method (PALCAM and Oxford agars with chromogenic agars (Chromogenic Listeria agars CM 1080 (OCLA and CM 1084. The prevalence of Listeria sp. detected was 52.9%, comprising 13.7% L. monocytogenes strains. The efficacy of the four agars for the isolation of L. monocytogenes proved to be satisfactory. Despite differences in composition of the chromogenic media assessed, these disparities did not affect concordance among results. However, PALCAM agar was shown to suppress other microorganisms more effectively, being more applicable for detecting Listeria strains present in lower quantities. Based on these results, the use of PALCAM agar, in combination with a chromogenic media, is recommended for enhanced isolation of atypical Listeria sp. strains in meat products.Este estudo teve como objetivo a análise da prevalência de Listeria sp. em linguiças resfriadas e a comparação dos meios seletivos utilizados no plaqueamento do método ISO 11290-1 (Ágar PALCAM e Ágar Oxford, e ágares cromogênicos (Ágares Listeria Cromogênico CM 1080 (OCLA e CM 1084 (ISO. A frequência de Listeria sp. foi de 52,9%, sendo que destas, 13,7% corresponderam à L. monocytogenes. A eficácia dos quatro ágares para o isolamento de L. monocytogenes demonstrou-se satisfatória. Apesar de haver algumas diferenças nas composições dos meios cromogênicos analisados, estas não pareceram influenciar nas concordâncias entre os resultados expressos. Contudo, o ágar PALCAM mostrou-se mais eficaz na supressão de outros micro-organismos, aumentando, assim, a possibilidade de detecção de espécies de Listeria presentes em número reduzido. Através deste trabalho sugere-se a utilização do ágar PALCAM associado a um meio cromogênico para aumentar a chance de isolamento de cepas at

  6. Spectrophotometric determination of copper in alkaline solutions and evaluation of some hydroxy-substituted 1,10-phenanthrolines as chromogenic reagents.

    Science.gov (United States)

    Dunbar, W E; Schilt, A A

    1972-09-01

    Seven new hydroxy-substituted 1,10-phenanthroline derivatives have been evaluated as chromogenic reagents for the determination of copper in strongly alkaline solution. The most sensitive of these, 2,9-dimethyl-4,7-dihydroxy-1,10-phenanthroline, has proven to be highly effective in a simple, rapid procedure for determining trace amounts of copper in sodium hydroxide, potassium carbonate, sodium phosphate or ammonium hydroxide.

  7. Improved Screening Method for the Selection of Wine Yeasts Based on Their Pigment Adsorption Activity

    Directory of Open Access Journals (Sweden)

    Andrea Caridi

    2013-01-01

    Full Text Available The aim of this research is to improve an existing low-cost and simple but consistent culturing technique for measuring the adsorption of grape skin pigments on yeasts, comprising: (i growing yeasts in Petri dishes on chromogenic grape-skin-based medium, (ii photographing the yeast biomass, (iii measuring its red, green, and blue colour components, and (iv performing the statistical analysis of the data. Twenty strains of Saccharomyces cerevisiae were grown on different lots of the chromogenic medium, prepared using grape skins from dark cultivars Greco Nero, Magliocco and Nero d’Avola. Microscale wine fermentation trials were also performed. Wide and significant differences among wine yeasts were observed. The chromogenic grape-skin-based medium can be prepared using any grape cultivar, thus allowing the specific selection of the most suitable strain of Saccharomyces cerevisiae for each grape must, mainly for red winemaking. The research provides a useful tool to characterize wine yeasts in relation to pigment adsorption, allowing the improvement of wine colour.

  8. Patients with deep venous thrombosis and thrombophilia risk factors have a specific prolongation of the lag time in a chromogenic thrombin generation assay

    NARCIS (Netherlands)

    Haas, F.J.L.M.; Kluft, C.; Biesma, D.H.; Schutgens, R.E.G.

    2011-01-01

    The objective of the present study was to evaluate the influence of thrombophilia risk factors on variables of a chromogenic thrombin generation assay (ETP) in a setting with acute deep venous thrombosis (DVT) and non-DVT patients. In 152 outpatients suspected for DVT, the results of thrombophilia

  9. Improved Survival and Initiation of Differentiation of Human Induced Pluripotent Stem Cells to Hepatocyte-Like Cells upon Culture in William's E Medium followed by Hepatocyte Differentiation Inducer Treatment.

    Directory of Open Access Journals (Sweden)

    Minoru Tomizawa

    Full Text Available Hepatocyte differentiation inducer (HDI lacks both glucose and arginine, but is supplemented with galactose and ornithine, and is added together with other reagents such as apoptosis inhibitor and oncostatin M. Although human induced pluripotent stem (iPS cells initiate hepatocyte differentiation, most die within 7 days. In this study, we investigated both HDI and conventional media for their potential to improve cell survival.201B7 iPS cells were cultured in conventional media. This consisted of three cycles of 5-day culture in William's E (WE medium, followed by a 2-day culture in HDI.Expression levels of α-feto protein (AFP were higher in cells cultured in WE and in Dulbecco's Modified Eagle's Medium/Nutrient F-12 Ham (DF12. 201B7 cells expressed the highest AFP and albumin (ALB when cultured in HDI for 2 days following 7-day culture in WE. After three cycles of 5-day culture in WE followed by 2 days in HDI, 201B7 cells expressed AFP and ALB 54 ± 2.3 (average ± standard deviation and 73 ± 15.1 times higher, respectively, than those cultured in ReproFF (feeder-free condition.201B7 cells survived culture in WE for 7 days followed HDI for 2 days. After three cycles of culture under these conditions, hepatocyte differentiation was enhanced, as evidenced by increased AFP and ALB expression.

  10. Chromogenic in situ hybridisation for the assessment of HER2 status in breast cancer: an international validation ring study

    OpenAIRE

    van de Vijver, Marc; Bilous, Michael; Hanna, Wedad; Hofmann, Manfred; Kristel, Petra; Penault-Llorca, Frédérique; Rüschoff, Josef

    2007-01-01

    Introduction Before any new methodology can be introduced into the routine diagnostic setting it must be technically validated against the established standards. To this end, a ring study involving five international pathology laboratories was initiated to validate chromogenic in situ hybridisation (CISH) against fluorescence in situ hybridisation (FISH) and immunohistochemistry (IHC) as a test for assessing human epidermal growth factor receptor 2 (HER2) status in breast cancer. Methods Each...

  11. In vitro cementoblast-like differentiation of postmigratory neural crest-derived p75+ stem cells with dental follicle cell conditioned medium

    International Nuclear Information System (INIS)

    Wen, Xiujie; Liu, Luchuan; Deng, Manjing; Liu, Rui; Zhang, Li; Nie, Xin

    2015-01-01

    Cranial neural crest-derived cells (CNCCs) play important role in epithelial–mesenchymal interactions during tooth morphogenesis. However, the heterogeneity of CNCCs and their tendency to spontaneously differentiate along smooth muscle or osteoblast lineages in vitro limit further understanding of their biological properties. We studied the differentiation properties of isolated rat embryonic postmigratory CNCCs, expressing p75 neurotrophin receptor (p75NTR). These p75NTR positive (p75 + ) CNCCs, isolated using fluorescence activated cell sorter, exhibited fibroblast-like morphology and characteristics of mesenchymal stem cells. Incubation of p75 + CNCCs in dental follicle cell conditioned medium (DFCCM) combined with dentin non-collagenous proteins (dNCPs), altered their morphological features to cementoblast-like appearance. These cells also showed low proliferative activity, high ALP activity and significantly increased calcified nodule formation. Markers related to mineralization or specific to cementoblast lineage were highly expressed in dNCPs/DFCCM-treated p75 + cells, suggesting their differentiation along cementoblast-like lineage. p75 + stem cells selected from postmigratory CNCCs represent a pure stem cell population and could be used as a stem cell model for in vitro studies due to their intrinsic ability to differentiate to neuronal cells and transform from neuroectoderm to ectomesenchyme. They can provide a potential stem cell resource for tooth engineering studies and help to further investigate mechanisms of epithelial–mesenchymal interactions in tooth morphogenesis. - Highlights: • Cranial neural crest-derived cells (CNCCs) take part in tooth morphogenesis. • positive (p75 + ) CNCCs are fibroblast-like and resemble mesenchymal stem cells. • p75 + CNCCs in dental follicle cell medium (DFCCM/dNCP) appear like cementoblasts. • DFCCM/dNCP-treated p75 + cells express cementoblast specific mineralization markers. • p75 + cells are pure stem

  12. Factors responsible for subclinical mastitis in cows caused by Staphylococcus chromogenes and its susceptibility to antibiotics based on bap, fnbA, eno, mecA, tetK, and ermA genes.

    Science.gov (United States)

    Bochniarz, M; Adaszek, Ł; Dzięgiel, B; Nowaczek, A; Wawron, W; Dąbrowski, R; Szczubiał, M; Winiarczyk, S

    2016-12-01

    The aim of this study was to recognize selected factors of virulence determining the adhesion of Staphylococcus chromogenes to cows' udder tissues in subclinical mastitis and to evaluate the susceptibility of this pathogen to antibiotics. The subjects of the study were 38 isolates of Staph. chromogenes from 335 samples of milk from cows with subclinical coagulase-negative staphylococci mastitis. Somatic cell count ranged between 216,000 and 568,000/mL of milk (average 356,000/mL of milk). We confirmed the ability to produce slime in 24 isolates (63.2%), and the ability to produce protease in 29 isolates (76.3%). In each slime-producing isolate, the bap gene was not found, and the fnbA and eno genes were not detected. In vitro tests showed that ceftiofur had the highest effectiveness against Staph. chromogenes (89.5% of susceptible isolates). Minimum inhibitory concentrations ranged from 0.06 to 2µg/mL for susceptible isolates. The minimum concentrations required to inhibit growth of 90 and 50% of the isolates for ceftiofur were at or below the cutoffs recommended by the Clinical and Laboratory Standards Institute (2 and 0.06µg/mL, respectively). A significant percentage of the isolates were susceptible to other β-lactam antibiotics: amoxicillin with clavulanic acid (84.2%) and ampicillin (81.6%). The lowest effectiveness among β-lactams was for penicillin (73.7% of susceptible isolates), and the minimum inhibitory concentration for penicillin ranged from chromogenes (71.1 and 63.2% of susceptible isolates, respectively). The genes tetK (6 isolates) and ermA (1 isolate) were also detected. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  13. Bone Marrow Mesenchymal Stem Cells Enhance the Differentiation of Human Switched Memory B Lymphocytes into Plasma Cells in Serum-Free Medium

    Directory of Open Access Journals (Sweden)

    Guillaume Bonnaure

    2016-01-01

    Full Text Available The differentiation of human B lymphocytes into plasma cells is one of the most stirring questions with regard to adaptive immunity. However, the terminal differentiation and survival of plasma cells are still topics with much to be discovered, especially when targeting switched memory B lymphocytes. Plasma cells can migrate to the bone marrow in response to a CXCL12 gradient and survive for several years while secreting antibodies. In this study, we aimed to get closer to niches favoring plasma cell survival. We tested low oxygen concentrations and coculture with mesenchymal stem cells (MSC from human bone marrow. Besides, all cultures were performed using an animal protein-free medium. Overall, our model enables the generation of high proportions of CD38+CD138+CD31+ plasma cells (≥50% when CD40-activated switched memory B lymphocytes were cultured in direct contact with mesenchymal stem cells. In these cultures, the secretion of CXCL12 and TGF-β, usually found in the bone marrow, was linked to the presence of MSC. The level of oxygen appeared less impactful than the contact with MSC. This study shows for the first time that expanded switched memory B lymphocytes can be differentiated into plasma cells using exclusively a serum-free medium.

  14. Osteogenic medium is superior to growth factors in differentiation of human adipose stem cells towards bone-forming cells in 3D culture

    Directory of Open Access Journals (Sweden)

    L Tirkkonen

    2013-01-01

    Full Text Available Human adipose stem cells (hASCs have been recently used to treat bone defects in clinical practice. Yet there is a need for more optimal scaffolds and cost-effective approaches to induce osteogenic differentiation of hASCs. Therefore, we compared the efficiency of bone morphogenetic proteins (BMP-2 and BMP-7, vascular endothelial growth factor (VEGF, and osteogenic medium (OM for the osteo-induction of hASCs in 3D culture. In addition, growth factors were tested in combination with OM. Commercially available bioactive glass scaffolds (BioRestore and biphasic calcium phosphate granules (BoneCeramic were evaluated as prospective carriers for hASCs. Both biomaterials supported hASC-viability, but BioRestore resulted in higher cell number than BoneCeramic, whereas BoneCeramic supported more significant collagen production. The most efficient osteo-induction was achieved with plain OM, promoting higher alkaline phosphatase activity and collagen production than growth factors. In fact, treatment with BMP-2 or VEGF did not increase osteogenic differentiation or cell number significantly more than maintenance medium with either biomaterial. Moreover, BMP-7 treatment consistently inhibited proliferation and osteogenic differentiation of hASCs. Interestingly, there was no benefit from growth factors added to OM. This is the first study to demonstrate that OM enhances hASC-differentiation towards bone-forming cells significantly more than growth factors in 3D culture.

  15. Dithizone as novel and efficient chromogenic probe for cyanide detection in aqueous media through nucleophilic addition into diazenylthione moiety.

    Science.gov (United States)

    Tavallali, Hossein; Deilamy-Rad, Gohar; Parhami, Abolfath; Kiyani, Sajede

    2014-01-01

    A new selective chemodosimeter probe was developed by the introduction of dithizone (DTZ) as a simple and available dye for detection of cyanide in aqueous media which enables recognition of cyanide over other competing anions such as acetate, dihydrogen phosphate, fluoride and benzoate through covalent bonding. The sensing properties of DTZ were investigated in DMSO/H2O (1:9) and have demonstrated a very high selectivity toward the cyanide anions. A reasonable recognition mechanism was suggested using UV-Vis, (1)H NMR and FTIR spectroscopy techniques. Time dependent density function theory (TDDFT) computations of UV-Vis excitation for DTZ2-CN adduct agreed well with our experimental findings. The detection limit of the new chromogenic probe was measured to be 0.48 μmol L(-1) which is much lower than most recently reported chromogenic probes for cyanide determination. The analytical utility of the method for the analysis of cyanide ions in electroplating wastewater (EPWW), human serum, tap and mineral water samples was demonstrated and the results were compared successfully with the conventional reference method. The short time response and the detection by the naked eye make the method available for the detection and quantitative determination of cyanide in a variety of real samples. Copyright © 2013 Elsevier B.V. All rights reserved.

  16. Defined culture medium for stem cell differentiation: applicability of serum-free conditions in the mouse embryonic stem cell test.

    Science.gov (United States)

    Riebeling, Christian; Schlechter, Katharina; Buesen, Roland; Spielmann, Horst; Luch, Andreas; Seiler, Andrea

    2011-06-01

    The embryonic stem cell test (EST) is a validated method to assess the developmental toxicity potency of chemicals. It was developed to reduce animal use and allow faster testing for hazard assessment. The cells used in this method are maintained and differentiated in media containing foetal calf serum. This animal product is of considerable variation in quality, and individual batches require extensive testing for their applicability in the EST. Moreover, its production involves a large number of foetuses and possible animal suffering. We demonstrate the serum-free medium and feeder cell-free maintenance of the mouse embryonic stem cell line D3 and investigate the use of specific growth factors for induction of cardiac differentiation. Using a combination of bone morphogenetic protein-2, bone morphogenetic protein-4, activin A and ascorbic acid, embryoid bodies efficiently differentiated into contracting myocardium. Additionally, examining levels of intracellular marker proteins by flow cytometry not only confirmed differentiation into cardiomyocytes, but demonstrated significant differentiation into neuronal cells in the same time frame. Thus, this approach might allow for simultaneous detection of developmental effects on both early mesodermal and neuroectodermal differentiation. The serum-free conditions for maintenance and differentiation of D3 cells described here enhance the transferability and standardisation and hence the performance of the EST. Copyright © 2011 Elsevier Ltd. All rights reserved.

  17. Dual color chromogenic in situ hybridization for determination of HER2 status in breast cancer: a large comparative study to current state of the art fluorescence in situ hybridization

    Directory of Open Access Journals (Sweden)

    Mollerup Jens

    2012-02-01

    Full Text Available Abstract Background Chromogenic in situ hybridization (CISH is fast becoming a well established technique for easy and sensitive determination of HER2 gene status in breast cancer. However, for the chromogenic method to achieve status as a safe and reliable technique, the method needs to be validated against already known and validated FISH techniques. Methods Here it is reported from a comparative study where HER2 gene status obtained by HER2 CISH pharmDx™ Kit was compared to HER2 gene status obtained by the FDA approved HER2 FISH pharmDx™ Kit and the PathVysion HER-2 DNA probe Kit. The study included 365 formalin fixed and paraffin-embedded invasive breast cancer tissue specimens collected consecutively at a US reference laboratory. Results The data obtained revealed an overall HER2 status concordance of approximately 98% for comparisons of HER2 CISH pharmDx™ Kit to both HER2 FISH pharmDx™ Kit and PathVysion HER-2 DNA Probe Kit. Conclusions The concordance between results obtained using the recently FDA approved HER2 CISH pharmDx™ Kit with previously FDA approved FISH techniques for HER2 gene status determination indicate that the HER2 CISH pharmDx™ Kit is a reliable chromogenic alternative to fluorescence-based methods.

  18. Dual color chromogenic in situ hybridization for determination of HER2 status in breast cancer: a large comparative study to current state of the art fluorescence in situ hybridization

    Science.gov (United States)

    2012-01-01

    Background Chromogenic in situ hybridization (CISH) is fast becoming a well established technique for easy and sensitive determination of HER2 gene status in breast cancer. However, for the chromogenic method to achieve status as a safe and reliable technique, the method needs to be validated against already known and validated FISH techniques. Methods Here it is reported from a comparative study where HER2 gene status obtained by HER2 CISH pharmDx™ Kit was compared to HER2 gene status obtained by the FDA approved HER2 FISH pharmDx™ Kit and the PathVysion HER-2 DNA probe Kit. The study included 365 formalin fixed and paraffin-embedded invasive breast cancer tissue specimens collected consecutively at a US reference laboratory. Results The data obtained revealed an overall HER2 status concordance of approximately 98% for comparisons of HER2 CISH pharmDx™ Kit to both HER2 FISH pharmDx™ Kit and PathVysion HER-2 DNA Probe Kit. Conclusions The concordance between results obtained using the recently FDA approved HER2 CISH pharmDx™ Kit with previously FDA approved FISH techniques for HER2 gene status determination indicate that the HER2 CISH pharmDx™ Kit is a reliable chromogenic alternative to fluorescence-based methods. PMID:22333181

  19. Sporothrix schenckii Sensu Lato identification in fragments of skin lesion cultured in NNN medium for differential diagnosis of cutaneous leishmaniasis.

    Science.gov (United States)

    Antonio, Liliane de Fátima; Pimentel, Maria Inês Fernandes; Lyra, Marcelo Rosandiski; Madeira, Maria de Fátima; Miranda, Luciana de Freitas Campos; Paes, Rodrigo Almeida; Brito-Santos, Fábio; Carvalho, Maria Helena Galdino Figueredo; Schubach, Armando de Oliveira

    2017-02-01

    Eighty-nine patients with clinical suspicion of leishmaniasis were referred for differential diagnosis. Sporothrix schenckii sensu lato was isolated in Novy-MacNeal-Nicolle + Schneider media in 98% of 64 patients with final diagnosis of sporotrichosis. This medium may be suitable for diagnosis of sporotrichosis in areas where cutaneous leishmaniasis is also endemic. Copyright © 2016 Elsevier Inc. All rights reserved.

  20. Distinct roles of presynaptic dopamine receptors in the differential modulation of the intrinsic synapses of medium-spiny neurons in the nucleus accumbens

    Directory of Open Access Journals (Sweden)

    Schmauss Claudia

    2007-01-01

    Full Text Available Abstract Background In both schizophrenia and addiction, pathological changes in dopamine release appear to induce alterations in the circuitry of the nucleus accumbens that affect coordinated thought and motivation. Dopamine acts principally on medium-spiny GABA neurons, which comprise 95% of accumbens neurons and give rise to the majority of inhibitory synapses in the nucleus. To examine dopamine action at single medium-spiny neuron synapses, we imaged Ca2+ levels in their presynaptic varicosities in the acute brain slice using two-photon microscopy. Results Presynaptic Ca2+ rises were differentially modulated by dopamine. The D1/D5 selective agonist SKF81297 was exclusively facilitatory. The D2/D3 selective agonist quinpirole was predominantly inhibitory, but in some instances it was facilitatory. Studies using D2 and D3 receptor knockout mice revealed that quinpirole inhibition was either D2 or D3 receptor-mediated, while facilitation was mainly D3 receptor-mediated. Subsets of varicosities responded to both D1 and D2 agonists, showing that there was significant co-expression of these receptor families in single medium-spiny neurons. Neighboring presynaptic varicosities showed strikingly heterogeneous responses to DA agonists, suggesting that DA receptors may be differentially trafficked to individual varicosities on the same medium-spiny neuron axon. Conclusion Dopamine receptors are present on the presynaptic varicosities of medium-spiny neurons, where they potently control GABAergic synaptic transmission. While there is significant coexpression of D1 and D2 family dopamine receptors in individual neurons, at the subcellular level, these receptors appear to be heterogeneously distributed, potentially explaining the considerable controversy regarding dopamine action in the striatum, and in particular the degree of dopamine receptor segregation on these neurons. Assuming that post-receptor signaling is restricted to the microdomains of

  1. In vitro cementoblast-like differentiation of postmigratory neural crest-derived p75{sup +} stem cells with dental follicle cell conditioned medium

    Energy Technology Data Exchange (ETDEWEB)

    Wen, Xiujie; Liu, Luchuan; Deng, Manjing; Liu, Rui; Zhang, Li; Nie, Xin, E-mail: dr.xinnie@gmail.com

    2015-09-10

    Cranial neural crest-derived cells (CNCCs) play important role in epithelial–mesenchymal interactions during tooth morphogenesis. However, the heterogeneity of CNCCs and their tendency to spontaneously differentiate along smooth muscle or osteoblast lineages in vitro limit further understanding of their biological properties. We studied the differentiation properties of isolated rat embryonic postmigratory CNCCs, expressing p75 neurotrophin receptor (p75NTR). These p75NTR positive (p75{sup +}) CNCCs, isolated using fluorescence activated cell sorter, exhibited fibroblast-like morphology and characteristics of mesenchymal stem cells. Incubation of p75{sup +} CNCCs in dental follicle cell conditioned medium (DFCCM) combined with dentin non-collagenous proteins (dNCPs), altered their morphological features to cementoblast-like appearance. These cells also showed low proliferative activity, high ALP activity and significantly increased calcified nodule formation. Markers related to mineralization or specific to cementoblast lineage were highly expressed in dNCPs/DFCCM-treated p75{sup +} cells, suggesting their differentiation along cementoblast-like lineage. p75{sup +} stem cells selected from postmigratory CNCCs represent a pure stem cell population and could be used as a stem cell model for in vitro studies due to their intrinsic ability to differentiate to neuronal cells and transform from neuroectoderm to ectomesenchyme. They can provide a potential stem cell resource for tooth engineering studies and help to further investigate mechanisms of epithelial–mesenchymal interactions in tooth morphogenesis. - Highlights: • Cranial neural crest-derived cells (CNCCs) take part in tooth morphogenesis. • positive (p75{sup +}) CNCCs are fibroblast-like and resemble mesenchymal stem cells. • p75{sup +} CNCCs in dental follicle cell medium (DFCCM/dNCP) appear like cementoblasts. • DFCCM/dNCP-treated p75{sup +} cells express cementoblast specific mineralization

  2. Changes in the chromogen properties of the betalaine induced by gamma radiation

    International Nuclear Information System (INIS)

    Rodriguez N, S.; Pinedo S, A.; Amador V, P.; Chacon R, A.; Arcos P, A.; Vega C, H.R.

    2005-01-01

    The changes of coloration of four natural extracts in function of the absorbed dose produced by a gamma rays source of 137 Cs were determined. The natural extracts used contain betalaines that are natural pigments of some plants as the beet about that their name. They are also in abundant form in the fruits (tunas) of some species of the Opuntia genus. The extracts were obtained by maceration, starting from beet and three tuna varieties, and they were stabilized to pH 5.5 The change of coloration it was determined in a visible ultraviolet spectrophotometer by means of the absorbance from the samples to photons of 475, 535 and 600 nm of wavelength. The absorbance to different intervals of time was measured. The relationship between the absorbed dose and the chromogene properties of the pigment, with the intention of using it as possible dosemeter was settled down. (Author)

  3. Combination of retinal pigment epithelium cell-conditioned medium and photoreceptor outer segments stimulate mesenchymal stem cell differentiation toward a functional retinal pigment epithelium cell phenotype.

    Science.gov (United States)

    Huang, Chen; Zhang, Jing; Ao, Mingxin; Li, Ying; Zhang, Chun; Xu, Yonggen; Li, Xuemin; Wang, Wei

    2012-02-01

    Recent studies have suggested that bone marrow-derived mesenchymal stem cells (BMMSCs) are capable of retinal tissue-specific differentiation but not retinal pigment epithelium (RPE) cell-specific differentiation. Photoreceptor outer segments (POS) contribute to RPE development and maturation. However, there has been no standard culture system that fosters the differentiation of BMMSCs into mature RPE cells in vitro. In this study, we investigated if the soluble factors from RPE cells and POS could differentiate BMMSCs into cells having a phenotype characteristic of RPE cells. Rat BMMSCs were separately co-cultured with RPE cells, or they were exposed to either control medium, RPE cell-conditioned medium (RPECM), POS, or a combination of RPECM and POS (RPECM-POS). After 7 days, the cells were analyzed for morphology and the expression of RPE markers (cytokeratin 8, CRALBP, and RPE65) to assess the RPE differentiation. Significantly higher pigment accumulation and increased protein expression of the three markers were seen in cells cultured in RPECM-POS than in other treated cultures. Furthermore, the RPECM-POS-treated cultures displayed ultrastructural features typical of RPE cells, expressed RPE cell functional proteins, and had the capability to phagocytose POS. Together, theses results suggest the combination of RPECM and POS stimulate BMMSCs differentiation toward a functional RPE phenotype. Our results provide the foundation for a new route to RPE regenerative therapy involving BMMSCs. Future work isolating the active agent in RPECM and POS would be useful in therapies for RPE diseases or in developing appropriately pre-differentiated BMMSCs for tissue-engineered RPE reconstruction. Copyright © 2011 Wiley Periodicals, Inc.

  4. Chromogenic in situ hybridization (CISH): a novel alternative in screening archival breast cancer tissue samples for HER-2/neu status

    OpenAIRE

    Madrid, Manuelito A; Lo, Raymundo W

    2004-01-01

    Background Chromogenic in situ hybridization (CISH) is emerging as a practical, cost-effective, and valid alternative to fluorescent in situ hybridization in testing for gene alteration, especially in centers primarily working with immunohistochemistry (IHC). Methods We assessed Her-2/neu alteration using CISH on formalin-fixed paraffin-embedded primary invasive ductal carcinoma tumors in which IHC (CB11 antibody) had previously been performed, and we compared the results with IHC. The 160 se...

  5. New medium used in the differentiation of human pluripotent stem cells to retinal cells is comparable to fetal human eye tissue.

    Science.gov (United States)

    Wang, Xiaobing; Xiong, Kai; Lin, Cong; Lv, Lei; Chen, Jing; Xu, Chongchong; Wang, Songtao; Gu, Dandan; Zheng, Hua; Yu, Hurong; Li, Yan; Xiao, Honglei; Zhou, Guomin

    2015-06-01

    Human pluripotent stem cells (hPSCs) have the potential to differentiate along the retinal lineage. However, most induction systems are dependent on multiple small molecular compounds such as Dkk-1, Lefty-A, and retinoic acid. In the present study, we efficiently differentiated hPSCs into retinal cells using a retinal differentiation medium (RDM) without the use of small molecular compounds. This novel differentiation system recapitulates retinal morphogenesis in humans, i.e. hPSCs gradually differentiate into optic vesicle-shaped spheres, followed by optic cup-shaped spheres and, lastly, retinal progenitor cells. Furthermore, at different stages, hPSC-derived retinal cells mirror the transcription factor expression profiles seen in their counterparts during human embryogenesis. Most importantly, hinge epithelium was found between the hPSC-derived neural retina (NR) and retinal pigment epithelium (RPE). These data suggest that our culture system provides a new method for generating hPSC-derived retinal cells that, for the first time, might be used in human transplantation. Copyright © 2015 Elsevier Ltd. All rights reserved.

  6. Mycobacterium eburneum sp. nov., a non-chromogenic, fast-growing strain isolated from sputum.

    Science.gov (United States)

    Nouioui, Imen; Carro, Lorena; Teramoto, Kanae; Igual, José M; Jando, Marlen; Del Carmen Montero-Calasanz, Maria; Sutcliffe, Iain; Sangal, Vartul; Goodfellow, Michael; Klenk, Hans-Peter

    2017-09-01

    A polyphasic study was undertaken to establish the taxonomic position of a non-chromogenic, rapidly growing Mycobacterium strain that had been isolated from sputum. The strain, CECT 8775T, has chemotaxonomic and cultural properties consistent with its classification in the genus Mycobacterium and was distinguished from the type strains of closely related mycobacterial species, notably from Mycobacterium paraense DSM 46749T, its nearest phylogenetic neighbour, based on 16S rRNA, hsp65 and rpoB gene sequence data. These organisms were also distinguished by a broad range of chemotaxonomic and phenotypic features and by a digital DNA-DNA relatedness value of 22.8 %. Consequently, the strain is considered to represent a novel species of Mycobacterium for which the name Mycobacterium eburneum sp. nov is proposed; the type strain is X82T (CECT 8775T=DSM 44358T).

  7. 21 CFR 866.2320 - Differential culture medium.

    Science.gov (United States)

    2010-04-01

    ...) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Microbiology Devices § 866.2320 Differential culture... occurred. Test results aid in the diagnosis of disease and also provide epidemiological information on...

  8. Application of 2-Trichloromethylbenzimidazole in Analytical Chemistry: A Highly Selective Chromogenic Reagent for Thin-Layer Chromatography and Some Other Analytical Uses

    Directory of Open Access Journals (Sweden)

    Leszek Konopski

    2012-01-01

    Full Text Available 2-Trichloromethylbenzimidazole (TCMB was used as a chromogenic reagent in organic or inorganic analysis, mainly in thin-layer chromatography (TLC. In reactions of TCMB with some heteroaromatic nitrogen containing compounds, such as azines, azoles and benzazoles, a formation of high colored products occurred. For azines, the chromogenic reaction was highly regioselective, since the both adjacent α-positions versus the nitrogen atom(s must not be substituted. A TLC method of detection was developed. Thirty azines, azoles, and benzazoles were detected at the detection limit 10 ng to 1 μg. This method was also applied for detection of heteroaromatic pesticides, and the attempts to construct active and passive dosimeters for nicotine were made. In a prechromatographic reaction of aromatic o-diamines with methyl trichloroacetimidate, TCMB or its derivatives were formed in situ. Followed by TLC and visualization in pyridine vapors, this procedure was applied for detection of o-phenylenediamine derivatives. The reaction product of TCMB and pyridine (LI Complex was identified and fully characterized. Two different reaction mechanisms: with electron deficient basic heteroaromatic compounds, like pyridine, and with more acidic compounds, for example, pyrrole, were discussed. In aqueous solutions, the LI Complex may be also used as a new indicator for complexometric, adsorption and acid-base titration of inorganic compounds.

  9. A rapid kinetic chromogenic method for quantification of bacterial endotoxins in lyophilized reagents for labeling with 99mTc radiopharmaceuticals

    International Nuclear Information System (INIS)

    Fukumori, Neuza T.O.; Campos, Domingos G.; Silva, Laercio; Fernandes, Adriana V.; Mengatti, Jair; Silva, Constancia P.G.; Matsuda, Margareth M.N.

    2009-01-01

    A rapid quantitative kinetic chromogenic test in an automated Portable Test System (PTS) has been developed for determination of bacterial endotoxins in water, in-process and end-products using the Limulus amebocyte lysate (LAL). The aim of this work was to validate the method for lyophilized reagents for labeling with 99m Tc radiopharmaceuticals with no interfering factors. Experiments were performed in three consecutive batches of the lyophilized reagents Methylenediphosphonic Acid (MDP) and Pyrophosphate (PYRO) produced at IPEN-CNEN/ SP using the PTS from Endosafe, Inc. TM , Charleston, SC. The Maximum Valid Dilution (MVD) was calculated to establish the extent of dilution to avoid interfering test conditions (MVD=500). Better results were obtained above 1:20 dilution factor for MDP and 1:100 for PYRO. The parameters of coefficient correlation (R) -0.980, RPPC between 50 - 200% and coefficient variation (CV) of the samples less than 25% were satisfied and the endotoxin concentration was lower than the lowest concentration of the standard curve (0.05 EU mL -1 ), therefore less than the established limit in pharmacopoeias. The PTS is a rapid, simple and accurate technique using the quantitative kinetic chromogenic method for bacterial endotoxin determination. For this reason, it is very practical in the radiopharmaceutical area and it trends to be the method of choice for the pyrogen test. For MDP and PYRO, the validation was successfully performed. (author)

  10. Research on the Phenotypic Characterization of Mrsa Strains Isolated from Animals

    Directory of Open Access Journals (Sweden)

    Iulia Maria BUCUR

    2017-05-01

    Full Text Available Keywords: chromogen, methicillin, MRSA, resistance Introduction: Currently, both in staphylococci isolated from animals with different diseases, as well as in humans, the MRSA strains (Methicillin Resistant S. aureus are monitored, as the methicillin resistance is associated with the resistance to other antibiotic groups. Methicillin resistance is encoded by mec staphylococcal chromosomal cassettes (SCCmec, which are islands of resistance. These strains can be identified by molecular biology tests and tests that reveal several phenotypic characteristics. The research was made in order to characterize and identify phenotypically the MRSA staphylococci strains isolated from animals. Materials and Methods: Researches were made on 240 coagulase positive and coagulase negative strains of staphylococci. Mannitol fermentation was tested on Champan medium, free coagulase was revealed on Baird-Parker medium and to identify S. aureus subsp. aureus was used the chromogenic medium Chromatic Staph. Methicillin-resistant strains were detected by disc diffusion method, using biodiscs with methicillin, oxacillin and cefoxitin. Also, to identify the MRSA strains, was used the chromogenic medium Chromatic MRSA. Results: The isolates were positive to mannitol and produced complete haemolysis or were unhaemolytic. A total of 44 strains produced free coagulase on Baird-Parker medium, considered coagulase positive strains, while 196 were coagulase negative strains. The isolates conducted differently to methicillin: 22,08% of strains were resistant, 51,25% of strains were susceptible and 26,66% had intermediate resistance, while the resistant strains to oxacillin were 42,91%. The increased frequency of methicillin-resistant strains of staphylococci and, particularly, MRSA strains, determined using the cefoxitin disk diffusion test, which is more reliable than methicillin and oxacillin. On the MRSA chromogenic medium, the methicillin-resistant strains of staphylococci

  11. H2O2-responsive liposomal nanoprobe for photoacoustic inflammation imaging and tumor theranostics via in vivo chromogenic assay.

    Science.gov (United States)

    Chen, Qian; Liang, Chao; Sun, Xiaoqi; Chen, Jiawen; Yang, Zhijuan; Zhao, He; Feng, Liangzhu; Liu, Zhuang

    2017-05-23

    Abnormal H 2 O 2 levels are closely related to many diseases, including inflammation and cancers. Herein, we simultaneously load HRP and its substrate, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), into liposomal nanoparticles, obtaining a Lipo@HRP&ABTS optical nanoprobe for in vivo H 2 O 2 -responsive chromogenic assay with great specificity and sensitivity. In the presence of H 2 O 2 , colorless ABTS would be converted by HRP into the oxidized form with strong near-infrared (NIR) absorbance, enabling photoacoustic detection of H 2 O 2 down to submicromolar concentrations. Using Lipo@HRP&ABTS as an H 2 O 2 -responsive nanoprobe, we could accurately detect the inflammation processes induced by LPS or bacterial infection in which H 2 O 2 is generated. Meanwhile, upon systemic administration of this nanoprobe we realize in vivo photoacoustic imaging of small s.c. tumors (∼2 mm in size) as well as orthotopic brain gliomas, by detecting H 2 O 2 produced by tumor cells. Interestingly, local injection of Lipo@HRP&ABTS further enables differentiation of metastatic lymph nodes from those nonmetastatic ones, based on their difference in H 2 O 2 contents. Moreover, using the H 2 O 2 -dependent strong NIR absorbance of Lipo@HRP&ABTS, tumor-specific photothermal therapy is also achieved. This work thus develops a sensitive H 2 O 2 -responsive optical nanoprobe useful not only for in vivo detection of inflammation but also for tumor-specific theranostic applications.

  12. Comparison of Chromogenic In Situ Hybridization and Fluorescence In Situ Hybridization for the Evaluation of MDM2 Amplification in Adipocytic Tumors.

    Science.gov (United States)

    Mardekian, Stacey K; Solomides, Charalambos C; Gong, Jerald Z; Peiper, Stephen C; Wang, Zi-Xuan; Bajaj, Renu

    2015-11-01

    Atypical lipomatous tumor/well-differentiated liposarcoma (ALT-WDLPS) and dedifferentiated liposarcoma (DDLPS) are characterized cytogenetically by a 12q13-15 amplification involving the mouse double minute 2 (MDM2) oncogene. Fluorescence in situ hybridization (FISH) is used frequently to detect this amplification and aid with the diagnosis of these entities, which is difficult by morphology alone. Recently, bright-field in situ hybridization techniques such as chromogenic in situ hybridization (CISH) have been introduced for the determination of MDM2 amplification status. The present study compared the results of FISH and CISH for detecting MDM2 amplification in 41 cases of adipocytic tumors. Amplification was defined in both techniques as a MDM2/CEN12 ratio of 2 or greater. Eleven cases showed amplification with both FISH and CISH, and 26 cases showed no amplification with both methods. Two cases had discordant results between CISH and FISH, and two cases were not interpretable by CISH. CISH is advantageous for allowing pathologists to evaluate the histologic and molecular alterations occurring simultaneously in a specimen. Moreover, CISH is found to be more cost- and time-efficient when used with automation, and the signals do not quench over time. CISH technique is a reliable alternative to FISH in the evaluation of adipocytic tumors for MDM2 amplification. © 2014 Wiley Periodicals, Inc.

  13. ESBL Detection: Comparison of a Commercially Available Chromogenic Test for Third Generation Cephalosporine Resistance and Automated Susceptibility Testing in Enterobactericeae.

    Directory of Open Access Journals (Sweden)

    Mohamed Ramadan El-Jade

    Full Text Available Rapid detection and reporting of third generation cephalosporine resistance (3GC-R and of extended spectrum betalactamases in Enterobacteriaceae (ESBL-E is a diagnostic and therapeutic priority to avoid inefficacy of the initial antibiotic regimen. In this study we evaluated a commercially available chromogenic screen for 3GC-R as a predictive and/or confirmatory test for ESBL and AmpC activity in clinical and veterinary Enterobacteriaceae isolates. The test was highly reliable in the prediction of cefotaxime and cefpodoxime resistance, but there was no correlation with ceftazidime and piperacillin/tazobactam minimal inhibitory concentrations. All human and porcine ESBL-E tested were detected with exception of one genetically positive but phenotypically negative isolate. By contrast, AmpC detection rates lay below 30%. Notably, exclusion of piperacillin/tazobactam resistant, 3GC susceptible K1+ Klebsiella isolates increased the sensitivity and specificity of the test for ESBL detection. Our data further imply that in regions with low prevalence of AmpC and K1 positive E. coli strains chromogenic testing for 3GC-R can substitute for more time consuming ESBL confirmative testing in E. coli isolates tested positive by Phoenix or VITEK2 ESBL screen. We, therefore, suggest a diagnostic algorithm that distinguishes 3GC-R screening from primary culture and species-dependent confirmatory ESBL testing by βLACTATM and discuss the implications of MIC distribution results on the choice of antibiotic regimen.

  14. Direct identification and recognition of yeast species from clinical material by using albicans ID and CHROMagar Candida plates.

    OpenAIRE

    Baumgartner, C; Freydiere, A M; Gille, Y

    1996-01-01

    Two chromogenic media, Albicans ID and CHROMagar Candida agar plates, were compared with a reference medium, Sabouraud-chloramphenicol agar, and standard methods for the identification of yeast species. This study involved 951 clinical specimens. The detection rates for the two chromogenic media for polymicrobial specimens were 20% higher than that for the Sabouraud-chloramphenicol agar plates. The rates of identification of Candida albicans for Albicans ID and CHROMagar Candida agar plates w...

  15. Prospective Comparison of a New Chromogenic Medium, MRSASelect, to CHROMagar MRSA and Mannitol-Salt Medium Supplemented with Oxacillin or Cefoxitin for Detection of Methicillin-Resistant Staphylococcus aureus

    Science.gov (United States)

    Stoakes, Luba; Reyes, Romina; Daniel, Janis; Lennox, Gwen; John, Michael A.; Lannigan, Robert; Hussain, Zafar

    2006-01-01

    MRSASelect agar was compared to CHROMagar, mannitol-salt agar with oxacillin, and mannitol-salt agar with cefoxitin (MSA-CFOX) for the isolation of methicillin-resistant Staphylococcus aureus (MRSA). The sensitivities and specificities were 97.3% and 99.8%, 82.9% and 99.1%, 80.2% and 79%, and 99.1% and 84.8%, respectively. MSA-CFOX and MRSASelect had a high sensitivity. MRSASelect, however, was more specific and proved to be a more reliable and rapid medium for the detection of MRSA. PMID:16455933

  16. Quasifree η photoproduction from nuclei and medium modifications of resonances

    International Nuclear Information System (INIS)

    Ventel, B.I.S. van der; Abu-Raddad, L.J.; Hillhouse, G.C.

    2003-01-01

    This paper establishes the case that the process of quasifree η photoproduction from nuclei is an important tool to study medium modifications and changes to the elementary process γN→ηN in the nuclear medium. We investigate the sensitivity of the differential cross section, recoil nucleon polarization, and the photon asymmetry to changes in the elementary amplitude, medium modifications of the resonance (S 11 ,D 13 ) masses, as well as nuclear target effects. All calculations are performed within a relativistic plane-wave impulse approximation formalism resulting in analytical expressions for all observables. Our results indicate that polarization observables are largely insensitive to nuclear target effects. Depending on the type of coupling, the spin observables do display a sensitivity to the magnitude of the ηNN coupling constant. The polarization observables are identified to be the prime candidates to investigate the background processes and their medium modifications in the elementary process such as the D 13 resonance. Moreover, as a consequence of the large dominance in the differential cross section of the S 11 resonance, the quasifree differential cross section provides an exceptional instrument to study medium modifications to the S 11 resonance in such a manner that helps to distinguish between various models that attempt to understand the S 11 resonance and its distinctive position as the lowest lying negative parity state in the baryon spectrum

  17. The Study of Birefringent Homogenous Medium with Geometric Phase

    International Nuclear Information System (INIS)

    Banerjee, Dipti

    2010-12-01

    The property of linear and circular birefringence at each point of the optical medium has been evaluated here from differential matrix N using the Jones calculus. This matrix lies on the OAM sphere for l = 1 orbital angular momentum. The geometric phase is developed by twisting the medium uniformly about the direction of propagation of the light ray. The circular birefringence of the medium, is visualized through the solid angle and the angular twist per unit thickness of the medium, k, that is equivalent to the topological charge of the optical element. (author)

  18. Fast methods of fungal and bacterial identification. MALDI-TOF mass spectrometry, chromogenic media.

    Science.gov (United States)

    Siller-Ruiz, María; Hernández-Egido, Sara; Sánchez-Juanes, Fernando; González-Buitrago, José Manuel; Muñoz-Bellido, Juan Luis

    2017-05-01

    MALDI-TOF mass spectrometry is now a routine resource in Clinical Microbiology, because of its speed and reliability in the identification of microorganisms. Its performance in the identification of bacteria and yeasts is perfectly contrasted. The identification of mycobacteria and moulds is more complex, due to the heterogeneity of spectra within each species. The methodology is somewhat more complex, and expanding the size of species libraries, and the number of spectra of each species, will be crucial to achieve greater efficiency. Direct identification from blood cultures has been implemented, since its contribution to the management of severe patients is evident, but its application to other samples is more complex. Chromogenic media have also contributed to the rapid diagnosis in both bacteria and yeast, since they accelerate the diagnosis, facilitate the detection of mixed cultures and allow rapid diagnosis of resistant species. Copyright © 2017 Elsevier España, S.L.U. and Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica. All rights reserved.

  19. Derivation, characterization and retinal differentiation of induced ...

    Indian Academy of Sciences (India)

    2013-01-11

    Jan 11, 2013 ... ing the differentiating medium with a RPE-cell-line-condi- tioned medium. 2. ... medium and plated into four 100 mm dishes and the cultures ... mitotically inactivated by applying 5000 rad of radiation in a .... Temperature °C.

  20. Effect of partial heating at mid of vertical plate adjacent to porous medium

    Science.gov (United States)

    Mulla, Mohammed Fahimuddin; Pallan, Khalid. M.; Al-Rashed, A. A. A. A.

    2018-05-01

    Heat and mass transfer in porous medium due to heating of vertical plate at mid-section is analyzed for various physical parameters. The heat and mass transfer in porous medium is modeled with the help of momentum, energy and concentration equations in terms of non-dimensional partial differential equations. The partial differential equations are converted into simpler form of algebraic equations with the help of finite element method. A computer code is developed to assemble the matrix form of algebraic equations into global matrices and then to solve them in an iterative manner to obtain the temperature, concentration and streamline distribution inside the porous medium. It is found that the heat transfer behavior of porous medium heated at middle section is considerably different from other cases.

  1. Y-chromosome status identification suggests a recipient origin of posttransplant non-small cell lung carcinomas: chromogenic in situ hybridization analysis.

    Science.gov (United States)

    Chen, Wei; Brodsky, Sergey V; Zhao, Weiqiang; Otterson, Gregory A; Villalona-Calero, Miguel; Satoskar, Anjali A; Hasan, Ayesha; Pelletier, Ronald; Ivanov, Iouri; Ross, Patrick; Nadasdy, Tibor; Shilo, Konstantin

    2014-05-01

    Owing to the need of lifelong immunosuppression, solid-organ transplant recipients are known to have an increased risk of posttransplant malignancies including lung cancer. Posttransplant neoplastic transformation of donor-derived cells giving rise to hematopoietic malignancies, Kaposi sarcoma, and basal cell carcinoma in nongraft tissues has been reported. The goal of this study was to assess the cell origin (donor versus recipient derived) of posttransplant non-small cell lung carcinomas (NSCLCs) in kidney and heart transplant recipients. An institutional database search identified 2557 kidney and heart transplant recipients in 8 consecutive years. Among this cohort, 20 (0.8%) renal and 18 (0.7%) heart transplant recipients developed NSCLC. The study cohort comprised 6 of 38 NSCLCs arising in donor-recipient sex-mismatched transplant patients. The tumor cell origin was evaluated by chromogenic in situ hybridization with Y-chromosome probe on formalin-fixed, paraffin-embedded tissues. Y-chromosome was identified in 97% ± 1% (range from 92% to 99%) of all types of nucleated cells in male control tissues. In all 5 NSCLCs from male recipients of female donor organ, Y-chromosome was identified in 97% ± 2% (range from 92% to 100%) of tumor cells, statistically equivalent to normal control (P recipient of male kidney. These findings suggest a recipient derivation of NSCLC arising in kidney and heart transplant recipients. A combination of histologic evaluation and chromogenic in situ hybridization with Y-chromosome analysis allows reliable determination of tissue origin in sex-mismatched solid-organ transplant recipients and may aid in management of posttransplant malignancy in such cases. Copyright © 2014 Elsevier Inc. All rights reserved.

  2. Mixed Convection Flow along a Stretching Cylinder in a Thermally Stratified Medium

    Directory of Open Access Journals (Sweden)

    Swati Mukhopadhyay

    2012-01-01

    Full Text Available An analysis for the axisymmetric laminar boundary layer mixed convection flow of a viscous and incompressible fluid towards a stretching cylinder immersed in a thermally stratified medium is presented in this paper. Similarity transformation is employed to convert the governing partial differential equations into highly nonlinear ordinary differential equations. Numerical solutions of these equations are obtained by a shooting method. It is found that the heat transfer rate at the surface is lower for flow in a thermally stratified medium compared to that of an unstratified medium. Moreover, both the skin friction coefficient and the heat transfer rate at the surface are larger for a cylinder compared to that for a flat plate.

  3. JC virus chromogenic in situ hybridization in brain biopsies from patients with and without PML.

    Science.gov (United States)

    Procop, Gary W; Beck, Rose C; Pettay, James D; Kohn, Debra J; Tuohy, Marion J; Yen-Lieberman, Belinda; Prayson, Richard A; Tubbs, Raymond R

    2006-06-01

    Progressive multifocal leukoencephalopathy (PML) is a demyelinating disease of the central nervous system caused by the JC polyoma virus. Electron microscopy and immunohistochemistry are the traditional methods of confirming the presence of the virus in brain biopsies from these patients. We studied the brain biopsies from 7 patients with PML and 6 patients without PML with chromogenic in situ hybridization (CISH) for the JC polyoma virus using a commercially available probe. The biopsies from the patients with the PML cases were proven to contain the JC polyoma virus by traditional and molecular methods. The CISH findings were compared with the known state of infection. All (7/7) of the biopsies from patients with PML were positive for the presence of polyoma virus by CISH, whereas the biopsies from patients without PML were uniformly negative. CISH seems to be a useful tool for the detection of the JC virus in brain biopsies from patients with PML, and is more accessible because a commercial probe is available.

  4. Determination of free cisplatin in medium by differential pulse polarography after ultrasound and cisplatin treatment of a cancer cell culture

    International Nuclear Information System (INIS)

    Bernard, Vladan; Skorpikova, Jirina; Mornstein, Vojtech; Fojt, Lukas

    2011-01-01

    The in vitro study was carried out for detection of the cisplatin in free form and in culture medium, depending on various conditions of sonodynamic human ovarian cancer cells A2780 treatment by differential pulse polarography (DPP). For sonodynamic treatment, we used cisplatin alone and combined cisplatin/ultrasound treatments. The ultrasound exposure intensity of 1.0 and 2.0 Wcm 2 in far field for incubation periods 1, 24 and 48 h was used. The parameters of DPP measurements were - 1 s drop time, 5 mV.s -1 voltage scan rate, 50 mV modulation amplitude and negative scanning direction; platinum wire served as counter electrode and Ag|AgCl|3 M KCI as reference electrode. The results showed the dependence of free platinum quantities in culture medium on incubation time and treatment protocol. We found difference in concentration of free cisplatin between conventional application of cisplatin and sonodynamic treatment. The sonodynamic combined treatment of cisplatin and ultrasound field showed a higher cisplatin content in the culture medium than cisplatin treatment alone; a difference of 20% was observed for incubation time 48 h. The results also showed the influence of a time sequence of ultrasound and cytostatics in the sonodynamic treatment. The highest amount of free cisplatin in the solution was found for primary application of cisplatin and the subsequent ultrasound exposure. The quantity of free cisplatin increased with time, namely for time intervals 1-24 h. There was no difference between the DPP signal of cisplatin in reaction mixture containing cells in small quantities and micro-filtered mixture without cells. Thus, the DPP method is suitable for the detection and quantification of free cisplatin in the culture medium of cell suspension. Ultrasound field can be important factor during cytostatic therapy. (author)

  5. DYNAMIC DEFORMATION THE VISCOELASTIC TWOCOMPONENT MEDIUM

    Directory of Open Access Journals (Sweden)

    V. S. Polenov

    2015-01-01

    Full Text Available Summary. In the article are scope harmonious warping of the two-component medium, one component which are represent viscoelastic medium, hereditary properties which are described by the kernel aftereffect Abel integral-differential ratio BoltzmannVolterr, while second – compressible liquid. Do a study one-dimensional case. Use motion equation of two-component medium at movement. Look determination system these equalization in the form of damped wave. Introduce dimensionless coefficient. Combined equations happen to homogeneous system with complex factor relatively waves amplitude in viscoelastic component and in fluid. As a result opening system determinant receive biquadratic equation. Elastic operator express through kernel aftereffect Abel for space Fourier. With the help transformation and symbol series biquadratic equation reduce to quadratic equation. Come to the conclusion that in two-component viscoelastic medium exist two mode sonic waves. As a result solution of quadratic equation be found description advance of waves sonic in viscoelastic two-component medium, which physical-mechanical properties represent complex parameter. Velocity determination advance of sonic waves, attenuation coefficient, mechanical loss tangent, depending on characteristic porous medium and circular frequency formulas receive. Graph dependences of description advance of waves sonic from the temperature logarithm and with the fractional parameter γ are constructed.

  6. Detection of Tritrichomonas foetus and Pentatrichomonas hominis in intestinal tissue specimens of cats by chromogenic in situ hybridization.

    Science.gov (United States)

    Mostegl, Meike M; Wetscher, Andreas; Richter, Barbara; Nedorost, Nora; Dinhopl, Nora; Weissenböck, Herbert

    2012-02-10

    In this retrospective study 102 cats were analyzed for the presence of trichomonads in intestinal tissue sections using chromogenic in situ hybridization (CISH). Two intestinal trichomonad species are described in cats: Pentatrichomonas hominis and Tritrichomonas foetus. While P. hominis is considered a mere commensal, T. foetus has been found to be the causative agent of feline large-bowel diarrhea. For the detection of both agents within intestinal tissue CISH assays using three different probes were performed. In the first CISH run a probe specific for all relevant members of the order Trichomonadida (OT probe) was used. In a second CISH run all positive samples were further examined on three consecutive tissue sections using the OT probe, a probe specific for the family of Tritrichomonadidae (Tritri probe) and a newly designed probe specifically detecting P. hominis (Penta hom probe). In total, four of the 102 cats were found to be positive with the OT probe. Thereof, one cat gave a positive reaction with the P. hominis probe and three cats were positive with the T. foetus probe. All Trichomonas-positive cats were pure-bred and between 8 and 32 weeks of age. In one cat positive for T. foetus large amounts of parasites were found in the gut lumen and invading the intestinal mucosa. The species of the detected trichomonads were confirmed by polymerase chain reaction and nucleotide sequencing of a part of the 18S ribosomal RNA gene. In this study, the usefulness of CISH to detect intestinal trichomonads within feline tissue samples was shown. Additionally, the specific detection of P. hominis using CISH was established. Generally, it was shown that CISH is well suited for detection and differentiation of trichomonosis in retrospective studies using tissue samples. Copyright © 2011 Elsevier B.V. All rights reserved.

  7. Coagulase-negative staphylococci from non-mastitic bovine mammary gland: characterization of Staphylococcus chromogenes and Staphylococcus haemolyticus by antibiotic susceptibility testing and pulsed-field gel electrophoresis.

    Science.gov (United States)

    Pate, Mateja; Zdovc, Irena; Avberšek, Jana; Ocepek, Matjaž; Pengov, Andrej; Podpečan, Ožbalt

    2012-05-01

    During routine microbiological examination of milk samples from dairy cows without clinical signs of mastitis, quarter milk samples of 231 dairy cows from 12 herds were investigated for the presence of coagulase-negative staphylococci (CNS). The isolates were identified on the basis of colony morphology, Gram staining, catalase and coagulase test and the commercial kit, API Staph. CNS was detected in 29% (67/231) of the cows. A total of seven CNS species were identified with the most prevalent being Staphylococcus (Staph.) chromogenes (30%) and Staph. haemolyticus (28·8%), followed by Staph. simulans (11·2%), Staph. xylosus (11·2%), Staph. epidermidis (7·5%), Staph. hyicus (6·3%) and Staph. sciuri (5%). The predominant species, Staph. chromogenes and Staph. haemolyticus, were further characterized by antibiotic susceptibility testing using the agar disc diffusion method (Kirby-Bauer) and by pulsed-field gel electrophoresis (PFGE). Considerable resistance to ampicillin and penicillin was observed in both species. Isolates with identical or highly similar PFGE profiles were detected at the herd level despite a marked heterogeneity seen for both species. On the basis of somatic cell count, absence of clinical signs of inflammation and heterogeneity of genotypes, we assume that CNS isolated in this study could not be considered as important causative agents of the bovine mammary gland inflammation.

  8. Diagnostic properties of three conventional selective plating media for selection of Bacillus cereus, B. thuringiensis and B. weihenstephanensis

    DEFF Research Database (Denmark)

    Hendriksen, Niels Bohse; Hansen, Bjarne Munk

    2011-01-01

    The aim of this study was to assess the diagnostic properties of the two selective plating media and a chromogenic medium for identification of Bacillus cereus. The 324 isolates were B. cereus (37%), Bacillus weihenstephanensis (45%) or Bacillus thuringiensis (18%), as identified by a new...... combination of techniques. All isolates were growing on mannitol–egg yolk–polymyxin agar (MYP), and they did not form acid from mannitol. However, a significant lower number of B. thuringiensis isolates did not show lecithinase activity. All isolates were also growing on polymyxin–egg yolk...... recommended selective plating media MYP and PEMBA for detection of B. cereus group bacteria both have their limitations for identification of some B. cereus, B. weihenstephanensis or B. thuringiensis. However, MYP is preferable compared to PEMBA. The chromogenic medium has its own advantages and limitations...

  9. Correlation and comparison of immunohistochemistry for HER2/neu, using the antibody SP3 and chromogenic in situ hybridization in breast carcinomas samples

    Directory of Open Access Journals (Sweden)

    Franciele F. Wolf

    2015-12-01

    Full Text Available ABSTRACT Introduction: Advances in the field of molecular biology have provided the differentiation of molecular subtypes of breast tumors, providing better prognosis and important tools for the treatment of patients with breast cancer. Among these subtypes, the changes in the human epidermal growth factor receptor 2 gene (HER2/neu, increase its copy number and generating HER2 protein amplification. Studies show that patients with breast cancer HER2/neu amplified tend to relapse earlier and have shorter survival time, the monoclonal antibody Trastuzumab is the therapy indicated. The eligibility of patients for therapy is initially made by the immunohistochemistry (IHC technique, which evaluates the expression level of the HER2 protein. After this evaluation, the cases with equivocal diagnosis (score 2+, are referred to a more accurate technique, the chromogenic in situ hybridization (CISH. Objective: To analyze the sensitivity and specificity of the antibody SP3, and determine their level of agreement with the CISH technique. Material and methods: Retrospective study in the database of the anatomy-pathology laboratory, in CISH tests reports for HER2/neu. Conclusion: The results revealed that clone SP3 showed 100% specificity and 92% sensitivity. IHC reveals variability in its results; however, it is known that the technique is an important tool in the daily routine of laboratories, contributing to the initial screening of patients with breast cancer, which later showed satisfactory results when compared with the CISH technique.

  10. Electromagnetic Radiation in a Uniformly Moving, Homogeneous Medium

    DEFF Research Database (Denmark)

    Johannsen, Günther

    1972-01-01

    A new method of treating radiation problems in a uniformly moving, homogeneous medium is presented. A certain transformation technique in connection with the four-dimensional Green's function method makes it possible to elaborate the Green's functions of the governing differential equations...

  11. Fem Formulation of Heat Transfer in Cylindrical Porous Medium

    Science.gov (United States)

    Azeem; Khaleed, H. M. T.; Soudagar, Manzoor Elahi M.

    2017-08-01

    Heat transfer in porous medium can be derived from the fundamental laws of flow in porous region ass given by Henry Darcy. The fluid flow and energy transport inside the porous medium can be described with the help of momentum and energy equations. The heat transfer in cylindrical porous medium differs from its counterpart in radial and axial coordinates. The present work is focused to discuss the finite element formulation of heat transfer in cylindrical porous medium. The basic partial differential equations are derived using Darcy law which is the converted into a set of algebraic equations with the help of finite element method. The resulting equations are solved by matrix method for two solution variables involved in the coupled equations.

  12. Differentiated-effect shims for medium field levels and saturation

    International Nuclear Information System (INIS)

    Richie, A.

    1976-01-01

    The arrangement of shims on the upstream and downstream ends of magnets may be based on the independent effects of variations in the geometric length and degree of saturation at the edges of the poles. This technique can be used to match the bending strength of an accelerator's magnets at two field levels (medium fields and maximum fields) and thus save special procedures (mixing the laminations, local compensation for errors by arranging the magnets in the appropriate order) and special devices (for instance, correcting dipoles) solely for correcting bending strengths at low field levels. (Auth.)

  13. Monosomy 3 by chromogenic in situ hybridization (CISH) in Iranian patients with uveal melanoma.

    Science.gov (United States)

    Naseripour, Masood; Mehrazma, Mitra; Pourmatin, Rama; Kashkouli, Mohsen Bahmani; Sedaghat, Ahad; Gheytanchi, Elmira

    2015-01-01

    The aim of this study was to investigate the rate of monosomy 3 by CISH technique in Iranian patients with uveal melanoma (UM) and its correlation with clinical and histopathological features. Archival formalin fixed, paraffin-embedded material from 50 patients who had undergone enucleation for large uveal melanomas was obtained. Monosomy of chromosome 3 alteration by chromogenic in situ hybridization (CISH) was investigated. Clinical and histopathological features of tumors were collected. The patients had a mean age of 56.6±7.6 years. Mean basal diameter and thickness of tumors were 14.1 mm and 10.2 mm, respectively. Four patients (8%) were identified to harbor monosomy of chromosome 3. In the mean follow-up of 5.3 years (range, 3.2-9.5 y), only one case with monosomy 3 died of UM metastasis. The most common type of cellularity was mixed cell (86%). There was not any statistically significant correlation between monosomy of chromosome 3 and type of cellularity, ciliary body involvement, and largest basal diameter. The low rate of monosomy chromosome 3 and the consequent low rate of mortality may be indicative of good prognosis in Iranian patients with uveal melanoma.

  14. Application of Photoshop and Scion Image analysis to quantification of signals in histochemistry, immunocytochemistry and hybridocytochemistry.

    Science.gov (United States)

    Tolivia, Jorge; Navarro, Ana; del Valle, Eva; Perez, Cristina; Ordoñez, Cristina; Martínez, Eva

    2006-02-01

    To describe a simple method to achieve the differential selection and subsequent quantification of the strength signal using only one section. Several methods for performing quantitative histochemistry, immunocytochemistry or hybridocytochemistry, without use of specific commercial image analysis systems, rely on pixel-counting algorithms, which do not provide information on the amount of chromogen present in the section. Other techniques use complex algorithms to calculate the cumulative signal strength using two consecutive sections. To separate the chromogen signal we used the "Color range" option of the Adobe Photoshop program, which provides a specific file for a particular chromogen selection that could be applied on similar sections. The measurement of the chromogen signal strength of the specific staining is achieved with the Scion Image software program. The method described in this paper can also be applied to simultaneous detection of different signals on the same section or different parameters (area of particles, number of particles, etc.) when the "Analyze particles" tool of the Scion program is used.

  15. Contribution to the determination of the double angular and energy differential neutron albedo. Application to the propagation in lacunar medium

    International Nuclear Information System (INIS)

    Litaize, O.

    2000-01-01

    The goal of this thesis is to study the neutron propagation by reflection from lacunar medium interfaces. The most efficient method to calculate this type of propagation is to use the concept of albedo. Actual version of NARCISSE code uses a simple formulation of angular differential albedos and so, can only treat single reflections. Multiple reflections treatment needs the knowledge of neutron spectrum after reflection. This energetic information is contained in double angular and energy differential albedos. The first step of this study consists to generate these albedos for various materials. Several methods have been tested and the Monte Carlo method was retained. A new estimator has been developed and validated in the Mote Carlo transport code TRIPOLI-4. It computes, during the simulation of the neutron history, the angular and energy reflection probability at each collision site. The second step consists to generate an interpolation scheme and albedo libraries for various materials. A new version of NARCISSE was developed to use these libraries and the interpolation module. Spectrum and dose rates comparisons were made between codes to validate these albedos. The neutron propagation by multiple reflections can be studied now, by using this new version of Narcisse. (author)

  16. Role of Relationship Marketing in Small and Medium-Sized Entreprises

    OpenAIRE

    Ružica Butigan; Ivana Mahnić

    2011-01-01

    Along with marketing, small and medium enterprises (SMEs) are commonly associated in literature with relationship marketing which results in marketing networks. This paper examines the specific characteristics that differentiate large companies from small and medium-sized enterprises, and the reasons that prevent SMEs from engaging in traditional marketing within the scope of marketing mix. The paper also shows that the key characteristic which distinguishes small from large companies is a pr...

  17. The Role of Paracrine and Autocrine Signaling in the Early Phase of Adipogenic Differentiation of Adipose-derived Stem Cells

    Science.gov (United States)

    Hemmingsen, Mette; Vedel, Søren; Skafte-Pedersen, Peder; Sabourin, David; Collas, Philippe; Bruus, Henrik; Dufva, Martin

    2013-01-01

    Introduction High cell density is known to enhance adipogenic differentiation of mesenchymal stem cells, suggesting secretion of signaling factors or cell-contact-mediated signaling. By employing microfluidic biochip technology, we have been able to separate these two processes and study the secretion pathways. Methods and results Adipogenic differentiation of human adipose-derived stem cells (ASCs) cultured in a microfluidic system was investigated under perfusion conditions with an adipogenic medium or an adipogenic medium supplemented with supernatant from differentiating ASCs (conditioned medium). Conditioned medium increased adipogenic differentiation compared to adipogenic medium with respect to accumulation of lipid-filled vacuoles and gene expression of key adipogenic markers (C/EBPα, C/EBPβ, C/EBPδ, PPARγ, LPL and adiponectin). The positive effects of conditioned medium were observed early in the differentiation process. Conclusions Using different cell densities and microfluidic perfusion cell cultures to suppress the effects of cell-released factors, we have demonstrated the significant role played by auto- or paracrine signaling in adipocyte differentiation. The cell-released factor(s) were shown to act in the recruitment phase of the differentiation process. PMID:23723991

  18. The role of paracrine and autocrine signaling in the early phase of adipogenic differentiation of adipose-derived stem cells.

    Directory of Open Access Journals (Sweden)

    Mette Hemmingsen

    Full Text Available INTRODUCTION: High cell density is known to enhance adipogenic differentiation of mesenchymal stem cells, suggesting secretion of signaling factors or cell-contact-mediated signaling. By employing microfluidic biochip technology, we have been able to separate these two processes and study the secretion pathways. METHODS AND RESULTS: Adipogenic differentiation of human adipose-derived stem cells (ASCs cultured in a microfluidic system was investigated under perfusion conditions with an adipogenic medium or an adipogenic medium supplemented with supernatant from differentiating ASCs (conditioned medium. Conditioned medium increased adipogenic differentiation compared to adipogenic medium with respect to accumulation of lipid-filled vacuoles and gene expression of key adipogenic markers (C/EBPα, C/EBPβ, C/EBPδ, PPARγ, LPL and adiponectin. The positive effects of conditioned medium were observed early in the differentiation process. CONCLUSIONS: Using different cell densities and microfluidic perfusion cell cultures to suppress the effects of cell-released factors, we have demonstrated the significant role played by auto- or paracrine signaling in adipocyte differentiation. The cell-released factor(s were shown to act in the recruitment phase of the differentiation process.

  19. Directed neuronal differentiation of human embryonic stem cells

    Directory of Open Access Journals (Sweden)

    Noggle Scott A

    2003-10-01

    Full Text Available Abstract Background We have developed a culture system for the efficient and directed differentiation of human embryonic stem cells (HESCs to neural precursors and neurons. HESC were maintained by manual passaging and were differentiated to a morphologically distinct OCT-4+/SSEA-4- monolayer cell type prior to the derivation of embryoid bodies. Embryoid bodies were grown in suspension in serum free conditions, in the presence of 50% conditioned medium from the human hepatocarcinoma cell line HepG2 (MedII. Results A neural precursor population was observed within HESC derived serum free embryoid bodies cultured in MedII conditioned medium, around 7–10 days after derivation. The neural precursors were organized into rosettes comprised of a central cavity surrounded by ring of cells, 4 to 8 cells in width. The central cells within rosettes were proliferating, as indicated by the presence of condensed mitotic chromosomes and by phosphoHistone H3 immunostaining. When plated and maintained in adherent culture, the rosettes of neural precursors were surrounded by large interwoven networks of neurites. Immunostaining demonstrated the expression of nestin in rosettes and associated non-neuronal cell types, and a radial expression of Map-2 in rosettes. Differentiated neurons expressed the markers Map-2 and Neurofilament H, and a subpopulation of the neurons expressed tyrosine hydroxylase, a marker for dopaminergic neurons. Conclusion This novel directed differentiation approach led to the efficient derivation of neuronal cultures from HESCs, including the differentiation of tyrosine hydroxylase expressing neurons. HESC were morphologically differentiated to a monolayer OCT-4+ cell type, which was used to derive embryoid bodies directly into serum free conditions. Exposure to the MedII conditioned medium enhanced the derivation of neural precursors, the first example of the effect of this conditioned medium on HESC.

  20. Chromogenic in situ hybridization compared with other approaches to evaluate HER2/neu status in breast carcinomas

    Energy Technology Data Exchange (ETDEWEB)

    Rosa, F.E.; Santos, R.M. [Departamento de Patologia, Hospital das Clínicas, Faculdade de Medicina, Universidade Estadual Paulista, Botucatu, SP (Brazil); Rogatto, S.R. [Departamento de Urologia, Faculdade de Medicina, Universidade Estadual Paulista, Botucatu, SP (Brazil); Hospital A.C. Camargo, CIPE, São Paulo, SP (Brazil); Domingues, M.A.C. [Departamento de Patologia, Faculdade de Medicina, Universidade Estadual Paulista, Botucatu, SP (Brazil)

    2013-03-19

    Human epidermal growth factor receptor 2 (HER2) has been evaluated in breast cancer patients to identify those most likely to benefit from herceptin-targeted therapy. HER2 amplification, detected in 20-30% of invasive breast tumors, is associated with reduced survival and metastasis. The most frequently used technique for evaluating HER2 protein status as a routine procedure is immunohistochemistry (IHC). HER2 copy number alterations have also been evaluated by fluorescence in situ hybridization (FISH) in moderate immunoexpression (IHC 2+) cases. An alternative procedure to evaluate gene amplification is chromogenic in situ hybridization (CISH), which has some advantages over FISH, including the correlation between HER2 status and morphological features. Other methodologies have also been used, such as silver-enhanced in situ hybridization (SISH) and quantitative real-time RT-PCR, to determine the number of HER2 gene copies and expression, respectively. Here we will present a short and comprehensive review of the current advances concerning HER2 evaluation in human breast cancer.

  1. Chromogenic in situ hybridization compared with other approaches to evaluate HER2/neu status in breast carcinomas

    Science.gov (United States)

    Rosa, F.E.; Santos, R.M.; Rogatto, S.R.; Domingues, M.A.C.

    2013-01-01

    Human epidermal growth factor receptor 2 (HER2) has been evaluated in breast cancer patients to identify those most likely to benefit from herceptin-targeted therapy. HER2 amplification, detected in 20-30% of invasive breast tumors, is associated with reduced survival and metastasis. The most frequently used technique for evaluating HER2 protein status as a routine procedure is immunohistochemistry (IHC). HER2 copy number alterations have also been evaluated by fluorescence in situ hybridization (FISH) in moderate immunoexpression (IHC 2+) cases. An alternative procedure to evaluate gene amplification is chromogenic in situ hybridization (CISH), which has some advantages over FISH, including the correlation between HER2 status and morphological features. Other methodologies have also been used, such as silver-enhanced in situ hybridization (SISH) and quantitative real-time RT-PCR, to determine the number of HER2 gene copies and expression, respectively. Here we will present a short and comprehensive review of the current advances concerning HER2 evaluation in human breast cancer. PMID:23558859

  2. Chromogenic in situ hybridization compared with other approaches to evaluate HER2/neu status in breast carcinomas

    International Nuclear Information System (INIS)

    Rosa, F.E.; Santos, R.M.; Rogatto, S.R.; Domingues, M.A.C.

    2013-01-01

    Human epidermal growth factor receptor 2 (HER2) has been evaluated in breast cancer patients to identify those most likely to benefit from herceptin-targeted therapy. HER2 amplification, detected in 20-30% of invasive breast tumors, is associated with reduced survival and metastasis. The most frequently used technique for evaluating HER2 protein status as a routine procedure is immunohistochemistry (IHC). HER2 copy number alterations have also been evaluated by fluorescence in situ hybridization (FISH) in moderate immunoexpression (IHC 2+) cases. An alternative procedure to evaluate gene amplification is chromogenic in situ hybridization (CISH), which has some advantages over FISH, including the correlation between HER2 status and morphological features. Other methodologies have also been used, such as silver-enhanced in situ hybridization (SISH) and quantitative real-time RT-PCR, to determine the number of HER2 gene copies and expression, respectively. Here we will present a short and comprehensive review of the current advances concerning HER2 evaluation in human breast cancer

  3. Magnetic resonance tomography of the orbit: First experiences with the paramagnetic contrast medium gadolinium-DTPA

    International Nuclear Information System (INIS)

    Markl, A.; Vogl, T.; Scheidhauer, K.; Riedel, K.G.; Oeckler, R.

    1986-01-01

    In 21 patients with orbital mass lesions MRI was performed before and after administration of paramagnetic contrast medium, gadolinium-DPTA. In comparison to the plain scan the differentiation of the tumorous tissue against the surrounding structures was improved after application of contrast medium despite a partially moderate increase in signal intensity. Especially highly vascular tumors and vessel diseases show a significant contrast enhancement. With increasing experience in larger number of patients a tissue differentiation seems to be possible. (orig.) [de

  4. Chromogenic in situ hybridization (CISH): a novel alternative in screening archival breast cancer tissue samples for HER-2/neu status.

    Science.gov (United States)

    Madrid, Manuelito A; Lo, Raymundo W

    2004-01-01

    Chromogenic in situ hybridization (CISH) is emerging as a practical, cost-effective, and valid alternative to fluorescent in situ hybridization in testing for gene alteration, especially in centers primarily working with immunohistochemistry (IHC). We assessed Her-2/neu alteration using CISH on formalin-fixed paraffin-embedded primary invasive ductal carcinoma tumors in which IHC (CB11 antibody) had previously been performed, and we compared the results with IHC. The 160 selected cases were equally stratified randomly into the four IHC categories (scores of 0, 1+, 2+, and 3+). We also compared age at diagnosis and tumor histologic grade with IHC and CISH Her-2/neu. We were able to perform and evaluate CISH successfully on all cases. The agreement between 3+ IHC and CISH-amplified cases as well as between all IHC and CISH Her-2/neu negative cases was 100%, and the concordance on all positive cases was 72.50%, with an overall agreement of 86.25%. All the discordant cases had 2+ IHC scores. Although we noted Her-2/neu positivity more in premenopausal women, the age at diagnosis was not significantly associated with IHC or CISH results. Similarly, although the small group of well-differentiated tumors was apparently Her-2/neu negative in both tests, no significant association was noted between any tumor histologic grade and either IHC or CISH results. CISH is easily integrated into routine testing in our laboratory. It is a necessary adjunct in determining the subset of non-amplified IHC-positive invasive tumors that will not benefit from trastuzumab therapy. Those cases with 2+ IHC results will be triaged and subjected to CISH. Her-2/neu testing should be done on all breast cancer cases regardless of age at presentation and tumor histologic grade.

  5. Chromogenic in situ hybridization (CISH): a novel alternative in screening archival breast cancer tissue samples for HER-2/neu status

    International Nuclear Information System (INIS)

    Madrid, Manuelito A; Lo, Raymundo W

    2004-01-01

    Chromogenic in situ hybridization (CISH) is emerging as a practical, cost-effective, and valid alternative to fluorescent in situ hybridization in testing for gene alteration, especially in centers primarily working with immunohistochemistry (IHC). We assessed Her-2/neu alteration using CISH on formalin-fixed paraffin-embedded primary invasive ductal carcinoma tumors in which IHC (CB11 antibody) had previously been performed, and we compared the results with IHC. The 160 selected cases were equally stratified randomly into the four IHC categories (scores of 0, 1+, 2+, and 3+). We also compared age at diagnosis and tumor histologic grade with IHC and CISH Her-2/neu. We were able to perform and evaluate CISH successfully on all cases. The agreement between 3+ IHC and CISH-amplified cases as well as between all IHC and CISH Her-2/neu negative cases was 100%, and the concordance on all positive cases was 72.50%, with an overall agreement of 86.25%. All the discordant cases had 2+ IHC scores. Although we noted Her-2/neu positivity more in premenopausal women, the age at diagnosis was not significantly associated with IHC or CISH results. Similarly, although the small group of well-differentiated tumors was apparently Her-2/neu negative in both tests, no significant association was noted between any tumor histologic grade and either IHC or CISH results. CISH is easily integrated into routine testing in our laboratory. It is a necessary adjunct in determining the subset of non-amplified IHC-positive invasive tumors that will not benefit from trastuzumab therapy. Those cases with 2+ IHC results will be triaged and subjected to CISH. Her-2/neu testing should be done on all breast cancer cases regardless of age at presentation and tumor histologic grade

  6. Application of photocatalytic cadmium sulfide nanoparticles to detection of enzymatic activities of glucose oxidase and glutathione reductase using oxidation of 3,3′,5,5′-tetramethylbenzidine

    Energy Technology Data Exchange (ETDEWEB)

    Grinyte, Ruta; Garai-Ibabe, Gaizka; Saa, Laura; Pavlov, Valeri, E-mail: vpavlov@cicbiomagune.es

    2015-06-30

    Highlights: • The light-powered nanosensor fabricated by enzymatic reactions was reported. • The sensor use energy of photons for oxidation of chromogenic enzymatic substrates. • Enzymatic assays for glucose oxidase and glutathione reductase were developed. - Abstract: It was found out that semiconductor CdS nanoparticles (NPs) are able to catalyze photooxidation of the well known chromogenic enzymatic substrate 3,3′,5,5′-tetramethylbenzidine (TMB) by oxygen. The photocatalytical oxidation of TMB does not require hydrogen peroxide and its rate is directly proportional to the quantity of CdS NPs produced in situ through the interaction of Cd{sup 2+} and S{sup 2−} ions in an aqueous medium. This phenomenon was applied to development of colorimetric sensitive assays for glucose oxidase and glutathione reductase based on enzymatic generation of CdS NPs acting as light-powered catalysts. Sensitivity of the developed chromogenic assays was of the same order of magnitude or even better than that of relevant fluorogenic assays. The present approach opens the possibility for the design of simple and sensitive colorimetric assays for a number of enzymes using inexpensive and available TMB as a universal chromogenic compound.

  7. Acceleration of astrocytic differentiation in neural stem cells surviving X-irradiation.

    Science.gov (United States)

    Ozeki, Ayumi; Suzuki, Keiji; Suzuki, Masatoshi; Ozawa, Hiroki; Yamashita, Shunichi

    2012-03-28

    Neural stem cells (NSCs) are highly susceptible to DNA double-strand breaks; however, little is known about the effects of radiation in cells surviving radiation. Although the nestin-positive NSCs predominantly became glial fibrillary acidic protein (GFAP)-positive in differentiation-permissive medium, little or no cells were GFAP positive in proliferation-permissive medium. We found that more than half of the cells surviving X-rays became GFAP positive in proliferation-permissive medium. Moreover, localized irradiation stimulated differentiation of cells outside the irradiated area. These results indicate for the first time that ionizing radiation is able to stimulate astrocyte-specific differentiation of surviving NSCs, whose process is mediated both by the direct activation of nuclear factor-κB and by the indirect bystander effect induced by X-irradiation.

  8. Spectral transfer functions of body waves propagating through a stratified medium. Part 1: Basic theory by means of matrix propagators

    International Nuclear Information System (INIS)

    Macia, R.; Correig, A.M.

    1987-01-01

    Seismic wave propagation is described by a second order differential equation for medium displacement. By Fourier transforming with respect to time and space, wave equation transforms into a system of first order linear differential equations for the Fourier transform of displacement and stress. This system of differential equations is solved by means of Matrix Propagator and applied to the propagation of body waves in stratified media. The matrix propagators corresponding to P-SV and SH waves in homogeneous medium are found as an intermediate step to obtain the spectral response of body waves propagating through a stratified medium with homogeneous layers. (author) 14 refs

  9. Chromogene properties of the betalains before gamma photons; Propiedades cromogenas de las betalainas ante fotones gamma

    Energy Technology Data Exchange (ETDEWEB)

    Rodriguez N, S.; Quintero M, C. L. [Universidad Autonoma de Zacatecas, Unidad Academica de Ciencias Quimicas, Programa de Quimica en Alimentos, Km. 0.5 Carretera a Guadalajara Ejido La Escondida, Zacatecas (Mexico); Vega C, H. R., E-mail: srneri@hotmail.co [Universidad Autonoma de Zacatecas, Unidad Academica de Ciencias Nucleares, Calle Cipres No. 10, Fracc. La Penuela, 98068 Zacatecas (Mexico)

    2010-09-15

    The coloration changes of four natural extracts in function of the absorbed dose produced by a gamma rays source of {sup 137}Cs have been studied. The natural extracts were obtained of tuna varieties that contain betalains that are natural pigments of some plants as the beet of where is derived its name. These also are found in abundant form in the fruits of some species of opuntia genus (tunas). The extracts were obtained by maceration, starting from beet and three tuna varieties that were stabilized to a p H of 5.5. The extracts were exposed to the gamma rays of a {sup 137}Cs source and the change in the coloration was observed by means of an ultra violet/visible spectrophotometer through of the absorption of the samples to photons of wave longitude 535 nm. The absorption was measured, to different time intervals. The relation between the absorbed dose in D{sub w} water and the chromogene properties of the pigment was established, with the intention of using it as possible dosemeter. (Author)

  10. Rho kinase inhibitor Y-27632 promotes the differentiation of human bone marrow mesenchymal stem cells into keratinocyte-like cells in xeno-free conditioned medium.

    Science.gov (United States)

    Li, Zhenzhen; Han, Shichao; Wang, Xingqin; Han, Fu; Zhu, Xiongxiang; Zheng, Zhao; Wang, Hongtao; Zhou, Qin; Wang, Yunchuan; Su, Linlin; Shi, Jihong; Tang, Chaowu; Hu, Dahai

    2015-03-11

    Bone marrow mesenchymal stem cells (BMSCs), which have the ability to self-renew and to differentiate into multiple cell types, have recently become a novel strategy for cell-based therapies. The differentiation of BMSCs into keratinocytes may be beneficial for patients with burns, disease, or trauma. However, the currently available cells are exposed to animal materials during their cultivation and induction. These xeno-contaminations severely limit their clinical outcomes. Previous studies have shown that the Rho kinase (ROCK) inhibitor Y-27632 can promote induction efficiency and regulate the self-renewal and differentiation of stem cells. In the present study, we attempted to establish a xeno-free system for the differentiation of BMSCs into keratinocytes and to investigate whether Y-27632 can facilitate this differentiation. BMSCs isolated from patients were cultured by using a xeno-free system and characterised by using flow cytometric analysis and adipogenic and osteogenic differentiation assays. Human primary keratinocytes were also isolated from patients. Then, the morphology, population doubling time, and β-galactosidase staining level of these cells were evaluated in the presence or absence of Y-27632 to determine the effects of Y-27632 on the state of the keratinocytes. Keratinocyte-like cells (KLCs) were detected at different time points by immunocytofluorescence analysis. Moreover, the efficiency of BMSC differentiation under different conditions was measured by quantitative real-time-polymerase chain reaction (RT-PCR) and Western blot analyses. The ROCK inhibitor Y-27632 promoted the proliferation and lifespan of human primary keratinocytes. In addition, we showed that keratinocyte-specific markers could be detected in BMSCs cultured in a xeno-free system using keratinocyte-conditioned medium (KCM) independent of the presence of Y-27632. However, the efficiency of the differentiation of BMSCs into KLCs was significantly higher in the presence of Y

  11. Medium Theory and Social Systems

    DEFF Research Database (Denmark)

    Tække, Jesper

      the  possibility  for  observation both of a social micro and a social macro level from a medium perspective. In the next  section  the paper  frames  the macro  level by  a  tentative  synthesis of  the medium  theory  and  the  sociological systems theory briefly describing a socio......-evolutionary process where new media alter  the societal capacity to handle complexity  in  time and space.  In  this section it becomes probable  that  by  means  of  different  media,  social  systems  give  different  possibilities  for  actual  social  performance.  In a way,  social  systems  themselves can be......  seen as medium  for  formation. Finally  the  paper  takes  the micro  level  perspective  by  applying  the  theory  to  newsgroups,  interpreting  them as self-organizing interactive systems giving a differentiated and diversified scope for social  inclusion.  ...

  12. Platelet lysate induces chondrogenic differentiation of umbilical cord-derived mesenchymal stem cells.

    Science.gov (United States)

    Hassan, Ghmkin; Bahjat, Mohammad; Kasem, Issam; Soukkarieh, Chadi; Aljamali, Majd

    2018-01-01

    Articular cartilage has a poor capacity for self-repair, and thus still presents a major challenge in orthopedics. Mesenchymal stem cells (MSCs) are multipotent stem cells with the potential to differentiate into chondrocytes in the presence of transforming growth factor beta (TGF-β). Platelet lysate (PL) contains a relatively large number of growth factors, including TGF-β, and has been shown to ameliorate cartilage repair. Here, we investigated the ability of PL to direct chondrogenic differentiation of MSCs along with other standard differentiation components in a pellet culture system. We isolated and expanded MSCs from human umbilical cords using a PL-supplemented medium and characterized the cells based on immunophenotype and potential for differentiation to adipocytes and osteocytes. We further cultured MSCs as pellets in a chondrogenic-differentiation medium supplemented with PL. After 21 days, the pellets were processed for histological analysis and stained with alician blue and acridine orange. The expression of SOX9 was investigated using RT-PCR. MSCs maintained their stemness characteristics in the PL-supplemented medium. However, the distribution of cells in the pellets cultured in the PL-supplemented chondrogenic differentiation medium had a greater similarity to cartilage tissue-derived chondrocytes than to the negative control. The intense alician blue staining indicated an increased production of mucopolysaccharides in the differentiated pellets, which also showed elevated expression of SOX9 . Our data suggest that MSCs could be differentiated to chondrocytes in the presence of PL and absence of exogenous TGF-β. Further research needs to be conducted to understand the exact role and potential of PL in chondrogenic differentiation and chondrocyte regeneration.

  13. Effects of Substrate and Co-Culture on Neural Progenitor Cell Differentiation

    Energy Technology Data Exchange (ETDEWEB)

    Jones, Erin Boote [Iowa State Univ., Ames, IA (United States)

    2008-01-01

    In recent years the study of stem and progenitor cells has moved to the forefront of research. Since the isolation of human hematopoietic stem cells in 1988 and the subsequent discovery of a self renewing population of multipotent cells in many tissues, many researchers have envisioned a better understanding of development and potential clinical usage in intractable diseases. Both these goals, however, depend on a solid understanding of the intracellular and extracellular forces that cause stem cells to differentiate to a specific cell fate. Many diseases of large scale cell loss have been suggested as candidates for stem cell based treatments. It is proposed that replacing the function of the damaged or defective cells by specific differentiation of stem or progenitor cells could treat the disease. Before cells can be directed to specific lineages, the mechanisms of differentiation must be better understood. Differentiation in vivo is an intensively complex system that is difficult to study. The goal of this research is to develop further understanding of the effects of soluble and extracellular matrix (ECM) cues on the differentiation of neural progenitor cells with the use of a simplified in vitro culture system. Specific research objectives are to study the differentiation of neural progenitor cells in response to astrocyte conditioned medium and protein substrate composition and concentration. In an effort to reveal the mechanism of the conditioned medium interaction, a test for the presence of a feedback loop between progenitor cells and astrocytes is presented along with an examination of conditioned medium storage temperature, which can reveal enzymatic dependencies. An examination of protein substrate composition and concentration will help to reveal the role of any ECM interactions on differentiation. This thesis is organized into a literature review covering recent advances in use of external modulators of differentiation such as surface coatings, co

  14. Differentiation of Human Dental Pulp Stem Cells into Dopaminergic Neuron-like Cells in Vitro.

    Science.gov (United States)

    Chun, So Young; Soker, Shay; Jang, Yu-Jin; Kwon, Tae Gyun; Yoo, Eun Sang

    2016-02-01

    We investigated the potential of human dental pulp stem cells (hDPSCs) to differentiate into dopaminergic neurons in vitro as an autologous stem cell source for Parkinson's disease treatment. The hDPSCs were expanded in knockout-embryonic stem cell (KO-ES) medium containing leukemia inhibitory factor (LIF) on gelatin-coated plates for 3-4 days. Then, the medium was replaced with KO-ES medium without LIF to allow the formation of the neurosphere for 4 days. The neurosphere was transferred into ITS medium, containing ITS (human insulin-transferrin-sodium) and fibronectin, to select for Nestin-positive cells for 6-8 days. The cells were then cultured in N-2 medium containing basic fibroblast growth factor (FGF), FGF-8b, sonic hedgehog-N, and ascorbic acid on poly-l-ornithine/fibronectin-coated plates to expand the Nestin-positive cells for up to 2 weeks. Finally, the cells were transferred into N-2/ascorbic acid medium to allow for their differentiation into dopaminergic neurons for 10-15 days. The differentiation stages were confirmed by morphological, immunocytochemical, flow cytometric, real-time PCR, and ELISA analyses. The expressions of mesenchymal stem cell markers were observed at the early stages. The expressions of early neuronal markers were maintained throughout the differentiation stages. The mature neural markers showed increased expression from stage 3 onwards. The percentage of cells positive for tyrosine hydroxylase was 14.49%, and the amount was 0.526 ± 0.033 ng/mL at the last stage. hDPSCs can differentiate into dopaminergic neural cells under experimental cell differentiation conditions, showing potential as an autologous cell source for the treatment of Parkinson's disease.

  15. Differential Effect of Medium on the Ratio of ICM/TE of Bovine Embryos in a Co-culture System

    Directory of Open Access Journals (Sweden)

    Mohsen Forouzanfar

    2010-01-01

    Full Text Available Background: This study was undertaken to investigate the efficiency of two differentembryo somatic cell co-culture conditions, tissue culture medium 199 (TCM199–vero cellsand Menezo B2 (B2-vero cells, for the in vitro developmental quantity and quality of bovineembryos.Materials and Methods: Bovine oocytes were allowed to mature and subsequently undergofertilization in vitro. Their presumptive zygotes were cultured in either TCM199 or B2 culturemedia in conjunction with vero cells for up to nine days. The culture media were refreshedevery two days and the proportion of embryos that cleaved and further developed to themorula and blastocyst (early, expand and hatched stages were recorded. Hatched blastocystsunderwent differential staining in order to determine the numbers of inner cell mass (ICMand tropho ectoderm (TE and total cell number (TCN.Results: Of the two groups, no significant difference was seen between the proportions ofthe presumptive zygotes cleaved, those which developed to 8-16 cells, morula and reacheddays 7or 8 blastocyst stage or hatched. However, the values for TCN and TE of the TCM199-vero embryos were significantly greater than those of B2-vero embryos. The values for ICM/TCN and ICM/TE were significantly greater in the B2-vero group versus the TCM199-verogroup.Conclusion: Both TCM199 and B2 culture media in conjunction with vero cells were ofthe same efficiency when used for in vitro development of bovine presumptive zygotes.However, TCM199 was superior in providing embryos with more embryo cell numbers,whereas B2 medium was superior in providing embryos with greater ICM/TE and ICM/TCN ratios.

  16. Probing the nuclear medium with the K+ meson

    International Nuclear Information System (INIS)

    Chrien, R.E.

    1995-01-01

    Elastic differential cross sections for K + mesons scattered from targets of carbon and 6 Li have been measured at an incident momentum of 715 MeV/c. The ratios of scattering cross sections from these targets are not predicted by theory, and are consistent with earlier suggestions that the K + -nucleon interaction is modified in the nuclear medium

  17. The Role of Branding in Small and Medium-Sized Enterprises

    Directory of Open Access Journals (Sweden)

    Franc Vidic

    2013-12-01

    Full Text Available The purpose of this article is to show the relationship between branding and brand management in small and medium-sized enterprises (SMEs. Traditionally, branding was associated with large and global corporations. However, we often forget that small and medium-sized enterprises also deal with their own names (brands in their own way. The study identified four types of businesses, regardless of their association with brands. We named these four types, as follows: Ignorant; User; Low-Cost Producer; and Differentiation Producer. If the first two types (i.e. Ignorant and User differ primarily in the extent to which they use simple branding activities, and are used mainly in the local market where the enterprises tend to operate, we found that the last two types (i.e. low-cost producers and differentiation producers design their branding strategies in accordance with their generic strategies and mode of growth.

  18. Chromogenic in situ hybridization for Her-2/neu-oncogene in breast cancer: comparison of a new dual-colour chromogenic in situ hybridization with immunohistochemistry and fluorescence in situ hybridization.

    Science.gov (United States)

    Mayr, Doris; Heim, Sibylle; Weyrauch, Kerstin; Zeindl-Eberhart, Evelyn; Kunz, Anne; Engel, Jutta; Kirchner, Thomas

    2009-12-01

    Her-2/neu testing is used as a marker for Herceptin therapy. The aim was to investigate new dual-colour chromogenic in situ hybridization (CISH), in a large number of breast carcinomas (n = 205) with DNA-specific dual-colour probes (ZytoVision, Bremerhaven, Germany) and to compare the results with immunohistochemistry (n = 205) and fluorescence in situ hybridization (FISH) (n = 129). Paraffin-embedded tissue of 205 patients was used. After immunohistochemistry with a focus on immunohistochemically uncertain cases, Her-2/neu amplification using dual-colour CISH (ZytoVision) was analysed. Validation by FISH was performed. The results were: immunohistochemistry, 27.8% with strong expression, 53.7% with uncertain overexpression and 18.5% with no expression; FISH, 25.6% amplified and 74.4% negative; CISH, 35.6% amplified, 62.9% negative and 1.5% not evaluable. Comparison of immunohistochemistry with CISH: CISH negative in 100% with immunohistochemistry 0/1+, amplified in 82.5% with immunohistochemistry 3+; 5.9% contradictory results: 4.4% immunohistochemistry 3+ and negative by CISH, 1.5% negative in immunohistochemistry but amplified by CISH; FISH (129 cases), 8.5% contradictory results to immunohistochemistry, 6.2% immunohistochemistry 3+ and negative by FISH, 2.3% negative by immunohistochemistry and amplified by FISH; comparison of CISH and FISH, 94.6% same results, 3.9% different ones, 1.6% CISH not analysable. CISH, using dual-colour probes (ZytoVision) is as good as FISH for Her-2/neu analysis. The few discrepant results are likely to be caused by polysomy or tumour heterogeneity.

  19. Chromogenic in situ hybridization compared with other approaches to evaluate HER2/neu status in breast carcinomas

    Directory of Open Access Journals (Sweden)

    F.E. Rosa

    Full Text Available Human epidermal growth factor receptor 2 (HER2 has been evaluated in breast cancer patients to identify those most likely to benefit from herceptin-targeted therapy. HER2 amplification, detected in 20-30% of invasive breast tumors, is associated with reduced survival and metastasis. The most frequently used technique for evaluating HER2 protein status as a routine procedure is immunohistochemistry (IHC. HER2 copy number alterations have also been evaluated by fluorescence in situ hybridization (FISH in moderate immunoexpression (IHC 2+ cases. An alternative procedure to evaluate gene amplification is chromogenic in situ hybridization (CISH, which has some advantages over FISH, including the correlation between HER2 status and morphological features. Other methodologies have also been used, such as silver-enhanced in situ hybridization (SISH and quantitative real-time RT-PCR, to determine the number of HER2 gene copies and expression, respectively. Here we will present a short and comprehensive review of the current advances concerning HER2 evaluation in human breast cancer.

  20. Chromogenic in situ hybridization compared with other approaches to evaluate HER2/neu status in breast carcinomas

    Directory of Open Access Journals (Sweden)

    F.E. Rosa

    2013-03-01

    Full Text Available Human epidermal growth factor receptor 2 (HER2 has been evaluated in breast cancer patients to identify those most likely to benefit from herceptin-targeted therapy. HER2 amplification, detected in 20-30% of invasive breast tumors, is associated with reduced survival and metastasis. The most frequently used technique for evaluating HER2 protein status as a routine procedure is immunohistochemistry (IHC. HER2 copy number alterations have also been evaluated by fluorescence in situ hybridization (FISH in moderate immunoexpression (IHC 2+ cases. An alternative procedure to evaluate gene amplification is chromogenic in situ hybridization (CISH, which has some advantages over FISH, including the correlation between HER2 status and morphological features. Other methodologies have also been used, such as silver-enhanced in situ hybridization (SISH and quantitative real-time RT-PCR, to determine the number of HER2 gene copies and expression, respectively. Here we will present a short and comprehensive review of the current advances concerning HER2 evaluation in human breast cancer.

  1. Probing the mechanisms of drug release from amorphous solid dispersions in medium-soluble and medium-insoluble carriers.

    Science.gov (United States)

    Sun, Dajun D; Lee, Ping I

    2015-08-10

    The objective of the current study is to mechanistically differentiate the dissolution and supersaturation behaviors of amorphous drugs from amorphous solid dispersions (ASDs) based on medium-soluble versus medium-insoluble carriers under nonsink dissolution conditions through a direct head-to-head comparison. ASDs of indomethacin (IND) were prepared in several polymers which exhibit different solubility behaviors in acidic (pH1.2) and basic (pH7.4) dissolution media. The selected polymers range from water-soluble (e.g., PVP and Soluplus) and water-insoluble (e.g., ethylcellulose and Eudragit RL PO) to those only soluble in an acidic or basic dissolution medium (e.g., Eudragit E100, Eudragit L100, and HPMCAS). At 20wt.% drug loading, DSC and powder XRD analysis confirmed that the majority of incorporated IND was present in an amorphous state. Our nonsink dissolution results confirm that whether the carrier matrix is medium soluble determines the release mechanism of amorphous drugs from ASD systems which has a direct impact on the rate of supersaturation generation, thus in turn affecting the evolution of supersaturation in amorphous systems. For example, under nonsink dissolution conditions, the release of amorphous IND from medium-soluble carriers is governed by a dissolution-controlled mechanism leading to an initial surge of supersaturation followed by a sharp decline in drug concentration due to rapid nucleation and crystallization. In contrast, the dissolution of IND ASD from medium-insoluble carriers is more gradual as drug release is regulated by a diffusion-controlled mechanism by which drug supersaturation is built up gradually and sustained over an extended period of time without any apparent decline. Since several tested carrier polymers can be switched from soluble to insoluble by simply changing the pH of the dissolution medium, the results obtained here provide unequivocal evidence of the proposed transition of kinetic solubility profiles from the

  2. Shoot Differentiation in Callus Cultures of Datura Innoxia

    DEFF Research Database (Denmark)

    Engvild, Kjeld Christensen

    1973-01-01

    promoted shoot differentiation. Gibberellic acid inhibited shoot formation weakly, but inhibited proper leaf blade formation. Root differentiation was rare. The callus cultures of Datura innoxia grew rapidly (100-fold in 4 weeks) on a slightly modified Murashige and Skoog medium (0.5 mg/l thiamin · HCl, p...

  3. Evaluation of insulin medium or chondrogenic medium on proliferation and chondrogenesis of ATDC5 cells.

    Science.gov (United States)

    Yao, Yongchang; Zhai, Zhichen; Wang, Yingjun

    2014-01-01

    The ATDC5 cell line is regarded as an excellent cell model for chondrogenesis. In most studies with ATDC5 cells, insulin medium (IM) was used to induce chondrogenesis while chondrogenic medium (CM), which was usually applied in chondrogenesis of mesenchymal stem cells (MSCs), was rarely used for ATDC5 cells. This study was mainly designed to investigate the effect of IM, CM, and growth medium (GM) on chondrogenesis of ATDC5 cells. ATDC5 cells were, respectively, cultured in IM, CM, and GM for a certain time. Then the proliferation and the chondrogenesis progress of cells in these groups were analyzed. Compared with CM and GM, IM promoted the proliferation of cells significantly. CM was effective for enhancement of cartilage specific markers, while IM induced the cells to express endochondral ossification related genes. Although GAG deposition per cell in CM group was significantly higher than that in IM and GM groups, the total GAG contents in IM group were the most. This study demonstrated that CM focused on induction of chondrogenic differentiation while IM was in favor of promoting proliferation and expression of endochondral ossification related genes. Combinational use of these two media would be more beneficial to bone/cartilage repair.

  4. Evaluation of Insulin Medium or Chondrogenic Medium on Proliferation and Chondrogenesis of ATDC5 Cells

    Directory of Open Access Journals (Sweden)

    Yongchang Yao

    2014-01-01

    Full Text Available Background. The ATDC5 cell line is regarded as an excellent cell model for chondrogenesis. In most studies with ATDC5 cells, insulin medium (IM was used to induce chondrogenesis while chondrogenic medium (CM, which was usually applied in chondrogenesis of mesenchymal stem cells (MSCs, was rarely used for ATDC5 cells. This study was mainly designed to investigate the effect of IM, CM, and growth medium (GM on chondrogenesis of ATDC5 cells. Methods. ATDC5 cells were, respectively, cultured in IM, CM, and GM for a certain time. Then the proliferation and the chondrogenesis progress of cells in these groups were analyzed. Results. Compared with CM and GM, IM promoted the proliferation of cells significantly. CM was effective for enhancement of cartilage specific markers, while IM induced the cells to express endochondral ossification related genes. Although GAG deposition per cell in CM group was significantly higher than that in IM and GM groups, the total GAG contents in IM group were the most. Conclusion. This study demonstrated that CM focused on induction of chondrogenic differentiation while IM was in favor of promoting proliferation and expression of endochondral ossification related genes. Combinational use of these two media would be more beneficial to bone/cartilage repair.

  5. Role of chromogenic in situ hybridization (CISH) in the evaluation of HER2 status in breast carcinoma: comparison with immunohistochemistry and FISH.

    Science.gov (United States)

    Li-Ning-T, Elsa; Ronchetti, Ruben; Torres-Cabala, Carlos; Merino, Maria J

    2005-10-01

    We report our experience with Chromogenic in Situ Hybridization (CISH) for the evaluation of HER2 amplification on 55 cases of formalin-fixed, paraffin-embedded invasive breast carcinomas of different histology. All the results were corrected for chromosome 17 aneusomy and compared with immunohistochemistry (IHC); a subset of cases was compared to FISH. Thirty-one of 32 cases in which FISH and CISH were performed yielded the same results. CISH and IHC showed a good concordance in the 0/1+ and 3+ category, while a poor agreement with weakly protein overexpression was confirmed. Chromosome 17 analysis was necessary in cases with a low number of HER2 gene copies. CISH is a useful tool to evaluate breast cancer HER2 status that can be easily implemented in a laboratory of surgical pathology.

  6. In vitro differentiation of human umbilical cord blood mesenchymal ...

    African Journals Online (AJOL)

    Mesenchymal stem cells (MSCs) were isolated by gradient density centrifugation from umbilical cord blood. Spindle-shaped adherent cells were permitted to grow to 70% confluence in primary culture media which was reached by day 12. Induction of differentiation started by culturing cells with differentiation medium ...

  7. Effects of Co-Culture Media on Hepatic Differentiation of hiPSC with or without HUVEC Co-Culture.

    Science.gov (United States)

    Freyer, Nora; Greuel, Selina; Knöspel, Fanny; Strahl, Nadja; Amini, Leila; Jacobs, Frank; Monshouwer, Mario; Zeilinger, Katrin

    2017-08-07

    The derivation of hepatocytes from human induced pluripotent stem cells (hiPSC) is of great interest for applications in pharmacological research. However, full maturation of hiPSC-derived hepatocytes has not yet been achieved in vitro. To improve hepatic differentiation, co-cultivation of hiPSC with human umbilical vein endothelial cells (HUVEC) during hepatic differentiation was investigated in this study. In the first step, different culture media variations based on hepatocyte culture medium (HCM) were tested in HUVEC mono-cultures to establish a suitable culture medium for co-culture experiments. Based on the results, two media variants were selected to differentiate hiPSC-derived definitive endodermal (DE) cells into mature hepatocytes with or without HUVEC addition. DE cells differentiated in mono-cultures in the presence of those media variants showed a significant increase ( p < 0.05) in secretion of α-fetoprotein and in activities of cytochrome P450 (CYP) isoenzymes CYP2B6 and CYP3A4 as compared with cells differentiated in unmodified HCM used as control. Co-cultivation with HUVEC did not further improve the differentiation outcome. Thus, it can be concluded that the effect of the used medium outweighed the effect of HUVEC co-culture, emphasizing the importance of the culture medium composition for hiPSC differentiation.

  8. Real group velocity in a medium with dissipation

    International Nuclear Information System (INIS)

    Muschietti, L.; Dum, C.T.

    1993-01-01

    When a medium is dissipative, the classic expression for the group velocity, dω/dk, is complex with an imaginary part often being far from negligible. To clarify the role of this imaginary term, the motion of a wave packet in a dissipative, homogeneous medium is examined. The integral representation of the packet is analyzed by means of a saddle-point method. It is shown that in a moving frame attached to its maximum the packet looks self-similar. A Gaussian packet keeps its Gaussian identity, as is familiar for the case of a nondissipative medium. However, the central wave number of the packet slowly changes because of a differential damping among the Fourier components: Im(dω/dk)=dγ/dk≠0, where ω≡ω r +iγ. The packet height can be computed self-consistently as integrated damping (or growth). The real group velocity becomes a time-dependent combination of Re(dω/dk) and Im(dω/dk). Only where the medium is both homogeneous and loss free, does the group velocity remain constant. Simple ''ray-tracing equations'' are derived to follow the packet centers in coordinate and Fourier spaces. The analysis is illustrated with a comparison to geometric optics, and by two applications: the case of a medium with some resonant damping (or growth) and the propagation of whistler waves in a collisional plasma

  9. Osteogenic differentiation of immature osteoblasts: Interplay of cell culture media and supplements.

    Science.gov (United States)

    Brauer, A; Pohlemann, T; Metzger, W

    2016-01-01

    Differentiation of immature osteoblasts to mature osteoblasts in vitro initially was induced by supplementing the medium with β-gylcerophosphate and dexamethasone. Later, ascorbic acid, vitamin D3, vitamin K3 and TGFβ1 were used in varying concentrations as supplements to generate a mature osteoblast phenotype. We tested the effects of several combinations of cell culture media, seeding protocols and osteogenic supplements on osteogenic differentiation of human primary osteoblasts. Osteogenic differentiation was analyzed by staining alkaline phosphatase (ALP) with 5-bromo-4-chloro-3-indolyl-phosphate/nitro blue tetrazolium (BCIP/NBT) and by von Kossa staining of deposited calcium phosphate. The combinations of culture media and supplements significantly influenced osteogenic differentiation, but the seeding protocol did not. Staining of ALP and calcium phosphate could be achieved only if our own mix of osteogenic supplements was used in combination with Dulbecco's modified Eagle medium or if a commercial mix of osteogenic supplements was used in combination with osteoblast growth medium. Especially for von Kossa, we observed great variations in the staining intensity. Because osteogenic differentiation is a complex process, the origin of the osteoblasts, cell culture media and osteogenic supplements should be established by preliminary experiments to achieve optimal differentiation. Staining of ALP or deposited calcium phosphate should be supplemented with qRT-PCR studies to learn more about the influence of specific supplements on osteogenic markers.

  10. Comparison of different methods for erythroid differentiation in the K562 cell line.

    Science.gov (United States)

    Shariati, Laleh; Modaress, Mehran; Khanahmad, Hossein; Hejazi, Zahra; Tabatabaiefar, Mohammad Amin; Salehi, Mansoor; Modarressi, Mohammad Hossein

    2016-08-01

    To compare methods for erythroid differentiation of K562 cells that will be promising in the treatment of beta-thalassemia by inducing γ-globin synthesis. Cells were treated separately with: RPMI 1640 medium without glutamine, RPMI 1640 medium without glutamine supplemented with 1 mM sodium butyrate, RPMI 1640 medium supplemented with 1 mM sodium butyrate, 25 µg cisplatin/ml, 0.1 µg cytosine arabinoside/ml. The highest differentiation (84 %) with minimum toxicity was obtained with cisplatin at 15 µg /ml. Real-time RT-PCR showed that expression of the γ-globin gene was significantly higher in the cells differentiated with cisplatin compared to undifferentiated cells (P < 0.001). Cisplatin is useful in the experimental therapy of ß-globin gene defects and can be considered for examining the basic mechanism of γ-reactivation.

  11. Ti nanorod arrays with a medium density significantly promote osteogenesis and osteointegration

    Science.gov (United States)

    Ning, Chengyun; Wang, Shuangying; Zhu, Ye; Zhong, Meiling; Lin, Xi; Zhang, Yu; Tan, Guoxin; Li, Mei; Yin, Zhaoyi; Yu, Peng; Wang, Xiaolan; Li, Ying; He, Tianrui; Chen, Wei; Wang, Yingjun; Mao, Chuanbin

    2016-01-01

    Ti implants are good candidates in bone repair. However, how to promote bone formation on their surface and their consequent perfect integration with the surrounding tissue is still a challenge. To overcome such challenge, we propose to form Ti nanorods on their surface to promote the new bone formation around the implants. Here Ti nanorod arrays (TNrs) with different densities were produced on pure Ti surfaces using an anodizing method. The influence of TNr density on the protein adsorption as well as on the adhesion, proliferation, and osteogenic differentiation of MC3T3-E1 pre-osteoblastic cells were assessed. The TNrs were also implanted into the bone defects in rabbits to test their application in promoting bone formation and osteointegration at the implant-bone interface. TNrs with the medium density were found to show the best capability in promoting the protein adsorption from surrounding medium, which in turn efficiently enhanced osteogenic differentiation in vitro and osteointegration in vivo. Our work suggests that growing TNrs with a medium density on the surface of traditional Ti implants is an efficient and facile method for promoting bone formation and osteointegration in bone repair.

  12. Changes in the chromogene properties of the betalaine; Cambios en las propiedades cromogenas de las betalainas inducidos por radiacion gamma

    Energy Technology Data Exchange (ETDEWEB)

    Rodriguez N, S.; Pinedo S, A.; Amador V, P.; Chacon R, A.; Arcos P, A.; Vega C, H.R. [Unidad Academica de Ciencias Nucleares, C. Cipres 10, Fracc. La Penuela, 98068, Zacatecas, Zac. (Mexico)

    2005-07-01

    The changes of coloration of four natural extracts were determined in function of the absorbed dose taken place by a source of rays gamma of Cs-137. The used natural extracts contain betalaine that are natural pigments of some plants as the beet root, of there their name. They are also in abundant form in the fruits (tunas) of some species of the Opuntia gender. The extracts were obtained by maceration, starting from beet root and three tuna varieties, and they were stabilized to pH 5.5 the change of coloration you determines in a spectrophotometer ultraviolet/visible by means of the absorbance from the samples to photons of 475, 535 and 600 nm of wave longitude. The absorbance was measured, to different intervals of time. The relationship settled down between the absorbed dose and the chromogene properties of the pigment, with the intention of using it as possible dosemeter. (Author)

  13. The Effects of Viscous Dissipation on Convection in a Porus Medium

    Directory of Open Access Journals (Sweden)

    T Raja Rani

    2017-05-01

    Full Text Available The aim of this paper is to study of the effects of variable physical properties and viscous dissipation on a free convective flow over a vertical plate with a variable temperature embedded in a porous medium. We study the effects of varying physical properties on heat transfer and on flow when the medium is filled with some commonly used experimental fluids, in particular, Glycerin, Water and Methyl chloride (a commonly refrigerant. A similarity transformation technique is used to reduce the partial differential equations governing the flow. The resulting system of non-linear coupled ordinary differential equations is solved numerically with appropriate boundary conditions using the Runge-Kutta-Gill method coupled with a shooting technique. Using this approach, a study is conducted on both hot and cold plates and results presented using a combination of graphical illustrations and tables of the effect of changing a variety of physical parameters, in particular, the temperature and viscosity of the fluid.

  14. Thermal non-equilibrium in porous medium adjacent to vertical plate: ANN approach

    Science.gov (United States)

    Ahmed, N. J. Salman; Ahamed, K. S. Nazim; Al-Rashed, Abdullah A. A. A.; Kamangar, Sarfaraz; Athani, Abdulgaphur

    2018-05-01

    Thermal non-equilibrium in porous medium is a condition that refers to temperature discrepancy in solid matrix and fluid of porous medium. This type of flow is complex flow requiring complex set of partial differential equations that govern the flow behavior. The current work is undertaken to predict the thermal non-equilibrium behavior of porous medium adjacent to vertical plate using artificial neural network. A set of neurons in 3 layers are trained to predict the heat transfer characteristics. It is found that the thermal non-equilibrium heat transfer behavior in terms of Nusselt number of fluid as well as solid phase can be predicted accurately by using well-trained neural network.

  15. On the viscous dissipation modeling of thermal fluid flow in a porous medium

    KAUST Repository

    Salama, Amgad; El-Amin, Mohamed; Abbas, Ibrahim A A; Sun, Shuyu

    2011-01-01

    wall that is immersed in the porous medium and is kept at constant higher temperature. The boundary layer approximations were used to simplify the set of the governing, nonlinear partial differential equations, which were then non

  16. Antibacterial activities of coagulase-negative staphylococci from bovine teat apex skin and their inhibitory effect on mastitis-related pathogens.

    Science.gov (United States)

    Braem, G; Stijlemans, B; Van Haken, W; De Vliegher, S; De Vuyst, L; Leroy, F

    2014-05-01

    To explore antibacterial activities of coagulase-negative staphylococci (CoNS) from teat apices of dairy cows towards mastitis-causing pathogens. Of 254 CoNS, 38 displayed bacteriocin-like activity after a first screening. Seven of these strains displayed activity against at least one mastitis-related pathogen (Streptococcus uberis, Streptococcus dysgalactiae and Staphylococcus aureus). Staphylococcus chromogenes L217 displayed the strongest inhibitory effect, being active against all tested mastitis-related pathogens and most tested CoNS. Based on cation exchange and reversed-phase chromatography, in addition to N-terminal Edman degradation and PCR, the antibacterial peptide was identified as a nukacin-type bacteriocin and named nukacin L217. Although staphylococcal bacteriocins are generally found in the cell-free supernatants of liquid cultures, Staph. chromogenes L217 only led to detectable activity when grown on agar medium. Bacteriocin-like activities are not uncommon among CoNS from teat apices and may inhibit mastitis-causing pathogens, as found for nukacin L217 production by Staph. chromogenes L217. Nukacin L217 is the first identified bacteriocin of the species Staph. chromogenes and displays unusual production kinetics, that is, requiring surface growth of its producer. The fact that nukacins are produced by different CoNS species suggests a role in the teat skin ecosystem. © 2014 The Society for Applied Microbiology.

  17. Bioactive glass ions as strong enhancers of osteogenic differentiation in human adipose stem cells.

    Science.gov (United States)

    Ojansivu, Miina; Vanhatupa, Sari; Björkvik, Leena; Häkkänen, Heikki; Kellomäki, Minna; Autio, Reija; Ihalainen, Janne A; Hupa, Leena; Miettinen, Susanna

    2015-07-01

    Bioactive glasses are known for their ability to induce osteogenic differentiation of stem cells. To elucidate the mechanism of the osteoinductivity in more detail, we studied whether ionic extracts prepared from a commercial glass S53P4 and from three experimental glasses (2-06, 1-06 and 3-06) are alone sufficient to induce osteogenic differentiation of human adipose stem cells. Cells were cultured using basic medium or osteogenic medium as extract basis. Our results indicate that cells stay viable in all the glass extracts for the whole culturing period, 14 days. At 14 days the mineralization in osteogenic medium extracts was excessive compared to the control. Parallel to the increased mineralization we observed a decrease in the cell amount. Raman and Laser Induced Breakdown Spectroscopy analyses confirmed that the mineral consisted of calcium phosphates. Consistently, the osteogenic medium extracts also increased osteocalcin production and collagen Type-I accumulation in the extracellular matrix at 13 days. Of the four osteogenic medium extracts, 2-06 and 3-06 induced the best responses of osteogenesis. However, regardless of the enhanced mineral formation, alkaline phosphatase activity was not promoted by the extracts. The osteogenic medium extracts could potentially provide a fast and effective way to differentiate human adipose stem cells in vitro. Copyright © 2015 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

  18. Direct identification and recognition of yeast species from clinical material by using albicans ID and CHROMagar Candida plates.

    Science.gov (United States)

    Baumgartner, C; Freydiere, A M; Gille, Y

    1996-02-01

    Two chromogenic media, Albicans ID and CHROMagar Candida agar plates, were compared with a reference medium, Sabouraud-chloramphenicol agar, and standard methods for the identification of yeast species. This study involved 951 clinical specimens. The detection rates for the two chromogenic media for polymicrobial specimens were 20% higher than that for the Sabouraud-chloramphenicol agar plates. The rates of identification of Candida albicans for Albicans ID and CHROMagar Candida agar plates were, respectively, 37.0 and 6.0% after 24 h of incubation and 93.6 and 92.2% after 72 h of incubation, with specificities of 99.8 and 100%. Furthermore, CHROMagar Candida plates identified 13 of 14 Candida tropicalis and 9 of 12 Candida krusei strains after 48 h of incubation.

  19. Study of the sensitivity of the radiation transport problem in a scattering medium

    International Nuclear Information System (INIS)

    Nunes, Rogerio Chaffin

    2002-03-01

    In this work, the system of differential equations obtained by the angular approach of the two-dimensional transport equation by the discrete ordinates method is solved through the formulation of finite elements with the objective of investigating the sensitivity of the outgoing flux of radiation with the incoming flux and the properties of absorption and scattering of the medium. The variational formulation for the system of differential equations of second order with the generalized boundary conditions of Neumann (third type) allows an easy implementation of the method of the finite elements with triangular mesh and approximation space of first order. The geometry chosen for the simulations is a circle with a non homogeneous circular form in its interior. The mapping of Dirichlet-Neumann is studied through various simulations involving the incoming flux, the outgoing flux and the properties of the medium. (author)

  20. Assessment of topoisomerase II-alpha gene status by dual color chromogenic in situ hybridization in a set of Iraqi patients with invasive breast carcinoma

    Directory of Open Access Journals (Sweden)

    Rasha Abd Alraouf Neama

    2017-01-01

    Full Text Available Background: The human epidermal growth factor receptor 2(HER2 proto-oncogene is overexpressed or amplified in approximately 15%–25% of invasive breast cancers. Approximately 35% of HER2-amplified breast cancers have coamplification of the topoisomerase II-alpha (TOP2A gene encoding an enzyme that is a major target of anthracyclines. Hence, the determination of genetic alteration (amplification or deletion of both genes is considered as an important predictive factor that determines the response of breast cancer patients to treatment. The aims of this study are to determinate TOP2A status gene amplification in a set of Iraqi patients with breast cancer that have had an equivocal (2+ and positive HER2/neu by immunohistochemistry (IHC and to compare the results with estrogen receptor (ER and progesterone receptor (PR and HER2/neu status. Patients and Methods: A cross-sectional prospective study done on 53 patients with invasive breast carcinoma. Twenty-six out of total 53 cases were positive HER2/neu (3+, the remaining 27 equivocal HER2-IHC (2+ cases reanalyzed using dual-color chromogenic in situ hybridization (ZytoVision probe kit for further identification of HER2/neu gene amplification. Using chromogenic in situ hybridization (CISH, TOP2A gene status determination was done for all cases. Results: There is a direct significant correlation between TOP2A gene amplification and HER2/neu positivity, P < 0.05 in that 15 (39.4% out of 38 positive HER2/neu cases were associated with topoisomerase gene amplification. Regarding relation of topoisomerase gene to hormone receptor status (ER and PR, there was a significant negative relationship between the gene and ER receptor status. The higher level of gene amplification was noticed in ER and PR negative cases in about 13 (43.3% and 14 (48.2% for ER and PR, respectively. Conclusion: TOP2A gene status has a significantly positive correlation with HER2/neu status while it has a significantly negative

  1. Effect of all-trans retinoic acid on the proliferation and differentiation of brain tumor stem cells

    Directory of Open Access Journals (Sweden)

    Niu Chao

    2010-08-01

    Full Text Available Abstract Objective To investigate the effect of all-trans retinoic acid(ATRA on the proliferation and differentiation of brain tumor stem cells(BTSCs in vitro. Methods Limiting dilution and clonogenic assay were used to isolate and screen BTSCs from the fresh specimen of human brain glioblastoma. The obtained BTSCs, which were cultured in serum-free medium, were classified into four groups in accordance with the composition of the different treatments. The proliferation of the BTSCs was evaluated by MTT assay. The BTSCs were induced to differentiate in serum-containing medium, and classified into the ATRA group and control group. On the 10th day of induction, the expressions of CD133 and glial fibrillary acidic protein (GFAP in the differentiated BTSCs were detected by immunofluorescence. The differentiated BTSCs were cultured in serum-free medium, the percentage and the time required for formation of brain tumor spheres (BTS were observed. Results BTSCs obtained by limiting dilution were all identified as CD133-positive by immunofluorescence. In serum-free medium, the proliferation of BTSCs in the ATRA group was observed significantly faster than that in the control group, but slower than that in the growth factor group and ATRA/growth factor group, and the size of the BTS in the ATRA group was smaller than that in the latter two groups(P P P P Conclusion ATRA can promote the proliferation and induce the differentiation of BTSCs, but the differentiation is incomplete, terminal differentiation cannot be achieved and BTSs can be formed again.

  2. Chromogenic in situ Hybridization Compared with Real time Quantitative Polymerase Chain Reaction to Evaluate HER2/neu Status in Breast Cancer.

    Science.gov (United States)

    Ayatollahi, Hossein; Fani, Azar; Ghayoor Karimiani, Ehsan; Homaee, Fateme; Shajiei, Arezoo; Sheikh, Maryam; Shakeri, Sepideh; Shams, Seyyede Fatemeh

    2017-01-01

    The assessment of human epidermal growth factor receptor 2 (HER2) status has become of great importance in the diagnosis of breast cancer. The aim of this study was to investigate the diagnostic value of quantitative Polymerase Chain Reaction (qPCR) and Chromogenic In Situ Hybridization (CISH) to assess HER2 status of biopsy specimens. To elucidate the status of HER2 gene amplification, biopsies of breast carcinoma from 120 patients with 2+ IHC status were analyzed by qPCR and CISH. The results of the two experiments were compared, and it was depicted that the concordance rate between CISH and qPCR assays was 88.1%.The quantification of HER2 gene with CISH and qPCR showed that there was a significant correlation (p value= 0.0001 and r= 0.808). The results of this research support the idea that qPCR is a precise and reproducible technique, which can be employed as a supplementary method to evaluate HER2 status.

  3. Medium modification of fragmentation functions

    International Nuclear Information System (INIS)

    Nezza, Pasquale Di

    2007-01-01

    Deep Inelastic Scattering is the cleanest process to investigate the space-time evolution of the hadronization. This was studied by the influence of the nuclear medium on lepto-production of hadrons at the Hermes experiment at DESY in semi-inclusive DIS of 27.6 GeV positrons off deuterium, nitrogen, krypton and xenon targets. The differential multiplicity for heavy targets relative to that of deuterium has been measured for the first time for various identified hadrons (φ + , φ - , φ 0 , k + , k - , p and anti-p) as a function of the virtual photon energy ?, the fraction z of this energy transferred to the hadron, and the hadron transverse momentum squared p 2 t . The distribution of the hadron transverse momentum is broadened towards high p 2 t in the nuclear medium, in a manner resembling the Cronin effect observed in collisions of heavy ions and protons with nuclei. The pt -broadening results give also important information about the pre-hadron formation time. Moreover, by studying the hadron attenuation of the leading and sub-leading hadrons, we report, for the first time, the possibility to better understand the hadron absorption and the energy loss contributions to the attenuation mechanism. (Author)

  4. Primordial condensation of meteorite components - experimental evidence of the state of the source medium

    International Nuclear Information System (INIS)

    Arrhenius, G.; McCrumb, J.L.; Friedman, N.

    1979-01-01

    Mineral grains and grain aggregates in meteorites carry potential information on the conditions in the environment where they formed. To avoid model-dependent interpretations it is necessary to develop experimental criteria that uniquely reflect the environmental parameters of interest. These parameters include the various temperatures of the source medium and the temperature of grains at growth all of which are observed to be highly differentiated in the space medium in accordance with the radiation laws. (orig./WL)

  5. Merger of waste in kaolin panels medium density

    International Nuclear Information System (INIS)

    Bezerra, A.F.C.; Santana, L.N.L.; Neves, G.A.

    2011-01-01

    Medium-density panels are molded under pressure and temperature and have physical and mechanical properties similar to those of solid wood. Their composition involves fibers of eucalyptus and pine, but other residues as kaolin waste can be incorporated. The objective was to manufacture medium density panels incorporating kaolin waste and compare the physical, chemical and mechanical properties of these with other commercials. The residue was subjected to the following characterization tests: X-ray diffraction, chemical analysis, differential thermal analysis, thermal gravimetric analysis and size analysis.Through the process of pressing the samples were prepared, they were evaluated for their flexural strength and tensile strength perpendicular to the water absorption / swelling in thickness, density and moisture content. According to the analyzed results, we conclude that samples having the residue had lower levels of swelling, tensile and flexural strength and higher levels of absorption.(author)

  6. Fem Formulation for Heat and Mass Transfer in Porous Medium

    Science.gov (United States)

    Azeem; Soudagar, Manzoor Elahi M.; Salman Ahmed, N. J.; Anjum Badruddin, Irfan

    2017-08-01

    Heat and mass transfer in porous medium can be modelled using three partial differential equations namely, momentum equation, energy equation and mass diffusion. These three equations are coupled to each other by some common terms that turn the whole phenomenon into a complex problem with inter-dependable variables. The current article describes the finite element formulation of heat and mass transfer in porous medium with respect to Cartesian coordinates. The problem under study is formulated into algebraic form of equations by using Galerkin's method with the help of two-node linear triangular element having three nodes. The domain is meshed with smaller sized elements near the wall region and bigger size away from walls.

  7. In-medium ϕ meson width extracted from proton-nucleus collisions

    Directory of Open Access Journals (Sweden)

    Roca L.

    2012-12-01

    Full Text Available The inclusive production of ϕ mesons at small angles in proton collisions with C, Cu, Ag, and Au targets has been measured at an incident energy of 2.83 GeV at the ANKE-COSY facility. The ϕ mesons were registered via the ϕ → K+K− decay. The momentum dependence of the nuclear transparency ratio, the in-medium ϕ width, and the differential cross section for ϕ production have been determined in the momentum region pϕ = 0.6 − 1.6 GeV/c. Comparison with different model calculations suggests a significant broadening of the in-medium ϕ width for normal nuclear density with evidence for a momentum dependence.

  8. [Molecular classification of breast cancer patients obtained through the technique of chromogenic in situ hybridization (CISH)].

    Science.gov (United States)

    Fernández, Angel; Reigosa, Aldo

    2013-12-01

    Breast cancer is a heterogeneous disease composed of a growing number of biological subtypes, with substantial variability of the disease progression within each category. The aim of this research was to classify the samples object of study according to the molecular classes of breast cancer: luminal A, luminal B, HER2 and triple negative, as a result of the state of HER2 amplification obtained by the technique of chromogenic in situ hybridization (CISH). The sample consisted of 200 biopsies fixed in 10% formalin, processed by standard techniques up to paraffin embedding, corresponding to patients diagnosed with invasive ductal carcinoma of the breast. These biopsies were obtained from patients from private practice and the Institute of Oncology "Dr. Miguel Pérez Carreño", for immunohistochemistry (IHC) of hormone receptors and HER2 made in the Hospital Metropolitano del Norte, Valencia, Venezuela. The molecular classification of the patient's tumors considering the expression of estrogen and progesterone receptors by IHC and HER2 amplification by CISH, allowed those cases originally classified as unknown, since they had an indeterminate (2+) outcome for HER2 expression by IHC, to be grouped into the different molecular classes. Also, this classification permitted that some cases, initially considered as belonging to a molecular class, were assigned to another class, after the revaluation of the HER2 status by CISH.

  9. Dielectric metasurfaces solve differential and integro-differential equations.

    Science.gov (United States)

    Abdollahramezani, Sajjad; Chizari, Ata; Dorche, Ali Eshaghian; Jamali, Mohammad Vahid; Salehi, Jawad A

    2017-04-01

    Leveraging subwavelength resonant nanostructures, plasmonic metasurfaces have recently attracted much attention as a breakthrough concept for engineering optical waves both spatially and spectrally. However, inherent ohmic losses concomitant with low coupling efficiencies pose fundamental impediments over their practical applications. Not only can all-dielectric metasurfaces tackle such substantial drawbacks, but also their CMOS-compatible configurations support both Mie resonances that are invariant to the incident angle. Here, we report on a transmittive metasurface comprising arrayed silicon nanodisks embedded in a homogeneous dielectric medium to manipulate phase and amplitude of incident light locally and almost independently. By taking advantage of the interplay between the electric/magnetic resonances and employing general concepts of spatial Fourier transformation, a highly efficient metadevice is proposed to perform mathematical operations including solution of ordinary differential and integro-differential equations with constant coefficients. Our findings further substantiate dielectric metasurfaces as promising candidates for miniaturized, two-dimensional, and planar optical analog computing systems that are much thinner than their conventional lens-based counterparts.

  10. Propagation of mechanical waves through a stochastic medium with spherical symmetry

    Science.gov (United States)

    Avendaño, Carlos G.; Reyes, J. Adrián

    2018-01-01

    We theoretically analyze the propagation of outgoing mechanical waves through an infinite isotropic elastic medium possessing spherical symmetry whose Lamé coefficients and density are spatial random functions characterized by well-defined statistical parameters. We derive the differential equation that governs the average displacement for a system whose properties depend on the radial coordinate. We show that such an equation is an extended version of the well-known Bessel differential equation whose perturbative additional terms contain coefficients that depend directly on the squared noise intensities and the autocorrelation lengths in an exponential decay fashion. We numerically solve the second order differential equation for several values of noise intensities and autocorrelation lengths and compare the corresponding displacement profiles with that of the exact analytic solution for the case of absent inhomogeneities.

  11. Genetic exchange versus genetic differentiation in a medium-sized inversion of Drosophila: the A2/Ast arrangements of Drosophila subobscura.

    Science.gov (United States)

    Nóbrega, Clévio; Khadem, Mahnaz; Aguadé, Montserrat; Segarra, Carmen

    2008-08-01

    Chromosomal inversion polymorphism affects nucleotide variation at loci associated with inversions. In Drosophila subobscura, a species with a rich chromosomal inversion polymorphism and the largest recombinational map so far reported in the Drosophila genus, extensive genetic structure of nucleotide variation was detected in the segment affected by the O(3) inversion, a moderately sized inversion at Muller's element E. Indeed, a strong genetic differentiation all over O(3) and no evidence of a higher genetic exchange in the center of the inversion than at breakpoints were detected. In order to ascertain, whether other polymorphic and differently sized inversions of D. subobscura also exhibited a strong genetic structure, nucleotide variation in 5 gene regions (P236, P275, P150, Sxl, and P125) located along the A(2) inversion was analyzed in A(st) and A(2) chromosomes of D. subobscura. A(2) is a medium-sized inversion at Muller's element A and forms a single inversion loop in heterokaryotypes. The lower level of variation in A(2) relative to A(st) and the significant excess of low-frequency variants at polymorphic sites indicate that nucleotide variation at A(2) is not at mutation-drift equilibrium. The closest region to an inversion breakpoint, P236, exhibits the highest level of genetic differentiation (F(ST)) and of linkage disequilibrium (LD) between arrangements and variants at nucleotide polymorphic sites. The remaining 4 regions show a higher level of genetic exchange between A(2) and A(st) chromosomes than P236, as revealed by F(ST) and LD estimates. However, significant genetic differentiation between the A(st) and A(2) arrangements was detected not only at P236 but also in the other 4 regions separated from the nearest breakpoint by 1.2-2.9 Mb. Therefore, the extent of genetic exchange between arrangements has not been high enough to homogenize nucleotide variation in the center of the A(2) inversion. A(2) can be considered a typical successful inversion

  12. Fluorescence (FISH) and chromogenic (CISH) in situ hybridisation in prostate carcinoma cell lines: comparison and use of virtual microscopy.

    Science.gov (United States)

    Elliott, K; Hamilton, P W; Maxwell, P

    2008-01-01

    Chromogenic in situ hybridisation (CISH) has become an attractive alternative to fluorescence in situ hybridisation (FISH) due to its permanent stain which is more familiar to pathologists and because it can be viewed using light microscopy. The aim of the present study is to examine reproducibility in the assessment of abnormal chromosome number by CISH in comparison to FISH. Using three prostate cell lines--PNT1A (derived from normal epithelium), LNCAP and DU145 (derived from prostatic carcinoma), chromosomes 7 and 8 were counted in 40 nuclei in FISH preparations (x100 oil immersion) and 100 nuclei in CISH preparations (x40) by two independent observers. The CISH slides were examined using standard light microscopy and virtual microscopy. Reproducibility was examined using paired Student's t-test (PCISH. No significant differences in chromosome count were seen between the techniques. Chromosomes 7 and 8 showed disomic status for each cell line except LNCAP, which proved to be heterogeneous (disomic/aneusomic), particularly for chromosome 8. Virtual microscopy proved to be easy to use and gave no significant differences from standard light microscopy. These results support the hypothesis that there is no significant difference between FISH and CISH techniques.

  13. Determination of HER2 amplification in primary breast cancer using dual-colour chromogenic in situ hybridization is comparable to fluorescence in situ hybridization: a European multicentre study involving 168 specimens

    Science.gov (United States)

    García-Caballero, Tomás; Grabau, Dorthe; Green, Andrew R; Gregory, John; Schad, Arno; Kohlwes, Elke; Ellis, Ian O; Watts, Sarah; Mollerup, Jens

    2010-01-01

    García-Caballero T, Grabau D, Green A R, Gregory J, Schad A, Kohlwes E, Ellis I O, Watts S & Mollerup J (2010) Histopathology56, 472–480 Determination of HER2 amplification in primary breast cancer using dual-colour chromogenic in situ hybridization is comparable to fluorescence in situ hybridization: a European multicentre study involving 168 specimens Aims: Fluorescence in situ hybridization (FISH) can be used to reveal several genomic imbalances relevant to proper cancer diagnosis and to the correct treatment regime. However, FISH requires expensive and advanced fluorescence microscopes in addition to expertise in fluorescence microscopy. To determine whether a newly developed dual-colour chromogenic in situ hybridization (CISH) method is a suitable alternative to FISH, we analysed the human epidermal growth factor receptor 2 gene (HER2) amplification level of 168 breast cancer specimens using dual-colour CISH and FISH and compared the results. Methods and results: We found 100% agreement between HER2 status determined by FISH and dual-colour CISH. Furthermore, we observed that the time used to score slides was significantly reduced by 28% in dual-colour CISH compared with the FISH protocol. Concordance between HER2 protein status and dual-colour CISH or FISH was equally good with an overall agreement of 96.8%. Correlation between the HER2/centromere 17 gene ratios obtained with dual-colour CISH and FISH was highly significant with an overall correlation coefficient (ρ) of 0.96. Conclusions: We conclude that dual-colour CISH and bright field microscopy are excellent alternatives to FISH when analysing the HER2 status of primary breast cancer. PMID:20459554

  14. Instantaneous Real-Time Kinematic Decimeter-Level Positioning with BeiDou Triple-Frequency Signals over Medium Baselines.

    Science.gov (United States)

    He, Xiyang; Zhang, Xiaohong; Tang, Long; Liu, Wanke

    2015-12-22

    Many applications, such as marine navigation, land vehicles location, etc., require real time precise positioning under medium or long baseline conditions. In this contribution, we develop a model of real-time kinematic decimeter-level positioning with BeiDou Navigation Satellite System (BDS) triple-frequency signals over medium distances. The ambiguities of two extra-wide-lane (EWL) combinations are fixed first, and then a wide lane (WL) combination is reformed based on the two EWL combinations for positioning. Theoretical analysis and empirical analysis is given of the ambiguity fixing rate and the positioning accuracy of the presented method. The results indicate that the ambiguity fixing rate can be up to more than 98% when using BDS medium baseline observations, which is much higher than that of dual-frequency Hatch-Melbourne-Wübbena (HMW) method. As for positioning accuracy, decimeter level accuracy can be achieved with this method, which is comparable to that of carrier-smoothed code differential positioning method. Signal interruption simulation experiment indicates that the proposed method can realize fast high-precision positioning whereas the carrier-smoothed code differential positioning method needs several hundreds of seconds for obtaining high precision results. We can conclude that a relatively high accuracy and high fixing rate can be achieved for triple-frequency WL method with single-epoch observations, displaying significant advantage comparing to traditional carrier-smoothed code differential positioning method.

  15. Instantaneous Real-Time Kinematic Decimeter-Level Positioning with BeiDou Triple-Frequency Signals over Medium Baselines

    Directory of Open Access Journals (Sweden)

    Xiyang He

    2015-12-01

    Full Text Available Many applications, such as marine navigation, land vehicles location, etc., require real time precise positioning under medium or long baseline conditions. In this contribution, we develop a model of real-time kinematic decimeter-level positioning with BeiDou Navigation Satellite System (BDS triple-frequency signals over medium distances. The ambiguities of two extra-wide-lane (EWL combinations are fixed first, and then a wide lane (WL combination is reformed based on the two EWL combinations for positioning. Theoretical analysis and empirical analysis is given of the ambiguity fixing rate and the positioning accuracy of the presented method. The results indicate that the ambiguity fixing rate can be up to more than 98% when using BDS medium baseline observations, which is much higher than that of dual-frequency Hatch-Melbourne-Wübbena (HMW method. As for positioning accuracy, decimeter level accuracy can be achieved with this method, which is comparable to that of carrier-smoothed code differential positioning method. Signal interruption simulation experiment indicates that the proposed method can realize fast high-precision positioning whereas the carrier-smoothed code differential positioning method needs several hundreds of seconds for obtaining high precision results. We can conclude that a relatively high accuracy and high fixing rate can be achieved for triple-frequency WL method with single-epoch observations, displaying significant advantage comparing to traditional carrier-smoothed code differential positioning method.

  16. Characteristic bacteriolytic activities of Staphylococcus hyicus.

    OpenAIRE

    Lämmler, C

    1989-01-01

    Staphylococcus hyicus demonstrated characteristic bacteriolytic activities towards a Micrococcus luteus reference strain. This lytic activity was demonstrated on medium containing M. luteus cells as large zones of transparency around the culture streak. Smaller zones of transparency were observed with Staphylococcus intermedius, Staphylococcus chromogenes, and some strains of Staphylococcus aureus but not with other coagulase-negative staphylococcal species. The distribution and extent of the...

  17. Simple, Reliable, and Cost-Effective Yeast Identification Scheme for the Clinical Laboratory

    OpenAIRE

    Koehler, Ann P.; Chu, Kai-Cheong; Houang, Elizabeth T. S.; Cheng, Augustine F. B.

    1999-01-01

    The appearance of colonies on the chromogenic medium CHROMagar Candida combined with observation of morphology on corn meal–Tween 80 agar was used for the identification of 353 clinical yeast isolates. The results were compared with those obtained with API yeast identification kits. The accuracy of identification and the turnaround time were equivalent for each method, and our cultural method was less expensive.

  18. A Newly Defined and Xeno-Free Culture Medium Supports Every-Other-Day Medium Replacement in the Generation and Long-Term Cultivation of Human Pluripotent Stem Cells.

    Science.gov (United States)

    Ahmadian Baghbaderani, Behnam; Tian, Xinghui; Scotty Cadet, Jean; Shah, Kevan; Walde, Amy; Tran, Huan; Kovarcik, Don Paul; Clarke, Diana; Fellner, Thomas

    2016-01-01

    Human pluripotent stem cells (hPSCs) present an unprecedented opportunity to advance human health by offering an alternative and renewable cell resource for cellular therapeutics and regenerative medicine. The present demand for high quality hPSCs for use in both research and clinical studies underscores the need to develop technologies that will simplify the cultivation process and control variability. Here we describe the development of a robust, defined and xeno-free hPSC medium that supports reliable propagation of hPSCs and generation of human induced pluripotent stem cells (hiPSCs) from multiple somatic cell types; long-term serial subculturing of hPSCs with every-other-day (EOD) medium replacement; and banking fully characterized hPSCs. The hPSCs cultured in this medium for over 40 passages are genetically stable, retain high expression levels of the pluripotency markers TRA-1-60, TRA-1-81, Oct-3/4 and SSEA-4, and readily differentiate into ectoderm, mesoderm and endoderm. Importantly, the medium plays an integral role in establishing a cGMP-compliant process for the manufacturing of hiPSCs that can be used for generation of clinically relevant cell types for cell replacement therapy applications.

  19. Prognostic value of HER2 gene amplification detected by chromogenic in situ hybridization (CISH) in metastatic breast cancer.

    Science.gov (United States)

    Todorović-Raković, Natasa; Jovanović, Danica; Nesković-Konstantinović, Zora; Nikolić-Vukosavljević, Dragica

    2007-06-01

    After so many years of research, clinical value of HER2 (Human epidermal growth factor receptor 2) is unclear. Perhaps the main reason is variability of testing methods that produce controversial results. There is a lack of studies regarding prognostic value of CISH especially in metastatic breast cancer (MBC) when risk evaluation is based on different parameters than for primary breast cancer. Aim of this study was to compare prognostic relevance of HER2 status in MBC tested by two different methods i.e. immunohistochemistry (IHC) and chromogenic in situ hybridization (CISH). HER2 status of the same group of 107 MBC patients was determined by IHC (protein overexpression) and by CISH (gene amplification). HER2 results obtained by IHC and CISH showed significant correlation, beside the existence of discrepancies. Beside the significant correlation in two methods, there was a difference in prognostic values of compared methods during the course of metastatic disease. There was a significant difference in progression-free interval (PFI) between HER2 non-amplified and HER2 amplified cases determined by CISH, in postmenopausal subgroup and node-positive subgroup, but no significant difference for IHC stratified MBC patients. CISH seems to be accurate and more informative method than IHC regarding prognostic value of HER2 in metastatic breast cancer.

  20. Calculation of double energy angle differential neutron albedos for radiation shielding applications

    International Nuclear Information System (INIS)

    Litaize, O.; Diop, C.M.; Nimal, J.C.

    2000-01-01

    Void radiation shielding problems can be dealt with albedo concept which is an alternative to the complex bringing into operation of the 'exact' transport method calculations (SN, Monte Carlo). Up to here, differential albedos are used for single reflections from walls in the NARCISSE-3 propagation albedo code developed at CEA and used for project calculations. For taking into account the neutron multiple reflections on lacunar medium walls, double energy-angle differential albedos are needed. TRIPOLI-4 neutral particle transport Monte Carlo code in three dimensional geometries, has been chosen to implement a double differential albedo calculus routine and therefore to generate albedo data for different kinds of medium. The surfacic estimator, which could be used, is not enough efficient because all neutrons do not contribute to the result. A new estimator is carried out. At each collision site, during the neutron history simulation, it allows to compute the probability of the neutron to go through the medium and to come through the reflection surface in the direction and at the energy considered. This estimator is about hundred times more efficient than the surfacic estimator. (author)

  1. Characterization of a Novel Subgroup of Extracellular Medium-Chain-Length Polyhydroxyalkanoate Depolymerases from Actinobacteria

    Science.gov (United States)

    Gangoiti, Joana; Santos, Marta; Prieto, María Auxiliadora; de la Mata, Isabel; Llama, María J.

    2012-01-01

    Nineteen medium-chain-length (mcl) poly(3-hydroxyalkanoate) (PHA)-degrading microorganisms were isolated from natural sources. From them, seven Gram-positive and three Gram-negative bacteria were identified. The ability of these microorganisms to hydrolyze other biodegradable plastics, such as short-chain-length (scl) PHA, poly(ε-caprolactone) (PCL), poly(ethylene succinate) (PES), and poly(l-lactide) (PLA), has been studied. On the basis of the great ability to degrade different polyesters, Streptomyces roseolus SL3 was selected, and its extracellular depolymerase was biochemically characterized. The enzyme consisted of one polypeptide chain of 28 kDa with a pI value of 5.2. Its maximum activity was observed at pH 9.5 with chromogenic substrates. The purified enzyme hydrolyzed mcl PHA and PCL but not scl PHA, PES, and PLA. Moreover, the mcl PHA depolymerase can hydrolyze various substrates for esterases, such as tributyrin and p-nitrophenyl (pNP)-alkanoates, with its maximum activity being measured with pNP-octanoate. Interestingly, when poly(3-hydroxyoctanoate-co-3-hydroxyhexanoate [11%]) was used as the substrate, the main hydrolysis product was the monomer (R)-3-hydroxyoctanoate. In addition, the genes of several Actinobacteria strains, including S. roseolus SL3, were identified on the basis of the peptide de novo sequencing of the Streptomyces venezuelae SO1 mcl PHA depolymerase by tandem mass spectrometry. These enzymes did not show significant similarity to mcl PHA depolymerases characterized previously. Our results suggest that these distinct enzymes might represent a new subgroup of mcl PHA depolymerases. PMID:22865072

  2. Detection of Epstein Barr Virus by Chromogenic In Situ Hybridization in cases of extra-hepatic biliary atresia

    Directory of Open Access Journals (Sweden)

    Farahmand Fatemeh

    2008-04-01

    Full Text Available Abstract Introduction Extra-hepatic biliary atresia (EHBA is an important cause of neonatal cholestasis. Several infectious agents have been proposed as etiologic factors such as Rotavirus and Reovirus. There is limited data on the role of Epstein Barr virus (EBV infection in EHBA, so we decided to study the presence of EBV virus in a series of 16 proven EHBA cases by Chromogenic in situ hybridization (CISH technique. Methods In the current study a total of 16 liver wedge biopsies of proven cases of EHBA were selected in a period of 4 years. CISH staining for EBV-encoded RNA (EBER transcript was performed. Results The review of H&E-stained slides of liver biopsies revealed fibrosis and marked ductular proliferation. In CISH-stained slides, EBV trace was observed in hepatocytes in two cases and in biliary epithelium in one case of EHBA. Discussion Considering the association of hepatitis with the Epstein-Barr virus in later life, it is likely that EBV hepatitis and its complications occur in the neonatal/perinatal period. Since EHBA is a relatively rare disease, a similar study on wedge biopsies of this number of proven cases of EHBA has not been performed to date. Current observation proposes the need for a study of larger series and employing other methods for confirming the etiologic role of EBV in EHBA cases.

  3. Heat transfer in porous medium embedded with vertical plate: Non-equilibrium approach - Part A

    Energy Technology Data Exchange (ETDEWEB)

    Badruddin, Irfan Anjum [Dept. of Mechanical Engineering, University of Malaya, Kuala Lumpur, 50603 (Malaysia); Quadir, G. A. [School of Mechatronic Engineering, University Malaysia Perlis, Pauh Putra, 02600 Arau, Perlis (Malaysia)

    2016-06-08

    Heat transfer in a porous medium embedded with vertical flat plate is investigated by using thermal non-equilibrium model. Darcy model is employed to simulate the flow inside porous medium. It is assumed that the heat transfer takes place by natural convection and radiation. The vertical plate is maintained at isothermal temperature. The governing partial differential equations are converted into non-dimensional form and solved numerically using finite element method. Results are presented in terms of isotherms and streamlines for various parameters such as heat transfer coefficient parameter, thermal conductivity ratio, and radiation parameter.

  4. Heat transfer in porous medium embedded with vertical plate: Non-equilibrium approach - Part A

    International Nuclear Information System (INIS)

    Badruddin, Irfan Anjum; Quadir, G. A.

    2016-01-01

    Heat transfer in a porous medium embedded with vertical flat plate is investigated by using thermal non-equilibrium model. Darcy model is employed to simulate the flow inside porous medium. It is assumed that the heat transfer takes place by natural convection and radiation. The vertical plate is maintained at isothermal temperature. The governing partial differential equations are converted into non-dimensional form and solved numerically using finite element method. Results are presented in terms of isotherms and streamlines for various parameters such as heat transfer coefficient parameter, thermal conductivity ratio, and radiation parameter

  5. Differentiation of the entomopathogenic fungus Metarhizium flavoviride (Hyphomycetes

    Directory of Open Access Journals (Sweden)

    Xavier-Santos Solange

    1999-01-01

    Full Text Available The differentiation of a Brazilian isolate of Metarhizium flavoviride (CG 423, a promising candidate for the biocontrol of grasshoppers, was investigated. Conidia were spread onto solid medium (1% yeast extract, 2.8% agar, 96.2% distilled water, incubated at 28°C and observed during 26 h. Germination initiated as conidia size increased from 5.3 (±0.6 x 3.1 (±0.3 µm (0 h incubation to 8.1 (±0.2 x 6.1 (±0.2 µm (8 h incubation. Germ tubes started to appear after 10 h incubation showing a high degree of multipolarity. Twenty six hours after inoculation, hyphal differentiation and anastomosis among hyphae from adjacent conidia were recorded. Appressoria were formed only from conidia incubated in liquid medium containing minimum concentration of yeast extract (0.06%; w/v. Appressoria were firmly adhered to the bottom of plastic dishes.

  6. Micro-Topographies Promote Late Chondrogenic Differentiation Markers in the ATDC5 Cell Line.

    Science.gov (United States)

    Le, Bach Q; Vasilevich, Aliaksei; Vermeulen, Steven; Hulshof, Frits; Stamatialis, Dimitrios F; van Blitterswijk, Clemens A; de Boer, Jan

    2017-05-01

    Chemical and mechanical cues are well-established influencers of in vitro chondrogenic differentiation of ATDC5 cells. Here, we investigate the role of topographical cues in this differentiation process, a study not been explored before. Previously, using a library of surface micro-topographies we found some distinct patterns that induced alkaline phosphatase (ALP) production in human mesenchymal stromal cells. ALP is also a marker for hypertrophy, the end stage of chondrogenic differentiation preceding bone formation. Thus, we hypothesized that these patterns could influence end-stage chondrogenic differentiation of ATDC5 cells. In this study, we randomly selected seven topographies among the ALP influencing hits. Cells grown on these surfaces displayed varying nuclear shape and actin filament structure. When stimulated with insulin-transferrin-selenium (ITS) medium, nodule formation occurred and in some cases showed alignment to the topographical patterns. Gene expression analysis of cells growing on topographical surfaces in the presence of ITS medium revealed a downregulation of early markers and upregulation of late markers of chondrogenic differentiation compared to cells grown on a flat surface. In conclusion, we demonstrated that surface topography in addition to other cues can promote hypertrophic differentiation suitable for bone tissue engineering.

  7. Transient response of a cylindrical cavity in viscoelastic saturated porous medium

    Directory of Open Access Journals (Sweden)

    LIU Tao

    2016-10-01

    Full Text Available The study on dynamic characteristics for fluid-solid coupling system in saturated porous medium is of significant academic value and potential application foreground.In this paper,the transient response of a cylindrical cavity in infinite viscoelastic saturated porous medium with the circular lining is studied,and the corresponding results can be used in the design of foundation engineering,such as the tunnel analyses in saturated soil,the nuclear waste disposal engineering,and the exploitation and utilization of geothermal reservoirs and so on.Firstly,based on the porous media theory,the governing equations of coupled system are presented,and the corresponding boundary conditions,initial conditions as well as the joint conditions are derived.Then,the differential quadrature element method and the second-order backward difference scheme are applied to discretize the governing differential equations of the coupled system on the spatial and temporal domains,respectively.Finally,the Newton-Raphson method is adopted to solve the discretization equations with the initial conditions,the transient responses of the coupled system are analyzed,the effects of the parameters are considered,and the validity of the numerical method is verified.

  8. Bleaching of melanin in the epidermis of South American fur seal and its application on enzyme immunohistochemistry

    Directory of Open Access Journals (Sweden)

    A.P. Silva

    2011-03-01

    Full Text Available The South American fur seal (Arctocephalus australis is an amphibious marine mammal distributed along the Atlantic and Pacific coasts of South America. The species is well adjusted to different habitats due to the morphology of its fin-like members and due to some adaptations in their integumentary system. Immunohistochemical studies are very important to evaluate the mechanisms of skin adaptation due the differential expression of the antigens present in the tissue depending of the region of the body surface. However, its strongly pigmented (melanin epidermis prevents the visualization of the immuno-histochemical chromogens markers. In this study a melanin bleaching method was developed aimed to allow the visualization of the chromogens without interfering in the antigen-antibody affinity for immunohistochemistry. The analysis of PCNA (proliferating cell nuclear antigen index in the epidermis of A. australis by immunohistochemistry with diaminobenzidine (DAB as chromogen was used to test the method. The bleaching of the melanin allowed to obtain the cell proliferation index in epidermis and to avoid false positive results without affecting the immunohistochemical results.

  9. ACOUSTIC WAVES EMISSION IN THE TWO-COMPONENT HEREDITARY-ELASTIC MEDIUM

    Directory of Open Access Journals (Sweden)

    V. S. Polenov

    2014-01-01

    Full Text Available Summary. On the dynamics of two-component media a number of papers, which address the elastic waves in a homogeneous, unbounded fluid-saturated porous medium. In other studies address issues of dissipative processes in harmonic deformation hereditary elastic medium. In the article the dissipative processes of the viscoelastic porous medium, which hereditary properties are described by the core relaxation fractional exponential function U.N. Rabotnova integro-differential Boltzmann-Volterr ratio, harmonic deformation by the straining saturated incompressible liquid are investigated. Speed of wave propagation, absorption coefficient, mechanical loss tangent, logarithmic decrement, depending on fractional parameter γ, determining formulas received. The frequency logarithm and temperature graph dependences with the goal fractional parameter are constructed. Shows the dependences velocity and attenuation coefficient of the tangent of the phase angle of the logarithm of the temperature, and the dependence of the attenuation coefficient of the logarithm of the frequency. Dependencies the speed and the tangent of the phase angle of the frequency identical function of the logarithm of temperature.

  10. Changes in the chromogen properties of the betalaine induced by gamma radiation; Cambios en las propiedades cromogenas de las betalainas inducidos por radiacion gamma

    Energy Technology Data Exchange (ETDEWEB)

    Rodriguez N, S.; Pinedo S, A.; Amador V, P.; Chacon R, A.; Arcos P, A.; Vega C, H.R. [Unidad Academica de Ciencias Nucleares, C. Cipres 10, Fracc. La Penuela, 98068 Zacatecas (Mexico)

    2005-07-01

    The changes of coloration of four natural extracts in function of the absorbed dose produced by a gamma rays source of {sup 137} Cs were determined. The natural extracts used contain betalaines that are natural pigments of some plants as the beet about that their name. They are also in abundant form in the fruits (tunas) of some species of the Opuntia genus. The extracts were obtained by maceration, starting from beet and three tuna varieties, and they were stabilized to pH 5.5 The change of coloration it was determined in a visible ultraviolet spectrophotometer by means of the absorbance from the samples to photons of 475, 535 and 600 nm of wavelength. The absorbance to different intervals of time was measured. The relationship between the absorbed dose and the chromogene properties of the pigment, with the intention of using it as possible dosemeter was settled down. (Author)

  11. Differentiation of isolated human umbilical cord mesenchymal stem cells into neural stem cells

    Science.gov (United States)

    Chen, Song; Zhang, Wei; Wang, Ji-Ming; Duan, Hong-Tao; Kong, Jia-Hui; Wang, Yue-Xin; Dong, Meng; Bi, Xue; Song, Jian

    2016-01-01

    AIM To investigate whether umbilical cord human mesenchymal stem cell (UC-MSC) was able to differentiate into neural stem cell and neuron in vitro. METHODS The umbilical cords were obtained from pregnant women with their written consent and the approval of the Clinic Ethnics Committee. UC-MSC were isolated by adherent culture in the medium contains 20% fetal bovine serum (FBS), then they were maintained in the medium contain 10% FBS and induced to neural cells in neural differentiation medium. We investigated whether UC-MSC was able to differentiate into neural stem cell and neuron in vitro by using flow cytometry, reverse transcriptase-polymerase chain reaction (RT-PCR) and immunofluorescence (IF) analyzes. RESULTS A substantial number of UC-MSC was harvested using the tissue explants adherent method at about 2wk. Flow cytometric study revealed that these cells expressed common markers of MSCs, such as CD105 (SH2), CD73 (SH3) and CD90. After induction of differentiation of neural stem cells, the cells began to form clusters; RT-PCR and IF showed that the neuron specific enolase (NSE) and neurogenic differentiation 1-positive cells reached 87.3%±14.7% and 72.6%±11.8%, respectively. Cells showed neuronal cell differentiation after induced, including neuron-like protrusions, plump cell body, obviously and stronger refraction. RT-PCR and IF analysis showed that microtubule-associated protein 2 (MAP2) and nuclear factor-M-positive cells reached 43.1%±10.3% and 69.4%±19.5%, respectively. CONCLUSION Human umbilical cord derived MSCs can be cultured and proliferated in vitro and differentiate into neural stem cells, which may be a valuable source for cell therapy of neurodegenerative eye diseases. PMID:26949608

  12. Differentiation of isolated human umbilical cord mesenchymal stem cells into neural stem cells

    Directory of Open Access Journals (Sweden)

    Song Chen

    2016-01-01

    Full Text Available AIM: To investigate whether umbilical cord human mesenchymal stem cell (UC-MSC was able to differentiate into neural stem cell and neuron in vitro. METHODS: The umbilical cords were obtained from pregnant women with their written consent and the approval of the Clinic Ethnics Committee. UC-MSC were isolated by adherent culture in the medium contains 20% fetal bovine serum (FBS, then they were maintained in the medium contain 10% FBS and induced to neural cells in neural differentiation medium. We investigated whether UC-MSC was able to differentiate into neural stem cell and neuron in vitro by using flow cytometry, reverse transcriptase-polymerase chain reaction (RT-PCR and immunofluorescence (IF analyzes. RESULTS: A substantial number of UC-MSC was harvested using the tissue explants adherent method at about 2wk. Flow cytometric study revealed that these cells expressed common markers of MSCs, such as CD105 (SH2, CD73 (SH3 and CD90. After induction of differentiation of neural stem cells, the cells began to form clusters; RT-PCR and IF showed that the neuron specific enolase (NSE and neurogenic differentiation 1-positive cells reached 87.3%±14.7% and 72.6%±11.8%, respectively. Cells showed neuronal cell differentiation after induced, including neuron-like protrusions, plump cell body, obviously and stronger refraction. RT-PCR and IF analysis showed that microtubule-associated protein 2 (MAP2 and nuclear factor-M-positive cells reached 43.1%±10.3% and 69.4%±19.5%, respectively. CONCLUSION: Human umbilical cord derived MSCs can be cultured and proliferated in vitro and differentiate into neural stem cells, which may be a valuable source for cell therapy of neurodegenerative eye diseases.

  13. Operator overloading as an enabling technology for automatic differentiation

    International Nuclear Information System (INIS)

    Corliss, G.F.; Griewank, A.

    1993-01-01

    We present an example of the science that is enabled by object-oriented programming techniques. Scientific computation often needs derivatives for solving nonlinear systems such as those arising in many PDE algorithms, optimization, parameter identification, stiff ordinary differential equations, or sensitivity analysis. Automatic differentiation computes derivatives accurately and efficiently by applying the chain rule to each arithmetic operation or elementary function. Operator overloading enables the techniques of either the forward or the reverse mode of automatic differentiation to be applied to real-world scientific problems. We illustrate automatic differentiation with an example drawn from a model of unsaturated flow in a porous medium. The problem arises from planning for the long-term storage of radioactive waste

  14. Differentiating between heat pain intensities: the combined effect of multiple autonomic parameters.

    Science.gov (United States)

    Treister, Roi; Kliger, Mark; Zuckerman, Galit; Goor Aryeh, Itay; Eisenberg, Elon

    2012-09-01

    Although it is well known that pain induces changes in autonomic parameters, the extent to which these changes correlate with the experience of pain is under debate. The aim of the present study was to compare a combination of multiple autonomic parameters and each parameter alone in their ability to differentiate among 4 categories of pain intensity. Tonic heat stimuli (1minute) were individually adjusted to induce no pain, low, medium, and high pain in 45 healthy volunteers. Electrocardiogram, photoplethysmogram, and galvanic skin response were recorded, and the following parameters were calculated: heart rate; heart rate variability-high frequency (0.15 to 0.4Hz) spectral power; skin conductance level; number of skin conduction fluctuations; and photoplethysmographic pulse wave amplitude. A combination of parameters was created by fitting an ordinal cumulative logit model to the data and using linear coefficients of the model. Friedman test with post-hoc Wilcoxon test were used to compare between pain intensity categories for every parameter alone and for their linear combination. All of the parameters successfully differentiated between no pain and all other pain categories. However, none of the parameters differentiated between all 3 pain categories (i.e., low and medium; medium and high; low and high). In contrast, the linear combination of parameters significantly differentiated not only between pain and no pain, but also between all pain categories (Ppain assessment. Copyright © 2012 International Association for the Study of Pain. Published by Elsevier B.V. All rights reserved.

  15. Contribution to differential diagnosis of lumbar spine disc hernia by computerized tomography

    International Nuclear Information System (INIS)

    Dolansky, J.

    1989-01-01

    The significance is discussed of computerized tomography in differential diagnosis of disc hernias of the lumbar spine. A simple technique is described that allows to differentiate disc hernias from epidural vein varices. It mostly includes repeat imaging of the same body section and/or the application of a contrast medium. (L.O.). 2 figs., 4 refs

  16. Gene expression profile in human induced pluripotent stem cells: Chondrogenic differentiation in vitro, part A

    Science.gov (United States)

    Suchorska, Wiktoria Maria; Augustyniak, Ewelina; Richter, Magdalena; Trzeciak, Tomasz

    2017-01-01

    Human induced pluripotent stem cells (hiPSCs) offer promise in regenerative medicine, however more data are required to improve understanding of key aspects of the cell differentiation process, including how specific chondrogenic processes affect the gene expression profile of chondrocyte-like cells and the relative value of cell differentiation markers. The main aims of the present study were as follows: To determine the gene expression profile of chondrogenic-like cells derived from hiPSCs cultured in mediums conditioned with HC-402-05a cells or supplemented with transforming growth factor β3 (TGF-β3), and to assess the relative utility of the most commonly used chondrogenic markers as indicators of cell differentiation. These issues are relevant with regard to the use of human fibroblasts in the reprogramming process to obtain hiPSCs. Human fibroblasts are derived from the mesoderm and thus share a wide range of properties with chondrocytes, which also originate from the mesenchyme. Thus, the exclusion of dedifferentiation instead of chondrogenic differentiation is crucial. The hiPSCs were obtained from human primary dermal fibroblasts during a reprogramming process. Two methods, both involving embryoid bodies (EB), were used to obtain chondrocytes from the hiPSCs: EBs formed in a chondrogenic medium supplemented with TGF-β3 (10 ng/ml) and EBs formed in a medium conditioned with growth factors from HC-402-05a cells. Based on immunofluorescence and reverse transcription-quantiative polymerase chain reaction analysis, the results indicated that hiPSCs have the capacity for effective chondrogenic differentiation, in particular cells differentiated in the HC-402-05a-conditioned medium, which present morphological features and markers that are characteristic of mature human chondrocytes. By contrast, cells differentiated in the presence of TGF-β3 may demonstrate hypertrophic characteristics. Several genes [paired box 9, sex determining region Y-box (SOX) 5, SOX6

  17. Osteocytes Specific GSK3 Inhibition Affects In Vitro Osteogenic Differentiation

    Directory of Open Access Journals (Sweden)

    Jessika Bertacchini

    2018-05-01

    Full Text Available Osteocytes, the most important regulators of bone processes, are producers of molecules (usually proteins that act as signals in order to communicate with nearby cells. These factors control cell division (proliferation, differentiation, and survival. Substantial evidence showed different signaling pathways activated by osteocytes and involved in osteoblast differentiation, in particular in the last decade, when the Wingless-related integration site (WNT pathway assumed a critical large importance. WNT activation by inhibiting glycogen synthase kinase 3 (GSK-3 causes bone anabolism, making GSK3 a potential therapeutic target for bone diseases. In our study, we hypothesized an important role of the osteocyte MLO-Y4 conditioned medium in controlling the differentiation process of osteoblast cell line 2T3. We found an effect of diminished differentiation capability of 2T3 upon conditioning with medium from murine long bone osteocyte-Y4 cells (MLO-Y4 pre-treated with GSK3 inhibitor CHIR2201. The novel observations of this study provide knowledge about the inhibition of GSK3 in MLO-Y4 cells. This strategy could be used as a plausible target in osteocytes in order to regulate bone resorption mediated by a loss of osteoblasts activity through a paracrine loop.

  18. Use of a primary isolation medium for recovery of methicillin-resistant Staphylococcus aureus.

    OpenAIRE

    Van Enk, R A; Thompson, K D

    1992-01-01

    Clinical specimens frequently contain methicillin-resistant Staphylococcus aureus (MRSA) isolates in low numbers or mixed with methicillin-susceptible staphylococci, which can obscure MRSA on nonselective media. By using an oxacillin-containing mannitol-salt-based selective and differential medium on 936 respiratory specimens, we recovered 45% more MRSA isolates (29 versus 20) than on nonselective media alone.

  19. Gain of chromosome 7 by chromogenic in situ hybridization (CISH) in chordomas is correlated to c-MET expression.

    Science.gov (United States)

    Walter, Beatriz A; Begnami, Maria; Valera, Vladimir A; Santi, Mariarita; Rushing, Elisabeth J; Quezado, Martha

    2011-01-01

    Chordomas are low to intermediate grade malignancies that arise from remnants of embryonic notochord. They often recur after surgery and are highly resistant to conventional adjuvant therapies. Recently, the development of effective targeted molecular therapy has been investigated in chordomas that show receptors for tyrosine kinase (RTKs) activation. Expression of specific RTKs such as Epidermal Growth Factor Receptor (EGFR) and Mesenchymal-epithelial transition factor (c-MET) in chordomas may offer valuable therapeutic options. We investigated changes in copy number of chromosome 7 and correlated it with EGFR gene status and EGFR and c-MET protein expression in 22 chordoma samples. Chromosome 7 copy number was evaluated by chromogenic in situ hybridization (CISH) and protein expression of EGFR and c-MET by immunohistochemistry. Tumors mostly showed conventional histopathologic features and were found mainly in sacral (41%) and cranial sites (54.5%). Aneusomy of chromosome 7 was seen in 73% of the samples, 62% of primary tumors and in all recurrent chordomas. EGFR and c-MET were both expressed, but only c-MET protein expression was significantly correlated with chromosome 7 aneusomy (P ≤ 0.001). c-MET overexpression may represent an early chromosome 7 alteration that could play an important role during chordoma pathogenesis. c-MET overexpression shows promise as a molecular marker of response to targeted molecular therapy in the treatment of chordomas.

  20. Medium Range Flood Forecasting for Agriculture Damage Reduction

    Science.gov (United States)

    Fakhruddin, S. H. M.

    2014-12-01

    Early warning is a key element for disaster risk reduction. In recent decades, major advancements have been made in medium range and seasonal flood forecasting. This progress provides a great opportunity to reduce agriculture damage and improve advisories for early action and planning for flood hazards. This approach can facilitate proactive rather than reactive management of the adverse consequences of floods. In the agricultural sector, for instance, farmers can take a diversity of options such as changing cropping patterns, applying fertilizer, irrigating and changing planting timing. An experimental medium range (1-10 day) flood forecasting model has been developed for Bangladesh and Thailand. It provides 51 sets of discharge ensemble forecasts of 1-10 days with significant persistence and high certainty. This type of forecast could assist farmers and other stakeholders for differential preparedness activities. These ensembles probabilistic flood forecasts have been customized based on user-needs for community-level application focused on agriculture system. The vulnerabilities of agriculture system were calculated based on exposure, sensitivity and adaptive capacity. Indicators for risk and vulnerability assessment were conducted through community consultations. The forecast lead time requirement, user-needs, impacts and management options for crops were identified through focus group discussions, informal interviews and community surveys. This paper illustrates potential applications of such ensembles for probabilistic medium range flood forecasts in a way that is not commonly practiced globally today.

  1. Early stage differentiation of thallus cells of Porphyra haitanensis (Rhodophyta)

    Science.gov (United States)

    Wang, Sujuan; Sun, Yunlong; Lu, Anming; Wang, Guangyuan

    1987-09-01

    The early stage differentiation of thallus cells of Porphyra haitanensis T. J. Chang et B. F. Zheng was studied. Protoplasts or single cells were isolated from the blades using enzyme mixture comprising 2% sea snail gut enzyme and 1% cellulase. The isolated protoplasts or single cells were incubated in the MES medium. The cell differentiations were examined under the microscope at intervals after incubation. Four types of cell differentiation, namely, normal, abnormal, carposporangial and spermatorangial, and rhizoidal types, were observed. Since normal cell differentiations occur mostly in small thalli 50 mm in length and middle portions of big thalli 200 mm in length, it is essential to select tissues from these two kinds of thalli essential for commercial production.

  2. Medium-Based Design: Extending a Medium to Create an Exploratory Learning Environment

    Science.gov (United States)

    Rick, Jochen; Lamberty, K. K.

    2005-01-01

    This article introduces "medium-based" design -- an approach to creating "exploratory learning environments" using the method of "extending a medium". First, the characteristics of exploratory learning environments and medium-based design are described and grounded in related work. Particular attention is given to "extending a medium" --…

  3. Accelerated differentiation of human induced pluripotent stem cells to blood-brain barrier endothelial cells.

    Science.gov (United States)

    Hollmann, Emma K; Bailey, Amanda K; Potharazu, Archit V; Neely, M Diana; Bowman, Aaron B; Lippmann, Ethan S

    2017-04-13

    Due to their ability to limitlessly proliferate and specialize into almost any cell type, human induced pluripotent stem cells (iPSCs) offer an unprecedented opportunity to generate human brain microvascular endothelial cells (BMECs), which compose the blood-brain barrier (BBB), for research purposes. Unfortunately, the time, expense, and expertise required to differentiate iPSCs to purified BMECs precludes their widespread use. Here, we report the use of a defined medium that accelerates the differentiation of iPSCs to BMECs while achieving comparable performance to BMECs produced by established methods. Induced pluripotent stem cells were seeded at defined densities and differentiated to BMECs using defined medium termed E6. Resultant purified BMEC phenotypes were assessed through trans-endothelial electrical resistance (TEER), fluorescein permeability, and P-glycoprotein and MRP family efflux transporter activity. Expression of endothelial markers and their signature tight junction proteins were confirmed using immunocytochemistry. The influence of co-culture with astrocytes and pericytes on purified BMECs was assessed via TEER measurements. The robustness of the differentiation method was confirmed across independent iPSC lines. The use of E6 medium, coupled with updated culture methods, reduced the differentiation time of iPSCs to BMECs from thirteen to 8 days. E6-derived BMECs expressed GLUT-1, claudin-5, occludin, PECAM-1, and VE-cadherin and consistently achieved TEER values exceeding 2500 Ω × cm 2 across multiple iPSC lines, with a maximum TEER value of 4678 ± 49 Ω × cm 2 and fluorescein permeability below 1.95 × 10 -7 cm/s. E6-derived BMECs maintained TEER above 1000 Ω × cm 2 for a minimum of 8 days and showed no statistical difference in efflux transporter activity compared to BMECs differentiated by conventional means. The method was also found to support long-term stability of BMECs harboring biallelic PARK2 mutations associated

  4. Differentiation of a bipotential glial progenitor cell in a single cell microculture.

    Science.gov (United States)

    Temple, S; Raff, M C

    Although it is known that most cells of the vertebrate central nervous system (CNS) are derived from the neuroepithelial cells of the neural tube, the factors determining whether an individual neuroepithelial cell develops into a particular type of neurone or glial cell remain unknown. A promising model for studying this problem is the bipotential glial progenitor cell in the developing rat optic nerve; this cell differentiates into a particular type of astrocyte (a type-2 astrocyte) if cultured in 10% fetal calf serum (FCS) and into an oligodendrocyte if cultured in serum-free medium. As the oligodendrocyte-type-2 astrocyte (0-2A) progenitor cell can differentiate along either glial pathway in neurone-free cultures, living axons clearly are not required for its differentiation, at least in vitro. However, the studies on 0-2A progenitor cells were carried out in bulk cultures of optic nerve, and so it was possible that other cell-cell interactions were required for differentiation in culture. We show here that 0-2A progenitor cells can differentiate into type-2 astrocytes or oligodendrocytes when grown as isolated cells in microculture, indicating that differentiation along either glial pathway in vitro does not require signals from other CNS cells, apart from the signals provided by components of the culture medium. We also show that single 0-2A progenitor cells can differentiate along either pathway without dividing, supporting our previous studies using 3H-thymidine and suggesting that DNA replication is not required for these cells to choose between the two differentiation programmes.

  5. Endothelial Progenitor Cell Fraction Contained in Bone Marrow-Derived Mesenchymal Stem Cell Populations Impairs Osteogenic Differentiation

    Directory of Open Access Journals (Sweden)

    Fabian Duttenhoefer

    2015-01-01

    Full Text Available In bone tissue engineering (TE endothelial cell-osteoblast cocultures are known to induce synergies of cell differentiation and activity. Bone marrow mononucleated cells (BMCs are a rich source of mesenchymal stem cells (MSCs able to develop an osteogenic phenotype. Endothelial progenitor cells (EPCs are also present within BMC. In this study we investigate the effect of EPCs present in the BMC population on MSCs osteogenic differentiation. Human BMCs were isolated and separated into two populations. The MSC population was selected through plastic adhesion capacity. EPCs (CD34+ and CD133+ were removed from the BMC population and the resulting population was named depleted MSCs. Both populations were cultured over 28 days in osteogenic medium (Dex+ or medium containing platelet lysate (PL. MSC population grew faster than depleted MSCs in both media, and PL containing medium accelerated the proliferation for both populations. Cell differentiation was much higher in Dex+ medium in both cases. Real-time RT-PCR revealed upregulation of osteogenic marker genes in depleted MSCs. Higher values of ALP activity and matrix mineralization analyses confirmed these results. Our study advocates that absence of EPCs in the MSC population enables higher osteogenic gene expression and matrix mineralization and therefore may lead to advanced bone neoformation necessary for TE constructs.

  6. Acceleration of diabetic wound healing with adipose-derived stem cells, endothelial-differentiated stem cells, and topical conditioned medium therapy in a swine model.

    Science.gov (United States)

    Irons, Robin F; Cahill, Kevin W; Rattigan, Deviney A; Marcotte, Joseph H; Fromer, Marc W; Chang, Shaohua; Zhang, Ping; Behling, Eric M; Behling, Kathryn C; Caputo, Francis J

    2018-05-09

    The purpose of our study was to investigate the effect of adipose-derived stem cells (ASCs), endothelial-differentiated ASCs (EC/ASCs), and various conditioned media (CM) on wound healing in a diabetic swine model. We hypothesized that ASC-based therapies would accelerate wound healing. Diabetes was induced in four Yorkshire swine through intravenous injection of streptozotocin. ASCs were harvested from flank fat and cultured in either M199 or EGM-2 medium. A duplicate series of seven full-thickness dorsal wounds were surgically created on each swine. The wounds in the cellular treatment group underwent injection of low-dose or high-dose ASCs or EC/ASCs on day 0, with a repeat injection of one half of the initial dose on day 15. Wounds assigned to the topical CM therapy were covered with 2 mL of either serum-free M199 primed by ASCs or human umbilical vein endothelial cells every 3 days. Wounds were assessed at day 0, 10, 15, 20, and 28. The swine were sacrificed on day 28. ImageJ software was used to evaluate the percentage of wound healing. The wounded skin underwent histologic, reverse transcription polymerase chain reaction, and enzyme-linked immunosorbent assay examinations to evaluate markers of angiogenesis and inflammation. We found an increase in the percentage of wound closure rates in cell-based treatments and topical therapies at various points compared with the untreated control wounds (P swine model. Enhanced angiogenesis and immunomodulation might be key contributors to this process. The purpose of the present study was to translate the known beneficial effects of adipose-derived stem cells and associated conditioned medium therapy on diabetic wound healing to a large animal model. We demonstrated that stem cell and conditioned medium therapy significantly accelerate gross wound healing in diabetic swine, with data suggesting this might result from a decreased inflammatory response and increased angiogenesis. Copyright © 2018 Society for

  7. Isolation and in vitro culture of trypanosomes from Leptodactylus ocellatus from the Atlantic Forest in a new experimental culture medium.

    Science.gov (United States)

    Lemos, M; Souza, C S F; da Costa, S C Gonçalves; Souto-Padrón, T; D'Agosto, M

    2013-02-01

    The purpose of this study was to verify the in vitro development of Trypanosoma sp. isolated from Leptodactylus ocellatus frogs under a new protocol using a biphasic medium composed of Novy, McNeal, and Nicolle (NNN) blood agar medium as a solid phase and liver infusion, brain heart infusion, and tryptose (LIBHIT) medium as a liquid phase. Blood forms, collected by cardiac puncture or after the maceration of different organs, were inoculated in culture tubes containing the biphasic medium composed by NNN and LIBHIT. Trypanosomes were observed 4 days postinoculation; most bloodstream trypomastigotes had differentiated into epimastigotes and amastigotes by this time. Trypomastigotes were again observed in older cultures (7 days). Parasites were successfully subcultured for 8 mo in this medium and successfully cryopreserved. The present study provides a new protocol medium for the isolation and culture of anuran trypanosomes.

  8. Distinction of Gram-positive and -negative bacteria using a colorimetric microbial viability assay based on the reduction of water-soluble tetrazolium salts with a selection medium.

    Science.gov (United States)

    Tsukatani, Tadayuki; Suenaga, Hikaru; Higuchi, Tomoko; Shiga, Masanobu; Noguchi, Katsuya; Matsumoto, Kiyoshi

    2011-01-01

    Bacteria are fundamentally divided into two groups: Gram-positive and Gram-negative. Although the Gram stain and other techniques can be used to differentiate these groups, some issues exist with traditional approaches. In this study, we developed a method for differentiating Gram-positive and -negative bacteria using a colorimetric microbial viability assay based on the reduction of the tetrazolium salt {2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt} (WST-8) via 2-methyl-1,4-napthoquinone with a selection medium. We optimized the composition of the selection medium to allow the growth of Gram-negative bacteria while inhibiting the growth of Gram-positive bacteria. When the colorimetric viability assay was carried out in a selection medium containing 0.5µg/ml crystal violet, 5.0 µg/ml daptomycin, and 5.0µg/ml vancomycin, the reduction in WST-8 by Gram-positive bacteria was inhibited. On the other hand, Gram-negative bacteria produced WST-8-formazan in the selection medium. The proposed method was also applied to determine the Gram staining characteristics of bacteria isolated from various foodstuffs. There was good agreement between the results obtained using the present method and those obtained using a conventional staining method. These results suggest that the WST-8 colorimetric assay with selection medium is a useful technique for accurately differentiating Gram-positive and -negative bacteria.

  9. Determination of the molecular structure via the medium energy electrons (500 eV-1,5 KeV) Ar, N2, Co e HCl

    International Nuclear Information System (INIS)

    Nogueira, J.C.

    1977-01-01

    Elastic Differential and Total Differential Cross Sections are measured for electron collision in medium-energy range (500 eV - 1,5 KeV) with argon, nitrogen, carbon monoxide and hydrogen chloride, all in their electronic ground state. Theoretical calculation for the Elastic Differential Cross Sections by atoms were done employing Hartree-Fock-Clementy wave function, and making use of Partial Wave and WKBJ Methods. Exchange effect is included in the case of argon. Independent Atom Model, Half Molecule Model and a new model, the Ionic Model were utilized for the molecular calculations. The Ionic Model is suggested for the interaction between HCl and electrons. Inelastic Differential Cross Section were also computed, making use of the First Born Approximation and Hartree-Fock-Clementi wave function. It is also demonstrated, for the first time, that medium energy electrons (500 eV - 1,5 Kev) can be used to determine molecular structure parameters, in gas phase [pt

  10. Differentiation of Yersinia enterocolitica biotype 1A from pathogenic Yersinia enterocolitica biotypes by detection of β-glucosidase activity: comparison of two chromogenic culture media and Vitek2.

    Science.gov (United States)

    Karhukorpi, Jari; Päivänurmi, Marjut

    2014-01-01

    Aesculin hydrolysis (ESC) is one of the key reactions in differentiating pathogenic Yersinia enterocolitica biotypes 1B, 2, 3, 4 and 5 from the less-pathogenic biotype 1A. Because the ESC reaction is caused by β-glucosidase (βGLU) activity of the bacteria, we studied whether two commonly used methods (BBL CHROMagar Orientation and Vitek2 Gram-negative identification card) could be used in assessing βGLU activity of 74 Yersinia strains. Both methods were sensitive (100 % and 97 %) and specific (100 % and 100 %) in differentiating βGLU-positive YE BT1A from βGLU-negative Y. enterocolitica biotypes. For a subset of strains (n = 69), a new selective CHROMagar Yersinia showed excellent agreement with the strains' βGLU activity. Thus all the methods evaluated in this study may be used to differentiate between YE BT1A and other Y. enterocolitica biotypes.

  11. Pulsed magnetic therapy increases osteogenic differentiation of mesenchymal stem cells only if they are pre-committed.

    Science.gov (United States)

    Ferroni, Letizia; Tocco, Ilaria; De Pieri, Andrea; Menarin, Martina; Fermi, Enrico; Piattelli, Adriano; Gardin, Chiara; Zavan, Barbara

    2016-05-01

    Pulsed electromagnetic field (PEMF) therapy has been documented to be an effective, non-invasive, safe treatment method for a variety of clinical conditions, especially in settings of recalcitrant healing. The underlying mechanisms on the different biological components of tissue regeneration are still to be elucidated. The aim of the present study was to characterize the effects of extremely low frequency (ELF)-PEMFs on commitment of mesenchymal stem cell (MSCs) culture system, through the determination of gene expression pattern and cellular morphology. Human MSCs derived from adipose tissue (ADSCs) were cultured in presence of adipogenic, osteogenic, neural, or glial differentiative medium and basal medium, then exposed to ELF-PEMFs daily stimulation for 21days. Control cultures were performed without ELF-PEMFs stimulation for all cell populations. Effects on commitment were evaluated after 21days of cultures. The results suggested ELF-PEMFs does not influence ADSCs commitment and does not promote adipogenic, osteogenic, neural or glial differentiation. However, ELF-PEMFs treatment on ADSCs cultured in osteogenic differentiative medium markedly increased osteogenesis. We concluded that PEMFs affect the osteogenic differentiation of ADSCs only if they are pre-commitment and that this therapy can be an appropriate candidate for treatment of conditions requiring an acceleration of repairing process. Copyright © 2016 Elsevier Inc. All rights reserved.

  12. 'That proves my point': How mediums reconstrue disconfirmation in medium-sitter interactions.

    Science.gov (United States)

    Enoksen, Anette Einan; Dickerson, Paul

    2018-04-01

    Previous research has examined how the talk of mediums attends to the epistemological status of their readings. Such work has identified that mediums frequently use question-framed propositions that are typically confirmed by the sitter, thereby conferring epistemological status on the medium. This study seeks to investigate what happens when the sitter disconfirms the propositions of the medium. The study focuses on the ways in which such disconfirmation can be responded to such that it is reconstrued as evidence of the psychic nature of the medium's reading. Televised demonstrations of psychic readings involving British and US mediums and their sitters are analysed. The results suggest that mediums rework disconfirmation as proof in several ways: first, by emphasizing the different access that sitter and medium have to knowledge (e.g., about the future); second, as evidence that the medium has access to the actual voice of the deceased (and may therefore mishear what the deceased has said to them); and third, as revealing an important truth that has hitherto been concealed from the sitter. The implications of these findings are considered for cases where speakers bring different and potentially competing, epistemological resources to an interaction. © 2018 The British Psychological Society.

  13. Detection of distribution of avian influenza H5N1 virus by immunohistochemistry, chromogenic in situ hybridization and real-time PCR techniques in experimentally infected chickens.

    Science.gov (United States)

    Chamnanpood, Chanpen; Sanguansermsri, Donruedee; Pongcharoen, Sutatip; Sanguansermsri, Phanchana

    2011-03-01

    Ten specific pathogen free (SPF) chickens were inoculated intranasally with avian influenza virus subtype H5N1. Evaluation revealed distribution of the virus in twelve organs: liver, intestine, bursa, lung, trachea, thymus, heart, pancreas, brain, spleen, kidney, and esophagus. Immunohistochemistry (IHC), chromogenic in situ hybridization (CISH), and real-time polymerase chain reaction (PCR) were developed and compared for detection of the virus from the organs. The distribution of avian influenza H5N1 in chickens varied by animal and detecting technique. The heart, kidneys, intestines, lungs, and pancreas were positive with all three techniques, while the others varied by techique. The three techniques can be used to detect avian influenza effectively, but the pros and cons of each technique need to be determined. The decision of which technique to use depends on the objective of the examination, budget, type and quality of samples, laboratory facilities and technician skills.

  14. Nonlinear radiative peristaltic flow of hydromagnetic fluid through porous medium

    Directory of Open Access Journals (Sweden)

    Q. Hussain

    2018-06-01

    Full Text Available The radiative heat and mass transfer in wall induced flow of hydromagnetic fluid through porous medium in an asymmetric channel is analyzed. The fluid viscosity is considered temperature dependent. In the theory of peristalsis, the radiation effects are either ignored or taken as linear approximation of radiative heat flux. Such approximation is only possible when there is sufficiently small temperature differences in the flow field; however, nonlinear radiation effects are valid for large temperature differences as well (the new feature added in the present study. Mathematical modeling of the problems include the complicated system of highly nonlinear differential equations. Semi-analytical solutions are established in the wave reference frame. Results are displayed graphically and discussed in detail for the variation of various physical parameters with the special attention to viscosity, radiation, and temperature ratio parameters. Keywords: Nonlinear thermal radiation, Variable viscosity, Porous medium, Soret and Dufour effects, Peristalsis

  15. Neural stem cells was induced to differentiate into cholinergic neurons in vitro

    International Nuclear Information System (INIS)

    Chang Yan; Xu Yilong; Pan Jingkun; Tian Lei; Gao Yuhong; Guo Shuilong

    2004-01-01

    The cholinergic-inducing effect of BMP4 on isolated and cultivated rat's cerebral neural stem cells (NSCs) was examined. NSCs which were isolated from two month's old rat's brain region like hippocampus and striatum were cultivated in a medium containing EGF and bFGF, and were identified with morphological character by microscope and nestin immunocytochemistry test. After 24 hours, half NSCs were cultivated with a BMP4-added medium as a experimental group instead of the primary medium, while the an other half NSCs being cultivated with the primary medium as a control group. After 8 days the expression of choline acetyltransferase (ChAT) of the cultivated cells was observated by indirect immunofluorescence test. Results showed that more positive cells were found in the experimental group, and the fluorescence intensity were stronger; while less positive cells were found in the control group, and the fluorescence intensity was weaker. The differentiational efficiency of the NSCs was examined by FITC-labelled Flow Cytometry. The results showed that about 16% cells of the experimental group appeared ChAT-positive, while that of control group only 7%. So BMP4 may have the function of inducing NSCs to differentiate into neurons with cholinergic characteristic. (authors)

  16. Comparison of Fluorescence In Situ Hybridization and Chromogenic In Situ Hybridization for Low and High Throughput HER2 Genetic Testing

    Science.gov (United States)

    Poulsen, Tim S.; Espersen, Maiken L. M.; Kofoed, Vibeke; Dabetic, Tanja; Høgdall, Estrid; Balslev, Eva

    2013-01-01

    The purpose was to evaluate and compare 5 different HER2 genetic assays with different characteristics that could affect the performance to analyze the human epidermal growth factor 2 (HER2) gene copy number under low and high throughput conditions. The study included 108 tissue samples from breast cancer patients with HER2 immunohistochemistry (IHC) results scored as 0/1+, 2+, and 3+. HER2 genetic status was analysed using chromogenic in situ hybridization (CISH) and fluorescence in situ hybridization (FISH). Scoring results were documented through digital image analysis. The cancer region of interest was identified from a serial H&E stained slide following tissue cores were transferred to a tissue microarrays (TMA). When using TMA in a routine flow, all patients will be tested for HER2 status with IHC followed by CISH or FISH, thereby providing individual HER2 results. In conclusion, our results show that the differences between the HER2 genetic assays do not have an effect on the analytic performance and the CISH technology is superior to high throughput HER2 genetic testing due to scanning speed, while the IQ-FISH may still be a choice for fast low throughput HER2 genetic testing. PMID:24383005

  17. Evaluation of HER2 gene amplification in invasive breast cancer using a dual-color chromogenic in situ hybridization (dual CISH).

    Science.gov (United States)

    Kato, Nobuaki; Itoh, Hitoshi; Serizawa, Akihiko; Hatanaka, Yutaka; Umemura, Shinobu; Osamura, R Yoshiyuki

    2010-07-01

    Fluorescence in situ hybridization (FISH) assay is considered the 'gold standard' for evaluation of HER2/neu (HER2) gene status, however, it is difficult to recognize morphologic features of tumors using fluorescence microscopy. Thus, chromogenic in situ hybridization (CISH) has been proposed as an alternative method to evaluate HER2 gene amplification. Here, we examined the dual color CISH (dual CISH) method which provides information regarding the copy number of the HER2 gene and chromosome 17 centromere from a single slide. We examined 40 cases of invasive ductal carcinomas of the breast that were resected surgically. HER2 gene status was assessed with FISH (Abbott) and dual CISH (Dako). HER2 gene amplification status was classified according to the guidelines of the American Society of Clinical Oncology and College of American Pathologists (ASCO/CAP). Comparison of the cut-off values for HER2/chromosome 17 centromere copy number ratio obtained by dual CISH and FISH showed that there was almost perfect agreement between two methods (Kappa coefficient 0.96). The results of the two commercial products were almost consistent for evaluation of HER2 gene counts on the sections. The current study proved that dual CISH is comparable with FISH for evaluating HER2 gene status.

  18. Bright-field in situ hybridization for HER2 gene amplification in breast cancer using tissue microarrays: correlation between chromogenic (CISH) and automated silver-enhanced (SISH) methods with patient outcome.

    Science.gov (United States)

    Francis, Glenn D; Jones, Mark A; Beadle, Geoffrey F; Stein, Sandra R

    2009-06-01

    HER2 gene amplification or overexpression occurs in 15% to 25% of breast cancers and has implications for treatment and prognosis. The most commonly used methods for HER2 testing are fluorescence in situ hybridization (FISH) and immunohistochemistry. FISH is considered to be the reference standard and more accurately predicts response to trastuzumab, but is technically demanding, expensive, and requires specialized equipment. In situ hybridization is required to be eligible for adjuvant treatment with trastuzumab in Australia. Bright-field in situ hybridization is an alternative to FISH and uses a combination of in situ methodology and a peroxidase-mediated chromogenic substrate such as diaminobenzidine [chromogenic in situ hybridization (CISH)] or multimer technology coupled with enzyme metallography [silver-enhanced in situ hybridization (SISH)] to create a marker visible under bright-field microscopy. CISH was introduced into diagnostic testing in Australia in October 2006. SISH methodology is a more recent introduction into the testing repertoire. An evaluation of CISH and SISH performance to assess patient outcome were performed using tissue microarrays. Tissue microarrays were constructed in duplicate using material from 593 patients with invasive breast carcinoma and assessed using CISH and SISH. Gene amplification was assessed using the American Society of Clinical Oncology/College of American Pathologists guideline and Australian HER2 Advisory Board criteria (single probe: diploid, 1 to 2.5 copies/nucleus; polysomy >2.5 to 4 copies/nucleus; equivocal, >4 to 6 copies/nucleus; low-level amplification, >6 to 10 copies/nucleus and high-level amplification >10 copies/nucleus; dual probe HER2/CHR17 ratio: nonamplified 2.2). Results were informative for 337 tissue cores comprising 230 patient samples. Concordance rates were 96% for HER2 single probe CISH and SISH and 95.5% for single probe CISH and dual probe HER2/CHR17 SISH. Both bright-field methods correlated

  19. Effects of trichostatins on differentiation of murine erythroleukemia cells

    International Nuclear Information System (INIS)

    Yoshida, M.; Nomura, S.; Beppu, T.

    1987-01-01

    The fungistatic antibiotics trichostatins (TS) A and C were isolated from culture broth of Streptomyces platensis No. 145 and were found to be potent inducers of differentiation in murine erythroleukemia (Friend and RV133) cells at concentrations of 1.5 X 10(-8) M for TSA and 5 X 10(-7) M for TSC. Differentiation induced by TS was cooperatively enhanced by UV irradiation but not by treatment with dimethyl sulfoxide. This enhanced activity was completely inhibited by adding cycloheximide to the culture medium 2 h after exposure to TS, suggesting that TS are dimethyl sulfoxide-type inducers of erythroid differentiation. No inhibitory effect of TS was observed on macromolecular synthesis in cultured cells

  20. Impact of culture medium on maturation of bone marrow-derived murine dendritic cells via the aryl hydrocarbon receptor.

    Science.gov (United States)

    Ilchmann, Anne; Krause, Maren; Heilmann, Monika; Burgdorf, Sven; Vieths, Stefan; Toda, Masako

    2012-05-01

    The aryl hydrocarbon receptor (AhR) plays a role in modulating dendritic cell (DC) immunity. Iscove's modified Dulbecco's medium (IMDM) contains higher amounts of AhR ligands than RPMI1640 medium. Here, we examined the influence of AhR ligand-containing medium on the maturation and T-cell stimulatory capacity of bone marrow-derived murine dendritic cells (BMDCs). BMDCs generated in IMDM (BMDCs/IMDM) expressed higher levels of co-stimulatory and MHC class II molecules, and lower levels of pattern-recognition receptors, especially toll-like receptor (TLR) 2, TLR4, and scavenger receptor class A (SR-A), compared to BMDCs generated in RPMI1640 medium (BMDCs/RPMI). Cytokine responses against ligands of TLRs and antigen uptake mediated by SR-A were remarkably reduced in BMDCs/IMDM, whereas the T-cell stimulatory capacity of the cells was enhanced, compared to BMDCs/RPMI. The enhanced maturation of BMDCs/IMDM was attenuated in the presence of an AhR antagonist, indicating involvement of AhR in the maturation. Interestingly, BMDCs/IMDM induced Th2 and Th17 differentiation at low and high concentrations of antigen respectively, when co-cultured with CD4(+) T-cells from antigen-specific T-cell receptor transgenic mice. In contrast, BMDCs/RPMI induced Th1 differentiation predominantly in the co-culture. Taken together, optimal selection of medium seems necessary when studying BMDCs, depending on the target receptors on the cell surface of DCs and type of helper T-cells for the co-culture. Copyright © 2012 Elsevier Ltd. All rights reserved.

  1. Giant arachnoid granulation: differential diagnosis of acute headache

    International Nuclear Information System (INIS)

    Peters, S.A.; Heyer, C.M.; Frombach, E.

    2007-01-01

    Full text: We present a case of intense, rapidly evolving headache clinically mimicking meningitis, subarachnoid haemorrhage or venous sinus thrombosis. Clinical examination, standard blood work and central nervous system studies were non-contributory and effectively ruled out these diagnoses. Cranial multidetector CT studies before and after application of intravenous contrast medium performed prior to lumbar tap disclosed a non-enhancing ovoid mass filling the superior sagittal sinus. This lesion posed a differential to venous sinus thrombosis, but ultimately fulfilled the criteria of a giant arachnoid granulation. The imaging characteristics and differential diagnosis of giant arachnoid granulations are discussed

  2. Dual-colour chromogenic in-situ hybridization is a potential alternative to fluorescence in-situ hybridization in HER2 testing.

    Science.gov (United States)

    Hwang, Cheng-Cheng; Pintye, Mariann; Chang, Liang-Che; Chen, Huang-Yang; Yeh, Kun-Yan; Chein, Hui-Ping; Lee, Nin; Chen, Jim-Ray

    2011-11-01

    Dual-colour chromogenic in-situ hybridization (dc-CISH) is an emerging methodology for characterizing genomic alterations. This study was aimed at evaluating the performance of a dc-CISH kit (ZytoVision) in determining human epidermal growth factor receptor 2 (HER2) status in breast cancer. Two hundred and twenty-eight invasive breast carcinomas arranged in tissue microarrays were analysed in parallel with dc-CISH, fluorescence in-situ hybridization (FISH), and immunohistochemistry. Of 227 tumours with available FISH and dc-CISH results, HER2 amplification and non-amplification were detected in 49 (21.6%) and 178 (78.4%) tumours, respectively, by both assays. The concordance between dc-CISH and FISH results showed 100% agreement (κ-coefficient=1.00). Immunohistochemically, 162 (71%), 25 (11.0%) and 41 (18%) tumours were scored 0/1+, 2+, and 3+, respectively. The corresponding results with both FISH and dc-CISH demonstrated HER2 amplification in two (3.2%), nine (36%) and 38 (93%) tumours, respectively. Complete consensus among these three methods was observed in 197 cases, representing 98% of all 3+ and 0/1+ tumours (κ-coefficient=0.92). Confirmatory testing of 25 2+ tumours showed complete consensus between FISH and dc-CISH. dc-CISH is a promising alternative to FISH in HER2 testing, and the single-institute incidence of HER2 amplification in breast cancer in Taiwan is 21.2%. © 2011 Blackwell Publishing Limited.

  3. An automatic flow injection analysis procedure for photometric determination of ethanol in red wine without using a chromogenic reagent.

    Science.gov (United States)

    Borges, Sivanildo S; Frizzarin, Rejane M; Reis, Boaventura F

    2006-05-01

    An automatic reagentless photometric procedure for the determination of ethanol in red wine is described. The procedure was based on a falling drop system that was implemented by employing a flow injection analysis manifold. The detection system comprised an infrared LED and a phototransistor. The experimental arrangement was designed to ensure that the wine drop grew between these devices, thus causing a decrease in the intensity of the radiation beam coming from the LED. Since ethanol content affected the size of the wine drop this feature was exploited to develop an analytical procedure for the photometric determination of ethanol in red wine without using a chromogenic reagent. In an attempt to prove the usefulness of the proposed procedure, a set of red wines were analysed. No significant difference between our results and those obtained with a reference method was observed at the 95% confidence level. Other advantages of our method were a linear response ranging from 0.17 up to 5.14 mol L(-1) (1.0 up to 30.0%) ethanol (R = 0.999); a limit of detection of 0.05 mol L(-1) (0.3%) ethanol; a relative standard deviation of 2.5% (n = 10) using typical wine sample containing 2.14 mol L(-1) (12.5%) ethanol; and a sampling rate of 50 determinations per hour.

  4. Identification of MYCN gene amplification in neuroblastoma using chromogenic in situ hybridization (CISH): an alternative and practical method.

    Science.gov (United States)

    Bhargava, Rohit; Oppenheimer, Orit; Gerald, William; Jhanwar, Suresh C; Chen, Beiyun

    2005-06-01

    Chromogenic in situ hybridization (CISH) is a recently developed technique, which utilizes the general principles of in situ hybridization and a detection system similar to immunohistochemistry. To assess the utility of CISH for analysis of MYCN gene amplification, we compared this assay with established diagnostic assays such as Southern blot analysis (SB) and fluorescent in situ hybridization (FISH). CISH was performed on 67 cases of neuroblastoma using tissue microarray (65 cases) and whole tissue sections (2 cases). Unequivocal, high-level amplification (> or =10 gene copies per tumor nucleus) was identified in 19 of 67 (28.4%) tumors. Two (3%) tumors showed low-level amplification (6-9 gene copies per tumor nucleus). No amplification was seen in 46 of 67 (68.6%) tumors. SB data were available in 44 tumors. Forty-one of the 44 tumors (93%) showed concordant results between CISH and SB. Three tumors showed MYCN amplification by CISH but no amplification by SB, most likely due to dilution effect of nonneoplastic tissue in the test samples. Two of these three tumors also showed MYCN amplification by FISH, and the third tumor was not analyzed by FISH. FISH data were available in total of 30 tumors. All 30 tumors showed concordant results between CISH and FISH for classifying a tumor as MYCN amplified or not amplified. We conclude that CISH is an accurate method for determining MYCN gene amplification, with added advantages that make it a more practically useful method.

  5. Naringin promotes osteogenic differentiation of bone marrow stromal cells by up-regulating Foxc2 expression via the IHH signaling pathway.

    Science.gov (United States)

    Lin, Fei-Xiang; Du, Shi-Xin; Liu, De-Zhong; Hu, Qin-Xiao; Yu, Guo-Yong; Wu, Chu-Cheng; Zheng, Gui-Zhou; Xie, Da; Li, Xue-Dong; Chang, Bo

    2016-01-01

    Naringin is an active compound extracted from Rhizoma Drynariae, and studies have revealed that naringin can promote proliferation and osteogenic differentiation of bone marrow stromal cells (BMSCs). In this study, we explored whether naringin could promote osteogenic differentiation of BMSCs by upregulating Foxc2 expression via the Indian hedgehog (IHH) signaling pathway. BMSCs were cultured in basal medium, basal medium with naringin, osteogenic induction medium, osteogenic induction medium with naringin and osteogenic induction medium with naringin in the presence of the IHH inhibitor cyclopamine (CPE). We examined cell proliferation by using a WST-8 assay, and differentiation by Alizarin Red S staining (for mineralization) and alkaline phosphatase (ALP) activity. In addition, we detected core-binding factor α1 (Cbfα1), osteocalcin (OCN), bone sialoprotein (BSP), peroxisome proliferation-activated receptor gamma 2 (PPARγ2) and Foxc2 expression by using RT-PCR. We also determined Foxc2 and IHH protein levels by western blotting. Naringin increased the mineralization of BMSCs, as shown by Alizarin red S assays, and induced ALP activity. In addition, naringin significantly increased the mRNA levels of Foxc2, Cbfα1, OCN, and BSP, while decreasing PPARγ2 mRNA levels. Furthermore, the IHH inhibitor CPE inhibited the osteogenesis-potentiating effects of naringin. Naringin increased Foxc2 and stimulated the activation of IHH, as evidenced by increased expression of proteins that were inhibited by CPE. Our findings indicate that naringin promotes osteogenic differentiation of BMSCs by up-regulating Foxc2 expression via the IHH signaling pathway.

  6. Human endometrial stromal stem cells differentiate into megakaryocytes with the ability to produce functional platelets.

    Directory of Open Access Journals (Sweden)

    Jinju Wang

    Full Text Available Human endometrium is a high dynamic tissue that contains endometrial stromal stem cells (hESSCs. The hESSCs have been differentiated into a number of cell lineages. However, differentiation of hESSCs into megakaryocytes (MKs has not yet been investigated. The aim of this study was to investigate the feasibility of MK generation from hESSCs and subsequent production of functional platelets (PLTs. In our study, hESSCs were cultured from endometrial stromal cells as confirmed by positive stromal cell specific markers (CD90 and CD29 and negative hematopoietic stem cell markers (CD45 and CD34 expression. Then, hESSCs were differentiated in a medium supplemented with thrombopoietin (TPO for 18 days. The MK differentiation was analyzed by flow cytometry and confocal microscopy. The differentiation medium was collected for PLT production analysis by flow cytometry, transmission electron microscopy and functional measurements. Our results show: 1 MKs were successfully generated from hESSCs as identified by expression of specific markers (CD41a: 1 ± 0.09% and 39 ± 3.0%; CD42b: 1.2 ± 0.06% and 28 ± 2.0%, control vs. differentiation accompanied with reduction of pluripotent transcription factors (Oct4 and Sox2 expression; 2 The level of PLTs in the differentiation medium was 16 ± 1 number/µl as determined by size (2-4 µm and CD41a expression (CD41a: 1 ± 0.4% and 90±2.0%, control vs. differentiation; 3 Generated PLTs were functional as evidenced by the up-regulation of CD62p expression and fibrinogen binding following thrombin stimulation; 4 Released PLTs showed similar ultra-structure characteristics (alpha granules, vacuoles and dense tubular system as PLTs from peripheral blood determined by electron microscopic analysis. Data demonstrate the feasibility of generating MKs from hESSCs, and that the generated MKs release functional PLTs. Therefore, hESSCs could be a potential new stem cell source for in vitro MK/PLT production.

  7. Control of exoenzyme production, motility and cell differentiation in Serratia liquefaciens

    DEFF Research Database (Denmark)

    Givskov, Michael Christian; Eberl, Leo; Molin, Søren

    1997-01-01

    Serratia liquefaciens secretes a broad spectrum of hydrolytic enzymes to the surrounding medium and possesses the ability to differentiate into specialized swarmer cells capable of rapid surface motility. Control of exoenzyme production and swarming motility is governed by similar regulatory...

  8. In vitro differentiation of neural cells from human adipose tissue derived stromal cells.

    Science.gov (United States)

    Dave, Shruti D; Patel, Chetan N; Vanikar, Aruna V; Trivedi, Hargovind L

    2018-01-01

    Stem cells, including neural stem cells (NSCs), are endowed with self-renewal capability and hence hold great opportunity for the institution of replacement/protective therapy. We propose a method for in vitro generation of stromal cells from human adipose tissue and their differentiation into neural cells. Ten grams of donor adipose tissue was surgically resected from the abdominal wall of the human donor after the participants' informed consents. The resected adipose tissue was minced and incubated for 1 hour in the presence of an enzyme (collagenase-type I) at 37 0 C followed by its centrifugation. After centrifugation, the supernatant and pellets were separated and cultured in a medium for proliferation at 37 0 C with 5% CO2 for 9-10 days in separate tissue culture dishes for generation of mesenchymal stromal cells (MSC). At the end of the culture, MSC were harvested and analyzed. The harvested MSC were subjected for further culture for their differentiation into neural cells for 5-7 days using differentiation medium mainly comprising of neurobasal medium. At the end of the procedure, culture cells were isolated and studied for expression of transcriptional factor proteins: orthodenticle homolog-2 (OTX-2), beta-III-tubulin (β3-Tubulin), glial-fibrillary acid protein (GFAP) and synaptophysin-β2. In total, 50 neural cells-lines were generated. In vitro generated MSC differentiated neural cells' mean quantum was 5.4 ± 6.9 ml with the mean cell count being, 5.27 ± 2.65 × 10 3/ μl. All of them showed the presence of OTX-2, β3-Tubulin, GFAP, synaptophysin-β2. Neural cells can be differentiated in vitro from MSC safely and effectively. In vitro generated neural cells represent a potential therapy for recovery from spinal cord injuries and neurodegenerative disease.

  9. Differentiation of Spermatogonia Stem Cells into Functional Mature Neurons Characterized with Differential Gene Expression.

    Science.gov (United States)

    Bojnordi, Maryam Nazm; Azizi, Hossein; Skutella, Thomas; Movahedin, Mansoureh; Pourabdolhossein, Fereshteh; Shojaei, Amir; Hamidabadi, Hatef Ghasemi

    2017-09-01

    Transplantation of embryonic stem cells (ESCs) is a promising therapeutic approach for the treatment of neurodegenerative diseases. However, ESCs are not usable clinically due to immunological and ethical limitations. The identification of an alternative safe cell source opens novel options via autologous transplantation in neuro-regeneration circumventing these problems. Here, we examined the neurogenic capacity of embryonic stem-like cells (ES-like cells) derived from the testis using neural growth factor inducers and utilized them to generate functional mature neurons. The neuronal differentiation of ES-like cells is induced in three stages. Stage 1 is related to embryoid body (EB) formation. To induce neuroprogenitor cells, EBs were cultured in the presence of retinoic acid, N 2 supplement and fibroblast growth factor followed by culturing in a neurobasal medium containing B 27 , N 2 supplements for additional 10 days, to allow the maturation and development of neuronal progenitor cells. The neurogenic differentiation was confirmed by immunostaining for markers of mature neurons. The differentiated neurons were positive for Tuj1 and Tau1. Real-time PCR dates indicated the expression of Nestin and Neuro D (neuroprogenitor markers) in induced cells at the second stage of the differentiation protocol. The differentiated mature neurons exhibited the specific neuron markers Map2 and β-tubulin. The functional maturity of neurons was confirmed by an electrophysiological analysis of passive and active neural membrane properties. These findings indicated a differentiation capacity of ES-like cells derived from the testis to functionally mature neurons, which proposes them as a novel cell source for neuroregenerative medicine.

  10. Simulation of Reynolds number influence on heat exchange in turbulent flow of medium slurry

    Science.gov (United States)

    Bartosik, A.

    2016-10-01

    The paper deals with the numerical simulation of mass and heat exchange in turbulent flow of solid-liquid mixture in the range of averaged solid particle diameter from 0.10mm to 0.80mm, named further as the medium slurry. Physical model assumes that dispersed phase is fully suspended and a turbulent flow is hydro-dynamically, and thermally developed in a straight horizontal pipeline. Taking into account the aforementioned assumptions the slurry is treated as a single-phase flow with increased density, while viscosity is equals to a carrier liquid viscosity. The mathematical model constitutes time averaged momentum equation in which the turbulent stress tensor was designated using a two-equation turbulence model, which makes use of the Boussinesq eddy-viscosity hypothesis. Turbulence damping function in the turbulence model was especially designed for the medium slurry. In addition, an energy equation has been used in which a convective term was determined from the energy balance acting on a unit pipe length, assuming linear changes of temperature in main flow direction. Finally, the mathematical model of non-isothermal medium slurry flow comprises four partial differential equations, namely momentum and energy equations, equations of kinetic energy of turbulence and its dissipation rate. Four partial differential equations were solved by a finite difference scheme using own computer code. The objective of the paper is to examine the influence of Reynolds number on temperature profiles and Nusselt number in turbulent flow of medium slurry in the range of solids concentration from 0% to 30% by volume. The effect of influential factors on heat transfer between the pipe and slurry is analysed. The paper demonstrates substantial impact of Reynolds number and solids volume fraction on the Nusselt number. The results of numerical simulation are reviewed.

  11. The influence of serum-free culture conditions on skeletal muscle differentiation in a tissue-engineered model

    NARCIS (Netherlands)

    Gawlitta, D.; Boonen, K.J.M.; Oomens, C.W.J.; Baaijens, F.P.T.; Bouten, C.V.C.

    2008-01-01

    The influence of differentiation medium (DM) components on C2C12 murine myoblast differentiation has only been studied in monolayer cultures. Serum-free formulations have been applied that omit the use of sera with unknown composition. The goal of the present study was to compare the influence of

  12. Role of human amnion-derived mesenchymal stem cells in promoting osteogenic differentiation by influencing p38 MAPK signaling in lipopolysaccharide -induced human bone marrow mesenchymal stem cells

    International Nuclear Information System (INIS)

    Wang, Yuli; Wu, Hongxia; Shen, Ming; Ding, Siyang; Miao, Jing; Chen, Ning

    2017-01-01

    Periodontitis is a chronic inflammatory disease induced by bacterial pathogens, which not only affect connective tissue attachments but also cause alveolar bone loss. In this study, we investigated the anti-inflammatory effects of Human amnion-derived mesenchymal stem cells (HAMSCs) on human bone marrow mesenchymal stem cells (HBMSCs) under lipopolysaccharide (LPS)-induced inflammatory conditions. Proliferation levels were measured by flow cytometry and immunofluorescence staining of 5-ethynyl-2′-deoxyuridine (EdU). Osteoblastic differentiation and mineralization were investigated using chromogenic alkaline phosphatase activity (ALP) activity substrate assays, Alizarin red S staining, and RT-PCR analysis of HBMSCs osteogenic marker expression. Oxidative stress induced by LPS was investigated by assaying reactive oxygen species (ROS) level and superoxide dismutase (SOD) activity. Here, we demonstrated that HAMSCs increased the proliferation, osteoblastic differentiation, and SOD activity of LPS-induced HBMSCs, and down-regulated the ROS level. Moreover, our results suggested that the activation of p38 MAPK signal transduction pathway is essential for reversing the LPS-induced bone-destructive processes. SB203580, a selective inhibitor of p38 MAPK signaling, significantly suppressed the anti-inflammatory effects in HAMSCs. In conclusion, HAMSCs show a strong potential in treating inflammation-induced bone loss by influencing p38 MAPK signaling. - Highlights: • LPS inhibites osteogenic differentiation in HBMSCs via suppression of p38 MAPK signaling pathway. • HAMSCs promote LPS-induced HBMSCs osteogenic differentiation through p38 MAPK signaling pathway. • HAMSCs reverse LPS-induced oxidative stress in LPS-induced HBMSCs through p38 MAPK signaling pathway.

  13. Oesteosarcomagenic doses of radium (224Ra) and infectious endogenous retroviruses enhance proliferation and osteogenic differentiation of skeletal tissue dofferentiating in vitro

    International Nuclear Information System (INIS)

    Schmidt, J.; Heermeier, K.; Linzner, U.; Luz, A.; Silbermann, M.; Livne, E.; Erfle, V.

    1994-01-01

    Cartilage tissue from embryonic mice which undergoes osteogenic differentiation during in vitro cultivation was used to study the effect of osteosarcomagenic doses of α-irradiation and bone-tumor-inducing retroviruses on proliferation and phenotypic differentiation of skeletal cells in a defined tissue culture model. Irradiated mandibular condyles showed dose-dependent enhancement of cell proliferation at day 7 of the culture and increased osteogenic differentiation at day 14. Maximal effects were found with 7.4 Bq/ml of 224 Ra-labeled medium. Doses of 740 and 7400 Bq/ml of 224 Ra-labeled medium induced increasing cell death. Retrovirus infection enhanced osteogenic differentiation and extended the viability of irradiated cells. After transplantation none of the treated tissues developed tumors in syngeneic mice. (orig.)

  14. Human dental pulp stem cells derived from cryopreserved dental pulp tissues of vital extracted teeth with disease demonstrate hepatic-like differentiation.

    Science.gov (United States)

    Chen, Y K; Huang, Anderson H C; Chan, Anthony W S; Lin, L M

    2016-06-01

    Reviewing the literature, hepatic differentiation of human dental pulp stem cells (hDPSCs) from cryopreserved dental pulp tissues of vital extracted teeth with disease has not been studied. This study is aimed to evaluate the hypothesis that hDPSCs from cryopreserved dental pulp tissues of vital extracted teeth with disease could possess potential hepatic differentiation. Forty vital extracted teeth with disease recruited for hDPSCs isolation, stem cell characterization and hepatic differentiation were randomly and equally divided into group A (liquid nitrogen-stored dental pulp tissues) and group B (freshly derived dental pulp tissues). Samples of hDPSCs isolated from groups A and B but without hepatic growth factors formed negative controls. A well-differentiated hepatocellular carcinoma cell line was employed as a positive control. All the isolated hDPSCs from groups A and B showed hepatic-like differentiation with morphological change from a spindle-shaped to a polygonal shape and normal karyotype. Differentiated hDPSCs and the positive control expressed hepatic metabolic function genes and liver-specific genes. Glycogen storage of differentiated hDPSCs was noted from day 7 of differentiation-medium culture. Positive immunofluorescence staining of low-density lipoprotein and albumin was observed from day 14 of differentiation-medium culture; urea production in the medium was noted from week 6. No hepatic differentiation was observed for any of the samples of the negative controls. We not only demonstrated the feasibility of hepatic-like differentiation of hDPSCs from cryopreserved dental pulp tissues of vital extracted teeth with disease but also indicated that the differentiated cells possessed normal karyotype and were functionally close to normal hepatic-like cells. Copyright © 2013 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  15. Magnetohydrodynamic (MHD Jeffrey fluid over a stretching vertical surface in a porous medium

    Directory of Open Access Journals (Sweden)

    Kartini Ahmad

    2017-12-01

    Full Text Available This paper presents the study of steady two-dimensional mixed convection boundary layer flow and heat transfer of a Jeffrey fluid over a stretched sheet immersed in a porous medium in the presence of a transverse magnetic field. The governing partial differential equations are reduced to nonlinear ordinary differential equations with the aid of similarity transformation, which are then solved numerically using an implicit finite difference scheme. The effects of some of the embedded parameters, such as Deborah number β, magnetic parameter M, mixed convection parameter λ, porosity parameter γ and Prandtl number Pr, on the flow and heat transfer characteristics, are given in forms of tables and graphs.

  16. Determination of rivaroxaban in patient's plasma samples by anti-Xa chromogenic test associated to High Performance Liquid Chromatography tandem Mass Spectrometry (HPLC-MS/MS).

    Science.gov (United States)

    Derogis, Priscilla Bento Matos; Sanches, Livia Rentas; de Aranda, Valdir Fernandes; Colombini, Marjorie Paris; Mangueira, Cristóvão Luis Pitangueira; Katz, Marcelo; Faulhaber, Adriana Caschera Leme; Mendes, Claudio Ernesto Albers; Ferreira, Carlos Eduardo Dos Santos; França, Carolina Nunes; Guerra, João Carlos de Campos

    2017-01-01

    Rivaroxaban is an oral direct factor Xa inhibitor, therapeutically indicated in the treatment of thromboembolic diseases. As other new oral anticoagulants, routine monitoring of rivaroxaban is not necessary, but important in some clinical circumstances. In our study a high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method was validated to measure rivaroxaban plasmatic concentration. Our method used a simple sample preparation, protein precipitation, and a fast chromatographic run. It was developed a precise and accurate method, with a linear range from 2 to 500 ng/mL, and a lower limit of quantification of 4 pg on column. The new method was compared to a reference method (anti-factor Xa activity) and both presented a good correlation (r = 0.98, p plasma samples for rivaroxaban measurement by HPLC-MS/MS without interferences. The chromogenic and HPLC-MS/MS methods were highly correlated and should be used as clinical tools for drug monitoring. The method was applied successfully in a group of 49 real-life patients, which allowed an accurate determination of rivaroxaban in peak and trough levels.

  17. Micropropagation of Alstroemeria in liquid medium using slow release of medium components

    NARCIS (Netherlands)

    Klerk, de G.J.M.; Brugge, ter J.

    2010-01-01

    Alstroemeria rhizomes were micropropagated on semi-solid medium (AM) and in liquid medium (LM). In LM, growth was much enhanced (ca. 70%). Adequate gas exchange was crucial. This was obtained by agitation and in static medium by a sufficient large contact area of the explant and the gaseous

  18. Characterisation of urinary stones in the presence of iodinated contrast medium using dual-energy CT: a phantom study

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Jia; Qu, Mingliang; Duan, Xinhui; Takahashi, Naoki; Kawashima, Akira; Leng, Shuai; McCollough, Cynthia H. [Mayo Clinic, Department of Radiology, Rochester, MN (United States)

    2012-12-15

    To develop a dual-energy CT (DECT) method for differentiating uric acid (UA) from non-UA stones in the presence of iodinated contrast medium. Thirty UA and 45 non-UA stones were selected after infra-red spectroscopic analysis and independently placed in a 1.5-ml vial, which was filled first with saline and then with increasing concentrations of iodine. For each condition, tubes were put in a 35-cm water phantom and examined using a dual-source CT system at 100 and 140 kV. Virtual unenhanced images created from CT data sets of the stones in iodine-containing solutions provided position and volume information. This map was used to calculate a CT number ratio to differentiate stone type. A region-growing method was developed to improve the ability to differentiate between UA and non-UA stones with iodinated contrast medium. The sensitivity for detecting UA stones was 100 % for unenhanced images but fell to 18 % with 20 mgI/ml iodine solution and 0 % for higher concentrations. With region growing, the sensitivity for detecting UA stones was increased to 100 %, 82 %, 57 %, 50 % and 21 % for iodine solutions of 20, 40, 60, 80 and 100 mgI/ml. The region-growing method improves differentiation of UA from non-UA stones on contrast-enhanced DECT urograms. (orig.)

  19. Community effect triggers terminal differentiation of myogenic cells derived from muscle satellite cells by quenching Smad signaling

    Energy Technology Data Exchange (ETDEWEB)

    Yanagisawa, Michiko [Department of Regenerative Medicine, National Institute for Longevity Sciences, National Center for Geriatrics and Gerontology, 35 Gengo, Morioka, Oobu, Aichi 474-8522 (Japan); Aging Research, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, Aichi 466-8550 (Japan); Mukai, Atsushi; Shiomi, Kosuke [Department of Regenerative Medicine, National Institute for Longevity Sciences, National Center for Geriatrics and Gerontology, 35 Gengo, Morioka, Oobu, Aichi 474-8522 (Japan); Song, Si-Yong [Institute of Neuroscience, Faculty of Pharmaceutical Sciences at Kagawa, Tokushima Bunri University, 1314-1 Shido, Sanuki-shi, Kagawa 769-2193 (Japan); Hashimoto, Naohiro, E-mail: nao@ncgg.go.jp [Department of Regenerative Medicine, National Institute for Longevity Sciences, National Center for Geriatrics and Gerontology, 35 Gengo, Morioka, Oobu, Aichi 474-8522 (Japan)

    2011-01-15

    A high concentration of bone morphogenetic proteins (BMPs) stimulates myogenic progenitor cells to undergo heterotopic osteogenic differentiation. However, the physiological role of the Smad signaling pathway during terminal muscle differentiation has not been resolved. We report here that Smad1/5/8 was phosphorylated and activated in undifferentiated growing mouse myogenic progenitor Ric10 cells without exposure to any exogenous BMPs. The amount of phosphorylated Smad1/5/8 was severely reduced during precocious myogenic differentiation under the high cell density culture condition even in growth medium supplemented with a high concentration of serum. Inhibition of the Smad signaling pathway by dorsomorphin, an inhibitor of Smad activation, or noggin, a specific antagonist of BMP, induced precocious terminal differentiation of myogenic progenitor cells in a cell density-dependent fashion even in growth medium. In addition, Smad1/5/8 was transiently activated in proliferating myogenic progenitor cells during muscle regeneration in rats. The present results indicate that the Smad signaling pathway is involved in a critical switch between growth and differentiation of myogenic progenitor cells both in vitro and in vivo. Furthermore, precocious cell density-dependent myogenic differentiation suggests that a community effect triggers the terminal muscle differentiation of myogenic cells by quenching the Smad signaling.

  20. Community effect triggers terminal differentiation of myogenic cells derived from muscle satellite cells by quenching Smad signaling

    International Nuclear Information System (INIS)

    Yanagisawa, Michiko; Mukai, Atsushi; Shiomi, Kosuke; Song, Si-Yong; Hashimoto, Naohiro

    2011-01-01

    A high concentration of bone morphogenetic proteins (BMPs) stimulates myogenic progenitor cells to undergo heterotopic osteogenic differentiation. However, the physiological role of the Smad signaling pathway during terminal muscle differentiation has not been resolved. We report here that Smad1/5/8 was phosphorylated and activated in undifferentiated growing mouse myogenic progenitor Ric10 cells without exposure to any exogenous BMPs. The amount of phosphorylated Smad1/5/8 was severely reduced during precocious myogenic differentiation under the high cell density culture condition even in growth medium supplemented with a high concentration of serum. Inhibition of the Smad signaling pathway by dorsomorphin, an inhibitor of Smad activation, or noggin, a specific antagonist of BMP, induced precocious terminal differentiation of myogenic progenitor cells in a cell density-dependent fashion even in growth medium. In addition, Smad1/5/8 was transiently activated in proliferating myogenic progenitor cells during muscle regeneration in rats. The present results indicate that the Smad signaling pathway is involved in a critical switch between growth and differentiation of myogenic progenitor cells both in vitro and in vivo. Furthermore, precocious cell density-dependent myogenic differentiation suggests that a community effect triggers the terminal muscle differentiation of myogenic cells by quenching the Smad signaling.

  1. Human Mesenchymal Stem Cells Growth and Osteogenic Differentiation on Piezoelectric Poly(vinylidene fluoride Microsphere Substrates

    Directory of Open Access Journals (Sweden)

    R. Sobreiro-Almeida

    2017-11-01

    Full Text Available The aim of this work was to determine the influence of the biomaterial environment on human mesenchymal stem cell (hMSC fate when cultured in supports with varying topography. Poly(vinylidene fluoride (PVDF culture supports were prepared with structures ranging between 2D and 3D, based on PVDF films on which PVDF microspheres were deposited with varying surface density. Maintenance of multipotentiality when cultured in expansion medium was studied by flow cytometry monitoring the expression of characteristic hMSCs markers, and revealed that cells were losing their characteristic surface markers on these supports. Cell morphology was assessed by scanning electron microscopy (SEM. Alkaline phosphatase activity was also assessed after seven days of culture on expansion medium. On the other hand, osteoblastic differentiation was monitored while culturing in osteogenic medium after cells reached confluence. Osteocalcin immunocytochemistry and alizarin red assays were performed. We show that flow cytometry is a suitable technique for the study of the differentiation of hMSC seeded onto biomaterials, giving a quantitative reliable analysis of hMSC-associated markers. We also show that electrosprayed piezoelectric poly(vinylidene fluoride is a suitable support for tissue engineering purposes, as hMSCs can proliferate, be viable and undergo osteogenic differentiation when chemically stimulated.

  2. A chemically defined culture medium containing Rho kinase inhibitor Y-27632 for the fabrication of stratified squamous epithelial cell grafts

    International Nuclear Information System (INIS)

    Aslanova, Afag; Takagi, Ryo; Yamato, Masayuki; Okano, Teruo; Yamamoto, Masakazu

    2015-01-01

    With the development of a culture method for stratified squamous epithelial cells, tissue-engineered epithelial cell sheets have been successfully applied as clinical cell grafts. However, the implementation of these cell sheets without the use of any animal-derived materials is highly desirable. In this study, Rho-associated protein kinase inhibitor Y-27632 was used to develop a chemically defined culture medium for the fabrication of stratified epithelial cell grafts consisting of human epidermal and oral keratinocytes, and the proliferation activity, cell morphology, and gene expressions of the keratinocytes were analyzed. The results of a colorimetric assay indicated that Y-27632 significantly promoted the proliferation of the keratinocytes in culture media both with and without fetal bovine serum (FBS), although there were no indications of Y-27632 efficacy on cell morphology and stratification of the keratinocytes in culture medium without any animal-derived materials. The results of quantitative RT-PCR revealed that gene expressions correlated with cell adhesion, cell–cell junction, proliferation markers, and stem/progenitor markers in cultured keratinocytes were not strongly affected by the addition of Y-27632 to the culture medium. Moreover, gene expressions of differentiation markers in stratified keratinocytes cultured in medium without FBS were nearly identical to those of keratinocytes co-cultured with 3T3 feeder cells. Interestingly, the expressions of differentiation markers in cultured stratified keratinocytes were suppressed by FBS, whereas they were reconstructed by either co-culture of a 3T3 feeder layer or addition of Y-27632 into the culture medium containing FBS. These findings indicate that Y-27632 is a useful supplement for the development of a chemically defined culture medium for fabrication of stratified epithelial cell grafts for clinical applications for the purpose of developing the culture medium with a lower risk of pathogen

  3. A chemically defined culture medium containing Rho kinase inhibitor Y-27632 for the fabrication of stratified squamous epithelial cell grafts

    Energy Technology Data Exchange (ETDEWEB)

    Aslanova, Afag [Department of Surgery, Institute of Gastroenterology, Tokyo Women' s Medical University, 8-1 Kawada-cho, Shinjuku-ku, Tokyo 162-8666 (Japan); Institute of Advanced Biomedical Engineering and Science, Tokyo Women' s Medical University, TWIns, 8-1 Kawada-cho, Shinjuku-ku, Tokyo 162-8666 (Japan); Takagi, Ryo; Yamato, Masayuki; Okano, Teruo [Institute of Advanced Biomedical Engineering and Science, Tokyo Women' s Medical University, TWIns, 8-1 Kawada-cho, Shinjuku-ku, Tokyo 162-8666 (Japan); Yamamoto, Masakazu, E-mail: yamamoto.ige@twmu.ac.jp [Department of Surgery, Institute of Gastroenterology, Tokyo Women' s Medical University, 8-1 Kawada-cho, Shinjuku-ku, Tokyo 162-8666 (Japan)

    2015-05-01

    With the development of a culture method for stratified squamous epithelial cells, tissue-engineered epithelial cell sheets have been successfully applied as clinical cell grafts. However, the implementation of these cell sheets without the use of any animal-derived materials is highly desirable. In this study, Rho-associated protein kinase inhibitor Y-27632 was used to develop a chemically defined culture medium for the fabrication of stratified epithelial cell grafts consisting of human epidermal and oral keratinocytes, and the proliferation activity, cell morphology, and gene expressions of the keratinocytes were analyzed. The results of a colorimetric assay indicated that Y-27632 significantly promoted the proliferation of the keratinocytes in culture media both with and without fetal bovine serum (FBS), although there were no indications of Y-27632 efficacy on cell morphology and stratification of the keratinocytes in culture medium without any animal-derived materials. The results of quantitative RT-PCR revealed that gene expressions correlated with cell adhesion, cell–cell junction, proliferation markers, and stem/progenitor markers in cultured keratinocytes were not strongly affected by the addition of Y-27632 to the culture medium. Moreover, gene expressions of differentiation markers in stratified keratinocytes cultured in medium without FBS were nearly identical to those of keratinocytes co-cultured with 3T3 feeder cells. Interestingly, the expressions of differentiation markers in cultured stratified keratinocytes were suppressed by FBS, whereas they were reconstructed by either co-culture of a 3T3 feeder layer or addition of Y-27632 into the culture medium containing FBS. These findings indicate that Y-27632 is a useful supplement for the development of a chemically defined culture medium for fabrication of stratified epithelial cell grafts for clinical applications for the purpose of developing the culture medium with a lower risk of pathogen

  4. Simultaneous estimation of strength and position of a heat source in a participating medium using DE algorithm

    International Nuclear Information System (INIS)

    Parwani, Ajit K.; Talukdar, Prabal; Subbarao, P.M.V.

    2013-01-01

    An inverse heat transfer problem is discussed to estimate simultaneously the unknown position and timewise varying strength of a heat source by utilizing differential evolution approach. A two dimensional enclosure with isothermal and black boundaries containing non-scattering, absorbing and emitting gray medium is considered. Both radiation and conduction heat transfer are included. No prior information is used for the functional form of timewise varying strength of heat source. The finite volume method is used to solve the radiative transfer equation and the energy equation. In this work, instead of measured data, some temperature data required in the solution of the inverse problem are taken from the solution of the direct problem. The effect of measurement errors on the accuracy of estimation is examined by introducing errors in the temperature data of the direct problem. The prediction of source strength and its position by the differential evolution (DE) algorithm is found to be quite reasonable. -- Highlights: •Simultaneous estimation of strength and position of a heat source. •A conducting and radiatively participating medium is considered. •Implementation of differential evolution algorithm for such kind of problems. •Profiles with discontinuities can be estimated accurately. •No limitation in the determination of source strength at the final time

  5. Effects of fullerol on the osteogenic differentiation of adipose-derived mesenchymal stem cells

    Directory of Open Access Journals (Sweden)

    Yan-ting TANG

    2016-09-01

    Full Text Available Objective  To investigate the effect of fullerol on the osteogenic differentiation of rat adipose-derived mesenchymal stem cells (rADSCs based on hanging drop culture. Methods  rADSC spheres were formed by hanging drop culture for 3 days, then the spheres were dissociated to single cell by trypsin (called rADSC sphere-derived cells. The rADSC sphere-derived cells were cultured in 2-D adherent cell cultures for 24 hours, then the ordinary culture medium was replaced by osteogenic induction medium (osteogenic medium, OM. OM without fullerol served as control group (CM; OM with 1.0μmol/L fullerol was used as the experimental group (OM+Ful1.0μmol/L. Two groups were induced to differentiation for 14d and 21d, using two methods of alizarin red staining and quantitative real-time PCR (qPCR to detecte the ability of rADSC sphere-derived cells differentiating into osteoblasts. Results  rADSCs could form a uniform size microspheres structure through hanging drop culture for 3 days; rADSC sphere-derived cells were obtained by dissipation of trypsin. Compared with OM group, OM+Ful1.0μmol/L can make rADSC sphere-derived cells form more mineralized calcium nodules, and enhance the expression of the relevant osteogenic gene Runx2, OCN and ColI. Conclusion  Fullerol can significantly enhance the osteogenic differentiation of rADSCs based on hanging drop culture, and it is helpful to improve the efficiency of osteogenic induction of rADSCs. DOI: 10.11855/j.issn.0577-7402.2016.08.03

  6. The role of dual energy CT in differentiating between brain haemorrhage and contrast medium after mechanical revascularisation in acute ischaemic stroke

    International Nuclear Information System (INIS)

    Tijssen, M.P.M.; Stadler, A.A.R.; Zwam, W. van; Graaf, R. de; Postma, A.A.; Hofman, P.A.M.; Oostenbrugge, R.J. van; Klotz, E.; Wildberger, J.E.

    2014-01-01

    To assess the feasibility of dual energy computed tomography (DE-CT) in intra-arterially treated acute ischaemic stroke patients to discriminate between contrast extravasation and intracerebral haemorrhage. Thirty consecutive acute ischaemic stroke patients following intra-arterial treatment were examined with DE-CT. Simultaneous imaging at 80 kV and 140 kV was employed with calculation of mixed images. Virtual unenhanced non-contrast (VNC) images and iodine overlay maps (IOM) were calculated using a dedicated brain haemorrhage algorithm. Mixed images alone, as ''conventional CT'', and DE-CT interpretations were evaluated and compared with follow-up CT. Eight patients were excluded owing to a lack of follow-up or loss of data. Mixed images showed intracerebral hyperdense areas in 19/22 patients. Both haemorrhage and residual contrast material were present in 1/22. IOM suggested contrast extravasation in 18/22 patients; in 16/18 patients this was confirmed at follow-up. The positive predictive value (PPV) of mixed imaging alone was 25 %, with a negative predictive value (NPV) of 91 % and accuracy of 63 %. The PPV for detection of haemorrhage with DE-CT was 100 %, with an NPV of 89 % and accuracy improved to 89 %. Dual energy computed tomography improves accuracy and diagnostic confidence in early differentiation between intracranial haemorrhage and contrast medium extravasation in acute stroke patients following intra-arterial revascularisation. (orig.)

  7. The role of dual energy CT in differentiating between brain haemorrhage and contrast medium after mechanical revascularisation in acute ischaemic stroke

    Energy Technology Data Exchange (ETDEWEB)

    Tijssen, M.P.M.; Stadler, A.A.R.; Zwam, W. van; Graaf, R. de; Postma, A.A. [Maastricht University Medical Centre, Department of Radiology, P.O. Box 5800, Maastricht (Netherlands); Hofman, P.A.M. [Maastricht University Medical Centre, Department of Radiology, P.O. Box 5800, Maastricht (Netherlands); Maastricht University, MhENS School for Mental Health and Neuroscience, Maastricht (Netherlands); Oostenbrugge, R.J. van [Maastricht University Medical Centre, Department of Neurology, P.O. Box 5800, Maastricht (Netherlands); Maastricht University, CARIM School for Cardiovascular Diseases, P.O. Box 616, Maastricht (Netherlands); Klotz, E. [Siemens Healthcare Sector, Computed Tomography, Forchheim (Germany); Wildberger, J.E. [Maastricht University Medical Centre, Department of Radiology, P.O. Box 5800, Maastricht (Netherlands); Maastricht University, CARIM School for Cardiovascular Diseases, P.O. Box 616, Maastricht (Netherlands)

    2014-04-15

    To assess the feasibility of dual energy computed tomography (DE-CT) in intra-arterially treated acute ischaemic stroke patients to discriminate between contrast extravasation and intracerebral haemorrhage. Thirty consecutive acute ischaemic stroke patients following intra-arterial treatment were examined with DE-CT. Simultaneous imaging at 80 kV and 140 kV was employed with calculation of mixed images. Virtual unenhanced non-contrast (VNC) images and iodine overlay maps (IOM) were calculated using a dedicated brain haemorrhage algorithm. Mixed images alone, as ''conventional CT'', and DE-CT interpretations were evaluated and compared with follow-up CT. Eight patients were excluded owing to a lack of follow-up or loss of data. Mixed images showed intracerebral hyperdense areas in 19/22 patients. Both haemorrhage and residual contrast material were present in 1/22. IOM suggested contrast extravasation in 18/22 patients; in 16/18 patients this was confirmed at follow-up. The positive predictive value (PPV) of mixed imaging alone was 25 %, with a negative predictive value (NPV) of 91 % and accuracy of 63 %. The PPV for detection of haemorrhage with DE-CT was 100 %, with an NPV of 89 % and accuracy improved to 89 %. Dual energy computed tomography improves accuracy and diagnostic confidence in early differentiation between intracranial haemorrhage and contrast medium extravasation in acute stroke patients following intra-arterial revascularisation. (orig.)

  8. Human mesenchymal stem cells promote CD34+ hematopoietic stem cell proliferation with preserved red blood cell differentiation capacity.

    Science.gov (United States)

    Lau, Show Xuan; Leong, Yin Yee; Ng, Wai Hoe; Ng, Albert Wee Po; Ismail, Ida Shazrina; Yusoff, Narazah Mohd; Ramasamy, Rajesh; Tan, Jun Jie

    2017-06-01

    Studies showed that co-transplantation of mesenchymal stem cells (MSCs) and cord blood-derived CD34 + hematopoietic stem cells (HSCs) offered greater therapeutic effects but little is known regarding the effects of human Wharton's jelly derived MSCs on HSC expansion and red blood cell (RBC) generation in vitro. This study aimed to investigate the effects of MSCs on HSC expansion and differentiation. HSCs were co-cultured with MSCs or with 10% MSCs-derived conditioned medium, with HSCs cultured under standard medium served as a control. Cell expansion rates, number of mononuclear cell post-expansion and number of enucleated cells post-differentiation were evaluated. HSCs showed superior proliferation in the presence of MSC with mean expansion rate of 3.5 × 10 8  ± 1.8 × 10 7 after day 7 compared to the conditioned medium and the control group (8.9 × 10 7  ± 1.1 × 10 8 and 7.0 × 10 7  ± 3.3 × 10 6 respectively, P cell was greater compared to earlier passages, indicating successful RBC differentiation. Cord blood-derived CD34 + HSCs can be greatly expanded by co-culturing with MSCs without affecting the RBC differentiation capability, suggesting the importance of direct MSC-HSCs contact in HSC expansion and RBC differentiation. © 2017 International Federation for Cell Biology.

  9. Design, chemical synthesis and kinetic studies of trypsin chromogenic substrates based on the proteinase binding loop of Cucurbita maxima trypsin inhibitor (CMTI-III).

    Science.gov (United States)

    Lesner, A; Brzozowski, K; Kupryszewski, G; Rolka, K

    2000-03-05

    A series of trypsin chromogenic substrates with formula: Y-Ala-X-Abu-Pro-Lys-pNA, where X = Gly, Ala, Abu, Val, Leu, Phe, Ser, Glu and Y = Ac, H; pNA = p-nitroanilide was synthesized. The Cucurbita maxima trypsin inhibitor CMTI-III molecule was used as a vehicle to design the trypsin substrates. To evaluate the influence of position P(4) on the substrate-enzyme interaction, kinetic parameters of newly synthesized substrates with bovine beta-trypsin were determined. The increasing hydrophobicity of the amino acid residue (Gly, Ala, Abu, Val) introduced in position P(4) significantly enhanced the substrate specificity (k(cat)/K(m)) which was over 8 times higher for the last residue than that for the first one. The introduction of residues with more hydrophilic side chain (Glu, Ser) in this position reduced the value of this parameter. These results correspond well with those obtained using molecular dynamics of bovine beta-trypsin with monosubstituted CMTI-I analogues, indicating that in both trypsin substrate and inhibitor position 4 plays an important role in the interaction with the enzyme. Copyright 2000 Academic Press.

  10. Structure of Small and Medium-Sized Business: Results of Total Statistic Observations in Russia

    Directory of Open Access Journals (Sweden)

    Iuliia S. Pinkovetskaia

    2018-03-01

    Full Text Available The aim of the research is estimation of regularities and tendencies, characteristic for modern sectoral structure of small and mediumsized business in Russia. The subject of the research is a set of processes of structural changes on the types of economic activities of such enterprises, as well as the differentiation of the number of employees in enterprises. The research methodology included consideration of aggregates of subjects of small and medium-sized business, formed according to sectoral and territorial features. As the initial data used the official statistical information, which was obtain in the course of total observation of the activities of small and medium-sized businesses in 2010 and 2015. The study was conducted on indicators characterizing the full range of legal entities and individual entrepreneurs in the country. The materiality of structural changes was carried out on the basis of the Ryabtsev index. Modeling the differentiation of the values of the number of employees per enterprise was based on the development of density normal distribution functions. According to the hypothesis it is assumed that the differentiation of the number of employees working in enterprises depend on six main types of economic activity and on the subjects of Russia. Based on the results of the study was proved that there are no significant structural changes for the period from 2010 to 2015, both in terms of the number of enterprises and the number of their employees. Based on the results of the simulation, the average values of the number of employees for the six main types of activity were established, as well as the intervals for changing these indicators for the aggregates of small and medium-sized enterprises located in the majority of the country's subjects. The results of research can be used in the performance of scientific works related to the justification of the expected number and number of employees of enterprises, the formation of

  11. Agreement between chromogenic in situ hybridisation (CISH) and FISH in the determination of HER2 status in breast cancer.

    Science.gov (United States)

    Arnould, L; Denoux, Y; MacGrogan, G; Penault-Llorca, F; Fiche, M; Treilleux, I; Mathieu, M C; Vincent-Salomon, A; Vilain, M O; Couturier, J

    2003-05-19

    Determination of the HER2/neu (HER2) status in breast carcinoma has become necessary for the selection of breast cancer patients for trastuzumab therapy. Amplification of the gene analysed by fluorescence in situ hybridisation (FISH) or overexpression of the protein determined by immunohistochemistry (IHC) are the two major methods to establish this status. A strong correlation has been previously demonstrated between these two methods. However, FISH is not always feasible in routine practice and weakly positive IHC tumours (2+) do not always correspond to a gene amplification. Our study was performed in order to evaluate the contribution of chromogenic in situ hybridisation (CISH), which enables detection of the gene copies through an immunoperoxidase reaction. CISH was performed in 79 breast carcinomas for which the HER2 status was previously determined by IHC and FISH. The results of IHC, FISH and CISH were compared for each tumour. CISH procedures were successful in 95% of our cases. Whatever the IHC results, we found a very good concordance (96%) between CISH and FISH. Our study confirms that CISH may be an alternative to FISH for the determination of the gene amplification status in 2+ tumours. Our results allow us to think that, in many laboratories, CISH may also be an excellent method to calibrate the IHC procedures or, as a quality control test, to check regularly that the IHC signal is in agreement with the gene status.

  12. Three-dimensional radiative transfer in an isotropically scattering, plane-parallel medium: generalized X- and Y-functions

    International Nuclear Information System (INIS)

    Mueller, D.W.; Crosbie, A.L.

    2005-01-01

    The topic of this work is the generalized X- and Y-functions of multidimensional radiative transfer. The physical problem considered is spatially varying, collimated radiation incident on the upper boundary of an isotropically scattering, plane-parallel medium. An integral transform is used to reduce the three-dimensional transport equation to a one-dimensional form, and a modified Ambarzumian's method is used to derive coupled, integro-differential equations for the source functions at the boundaries of the medium. The resulting equations are said to be in double-integral form because the integration is over both angular variables. Numerical results are presented to illustrate the computational characteristics of the formulation

  13. Determining the correlation of Epstein-Barr virus with diffuse large B-cell lymphoma by chromogenic in situ hybridization

    Directory of Open Access Journals (Sweden)

    Kosari F

    2012-09-01

    Full Text Available Background: Diffuse large B-cell lymphoma (DLBCL is the most common type of lymphoma. There are various types of DLBCL including immunoblastic and centroblastic. Epstein-Barr virus (EBV is a member of Herpes virus family found in all human populations inducing different lymphoproliferative disorders. The role of EBV in the development of DLBCL is known. Multiple laboratory methods are available for detecting EBV. This study was conducted to determine the correlation of EBV with DLBCL in samples referred to pathology ward in Shariati and Sina Hospitals by chromogenic in situ hybridization (CISH method.Methods: In this case/control study, pathological specimens of 50 patients with DLBCL as well as 50 reactive lymph nodes and tonsils (control group were collected from archives of Shariati and Sina Hospitals and were evaluated for EBV encoded RNA (EBER expression based on CISH method. A peptide nucleic acid (PNA EBV probe (Dakocytomatin was used while all the processes were done in RNAase-free conditions using RNAase-free water, sterile gloves and samplers. Results: Out of fifty specimens in the case group, eight were positive for EBER in comparison with two in the control group (P=0.046. No statistically significant difference was observed between intranodal or extranodal samples (P=0.736 or between males and females (P=0.0746.Conclusion: Our study showed that EBV positivity for EBER in patient with DLBCL could be determined more effectively by CISH method than immunohistochemistry (IHC. Comparative analysis between CISH, PCR and IHC methods is recommended.

  14. Chromogenic in situ hybridisation (CISH) is a powerful method to detect ALK-positive non-small cell lung carcinomas.

    Science.gov (United States)

    Wagner, F; Streubel, A; Roth, A; Stephan-Falkenau, S; Mairinger, T

    2014-05-01

    We assessed the potential of a chromogenic in situ hybridisation (CISH) assay in comparison with quantitative reverse transcription (RT)-PCR (qPCR) to detect anaplastic lymphoma kinase (ALK) break apart-positive lung carcinomas. Dual-colour CISH using a break apart probe for the ALK gene on 2p23 was performed with 181 formalin-fixed, paraffin-embedded tissue and agar block sections from 175 cases of non-small cell lung carcinomas (NSCLC). Stained slides were analysed with a standard bright-field microscope at 1000× magnification by counting signals from 60 non-overlapping nuclei from three different tumour areas. Samples with ≥15% of positive nuclei were judged as ALK break apart-positive. All samples were simultaneously analysed by qPCR for EML4-ALK to validate CISH results, and positive samples were subject to Sanger sequencing. CISH was successful with 173 of 181 hybridised samples (96%), and seven ALK break apart-positive cases were detected. CISH signals were specific and distinct for both colours. All positive cases were confirmed by qPCR and Sanger sequencing, and concordance between CISH and qPCR was 100%. Nearly all samples (9/10) which failed by qPCR were accessible to CISH analysis. CISH is a very reliable, convenient and inexpensive method to detect ALK-positive NSCLC. CISH success rate is comparably high as with qPCR, and it detects all ALK break apart events in a single assay. It is of special value when RNA quality is poor, or when small biopsies with a very limited amount of tumour cells have to be analysed.

  15. OP9 bone marrow stroma cells differentiate into megakaryocytes and platelets.

    Directory of Open Access Journals (Sweden)

    Yumiko Matsubara

    Full Text Available Platelets are essential for hemostatic plug formation and thrombosis. The mechanisms of megakaryocyte (MK differentiation and subsequent platelet production from stem cells remain only partially understood. The manufacture of megakaryocytes (MKs and platelets from cell sources including hematopoietic stem cells and pluripotent stem cells have been highlighted for studying the platelet production mechanisms as well as for the development of new strategies for platelet transfusion. The mouse bone marrow stroma cell line OP9 has been widely used as feeder cells for the differentiation of stem cells into MK lineages. OP9 cells are reported to be pre-adipocytes. We previously reported that 3T3-L1 pre-adipocytes differentiated into MKs and platelets. In the present study, we examined whether OP9 cells differentiate into MKs and platelets using MK lineage induction (MKLI medium previously established to generate MKs and platelets from hematopoietic stem cells, embryonic stem cells, and pre-adipocytes. OP9 cells cultured in MKLI medium had megakaryocytic features, i.e., positivity for surface markers CD41 and CD42b, polyploidy, and distinct morphology. The OP9-derived platelets had functional characteristics, providing the first evidence for the differentiation of OP9 cells into MKs and platelets. We then analyzed gene expressions of critical factors that regulate megakaryopoiesis and thrombopoiesis. The gene expressions of p45NF-E2, FOG, Fli1, GATA2, RUNX1, thrombopoietin, and c-mpl were observed during the MK differentiation. Among the observed transcription factors of MK lineages, p45NF-E2 expression was increased during differentiation. We further studied MK and platelet generation using p45NF-E2-overexpressing OP9 cells. OP9 cells transfected with p45NF-E2 had enhanced production of MKs and platelets. Our findings revealed that OP9 cells differentiated into MKs and platelets in vitro. OP9 cells have critical factors for megakaryopoiesis and

  16. Longevity of U cells of differentiated yeast colonies grown on respiratory medium depends on active glycolysis.

    Science.gov (United States)

    Čáp, Michal; Váchová, Libuše; Palková, Zdena

    2015-01-01

    Colonies of Saccharomyces cerevisiae laboratory strains pass through specific developmental phases when growing on solid respiratory medium. During entry into the so-called alkali phase, in which ammonia signaling is initiated, 2 prominent cell types are formed within the colonies: U cells in upper colony regions, which have a longevity phenotype and activate the expression of a large number of metabolic genes, and L cells in lower regions, which die more quickly and exhibit a starvation phenotype. Here, we performed a detailed analysis of the activities of enzymes of central carbon metabolism in lysates of both cell types and determined several fermentation end products, showing that previously reported expression differences are reflected in the different enzymatic capabilities of each cell type. Hence, U cells, despite being grown on respiratory medium, behave as fermenting cells, whereas L cells rely on respiratory metabolism and possess active gluconeogenesis. Using a spectrum of different inhibitors, we showed that glycolysis is essential for the formation, and particularly, the survival of U cells. We also showed that β-1,3-glucans that are released from the cell walls of L cells are the most likely source of carbohydrates for U cells.

  17. Analysis of Human Papilloma Virus 16/18 DNA and its Correlation with p16 Expression in Oral Cavity Squamous Cell Carcinoma in North-Eastern India: A Chromogenic in-situ Hybridization Based Study

    Science.gov (United States)

    Jitani, Ankit Kumar; Mishra, Jaya; Shunyu, N. Brian; Khonglah, Yookarin; Medhi, Jayanta

    2015-01-01

    Background The incidence of head and neck squamous cell carcinoma (HNSCC) is quite high in North Eastern India. Apart from the traditional risk factors like tobacco and alcohol consumption, human papilloma virus (HPV) is now considered an established causative agent. These HPV related tumour have a clinico-pathological profile that is quite divergent from conventional non-HPV related tumours. Association of HPV in oral cancers has not been explored in north-east India. Materials and Methods Thirty-one patients with oral cavity squamous cell carcinoma (OSCC) on treatment from October 2010 to January 2013 were included in the study. Patients who received neo-adjuvant chemotherapy were excluded. HPV 16/18 DNA was evaluated using Chromogenic in-situ Hybridization (CISH). Presence of nuclear signals was taken as positive HPV expression. p16 was evaluated using immunohistochemistry and was considered positive if ≥ 80% of the tumour cells showed strong and diffuse nuclear/cytoplasmic immunostaining. The results were analysed using Fisher exact test and confidence interval was calculated where required. Results The study group age ranged from 30 to 80 years (median age- 54.2 years). The most common site was gum, with well differentiated squamous cell carcinoma being the most common histology. HPV 16/18 DNA was positive in 29% (95% CI: 13.03% - 44.97%) cases and had a clear tendency towards statistical significance with non-smoker cases (p=0.05), lymph node metastasis (p=0.05) and a significant correlation with p16 overexpression (p=0.04). There was no significant correlation with other clinico-pathological parameters. Conclusion HPV 16/18 is associated with OSCC, commonly seen among non-smokers and may be related to nodal metastasis. So, HPV may be used as a prognostic factor in OSCC and p16 may be considered as a surrogate marker for HPV. PMID:26435951

  18. The hedgehog-signaling pathway is repressed during the osteogenic differentiation of dental follicle cells

    DEFF Research Database (Denmark)

    Morsczeck, Christian; Reck, A; Beck, H C

    2017-01-01

    of repressors of the hedgehog-signaling pathway such as Patched 1 (PTCH1), Suppressor of Fused (SUFU), and Parathyroid Hormone-Related Peptide (PTHrP). Previous studies suggested that hedgehog proteins induce the osteogenic differentiation of mesenchymal stem cells via a paracrine pathway. Indian hedgehog (IHH......) induced the expression of the osteogenic transcription factor RUNX2. However, a supplementation of the BMP2-based osteogenic differentiation medium with IHH did not induce the expression of RUNX2. Moreover, IHH inhibited slightly the ALP activity and the mineralization of osteogenic-differentiated DFCs...

  19. Matrix-directed differentiation of human adipose-derived mesenchymal stem cells to dermal-like fibroblasts that produce extracellular matrix.

    Science.gov (United States)

    Sivan, Unnikrishnan; Jayakumar, K; Krishnan, Lissy K

    2016-10-01

    Commercially available skin substitutes lack essential non-immune cells for adequate tissue regeneration of non-healing wounds. A tissue-engineered, patient-specific, dermal substitute could be an attractive option for regenerating chronic wounds, for which adipose-derived mesenchymal stem cells (ADMSCs) could become an autologous source. However, ADMSCs are multipotent in nature and may differentiate into adipocytes, osteocytes and chondrocytes in vitro, and may develop into undesirable tissues upon transplantation. Therefore, ADMSCs committed to the fibroblast lineage could be a better option for in vitro or in vivo skin tissue engineering. The objective of this study was to standardize in vitro culture conditions for ADMSCs differentiation into dermal-like fibroblasts which can synthesize extracellular matrix (ECM) proteins. Biomimetic matrix composite, deposited on tissue culture polystyrene (TCPS), and differentiation medium (DM), supplemented with fibroblast-conditioned medium and growth factors, were used as a fibroblast-specific niche (FSN) for cell culture. For controls, ADMSCs were cultured on bare TCPS with either DM or basal medium (BM). Culture of ADMSCs on FSN upregulated the expression of differentiation markers such as fibroblast-specific protein-1 (FSP-1) and a panel of ECM molecules specific to the dermis, such as fibrillin-1, collagen I, collagen IV and elastin. Immunostaining showed the deposition of dermal-specific ECM, which was significantly higher in FSN compared to control. Fibroblasts derived from ADMSCs can synthesize elastin, which is an added advantage for successful skin tissue engineering as compared to fibroblasts from skin biopsy. To obtain rapid differentiation of ADMSCs to dermal-like fibroblasts for regenerative medicine, a matrix-directed differentiation strategy may be employed. Copyright © 2014 John Wiley & Sons, Ltd. Copyright © 2014 John Wiley & Sons, Ltd.

  20. An Optical Biosensing Strategy Based on Selective Light Absorption and Wavelength Filtering from Chromogenic Reaction

    Directory of Open Access Journals (Sweden)

    Hyeong Jin Chun

    2018-03-01

    Full Text Available To overcome the time and space constraints in disease diagnosis via the biosensing approach, we developed a new signal-transducing strategy that can be applied to colorimetric optical biosensors. Our study is focused on implementation of a signal transduction technology that can directly translate the color intensity signals—that require complicated optical equipment for the analysis—into signals that can be easily counted with the naked eye. Based on the selective light absorption and wavelength-filtering principles, our new optical signaling transducer was built from a common computer monitor and a smartphone. In this signal transducer, the liquid crystal display (LCD panel of the computer monitor served as a light source and a signal guide generator. In addition, the smartphone was used as an optical receiver and signal display. As a biorecognition layer, a transparent and soft material-based biosensing channel was employed generating blue output via a target-specific bienzymatic chromogenic reaction. Using graphics editor software, we displayed the optical signal guide patterns containing multiple polygons (a triangle, circle, pentagon, heptagon, and 3/4 circle, each associated with a specified color ratio on the LCD monitor panel. During observation of signal guide patterns displayed on the LCD monitor panel using a smartphone camera via the target analyte-loaded biosensing channel as a color-filtering layer, the number of observed polygons changed according to the concentration of the target analyte via the spectral correlation between absorbance changes in a solution of the biosensing channel and color emission properties of each type of polygon. By simple counting of the changes in the number of polygons registered by the smartphone camera, we could efficiently measure the concentration of a target analyte in a sample without complicated and expensive optical instruments. In a demonstration test on glucose as a model analyte, we

  1. An Optical Biosensing Strategy Based on Selective Light Absorption and Wavelength Filtering from Chromogenic Reaction.

    Science.gov (United States)

    Chun, Hyeong Jin; Han, Yong Duk; Park, Yoo Min; Kim, Ka Ram; Lee, Seok Jae; Yoon, Hyun C

    2018-03-06

    To overcome the time and space constraints in disease diagnosis via the biosensing approach, we developed a new signal-transducing strategy that can be applied to colorimetric optical biosensors. Our study is focused on implementation of a signal transduction technology that can directly translate the color intensity signals-that require complicated optical equipment for the analysis-into signals that can be easily counted with the naked eye. Based on the selective light absorption and wavelength-filtering principles, our new optical signaling transducer was built from a common computer monitor and a smartphone. In this signal transducer, the liquid crystal display (LCD) panel of the computer monitor served as a light source and a signal guide generator. In addition, the smartphone was used as an optical receiver and signal display. As a biorecognition layer, a transparent and soft material-based biosensing channel was employed generating blue output via a target-specific bienzymatic chromogenic reaction. Using graphics editor software, we displayed the optical signal guide patterns containing multiple polygons (a triangle, circle, pentagon, heptagon, and 3/4 circle, each associated with a specified color ratio) on the LCD monitor panel. During observation of signal guide patterns displayed on the LCD monitor panel using a smartphone camera via the target analyte-loaded biosensing channel as a color-filtering layer, the number of observed polygons changed according to the concentration of the target analyte via the spectral correlation between absorbance changes in a solution of the biosensing channel and color emission properties of each type of polygon. By simple counting of the changes in the number of polygons registered by the smartphone camera, we could efficiently measure the concentration of a target analyte in a sample without complicated and expensive optical instruments. In a demonstration test on glucose as a model analyte, we could easily measure the

  2. Role of Relationship Marketing in Small and Medium-Sized Entreprises

    Directory of Open Access Journals (Sweden)

    Ružica Butigan

    2011-06-01

    Full Text Available Along with marketing, small and medium enterprises (SMEs are commonly associated in literature with relationship marketing which results in marketing networks. This paper examines the specific characteristics that differentiate large companies from small and medium-sized enterprises, and the reasons that prevent SMEs from engaging in traditional marketing within the scope of marketing mix. The paper also shows that the key characteristic which distinguishes small from large companies is a prominent role of owner/managers. The owner/manager affects marketing of SMEs by creating his or her own personal networks with stakeholders in the environment, and considers it an important strategic focus and a factor that increases the performance of marketing decision-making. After a thorough review of secondary data sources this paper examines relationship marketing as a marketing framework for SMEs and, finally, proves that relationship marketing is more appropriate for SMEs than are traditional marketing concepts, that relationship marketing and network marketing create an important framework for SMEs and that there is a link between relationship marketing, personal networks and SMEs.

  3. Elastic light scattering for clinical pathogens identification: application to early screening of Staphylococcus aureus on specific medium

    Science.gov (United States)

    Schultz, E.; Genuer, V.; Marcoux, P.; Gal, O.; Belafdil, C.; Decq, D.; Maurin, Max; Morales, S.

    2018-02-01

    Elastic Light Scattering (ELS) is an innovative technique to identify bacterial pathogens directly on culture plates. Compelling results have already been reported for agri-food applications. Here, we have developed ELS for clinical diagnosis, starting with Staphylococcus aureus early screening. Our goal is to bring a result (positive/negative) after only 6 h of growth to fight surgical-site infections. The method starts with the acquisition of the scattering pattern arising from the interaction between a laser beam and a single bacterial colony growing on a culture medium. Then, the resulting image, considered as the bacterial species signature, is analyzed using statistical learning techniques. We present a custom optical setup able to target bacterial colonies with various sizes (30-500 microns). This system was used to collect a reference dataset of 38 strains of S. aureus and other Staphyloccocus species (5459 images) on ChromIDSAID/ MRSA bi-plates. A validation set from 20 patients has then been acquired and clinically-validated according to chromogenic enzymatic tests. The best correct-identification rate between S. aureus and S. non-aureus (94.7%) has been obtained using a support vector machine classifier trained on a combination of Fourier-Bessel moments and Local- Binary-Patterns extracted features. This statistical model applied to the validation set provided a sensitivity and a specificity of 90.0% and 56.9%, or alternatively, a positive predictive value of 47% and a negative predictive value of 93%. From a clinical point of view, the results head in the right direction and pave the way toward the WHO's requirements for rapid, low-cost, and automated diagnosis tools.

  4. Deduction of the in-medium gluon distribution from photon-gluon fusion processes in peripheral ultrarelativistic heavy-ion collisions

    International Nuclear Information System (INIS)

    Greiner, M.; Hofmann, C.; Schaefer, A.; Soff, G.

    1994-08-01

    The photon-gluon fusion process into a pair of heavy quarks is studied for peripheral Pb+Pb collisions at LHC energies. The double differential cross section with respect to the invariant mass and the rapidity of the produced quark pair at zero rapidity is directly proportional to the gluon distribution in the nuclear medium. Differential cross sections for the b-quark production lie well in the μbarn/GeV regime which will allow the deduction of the gluon distribution at low x. Rapidity cuts for the actual detectors are also considered. (orig.)

  5. OPTIMIZATION OF HORMONE COMPOSITION OF NUTRIENT MEDIUM FOR IN VITRO EFFICIENT REGENERATION OF BREAD WHEAT

    Directory of Open Access Journals (Sweden)

    E. D. Nikitina

    2016-08-01

    Full Text Available Optimal values of phytohormones in the differential nutrient medium providing the efficient realization of morphogenetic potencies of four spring bread wheat varieties (Skala, Spectr, Zarnitsa and Zhnitsa from immature embryo cultures have been determined. For callus induction explants 1.5 – 1.7 mmin size were used, which were subsequently passed to the medium by Linsmaier&Skoog possessing 0.8 % of agar, 3 % of sucrose and 2.0 mg l-1 dichlorophenoxyacetic acid (2,4-D. Cell cultures were incubated in darkness at the temperature 26±1 °С. 30 – 35 days after in accordance with the scheme of complete factorial experiment of 32 type calli were passed to differential medium supplemented with 2,4-D at levels 0.5; 2.5; 4.0 mg l-1 and with kinetin (6-furfurylaminopurine at levels 0.5; 2.25 and 4.0 mg l-1. Number of replications for each of 9 variants was four. As a result, 20 mathematic models (4 varieties × 5 stages of regeneration designed as polynomial quadric equation were obtained. On the ground of the analysis of models it was established that optimal values for factors are not equal both for cultures of genotypes analyzed and for different regeneration stages. For callus tissues of Skala and Spectr an optimal value of kinetin for all regeneration stages was 0.5 mg l-1 except for the frequency of morphogenesis. Optimal values of 2,4-D for the same varieties were within 2.3 – 3.2 mg l-1. For cell cultures of Zarnitsa and Zhnitsa recommended concentration intervals made up 1.3 – 2.2 mg l-1 on kinetin except for the frequency of rhizogenesis, and 1.9 – 2.7 on 2,4-D. The level of exogenous phytohormones necessary for stem differentiation was lower than the one for root formation. The dependence of morphogenesis results on the hormonal status of the explant has been discussed.

  6. Vascular smooth muscle cell differentiation to an osteogenic phenotype involves matrix metalloproteinase-2 modulation by homocysteine.

    Science.gov (United States)

    Liu, Tingjiao; Lin, Jinghan; Ju, Ting; Chu, Lei; Zhang, Liming

    2015-08-01

    Arterial calcification is common in vascular diseases and involves conversion of vascular smooth muscle cells (VSMCs) to an osteoblast phenotype. Clinical studies suggest that the development of atherosclerosis can be promoted by homocysteine (HCY), but the mechanisms remain unclear. Here, we determined whether increases in HCY levels lead to an increase in VSMC calcification and differentiation, and examined the role of an extracellular matrix remodeler, matrix metalloproteinase-2 (MMP-2). Rat VSMCs were exposed to calcification medium in the absence or presence of HCY (10, 100 or 200 μmol/L) or an MMP-2 inhibitor (10(-6) or 10(-5) mol/L). MTT assays were performed to determine the cytotoxicity of the MMP-2 inhibitor in calcification medium containing 200 μmol/L HCY. Calcification was assessed by measurements of calcium deposition and alkaline phosphatase (ALP) activity as well as von Kossa staining. Expression of osteocalcin, bone morphogenetic protein (BMP)-2, and osteopontin, and MMP-2 was determined by immunoblotting. Calcification medium induced osteogenic differentiation of VSMCs. HCY promoted calcification, increased osteocalcin and BMP-2 expression, and decreased expression of osteopontin. MMP-2 expression was increased by HCY in a dose-dependent manner in VSMCs exposed to both control and calcification medium. The MMP-2 inhibitor decreased the calcium content and ALP activity, and attenuated the osteoblastic phenotype of VSMCs. Vascular calcification and osteogenic differentiation of VSMCs were positively regulated by HCY through increased/restored MMP-2 expression, increased expression of calcification proteins, and decreased anti-calcification protein levels. In summary, MMP-2 inhibition may be a protective strategy against VSMC calcification.

  7. Regenerative medicine for Parkinson's disease using differentiated nerve cells derived from human buccal fat pad stem cells.

    Science.gov (United States)

    Takahashi, Haruka; Ishikawa, Hiroshi; Tanaka, Akira

    2017-04-01

    The purpose of this study was to evaluate the utility of human adipose stem cells derived from the buccal fat pad (hBFP-ASCs) for nerve regeneration. Parkinson's disease (PD) is a neurodegenerative disorder characterized by progressive death of dopaminergic neurons. PD is a candidate disease for cell replacement therapy because it has no fundamental therapeutic methods. We examined the properties of neural-related cells induced from hBFP-ASCs as a cell source for PD treatment. hBFP-ASCs were cultured in neurogenic differentiation medium for about 2 weeks. After the morphology of hBFP-ASCs changed to neural-like cells, the medium was replaced with neural maintenance medium. Cells differentiated from hBFP-ASCs showed neuron-like structures and expressed neuron markers (β3-tubulin, neurofilament 200, and microtubule-associated protein 2), an astrocyte marker (glial fibrillary acidic protein), or dopaminergic neuron-related marker (tyrosine hydroxylase). Induced neural cells were transplanted into a 6-hydroxydopamine (6-OHDA)-lesioned rat hemi-parkinsonian model. At 4 weeks after transplantation, 6-OHDA-lesioned rats were subjected to apomorphine-induced rotation analysis. The transplanted cells survived in the brain of rats as dopaminergic neural cells. No tumor formation was found after cell transplantation. We demonstrated differentiation of hBFP-ASCs into neural cells, and that transplantation of these neural cells improved the symptoms of model rats. Our results suggest that neurons differentiated from hBFP-ASCs would be applicable to cell replacement therapy of PD.

  8. A differentiated plane wave: its passage through a slab

    International Nuclear Information System (INIS)

    Hannay, J H; Nye, J F

    2013-01-01

    Differentiating a monochromatic uniform plane electromagnetic wavefield with respect to its direction produces, from a field that is completely lacking in localized specific features, one that contains a straight vortex-like line, a ‘C-line’ of defined circular polarization. There is also a second separate C-line of opposite handedness; indeed, in a sense, a straight line of every polarization is realized. Because of its primitive construction it is analytically simple to study the passage of a differentiated wave obliquely through a plane interface into a medium of different refractive index, to trace its C-line. This was done in an earlier paper. Here we extend the method to passage through a parallel-sided transparent slab. There are multiple reflections within the slab, as in a Fabry–Pérot interferometer. The exiting wave, as a single differentiated plane wave, has a straight oblique C-line. Inside the slab, and in front of it, there is wave interference. The result is a coiled, helix-like, C-line in front of the slab and another inside it. The two coils wrap around separate hyperboloids of one sheet, like cooling towers. The emerging straight C-line is shifted (with respect to a C-line in a notional undisturbed incident plane wave) both in the plane of incidence and transversely to it, and the second C-line behaves similarly. The analysis is exact and could be extended in a straightforward way to a general stratified medium. (paper)

  9. Pericytes Stimulate Oligodendrocyte Progenitor Cell Differentiation during CNS Remyelination

    Directory of Open Access Journals (Sweden)

    Alerie Guzman De La Fuente

    2017-08-01

    Full Text Available The role of the neurovascular niche in CNS myelin regeneration is incompletely understood. Here, we show that, upon demyelination, CNS-resident pericytes (PCs proliferate, and parenchymal non-vessel-associated PC-like cells (PLCs rapidly develop. During remyelination, mature oligodendrocytes were found in close proximity to PCs. In Pdgfbret/ret mice, which have reduced PC numbers, oligodendrocyte progenitor cell (OPC differentiation was delayed, although remyelination proceeded to completion. PC-conditioned medium accelerated and enhanced OPC differentiation in vitro and increased the rate of remyelination in an ex vivo cerebellar slice model of demyelination. We identified Lama2 as a PC-derived factor that promotes OPC differentiation. Thus, the functional role of PCs is not restricted to vascular homeostasis but includes the modulation of adult CNS progenitor cells involved in regeneration.

  10. Numerical Simulation of Transient Free Convection Flow and Heat Transfer in a Porous Medium

    Directory of Open Access Journals (Sweden)

    Rajesh Sharma

    2013-01-01

    Full Text Available The coupled momentum and heat transfer in unsteady, incompressible flow along a semi-infinite vertical porous moving plate adjacent to an isotropic porous medium with viscous dissipation effect are investigated. The Darcy-Forchheimer nonlinear drag force model which includes the effects of inertia drag forces is employed. The governing differential equations of the problem are transformed into a system of nondimensional differential equations which are solved numerically by the finite element method (FEM. The non-dimensional velocity and temperature profiles are presented for the influence of Darcy number, Forchheimer number, Grashof number, Eckert number, Prandtl number, plate velocity, and time. The Nusselt number is also evaluated and compared with finite difference method (FDM, which shows excellent agreement.

  11. Tubular and endothelial chimerism in renal allografts using fluorescence and chromogenic in situ hybridization (FISH, CISH) technology.

    Science.gov (United States)

    Varga, Zsuzsanna; Gaspert, Ariana; Behnke, Silvia; von Teichman, Adriana; Fritzsche, Florian; Fehr, Thomas

    2012-04-01

    The role of endothelial and tubular chimerism in renal allograft adaptation and rejection varies in different studies. We addressed the correlation between different clinico-pathological settings and sex-chromosomal endothelial and/or tubular chimerism in renal allografts. We examined the presence or absence of the X and Y chromosomes by fluorescence and chromogenic in situ hybridization (FISH, CISH) methodology on paraffin embedded kidney biopsies in 16 gender mismatched renal transplants (1 to 12 years post-transplantation). Twelve patients were male, four female. Four groups were selected: (i) Vascular calcineurin inhibitor toxicity without rejection; (ii) T-cell mediated vascular rejection; (iii) antibody mediated rejection; and (iv) C4d-positivity in AB0-incompatible transplants with or without rejection. Twelve non-transplant kidney biopsies (8 female, 4 male) were used as controls. Tubular chimerism was detected more frequently (69%) than endothelial chimerism (12%) in renal transplants. One of 12 control patients had tubular and endothelial chimeric cells (8%). The Y chromosome occurred in 8/12 male recipients (67%) in tubular epithelial cells and in 5/12 male recipients (42%) in endothelial cells. Double X chromosomes were detected in 3/4 female recipients in tubular epithelium. Tubular chimerism occurred more often with endothelial chimerism and capillaritis without correlation with other parameters, such as rejection. Combined Y chromosomal tubular and lymphatic endothelial chimerism correlated with T-cell mediated vascular rejection in two out of three patients (66%). Combined Y chromosomal tubular and peritubular capillary chimerism correlated with antibody mediated C4d+ rejection in one out of two patients (50%). Tubular and/or endothelial chimerism occur frequently in gender mismatched renal allografts and, when combined, this is associated with T-cell mediated rejection. © 2012 The Authors. Pathology International © 2012 Japanese Society of

  12. In vitro mouse spermatogenesis with an organ culture method in chemically defined medium.

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    Hiroyuki Sanjo

    Full Text Available We previously reported the successful induction and completion of mouse spermatogenesis by culturing neonatal testis tissues. The culture medium consisted of α-minimum essential medium (α-MEM, supplemented with Knockout serum replacement (KSR or AlbuMAX, neither of which were defined chemically. In this study, we formulated a chemically defined medium (CDM that can induce mouse spermatogenesis under organ culture conditions. It was found that bovine serum albumin (BSA purified through three different procedures had different effects on spermatogenesis. We also confirmed that retinoic acid (RA played crucial roles in the onset of spermatogonial differentiation and meiotic initiation. The added lipids exhibited weak promoting effects on spermatogenesis. Lastly, luteinizing hormone (LH, follicle stimulating hormone (FSH, triiodothyronine (T3, and testosterone (T combined together promoted spermatogenesis until round spermatid production. The CDM, however, was not able to produce elongated spermatids. It was also unable to induce spermatogenesis from the very early neonatal period, before 2 days postpartum, leaving certain factors necessary for spermatogenic induction in mice unidentified. Nonetheless, the present study provided important basic information on testis organ culture and spermatogenesis in vitro.

  13. A model to analyse the flow of an incompressible Newtonian fluid through a rigid, homogeneous, isotropic and infinite porous medium

    International Nuclear Information System (INIS)

    Gama, R.M.S. da; Sampaio, R.

    1985-01-01

    The flow of an incompressible Newtonian fluid through a rigid, homogeneous, isotropic and infinite porous medium which has a given inicial distribuition of the mentioned fluid, is analyzed. It is proposed a model that assumes that the motion is caused by concentration gradient, but it does not consider the friction between the porous medium and the fluid. We solve an onedimensional case where the mathematical problem is reduced to the solution of a non-linear hyperbolic system of differential equations, subjected to an inicial condition given by a step function, called 'Riemann Problem'. (Author) [pt

  14. Differential effect of extracellular matrix derived from papillary and reticular fibroblasts on epidermal development in vitro.

    Science.gov (United States)

    Janson, David; Rietveld, Marion; Mahé, Christian; Saintigny, Gaëlle; El Ghalbzouri, Abdoelwaheb

    2017-06-01

    Papillary and reticular fibroblasts have different effects on keratinocyte proliferation and differentiation. The aim of this study was to investigate whether these effects are caused by differential secretion of soluble factors or by differential generation of extracellular matrix from papillary and reticular fibroblasts. To study the effect of soluble factors, keratinocyte monolayer cultures were grown in papillary or reticular fibroblast-conditioned medium. To study the effect of extracellular matrix, keratinocytes were grown on papillary or reticular-derived matrix. Conditioned medium from papillary or reticular fibroblasts did not differentially affect keratinocyte viability or epidermal development. However, keratinocyte viability was increased when grown on matrix derived from papillary, compared with reticular, fibroblasts. In addition, the longevity of the epidermis was increased when cultured on papillary fibroblast-derived matrix skin equivalents compared with reticular-derived matrix skin equivalents. The findings indicate that the matrix secreted by papillary and reticular fibroblasts is the main causal factor to account for the differences in keratinocyte growth and viability observed in our study. Differences in response to soluble factors between both populations were less significant. Matrix components specific to the papillary dermis may account for the preferential growth of keratinocytes on papillary dermis.

  15. Diffraction of a Gaussian beam in a three-dimensional smoothly inhomogeneous medium: an eikonal-based complex geometrical-optics approach.

    Science.gov (United States)

    Berczynski, Pawel; Bliokh, Konstantin Yu; Kravtsov, Yuri A; Stateczny, Andrzej

    2006-06-01

    We present an ab initio account of the paraxial complex geometrical optics (CGO) in application to scalar Gaussian beam propagation and diffraction in a 3D smoothly inhomogeneous medium. The paraxial CGO deals with quadratic expansion of the complex eikonal and reduces the wave problem to the solution of ordinary differential equations of the Riccati type. This substantially simplifies the description of Gaussian beam diffraction as compared with full-wave or parabolic (quasi-optics) equations. For a Gaussian beam propagating in a homogeneous medium or along the symmetry axis in a lenslike medium, the CGO equations possess analytical solutions; otherwise, they can be readily solved numerically. As a nontrivial example we consider Gaussian beam propagation and diffraction along a helical ray in an axially symmetric waveguide medium. It is shown that the major axis of the beam's elliptical cross section grows unboundedly; it is oriented predominantly in the azimuthal (binormal) direction and does not obey the parallel-transport law.

  16. Proliferation and differentiation of bone marrow stromal cells under hypoxic conditions

    International Nuclear Information System (INIS)

    Ren Hongying; Cao Ying; Zhao, Qinjun; Li Jing; Zhou Cixiang; Liao Lianming; Jia Mingyue; Zhao Qian; Cai Huiguo; Han Zhongchao; Yang Renchi; Chen Guoqiang; Zhao, R.C.

    2006-01-01

    Low oxygen tension is a potent differentiation inducer of numerous cell types and an effective stimulus of many gene expressions. Here, we described that under 8% O 2 , bone marrow stromal cells (MSCs) exhibited proliferative and morphologic changes. The level of differentiated antigen H-2Dd and the number of G 2 /S/M phase cells increased evidently under 8% O 2 condition. Also, the proportion of wide, flattened, and epithelial-like cells (which were alkaline phosphatase staining positive) in MSCs increased significantly. When cultured in adipogenic medium, there was a 5- to 6-fold increase in the number of lipid droplets under hypoxic conditions compared with that in normoxic culture. We also demonstrated the existence of MSC differentiation under hypoxic conditions by electron microscopy. Expression of Oct4 was inhibited under 8% O 2 condition, but after adipocyte differentiation in normoxic culture and hypoxia-mimicking agents cobalt chloride (CoCl 2 ) and deferoxamine mesylate (DFX) treatments, Oct4 was still expressed in MSCs. These results indicate hypoxia accelerates MSC differentiation and hypoxia and hypoxia-mimicking agents exert different effects on MSC differentiation

  17. Computer-aided analysis of CT images for the differentiation of cerebral tumors

    International Nuclear Information System (INIS)

    Michalik, M.; Michalik, S.; Bornholdt, F.

    1988-01-01

    For the integration of CT imaging into the differential diagnostics of intracranial space occupations, the selection and description of characteristics facilitating a good discrimination of serveral classes of tumors becomes a very important task. From images of 93 patients with the most frequent brain tumors the optimal set of characteristics was determined. The four most significant characteristics for the differentiation of brain tumors are 'uptake of contrast medium by the tumor', 'deliniation of the tumor contours', 'progression of the tumor' and the 'average tumor density after administration of contrast media'. Very good results were obtained for the differentiation of menigneomas with and without anaplasia and for the differentiation of meningeomas from all other tumors examined. The differentiation of the degree of malignancy for various gliomatous tumors was difficult. An accurate reclassification with the computer program was obtained for 83.4% of all tumors. (author)

  18. A modeling of sliding joint on one-dimensional flexible medium

    International Nuclear Information System (INIS)

    Hong Difeng; Ren Gexue

    2011-01-01

    The dynamic modeling of a sliding joint on a one-dimensional medium, such as a cable or a beam, is studied in this paper. The sliding joint is implemented by positioning it at a moving node on the one-dimensional medium, which is realized by variable-length elements at either side of the joint. The variable-length element is established with an absolute nodal coordinate formulation (ANCF) in the framework of the Arbitrary Lagrange–Euler (ALE) description. The sliding of the joint is described by the increasing of the length on one side of the one-dimensional medium and a corresponding decreasing of the other side. In order to capture the discontinuity of the slopes at the position of the sliding joint, the moving node has two slopes as generalized coordinates which are equal to each other in the case of a beam but not in the case of a cable, and in order to avoid the addition–deletion constraint, the node adjacent to the moving node is added or deleted if the element is too long or too short. The governing equations for the coupled system are derived in terms of D’Alembert’s principle and the resulting equations of motion are formulated in the standard form of differential algebraic equations of multibody systems. Numerical examples are presented to validate the method proposed by comparing with analytical results which are available or are made possible by simplifying the model.

  19. Isolation and Multiple Differentiation Potential Assessment of Human Gingival Mesenchymal Stem Cells

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    Yuan Gao

    2014-11-01

    Full Text Available The aim of this study was to isolate human mesenchymal stem cells (MSCs from the gingiva (GMSCs and confirm their multiple differentiation potentials, including the odontogenic lineage. GMSCs, periodontal ligament stem cells (PDLSCs and dermal stem cells (DSCs cultures were analyzed for cell shape, cell cycle, colony-forming unit-fibroblast (CFU-F and stem cell markers. Cells were then induced for osteogenic and adipogenic differentiation and analyzed for differentiation markers (alkaline phosphatase (ALP activity, mineralization nodule formation and Runx2, ALP, osteocalcin (OCN and collagen I expressions for the osteogenic differentiation, and lipid vacuole formation and PPARγ-2 expression for the adipogenic differentiation. Besides, the odontogenic differentiation potential of GMSCs induced with embryonic tooth germ cell-conditioned medium (ETGC-CM was observed. GMSCs, PDLSCs and DSCs were all stromal origin. PDLSCs showed much higher osteogenic differentiation ability but lower adipogenic differentiation potential than DSCs. GMSCs showed the medial osteogenic and adipogenic differentiation potentials between those of PDLSCs and DSCs. GMSCs were capable of expressing the odontogenic genes after ETGC-CM induction. This study provides evidence that GMSCs can be used in tissue engineering/regeneration protocols as an approachable stem cell source.

  20. Zika virus infection dysregulates human neural stem cell growth and inhibits differentiation into neuroprogenitor cells.

    Science.gov (United States)

    Devhare, Pradip; Meyer, Keith; Steele, Robert; Ray, Ratna B; Ray, Ranjit

    2017-10-12

    The current outbreak of Zika virus-associated diseases in South America and its threat to spread to other parts of the world has emerged as a global health emergency. A strong link between Zika virus and microcephaly exists, and the potential mechanisms associated with microcephaly are under intense investigation. In this study, we evaluated the effect of Zika virus infection of Asian and African lineages (PRVABC59 and MR766) in human neural stem cells (hNSCs). These two Zika virus strains displayed distinct infection pattern and growth rates in hNSCs. Zika virus MR766 strain increased serine 139 phosphorylation of histone H2AX (γH2AX), a known early cellular response proteins to DNA damage. On the other hand, PRVABC59 strain upregulated serine 15 phosphorylation of p53, p21 and PUMA expression. MR766-infected cells displayed poly (ADP-ribose) polymerase (PARP) and caspase-3 cleavage. Interestingly, infection of hNSCs by both strains of Zika virus for 24 h, followed by incubation in astrocyte differentiation medium, induced rounding and cell death. However, astrocytes generated from hNSCs by incubation in differentiation medium when infected with Zika virus displayed minimal cytopathic effect at an early time point. Infected hNSCs incubated in astrocyte differentiating medium displayed PARP cleavage within 24-36 h. Together, these results showed that two distinct strains of Zika virus potentiate hNSC growth inhibition by different mechanisms, but both viruses strongly induce death in early differentiating neuroprogenitor cells even at a very low multiplicity of infection. Our observations demonstrate further mechanistic insights for impaired neuronal homeostasis during active Zika virus infection.

  1. Zika virus infection dysregulates human neural stem cell growth and inhibits differentiation into neuroprogenitor cells

    Science.gov (United States)

    Devhare, Pradip; Meyer, Keith; Steele, Robert; Ray, Ratna B; Ray, Ranjit

    2017-01-01

    The current outbreak of Zika virus-associated diseases in South America and its threat to spread to other parts of the world has emerged as a global health emergency. A strong link between Zika virus and microcephaly exists, and the potential mechanisms associated with microcephaly are under intense investigation. In this study, we evaluated the effect of Zika virus infection of Asian and African lineages (PRVABC59 and MR766) in human neural stem cells (hNSCs). These two Zika virus strains displayed distinct infection pattern and growth rates in hNSCs. Zika virus MR766 strain increased serine 139 phosphorylation of histone H2AX (γH2AX), a known early cellular response proteins to DNA damage. On the other hand, PRVABC59 strain upregulated serine 15 phosphorylation of p53, p21 and PUMA expression. MR766-infected cells displayed poly (ADP-ribose) polymerase (PARP) and caspase-3 cleavage. Interestingly, infection of hNSCs by both strains of Zika virus for 24 h, followed by incubation in astrocyte differentiation medium, induced rounding and cell death. However, astrocytes generated from hNSCs by incubation in differentiation medium when infected with Zika virus displayed minimal cytopathic effect at an early time point. Infected hNSCs incubated in astrocyte differentiating medium displayed PARP cleavage within 24–36 h. Together, these results showed that two distinct strains of Zika virus potentiate hNSC growth inhibition by different mechanisms, but both viruses strongly induce death in early differentiating neuroprogenitor cells even at a very low multiplicity of infection. Our observations demonstrate further mechanistic insights for impaired neuronal homeostasis during active Zika virus infection. PMID:29022904

  2. Hybrid Complexes of High and Low Molecular Weight Hyaluronans Highly Enhance HASCs Differentiation: Implication for Facial Bioremodelling

    Directory of Open Access Journals (Sweden)

    Antonietta Stellavato

    2017-11-01

    Full Text Available Background/Aims: Adipose-derived Stem Cells (ASCs are used in Regenerative Medicine, including fat grafting, recovery from local tissue ischemia and scar remodeling. The aim of this study was to evaluate hyaluronan based gel effects on ASCs differentiation and proliferation. Methods: Comparative analyses using high (H and low (L molecular weight hyaluronans (HA, hyaluronan hybrid cooperative complexes (HCCs, and high and medium cross-linked hyaluronan based dermal fillers were performed. Human ASCs were characterized by flow cytometry using CD90, CD34, CD105, CD29, CD31, CD45 and CD14 markers. Then, cells were treated for 7, 14 and 21 days with hyaluronans. Adipogenic differentiation was evaluated using Oil red-O staining and expression of leptin, PPAR-γ, LPL and adiponectin using qRT-PCR. Adiponectin was analyzed by immunofluorescence, PPAR-γ and adiponectin were analyzed using western blotting. ELISA assays for adiponectin and leptin were performed. Results: HCCs highly affected ASCs differentiation by up-regulating adipogenic genes and related proteins, that were also secreted in the culture medium. H-HA and L-HA induced a lower level of ASCs differentiation. Conclusion: HCCs-based formulations clearly enhance adipogenic differentiation and proliferation, when compared with linear HA and cross-linked hyaluronans. Injection of HCCs in subdermal fat compartment may recruit and differentiate stem cells in adipocytes, and considerably improving fat tissue renewal.

  3. Probing in-medium spin–orbit interaction with intermediate-energy heavy-ion collisions

    International Nuclear Information System (INIS)

    Xu, Jun; Li, Bao-An

    2013-01-01

    Incorporating for the first time both the spin and isospin degrees of freedom explicitly in transport model simulations of intermediate-energy heavy-ion collisions, we observe that a local spin polarization appears during collision process. Most interestingly, it is found that the nucleon spin up–down differential transverse flow is a sensitive probe of the spin–orbit interaction, providing a novel approach to probe both the density and isospin dependence of the in-medium spin–orbit coupling that is important for understanding the structure of rare isotopes and synthesis of superheavy elements

  4. Simple and complex chimera states in a nonlinearly coupled oscillatory medium

    Science.gov (United States)

    Bolotov, Maxim; Smirnov, Lev; Osipov, Grigory; Pikovsky, Arkady

    2018-04-01

    We consider chimera states in a one-dimensional medium of nonlinear nonlocally coupled phase oscillators. In terms of a local coarse-grained complex order parameter, the problem of finding stationary rotating nonhomogeneous solutions reduces to a third-order ordinary differential equation. This allows finding chimera-type and other inhomogeneous states as periodic orbits of this equation. Stability calculations reveal that only some of these states are stable. We demonstrate that an oscillatory instability leads to a breathing chimera, for which the synchronous domain splits into subdomains with different mean frequencies. Further development of instability leads to turbulent chimeras.

  5. Automatic differentiation as a tool in engineering design

    Science.gov (United States)

    Barthelemy, Jean-Francois; Hall, Laura E.

    1992-01-01

    Automatic Differentiation (AD) is a tool that systematically implements the chain rule of differentiation to obtain the derivatives of functions calculated by computer programs. AD is assessed as a tool for engineering design. The forward and reverse modes of AD, their computing requirements, as well as approaches to implementing AD are discussed. The application of two different tools to two medium-size structural analysis problems to generate sensitivity information typically necessary in an optimization or design situation is also discussed. The observation is made that AD is to be preferred to finite differencing in most cases, as long as sufficient computer storage is available; in some instances, AD may be the alternative to consider in lieu of analytical sensitivity analysis.

  6. A new numerical approximation of the fractal ordinary differential equation

    Science.gov (United States)

    Atangana, Abdon; Jain, Sonal

    2018-02-01

    The concept of fractal medium is present in several real-world problems, for instance, in the geological formation that constitutes the well-known subsurface water called aquifers. However, attention has not been quite devoted to modeling for instance, the flow of a fluid within these media. We deem it important to remind the reader that the concept of fractal derivative is not to represent the fractal sharps but to describe the movement of the fluid within these media. Since this class of ordinary differential equations is highly complex to solve analytically, we present a novel numerical scheme that allows to solve fractal ordinary differential equations. Error analysis of the method is also presented. Application of the method and numerical approximation are presented for fractal order differential equation. The stability and the convergence of the numerical schemes are investigated in detail. Also some exact solutions of fractal order differential equations are presented and finally some numerical simulations are presented.

  7. Lifespan Extension and Sustained Expression of Stem Cell Phenotype of Human Breast Epithelial Stem Cells in a Medium with Antioxidants

    Directory of Open Access Journals (Sweden)

    Kai-Hung Wang

    2016-01-01

    Full Text Available We have previously reported the isolation and culture of a human breast epithelial cell type with stem cell characteristics (Type I HBEC from reduction mammoplasty using the MSU-1 medium. Subsequently, we have developed several different normal human adult stem cell types from different tissues using the K-NAC medium. In this study, we determined whether this low calcium K-NAC medium with antioxidants (N-acetyl-L-cysteine and L-ascorbic acid-2-phosphate is a better medium to grow human breast epithelial cells. The results clearly show that the K-NAC medium is a superior medium for prolonged growth (cumulative population doubling levels ranged from 30 to 40 of normal breast epithelial cells that expressed stem cell phenotypes. The characteristics of these mammary stem cells include deficiency in gap junctional intercellular communication, expression of Oct-4, and the ability to differentiate into basal epithelial cells and to form organoid showing mammary ductal and terminal end bud-like structures. Thus, this new method of growing Type I HBECs will be very useful in future studies of mammary development, breast carcinogenesis, chemoprevention, and cancer therapy.

  8. Technical note: a pilot study using a mouse mastitis model to study differences between bovine associated coagulase-negative staphylococci.

    Science.gov (United States)

    Breyne, K; De Vliegher, S; De Visscher, A; Piepers, S; Meyer, E

    2015-02-01

    Coagulase-negative staphylococci (CNS) are a group of bacteria classified as either minor mastitis pathogens or commensal microbiota. Recent research suggests species- and even strain-related epidemiological and genetic differences within the large CNS group. The current pilot study investigated in 2 experiments whether a mouse mastitis model validated for bovine Staphylococcus aureus can be used to explore further differences between CNS species and strains. In a first dose titration experiment, a low inoculum dose of S. aureus Newbould 305 (positive control) was compared with increasing inoculum doses of a Staphylococcus chromogenes strain originating from a chronic bovine intramammary infection to a sham-inoculated mammary glands (negative control). In contrast to the high bacterial growth following inoculation with S. aureus, S. chromogenes was retrieved in very low levels at 24 h postinduction (p.i.). In a second experiment, the inflammation inflicted by 3 CNS strains was studied in mice. The host immune response induced by the S. chromogenes intramammary strain was compared with the one induced by a Staphylococcus fleurettii strain originating from cow bedding sawdust and by a S. chromogenes strain originating from a teat apex of a heifer. As expected, at 28 and 48 h p.i., low bacterial growth and local neutrophil influx in the mammary gland were induced by all CNS strains. As hypothesized, bacterial growth p.i. was the lowest for S. fleurettii compared with that induced by the 2 S. chromogenes strains, and the overall immune response established by the 3 CNS strains was less pronounced compared with the one induced by S. aureus. Proinflammatory cytokine profiling revealed that S. aureus locally induced IL-6 and IL-1β but not TNF-α, whereas, overall, CNS-inoculated glands lacked a strong cytokine host response but also induced IL-1β locally. Compared with both other CNS strains, S. chromogenes from the teat apex inflicted a more variable IL-1β response

  9. Differentiation and sarcomere formation in skeletal myocytes directly prepared from human induced pluripotent stem cells using a sphere-based culture.

    Science.gov (United States)

    Jiwlawat, Saowanee; Lynch, Eileen; Glaser, Jennifer; Smit-Oistad, Ivy; Jeffrey, Jeremy; Van Dyke, Jonathan M; Suzuki, Masatoshi

    Human induced-pluripotent stem cells (iPSCs) are a promising resource for propagation of myogenic progenitors. Our group recently reported a unique protocol for the derivation of myogenic progenitors directly (without genetic modification) from human pluripotent cells using free-floating spherical culture. Here we expand our previous efforts and attempt to determine how differentiation duration, culture surface coatings, and nutrient supplements in the medium influence progenitor differentiation and formation of skeletal myotubes containing sarcomeric structures. A long differentiation period (over 6 weeks) promoted the differentiation of iPSC-derived myogenic progenitors and subsequent myotube formation. These iPSC-derived myotubes contained representative sarcomeric structures, consisting of organized myosin and actin filaments, and could spontaneously contract. We also found that a bioengineering approach using three-dimensional (3D) artificial muscle constructs could facilitate the formation of elongated myotubes. Lastly, we determined how culture surface coating matrices and different supplements would influence terminal differentiation. While both Matrigel and laminin coatings showed comparable effects on muscle differentiation, B27 serum-free supplement in the differentiation medium significantly enhanced myogenesis compared to horse serum. Our findings support the possibility to create an in vitro model of contractile sarcomeric myofibrils for disease modeling and drug screening to study neuromuscular diseases. Copyright © 2017 International Society of Differentiation. Published by Elsevier B.V. All rights reserved.

  10. Human serum promotes osteogenic differentiation of human dental pulp stem cells in vitro and in vivo.

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    Alessandra Pisciotta

    Full Text Available Human dental pulp is a promising alternative source of stem cells for cell-based tissue engineering in regenerative medicine, for the easily recruitment with low invasivity for the patient and for the self-renewal and differentiation potential of cells. So far, in vitro culture of mesenchymal stem cells is usually based on supplementing culture and differentiation media with foetal calf serum (FCS. FCS is known to contain a great quantity of growth factors, and thus to promote cell attachment on plastic surface as well as expansion and differentiation. Nevertheless, FCS as an animal origin supplement may represent a potential means for disease transmission besides leading to a xenogenic immune response. Therefore, a significant interest is focused on investigating alternative supplements, in order to obtain a sufficient cell number for clinical application, avoiding the inconvenients of FCS use. In our study we have demonstrated that human serum (HS is a suitable alternative to FCS, indeed its addition to culture medium induces a high hDPSCs proliferation rate and improves the in vitro osteogenic differentiation. Furthermore, hDPSCs-collagen constructs, pre-differentiated with HS-medium in vitro for 10 days, when implanted in immunocompromised rats, are able to restore critical size parietal bone defects. Therefore these data indicate that HS is a valid substitute for FCS to culture and differentiate in vitro hDPSCs in order to obtain a successful bone regeneration in vivo.

  11. [Survey on the medical mycology processes to the members of the French Society for Medical Mycology].

    Science.gov (United States)

    Kauffmann-Lacroix, C; Albouy-Llaty, M; Migeot, V; Contet-Audonneau, N

    2011-09-01

    The objective of the survey was to describe the practices of clinical laboratories in terms of cultures in medical mycology. We have implemented this project within the members of the French Society for Medical Mycology (SFMM) to evaluate the analytical processes of the mycological examination in our laboratories. This preliminary study would help to suggest the future French guidelines. A questionnaire regarding the processing of mycology analysis was sent to the 227 members of the SFMM in 2009. The data involved 21 types of samples, direct microscopic examination with or without colouring and the reagents, the number of culture media, the types of media (Sabouraud, Sabouraud antibiotic, Sabouraud cycloheximide and chromogenic medium), temperature and duration of the incubation (days) and the existence of a first result before the end of the incubation period. The analytical processes were compared to an accredited laboratory according to EN ISO 15189. A great heterogeneity was observed in the 36 forms from 27 (75%) laboratories belonging to university hospitals among the 38 existing in France. As for deep samples, two microscopic exams were performed, only one was usually done. A more sensitive technique was preferred to the wet-mount for some samples. Routine samples are often inoculated on a chromogenic media. For deep samples two medium are inoculated (chromogenic media, Sabouraud and antibiotics). If the temperature of incubation is unique, 30°C was chosen. A temperature of 37°C was preferred for samples where Candida spp. is selected. When there are two temperatures of incubation, 27°C and 37°C were preferred. Each biologist can compare his proceedings to the other laboratories and to a laboratory already accredited. The question is to find the best strategies for each medical mycology specimen. They will aid the process of accreditation according to EN ISO 15189, which now applies in all laboratories in Europe. Copyright © 2011 Elsevier Masson SAS

  12. An intercomparison of medium energy cross-section codes

    International Nuclear Information System (INIS)

    Pearlstein, S.

    1988-05-01

    Five medium energy proton reaction cases are selected for benchmarking nuclear model codes. The quantities calculated are isotopic activation yields for 180 MeV protons on Al and 40-200 MeV protons on Co, and double differential neutron emission spectra from Al, Zr-90 and Pb-208 for 35, 80, 160, 318, and 800 presented consist of three types: a closed form preequilibrium plus evaporation model, an intranuclear-cascade and evaporation model, and a model relying on nuclear systematics. The characteristics of each code are described. There are orders of magnitude differences in the time for each type of code to calculate neutron emission spectra, with codes using systematics, preequilibrium and intranuclear-cascade models requiring seconds, minutes and hours, respectively. Calculations are not compared with experiment in this initial study. For double differential neutron emission spectra, there is good overall agreement in magnitude among the different types of codes at forward angles. Differences where they occur at forward angles are greatest for the mid-energy neutrons emitted. At back angles the incident energy at which the best overall agreement is obtained is 160 MeV and the material for which the best overall agreement is obtained is Al. 4 refs., 7 tabs

  13. COMPARISON OF TWO METHODS FOR THE DETECTION OF LISTERIA MONOCYTOGENES

    Directory of Open Access Journals (Sweden)

    G. Tantillo

    2013-02-01

    Full Text Available The aim of this study was to compare the performance of the conventional methods for detection of Listeria monocytogenes in food using media Oxford and ALOA (Agar Listeria acc. to Ottaviani & Agosti in according to the ISO 11290-1 to a new chromogenic medium “CHROMagar Listeria” standardized in 2005 AFNOR ( CHR – 21/1-12/01. A total of 40 pre-packed ready-to-eat food samples were examined. Using two methods six samples were found positive for Listeria monocytogenes but the medium “CHROMagar Listeria” was more selective in comparison with the others. In conclusion this study has demonstrated that isolation medium able to target specifically the detection of L. monocytogenes such as “CHROMagar Listeria” is highly recommendable because of that detection time is significantly reduced and the analysis cost is less expensive.

  14. Validity of whole slide images for scoring HER2 chromogenic in situ hybridisation in breast cancer.

    Science.gov (United States)

    Al-Janabi, Shaimaa; Horstman, Anja; van Slooten, Henk-Jan; Kuijpers, Chantal; Lai-A-Fat, Clifton; van Diest, Paul J; Jiwa, Mehdi

    2016-05-09

    Whole slide images (WSIs) have stimulated a paradigm shift from conventional to digital pathology in several applications within pathology. Due to the fact that WSIs have not yet been approved for primary diagnostics, validating their use for different diagnostic purposes is still mandatory. The aim of this study was to test the validity of WSI in assessing human epidermal growth factor receptor 2 (HER2) status in breast cancer specimens using chromogenic in situ hybridisation (CISH). Ninety-six HER2 CISH slides were scored by two observers on a light microscope (400× viewing magnification) and on WSI (40× scanning magnification, one focus plane) with a minimum of 6 weeks washout period. The concordance between digital and microscopic HER2 scores was assessed. Digitally, 93/96 cases could be assessed (96.8%). Microscopic and digital evaluation of HER2 amplification status were concordant in 68/93 cases ((73.1%, 95% CI: 0.639 -0.823), κ 0.588). CISH underscoring was most noticeable in the amplified and equivocal categories while the highest level concordance was seen in cases with a normal copy number. Additionally there was a noticeable tendency to underestimate the average HER2 scores on WSI: lower in 59 and higher in 11 cases. There was no major difference in time spent for microscopic scoring (86.9 s) and digital scoring (81.7 s). There was a reasonable concordance between microscopic scoring and WSI-based scoring of HER2 copy number of CISH slides. Nevertheless, WSIs scanned on a single focal plane are insufficient to assess HER2 gene amplification status by scoring CISH due to the noticeable tendency towards digitally underestimating the number of HER2 spots. Scanning at multiple focus planes may offer better resolution for improved digital CISH spot counting. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/

  15. Determination of HER2 gene amplification by chromogenic in situ hybridization (CISH) in archival breast carcinoma.

    Science.gov (United States)

    Zhao, Jianxin; Wu, Rina; Au, Alfred; Marquez, Abbey; Yu, Yibing; Shi, Zuorong

    2002-06-01

    To compare the efficacy of chromogenic in situ hybridization (CISH(TM)) with fluorescence in situ (FISH) hybridization and immunohistochemistry (IHC) in determination of the HER2 status in human breast cancer. HER2 gene amplification was determined on formalin-fixed paraffin-embedded (FFPE) sections of 62 invasive breast cancers by FISH and followed by CISH using a digoxigenin (DIG)-labeled HER2 DNA probe generated by Subtraction Probe Technology (SPT(TM)), and a biotin-labeled chromosome 17 centromeric (chr.17cen) probe. The sections were heat treated and enzyme digested. After in situ hybridization, the HER2 probe was detected with fluorescein (FITC)-anti-DIG for FISH, followed by peroxidase-anti-FITC and diaminobenzidine (DAB) for CISH. The chr.17cen probe was detected with peroxidase-streptavidin and DAB. For CISH application, HER2 gene copies or chromosome 17 centromeres and morphology of cells were easily visualized simultaneously with a 40x objective under bright-field microscope in hematoxylin-counterstained sections. IHC study of HER2 overexpression was performed on adjacent sections using a panel of three HER2 antibodies (TAB 250, CB11, A0485), and staining was scored according to the criteria specified in the HercepTest. HER2 gene amplification detected by CISH was visualized typically as large DAB-stained clusters or by many dots in the nucleus. FISH and CISH identified HER2 gene amplification in 19% of the tumors. Chromosome 17 polysomy was detected in 31% of the tumors. HER2 overexpression was demonstrated in 19% (TAB 250), 23% (CB11), and 36% (A0485) of the tumors. Complete concordance between the results of CISH with FISH, TAB 250, CB11, and A0485 was seen in 100%, 97%, 94%, and 84% of the cases, respectively. By permitting observation of morphology using a bright-field microscope, CISH is an accurate, practical, and economical approach to screen HER2 status in breast cancers. It is a useful methodology for confirming ambiguous IHC results.

  16. Ecto-mesenchymal stem cells from dental pulp are committed to differentiate into active melanocytes

    Directory of Open Access Journals (Sweden)

    F Paino

    2010-10-01

    Full Text Available Dental pulp stem cells (DPSCs are multipotent stem cells derived from neural crest and mesenchyme and have the capacity to differentiate into multiple cell lineages. It has already been demonstrated that DPSCs differentiate into melanocyte-like cells but only when cultivated in a specific melanocyte differentiating medium. In this study we have shown, for the first time, that DPSCs are capable of spontaneously differentiating into mature melanocytes, which display molecular and ultrastructural features of full development, including the expression of melanocyte specific markers and the presence of melanosomes up to the terminal stage of maturation. We have also compared the differentiating features of DPSCs grown in different culture conditions, following the timing of differentiation at molecular and cytochemical levels and found that in all culture conditions full development of these cells was obtained, although at different times. The spontaneous differentiating potential of these cells strongly suggests their possible applications in regenerative medicine.

  17. Tracer transfer in consolidated porous medium and fractured porous medium: experimentations and modelling

    International Nuclear Information System (INIS)

    Dalla Costa, C.

    2007-07-01

    We try to identify and model physical and chemical mechanisms governing the water flow and the solute transport in fractured consolidated porous medium. An original experimental device was built. The 'cube' consists of an idealized fractured medium reproduced by piling up consolidated porous cubes of 5 cm edge. Meanwhile, columns of the homogeneous consolidated porous medium are studied. The same anionic tracing technique is used in both cases. Using a system analysis approach, we inject concentration pulses in the device to obtain breakthrough curves. After identifying the mass balance and the residence time, we fit the CD and the MIM models to the experimental data. The MIM model is able to reproduce experimental curves of the homogeneous consolidated porous medium better than the CD model. The mobile water fraction is in accordance with the porous medium geometry. The study of the flow rate influence highlights an interference dispersion regime. It was not possible to highlight the observation length influence in this case. On the contrary, we highlight the effect of the observation scale on the fractured and porous medium, comparing the results obtained on a small 'cube' and a big 'cube'. The CD model is not satisfactory in this case. Even if the MIM model can fit the experimental breakthrough curves, it was not possible to obtain unique parameters for the set of experiments. (author)

  18. Statistical analysis of the established salary in small and medium enterprises

    Directory of Open Access Journals (Sweden)

    Yu. S. Pin’kovetskaya

    2017-01-01

    Full Text Available The aim of the study was to analyze the present regularities, specific to the employees’ salaries of aggregates of small and medium enterprises related to the three dimensional categories and located in different regions of Russia. The following tasks were solved: the indexes, characterizing average monthly salary of employees based on the mentioned enterprises were assessed, belonging to different size categories and located in each of the regions; the relations were established between the average monthly salaries of employees of aggregates of small and medium enterprises and the cost of living in all regions of the country.Preliminary results of stopwatch reading of small and medium business activities in 2015 were used as initial data. The research was based on the comparison of indexes for the entrepreneurial sector and the full range of enterprises and organizations.Modeling differentiation of salaries’ values of small and medium enterprises aggregations, as well as its relationship to the values of the subsistence level was based on the development of the density function of normal distribution. The quality of the developed models was checked according to the Kolmogorov-Smirnov, Pearson and Shapiro-Wilk criteria.The obtained results have some theoretical significance, in particular, when conducting research related to the justification of the proposed wage of employees of enterprises different in number, the formation of measures for increasing efficiency of the entrepreneurial sector activity. Density functions of normal distribution given in the paper can be used in the justification of concepts, plans and programs of developing small and medium entrepreneurship in regions and municipalities. The practical importance of research results connected with the possibility of their use by entrepreneurs directly (especially by beginners when assessing the potential of enterprise creation and definition of employees’ proposed salaries. In

  19. The Methodology Applied in DPPH, ABTS and Folin-Ciocalteau Assays Has a Large Influence on the Determined Antioxidant Potential.

    Science.gov (United States)

    Abramovič, Helena; Grobin, Blaž; Poklar, Nataša; Cigić, Blaž

    2017-06-01

    Antioxidant potential (AOP) is not only the property of the matrix analyzed but also depends greatly on the methodology used. The chromogenic radicals 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS•+), 2,2-diphenyl-1-picrylhydrazyl (DPPH•) and Folin-Ciocalteu (FC) assay were applied to estimate how the method and the composition of the assay solvent influence the AOP determined for coffee, tea, beer, apple juice and dietary supplements. Large differences between the AOP values depending on the reaction medium were observed, with the highest AOP determined mostly in the FC assay. In reactions with chromogenic radicals several fold higher values of AOP were obtained in buffer pH 7.4 than in water or methanol. The type of assay and solvent composition have similar influences on the reactivity of a particular antioxidant, either pure or as part of a complex matrix. The reaction kinetics of radicals with antioxidants in samples reveals that AOP depends strongly on incubation time, yet differently for each sample analyzed and the assay applied.

  20. Use of a novel medium, the Polymyxin Ceftazidime Oxford Medium, for isolation of Listeria monocytogenes from raw or non-pasteurized foods.

    Science.gov (United States)

    Martínez-Gonzáles, N E; Martínez-Chávez, L; Cabrera-Díaz, E; Martínez-Cárdenas, C; Gutiérrez-González, P; Castillo, A

    2016-05-01

    Polymyxin Ceftazidime Oxford Medium (PCOM), a novel selective and differential plating medium for Listeria monocytogenes was compared with Modified Oxford Agar (MOX) for efficacy to isolate L. monocytogenes and other Listeria spp. naturally present in non-pasteurized Mexican-style cheese (n = 50), non-pasteurized fresh squeezed orange juice (n = 50), raw beef chunks (n = 36), and fresh cabbage (n = 125). Samples were collected from retail markets and farms in Mexico and tested following the US Department of Agriculture enrichment technique. Listeria spp. were isolated from 23.4% of analyzed samples, and from those, 75.0% corresponded to raw beef chunks, 38.0% to non-pasteurized Mexican-style cheese, and 30.0% to fresh squeezed orange juice. No Listeria spp. were isolated from fresh cabbage samples. L. monocytogenes was recovered from 15.3% of food samples analyzed. Non-pasteurized Mexican-style cheese showed the highest proportion of L. monocytogenes positive samples (36.0%), followed by orange juice (26.0%) and raw beef (25.0%). The frequency of isolation of Listeria spp. and L. monocytogenes was not different (P > 0.05) between PCOM and MOX. The advantages of using PCOM when comparing to MOX, include the easier way to identify Listeria species, the lower cost per plate and the availability of its ingredients for Latin-American countries. Copyright © 2015 Elsevier Ltd. All rights reserved.

  1. On the radiative transfer problem in a spherical medium subject to Fresnel's reflective boundary conditions

    International Nuclear Information System (INIS)

    Mohammed, M.H.H.

    2012-01-01

    Radiation transfer problem for anisotropic scattering in a spherical homogeneous, turbid medium with angular dependent (specular) and diffuse reflecting boundary is considered. The angular dependent reflectivity of the boundary is considered as Fresnel's reflection probability function. The solution of the problem containing an energy source in a medium of specular and diffuse reflecting boundaries is given in terms of the solution of the source-free problem. The source-free problem for anisotropic scattering through a homogeneous solid sphere and two concentric spheres is solved by using the Pomraning- Eddington approximation method. This method transform the integro-differential equation into two differential equations for the radiance g (x) and net flux q (x) which has an analytical solution in terms of the modified Bessel function. Two different weight functions are used to verify the boundary conditions and so, find the solution constants. The partial heat fluxes at the boundaries of a solid sphere and spherical shell of transparent and reflecting boundaries are calculated. The media are taken with or without internal black-body radiation. The calculations are carried out for various values of refractive index and different radii. The results are compared with those of the Galerkin technique

  2. The Interstellar Medium

    CERN Document Server

    Lequeux, James

    2005-01-01

    Describing interstellar matter in our galaxy in all of its various forms, this book also considers the physical and chemical processes that are occurring within this matter. The first seven chapters present the various components making up the interstellar matter and detail the ways that we are able to study them. The following seven chapters are devoted to the physical, chemical and dynamical processes that control the behaviour of interstellar matter. These include the instabilities and cloud collapse processes that lead to the formation of stars. The last chapter summarizes the transformations that can occur between the different phases of the interstellar medium. Emphasizing methods over results, "The Interstellar Medium" is written for graduate students, for young astronomers, and also for any researchers who have developed an interest in the interstellar medium.

  3. Delta-like 1/fetal antigen-1 (Dlk1/FA1) is a novel regulator of chondrogenic cell differentiation via inhibition of the Akt kinase-dependent pathway

    DEFF Research Database (Denmark)

    Chen, Li; Qanie, Diyako; Jafari, Abbas

    2011-01-01

    Delta-like 1 (Dlk1, also known as fetal antigen-1, FA1) is a member of Notch/Delta family that inhibits adipocyte and osteoblast differentiation; however, its role in chondrogenesis is still not clear. Thus, we overexpressed Dlk1/FA1 in mouse embryonic ATDC5 cells and tested its effects...... on chondrogenic differentiation. Dlk1/FA1 inhibited insulin-induced chondrogenic differentiation as evidenced by reduction of cartilage nodule formation and gene expression of aggrecan, collagen Type II and X. Similar effects were obtained either by using Dlk1/FA1-conditioned medium or by addition of a purified......, secreted, form of Dlk1 (FA1) directly to the induction medium. The inhibitory effects of Dlk1/FA1 were dose-dependent and occurred irrespective of the chondrogenic differentiation stage: proliferation, differentiation, maturation, or hypertrophic conversion. Overexpression or addition of the Dlk1/FA1...

  4. Concept medium programme

    DEFF Research Database (Denmark)

    Bjerrum, Peter

    2005-01-01

    The present essay is an attempt to determine the architectural project of the 21st century in relation to a modern conception of space as the medium of architecture, and of sociality as its program......The present essay is an attempt to determine the architectural project of the 21st century in relation to a modern conception of space as the medium of architecture, and of sociality as its program...

  5. Branding and outcomes in small and medium-sized enterprises (SMEs)

    DEFF Research Database (Denmark)

    Odoom, Raphael; Narteh, Bedman; Rand, John

    2017-01-01

    The study investigates the relationships of enterprise resources and branding capabilities with branding efforts and branding benefits. It examines the differential effect of physical resources and branding capabilities on enterprises’ branding efforts and outcomes. Empirical data for the study...... were drawn from 304 small and medium-sized enterprises (SMEs) in Ghana. The hypothesized relationships were analyzed using Structural Equation Modeling. The study found that resources and capabilities possessions might not be enough to produce the optimum branding benefits for enterprises. A better...... result, however, emerges when these resources and capabilities are integrated with well-coordinated branding efforts of the enterprises. The study offers several implications for managers of small businesses based on the findings of the study....

  6. Computer tomographic diagnosis and differential diagnosis of juvenile angiofibromas and angiomatous polyps

    International Nuclear Information System (INIS)

    Irnberger, T.

    1985-01-01

    Juvenile angiofibromas (JAF) and angiomatous polyps (AP) are vascular histologically benign tumors presenting with an identical clinical triad. Both lesions can be differentiated by their typical geographic pattern of growth, different pattern of tumor extension and their highly characteristic angiographic and angiocomputertomographic appearance. Time-density curves were of no use in differentiating the big variety of hypervascular lesions from each other. CT images of the vascular tumor phase after bolus injection of contrast medium however showed interesting pathomorphological details, which were extremely useful in differentiating JAF from AP and other vascular tumors. It is our opinion, that invasive angiography for diagnosing JAF has been surpassed, but is further essential for mapping of tumor perfusion, feeding vessels and praeoperative embolization therapy. (orig.) [de

  7. Synthesis of glycosaminoglycans by undifferentiated and differentiated HT29 human colonic cancer cells.

    Science.gov (United States)

    Simon-Assmann, P; Bouziges, F; Daviaud, D; Haffen, K; Kedinger, M

    1987-08-15

    Among the extracellular matrix components which have been suggested to be involved in developmental and neoplastic changes are glycosaminoglycans (GAGs). To try to correlate their amount and nature with the process of enterocytic differentiation, we studied glycosaminoglycan synthesis of human colonic adenocarcinoma cells (HT29 cell line) by [3H]glucosamine and [35S]sulfate incorporation. Enterocytic differentiation of the cells obtained in a sugar-free medium (for review, see A. Zweibaum et al. In: Handbook of Physiology. Intestinal Transport of the Gastrointestinal System, in press, 1987) resulted in a marked increase in total incorporation of labeled precursors (20-fold for [3H]glucosamine, 4.5-fold for [35S]sulfate) as well as in uronic acid content (5-fold); most of the synthesized GAGs were found associated with the cell pellet. Chromatographic and electrophoretic analysis of the labeled GAGs revealed that undifferentiated cells synthesized and secreted hyaluronic acid, heparan sulfate, and one class of chondroitin sulfate. Differentiation of HT29 cells because associated with the synthesis of an additional class of chondroitin sulfate (CS4) concomitant to a decrease in heparan sulfate which is no longer found secreted in the medium. Furthermore, the charge density of this latter GAG component varied as assessed by a shift of its affinity on ion-exchange chromatography.

  8. Chromogenic in situ hybridization is a reliable assay for detection of ALK rearrangements in adenocarcinomas of the lung.

    Science.gov (United States)

    Schildhaus, Hans-Ulrich; Deml, Karl-Friedrich; Schmitz, Katja; Meiboom, Maren; Binot, Elke; Hauke, Sven; Merkelbach-Bruse, Sabine; Büttner, Reinhard

    2013-11-01

    Reliable detection of anaplastic lymphoma kinase (ALK) rearrangements is a prerequisite for personalized treatment of lung cancer patients, as ALK rearrangements represent a predictive biomarker for the therapy with specific tyrosine kinase inhibitors. Currently, fluorescent in situ hybridization (FISH) is considered to be the standard method for assessing formalin-fixed and paraffin-embedded tissue for ALK inversions and translocations. However, FISH requires a specialized equipment, the signals fade rapidly and it is difficult to detect overall morphology and tumor heterogeneity. Chromogenic in situ hybridization (CISH) has been successfully introduced as an alternative test for the detection of several genetic aberrations. This study validates a newly developed ALK CISH assay by comparing FISH and CISH signal patterns in lung cancer samples with and without ALK rearrangements. One hundred adenocarcinomas of the lung were included in this study, among them 17 with known ALK rearrangement. FISH and CISH were carried out and evaluated according to the manufacturers' recommendations. For both assays, tumors were considered positive if ≥15% of tumor cells showed either isolated 3' signals or break-apart patterns or a combination of both. A subset of tumors was exemplarily examined by using a novel EML4 (echinoderm microtubule-associated protein-like 4) CISH probe. Red, green and fusion CISH signals were clearcut and different signal patterns were easily recognized. The percentage of aberrant tumor cells was statistically highly correlated (PCISH. On the basis of 86 samples that were evaluable by ALK CISH, we found a 100% sensitivity and 100% specificity of this assay. Furthermore, EML4 rearrangements could be recognized by CISH. CISH is a highly reliable, sensitive and specific method for the detection of ALK gene rearrangements in pulmonary adenocarcinomas. Our results suggest that CISH might serve as a suitable alternative to FISH, which is the current gold

  9. Proliferation and differentiation of bone marrow stromal cells under hypoxic conditions

    Energy Technology Data Exchange (ETDEWEB)

    Hongying, Ren; Huiguo, Cai; Zhongchao, Han; Renchi, Yang; Zhao, Qinjun [State Key Lab of Experimental Hematology, Institute of Hematology and Blood Disease Hospital, Chinese Academy of Medical Sciences and Peking Union of Medical College, Tianjin (China); Ying, Cao; Jing, Li [Institute of Basic Medical Sciences and School of Basic Medicine, Center of Excellence in Tissue Engineering, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing (China); Cixiang, Zhou [Health Science Center, Shanghai Institutes of Biological Sciences, Chinese Academy of Science-SSMU, Shanghai (China); Lianming, Liao; Mingyue, Jia [Institute of Basic Medical Sciences and School of Basic Medicine, Center of Excellence in Tissue Engineering, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing (China); Qian, Zhao [Health Science Center, Shanghai Institutes of Biological Sciences, Chinese Academy of Science-SSMU, Shanghai (China); Guoqiang, Chen [Health Science Center, Shanghai Institutes of Biological Sciences, Chinese Academy of Science-SSMU, Shanghai (China); Zhao, R C [State Key Lab of Experimental Hematology, Institute of Hematology and Blood Disease Hospital, Chinese Academy of Medical Sciences and Peking Union of Medical College, Tianjin (China); [Institute of Basic Medical Sciences and School of Basic Medicine, Center of Excellence in Tissue Engineering, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing (China)]. E-mail: chunhuaz@public.tpt.tj.cn

    2006-08-18

    Low oxygen tension is a potent differentiation inducer of numerous cell types and an effective stimulus of many gene expressions. Here, we described that under 8% O{sub 2}, bone marrow stromal cells (MSCs) exhibited proliferative and morphologic changes. The level of differentiated antigen H-2Dd and the number of G{sub 2}/S/M phase cells increased evidently under 8% O{sub 2} condition. Also, the proportion of wide, flattened, and epithelial-like cells (which were alkaline phosphatase staining positive) in MSCs increased significantly. When cultured in adipogenic medium, there was a 5- to 6-fold increase in the number of lipid droplets under hypoxic conditions compared with that in normoxic culture. We also demonstrated the existence of MSC differentiation under hypoxic conditions by electron microscopy. Expression of Oct4 was inhibited under 8% O{sub 2} condition, but after adipocyte differentiation in normoxic culture and hypoxia-mimicking agents cobalt chloride (CoCl{sub 2}) and deferoxamine mesylate (DFX) treatments, Oct4 was still expressed in MSCs. These results indicate hypoxia accelerates MSC differentiation and hypoxia and hypoxia-mimicking agents exert different effects on MSC differentiation.

  10. Effect of ionizing radiation on the differentiation of neural stem cells

    International Nuclear Information System (INIS)

    Liu Ping; Tu Yu

    2010-01-01

    In order to investigate the effect of ionizing radiation on neural stem cells differentiation, we cultured neural stem cells of newborn rat in serum-free media containing EGF or bFGF. The neural stem cells were divided into 4 groups, which were irradiated by γ-rays with doses of 0, 0.5, 1, and 2 Gy. The irradiated cells were cultured under the same condition for 7 days, and the nestin content of neural stem cell was detected by immunofluorescence. The same method was carried out with irradiated cells in the culture medium after removing EGF, bFGF for 7 days, NSE and GFAP expression content and nestin were also detected by immunofluorescence. It has been found that the irradiated neural stem cells can express less nestin and differentiate more neurons compared to that of control group. Results show that ionizing radiation can induce the differentiation of the neural stem cells and make the neural stem cells differentiate more neuron. (authors)

  11. Effect of cell density on adipogenic differentiation of mesenchymal stem cells

    International Nuclear Information System (INIS)

    Lu, Hongxu; Guo, Likun; Wozniak, Michal J.; Kawazoe, Naoki; Tateishi, Tetsuya; Zhang, Xingdong; Chen, Guoping

    2009-01-01

    The effect of cell density on the adipogenic differentiation of human bone marrow-derived mesenchymal stem cells (MSCs) was investigated by using a patterning technique to induce the formation of a cell density gradient on a micropatterned surface. The adipogenic differentiation of MSCs at a density gradient from 5 x 10 3 to 3 x 10 4 cells/cm 2 was examined. Lipid vacuoles were observed at all cell densities after 1-3 weeks of culture in adipogenic differentiation medium although the lipid vacuoles were scarce at the low cell density and abundant at the high cell density. Real-time RT-PCR analysis showed that adipogenesis marker genes encoding peroxisome proliferator-activated receptor γ2 (PPARγ2), lipoprotein lipase (LPL), and fatty acid binding protein-4 (FABP4) were detected in the MSCs cultured at all cell densities. The results suggest that there was no apparent effect of cell density on the adipogenic differentiation of human MSCs.

  12. Peristaltic Flow of Carreau Fluid in a Rectangular Duct through a Porous Medium

    Directory of Open Access Journals (Sweden)

    R. Ellahi

    2012-01-01

    Full Text Available We have examined the peristaltic flow of Carreau fluid in a rectangular channel through a porous medium. The governing equations of motion are simplified by applying the long wavelength and low Reynolds number approximations. The reduced highly nonlinear partial differential equations are solved jointly by homotopy perturbation and Eigen function expansion methods. The expression for pressure rise is computed numerically by evaluating the numerical integration. The physical features of pertinent parameters have been discussed by plotting graphs of velocity, pressure rise, pressure gradient, and stream functions.

  13. Differentiation strategies and winery financial performance: An empirical investigation

    Directory of Open Access Journals (Sweden)

    Sandra K. Newton

    2015-12-01

    Full Text Available This investigation into small-to-medium sized wine businesses empirically tests linkages among differentiation strategies and financial performance over time. Using a two-by-two model, we examine the impact of differentiation strategies on profitability and growth. Financial and operational data from a proprietary database of 71 United States wineries, encompassing five continuous years (2006–2010, provide longitudinal robustness. Management decisions regarding resources and capabilities are used to cluster the sample firms into a two-by-two differentiation strategy model. Those wineries sourcing over 50% estate grapes and distributing over 50% direct-to-consumer have higher gross margins compared to other clusters. Direct-to-consumer distribution decisions impact growth. Results of this research indicate that distribution channel choice-direct-to-consumer-positively impacts gross profit margin and winery growth rates. Supply chain choice-sourcing estate grapes also positively impacts gross profit margin. This study uses reported financial data that have not been made available to researchers.

  14. Application of Medium and Seasonal Flood Forecasts for Agriculture Damage Assessment

    Science.gov (United States)

    Fakhruddin, Shamsul; Ballio, Francesco; Menoni, Scira

    2015-04-01

    Early warning is a key element for disaster risk reduction. In recent decades, major advancements have been made in medium range and seasonal flood forecasting. This progress provides a great opportunity to reduce agriculture damage and improve advisories for early action and planning for flood hazards. This approach can facilitate proactive rather than reactive management of the adverse consequences of floods. In the agricultural sector, for instance, farmers can take a diversity of options such as changing cropping patterns, applying fertilizer, irrigating and changing planting timing. An experimental medium range (1-10 day) and seasonal (20-25 days) flood forecasting model has been developed for Thailand and Bangladesh. It provides 51 sets of discharge ensemble forecasts of 1-10 days with significant persistence and high certainty and qualitative outlooks for 20-25 days. This type of forecast could assist farmers and other stakeholders for differential preparedness activities. These ensembles probabilistic flood forecasts have been customized based on user-needs for community-level application focused on agriculture system. The vulnerabilities of agriculture system were calculated based on exposure, sensitivity and adaptive capacity. Indicators for risk and vulnerability assessment were conducted through community consultations. The forecast lead time requirement, user-needs, impacts and management options for crops were identified through focus group discussions, informal interviews and community surveys. This paper illustrates potential applications of such ensembles for probabilistic medium range and seasonal flood forecasts in a way that is not commonly practiced globally today.

  15. Conditioned media from differentiating craniofacial bone marrow stromal cells influence mineralization and proliferation in periodontal ligament stem cells.

    Science.gov (United States)

    Jin, Zhenyu; Feng, Yuan; Liu, Hongwei

    2016-10-01

    Previous reports have mainly focused on the behavioral responses of human periodontal ligament stem cells (hPDLSCs) in interaction with tibia bone marrow stromal cells (BMSCs). However, there is little study on the biologic features of hPDLSCs under the induction of maxilla BMSCs (M-BMSCs) at different phases of osteogenic differentiation. We hypothesized that M-BMSCs undergoing osteogenic differentiation acted on the proliferation, differentiation, and bone-forming capacity of hPDLSCs. In this paper, primary hPDLSCs and human M-BMSCs (hM-BMSCs) were expanded in vitro. After screening of surface markers for characterization, hPDLSCs were cocultured with different phases of differentiating hM-BMSCs. Cell proliferation and alkaline phosphatase activity were examined, and mineralization-associated markers such as osteocalcin and runt-related transcription factor 2 of hPDLSCs in coculture with uninduced/osteoinduced hM-BMSCs were evaluated. hPDLSCs in hM-BMSCs-conditioned medium (hM-BMSCs-CM) group showed a reduction in proliferation compared with untreated hPDLSCs, while osteoinduced hM-BMSCs for 10 day-conditioned medium (hM-BMSCs-CM-10ds) and osteoinduced hM-BMSCs for 15 day-conditioned medium (hM-BMSCs-CM-15ds) enhance the proliferation of hPDLSCs. hM-BMSCs of separate differentiation stages temporarily inhibited osteogenesis of hPDLSCs in the early days. Upon extending time periods, uninduced/osteoinduced hM-BMSCs markedly enhanced osteogenesis of hPDLSCs to different degrees. The transplantation results showed hM-BMSCs-CM-15ds treatment promoted tissue regeneration to generate cementum/periodontal ligament-like structure characterized by hard-tissue formation. This research supported the notion that hM-BMSCs triggered osteogenesis of hPDLSCs suggesting important implications for periodontal engineering.

  16. The Effects of Sertoli Cells Condition Medium and Retinoic Acid on the Number of Colonies of Bone Marrow Mesenchymal Stem Cells

    Directory of Open Access Journals (Sweden)

    Maryam Salem

    2017-04-01

    Full Text Available Background & objectives: According to importance of bone marrow mesenchymal stem cells in production of different cell lines, transplantation of these cells are used for treatment of many different diseases during cell therapy. Viability and proliferation of these cells after transplantation are very important. Since infertility is as public health problem in men and women, the scientists attempt to produce germ cells from differentiation of stem cells. It is supposed to use these cells for treatment of different illnesses especially for men with lack of germ cells in testes in future. However, in using stem cells for cell therapy the culture medium should be designed to increase the number of cells and efficiency of transplantation and to guarantee the health of the cells in terms of DNA damage. This study designed a suitable culture medium in order to increase the number of colonies and decrease the cell injuries. Methods: In this study mesenchymal stem cells isolated from bone marrow of mice and exposed to retinoic acid (RA with concentration of 10-6 M and Sertoli cells condition medium. Since mesenchymal stem cells (MSCs produce fibroblastic colonies so the number of colonies was counted every 3 days after culture (days of 2, 5, 8, 11, and 15 under inverted microscope. The staining of ethidium bromide-acridine orange was also done for determination of apoptotic nucleus in days of 10 and 15 after culture. Results: The results showed that the effects of retinoic acid on grow and viability of MSCs is related to the time. It seems that RA increased the proliferation of the cells and the number of colonies increased in low time but the apoptotic cells elevated with increasing the time of culture. Condition medium of Sertoli cells also increased the proliferation of bone marrow stem cells. Conclusion: According to proliferative properties of condition medium, it seems that using condition medium together with RA is better than RA alone for

  17. Tattoo: a multifaceted medium of communication

    Directory of Open Access Journals (Sweden)

    Christian Wymann

    2010-11-01

    Full Text Available This article suggests the systems theoretical distinction of form/medium as a useful tool for distinguishing social phenomena that might look as if they stem from the same process. This is shown to be the case for the tattoo and tattooing. The tattoo is conceived as a medium of communication through which different forms of communication emerge. Tattooing is one of these forms of communication that shapes the medium in a particular way. The current article sheds a special light on its intricate, communicational constellation, for which the concept of parallax is suggested. Law, medicine and cosmetics as other forms of communication use the medium of tattoo in their own way as well. The form/medium distinction allows us to grasp these different forms of communication, while it shows that they share the tattoo as medium. The article’s ultimate goal is to illustrate that the tattoo figures as a multifaceted medium of communication.

  18. Portland cement induces human periodontal ligament cells to differentiate by upregulating miR-146a

    Directory of Open Access Journals (Sweden)

    Min-Ching Wang

    2018-04-01

    Full Text Available Background/Purpose: Bioaggregates such as Portland cement (PC can be an economical alternative for mineral trioxide aggregate (MTA with additional benefit of less discoloration. MTA has been known to induce differentiations of several dental cells. MicroRNAs are important regulators of biological processes, including differentiation, physiologic homeostasis, and disease progression. This study is to explore how PC enhances the differentiation of periodontal ligament (PDL cells in microRNAs level. Methods: PDL cells were cultured in a regular PC- or MTA-conditioned medium or an osteoinduction medium (OIM. Alizarin red staining was used to evaluate the extent of mineralization. Transfection of microRNA mimics induced exogenous miR-31 and miR-146a expression. The expression of microRNAs and differentiation markers was assayed using reverse-transcriptase polymerase chain reaction. Results: PC enhanced the mineralization of PDL cells in a dose-dependent manner in the OIM. Exogenous miR-31 and miR-146a expression upregulated alkaline phosphatase (ALP, bone morphogenic protein (BMP, and dentin matrix protein 1 (DMP1 expression. However, miR-31 and miR-146a modulates cementum protein 1 (CEMP1 expression in different ways. PC also enhanced ALP and BMP but attenuated CEMP1 in the OIM. Although the OIM or PC treatment upregulated miR-21, miR-29b, and miR-146a, only miR-146a was able to be induced by PC in combination with OIM. Conclusion: This study demonstrated that PC enhances the differentiation of PDL cells, especially osteogenic through miR-146a upregulation. In order to control the ankylosis after regenerative endodontics with the usage of bioaggregates, further investigations to explore these differentiation mechanisms in the miRNA level may be needed. Keywords: Portland cement, Bioaggregate, miR-146a, Osteogenic differentiation, Periodontal ligament (PDL

  19. Culture medium modulates the behaviour of human dental pulp-derived cells: Technical Note

    Directory of Open Access Journals (Sweden)

    S Lopez-Cazaux

    2006-02-01

    Full Text Available In vitro approaches have extensively been developed to study reparative dentinogenesis. While dental pulp is a source of unidentified progenitors able to differentiate into odontoblast-like cells, we investigated the effect of two media; MEM (1.8mM Ca and 1mM Pi and RPMI 1640 (0.8mM Ca and 5mM Pi on the behaviour of human dental pulp cells. Our data indicate that MEM significantly increased cell proliferation and markedly enhanced the proportion of -smooth muscle actin positive cells, which represent a putative source of progenitors able to give rise to odontoblast-like cells. In addition, MEM strongly stimulated alkaline phosphatase activity and was found to induce expression of transcripts encoding dentin sialophosphoprotein, an odontoblastic marker, without affecting that of parathyroid hormone/parathyroid hormone related protein-receptor and osteonectin. In conclusion, these observations demonstrate that not only proliferation but also differentiation into odontoblast-like cells was induced by rich calcium and poor phosphate medium (MEM as compared to RPMI 1640. This study provides important data for the determination of the optimal culture conditions allowing odontoblast-like differentiation in human pulp cell culture.

  20. Oral contrast medium in PET/CT: should you or shouldn't you?

    International Nuclear Information System (INIS)

    Groves, Ashley M.; Kayani, Irfan; Dickson, John C.; Townsend, Caroline; Croasdale, Ian; Syed, Rizwan; Nagabushan, Nagesh; Hain, Sharon F.; Ell, Peter J.; Bomanji, Jamshed B.

    2005-01-01

    It has been suggested that the use of computed tomography (CT) positive contrast agents has led to attenuation-induced artefacts on 18 F-fluorodeoxyglucose positron emission tomography ( 18 F-FDG PET/CT) systems. Consequently, centres may withhold the use of such agents. Whilst there is theoretical evidence to support the aforementioned claim, the clinical relevance of the induced artefacts has not been widely established. Moreover, the potential benefits of bowel enhancement on PET/CT have yet to be formally evaluated. We therefore prospectively examined PET/CT studies to assess whether the use of oral contrast medium induces clinically relevant artefacts and whether the use of these agents is diagnostically helpful. Over a 2-month period, 18 F-FDG PET/CT images were prospectively reviewed from 200 patients following Gastrografin administration 2 h prior to examination. Both a radiologist and a nuclear medicine physician reviewed the images for contrast medium-mediated clinically relevant artefacts. Artefacts were sought on the CT attenuation-corrected images and were compared with the appearance on non-attenuated-corrected images. The number of examinations in which the oral contrast aided image interpretation was also noted. There were no oral contrast medium-induced clinically significant artefacts. In 38 of the 200 patients, oral contrast aided image interpretation (owing to differentiation of mass/node from bowel, discrimination of intestinal wall from lumen or definition of the anatomy of a relevant site). In 33 of these 38 patients, the anatomical site of interest was the abdomen/pelvis. The use of oral contrast medium in 18 F-FDG PET studies should not be withheld as it improves image interpretation and does not produce clinically significant artefacts. (orig.)

  1. Attachment, Growth, and Detachment of Human Mesenchymal Stem Cells in a Chemically Defined Medium

    Directory of Open Access Journals (Sweden)

    Denise Salzig

    2016-01-01

    Full Text Available The manufacture of human mesenchymal stem cells (hMSCs for clinical applications requires an appropriate growth surface and an optimized, preferably chemically defined medium (CDM for expansion. We investigated a new protein/peptide-free CDM that supports the adhesion, growth, and detachment of an immortalized hMSC line (hMSC-TERT as well as primary cells derived from bone marrow (bm-hMSCs and adipose tissue (ad-hMSCs. We observed the rapid attachment and spreading of hMSC-TERT cells and ad-hMSCs in CDM concomitant with the expression of integrin and actin fibers. Cell spreading was promoted by coating the growth surface with collagen type IV and fibronectin. The growth of hMSC-TERT cells was similar in CDM and serum-containing medium whereas the lag phase of bm-hMSCs was prolonged in CDM. FGF-2 or surface coating with collagen type IV promoted the growth of bm-hMSCs, but laminin had no effect. All three cell types retained their trilineage differentiation capability in CDM and were detached by several enzymes (but not collagenase in the case of hMSC-TERT cells. The medium and coating did not affect detachment efficiency but influenced cell survival after detachment. CDM combined with cell-specific surface coatings and/or FGF-2 supplements is therefore as effective as serum-containing medium for the manufacture of different hMSC types.

  2. Tlx3 exerts context-dependent transcriptional regulation and promotes neuronal differentiation from embryonic stem cells

    OpenAIRE

    Kondo, Takako; Sheets, Patrick L.; Zopf, David A.; Aloor, Heather L.; Cummins, Theodore R.; Chan, Rebecca J.; Hashino, Eri

    2008-01-01

    The T cell leukemia 3 (Tlx3) gene has been implicated in specification of glutamatergic sensory neurons in the spinal cord. In cranial sensory ganglia, Tlx3 is highly expressed in differentiating neurons during early embryogenesis. To study a role of Tlx3 during neural differentiation, mouse embryonic stem (ES) cells were transfected with a Tlx3 expression vector. ES cells stably expressing Tlx3 were grown in the presence or absence of a neural induction medium. In undifferentiated ES cells, ...

  3. Is cyberbullying worse than traditional bullying? Examining the differential roles of medium, publicity, and anonymity for the perceived severity of bullying.

    Science.gov (United States)

    Sticca, Fabio; Perren, Sonja

    2013-05-01

    Cyberbullying, a modern form of bullying performed using electronic forms of contact (e.g., SMS, MMS, Facebook, YouTube), has been considered as being worse than traditional bullying in its consequences for the victim. This difference was mainly attributed to some specific aspect that are believed to distinguish cyberbullying from traditional bullying: an increased potential for a large audience, an increased potential for anonymous bullying, lower levels of direct feedback, decreased time and space limits, and lower levels of supervision. The present studies investigated the relative importance of medium (traditional vs. cyber), publicity (public vs. private), and bully's anonymity (anonymous vs. not anonymous) for the perceived severity of hypothetical bullying scenarios among a sample of Swiss seventh- and eight-graders (study 1: 49% female, mean age = 13.7; study 2: 49% female, mean age = 14.2). Participants ranked a set of hypothetical bullying scenarios from the most severe one to the least severe one. The scenarios were experimentally manipulated based on the aspect of medium and publicity (study 1), and medium and anonymity (study 2). Results showed that public scenarios were perceived as worse than private ones, and that anonymous scenarios were perceived as worse than not anonymous ones. Cyber scenarios generally were perceived as worse than traditional ones, although effect sizes were found to be small. These results suggest that the role of medium is secondary to the role of publicity and anonymity when it comes to evaluating bullying severity. Therefore, cyberbullying is not a priori perceived as worse than traditional bullying. Implications of the results for cyberbullying prevention and intervention are discussed.

  4. On the analysis of a complex differential game using artificial intelligence techniques [military systems

    NARCIS (Netherlands)

    Shinar, J.; Siegel, A.W.

    1988-01-01

    The investigation is motivated by the dynamic conflict in an air-to-air combat between two aggressive aircraft, both equipped with medium-range guided missiles. It is a two-target differential game with two independent pursuit-evasion games. A description is given of the analysis of the conflict by

  5. Effective medium theory for anisotropic metamaterials

    KAUST Repository

    Zhang, Xiujuan

    2015-01-20

    Materials with anisotropic material parameters can be utilized to fabricate many fascinating devices, such as hyperlenses, metasolids, and one-way waveguides. In this study, we analyze the effects of geometric anisotropy on a two-dimensional metamaterial composed of a rectangular array of elliptic cylinders and derive an effective medium theory for such a metamaterial. We find that it is possible to obtain a closed-form analytical solution for the anisotropic effective medium parameters, provided the aspect ratio of the lattice and the eccentricity of the elliptic cylinder satisfy certain conditions. The derived effective medium theory not only recovers the well-known Maxwell-Garnett results in the quasi-static regime, but is also valid beyond the long-wavelength limit, where the wavelength in the host medium is comparable to the size of the lattice so that previous anisotropic effective medium theories fail. Such an advance greatly broadens the applicable realm of the effective medium theory and introduces many possibilities in the design of structures with desired anisotropic material characteristics. A real sample of a recently theoretically proposed anisotropic medium, with a near-zero index to control the flux, is achieved using the derived effective medium theory, and control of the electromagnetic waves in the sample is clearly demonstrated.

  6. Study of the sensitivity of the radiation transport problem in a scattering medium; Estudo da sensibilidade do problema de transporte de radiacao em meio espalhador

    Energy Technology Data Exchange (ETDEWEB)

    Nunes, Rogerio Chaffin

    2002-03-15

    In this work, the system of differential equations obtained by the angular approach of the two-dimensional transport equation by the discrete ordinates method is solved through the formulation of finite elements with the objective of investigating the sensitivity of the outgoing flux of radiation with the incoming flux and the properties of absorption and scattering of the medium. The variational formulation for the system of differential equations of second order with the generalized boundary conditions of Neumann (third type) allows an easy implementation of the method of the finite elements with triangular mesh and approximation space of first order. The geometry chosen for the simulations is a circle with a non homogeneous circular form in its interior. The mapping of Dirichlet-Neumann is studied through various simulations involving the incoming flux, the outgoing flux and the properties of the medium. (author)

  7. Differential equations governing slip-induced pore-pressure fluctuations in a water-saturated granular medium

    Science.gov (United States)

    Iverson, R.M.

    1993-01-01

    Macroscopic frictional slip in water-saturated granular media occurs commonly during landsliding, surface faulting, and intense bedload transport. A mathematical model of dynamic pore-pressure fluctuations that accompany and influence such sliding is derived here by both inductive and deductive methods. The inductive derivation shows how the governing differential equations represent the physics of the steadily sliding array of cylindrical fiberglass rods investigated experimentally by Iverson and LaHusen (1989). The deductive derivation shows how the same equations result from a novel application of Biot's (1956) dynamic mixture theory to macroscopic deformation. The model consists of two linear differential equations and five initial and boundary conditions that govern solid displacements and pore-water pressures. Solid displacements and water pressures are strongly coupled, in part through a boundary condition that ensures mass conservation during irreversible pore deformation that occurs along the bumpy slip surface. Feedback between this deformation and the pore-pressure field may yield complex system responses. The dual derivations of the model help explicate key assumptions. For example, the model requires that the dimensionless parameter B, defined here through normalization of Biot's equations, is much larger than one. This indicates that solid-fluid coupling forces are dominated by viscous rather than inertial effects. A tabulation of physical and kinematic variables for the rod-array experiments of Iverson and LaHusen and for various geologic phenomena shows that the model assumptions commonly are satisfied. A subsequent paper will describe model tests against experimental data. ?? 1993 International Association for Mathematical Geology.

  8. Random/aligned electrospun PCL/PCL-collagen nanofibrous membranes: comparison of neural differentiation of rat AdMSCs and BMSCs

    International Nuclear Information System (INIS)

    Çapkın, Merve; Gümüşderelioğlu, Menemşe; Çakmak, Soner; Kurt, Feyzan Özdal; Şen, B Hakan; Türk, B Tuğba; Deliloğlu-Gürhan, S İsmet

    2012-01-01

    In this study, the aligned (A) and randomly oriented (R) polycaprolactone (PCL-A and PCL-R) and PCL/collagen (PCL/Col-A and PCL/Col-R) nanofibers were electrospun onto smooth PCL membranes (PCLMs) prepared by solvent casting. In order to investigate the effects of chemical composition and nanotopography of fibrous surfaces on proliferation and on neural differentiation of mesenchymal stem cells (MSCs), adipose and bone marrow-derived rat MSCs (AdMSCs and BMSCs) were cultivated in suitable media i.e. inducing medium containing basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF), and cell maintenance medium (CMM). BMSCs adhered and proliferated on all nanofibrous membranes more efficiently than AdMSCs. PCL/Col-A was found as the most convenient surface supporting proliferation in both cell types. Immunofluorescence staining indicated that BMSCs and AdMSCs are prone for differentiation to oligodendrocytes more than they differentiate to other neuronal cell types. PCL-A nanofibrous membranes supported differentiation of MSCs to O4 + (an oligodendrocytes surface antigen) cells in both culture media. The intensity of immunoreactivity of O4 + cells differentiated from BMSCs on PCL-A was highest when compared with the other groups (p + cells. In conclusion, this study can be evaluated to establish the cell therapy strategies in neurodegenerative disorders, which are relevant to oligodendrocyte abstinence using BMSCs or AdMSCs on aligned nanofibrous membranes. (paper)

  9. SOX2 and OCT4 mRNA-expressing cells, detected by molecular beacons, localize to the center of neurospheres during differentiation.

    Directory of Open Access Journals (Sweden)

    Mirolyuba Ilieva

    Full Text Available Neurospheres are used as in vitro assay to measure the properties of neural stem cells. To investigate the molecular and phenotypic heterogeneity of neurospheres, molecular beacons (MBs targeted against the stem cell markers OCT4 and SOX2 were designed, and synthesized with a 2'-O-methyl RNA backbone. OCT4 and SOX2 MBs were transfected into human embryonic mesencephalon derived cells, which spontaneously form neurospheres when grown on poly-L-ornitine/fibronectin matrix and medium complemented with bFGF. OCT4 and SOX2 gene expression were tracked in individual cell using the MBs. Quantitative image analysis every day for seven days showed that the OCT4 and SOX2 mRNA-expressing cells clustered in the centre of the neurospheres cultured in differentiation medium. By contrast, cells at the periphery of the differentiating spheres developed neurite outgrowths and expressed the tyrosine hydroxylase protein, indicating terminal differentiation. Neurospheres cultured in growth medium contained OCT4 and SOX2-positive cells distributed throughout the entire sphere, and no differentiating neurones. Gene expression of SOX2 and OCT4 mRNA detected by MBs correlated well with gene and protein expression measured by qRT-PCR and immunostaining, respectively. These experimental data support the theoretical model that stem cells cluster in the centre of neurospheres, and demonstrate the use of MBs for the spatial localization of specific gene-expressing cells within heterogeneous cell populations.

  10. A novel chemical-defined medium with bFGF and N2B27 supplements supports undifferentiated growth in human embryonic stem cells

    International Nuclear Information System (INIS)

    Liu Yanxia; Song Zhihua; Zhao Yang; Qin Han; Cai Jun; Zhang Hong; Yu Tianxin; Jiang Siming; Wang Guangwen; Ding Mingxiao; Deng Hongkui

    2006-01-01

    Traditionally, undifferentiated human embryonic stem cells (hESCs) are maintained on mouse embryonic fibroblast (MEF) cells or on matrigel with an MEF-conditioned medium (CM), which hampers the clinical applications of hESCs due to the contamination by animal pathogens. Here we report a novel chemical-defined medium using DMEM/F12 supplemented with N2, B27, and basic fibroblast growth factor (bFGF) [termed NBF]. This medium can support prolonged self-renewal of hESCs. hESCs cultured in NBF maintain an undifferentiated state and normal karyotype, are able to form embryoid bodies in vitro, and differentiate into three germ layers and extraembryonic cells. Furthermore, we find that hESCs cultured in NBF possess a low apoptosis rate and a high proliferation rate compared with those cultured in MEF-CM. Our findings provide a novel, simplified chemical-defined culture medium suitable for further therapeutic applications and developmental studies of hESCs

  11. Sex Differences in Medium Spiny Neuron Excitability and Glutamatergic Synaptic Input: Heterogeneity Across Striatal Regions and Evidence for Estradiol-Dependent Sexual Differentiation

    Directory of Open Access Journals (Sweden)

    Jinyan Cao

    2018-04-01

    Full Text Available Steroid sex hormones and biological sex influence how the brain regulates motivated behavior, reward, and sensorimotor function in both normal and pathological contexts. Investigations into the underlying neural mechanisms have targeted the striatal brain regions, including the caudate–putamen, nucleus accumbens core (AcbC, and shell. These brain regions are of particular interest to neuroendocrinologists given that they express membrane-associated but not nuclear estrogen receptors, and also the well-established role of the sex steroid hormone 17β-estradiol (estradiol in modulating striatal dopamine systems. Indeed, output neurons of the striatum, the medium spiny neurons (MSNs, exhibit estradiol sensitivity and sex differences in electrophysiological properties. Here, we review sex differences in rat MSN glutamatergic synaptic input and intrinsic excitability across striatal regions, including evidence for estradiol-mediated sexual differentiation in the nucleus AcbC. In prepubertal animals, female MSNs in the caudate–putamen exhibit a greater intrinsic excitability relative to male MSNs, but no sex differences are detected in excitatory synaptic input. Alternatively, female MSNs in the nucleus AcbC exhibit increased excitatory synaptic input relative to male MSNs, but no sex differences in intrinsic excitability were detected. Increased excitatory synaptic input onto female MSNs in the nucleus AcbC is abolished after masculinizing estradiol or testosterone exposure during the neonatal critical period. No sex differences are detected in MSNs in prepubertal nucleus accumbens shell. Thus, despite possessing the same neuron type, striatal regions exhibit heterogeneity in sex differences in MSN electrophysiological properties, which likely contribute to the sex differences observed in striatal function.

  12. Collision density approach of radiation damage in a multispecies medium

    International Nuclear Information System (INIS)

    Lux, I.; Pazsit, I.

    1981-01-01

    Space-energy dependent forward type equations for the collision densities of energetic atoms in a multispecies semi-infinite homogeneous medium are formulated. Introduction of the one-dimensional isotropic forward-backward model of Fermi for the scattering and application of the Laplace transform with respect to the lethargy variable will lead to a linear differential equation system with constant coefficients. This equation system is solved for an arbitrary number of species and relations between the collision densities and defect distributions of the different species are given in the Kinchin-Pease model of radiation damage. The case of an alien particle incident on a two-component target is examined in some detail and the sputtering spectra for the three species are given numerically. (author)

  13. Collision density approach of radiation damage in a multispecies medium

    Energy Technology Data Exchange (ETDEWEB)

    Lux, I; Pazsit, I [Koezponti Elelmiszeripari Kutato Intezet, Budapest (Hungary)

    1981-01-01

    Space-energy dependent forward type equations for the collision densities of energetic atoms in a multispecies semi-infinite homogeneous medium are formulated. Introduction of the one-dimensional isotropic forward-backward model of Fermi for the scattering and application of the Laplace transform with respect to the lethargy variable will lead to a linear differential equation system with constant coefficients. This equation system is solved for an arbitrary number of species and relations between the collision densities and defect distributions of the different species are given in the Kinchin-Pease model of radiation damage. The case of an alien particle incident on a two-component target is examined in some detail and the sputtering spectra for the three species are given numerically.

  14. A simple and low cost dual-wavelength β-correction spectrophotometric determination and speciation of mercury(II) in water using chromogenic reagent 4-(2-thiazolylazo) resorcinol

    Science.gov (United States)

    Al-Bagawi, A. H.; Ahmad, W.; Saigl, Z. M.; Alwael, H.; Al-Harbi, E. A.; El-Shahawi, M. S.

    2017-12-01

    The most common problems in spectrophotometric determination of various complex species originate from the background spectral interference. Thus, the present study aimed to overcome the spectral matrix interference for the precise analysis and speciation of mercury(II) in water by dual-wavelength β-correction spectrophotometry using 4-(2-thiazolylazo) resorcinol (TAR) as chromogenic reagent. The principle was based on measuring the correct absorbance for the formed complex of mercury(II) ions with TAR reagent at 547 nm (lambda max). Under optimized conditions, a linear dynamic range of 0.1-2.0 μg mL- 1 with correlation coefficient (R2) of 0.997 were obtained with lower limits of detection (LOD) of 0.024 μg mL- 1 and limit of quantification (LOQ) of 0.081 μg mL- 1. The values of RSD and relative error (RE) obtained for β-correction method and single wavelength spectrophotometry were 1.3, 1.32% and 4.7, 5.9%, respectively. The method was validated in tap and sea water in terms of the data obtained from inductively coupled plasma-optical emission spectrometry (ICP-OES) using student's t and F tests. The developed methodology satisfactorily overcomes the spectral interference in trace determination and speciation of mercury(II) ions in water.

  15. Analysis of combined conduction and radiation heat transfer in presence of participating medium by the development of hybrid method

    International Nuclear Information System (INIS)

    Mahapatra, S.K.; Dandapat, B.K.; Sarkar, A.

    2006-01-01

    The current study addresses the mathematical modeling aspects of coupled conductive and radiative heat transfer in the presence of absorbing, emitting and isotropic scattering gray medium within two-dimensional square enclosure. A blended method where the concepts of modified differential approximation employed by combining discrete ordinate method and spherical harmonics method, has been developed for modeling the radiative transport equation. The gray participating medium is bounded by isothermal walls of two-dimensional enclosure which are considered to be opaque, diffuse and gray. The effect of various influencing parameters i.e., radiation-conduction parameter, surface emissivity, single scattering albedo and optical thickness has been illustrated. The adaptability of the present method has also been addressed

  16. SECTORAL STRUCTURE OF SMALL AND MEDIUM BUSINESS IN RUSSIA: STATISTICAL ANALYSIS

    Directory of Open Access Journals (Sweden)

    Yu. S. Pin’kovetskaya

    2018-01-01

    Full Text Available The research urgency is caused by the implementation of the Federal strategy of development of small and medium business for the period up to 2030.The purpose of the study: determination of the regularities characteristic of the sectoral structure of small and medium business. Objectives of the study: to establish the patterns and trends, characterizing the current structure of small and medium business by types of economic activity and the number of their employees, the definition of specific indicators of the number of employees per enterprise, as well as estimation of the distribution of these parameters on the aggregates of entrepreneurial structures in the regions. The results of continuous monitoring activities of small and medium business in 2015 were used as input data. The study was based on the comparison of indicators for the business sector and the full circle of legal entities and individual entrepreneurs.Research methodology is based on the consideration of sets of entities generated by the sectoral and territorial characteristics. Modeling of the values’ differentiation of the number of workers per enterprise based on the development of the density function of the normal distribution. The quality of the developed models was verified with Kolmogorov-Smirnov, Pearson and Shapiro-Wilk. According to the results of modeling, the mean values of the number of workers on six main activities are set, as well as the intervals of changes of these parameters on the aggregates of small and medium enterprises, located in most regions of the country.The obtained results have some theoretical significance, in particular, when conducting research, related to the justification of the number of enterprises and number of employees, the formation of measures for the increase of efficiency of activity of the business sector. Given in the article, density function of the normal distribution can be used to establish the plans and programs of

  17. In Vivo Differentiation of Complementary Contrast Media at Dual-Energy CT

    Science.gov (United States)

    Mongan, John; Rathnayake, Samira; Fu, Yanjun; Wang, Runtang; Jones, Ella F.; Gao, Dong-Wei

    2012-01-01

    Purpose: To evaluate the feasibility of using a commercially available clinical dual-energy computed tomographic (CT) scanner to differentiate the in vivo enhancement due to two simultaneously administered contrast media with complementary x-ray attenuation ratios. Materials and Methods: Approval from the institutional animal care and use committee was obtained, and National Institutes of Health guidelines for the care and use of laboratory animals were observed. Dual-energy CT was performed in a set of iodine and tungsten solution phantoms and in a rabbit in which iodinated intravenous and bismuth subsalicylate oral contrast media were administered. In addition, a second rabbit was studied after intravenous administration of iodinated and tungsten cluster contrast media. Images were processed to produce virtual monochromatic images that simulated the appearance of conventional single-energy scans, as well as material decomposition images that separate the attenuation due to each contrast medium. Results: Clear separation of each of the contrast media pairs was seen in the phantom and in both in vivo animal models. Separation of bowel lumen from vascular contrast medium allowed visualization of bowel wall enhancement that was obscured by intraluminal bowel contrast medium on conventional CT scans. Separation of two vascular contrast media in different vascular phases enabled acquisition of a perfectly coregistered CT angiogram and venous phase–enhanced CT scan simultaneously in a single examination. Conclusion: Commercially available clinical dual-energy CT scanners can help differentiate the enhancement of selected pairs of complementary contrast media in vivo. © RSNA, 2012 PMID:22778447

  18. Collision density approach of radiation damage in a multi-species medium

    International Nuclear Information System (INIS)

    Lux, I.; Pazsit, I.

    1981-05-01

    Space-energy dependent foward type equations for the collision densities of energetic atoms in multi-species semi-infinite homogeneous medium are formulated. The introduction of the one-dimensional isotropic forward-backward model of Fermi for the scattering and application of the Laplace transformation with respect to the lethargy variable leads to a linear differential equation system with constant coefficients. This equation system is solved for an arbitrary number of species and relations between the collision densities and defect distributions of the different species are given in the Kinchin Pease model. The case of an alien particle incident on a two-component target is examined in some detail and the sputtering spectra are given numerically. (author)

  19. Divalent Metal Ions Induced Osteogenic Differentiation of MC3T3E1

    Science.gov (United States)

    Wang, Guoshou; Su, Wenta; Chen, Pohung; Huang, Teyang

    2017-12-01

    Biomaterial scaffolds blended with biochemical signal molecules with adequate osteoinductive and osteoconductive properties have attracted significant interest in bone tissue engineering regeneration. The divalent metal ions can gradually release from the scaffold into the culture medium and then induced osteoblastic differentiation of MC3T3E1. These MC3T3E1 cells expressed high activity of alkaline phosphatase, bone-related gene expression of collagen type I, Runx2, osteopontin, osteocalcin, and significantly enhanced deposited minerals on scaffold after 21 days of culture. This experiment provided a useful inducer for osteogenic differentiation in bone repair.

  20. Utility of chromogenic in situ hybridization (CISH) for detection of EGFR amplification in glioblastoma: comparison with fluorescence in situ hybridization (FISH).

    Science.gov (United States)

    Fischer, Ingeborg; de la Cruz, Clarissa; Rivera, Andreana L; Aldape, Kenneth

    2008-12-01

    In this study, we test the reliability of chromogenic in situ hybridization (CISH) for the detection of epidermal growth factor receptor (EGFR) gene amplification in glioblastoma. Earlier reports have described EGFR CISH in glioblastoma multiforme, but a comparison of CISH with a "gold standard" testing method, such as fluorescence in situ hybridization (FISH), has not been described. Therapies targeting the EGFR-signaling pathway might increase the importance of assessment of EGFR-amplification status. CISH is a potential alternative to FISH as a testing method. To test its reliability, EGFR-amplification status by CISH was assessed in 89 cases of glioblastoma and compared with FISH results, and correlated with the protein expression using immunohistochemistry (IHC) for EGFR. FISH was scored as being EGFR-amplified in 47/89 tumors, CISH as being amplified in 43/89 tumors. The CISH and FISH results were in agreement in 83/89 cases (93%). Four glioblastomas were scored as being amplified by FISH, but not by CISH; whereas amplification was detected in 2 tumors by CISH that were not amplified using FISH. Forty-eight of the 89 cases were positive for EGFR expression by IHC. EGFR amplification was highly correlated with protein expression by IHC, as 40/48 (83%) EGFR IHC-positive cases were found to be EGFR-amplified. The high concordance of CISH and FISH for the assessment of EGFR gene-amplification status indicates that CISH is a viable alternative to FISH for the detection of EGFR gene amplification in glioblastoma. Detectable EGFR expression by IHC can occur in the absence of gene amplification, but is uncommon.

  1. Differentiation of car rental services in the Croatian market

    Directory of Open Access Journals (Sweden)

    Karolina Karlo Marijanović

    2010-06-01

    Full Text Available A number of service companies focus on service differentiation nowadays. The companies that manage to deliver superior service are winners and may serve as an example of how the growth of services has changed business. The first part of the paper presents an analysis of the Croatian car rental services market. The theoretical part of the paper provides an insight into the characteristics of services in general, followed by an overview of the characteristics and features of the companies operating in the local market. The second part of the paper deals with the work of Porter and his three generic strategies, and with his differentiation strategy in particular. Subsequently, all possibilities of differentiation strategies that are appropriate to the car rental services are analyzed. Beside global recession, the Croatian market is currently also struggling with huge competition and companies cannot generate profit without continuous differentiation of all elements of service. In the third chapter the following hypothesis is set: By a continuous differentiation of all elements of the services marketing mix small businesses can compete with large international companies. Acceptance or rejection of the hypothesis is shown by the research conducted among users of the specific service. Its results fully confirm the set hypothesis. The fact that, despite global recession, the majority of respondents believes the survival of small- and medium-sized companies which continuously differentiate their business activities to be unquestionable is indeed encouraging.

  2. Longevity of U cells of differentiated yeast colonies grown on respiratory medium depends on active glycolysis

    Czech Academy of Sciences Publication Activity Database

    Čáp, M.; Váchová, Libuše; Palková, Z.

    2015-01-01

    Roč. 14, č. 21 (2015), s. 3488-3497 ISSN 1538-4101 R&D Projects: GA ČR(CZ) GA15-08225S; GA MŠk(CZ) ED1.1.00/02.0109 Institutional support: RVO:61388971 Keywords : enzymatic assays * fermentation * metabolic differentiation Subject RIV: EE - Microbiology, Virology Impact factor: 3.952, year: 2015

  3. A frame work for heat generation/absorption and modified homogeneous–heterogeneous reaction in flow based on non-Darcy–Forchheimer medium

    Directory of Open Access Journals (Sweden)

    Tasawar Hayat

    2018-04-01

    Full Text Available The present work aims to report the consequences of Darcy–Forchheimer medium in flow of Cross fluid model toward a stretched surface. Flow in porous space is categorized by Darcy–Forchheimer medium. Further heat transfer characteristics are examined via thermal radiation and heat generation/absorption. Transformation procedure is used. The arising system of nonlinear ordinary differential equations is solved numerically by means of shooting method. The effects of different flow variables on velocity, temperature, concentration, skin friction, and heat transfer rate are discussed. The obtained outcomes show that velocity was enhanced with the increase in the Weissenberg number but decays with increase in the porosity parameter and Hartman number. Temperature field is boosted by thermal radiation and heat generation; however, it decays with the increase in the Prandtl number. Keywords: Cross Fluid, Heat Generation/Absorption, Homogeneous–Heterogeneous Reactions, Non-Darcy–Forchheimer Medium, Thermal Radiation

  4. Current View on Osteogenic Differentiation Potential of Mesenchymal Stromal Cells Derived from Placental Tissues.

    Science.gov (United States)

    Kmiecik, Gabriela; Spoldi, Valentina; Silini, Antonietta; Parolini, Ornella

    2015-08-01

    Mesenchymal stromal cells (MSC) isolated from human term placental tissues possess unique characteristics, including their peculiar immunomodulatory properties and their multilineage differentiation potential. The osteogenic differentiation capacity of placental MSC has been widely disputed, and continues to be an issue of debate. This review will briefly discuss the different MSC populations which can be obtained from different regions of human term placenta, along with their unique properties, focusing specifically on their osteogenic differentiation potential. We will present the strategies used to enhance osteogenic differentiation potential in vitro, such as through the selection of subpopulations more prone to differentiate, the modification of the components of osteo-inductive medium, and even mechanical stimulation. Accordingly, the applications of three-dimensional environments in vitro and in vivo, such as non-synthetic, polymer-based, and ceramic scaffolds, will also be discussed, along with results obtained from pre-clinical studies of placental MSC for the regeneration of bone defects and treatment of bone-related diseases.

  5. Functional differential equations for the q-Fourier transform of q-Gaussians

    International Nuclear Information System (INIS)

    Umarov, S; Queiros, S M Duarte

    2010-01-01

    In this paper the question 'is the q-Fourier transform of a q-Gaussian a q'-Gaussian (with some q') up to a constant factor?' is studied for the whole range of q in (- infty, 3). This question is connected with applicability of the q-Fourier transform in the study of limit processes in nonextensive statistical mechanics. Using the functional differential equation approach we prove that the answer is affirmative if and only if 1 ≤ q < 3, excluding two particular cases of q < 1, namely q=1/2 and q=2/3. Complementarily, we discuss some applications of the q-Fourier transform to nonlinear partial differential equations such as the porous medium equation.

  6. Functional differential equations for the q-Fourier transform of q-Gaussians

    Energy Technology Data Exchange (ETDEWEB)

    Umarov, S [Department of Mathematics, Tufts University, Medford, MA (United States); Queiros, S M Duarte, E-mail: sdqueiro@gmail.co [Unilever R and D Port Sunlight, Quarry Road East, Wirral, CH63 3JW (United Kingdom)

    2010-02-05

    In this paper the question 'is the q-Fourier transform of a q-Gaussian a q'-Gaussian (with some q') up to a constant factor?' is studied for the whole range of q in (- infty, 3). This question is connected with applicability of the q-Fourier transform in the study of limit processes in nonextensive statistical mechanics. Using the functional differential equation approach we prove that the answer is affirmative if and only if 1 <= q < 3, excluding two particular cases of q < 1, namely q=1/2 and q=2/3. Complementarily, we discuss some applications of the q-Fourier transform to nonlinear partial differential equations such as the porous medium equation.

  7. Overexpression of α-catenin increases osteoblastic differentiation in mouse mesenchymal C3H10T1/2 cells

    International Nuclear Information System (INIS)

    Kim, Dohee; Yang, Jae-Yeon; Shin, Chan Soo

    2009-01-01

    α- and β-Catenin link cadherins to the actin-based cytoskeleton at adherens junctions and regulate cell-cell adhesion. Although roles of cadherins and canonical Wnt-/β-catenin-signaling in osteoblastic differentiation have been extensively studied, the role of α-catenin is not known. Murine embryonic mesenchymal stem cells, C3H10T1/2 cells, were transduced with retrovirus encoding α-catenin (MSCV-α-catenin-HA-GFP). In the presence of Wnt-3A conditioned medium or osteogenic medium (β-glycerol phosphate and ascorbic acid), cells overexpressing α-catenin showed enhanced osteoblastic differentiation as measured by alkaline phosphatase (ALP) staining and ALP activity assay compared to cells transduced with empty virus (MSCV-GFP). In addition, mRNA expression of osteocalcin and Runx2 was significantly increased compared to control. Cell aggregation assay revealed that α-catenin overexpression has significantly increased cell-cell aggregation. However, cellular β-catenin levels (total, cytoplasmic-nuclear ratio) and β-catenin-TCF/LEF transcriptional activity did not change by overexpression of α-catenin. Knock-down of α-catenin using siRNA decreased osteoblastic differentiation as measured by ALP assay. These results suggest that α-catenin overexpression increases osteoblastic differentiation by increasing cell-cell adhesion rather than Wnt-/β-catenin-signaling.

  8. 27 CFR 19.914 - Medium plants.

    Science.gov (United States)

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Medium plants. 19.914... OF THE TREASURY LIQUORS DISTILLED SPIRITS PLANTS Distilled Spirits For Fuel Use Permits § 19.914 Medium plants. Any person wishing to establish a medium plant shall make application for and obtain in...

  9. Different culture media affect growth characteristics, surface marker distribution and chondrogenic differentiation of human bone marrow-derived mesenchymal stromal cells.

    Science.gov (United States)

    Hagmann, Sebastien; Moradi, Babak; Frank, Sebastian; Dreher, Thomas; Kämmerer, Peer Wolfgang; Richter, Wiltrud; Gotterbarm, Tobias

    2013-07-30

    Bone marrow-derived mesenchymal stromal cells (BM-MSCs) play an important role in modern tissue engineering, while distinct variations of culture media compositions and supplements have been reported. Because MSCs are heterogeneous regarding their regenerative potential and their surface markers, these parameters were compared in four widely used culture media compositions. MSCs were isolated from bone marrow and expanded in four established cell culture media. MSC yield/1000 MNCs, passage time and growth index were observed. In P4, typical MSC surface markers were analysed by fluorescence cytometry. Additionally, chondrogenic, adipogenic and osteogenic differentiation potential were evaluated. Growth index and P0 cell yield varied importantly between the media. The different expansion media had a significant influence on the expression of CD10, CD90, CD105, CD140b CD146 and STRO-1. While no significant differences were observed regarding osteogenic and adipogenic differentiation, chondrogenic differentiation was superior in medium A as reflected by GAG/DNA content. The choice of expansion medium can have a significant influence on growth, differentiation potential and surface marker expression of mesenchymal stromal cells, which is of fundamental importance for tissue engineering procedures.

  10. S36. DIFFERENTIAL ENCODING OF SENSITIZATION AND CROSS SENSITIZATION TO PSYCHOSTIMULANTS AND ANTIPSYCHOTICS IN NUCLEUS ACCUMBENS D1- AND D2- RECEPTOR EXPRESSING MEDIUM SPINY NEURONS

    Science.gov (United States)

    Amato, Davide; Heinsbroek, Jasper; Kalivas, Peter W

    2018-01-01

    Abstract Background Nearly half of all individuals diagnosed with schizophrenia abuse addictive substances such as cocaine. Currently, the neurobiological mechanisms in patients with schizophrenia that lead to cocaine abuse are unknown. A possible explanation for the co-morbidity between schizophrenia and addiction is that the rewarding properties of cocaine reverse the diminished motivational drive caused by chronic antipsychotic regimen. Moreover, chronic antipsychotic treatment can sensitize and amplify cocaine rewarding effects and exacerbate psychoses. Methods The rewarding properties of cocaine are attributed to the differential effects of dopamine on D1 and D2 receptor-expressing medium spiny neurons (MSNs) in the nucleus accumbens (NAc). Using in vivo Ca2+ miniature microscopic imaging, we characterize the role of D1 and D2 MSN in mono- and a cross- sensitization paradigms. D1- and D2-Cre mice were injected with a Cre dependent calcium indicator (gCaMP6f) and implanted with a gradient index (GRIN) lens above the nucleus accumbens and calcium activity was recorded using a head mounted miniature microscope. Cocaine sensitization was measured after a classic repeated cocaine regiment and antipsychotic and psychostimulant cross-sensitization was measured by a single cocaine injection after chronic pre-treatment with haloperidol. Results We found that both D1-MSN and D2-MSN populations are modulated by initial cocaine experience and further modulated during the expression of cocaine sensitization. A subpopulation of D1-MSN displayed initial activation, but reduced activity during the expression of sensitization. By contrast, the majority of D2-MSNs were suppressed by initial cocaine experience, but became active during the expression of sensitization. Furthermore, activity of D1- and D2-MSNs bidirectionally related with the observed behavioral responses to cocaine. Cross-sensitization following haloperidol treatment led to increased behavioral responses to

  11. Isolation of Cronobacter spp. (formerly Enterobacter sakazakii) from infant food, herbs and environmental samples and the subsequent identification and confirmation of the isolates using biochemical, chromogenic assays, PCR and 16S rRNA sequencing.

    Science.gov (United States)

    Jaradat, Ziad W; Ababneh, Qotaiba O; Saadoun, Ismail M; Samara, Nawal A; Rashdan, Abrar M

    2009-10-27

    Cronobacter spp. (formerly Enterobacter sakazakii), are a group of Gram-negative pathogens that have been implicated as causative agents of meningitis and necrotizing enterocolitis in infants. The pathogens are linked to infant formula; however, they have also been isolated from a wide range of foods and environmental samples. In this study, 233 samples of food, infant formula and environment were screened for the presence of Cronobacter spp. in an attempt to find its source. Twenty nine strains were isolated from samples of spices, herbs, infant foods, and dust obtained from household vacuum cleaners. Among the 76 samples of infant food, infant formula, milk powder and non-milk dairy products tested, only one sample of infant food contained Cronobacter spp. (1.4%). The other Cronobacter spp. isolates recovered include two from household vacuum dust, and 26 from 67 samples of herbs and spices. Among the food categories analyzed, herbs and spices harbored the highest number of isolates, indicating plants as a possible reservoir of this pathogen. Initial screening with API 20E test strips yielded 42 presumptive isolates. Further characterization using 3 chromogenic media (alpha-MUG, DFI and EsPM) and 8 sets of PCR primers detecting ITS (internal transcribed spacer sequences), 16S rRNA, zpx, gluA, gluB, OmpA genes followed by nucleotide sequencing of some PCR amplicons did not confirm the identity of all the isolates as none of the methods proved to be free of both false positives or false negatives. The final confirmation step was done by 16S rRNA sequence analysis identifying only 29 of the 42 isolates as Cronobacter spp. Our studies showed that Cronobacter spp. are highly diverse and share many phenotypic traits with other Enterobacteriaceae members highlighting the need to use several methods to confirm the identity of this pathogen. None of the biochemical, chromogenic or PCR primers proved to be a reliable method for confirmation of the identity of the isolates

  12. Isolation of Cronobacter spp. (formerly Enterobacter sakazakii from infant food, herbs and environmental samples and the subsequent identification and confirmation of the isolates using biochemical, chromogenic assays, PCR and 16S rRNA sequencing

    Directory of Open Access Journals (Sweden)

    Samara Nawal A

    2009-10-01

    Full Text Available Abstract Background Cronobacter spp. (formerly Enterobacter sakazakii, are a group of Gram-negative pathogens that have been implicated as causative agents of meningitis and necrotizing enterocolitis in infants. The pathogens are linked to infant formula; however, they have also been isolated from a wide range of foods and environmental samples. Results In this study, 233 samples of food, infant formula and environment were screened for the presence of Cronobacter spp. in an attempt to find its source. Twenty nine strains were isolated from samples of spices, herbs, infant foods, and dust obtained from household vacuum cleaners. Among the 76 samples of infant food, infant formula, milk powder and non-milk dairy products tested, only one sample of infant food contained Cronobacter spp. (1.4%. The other Cronobacter spp. isolates recovered include two from household vacuum dust, and 26 from 67 samples of herbs and spices. Among the food categories analyzed, herbs and spices harbored the highest number of isolates, indicating plants as a possible reservoir of this pathogen. Initial screening with API 20E test strips yielded 42 presumptive isolates. Further characterization using 3 chromogenic media (α-MUG, DFI and EsPM and 8 sets of PCR primers detecting ITS (internal transcribed spacer sequences, 16S rRNA, zpx, gluA, gluB, OmpA genes followed by nucleotide sequencing of some PCR amplicons did not confirm the identity of all the isolates as none of the methods proved to be free of both false positives or false negatives. The final confirmation step was done by 16S rRNA sequence analysis identifying only 29 of the 42 isolates as Cronobacter spp. Conclusion Our studies showed that Cronobacter spp. are highly diverse and share many phenotypic traits with other Enterobacteriaceae members highlighting the need to use several methods to confirm the identity of this pathogen. None of the biochemical, chromogenic or PCR primers proved to be a reliable

  13. Evaluation of Insulin Medium or Chondrogenic Medium on Proliferation and Chondrogenesis of ATDC5 Cells

    OpenAIRE

    Yao, Yongchang; Zhai, Zhichen; Wang, Yingjun

    2014-01-01

    Background. The ATDC5 cell line is regarded as an excellent cell model for chondrogenesis. In most studies with ATDC5 cells, insulin medium (IM) was used to induce chondrogenesis while chondrogenic medium (CM), which was usually applied in chondrogenesis of mesenchymal stem cells (MSCs), was rarely used for ATDC5 cells. This study was mainly designed to investigate the effect of IM, CM, and growth medium (GM) on chondrogenesis of ATDC5 cells. Methods. ATDC5 cells were, respectively, cultured ...

  14. On the viscous dissipation modeling of thermal fluid flow in a porous medium

    KAUST Repository

    Salama, Amgad

    2011-02-24

    The problem of viscous dissipation and thermal dispersion in saturated porous medium is numerically investigated for the case of non-Darcy flow regime. The fluid is induced to flow upward by natural convection as a result of a semi-infinite vertical wall that is immersed in the porous medium and is kept at constant higher temperature. The boundary layer approximations were used to simplify the set of the governing, nonlinear partial differential equations, which were then non-dimensionalized and solved using the finite elements method. The results for the details of the governing parameters are presented and investigated. It is found that the irreversible process of transforming the kinetic energy of the moving fluid to heat energy via the viscosity of the moving fluid (i.e.; viscous dissipation) resulted in insignificant generation of heat for the range of parameters considered in this study. On the other hand, thermal dispersion has shown to disperse heat energy normal to the wall more effectively compared with the normal diffusion mechanism. © 2011 Springer-Verlag.

  15. Differential invariants in nonclassical models of hydrodynamics

    Science.gov (United States)

    Bublik, Vasily V.

    2017-10-01

    In this paper, differential invariants are used to construct solutions for equations of the dynamics of a viscous heat-conducting gas and the dynamics of a viscous incompressible fluid modified by nanopowder inoculators. To describe the dynamics of a viscous heat-conducting gas, we use the complete system of Navier—Stokes equations with allowance for heat fluxes. Mathematical description of the dynamics of liquid metals under high-energy external influences (laser radiation or plasma flow) includes, in addition to the Navier—Stokes system of an incompressible viscous fluid, also heat fluxes and processes of nonequilibrium crystallization of a deformable fluid. Differentially invariant solutions are a generalization of partially invariant solutions, and their active study for various models of continuous medium mechanics is just beginning. Differentially invariant solutions can also be considered as solutions with differential constraints; therefore, when developing them, the approaches and methods developed by the science schools of academicians N. N. Yanenko and A. F. Sidorov will be actively used. In the construction of partially invariant and differentially invariant solutions, there are overdetermined systems of differential equations that require a compatibility analysis. The algorithms for reducing such systems to involution in a finite number of steps are described by Cartan, Finikov, Kuranishi, and other authors. However, the difficultly foreseeable volume of intermediate calculations complicates their practical application. Therefore, the methods of computer algebra are actively used here, which largely helps in solving this difficult problem. It is proposed to use the constructed exact solutions as tests for formulas, algorithms and their software implementations when developing and creating numerical methods and computational program complexes. This combination of effective numerical methods, capable of solving a wide class of problems, with

  16. Hospital morbidity in a medium-sized city: differentials between men and women

    Directory of Open Access Journals (Sweden)

    Guilherme Oliveira de Arruda

    2014-01-01

    Full Text Available OBJECTIVE: characterize the hospital morbidity of adults living in the city of Maringá, PR, Brazil, between 2000 and 2011, focusing on the differential between men and women. METHOD: this descriptive study was developed based on data from the Hospital Information System of the Unified Health System in order to investigate the association between groups of hospitalization causes and the average length of hospitalization per gender, in three-year periods. RESULTS: the main groups of hospitalization causes for men were: mental disorders, lesions and circulatory diseases; and, among women: tumors, circulatory and genitourinary diseases. Mental disorders and lesions, tumors, circulatory and genitourinary diseases were significantly associated with the female and male genders across the study period. Although not significant, the mean length of hospitalization dropped across the four three-year periods, and only showed a significant difference between men and women in the second triennium. CONCLUSION: differences in the hospital morbidity profile between men and women underline the need for specific health and nursing actions, especially in primary health care, with a view to reducing hospitalizations due to the main groups of causes in the city.

  17. Pions in the nuclear medium

    International Nuclear Information System (INIS)

    Chanfray, G.

    1996-07-01

    We discuss various aspects of pion physics in the nuclear medium. We first study s-wave pion-nucleus interaction in connection with chiral symmetry restoration and quark condensate in the nuclear medium. We then address the question of p-wave pion-nucleus interaction and collective pionic modes in nuclei and draw the consequences for in medium ππ correlations especially in the scalar-isoscalar channel. We finally discuss the modification of the rho meson mass spectrum at finite density and/or temperature in connection with relativistic heavy ion collisions

  18. Optical recording medium

    International Nuclear Information System (INIS)

    Andriech, A.; Bivol, V.; Tridukh, G.; Tsiuleanu, D.

    2002-01-01

    The invention relates of the micro- and optoelectronics, computer engineering ,in particular, to tjhe optical information media and may be used in hilography. Summary of the invention consists in that the optical image recording medium, containing a dielectric substrates, onto one surface of which there are placed in series a transparent electricity conducting layer, a photo sensitive recording layer of chalcogenic glass and a thin film electrode of aluminium, is provided with an optically transparent protective layer, applied into the thin film electrode. The result of the invention consists in excluding the dependence of chemical processes course into the medium upon environmental conditions

  19. Irreversibility analysis of hydromagnetic flow of couple stress fluid with radiative heat in a channel filled with a porous medium

    Directory of Open Access Journals (Sweden)

    A.S. Eegunjobi

    Full Text Available Numerical analysis of the intrinsic irreversibility of a mixed convection hydromagnetic flow of an electrically conducting couple stress fluid through upright channel filled with a saturated porous medium and radiative heat transfer was carried out. The thermodynamics first and second laws were employed to examine the problem. We obtained the dimensionless nonlinear differential equations and solves numerically with shooting procedure joined with a fourth order Runge-Kutta-Fehlberg integration scheme. The temperature and velocity obtained, used to analyse the entropy generation rate together with some various physical parameters of the flow. Our results are presented graphically and talk over. Keywords: MHD channel flow, Couple stress fluid, Porous medium, Thermal radiation, Entropy generation, Injection/suction

  20. Erythroid differentiation and commitment in rat erythroleukemia cells with hypertonic culture conditions.

    OpenAIRE

    Yamaguchi, Y; Kluge, N; Ostertag, W; Furusawa, M

    1981-01-01

    Cell cultures of 7,12-dimethylbenz[a]anthracene-induced rat erythroleukemia can be stimulated to synthesize hemoglobin when cultured in hypertonic media. During hypertonic treatment the intracellular osmotic conditions immediately readjust to those of the extracellular medium. None of the Friend virus-induced mouse erythroleukemia cell lines was inducible for differentiation with the same hypertonic culture conditions used for rat cells. Earliest commitment to erythroid terminal differentiati...

  1. Chromogenic in situ hybridization (CISH) to detect HER2 gene amplification in breast and gastric cancer: comparison with immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH).

    Science.gov (United States)

    Kiyose, Shinichiro; Igarashi, Hisaki; Nagura, Kiyoko; Kamo, Takaharu; Kawane, Kazunori; Mori, Hiroki; Ozawa, Takachika; Maeda, Matsuyoshi; Konno, Keisuke; Hoshino, Hideaki; Konno, Hiroyuki; Ogura, Hiroyuki; Shinmura, Kazuya; Hattori, Naohiko; Sugimura, Haruhiko

    2012-11-01

    The chromogenic in situ hybridization (CISH) assay, designed to detect the amplification of the HER2 gene in formalin-fixed, paraffin-embedded (FFPE) breast cancer (BC) and gastric cancer (GC) tissue specimens, was evaluated in 125 FFPE BC cases and 198 FFPE GC cases for which the HER2 status had been predetermined using immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH). In the 125 BC cases and the 198 gastric cases, we found a very good concordance (98.4% and 99.0%, respectively) between CISH and FISH. In particular, we evaluated the polysomy cases, as these cases often have ambiguous treatment options in clinical practice. The polysomy of chromosome 17 was defined as the presence of three or more CEP17 signals in at least 10% of the tumor cells. In the 50 BC cases and 54 GC cases displaying chromosome 17 polysomy, the concordance between FISH and CISH was 98.0% and 98.1%, respectively. These results indicate that CISH could provide an accurate and practical alternative to FISH for the clinical diagnosis of HER2 gene amplification in FFPE BC and FFPE GC samples. © 2012 The Authors. Pathology International © 2012 Japanese Society of Pathology and Wiley Publishing Asia Pty Ltd.

  2. Glycometabolic reprogramming associated with the initiation of human dental pulp stem cell differentiation.

    Science.gov (United States)

    Wang, Linyan; Cheng, Li; Wang, Huning; Pan, Hongying; Yang, Hui; Shao, Meiying; Hu, Tao

    2016-03-01

    Glycometabolism, particularly mitochondrial oxidative phosphorylation (OXPHOS) and glycolysis, plays a central role in cell life activities. Glycometabolism can be reprogrammed to maintain the stemness or to induce the differentiation of stem cells, thereby regulating tissue repair and regeneration. However, research on the glycometabolism of human dental pulp stem cells (hDPSCs) remains scarce. Here, we investigated the relationship between glycometabolic reprogramming and initiation of hDPSC differentiation. We found the differentiation of hDPSCs commenced on day 3 when cells were cultured in mineralized medium. When cell differentiation commenced, mitochondria became elongated with well-developed cristae, and the oxygen consumption rate of mitochondria was enhanced, manifested as an increase in basal respiration, mitochondrial ATP production, and maximal respiration. Interestingly, glycolytic enzyme activities, glycolysis capacity, and glycolysis reserve were also upregulated at this time to match the powerful bioenergetic demands. More importantly, hDPSCs derived from different donors or cultured in various oxygen environments showed similar glycometabolic changes when they began to differentiate. Thus, glycometabolic reprogramming accompanies initiation of hDPSC differentiation and could potentially play a role in the regulation of dental pulp repair. © 2015 International Federation for Cell Biology.

  3. Collaborative Manufacturing for Small-Medium Enterprises

    Science.gov (United States)

    Irianto, D.

    2016-02-01

    Manufacturing systems involve decisions concerning production processes, capacity, planning, and control. In a MTO manufacturing systems, strategic decisions concerning fulfilment of customer requirement, manufacturing cost, and due date of delivery are the most important. In order to accelerate the decision making process, research on decision making structure when receiving order and sequencing activities under limited capacity is required. An effective decision making process is typically required by small-medium components and tools maker as supporting industries to large industries. On one side, metal small-medium enterprises are expected to produce parts, components or tools (i.e. jigs, fixture, mold, and dies) with high precision, low cost, and exact delivery time. On the other side, a metal small- medium enterprise may have weak bargaining position due to aspects such as low production capacity, limited budget for material procurement, and limited high precision machine and equipment. Instead of receiving order exclusively, a small-medium enterprise can collaborate with other small-medium enterprise in order to fulfill requirements high quality, low manufacturing cost, and just in time delivery. Small-medium enterprises can share their best capabilities to form effective supporting industries. Independent body such as community service at university can take a role as a collaboration manager. The Laboratory of Production Systems at Bandung Institute of Technology has implemented shared manufacturing systems for small-medium enterprise collaboration.

  4. Nonlinear radiative peristaltic flow of hydromagnetic fluid through porous medium

    Science.gov (United States)

    Hussain, Q.; Latif, T.; Alvi, N.; Asghar, S.

    2018-06-01

    The radiative heat and mass transfer in wall induced flow of hydromagnetic fluid through porous medium in an asymmetric channel is analyzed. The fluid viscosity is considered temperature dependent. In the theory of peristalsis, the radiation effects are either ignored or taken as linear approximation of radiative heat flux. Such approximation is only possible when there is sufficiently small temperature differences in the flow field; however, nonlinear radiation effects are valid for large temperature differences as well (the new feature added in the present study). Mathematical modeling of the problems include the complicated system of highly nonlinear differential equations. Semi-analytical solutions are established in the wave reference frame. Results are displayed graphically and discussed in detail for the variation of various physical parameters with the special attention to viscosity, radiation, and temperature ratio parameters.

  5. Properties of the nuclear medium

    International Nuclear Information System (INIS)

    Baldo, M; Burgio, G F

    2012-01-01

    We review our knowledge on the properties of the nuclear medium that have been studied, over many years, on the basis of many-body theory, laboratory experiments and astrophysical observations. Throughout the presentation particular emphasis is placed on the possible relationship and links between the nuclear medium and the structure of nuclei, including the limitations of such an approach. First we consider the realm of phenomenological laboratory data and astrophysical observations and the hints they can give on the characteristics that the nuclear medium should possess. The analysis is based on phenomenological models, that however have a strong basis on physical intuition and an impressive success. More microscopic models are also considered, and it is shown that they are able to give invaluable information on the nuclear medium, in particular on its equation of state. The interplay between laboratory experiments and astrophysical observations is particularly stressed, and it is shown how their complementarity enormously enriches our insights into the structure of the nuclear medium. We then introduce the nucleon–nucleon interaction and the microscopic many-body theory of nuclear matter, with a critical discussion about the different approaches and their results. The Landau–Fermi liquid theory is introduced and briefly discussed, and it is shown how fruitful it can be in discussing the macroscopic and low-energy properties of the nuclear medium. As an illustrative example, we discuss neutron matter at very low density, and it is shown how it can be treated within the many-body theory. The general bulk properties of the nuclear medium are reviewed to indicate at which stage of our knowledge we stand, taking into account the most recent developments both in theory and experiments. A section is dedicated to the pairing problem. The connection with nuclear structure is then discussed, on the basis of the energy density functional method. The possibility of

  6. Effect of bionic electrical stimulation on the differentiation of embryonic stem cells into cardiomyocytes in the presence myocardial cells in vitro

    Directory of Open Access Journals (Sweden)

    Li-na ZHENG

    2011-08-01

    Full Text Available Objective To investigate the effects of electrical stimulation on the differentiation of embryonic stem cells(ESCs into cardiomyocytes in the presence of myocardial cells in vitro.Methods ESCs and neonate rat cardiomyocytes were isolated and cultured.These cells of primary culture were divided into 5 groups according to whether or not electric stimulation was given and the presence of cardiomyocytes: control group,stimulation group,cardiomyocytes group,stimulation+ cardiomyocyte conditioned medium group,and stimulation+cardiomyocytes group.Expression of troponin T(cTnT in the differentiated cells from ESCs was examined by immunofluoresence on the 5th,7th and 14th day.Results In the group co-cultured with myocardial cell and electrical stimulation,the differentiating ratio of cardiomyocytes derived from ESCs and expressing cTnT was 40.00%±2.39%,and it was higher than that in control group(2.00%±1.60%,stimulation group(3.00%±2.00%,cardiomyocytes group(28.70%±4.06%,stimulation+cardiomyocyte conditioned medium group(17.10%±2.23%,P < 0.05.Conclusion Bionic electric stimulation promotes the differentiation of ESCs into cardiomyocyte in a microenvironment consisting of myocardial cells.

  7. Forced convection on a heated horizontal flat plate with finite thermal conductivity in a non-Darcian porous medium

    Energy Technology Data Exchange (ETDEWEB)

    Luna, N. [Direccion de Operacion Petrolera, Direccion General de Exploracion y Explotacion de Hidrocarburos, Secretaria de Energia, 03100 Mexico DF (Mexico); Mendez, F. [Facultad de Ingenieria, UNAM, 04510 Mexico DF (Mexico)

    2005-07-01

    The steady-state analysis of conjugated heat transfer process for the hydrodynamically developed forced convection flow on a heated flat plate embedded in a porous medium is studied. The governing equations for the fluid-saturated porous medium are solved analytically using the integral boundary layer approximation. This integral solution is coupled to the energy equation for the flat plate, where the longitudinal heat conduction effects are taken into account. The resulting equations are then reduced to an integro-differential equation which is solved by regular perturbation techniques and numerical methods. The analytical and numerical predictions for the temperature profile of the plate and appropriate local and average Nusselt numbers are plotted for finite values of the conduction parameter, {alpha}, which represents the presence of the longitudinal heat conduction effects. (authors)

  8. The influence of product differentiation strategy on operational performance at Small and Medium Enterprises (SMEs in South Sulawesi, Indonesia

    Directory of Open Access Journals (Sweden)

    Muhammad Yunus Amar

    2015-12-01

    Full Text Available In the last decade, many researchers have conducted studies on the efforts to improve corporate performance through the stimulation of specific business strategy approach. This study aims to analyze the effect of product differentiation strategy on operating performance of the company. The study was conducted on industrial of SMEs in South Sulawesi, Indonesia using a survey method with the sample of 75 respondents. The data were collected through questionnaires, and processed by the method of path analysis. The results show that the strategy of product differentiation (vertical and horizontal affects the operational performance of industrial of the SMEs significantly and negatively. It has implications such as in the early stages of the implementation of this strategy; the company can issue additional production costs in the form of material costs, and more failing products without being accompanied by an increase in new customers. This study can be continued to further examine the relationship of differentiation strategy implementation and performance of the company involving a moderator variable lag-time and the role of production technology in the research model.

  9. Medium Range Ensembles Flood Forecasts for Community Level Applications

    Science.gov (United States)

    Fakhruddin, S.; Kawasaki, A.; Babel, M. S.; AIT

    2013-05-01

    Early warning is a key element for disaster risk reduction. In recent decades, there has been a major advancement in medium range and seasonal forecasting. These could provide a great opportunity to improve early warning systems and advisories for early action for strategic and long term planning. This could result in increasing emphasis on proactive rather than reactive management of adverse consequences of flood events. This can be also very helpful for the agricultural sector by providing a diversity of options to farmers (e.g. changing cropping pattern, planting timing, etc.). An experimental medium range (1-10 days) flood forecasting model has been developed for Bangladesh which provides 51 set of discharge ensembles forecasts of one to ten days with significant persistence and high certainty. This could help communities (i.e. farmer) for gain/lost estimation as well as crop savings. This paper describe the application of ensembles probabilistic flood forecast at the community level for differential decision making focused on agriculture. The framework allows users to interactively specify the objectives and criteria that are germane to a particular situation, and obtain the management options that are possible, and the exogenous influences that should be taken into account before planning and decision making. risk and vulnerability assessment was conducted through community consultation. The forecast lead time requirement, users' needs, impact and management options for crops, livestock and fisheries sectors were identified through focus group discussions, informal interviews and questionnaire survey.

  10. Alteration of microRNA expression of human dental pulp cells during odontogenic differentiation.

    Science.gov (United States)

    Gong, Qimei; Wang, Runfu; Jiang, Hongwei; Lin, Zhengmei; Ling, Junqi

    2012-10-01

    MicroRNAs (miRNAs) play momentous roles in various biological processes including cell differentiation. However, little is known about the role of miRNAs in human dental pulp cells (hDPCs) during odontogenic differentiation. The aims of this study were to investigate the expression of miRNAs in the primary culture of hDPCs when incubated in odontogenic medium. The potential characteristics of hDPCs were investigated by miRNA microarray and real-time reverse transcriptase polymerase chain reaction. Bioinformatics (ie, target prediction, Gene Ontology analysis, and Kyoto Encyclopedia of Genes and Genomes mapping tools) were applied for predicting the complementary target genes of miRNAs and their biological functions. A total of 22 miRNAs were differentially expressed in which 12 miRNAs up-regulated and 10 miRNAs down-regulated in differentiated hDPCs compared with the control. The target genes of differential miRNAs were predicted to associate with several biological functions and signaling pathways including the mitogen-activated protein kinase (MAPK) and the Wnt signaling pathway. The differential expression miRNAs may be involved in governing hDPC odontogenic differentiation, thus contributing to the future investigations of regulatory mechanisms in reparative dentin formation and dental pulp regeneration. Copyright © 2012 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  11. Distinct gene expression signatures in human embryonic stem cells differentiated towards definitive endoderm at single-cell level

    DEFF Research Database (Denmark)

    Norrman, Karin; Strömbeck, Anna; Semb, Henrik

    2013-01-01

    for the three activin A based protocols applied. Our data provide novel insights in DE gene expression at the cellular level of in vitro differentiated human embryonic stem cells, and illustrate the power of using single-cell gene expression profiling to study differentiation heterogeneity and to characterize...... of anterior definitive endoderm (DE). Here, we differentiated human embryonic stem cells towards DE using three different activin A based treatments. Differentiation efficiencies were evaluated by gene expression profiling over time at cell population level. A panel of key markers was used to study DE...... formation. Final DE differentiation was also analyzed with immunocytochemistry and single-cell gene expression profiling. We found that cells treated with activin A in combination with sodium butyrate and B27 serum-free supplement medium generated the most mature DE cells. Cell population studies were...

  12. BIFIDUM-MEDIUM FOR ISOLATION AND CULTIVATION OF BIFIDOBACTERIA

    Directory of Open Access Journals (Sweden)

    L. V. Domotenko

    2014-01-01

    Full Text Available The comparative evaluation of Bifidum-medium quality (the nutrient medium for cultivation and selection bifidobacteria, dry and Blaurock medium has been performed. It was shown that Bifidum-medium supports the typical growth of the main types of bifidobacteria: Bifidobacterium bifidum, B. breve, B. adolescentis, B. infantis, B. longum. The comparable results for the efficacy and the activity accumulation of acid were obtained in the study of probiotic «bifidumbacterin » using both media. In studies of faecal material the medium concentration of bifidobacteria was found 8.15 lg CFU/g on Bifidum-medium and 6.68 lg CFU/g on Blaurock medium.

  13. Interplanetary medium and geomagnetic activity after compact flare triplets 1966-1981

    International Nuclear Information System (INIS)

    Ivanov, K.G.; Mikerina, N.V.; Pavlov, P.P.

    1986-01-01

    The interplanetary medium state and geomagnetic activity when the Earth is getting into this or that interplanetary disturbance zone after flare triplets, i.e. trains of three solar flares out of an active zone, are considered. There are the following conditionally differentiated zones in the interplanetary disturbance configuration: a forbidden (F), a perturbed (P) and a normal (N) zones of interplanetary disturbance. The interplanetary medium disturbances and geomagnetic activity after trains of three flares of class 2 and higher out of one of active zones depend on the following factors: the magnetic axis orientation of a bipolar group of active zone spots appeared after flares, time interval between the first and second flares in the train, flare intensity. The conditions of maximum disturbance occurrence pointed out. The interplanetary and geomagnetic disturbance intensity in the N zone is higher than that of the F and P zones (i.e. in the proximity of the great circle planes passing through the flares parallel with tha active zone magnetic axes), and it is higher after quasicompact rather than after compact triplets (i.e. it considerably grows when passing over the critical value of the time interval betwenn the first and second triplet flares, τ 12 =16 h)

  14. A combined disc method with resazurin agar plate assay for early phenotypic screening of KPC, MBL and OXA-48 carbapenemases among Enterobacteriaceae.

    Science.gov (United States)

    Teethaisong, Y; Eumkeb, G; Nakouti, I; Evans, K; Hobbs, G

    2016-08-01

    To validate a combined disc method along with resazurin chromogenic agar for early screening and differentiation of Klebsiella pneumoniae carbapenemase, metallo-β-lactamase and OXA-48 carbapenemase-producing Enterobacteriaceae. The combined disc test comprising of meropenem alone and with EDTA, phenylboronic acid or both EDTA and phenylboronic acid, and temocillin alone were evaluated with the resazurin chromogenic agar plate assay against a total of 86 molecularly confirmed Enterobacteriaceae clinical isolates (11 metallo-β-lactamases, eight Kl. pneumoniae carbapenemases, 11 OXA-48, 32 AmpC and 15 extended-spectrum-β-lactamase producers and nine co-producers of extended-spectrum-β-lactamase and AmpC). The inhibition zone diameters were measured and interpreted at 7 h for the presence of carbapenemase. All carbapenemase producers were phenotypically distinguished by this assay with 100% sensitivity and specificity. This early phenotypic method is very simple, inexpensive, and reliable in the detection and differentiation of carbapenemase-producing Enterobacteriaceae. It could be exploited in any microbiological laboratory for diagnosis of these recalcitrant bacteria. This assay poses excellent performance in discrimination of Kl. pneumoniae carbapenemase, metallo-β-lactamase and OXA-48 carbapenemases within 7 h, which is much faster than conventional disc diffusion methods. The rapid detection could help clinicians screen patients, control infection and provide epidemiological surveillance. © 2016 The Society for Applied Microbiology.

  15. Living biointerfaces based on non-pathogenic bacteria support stem cell differentiation

    Science.gov (United States)

    Hay, Jake J.; Rodrigo-Navarro, Aleixandre; Hassi, Karoliina; Moulisova, Vladimira; Dalby, Matthew J.; Salmeron-Sanchez, Manuel

    2016-02-01

    Lactococcus lactis, a non-pathogenic bacteria, has been genetically engineered to express the III7-10 fragment of human fibronectin as a membrane protein. The engineered L. lactis is able to develop biofilms on different surfaces (such as glass and synthetic polymers) and serves as a long-term substrate for mammalian cell culture, specifically human mesenchymal stem cells (hMSC). This system constitutes a living interface between biomaterials and stem cells. The engineered biofilms remain stable and viable for up to 28 days while the expressed fibronectin fragment induces hMSC adhesion. We have optimised conditions to allow long-term mammalian cell culture, and found that the biofilm is functionally equivalent to a fibronectin-coated surface in terms of osteoblastic differentiation using bone morphogenetic protein 2 (BMP-2) added to the medium. This living bacteria interface holds promise as a dynamic substrate for stem cell differentiation that can be further engineered to express other biochemical cues to control hMSC differentiation.

  16. Differential Optical-absorption Spectroscopy (doas) System For Urban Atmospheric-pollution Monitoring

    OpenAIRE

    Edner, H; Ragnarson, P; Spannare, S; Svanberg, Sune

    1993-01-01

    We describe a fully computer-controlled differential optical absorption spectroscopy system for atmospheric air pollution monitoring. A receiving optical telescope can sequentially tune in to light beams from a number of distant high-pressure Xe lamp light sources to cover the area of a medium-sized city. A beam-finding servosystem and automatic gain control permit unattended long-time monitoring. Using an astronomical code, we can also search and track celestial sources. Selected wavelength ...

  17. Anisotropic scattering in three dimensional differential approximation of radiation heat transfer

    International Nuclear Information System (INIS)

    Condiff, D.W.

    1987-01-01

    The differential approximation is extended to account for anisotropic scattering in invariant three dimensional form. A moment method using polyadic Legendre functions establishes that pressure cross sections should take precedence over extinction cross sections for treating radiation heat transfer in an absorbing, emitting, and scattering medium, and that use of these cross sections accounts for the extent of preferred forward or backwards scattering. The procedure and principle is extended to polyadic P-N approximations

  18. Heat transfer in porous medium embedded with vertical plate: Non-equilibrium approach - Part B

    Energy Technology Data Exchange (ETDEWEB)

    Quadir, G. A., E-mail: Irfan-magami@Rediffmail.com, E-mail: gaquadir@gmail.com [School of Mechatronic Engineering, University Malaysia Perlis, Pauh Putra, 02600 Arau, Perlis (Malaysia); Badruddin, Irfan Anjum [Dept. of Mechanical Engineering, University of Malaya, Kuala Lumpur, 50603 (Malaysia)

    2016-06-08

    This work is continuation of the paper Part A. Due to large number of results, the paper is divided into two section with section-A (Part A) discussing the effect of various parameters such as heat transfer coefficient parameter, thermal conductivity ratio etc. on streamlines and isothermal lines. Section-B highlights the heat transfer characteristics in terms of Nusselt number The Darcy model is employed to simulate the flow inside the medium. It is assumed that the heat transfer takes place by convection and radiation. The governing partial differential equations are converted into non-dimensional form and solved numerically using finite element method.

  19. Approximation and stability of three-dimensional natural convection flows in a porous medium

    International Nuclear Information System (INIS)

    Janotto, Marie-Laurence

    1991-01-01

    The equations of the three-dimensional natural convection in a porous medium within a differentially heated horizontal walls cavity are solved by a pseudo-spectral method. First we will present the evolution of the two main modes according to two models of convection. A few asymptotic properties connected to the small and large eddies are set up and numerically validated. A new approximate inertial manifold is then proposed. The numerical scheme used is an exponential fitting algorithm the convergence of which is proved. We will present the physical mechanism at the origin of the un-stationary three-dimensional convection at high Rayleigh numbers. (author) [fr

  20. [Distribution of Candida species in vaginal specimens and evaluation of CHROMagar Candida medium].

    Science.gov (United States)

    Gültekin, Berna; Yazici, Vesile; Aydin, Neriman

    2005-07-01

    Identification of Candida species is important to guide treatment in vulvovaginal candidiasis which is seen frequently and needs long-term therapy due to recurrence. The aim of this study was to determine the species distribution of Candida isolated from vaginal specimens and evaluation of CHROMagar Candida medium in the laboratory diagnosis. Samples from 80 patients who were clinically diagnosed as vaginitis have been analysed in our laboratory. Colonies appeared on CHROMagar Candida media after 48 hours of incubation at 35 degrees C were evaluated for their colors and characteristics. Candida strains were identified by germ tube test, growth on corn meal Tween 80 agar and when necessary also by API 20 C AUX commercial kit. A total of 84 Candida strains were isolated from 80 patients. Two different Candida species have been isolated from four (5%) of the samples. Among Candida strains isolated, 45 (53.6%) were C. albicans, 29 (34.5%) C. glabrata, 7 (8.3%) C. krusei, and 3 (3.6%) C. kefyr. All of the C. albicans and six of the seven C. krusei isolates have been identified correctly by CHROMagar Candida medium. These results showed that C. albicans is still the most frequently isolated species from vaginal samples. It was concluded that CHROMagar Candida medium is useful for identification of colonies due to frequently seen Candida species and also in differentiation of multiple Candida species grown on the same culture.

  1. Quantitative real-time RT-PCR and chromogenic in situ hybridization: precise methods to detect HER-2 status in breast carcinoma

    International Nuclear Information System (INIS)

    Rosa, Fabíola E; Silveira, Sara M; Silveira, Cássia GT; Bérgamo, Nádia A; Neto, Francisco A Moraes; Domingues, Maria AC; Soares, Fernando A; Caldeira, José RF; Rogatto, Silvia R

    2009-01-01

    HER-2 gene testing has become an integral part of breast cancer patient diagnosis. The most commonly used assay in the clinical setting for evaluating HER-2 status is immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH). These procedures permit correlation between HER-2 expression and morphological features. However, FISH signals are labile and fade over time, making post-revision of the tumor difficult. CISH (chromogenic in situ hybridization) is an alternative procedure, with certain advantages, although still limited as a diagnostic tool in breast carcinomas. To elucidate the molecular profile of HER-2 status, mRNA and protein expression in 75 invasive breast carcinomas were analyzed by real time quantitative RT-PCR (qRT-PCR) and IHC, respectively. Amplifications were evaluated in 43 of these cases by CISH and in 11 by FISH. The concordance rate between IHC and qRT-PCR results was 78.9%, and 94.6% for qRT-PCR and CISH. Intratumoral heterogeneity of HER-2 status was identified in three cases by CISH. The results of the three procedures were compared and showed a concordance rate of 83.8%; higher discordances were observed in 0 or 1+ immunostaining cases, which showed high-level amplification (15.4%) and HER-2 transcript overexpression (20%). Moreover, 2+ immunostaining cases presented nonamplified status (50%) by CISH and HER-2 downexpression (38.5%) by qRT-PCR. In general, concordance occurred between qRT-PCR and CISH results. A high concordance was observed between CISH/qRT-PCR and FISH. Comparisons with clinicopathological data revealed a significant association between HER-2 downexpression and the involvement of less than four lymph nodes (P = 0.0350). Based on these findings, qRT-PCR was more precise and reproducible than IHC. Furthermore, CISH was revealed as an alternative and useful procedure for investigating amplifications involving the HER-2 gene

  2. Differential conservation and divergence of fertility genes boule and dazl in the rainbow trout.

    Directory of Open Access Journals (Sweden)

    Mingyou Li

    Full Text Available BACKGROUND: The genes boule and dazl are members of the DAZ (Deleted in Azoospermia family encoding RNA binding proteins essential for germ cell development. Although dazl exhibits bisexual expression in mitotic and meiotic germ cells in diverse animals, boule shows unisexual meiotic expression in invertebrates and mammals but a bisexual mitotic and meiotic expression in medaka. How boule and dazl have evolved different expression patterns in diverse organisms has remained unknown. METHODOLOGY AND PRINCIPAL FINDINGS: Here we chose the fish rainbow trout (Oncorhynchus mykiss as a second lower vertebrate model to investigate the expression of boule and dazl. By molecular cloning and sequence comparison, we identified cDNAs encoding the trout Boule and Dazl proteins, which have a conserved RNA-recognition motif and a maximal similarity to their homologs. By RT-PCR analysis, adult RNA expression of trout boule and dazl is restricted to the gonads of both sexes. By chromogenic and two-color fluorescence in situ hybridization, we revealed bisexual and germline-specific expression of boule and dazl. We found that dazl displays conserved expression throughout gametogenesis and concentrates in the Balbinani's body of early oocytes and the chromatoid body of sperm. Surprisingly, boule exhibits mitotic and meiotic expression in the male but meiosis-specific expression in the female. CONCLUSIONS: Our data underscores differential conservation and divergence of DAZ family genes during vertebrate evolution. We propose a model in which the diversity of boule expression in sex and stage specificity might have resulted from selective loss or gain of its expression in one sex and mitotic germ cells.

  3. BAG3 is involved in neuronal differentiation and migration.

    Science.gov (United States)

    Santoro, Antonietta; Nicolin, Vanessa; Florenzano, Fulvio; Rosati, Alessandra; Capunzo, Mario; Nori, Stefania L

    2017-05-01

    Bcl2-associated athanogene 3 (BAG3) protein belongs to the family of co-chaperones interacting with several heat shock proteins. It plays a key role in protein quality control and mediates the clearance of misfolded proteins. Little is known about the expression and cellular localization of BAG3 during nervous system development and differentiation. Therefore, we analyze the subcellular distribution and expression of BAG3 in nerve-growth-factor-induced neurite outgrowth in PC12 cells and in developing and adult cortex of mouse brain. In differentiated PC12 cells, BAG3 was localized mainly in the neuritic domain rather than the cell body, whereas in control cells, it appeared to be confined to the cytoplasm near the nuclear membrane. Interestingly, the change of BAG3 localization during neuronal differentiation was associated only with a slight increase in total BAG3 expression. These data were coroborated by transmission electron microscopy showing that BAG3 was confined mainly within large dense-core vesicles of the axon in differentiated PC12 cells. In mouse developing cortex, BAG3 appeared to be intensely expressed in cellular processes of migrating cells, whereas in adult brain, a diffuse expression of low to medium intensity was detected in neuronal cell bodies. These findings suggest that BAG3 expression is required for neuronal differentiation and migration and that its role is linked to a change in its distribution pattern rather than to an increase in its protein expression levels.

  4. Angiofibroma of soft tissue with fibrohistiocytic features and intratumor genetic heterogeneity of NCOA2 gene rearrangement revealed by chromogenic in situ hybridization: a case report.

    Science.gov (United States)

    Fukuda, Yumiko; Motoi, Toru; Kato, Ikuma; Ikegami, Masachika; Funata, Nobuaki; Ohtomo, Rie; Horiguchi, Shinichiro; Goto, Takahiro; Hishima, Tsunekazu

    2014-05-01

    Angiofibroma of soft tissue is a recently described soft tissue tumor that is characterized by fibroblastic spindle tumor cells with arborizing capillary proliferation. Cytogenetically, it harbors a specific fusion gene involving the nuclear receptor coactivator 2 (NCOA2) gene. We report here additional new pathological and cytogenetic features. A soft tissue tumor in the left thigh of 73-year-old female was investigated. Microscopically, histiocytoid tumor cells were scattered in an edematous background with branching capillary proliferation. Immunohistochemically, we identified that the tumor cells were positive for histiocytic markers such as CD68 and CD163. Rearrangement of the NCOA2 gene was detected successfully by chromogenic in situ hybridization; however, abnormal signal patterns were observed in only a small subset of tumor cells. Unlike typical tumors with bland spindle cells, the present tumor needs to be distinguished from myxoid, dendritic and clear cell tumors. This case may suggest that angiofibroma of soft tissue is not in the center of the fibroblastic/myofibroblastic tumor group, but rather shows a fibrohistiocytic nature. We also found intratumor genetic heterogeneity, which is uncommon for a translocation-associated tumor. Therefore, careful evaluation is required to detect the gene rearrangement in this tumor entity. © 2014 The Authors. Pathology International © 2014 Japanese Society of Pathology and Wiley Publishing Asia Pty Ltd.

  5. Automatic differentiation for design sensitivity analysis of structural systems using multiple processors

    Science.gov (United States)

    Nguyen, Duc T.; Storaasli, Olaf O.; Qin, Jiangning; Qamar, Ramzi

    1994-01-01

    An automatic differentiation tool (ADIFOR) is incorporated into a finite element based structural analysis program for shape and non-shape design sensitivity analysis of structural systems. The entire analysis and sensitivity procedures are parallelized and vectorized for high performance computation. Small scale examples to verify the accuracy of the proposed program and a medium scale example to demonstrate the parallel vector performance on multiple CRAY C90 processors are included.

  6. Photonic-resonant left-handed medium

    International Nuclear Information System (INIS)

    Shen Jianqi

    2006-01-01

    A new scheme to realize simultaneously negative permittivity and permeability in a coherent atomic vapor medium (photonic-resonant material) via a coherent driving mechanism is suggested. It is verified that the atomic system coherently driven by a strong optical field will give rise to a negative refractive index in certain probe frequency ranges. One of the most remarkable features of the present scheme is such that a slab fabricated by the left-handed vapor medium is an ideal candidate for designing perfect lenses since the photonic-resonant atomic vapor cannot only exhibit an isotropic negative refractive index, but also provide a good impedance match at the air-medium interfaces

  7. Organisational Learning and the Organisational Life Cycle: The Differential Aspects of an Integrated Relationship in SMEs

    Science.gov (United States)

    Tam, Steven; Gray, David E.

    2016-01-01

    Purpose: The purpose of this study is to relate the practice of organisational learning in small- and medium-sized enterprises (SMEs) to the organisational life cycle (OLC), contextualising the differential aspects of an integrated relationship between them. Design/methodology/approach: It is a mixed-method study with two consecutive phases. In…

  8. Degradation testing of Mg alloys in Dulbecco's modified eagle medium: Influence of medium sterilization.

    Science.gov (United States)

    Marco, Iñigo; Feyerabend, Frank; Willumeit-Römer, Regine; Van der Biest, Omer

    2016-05-01

    This work studies the in vitro degradation of Mg alloys for bioabsorbable implant applications under near physiological conditions. For this purpose, the degradation behaviour of Mg alloys in Dulbecco's modified eagle medium (DMEM) which is a commonly used cell culture medium is analysed. Unfortunately, DMEM can be contaminated by microorganisms, acidifying the medium and accelerating the Mg degradation process by dissolution of protective degradation layers, such as (Mgx,Cay)(PO4)z. In this paper the influence of sterilization by applying UV-C radiation and antibiotics (penicillin/streptomycin) is analysed with two implant material candidates: Mg-Gd and Mg-Ag alloys; and pure magnesium as well as Mg-4Y-3RE as a reference. Copyright © 2016 Elsevier B.V. All rights reserved.

  9. Choice of Scottish Gaelic-Medium and Welsh-Medium Education at the Primary and Secondary School Stages: Parent and Pupil Perspectives

    Science.gov (United States)

    O'Hanlon, Fiona

    2015-01-01

    Results are presented of a comparative study of the reasons for parental choice of Scottish Gaelic-medium and Welsh-medium primary education in the year 2000 and of the reasons for pupils' decisions to continue with Gaelic or Welsh-medium education at secondary school in 2007. Parents in both contexts cited the quality of Celtic-medium education…

  10. Apc bridges Wnt/{beta}-catenin and BMP signaling during osteoblast differentiation of KS483 cells

    Energy Technology Data Exchange (ETDEWEB)

    Miclea, Razvan L., E-mail: R.L.Miclea@lumc.nl [Department of Pediatrics, Leiden University Medical Centre (LUMC), Leiden (Netherlands); Horst, Geertje van der, E-mail: G.van_der_Horst@lumc.nl [Department of Urology, LUMC, Leiden (Netherlands); Robanus-Maandag, Els C., E-mail: E.C.Robanus@lumc.nl [Department of Human Genetics, LUMC, Leiden (Netherlands); Loewik, Clemens W.G.M., E-mail: C.W.G.M.Lowik@lumc.nl [Department of Endocrinology and Metabolic Diseases, LUMC, Leiden (Netherlands); Oostdijk, Wilma, E-mail: W.Oostdijk@lumc.nl [Department of Pediatrics, Leiden University Medical Centre (LUMC), Leiden (Netherlands); Wit, Jan M., E-mail: J.M.Wit@lumc.nl [Department of Pediatrics, Leiden University Medical Centre (LUMC), Leiden (Netherlands); Karperien, Marcel, E-mail: H.B.J.Karperien@tnw.utwente.nl [MIRA Institute for Biomedical Technology and Technical Medicine, Department of Tissue Regeneration, University of Twente, Zuidhorst Room ZH 144, Drienerlolaan 5, 7522 NB Enschede (Netherlands)

    2011-06-10

    The canonical Wnt signaling pathway influences the differentiation of mesenchymal cell lineages in a quantitative and qualitative fashion depending on the dose of {beta}-catenin signaling. Adenomatous polyposis coli (Apc) is the critical intracellular regulator of {beta}-catenin turnover. To better understand the molecular mechanisms underlying the role of Apc in regulating the differentiation capacity of skeletal progenitor cells, we have knocked down Apc in the murine mesenchymal stem cell-like KS483 cells by stable expression of Apc-specific small interfering RNA. In routine culture, KSFrt-Apc{sub si} cells displayed a mesenchymal-like spindle shape morphology, exhibited markedly decreased proliferation and increased apoptosis. Apc knockdown resulted in upregulation of the Wnt/{beta}-catenin and the BMP/Smad signaling pathways, but osteogenic differentiation was completely inhibited. This effect could be rescued by adding high concentrations of BMP-7 to the differentiation medium. Furthermore, KSFrt-Apc{sub si} cells showed no potential to differentiate into chondrocytes or adipocytes. These results demonstrate that Apc is essential for the proliferation, survival and differentiation of KS483 cells. Apc knockdown blocks the osteogenic differentiation of skeletal progenitor cells, a process that can be overruled by high BMP signaling.

  11. Delta-like 1/Fetal Antigen-1 (Dlk1/FA1) Is a Novel Regulator of Chondrogenic Cell Differentiation via Inhibition of the Akt Kinase-dependent Pathway*

    Science.gov (United States)

    Chen, Li; Qanie, Diyako; Jafari, Abbas; Taipaleenmaki, Hanna; Jensen, Charlotte H.; Säämänen, Anna-Marja; Sanz, Maria Luisa Nueda; Laborda, Jorge; Abdallah, Basem M.; Kassem, Moustapha

    2011-01-01

    Delta-like 1 (Dlk1, also known as fetal antigen-1, FA1) is a member of Notch/Delta family that inhibits adipocyte and osteoblast differentiation; however, its role in chondrogenesis is still not clear. Thus, we overexpressed Dlk1/FA1 in mouse embryonic ATDC5 cells and tested its effects on chondrogenic differentiation. Dlk1/FA1 inhibited insulin-induced chondrogenic differentiation as evidenced by reduction of cartilage nodule formation and gene expression of aggrecan, collagen Type II and X. Similar effects were obtained either by using Dlk1/FA1-conditioned medium or by addition of a purified, secreted, form of Dlk1 (FA1) directly to the induction medium. The inhibitory effects of Dlk1/FA1 were dose-dependent and occurred irrespective of the chondrogenic differentiation stage: proliferation, differentiation, maturation, or hypertrophic conversion. Overexpression or addition of the Dlk1/FA1 protein to the medium strongly inhibited the activation of Akt, but not the ERK1/2, or p38 MAPK pathways, and the inhibition of Akt by Dlk1/FA1 was mediated through PI3K activation. Interestingly, inhibition of fibronectin expression by siRNA rescued the Dlk1/FA1-mediated inhibition of Akt, suggesting interaction of Dlk1/FA1 and fibronectin in chondrogenic cells. Our results identify Dlk1/FA1 as a novel regulator of chondrogenesis and suggest Dlk1/FA1 acts as an inhibitor of the PI3K/Akt pathways that leads to its inhibitory effects on chondrogenesis. PMID:21724852

  12. Catabolic factors and osteoarthritis-conditioned medium inhibit chondrogenesis of human mesenchymal stem cells.

    Science.gov (United States)

    Heldens, Genoveva T H; Blaney Davidson, Esmeralda N; Vitters, Elly L; Schreurs, B Willem; Piek, Ester; van den Berg, Wim B; van der Kraan, Peter M

    2012-01-01

    Articular cartilage has a very limited intrinsic repair capacity leading to progressive joint damage. Therapies involving tissue engineering depend on chondrogenic differentiation of progenitor cells. This chondrogenic differentiation will have to survive in a diseased joint. We postulate that catabolic factors in this environment inhibit chondrogenesis of progenitor cells. We investigated the effect of a catabolic environment on chondrogenesis in pellet cultures of human mesenchymal stem cells (hMSCs). We exposed chondrogenically differentiated hMSC pellets, to interleukin (IL)-1α, tumor necrosis factor (TNF)-α or conditioned medium derived from osteoarthritic synovium (CM-OAS). IL-1α and TNF-α in CM-OAS were blocked with IL-1Ra or Enbrel, respectively. Chondrogenesis was determined by chondrogenic markers collagen type II, aggrecan, and the hypertrophy marker collagen type X on mRNA. Proteoglycan deposition was analyzed by safranin o staining on histology. IL-1α and TNF-α dose-dependently inhibited chondrogenesis when added at onset or during progression of differentiation, IL-1α being more potent than TNF-α. CM-OAS inhibited chondrogenesis on mRNA and protein level but varied in extent between patients. Inhibition of IL-1α partially overcame the inhibitory effect of the CM-OAS on chondrogenesis whereas the TNF-α contribution was negligible. We show that hMSC chondrogenesis is blocked by either IL-1α or TNF-α alone, but that there are additional factors present in CM-OAS that contribute to inhibition of chondrogenesis, demonstrating that catabolic factors present in OA joints inhibit chondrogenesis, thereby impairing successful tissue engineering.

  13. Medium Modifications of Hadrons in Photon Induced Reactions

    International Nuclear Information System (INIS)

    Schadmand, S.

    2004-01-01

    Indications for in-medium modifications of hadron properties are reported from photoabsorption and meson production experiments. Strong medium modifications are observed in inclusive photoabsorption experiments and theoretical models investigate the in-medium dynamics of baryon resonances and their coupling to mesons. Recent experiments study the in-medium behavior of scalar and vector mesons where theoretical models expect in-medium modifications of the meson spectral functions that might be connected to partial restoration of chiral symmetry

  14. Fiscal 1980 Sunshine Project research report. Development of hydrothermal power plant. Development of binary cycle power plant (Research on heat cycle, heat medium, material and heat medium turbine); 1980 nendo nessui riyo hatsuden plant no kaihatsu seika hokokusho. Binary cycle hatsuden plant no kaihatsu (netsu cycle oyobi netsubaitai no kenkyu, zairyo no kenkyu narabini netsubaitai turbine no kenkyu)

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1981-03-01

    This report summarizes the fiscal 1980 research result on each element of the next 10MW class geothermal binary cycle power plant, following last year. In the research on heat cycle and heat medium, measurement was made on the liquid density, vapor density, liquid specific heat, vapor specific heat and thermal conductivity of 8 heat media to prepare the precise pressure enthalpy chart. The thermal stability of each medium was also measured under a flow condition. The heat cycle of each medium was calculated in a hydrothermal temperature range of 120-160 degrees C for evaluation of its output. In the research on material, field corrosion test and laboratory simulation were made on 3 kinds of heat exchanger martials for acidic hot water to study the corrosion behavior of welding members. In the research on heat medium turbine, study was made on sealing characteristics such as differential pressure, flow rate and friction of sealing oil for oil film seal and mechanical seal as shaft seal devices of heat medium turbines for the 10MW class geothermal plant. (NEDO)

  15. Steady Boundary Layer Slip Flow along with Heat and Mass Transfer over a Flat Porous Plate Embedded in a Porous Medium

    Science.gov (United States)

    Aziz, Asim; Siddique, J. I.; Aziz, Taha

    2014-01-01

    In this paper, a simplified model of an incompressible fluid flow along with heat and mass transfer past a porous flat plate embedded in a Darcy type porous medium is investigated. The velocity, thermal and mass slip conditions are utilized that has not been discussed in the literature before. The similarity transformations are used to transform the governing partial differential equations (PDEs) into a nonlinear ordinary differential equations (ODEs). The resulting system of ODEs is then reduced to a system of first order differential equations which was solved numerically by using Matlab bvp4c code. The effects of permeability, suction/injection parameter, velocity parameter and slip parameter on the structure of velocity, temperature and mass transfer rates are examined with the aid of several graphs. Moreover, observations based on Schmidt number and Soret number are also presented. The result shows, the increase in permeability of the porous medium increase the velocity and decrease the temperature profile. This happens due to a decrease in drag of the fluid flow. In the case of heat transfer, the increase in permeability and slip parameter causes an increase in heat transfer. However for the case of increase in thermal slip parameter there is a decrease in heat transfer. An increase in the mass slip parameter causes a decrease in the concentration field. The suction and injection parameter has similar effect on concentration profile as for the case of velocity profile. PMID:25531301

  16. Steady boundary layer slip flow along with heat and mass transfer over a flat porous plate embedded in a porous medium.

    Science.gov (United States)

    Aziz, Asim; Siddique, J I; Aziz, Taha

    2014-01-01

    In this paper, a simplified model of an incompressible fluid flow along with heat and mass transfer past a porous flat plate embedded in a Darcy type porous medium is investigated. The velocity, thermal and mass slip conditions are utilized that has not been discussed in the literature before. The similarity transformations are used to transform the governing partial differential equations (PDEs) into a nonlinear ordinary differential equations (ODEs). The resulting system of ODEs is then reduced to a system of first order differential equations which was solved numerically by using Matlab bvp4c code. The effects of permeability, suction/injection parameter, velocity parameter and slip parameter on the structure of velocity, temperature and mass transfer rates are examined with the aid of several graphs. Moreover, observations based on Schmidt number and Soret number are also presented. The result shows, the increase in permeability of the porous medium increase the velocity and decrease the temperature profile. This happens due to a decrease in drag of the fluid flow. In the case of heat transfer, the increase in permeability and slip parameter causes an increase in heat transfer. However for the case of increase in thermal slip parameter there is a decrease in heat transfer. An increase in the mass slip parameter causes a decrease in the concentration field. The suction and injection parameter has similar effect on concentration profile as for the case of velocity profile.

  17. Reliability of chromogenic in situ hybridization for epidermal growth factor receptor gene copy number detection in non-small-cell lung carcinomas: a comparison with fluorescence in situ hybridization study.

    Science.gov (United States)

    Yoo, Seol Bong; Lee, Hyun Ju; Park, Jung Ok; Choe, Gheeyoung; Chung, Doo Hyun; Seo, Jeong-Wook; Chung, Jin-Haeng

    2010-03-01

    Fluorescence in situ hybridization (FISH) has been known to be the most representative and standardized test for assessing gene amplification. However, FISH requires a fluorescence microscope, the signals are labile and rapidly fade over time. Recently, chromogenic in situ hybridization (CISH) has emerged as a potential alternative to FISH. The aim of this study is to test the reliability of CISH technique for the detection of epidermal growth factor receptor (EGFR) gene amplification in non-small-cell lung carcinomas (NSCLC), to compare CISH results with FISH. A total of 277 formalin-fixed and paraffin embedded NSCLC tissue samples were retrieved from the surgical pathology archives at Seoul National University Bundang Hospital. CISH and FISH examinations were performed to test EGFR gene amplification status. There was high concordance in the assessment of EGFR gene copy number between CISH and FISH tests (Kappa coefficient=0.83). Excellent concordance was shown between two observers on the interpretation of the CISH results (Kappa coefficient=0.90). In conclusion, CISH result is highly reproducible, accurate and practical method to determine EGFR gene amplification in NSCLC. In addition, CISH allows a concurrent analysis of histological features of the tumors and gene copy numbers.

  18. Three-dimensional neural differentiation of embryonic stem cells with ACM induction in microfibrous matrices in bioreactors.

    Science.gov (United States)

    Liu, Ning; Ouyang, Anli; Li, Yan; Yang, Shang-Tian

    2013-01-01

    The clinical use of pluripotent stem cell (PSC)-derived neural cells requires an efficient differentiation process for mass production in a bioreactor. Toward this goal, neural differentiation of murine embryonic stem cells (ESCs) in three-dimensional (3D) polyethylene terephthalate microfibrous matrices was investigated in this study. To streamline the process and provide a platform for process integration, the neural differentiation of ESCs was induced with astrocyte-conditioned medium without the formation of embryoid bodies, starting from undifferentiated ESC aggregates expanded in a suspension bioreactor. The 3D neural differentiation was able to generate a complex neural network in the matrices. When compared to 2D differentiation, 3D differentiation in microfibrous matrices resulted in a higher percentage of nestin-positive cells (68% vs. 54%) and upregulated gene expressions of nestin, Nurr1, and tyrosine hydroxylase. High purity of neural differentiation in 3D microfibrous matrix was also demonstrated in a spinner bioreactor with 74% nestin + cells. This study demonstrated the feasibility of a scalable process based on 3D differentiation in microfibrous matrices for the production of ESC-derived neural cells. © 2013 American Institute of Chemical Engineers.

  19. Mapping of moveout in a TTI medium

    KAUST Repository

    Stovas, A.; Alkhalifah, Tariq Ali

    2012-01-01

    To compute moveout in a transversely isotropic medium with tilted symmetry axis is a very complicated problem. We propose to split this problem into two parts. First, to compute the moveout in a corresponding VTI medium. Second, to map the computed moveout to a TTI medium.

  20. Fractional diffusion equation for heterogeneous medium

    International Nuclear Information System (INIS)

    Polo L, M. A.; Espinosa M, E. G.; Espinosa P, G.; Del Valle G, E.

    2011-11-01

    The asymptotic diffusion approximation for the Boltzmann (transport) equation was developed in 1950 decade in order to describe the diffusion of a particle in an isotropic medium, considers that the particles have a diffusion infinite velocity. In this work is developed a new approximation where is considered that the particles have a finite velocity, with this model is possible to describe the behavior in an anomalous medium. According with these ideas the model was obtained from the Fick law, where is considered that the temporal term of the current vector is not negligible. As a result the diffusion equation of fractional order which describes the dispersion of particles in a highly heterogeneous or disturbed medium is obtained, i.e., in a general medium. (Author)

  1. Optical illusions induced by rotating medium

    Science.gov (United States)

    Zang, XiaoFei; Huang, PengCheng; Zhu, YiMing

    2018-03-01

    Different from the traditional single-function electromagnetic wave rotators (rotate the electromagnetic wavefronts), we propose that rotating medium can be extended to optical illusions such as breaking the diffraction limit and overlapping illusion. Furthermore, the homogeneous but anisotropic rotating medium is simplified by homogeneous and isotropic positive-index materials according to the effective medium theory, which is helpful for future device fabrication. Finite element simulations for the two-dimensional case are performed to demonstrate these properties.

  2. Mesons in the nuclear Medium

    CERN Document Server

    Kotulla, M

    2006-01-01

    We discuss recent experimental results on the modification of hadron properties in a nuclear medium. Particular emphasis is placed on an $\\omega$ production experiment performed by the CBELSA/TAPS collaboration at the ELSA accelerator. The data shows a smaller $\\omega$ meson mass together with a significant increase of its width in the nuclear medium.

  3. Human platelet lysate successfully promotes proliferation and subsequent chondrogenic differentiation of adipose-derived stem cells: a comparison with articular chondrocytes.

    Science.gov (United States)

    Hildner, F; Eder, M J; Hofer, K; Aberl, J; Redl, H; van Griensven, M; Gabriel, C; Peterbauer-Scherb, A

    2015-07-01

    Fetal calf serum (FCS) bears a potential risk for carrying diseases and eliciting immune reactions. Nevertheless, it still represents the gold standard as medium supplement in cell culture. In the present study, human platelet lysate (PL) was tested as an alternative to FCS for the expansion and subsequent chondrogenic differentiation of human adipose-derived stem cells (ASCs). ASCs were expanded with 10% FCS (group F) or 5% PL (group P). Subsequently, three-dimensional (3D) micromass pellets were created and cultured for 5 weeks in chondrogenic differentiation medium. Additionally, the de- and redifferentiation potential of human articular chondrocytes (HACs) was evaluated and compared to ASCs. Both HACs and ASCs cultured with PL showed strongly enhanced proliferation rates. Redifferentiation of HACs was possible for cells expanded up to 3.3 population doublings (PD). At this stage, PL-expanded HACs demonstrated better redifferentiation potential than FCS-expanded cells. ASCs could also be differentiated following extended passaging. Glycosaminoglycan (GAG) quantification and qRT-PCR of 10 cartilage related markers demonstrated a tendency for increased chondrogenic differentiation of PL-expanded ASCs compared to cells expanded with FCS. Histologically, collagen type II but also collagen type X was mainly present in group P. The present study demonstrates that PL strongly induces proliferation of ASCs, while the chondrogenic differentiation potential is retained. HACs also showed enhanced proliferation and even better redifferentiation when previously expanded with PL. This suggests that PL is superior to FCS as a supplement for the expansion of ASCs and HACs, particularly with regard to chondrogenic (re)differentiation. Copyright © 2013 John Wiley & Sons, Ltd.

  4. Incorporation of medium chain fatty acids into fish oil triglycerides by chemical and enzymatic inter esterification

    Energy Technology Data Exchange (ETDEWEB)

    Feltes, M. M. C.; Oliveira de Pilot, L.; Gomes Correira, F.; Grimaldi, R.; Mara Block, J.; Ninow, J. L.

    2009-07-01

    Structured triglycerides (STs) containing both medium chain fatty acids (MCFA) and polyunsaturated fatty acids (PUFA) in the same molecule offer nutritional and therapeutic benefits. The aim of this work was to establish the incorporation of MCFA into fish oil triglycerides (TAGs), while maintaining substantial levels of docosahexaenoic and eicosapentaenoic acids. The effects of different acyl donors (capric acid methyl ester/MeC10 or medium chain triglyceride/TCM) and of the catalyst (chemical or enzymatic) on the fatty acid composition of the reaction product were studied. The fatty acid composition of the fish oil TAG was modified after inter esterification to contain MCFA, and it depended on the catalyst and on the substrates. Thermo grams obtained by Differential Scanning Calorimetry (DSC) showed that inter esterification promoted noteworthy changes in the melting profile of the samples. STs of clinical nutrition interest containing both EPA and DHA obtained from fish oil along with MCFA were successfully produced. (Author) 70 refs.

  5. Three dimensional vibration and bending analysis of carbon nanotubes embedded in elastic medium based on theory of elasticity

    Directory of Open Access Journals (Sweden)

    M. Shaban

    Full Text Available This paper studies free vibration and bending behavior of singlewalled carbon nanotubes (SWCNTs embedded on elastic medium based on three-dimensional theory of elasticity. To accounting the size effect of carbon nanotubes, non-local theory is adopted to shell model. The nonlocal parameter is incorporated into all constitutive equations in three dimensions. The surrounding medium is modeled as two-parameter elastic foundation. By using Fourier series expansion in axial and circumferential direction, the set of coupled governing equations are reduced to the ordinary differential equations in thickness direction. Then, the state-space method as an efficient and accurate method is used to solve the resulting equations analytically. Comprehensive parametric studies are carried out to show the influences of the nonlocal parameter, radial and shear elastic stiffness, thickness-to-radius ratio and radiusto-length ratio.

  6. Selection of culture medium and conditions for the production of ...

    African Journals Online (AJOL)

    defined medium–A, defined medium-B, synthetic medium, rich medium and industrial medium) showed that the synthetic medium yielded maximum yeast biomass (12.8 g/LDCW) followed by rich medium (11.7 g/L DCW) and defined medium B ...

  7. Medium energy nucleon-nucleus scattering theory by semi-classical distorted wave approximation

    Energy Technology Data Exchange (ETDEWEB)

    Ogata, Kazuyuki [Kyushu Univ., Fukuoka (Japan)

    1998-07-01

    The semiclassical distorted wave model (SCDW) is one of the quantum mechanical models for nucleon inelastic and charge exchange scattering at intermediate energies. SCDW can reproduce the double differential inclusive cross sections for multi-step direct processes quite well in the angular and outgoing energy regions where the model is expected to work. But the model hitherto assumed on-the-energy-shell (on-shell) nucleon-nucleon scattering in the nucleus, neglecting the difference in the distorting potentials for the incoming and the outgoing particles and also the Q-value in the case of (p,n) reactions. There had also been a problem in the treatment of the exchange of colliding nucleons. Now we modify the model to overcome those problems and put SCDW on sounder theoretical foundations. The modification results in slight reduction (increase) of double differential cross sections at forward (backward) angles. We also examine the effect of the in-medium modification of N-N cross sections in SCDW and find it small. A remedy of the disagreement at very small and large angles in terms of the Wigner transform of the single particle density matrix is also discussed. This improvement gives very promising results. (author)

  8. Brain stem slice conditioned medium contains endogenous BDNF and GDNF that affect neural crest boundary cap cells in co-culture.

    Science.gov (United States)

    Kaiser, Andreas; Kale, Ajay; Novozhilova, Ekaterina; Siratirakun, Piyaporn; Aquino, Jorge B; Thonabulsombat, Charoensri; Ernfors, Patrik; Olivius, Petri

    2014-05-30

    Conditioned medium (CM), made by collecting medium after a few days in cell culture and then re-using it to further stimulate other cells, is a known experimental concept since the 1950s. Our group has explored this technique to stimulate the performance of cells in culture in general, and to evaluate stem- and progenitor cell aptitude for auditory nerve repair enhancement in particular. As compared to other mediums, all primary endpoints in our published experimental settings have weighed in favor of conditioned culture medium, where we have shown that conditioned culture medium has a stimulatory effect on cell survival. In order to explore the reasons for this improved survival we set out to analyze the conditioned culture medium. We utilized ELISA kits to investigate whether brain stem (BS) slice CM contains any significant amounts of brain-derived neurotrophic factor (BDNF) and glial cell derived neurotrophic factor (GDNF). We further looked for a donor cell with progenitor characteristics that would be receptive to BDNF and GDNF. We chose the well-documented boundary cap (BC) progenitor cells to be tested in our in vitro co-culture setting together with cochlear nucleus (CN) of the BS. The results show that BS CM contains BDNF and GDNF and that survival of BC cells, as well as BC cell differentiation into neurons, were enhanced when BS CM were used. Altogether, we conclude that BC cells transplanted into a BDNF and GDNF rich environment could be suitable for treatment of a traumatized or degenerated auditory nerve. Copyright © 2014 Elsevier B.V. All rights reserved.

  9. High Glucose Inhibits Neural Stem Cell Differentiation Through Oxidative Stress and Endoplasmic Reticulum Stress.

    Science.gov (United States)

    Chen, Xi; Shen, Wei-Bin; Yang, Penghua; Dong, Daoyin; Sun, Winny; Yang, Peixin

    2018-06-01

    Maternal diabetes induces neural tube defects by suppressing neurogenesis in the developing neuroepithelium. Our recent study further revealed that high glucose inhibited embryonic stem cell differentiation into neural lineage cells. However, the mechanism whereby high glucose suppresses neural differentiation is unclear. To investigate whether high glucose-induced oxidative stress and endoplasmic reticulum (ER) stress lead to the inhibition of neural differentiation, the effect of high glucose on neural stem cell (the C17.2 cell line) differentiation was examined. Neural stem cells were cultured in normal glucose (5 mM) or high glucose (25 mM) differentiation medium for 3, 5, and 7 days. High glucose suppressed neural stem cell differentiation by significantly decreasing the expression of the neuron marker Tuj1 and the glial cell marker GFAP and the numbers of Tuj1 + and GFAP + cells. The antioxidant enzyme superoxide dismutase mimetic Tempol reversed high glucose-decreased Tuj1 and GFAP expression and restored the numbers of neurons and glial cells differentiated from neural stem cells. Hydrogen peroxide treatment imitated the inhibitory effect of high glucose on neural stem cell differentiation. Both high glucose and hydrogen peroxide triggered ER stress, whereas Tempol blocked high glucose-induced ER stress. The ER stress inhibitor, 4-phenylbutyrate, abolished the inhibition of high glucose or hydrogen peroxide on neural stem cell differentiation. Thus, oxidative stress and its resultant ER stress mediate the inhibitory effect of high glucose on neural stem cell differentiation.

  10. Amniocar as a proliferative medium for mesenchymal cells

    Directory of Open Access Journals (Sweden)

    V. V. Chestkov

    2014-01-01

    Full Text Available Objectives. To develop the Amniocar nutrient medium that contains fetal calf serum (FCS and growth factors cocktail for mass cultivation of human fibroblasts. To study proliferative activity of the medium on cultures of HUVEC cells of mesenchymal origin and mesenchymal stromal cells, as well as on cell culture of human amniotic fluid.Materials and methods. Determination of the rate of accumulation of the cellular mass and cell morphology in the course of cultivation of cells of various histogenesis in the Amniocar medium and nutrient medium that contains 10 % of FCS.Results. It has been demonstrated that the Amniocar medium is prevalent as compared to the standard DMEM medium with 10 % of FCS by 2 to 5 times for cultivation of skin fibroblasts, HUVEC, and mesenchymal stem cells. The Amniocar medium increased the quantity of endothelial cells that enter mitosis and maintained the culture of HUVEC cells with prolonged passaging in vitro. Clonal cultivation of human amniotic fluid cells in the Amniocar medium secured development of colonies of both fibroblast and epithelial type.Conclusions. Proliferative Amniocar medium is efficient for mass cultivation of various cells of mesenchymal origin and can be used for diagnostic purposes in medical genetics, oncology, etc.

  11. Radon in geological medium

    Energy Technology Data Exchange (ETDEWEB)

    Hricko, J [GEOCOMPLEX, a.s., Bratislava (Slovakia)

    1996-12-31

    The paper presented deals with behavior of the radon in geological medium and with some results of the radon survey in Bratislava and Kosice regions. 1) The a{sub v} has been detected in the holes 0.80 m deep. The density of observations - 3 reference areas (one represents 20 stations) per 1 km{sup 2}. The radon risk maps in 1:25000 and 1:50000 scales have been compiled. The 56.8% of the project area lies in low radon risk, 37.6% in medium radon risk and 5.6% in high radon risk. Follow-up monitoring of the equivalent volume radon activity (EVRA) at the flats, located in the areas with high radon risk of the surface layer, has showed values several times higher than Slovak limits (Marianka, Raca, Vajnory). The evidence that neotectonic is excellent medium for rising up emanation to the subsurface layer, is shown on the map. The tectonic zone of Liscie udolie in Bratislava-Karlova Ves area has been clearly detected by profile radon survey (a{sub v} > 50 kBq/m{sup 3}). 2) At present, northern half of the area of Kosice in question was covered by radon survey. The low and medium radon risks have been observed here, while localities with high radon risk are small in extent. The part of radon risk and soil permeability map from northern Kosice area is shown. (J.K.) 3 figs., 2 refs.

  12. Radon in geological medium

    International Nuclear Information System (INIS)

    Hricko, J.

    1995-01-01

    The paper presented deals with behavior of the radon in geological medium and with some results of the radon survey in Bratislava and Kosice regions. 1) The a v has been detected in the holes 0.80 m deep. The density of observations - 3 reference areas (one represents 20 stations) per 1 km 2 . The radon risk maps in 1:25000 and 1:50000 scales have been compiled. The 56.8% of the project area lies in low radon risk, 37.6% in medium radon risk and 5.6% in high radon risk. Follow-up monitoring of the equivalent volume radon activity (EVRA) at the flats, located in the areas with high radon risk of the surface layer, has showed values several times higher than Slovak limits (Marianka, Raca, Vajnory). The evidence that neotectonic is excellent medium for rising up emanation to the subsurface layer, is shown on the map. The tectonic zone of Liscie udolie in Bratislava-Karlova Ves area has been clearly detected by profile radon survey (a v > 50 kBq/m 3 ). 2) At present, northern half of the area of Kosice in question was covered by radon survey. The low and medium radon risks have been observed here, while localities with high radon risk are small in extent. The part of radon risk and soil permeability map from northern Kosice area is shown. (J.K.) 3 figs., 2 refs

  13. Intramammary infection with coagulase-negative staphylococci at parturition: Species-specific prevalence, risk factors, and effect on udder health.

    Science.gov (United States)

    De Visscher, A; Piepers, S; Haesebrouck, F; De Vliegher, S

    2016-08-01

    Coagulase-negative staphylococci (CNS) are the main cause of bovine intramammary infections (IMI) in many countries. Despite a high prevalence of CNS IMI at parturition, species-specific risk factor studies, relying on accurate identification methods, are lacking. Therefore, this observational study aimed at determining the prevalence and distribution of different CNS species causing IMI in fresh heifers and dairy cows in Flemish dairy herds and identifying associated species- and subgroup-specific risk factors at the herd, cow, and quarter level. The effect on udder health was investigated as well. Staphylococcus chromogenes, S. sciuri, and S. cohnii were the most frequently isolated species. The only CNS species causing IMI in fresh heifers and dairy cows in all herds was Staphylococcus chromogenes, whereas large between-herd differences in distribution were observed for the other species. Quarters from heifers and quarters with an inverted teat end had higher odds of being infected with S. chromogenes, S. simulans, or S. xylosus as well as with S. chromogenes solely. Prepartum teat apex colonization with S. chromogenes increased the likelihood of S. chromogenes IMI in the corresponding quarters at parturition. Quarters with dirty teat apices before calving were more likely to be infected with S. cohnii, S. equorum, S. saprophyticus, or S. sciuri, supporting the environmental nature of these CNS species. Three species (S. chromogenes, S. simulans, and S. xylosus) were associated with a higher quarter somatic cell count at parturition as compared with uninfected quarters. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  14. Evaluation of epidermal growth factor receptor (EGFR) by chromogenic in situ hybridization (CISH) and immunohistochemistry (IHC) in archival gliomas using bright-field microscopy.

    Science.gov (United States)

    Marquez, Abbey; Wu, Rina; Zhao, Jianxin; Tao, Jianhua; Shi, Zuorong

    2004-03-01

    Overexpression of EGFR secondary to EGFR gene amplification is a common feature in primary malignant gliomas. To correctly assess EGFR protein and gene level as possible prognostic and predictive markers in gliomas, straightforward assays, which can be used routinely in the pathology laboratory to evaluate EGFR status, becomes critical. EGFR gene amplification and chromosome 7 aneuploidy was detected in 34 formalin-fixed, paraffin-embedded benign and malignant gliomas by chromogenic in situ hybridization (CISH) using digoxigenin-labeled EGFR and biotin-labeled chromosome 7 centromeric probes. The results were evaluated by bright-field microscopy under a 40x objective lens. EGFR protein level was detected by immunohistochemistry (IHC) using monoclonal antibody 31G7. Five cases, 3 astrocytoma grade III (33%) and 2 glioblastoma multiforme (GBM) (33%), had EGFR amplification displayed as diaminobenzidine-stained multiple dots suggesting the pattern of double-minute chromosomes. Chromosome 7 polysomy was found in 68% gliomas, 100% GBM, 67% astrocytoma grade III, 42% astrocytoma grade II, 50% astrocytoma grade I, 100% ependymoma, and the 1 case of mixed glioma III. High expression of EGFR protein was present in 62% gliomas and displayed membrane and cytoplasmic staining. All tumors with EGFR gene amplification showed EGFR high expression. High expression of EGFR without gene amplification was observed in all grades of gliomas. Simultaneous detection of EGFR gene copies or chromosome 7 centromere signals along with tissue morphology allows us to compare CISH results easily with IHC results. Our results show that CISH is an objective, practical, and accurate assay to screen for EGFR gene status in gliomas.

  15. Physical processes in the interstellar medium

    CERN Document Server

    Spitzer, Lyman

    2008-01-01

    Physical Processes in the Interstellar Medium discusses the nature of interstellar matter, with a strong emphasis on basic physical principles, and summarizes the present state of knowledge about the interstellar medium by providing the latest observational data. Physics and chemistry of the interstellar medium are treated, with frequent references to observational results. The overall equilibrium and dynamical state of the interstellar gas are described, with discussions of explosions produced by star birth and star death and the initial phases of cloud collapse leading to star formation.

  16. Stochastic histories of dust grains in the interstellar medium

    International Nuclear Information System (INIS)

    Liffman, K.; Clayton, D.D.

    1989-01-01

    The purpose is to study an evolving system of refractory dust grains within the Interstellar Medium (ISM). This is done via a combination of Monte Carlo processes and a system of partial differential equations, where refractory dust grains formed within supernova remnants and ejecta from high mass loss stars are subjected to the processes of sputtering and collisional fragmentation in the diffuse media and accretion within the cold molecular clouds. In order to record chemical detail, the authors take each new particle to consist of a superrefractory core plus a more massive refractory mantle. The particles are allowed to transfer to and fro between the different phases of the interstellar medium (ISM) - on a time scale of 10(exp 8) years - until either the particles are destroyed or the program finishes at a Galaxy time of 6x10(exp 9) years. The resulting chemical and size spectrum(s) are then applied to various astrophysical problems with the following results. For an ISM which has no collisional fragmentation of the dust grains, roughly 10 percent by mass of the most refractory material survives the rigors of the ISM intact, which leaves open the possibility that fossilized isotopically anomalous material may have been present within the primordial solar nebula. Stuctured or layered refractory dust grains within the model cannot explain the observed interstellar depletions of refractory material. Fragmentation due to grain-grain collisions in the diffuse phase plus the accretion of material in the molecular cloud phase can under certain circumstances cause a bimodal distribution in grain size

  17. Chromogenic in situ hybridisation for the assessment of HER2 status in breast cancer: an international validation ring study

    Science.gov (United States)

    van de Vijver, Marc; Bilous, Michael; Hanna, Wedad; Hofmann, Manfred; Kristel, Petra; Penault-Llorca, Frédérique; Rüschoff, Josef

    2007-01-01

    Introduction Before any new methodology can be introduced into the routine diagnostic setting it must be technically validated against the established standards. To this end, a ring study involving five international pathology laboratories was initiated to validate chromogenic in situ hybridisation (CISH) against fluorescence in situ hybridisation (FISH) and immunohistochemistry (IHC) as a test for assessing human epidermal growth factor receptor 2 (HER2) status in breast cancer. Methods Each laboratory performed CISH, FISH and IHC on its own samples. Unstained sections from each case were also sent to another participating laboratory for blinded retesting by CISH ('outside CISH'). Results A total of 211 invasive breast carcinoma cases were tested. In 76 cases with high amplification (HER2/CEP17 ratio >4.0) by FISH, 73 cases (96%) scored positive (scores ≥ 6) by 'outside CISH'. For FISH-negative cases (HER2/CEP17 ratio CISH scores indicating no amplification (score ≤ 5), and only three cases were positive by CISH; in the three remaining cases, no CISH result could be obtained. For cases with low-level amplification using FISH (HER2/CEP17 ratio 2.0–4.0), 20 of 35 had CISH scores indicating gene amplification. Inter-laboratory concordance was also very high: 95% for normal HER2 copy number (1–5 copies); and 92% for cases with HER2 copy numbers ≥ 6. CISH intra-laboratory concordance with IHC was 92% for IHC-negative cases (IHC 0/1+) and 91% for IHC 3+ cases. Among IHC 2+ cases, CISH was 100% concordant with samples showing high amplification by FISH, and 94% concordant with FISH-negative samples. Conclusion These results show that CISH inter- and intra-laboratory concordance to FISH and IHC is very high, even in equivocal IHC 2+ cases. Therefore, we conclude that CISH is a methodology that is a viable alternative to FISH in the HER2 testing algorithm. PMID:17922920

  18. HER-2 gene amplification by chromogenic in situ hybridisation (CISH) compared with fluorescence in situ hybridisation (FISH) in breast cancer-A study of two hundred cases.

    Science.gov (United States)

    Sáez, A; Andreu, F J; Seguí, M A; Baré, M L; Fernández, S; Dinarés, C; Rey, M

    2006-08-01

    The purpose of the study was to compare two methods used to analyse HER-2 gene amplification (fluorescence in situ hybridisation (FISH) and chromogenic in situ hybridisation (CISH)), and determine the accuracy of the antibodies CB11 and HercepTest for immunohistochemical detection of HER-2 overexpression from archival breast cancer tissue. Additionally, interobserver variability in the interpretation of CISH and immunohistochemical tests was measured. Two hundred cases of invasive breast carcinoma diagnosed between 2000 and 2003 were selected. Immunohistochemistry (IHC) was performed with HercepTest and CB11, and gene amplification was determined by FISH (PathVision, Vysis) and CISH (Zymed) using tissue macroarrays. An excellent concordance (94.8%) was found between CISH and FISH. Considering FISH as gold standard, sensitivity of CISH was 97.5% and specificity 94%. Overall interobserver agreement of CISH was 97.5% and of IHC 84%. Both antibodies showed a sensitivity of 95.2% and a specificity of 70.7% (CB11) and 81.2% (HercepTest). Our results show that CISH is a highly accurate, reproducible and practical technique to determine HER-2 gene amplification. CB11 and HercepTest are good screening methods with a high sensitivity. The performance of tissue macroarrays to test HER-2 status by IHC, FISH and CISH has demonstrated to be an available and effective method to study large series of tumours.

  19. Effects of Nitric Oxide Production Inhibitor Named, NG-Nitro-L-Arginine Methyl Ester (L-NAME, on Rat Mesenchymal Stem Cells Differentiation

    Directory of Open Access Journals (Sweden)

    E Arfaei

    2010-04-01

    Full Text Available Introduction & Objectives: Recently, the findings of some studies have shown that, nitric oxide (NO probably has an important role in differentiation of mesenchymal stem cells to osteoblasts. The aim of the present investigation was to study the effects of nitric oxide production inhibitor named, NG-nitro-L-arginine methyl ester (L-NAME, on rat mesenchymal stem cells differentiation to osteoblasts in vitro. Materials & Methods: This was an experimental study conducted at Hamedan University of Medical Sciences in 2009, in which rat bone marrow stem cells were isolated in an aseptic condition and cultured in vitro. After third passage, the cells were cultured in osteogenic differentiation medium. To study the effects of L-NAME on osteogenic differentiation, the L-NAME was added to the culture medium at a concentration of 125, 250, and 500 μM in some culture plates. During the culture procedure, the media were replaced with fresh ones, with a three days interval. After 28 days of culturing the mineralized matrix was stained using Alizarian red staining method. The gathered data were analyzed by SPSS software version 12 using one way ANOVA. Results: The findings of this study showed that in the presence of L-NAME, differentiation of bone marrow mesenchymal stem cells to osteoblasts was disordered and matrix mineralization significantly decreased in a dose dependent manner. Conclusion: This study revealed that, inhibition of nitric oxide production using L-NAME can prevent the differentiation of rat bone marrow mesenchymal stem cells to osteoblast. The results imply that NO is an important constituent in differentiation of mesenchymal stem cell to osteoblasts.

  20. The effect of hydrostatic pressure on staurosporine-induced neural differentiation in mouse bone marrow‑derived mesenchymal stem cells.

    Science.gov (United States)

    Javanmard, F; Azadbakht, M; Pourmoradi, M

    2016-01-01

    In this study, the role of hydrostatic pressure on staurosporine-induced neural differentiation in mouse bone marrow mesenchymal stem cells were investigated. The cells were cultured in treatment medium containing 100 nM of staurosporine for 4 hours; then the cells were affected by hydrostatic pressure (0, 25,50, 100 mmHg). The percentage of cell viability by trypan blue staining and the percentage of cell death by Hoechst/PI differential staining were assessed. We obtained the total neurite length. Expression of β-tubulin III and GFAP (Glial fibrillary acidic protein) proteins were also analyzed by immunocytochemistry. The percentage of cell viability in treatments decreased relative to the increase in hydrostatic pressure and time (p Keywords: bone marrow mesenchymal stem cell, hydrostatic pressure, immunocytochemistry, neural differentiation, neurite length, cell differentiation.

  1. Analysis of Epstein Barr Virus Encoded RNA Expression in Nasopharyngeal Carcinoma in North-Eastern India: A Chromogenic in Situ Hybridization Based Study

    Directory of Open Access Journals (Sweden)

    Anjan Saikia

    2016-05-01

    Full Text Available Introduction: Nasopharyngeal carcinoma (NPC is a common cancer in the North-East region of India. Though the role of environmental contributors of NPC in the North-Eastern part of India is firmly established, EBV as an etiological agent in the region remains unexplored. Material and Methods: Fifty-one patients, who presented at the department of ENT, NEIGRIHMS and were confirmed as NPC upon histopathological examination, were included in the study. Chromogenic in-situ hybridization (CISH was used for the evaluation of EBER (Epstein Barr Virus Encoded RNA. Presence of nuclear signals was taken as positive for EBER expression. EBER status was correlated with various clinicopathological parameters like age, sex, dietary habits, histological types of NPC, and ethnicity of the patients. Results: The age range of the study group was 25 to 70 years with a mean age of 44.64 years and a male:female ratio of 3:2. Non-keratinizing undifferentiated type of NPC was the most common histological type. EBV was positive in 59% (30/51 of our cases. It showed a statistically significant correlation with the Naga community (P=0.01, with consumption of smoked food (P=0.02, and cigarette smoking (P=0.02. There was no correlation of EBV with age, sex, lymph node metastasis, stage, and histology. Conclusion: Our result indicates that EBV may be an additional risk factor in the pathogenesis of NPC in this region of India. So apart from lifestyle modification, a future study for a screening test for EBV viral load even in asymptomatic patients may be considered, for determination of disease susceptibility, early diagnosis, and proper management.

  2. Comparison of Clot-based, Chromogenic, and Fluorescence Assays for Measurement of Factor VIII Inhibitors in the U.S. Hemophilia Inhibitor Research Study

    Science.gov (United States)

    Miller, Connie H.; Rice, Anne S.; Boylan, Brian; Shapiro, Amy D.; Lentz, Steven R.; Wicklund, Brian M.; Kelly, Fiona M.; Soucie, J. Michael

    2015-01-01

    Summary Background Detection and validation of inhibitors (antibodies) to hemophilia treatment products are important for clinical care, evaluation of product safety, and assessment of population trends. Methods Centralized monitoring for factor VIII (FVIII) inhibitors was conducted for patients in the Hemophilia Inhibitor Research Study using a previously reported modified Nijmegen-Bethesda clotting assay (NBA), a chromogenic Bethesda assay (CBA), and a novel fluorescence immunoassay (FLI). Results NBA and CBA were performed on 1005 specimens and FLI on 272 specimens. CBA was negative on 880/883 specimens (99.7%) with Nijmegen-Bethesda units (NBU)NBA and negative CBA, 58.1% were FLI-negative, 12.9% had evidence of lupus anticoagulant, and 35.5% had non-time-dependent inhibition. CBA and FLI were positive on 72.4% and 100% of 1.0–1.9 NBU specimens and 43.1% and 50.0% of 0.5–0.9 NBU specimens. FLI detected antibodies in 98.0% of CBA-positive and 81.6% of NBA-positive specimens (P=0.004). Among 21 new inhibitors detected by NBA, 5 (23.8%) with 0.7–1.3 NBU did not react in CBA or FLI. Among previously positive patients with 0.5–1.9 NBU, 7/25 (28%) were not CBA or FLI positive. FLI was positive on 36/169 NBU-negative specimens (21.3%). Conclusions FVIII specificity could not be demonstrated by CBA or FLI for 26% of inhibitors of 0.5–1.9 NBU; such results must be interpreted with caution. Low titer inhibitors detected in clot-based assays should always be repeated, with consideration given to evaluating their reactivity with FVIII using more specific assays. PMID:23601690

  3. Osteocytes subjected to pulsating fluid flow regulate osteoblast proliferation and differentiation

    International Nuclear Information System (INIS)

    Vezeridis, Peter S.; Semeins, Cornelis M.; Chen Qian; Klein-Nulend, Jenneke

    2006-01-01

    Osteocytes are thought to orchestrate bone remodeling, but it is unclear exactly how osteocytes influence neighboring bone cells. Here, we tested whether osteocytes, osteoblasts, and periosteal fibroblasts subjected to pulsating fluid flow (PFF) produce soluble factors that modulate the proliferation and differentiation of cultured osteoblasts and periosteal fibroblasts. We found that osteocyte PFF conditioned medium (CM) inhibited bone cell proliferation, and osteocytes produced the strongest inhibition of proliferation compared to osteoblasts and periosteal fibroblasts. The nitric oxide (NO) synthase inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME) attenuated the inhibitory effects of osteocyte PFF CM, suggesting that a change in NO release is at least partially responsible for the inhibitory effects of osteocyte PFF CM. Furthermore, osteocyte PFF CM stimulated osteoblast differentiation measured as increased alkaline phosphatase activity, and L-NAME decreased the stimulatory effects of osteocyte PFF CM on osteoblast differentiation. We conclude that osteocytes subjected to PFF inhibit proliferation but stimulate differentiation of osteoblasts in vitro via soluble factors and that the release of these soluble factors was at least partially dependent on the activation of a NO pathway in osteocytes in response to PFF. Thus, the osteocyte appears to be more responsive to PFF than the osteoblast or periosteal fibroblast with respect to the production of soluble signaling molecules affecting osteoblast proliferation and differentiation

  4. Bufalin inhibits the differentiation and proliferation of human osteosarcoma cell line hMG63-derived cancer stem cells.

    Science.gov (United States)

    Chang, Yuewen; Zhao, Yongfang; Zhan, Hongsheng; Wei, Xiaoen; Liu, Tianjin; Zheng, Bo

    2014-02-01

    Cancer stem cells (CSCs) play an important role in drug resistance of tumor and are responsible for high recurrence rates. Agents that can suppress the proliferation and differentiation of CSCs would provide new opportunity to fight against tumor recurrence. In this study, we developed a new strategy to enrich CSCs in human osteosarcoma cell line hMG63. Using these CSCs as model, we tested the effect of bufalin, a traditional Chinese medicine, on the proliferation and differentiation of CSCs. hMG63 cells were cultured in poly-HEMA-treated dish and cancer stem cell-specific medium. In this nonadhesive culture system, hMG63 formed spheres, which were then collected and injected into the immunodeficient mice. Cisplatin was administered every 3 days for five times. The enriched xenograft tumors were cultured in cancer stem cell-specific medium again to form tumor spheres. Expression of cancer stem cell markers of these cells was measured by flow cytometry. These cells were then treated with bufalin, and the proliferation and differentiation ability were indicated by the expression level of molecular markers and the formation of sphere again in vitro. We obtained a low CD133+/CD44 cell population with high-level stem cell marker. When treated with bufalin, the sphere could not get attached to the flask and failed to differentiate, which was indicated by the stable expression of stem cell marker CD133 and OCT-4 in the condition permissive to differentiation. Treatment of bufalin also suppressed the single cells isolated from the sphere to form sphere again in the nonadhesive culture system, and a decreased expression of proliferation marker Ki67 was also detected in these cells. Sphere-formed and chemoresistant colon xenograft tumors in immunodeficient mice could enrich cancer stem cell population. Bufalin could inhibit proliferation and differentiation of CSCs.

  5. The differentiation potential of adipose tissue-derived mesenchymal stem cells into cell lineage related to male germ cells

    Directory of Open Access Journals (Sweden)

    P. Bräunig

    Full Text Available ABSTRACT The adipose tissue is a reliable source of Mesenchymal stem cells (MSCs showing a higher plasticity and transdifferentiation potential into multilineage cells. In the present study, adipose tissue-derived mesenchymal stem cells (AT-MSCs were isolated from mice omentum and epididymis fat depots. The AT-MSCs were initially compared based on stem cell surface markers and on the mesodermal trilineage differentiation potential. Additionally, AT-MSCs, from both sources, were cultured with differentiation media containing retinoic acid (RA and/or testicular cell-conditioned medium (TCC. The AT-MSCs expressed mesenchymal surface markers and differentiated into adipogenic, chondrogenic and osteogenic lineages. Only omentum-derived AT-MSCs expressed one important gene marker related to male germ cell lineages, after the differentiation treatment with RA. These findings reaffirm the importance of adipose tissue as a source of multipotent stromal-stem cells, as well as, MSCs source regarding differentiation purpose.

  6. Effective medium theory for anisotropic metamaterials

    KAUST Repository

    Zhang, Xiujuan; Wu, Ying

    2015-01-01

    -dimensional metamaterial composed of a rectangular array of elliptic cylinders and derive an effective medium theory for such a metamaterial. We find that it is possible to obtain a closed-form analytical solution for the anisotropic effective medium parameters, provided

  7. The tyrosine kinase inhibitor dasatinib induces a marked adipogenic differentiation of human multipotent mesenchymal stromal cells.

    Directory of Open Access Journals (Sweden)

    Adriana Borriello

    Full Text Available BACKGROUND: The introduction of specific BCR-ABL inhibitors in chronic myelogenous leukemia therapy has entirely mutated the prognosis of this hematologic cancer from being a fatal disorder to becoming a chronic disease. Due to the probable long lasting treatment with tyrosine-kinase inhibitors (TKIs, the knowledge of their effects on normal cells is of pivotal importance. DESIGN AND METHODS: We investigated the effects of dasatinib treatment on human bone marrow-derived mesenchymal stromal cells (MSCs. RESULTS: Our findings demonstrate, for the first time, that dasatinib induces MSCs adipocytic differentiation. Particularly, when the TKI is added to the medium inducing osteogenic differentiation, a high MSCs percentage acquires adipocytic morphology and overexpresses adipocytic specific genes, including PPARγ, CEBPα, LPL and SREBP1c. Dasatinib also inhibits the activity of alkaline phosphatase, an osteogenic marker, and remarkably reduces matrix mineralization. The increase of PPARγ is also confirmed at protein level. The component of osteogenic medium required for dasatinib-induced adipogenesis is dexamethasone. Intriguingly, the increase of adipocytic markers is also observed in MSCs treated with dasatinib alone. The TKI effect is phenotype-specific, since fibroblasts do not undergo adipocytic differentiation or PPARγ increase. CONCLUSIONS: Our data demonstrate that dasatinib treatment affects bone marrow MSCs commitment and suggest that TKIs therapy might modify normal phenotypes with potential significant negative consequences.

  8. Characterization of Chondrogenic Gene Expression and Cartilage Phenotype Differentiation in Human Breast Adipose-Derived Stem Cells Promoted by Ginsenoside Rg1 In Vitro

    Directory of Open Access Journals (Sweden)

    Fang-Tian Xu

    2015-11-01

    Full Text Available Background/Aims: Investigating and understanding chondrogenic gene expression during the differentiation of human breast adipose-derived stem cells (HBASCs into chondrogenic cells is a prerequisite for the application of this approach for cartilage repair and regeneration. In this study, we aim to characterize HBASCs and to examine chondrogenic gene expression in chondrogenic inductive culture medium containing ginsenoside Rg1. Methods: Human breast adipose-derived stem cells at passage 3 were evaluated based on specific cell markers and their multilineage differentiation capacity. Cultured HBASCs were treated either with basic chondrogenic inductive conditioned medium alone (group A, control or with basic chondrogenic inductive medium plus 10 µg/ml (group B, 50 µg/ml (group C, or 100µg/ml ginsenoside Rg1 (group D. Cell proliferation was assessed using the CCK-8 assay for a period of 9 days. Two weeks after induction, the expression of chondrogenic genes (collagen type II, collagen type XI, ACP, COMP and ELASTIN was determined using real-time PCR in all groups. Results: The different concentrations of ginsenoside Rg1 that were added to the basic chondrogenic inductive culture medium promoted the proliferation of HBASCs at earlier stages (groups B, C, and D but resulted in chondrogenic phenotype differentiation and higher mRNA expression of collagen type II (CO-II, collagen type XI (CO-XI, acid phosphatase (ACP, cartilage oligomeric matrix protein (COMP and ELASTIN compared with the control (group A at later stages. The results reveal an obvious positive dose-effect relationship between ginsenoside Rg1 and the proliferation and chondrogenic phenotype differentiation of HBASCs in vitro. Conclusions: Human breast adipose-derived stem cells retain stem cell characteristics after expansion in culture through passage 3 and serve as a feasible source of cells for cartilage regeneration in vitro. Chondrogenesis in HBASCs was found to be prominent

  9. Improved lipid and biomass productivities in Chlorella vulgaris by differing the inoculation medium from the production medium

    Directory of Open Access Journals (Sweden)

    Shahrbanoo Hamedi

    2016-06-01

    Full Text Available Improvement of biomass and lipid productivities is now one of the main concerns in commercialization of microalgae cultivation as a feedstock for algal biofuel production. Conventional photoautotrophic processes using well-studied and rich in oil strain of Chlorella vulgaris are not able to meet such demands. A new strategy of inoculating algae production medium with cells grown in a different medium from the production medium was proposed herein. More specifically, when SH4 was used as production medium and N8 was used as inoculation medium, biomass and lipid productivities increased by 2.33 folds and 1.44 fold, respectively, compared with when the production and inoculation media were the same, such as SH4. The findings of the present investigation showed that this cultivation scheme resulted in 52% increase in cell number and 54% increase in dry weight leading to improved productivities. Although by even considering this improvement, photoautotrophic cultivation of algae can hardly compete with the heterotrophic cultivation, the high cost of hydrocarbon supply required in large-scale heterotrophic processes marks the technique proposed in the present study as a promising approach for commercialization of algal biofuel production.

  10. Effect of essential amino acids on enteroids: Methionine deprivation suppresses proliferation and affects differentiation in enteroid stem cells

    International Nuclear Information System (INIS)

    Saito, Yuki; Iwatsuki, Ken; Hanyu, Hikaru; Maruyama, Natsuki; Aihara, Eitaro; Tadaishi, Miki; Shimizu, Makoto; Kobayashi-Hattori, Kazuo

    2017-01-01

    We investigated the effects of essential amino acids on intestinal stem cell proliferation and differentiation using murine small intestinal organoids (enteroids) from the jejunum. By selectively removing individual essential amino acids from culture medium, we found that 24 h of methionine (Met) deprivation markedly suppressed cell proliferation in enteroids. This effect was rescued when enteroids cultured in Met deprivation media for 12 h were transferred to complete medium, suggesting that Met plays an important role in enteroid cell proliferation. In addition, mRNA levels of the stem cell marker leucine-rich repeat-containing G protein-coupled receptor 5 (Lgr5) decreased in enteroids grown in Met deprivation conditions. Consistent with this observation, Met deprivation also attenuated Lgr5-EGFP fluorescence intensity in enteroids. In contrast, Met deprivation enhanced mRNA levels of the enteroendocrine cell marker chromogranin A (ChgA) and markers of K cells, enterochromaffin cells, goblet cells, and Paneth cells. Immunofluorescence experiments demonstrated that Met deprivation led to an increase in the number of ChgA-positive cells. These results suggest that Met deprivation suppresses stem cell proliferation, thereby promoting differentiation. In conclusion, Met is an important nutrient in the maintenance of intestinal stem cells and Met deprivation potentially affects cell differentiation. - Highlights: • Met influences the proliferation of enteroids. • Met plays a crucial role in the maintenance of stem cells. • Met deprivation potentially promotes differentiation into secretory cells.

  11. The influence of scaffold microstructure on chondrogenic differentiation of mesenchymal stem cells

    International Nuclear Information System (INIS)

    Zhang, Jingjing; Ge, Zigang; Wu, Yingnan; Lee, Eng Hin; Thote, Tanushree; Yang, Zheng

    2014-01-01

    Different forms of biomaterials, including microspheres, sponges, hydrogels and nanofibres have been broadly used in cartilage regeneration; however, effects of internal structures of biomaterials on chondrogenesis of mesenchymal stem cells (MSCs) remain largely unexplored. Here we investigated the effect of physical microenvironments of sponges and hydrogels on chondrogenic differentiation of MSCs. MSCs, cultured in these two scaffold systems, were induced with TGF-β 3  in chondrogeneic differentiation medium and the chondrogenic differentiation was evaluated and compared after three weeks. MSCs in the sponges clustered with spindle morphologies, while they distributed homogenously with round morphologies in the hydrogel. The MSCs proliferated faster in the sponge compared to that in the hydrogel. Significantly higher glycosaminoglycan and collagen II were found in the sponges but not in the hydrogels. The different tissue formation ability of MSCs in these two systems could be attributed to the different metabolic requirements and the cellular events prerequisite in the chondrogenic process of MSCs. It is reasonable to conclude that sponges with relatively active microenvironments that facilitate cell–cell contacts and cell–matrix interaction are optimal for early stage of chondrogeneic differentiation. (paper)

  12. Glycosaminoglycan-Mimetic Signals Direct the Osteo/Chondrogenic Differentiation of Mesenchymal Stem Cells in a Three-Dimensional Peptide Nanofiber Extracellular Matrix Mimetic Environment.

    Science.gov (United States)

    Arslan, Elif; Guler, Mustafa O; Tekinay, Ayse B

    2016-04-11

    Recent efforts in bioactive scaffold development focus strongly on the elucidation of complex cellular responses through the use of synthetic systems. Designing synthetic extracellular matrix (ECM) materials must be based on understanding of cellular behaviors upon interaction with natural and artificial scaffolds. Hence, due to their ability to mimic both the biochemical and mechanical properties of the native tissue environment, supramolecular assemblies of bioactive peptide nanostructures are especially promising for development of bioactive ECM-mimetic scaffolds. In this study, we used glycosaminoglycan (GAG) mimetic peptide nanofiber gel as a three-dimensional (3D) platform to investigate how cell lineage commitment is altered by external factors. We observed that amount of fetal bovine serum (FBS) presented in the cell media had synergistic effects on the ability of GAG-mimetic nanofiber gel to mediate the differentiation of mesenchymal stem cells into osteogenic and chondrogenic lineages. In particular, lower FBS concentration in the culture medium was observed to enhance osteogenic differentiation while higher amount FBS promotes chondrogenic differentiation in tandem with the effects of the GAG-mimetic 3D peptide nanofiber network, even in the absence of externally administered growth factors. We therefore demonstrate that mesenchymal stem cell differentiation can be specifically controlled by the combined influence of growth medium components and a 3D peptide nanofiber environment.

  13. Study of niobium corrosion in alkaline medium

    International Nuclear Information System (INIS)

    Almeida, S.H. de.

    1987-01-01

    A comparative study of niobium electrochemical behaviour in NaOH and KOH solution, with concentrations between 0,5 and 6,1M is presented. The studies were done through electrochemicals assays, consisting in the corrosion potential and anodic and cathodic polarization curves, complemented by loss of mass experiments. The niobium anodic behaviour in alkaline medium is characterized by passivation occurrence, with a stable film formation. The Na oH solution in alkaline medium are more corrosible to niobium than the KOH solution. The loss of mass assays showed that the corrosion velocit is more dependente of hydroxide concentration in KOH medium than the NaOH medium. (C.G.C.) [pt

  14. Somatic cell count and milk neutrophil viability of dairy heifers with specific CXCR1 genotypes following experimental intramammary infection with Staphylococcus chromogenes originating from milk.

    Science.gov (United States)

    Verbeke, Joren; Piccart, Kristine; Piepers, Sofie; Van Poucke, Mario; Peelman, Luc; De Visscher, Anneleen; De Vliegher, Sarne

    2015-06-01

    Previous observational studies suggest an association between polymorphism c.980A>G in the CXCR1 gene, encoding the chemokine (C-X-C motif) receptor 1, and the innate immunity and infection status of the mammary gland. Mammary glands of eight Holstein heifers were experimentally infected with a Staphylococcus chromogenes isolate originating from a chronic intramammary infection (IMI) to study differences between CXCR1 genotypes c.980AG and c.980GG. Quarters from heifers with genotypes c.980AG and c.980GG developed subclinical mastitis but showed differences in the early response at 6-18 h post challenge. Bacterial count at 18 h post challenge tended to be higher in quarters from c.980AG heifers compared to c.980GG heifers. Somatic cell count (SCC) was higher at 6 h post challenge and tended to be higher at 9 h post challenge in c.980AG heifers compared to c.980GG heifers. Milk production decreased similarly. Milk neutrophils of c.980AG heifers showed more apoptosis at 9 h post challenge and tended to show more necrosis at 6, 9 and 12 h post challenge than c.980GG heifers. Differences were less pronounced in the later stage (>18 h) of infection. The results demonstrate that CXCR1 polymorphism can influence SCC and milk neutrophil viability following experimental IMI. Copyright © 2015 Elsevier Ltd. All rights reserved.

  15. Pulsed DC Electric Field-Induced Differentiation of Cortical Neural Precursor Cells.

    Directory of Open Access Journals (Sweden)

    Hui-Fang Chang

    Full Text Available We report the differentiation of neural stem and progenitor cells solely induced by direct current (DC pulses stimulation. Neural stem and progenitor cells in the adult mammalian brain are promising candidates for the development of therapeutic neuroregeneration strategies. The differentiation of neural stem and progenitor cells depends on various in vivo environmental factors, such as nerve growth factor and endogenous EF. In this study, we demonstrated that the morphologic and phenotypic changes of mouse neural stem and progenitor cells (mNPCs could be induced solely by exposure to square-wave DC pulses (magnitude 300 mV/mm at frequency of 100-Hz. The DC pulse stimulation was conducted for 48 h, and the morphologic changes of mNPCs were monitored continuously. The length of primary processes and the amount of branching significantly increased after stimulation by DC pulses for 48 h. After DC pulse treatment, the mNPCs differentiated into neurons, astrocytes, and oligodendrocytes simultaneously in stem cell maintenance medium. Our results suggest that simple DC pulse treatment could control the fate of NPCs. With further studies, DC pulses may be applied to manipulate NPC differentiation and may be used for the development of therapeutic strategies that employ NPCs to treat nervous system disorders.

  16. Pulsed DC Electric Field-Induced Differentiation of Cortical Neural Precursor Cells.

    Science.gov (United States)

    Chang, Hui-Fang; Lee, Ying-Shan; Tang, Tang K; Cheng, Ji-Yen

    2016-01-01

    We report the differentiation of neural stem and progenitor cells solely induced by direct current (DC) pulses stimulation. Neural stem and progenitor cells in the adult mammalian brain are promising candidates for the development of therapeutic neuroregeneration strategies. The differentiation of neural stem and progenitor cells depends on various in vivo environmental factors, such as nerve growth factor and endogenous EF. In this study, we demonstrated that the morphologic and phenotypic changes of mouse neural stem and progenitor cells (mNPCs) could be induced solely by exposure to square-wave DC pulses (magnitude 300 mV/mm at frequency of 100-Hz). The DC pulse stimulation was conducted for 48 h, and the morphologic changes of mNPCs were monitored continuously. The length of primary processes and the amount of branching significantly increased after stimulation by DC pulses for 48 h. After DC pulse treatment, the mNPCs differentiated into neurons, astrocytes, and oligodendrocytes simultaneously in stem cell maintenance medium. Our results suggest that simple DC pulse treatment could control the fate of NPCs. With further studies, DC pulses may be applied to manipulate NPC differentiation and may be used for the development of therapeutic strategies that employ NPCs to treat nervous system disorders.

  17. Generation of megakaryocytic progenitors from human embryonic stem cells in a feeder- and serum-free medium.

    Directory of Open Access Journals (Sweden)

    Marjorie Pick

    Full Text Available BACKGROUND: The production of human platelets from embryonic stem cells in a defined culture system is a prerequisite for the generation of platelets for therapeutic use. As an important step towards this goal, we report the differentiation of human embryonic stem cells (hESCs towards the megakaryocyte (Mk lineage using a 'spin embryoid body' method in serum-free differentiation medium. METHODOLOGY AND PRINCIPAL FINDINGS: Immunophenotypic analyses of differentiating hESC identified a subpopulation of cells expressing high levels of CD41a that expressed other markers associated with the Mk lineage, including CD110, CD42b and CD61. Differentiated cells were sorted on the basis of their expression of CD41a, CD34 and CD45 and assessed for Mk colony formation, expression of myeloid and Mk genes and ability to endoreplicate DNA. In a collagen-based colony assay, the CD41a⁺ cells sorted from these differentiation cultures produced 100-800 Mk progenitors at day 13 and 25-160 Mk progenitors at day 20 of differentiation per 100,000 cells assayed. Differentiated Mk cells produced platelet-like particles which expressed CD42b and were activated by ADP, similar to platelets generated from precursors in cord blood. These studies were complemented by real time PCR analyses showing that subsets of cells enriched for CD41a⁺ Mk precursors expressed high levels of Mk associated genes such as PF4 and MPL. Conversely, high levels of myeloid and erythroid related transcripts, such as GATA1, TAL1/SCL and PU.1, were detected in sorted fractions containing CD34⁺ and CD45⁺ cells. CONCLUSIONS: We describe a serum- and feeder-free culture system that enabled the generation of Mk progenitors from human embryonic stem cells. These cells formed colonies that included differentiated Mks that fragmented to form platelet-like particles. This protocol represents an important step towards the generation of human platelets for therapeutic use.

  18. In vitro germ cell differentiation from cynomolgus monkey embryonic stem cells.

    Directory of Open Access Journals (Sweden)

    Kaori Yamauchi

    Full Text Available BACKGROUND: Mouse embryonic stem (ES cells can differentiate into female and male germ cells in vitro. Primate ES cells can also differentiate into immature germ cells in vitro. However, little is known about the differentiation markers and culture conditions for in vitro germ cell differentiation from ES cells in primates. Monkey ES cells are thus considered to be a useful model to study primate gametogenesis in vitro. Therefore, in order to obtain further information on germ cell differentiation from primate ES cells, this study examined the ability of cynomolgus monkey ES cells to differentiate into germ cells in vitro. METHODS AND FINDINGS: To explore the differentiation markers for detecting germ cells differentiated from ES cells, the expression of various germ cell marker genes was examined in tissues and ES cells of the cynomolgus monkey (Macaca fascicularis. VASA is a valuable gene for the detection of germ cells differentiated from ES cells. An increase of VASA expression was observed when differentiation was induced in ES cells via embryoid body (EB formation. In addition, the expression of other germ cell markers, such as NANOS and PIWIL1 genes, was also up-regulated as the EB differentiation progressed. Immunocytochemistry identified the cells expressing stage-specific embryonic antigen (SSEA 1, OCT-4, and VASA proteins in the EBs. These cells were detected in the peripheral region of the EBs as specific cell populations, such as SSEA1-positive, OCT-4-positive cells, OCT-4-positive, VASA-positive cells, and OCT-4-negative, VASA-positive cells. Thereafter, the effect of mouse gonadal cell-conditioned medium and growth factors on germ cell differentiation from monkey ES cells was examined, and this revealed that the addition of BMP4 to differentiating ES cells increased the expression of SCP1, a meiotic marker gene. CONCLUSION: VASA is a valuable gene for the detection of germ cells differentiated from ES cells in monkeys, and the

  19. Enhanced Chondrogenic Differentiation of Human Umbilical Cord Wharton's Jelly Derived Mesenchymal Stem Cells by GSK-3 Inhibitors.

    Directory of Open Access Journals (Sweden)

    Prapot Tanthaisong

    Full Text Available Articular cartilage is an avascular, alymphatic, and aneural system with very low regeneration potential because of its limited capacity for self-repair. Mesenchymal stem cells (MSCs are the preferred choice for cell-based therapies. Glycogen synthase kinase 3 (GSK-3 inhibitors are compounds that can induce the Wnt signaling pathway, which is involved in chondrogenesis and cartilage development. Here, we investigated the influence of lithium chloride (LiCl and SB216763 synergistically with TGF-β3 on chondrogenic differentiation in human mesenchymal stem cells derived from Wharton's jelly tissue (hWJ-MSCs. hWJ-MSCs were cultured and chondrogenic differentiation was induced in monolayer and pellet experiments using chondrogenic medium, chondrogenic medium supplemented with LiCl, or SB216763 for 4 weeks. After in vitro differentiation, cultured cells were examined for the expression of Sox9, ACAN, Col2a1, and β-catenin markers. Glycosaminoglycan (GAG accumulation was also examined by Alcian blue staining. The results indicated that SB216763 was more effective than LiCl as evidenced by a higher up-regulation of the expression of cartilage-specific markers, including Sox9, ACAN, Col2a1 as well as GAG accumulation. Moreover, collagen type II expression was strongly observed in cells cultured in the chondrogenic medium + SB216763 as evidenced by western blot analysis. Both treatments appeared to mediate the Wnt signaling pathway by up-regulating β-catenin gene expression. Further analyses showed that all treatments suppressed the progression of chondrocyte hypertrophy, determined by decreased expression of Col10a1 and Runx2. These results indicate that LiCl and SB216763 are potential candidates for further in vivo therapeutic trials and would be of great importance for cartilage regeneration.

  20. Adaptation of Listeria monocytogenes in a simulated cheese medium: effects on virulence using the Galleria mellonella infection model.

    Science.gov (United States)

    Schrama, D; Helliwell, N; Neto, L; Faleiro, M L

    2013-06-01

    The aim of this study was to evaluate the effect of the acid and salt adaptation in a cheese-based medium on the virulence potential of Listeria monocytogenes strains isolated from cheese and dairy processing environment using the Galleria mellonella model. Four L. monocytogenes strains were exposed to a cheese-based medium in conditions of induction of an acid tolerance response and osmotolerance response (pH 5·5 and 3·5% w/v NaCl) and injected in G. mellonella insects. The survival of insects and the L. monocytogenes growth kinetics in insects were evaluated. The gene expression of hly, actA and inlA genes was determined by real-time PCR. The adapted cells of two dairy strains showed reduced insect mortality (P 0·05) was found between adapted and nonadapted cells. The gene expression results evidenced an overexpression of virulence genes in cheese-based medium, but not in simulated insect-induced conditions. Our results suggest that adaptation to low pH and salt in a cheese-based medium can affect the virulence of L. monocytogenes, but this effect is strain dependent. In this study, the impact of adaptation to low pH and salt in a cheese-based medium on L. monocytogenes virulence was tested using the Wax Moth G. mellonella model. This model allowed the differentiation of the virulence potential between the L. monocytogenes strains. The effect of adaptation on virulence is strain dependent. The G. mellonella model revealed to be a prompt method to test food-related factors on L. monocytogenes virulence. © 2013 The Society for Applied Microbiology.

  1. Effects and mechanisms of melatonin on neural differentiation of induced pluripotent stem cells.

    Science.gov (United States)

    Shu, Tao; Wu, Tao; Pang, Mao; Liu, Chang; Wang, Xuan; Wang, Juan; Liu, Bin; Rong, Limin

    2016-06-03

    Melatonin, a lipophilic molecule mainly synthesized in the pineal gland, has properties of antioxidation, anti-inflammation, and antiapoptosis to improve neuroprotective functions. Here, we investigate effects and mechanisms of melatonin on neural differentiation of induced pluripotent stem cells (iPSCs). iPSCs were induced into neural stem cells (NSCs), then further differentiated into neurons in medium with or without melatonin, melatonin receptor antagonist (Luzindole) or Phosphatidylinositide 3 kinase (PI3K) inhibitor (LY294002). Melatonin significantly promoted the number of neurospheres and cell viability. In addition, Melatonin markedly up-regulated gene and protein expression of Nestin and MAP2. However, Luzindole or LY294002 attenuated these increase. The expression of pAKT/AKT were increased by Melatonin, while Luzindole or LY294002 declined these melatonin-induced increase. These results suggest that melatonin significantly increased neural differentiation of iPSCs via activating PI3K/AKT signaling pathway through melatonin receptor. Copyright © 2016 Elsevier Inc. All rights reserved.

  2. Small and medium power reactors 1987

    International Nuclear Information System (INIS)

    1987-12-01

    This TECDOC follows the publication of TECDOC-347 Small and Medium Power Reactors Project Initiation Study - Phase I published in 1985 and TECDOC-376 Small and Medium Power Reactors 1985 published in 1986. It is mainly intended for decision makers in Developing Member States interested in embarking on a nuclear power programme. It consists of two parts: 1) Guidelines for the Introduction of Small and Medium Power Reactors in Developing Countries. These Guidelines were established during the Advisory Group Meeting held in Vienna from 11 to 15 May 1987. Their purpose is to review key aspects relating to the introduction of Small and Medium Power Reactors in developing countries; 2) Up-dated Information on SMPR Concepts Contributed by Supplier Industries. According to the recommendations of the Second Technical Committee Meeting on SMPRs held in Vienna in March 1985, this part contains the up-dated information formerly published in Annex I of the above mentioned TECDOC-347. Figs

  3. Effective medium theory principles and applications

    CERN Document Server

    Choy, Tuck C

    2015-01-01

    Effective medium theory dates back to the early days of the theory of electricity. Faraday in 1837 proposed one of the earliest models for a composite metal-insulator dielectric and around 1870 Maxwell and later Garnett (1904) developed models to describe a composite or mixed material medium. The subject has been developed considerably since and while the results are useful for predicting materials performance, the theory can also be used in a wide range of problems in physics and materials engineering. This book develops the topic of effective medium theory by bringing together the essentials of both the static and the dynamical theory. Electromagnetic systems are thoroughly dealt with, as well as related areas such as the CPA theory of alloys, liquids, the density functional theory etc., with applications to ultrasonics, hydrodynamics, superconductors, porous media and others, where the unifying aspects of the effective medium concept are emphasized. In this new second edition two further chapters have been...

  4. Small and medium power reactors 1987

    Science.gov (United States)

    1987-12-01

    This TECDOC follows the publication of TECDOC-347: Small and Medium Power Reactors (SMPR) Project Initiation Study, Phase 1, published in 1985 and TECDOC-376: Small and Medium Power Reactors 1985 published in 1986. It is mainly intended for decision makers in Developing Member States interested in embarking on a nuclear power program. It consists of two parts: (1) guidelines for the introduction of small and medium power reactors in developing countries. These Guidelines were established during the Advisory Group Meeting held in Vienna from 11 to 15 May 1987. Their purpose is to review key aspects relating to the introduction of small and medium power reactors in developing countries; (2) up-dated information on SMPR Concepts Contributed by Supplier Industries. According to the recommendations of the Second Technical Committee Meeting on SMPRs held in Vienna in March 1985, this part contains the up-dated information formerly published in Annex 1 of the above mentioned TECDOC-347.

  5. Casimir force in the presence of a medium

    International Nuclear Information System (INIS)

    Kheirandish, Fardin; Soltani, Morteza; Sarabadani, Jalal

    2010-01-01

    We investigate the Casimir effect in the presence of a medium by quantizing the electromagnetic field in the presence of a magnetodielectric medium using the path-integral technique. For a given medium with definite electric and magnetic susceptibilities, explicit expressions for the Casimir force are obtained. The Lifshitz formula is recovered and in the absence of a medium the results tend to the original Casimir force between two conducting parallel plates immersed in the quantum electromagnetic vacuum.

  6. Degradation testing of Mg alloys in Dulbecco's modified eagle medium: Influence of medium sterilization

    International Nuclear Information System (INIS)

    Marco, Iñigo; Feyerabend, Frank; Willumeit-Römer, Regine; Van der Biest, Omer

    2016-01-01

    This work studies the in vitro degradation of Mg alloys for bioabsorbable implant applications under near physiological conditions. For this purpose, the degradation behaviour of Mg alloys in Dulbecco's modified eagle medium (DMEM) which is a commonly used cell culture medium is analysed. Unfortunately, DMEM can be contaminated by microorganisms, acidifying the medium and accelerating the Mg degradation process by dissolution of protective degradation layers, such as (Mg_x,Ca_y)(PO_4)_z. In this paper the influence of sterilization by applying UV-C radiation and antibiotics (penicillin/streptomycin) is analysed with two implant material candidates: Mg–Gd and Mg–Ag alloys; and pure magnesium as well as Mg–4Y–3RE as a reference. - Highlights: • Contamination of DMEM by microorganisms increases the degradation rate of Mg. • Mg and its alloys show passivation during long term immersion tests in DMEM. • The use of a control sample position is essential to assess H_2 evolution in DMEM.

  7. Incompatibility of Contrast Medium and Trisodium Citrate

    International Nuclear Information System (INIS)

    Delcour, Christian; Bruninx, Guy

    2013-01-01

    To test the compatibility of trisodium citrate, a catheter lock solution, with iodinated contrast medium. Iohexol, iobitridol, iodixanol, ioxaglate, ioxithalamate, iomeprol, and iopromide were tested. In all tests, 2 ml of contrast medium were mixed with 2 ml of trisodium citrate solution. Iodixanol and ioxaglate provoked a highly viscous gluelike precipitation when mixed with trisodium citrate. A brief transient precipitate was observed with iohexol, iomeprol, and ioxithalamate. Permanent precipitation occurred with iobitridol and iopromide. One must be aware of the potential for precipitation when contrast medium is mixed with trisodium citrate solution. Before trisodium citrate solution is injected, the catheter should be thoroughly flushed with saline if a contrast medium has previously been injected through it.

  8. IL-17 inhibits chondrogenic differentiation of human mesenchymal stem cells.

    Directory of Open Access Journals (Sweden)

    Masahiro Kondo

    Full Text Available OBJECTIVE: Mesenchymal stem cells (MSCs can differentiate into cells of mesenchymal lineages, such as osteoblasts and chondrocytes. Here we investigated the effects of IL-17, a key cytokine in chronic inflammation, on chondrogenic differentiation of human MSCs. METHODS: Human bone marrow MSCs were pellet cultured in chondrogenic induction medium containing TGF-β3. Chondrogenic differentiation was detected by cartilage matrix accumulation and chondrogenic marker gene expression. RESULTS: Over-expression of cartilage matrix and chondrogenic marker genes was noted in chondrogenic cultures, but was inhibited by IL-17 in a dose-dependent manner. Expression and phosphorylation of SOX9, the master transcription factor for chondrogenesis, were induced within 2 days and phosphorylated SOX9 was stably maintained until day 21. IL-17 did not alter total SOX9 expression, but significantly suppressed SOX9 phosphorylation in a dose-dependent manner. At day 7, IL-17 also suppressed the activity of cAMP-dependent protein kinase A (PKA, which is known to phosphorylate SOX9. H89, a selective PKA inhibitor, also suppressed SOX9 phosphorylation, expression of chondrogenic markers and cartilage matrix, and also decreased chondrogenesis. CONCLUSIONS: IL-17 inhibited chondrogenesis of human MSCs through the suppression of PKA activity and SOX9 phosphorylation. These results suggest that chondrogenic differentiation of MSCs can be inhibited by a mechanism triggered by IL-17 under chronic inflammation.

  9. Jet-induced medium excitation in heavy-ion collisions

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Wei [Key Laboratory of Quark and Lepton Physics (MOE) and Institute of Particle Physics, Central China Normal University, Wuhan 430079 (China); Pang, Long-Gang [Frankfurt Institute for Advanced Studies, Ruth-Moufang-Strasse 1, 60438 Frankfurt am Main (Germany); Stoecker, Horst [Frankfurt Institute for Advanced Studies, Ruth-Moufang-Strasse 1, 60438 Frankfurt am Main (Germany); Gesellschaft für Schwehrionenforschung, Planckstr. 1, Darmstadt (Germany); Luo, Tan; Wang, Enke [Key Laboratory of Quark and Lepton Physics (MOE) and Institute of Particle Physics, Central China Normal University, Wuhan 430079 (China); Wang, Xin-Nian [Key Laboratory of Quark and Lepton Physics (MOE) and Institute of Particle Physics, Central China Normal University, Wuhan 430079 (China); Nuclear Science Division Mailstop 70R0319, Lawrence Berkeley National Laboratory, Berkeley, CA 94740 (United States)

    2016-12-15

    We use a Linear Boltzmann Transport (LBT) model coupled to the (3+1)D ideal hydrodynamic evolution in real time with fluctuating initial conditions to simulate both the transport of jet shower partons and jet-induced medium excitation. In this coupled approach, propagation of energetic shower partons are treated in the LBT model with the 3+1D hydrodynamic model providing the evolving bulk medium. Soft partons from both elastic and inelastic processes in the LBT are fed back into the medium as a source term in the 3+1D hydrodynamics leading to induced medium excitation. We study the effect of jet-induced medium excitation via γ-hadron correlation within this coupled LBT-hydro (CoLBT-hydro) approach.

  10. van der Waals interactions in a magnetodielectric medium

    International Nuclear Information System (INIS)

    Spagnolo, S.; Dalvit, D. A. R.; Milonni, P. W.

    2007-01-01

    The van der Waals interaction between two ground-state atoms is calculated for two electrically or magnetically polarizable particles embedded in a dispersive magnetodielectric medium. Unlike previous calculations which infer the atom-atom interaction from the dilute-medium limit of the macroscopic, many-body van der Waals interaction, the interaction is calculated directly for the system of two atoms in a magnetodielectric medium. Two approaches are presented, the first based on the quantized electromagnetic field in a dispersive medium without absorption and the second on Green functions that allow for absorption. We show that the correct van der Waals interactions are obtained regardless of whether absorption in the host medium is explicitly taken into account

  11. AN INVESTIGATION INTO MEDIUM-SIZED MULTINATIONAL ENTERPRISES

    Directory of Open Access Journals (Sweden)

    Daniele Schilirò

    2013-09-01

    Full Text Available The paper provides an investigation of medium-sized Italian industrial enterprises that have become multinational companies. It concetrates on the set of medium and medium-large enterprises who seem to grow more in foreign markets, either through exports or through foreign direct investment. The work also offers a descriptive empirical picture of the performance of medium-sized Italian multinationals, which is compared with the performance of large corporations. From this analysis, which is based on several data sources, it is possible to outline a profile regarding the medium-size italian multinational enterprises; the aim is to understand the complex strategy towards internationalization of these companies, where the dimension of production is important and, therefore, innovation has a key role. Also the commercial dimension is crucial, because it leads to point to the direct supervision of foreign markets and to look very carefully at the customers, offering them a wide range of services. Finally, the paper highlights some critical issues that the medium sized multinational enterprises have to face for competing: namely, the stagnant productivity, the high taxation, the insufficient institutional support for internationalization, the bureaucracy and its high costs, the lack of skilled human capital available in the labor market due to inadequate policy training.

  12. A new selective chromogenic reagent for the spectrophotometric determination of thallium(I) and (III) and its separation using flotation and the solid-phase extraction on polyurethane foam.

    Science.gov (United States)

    Abou-El-Sherbini, Khaled S; Mostafa, Gamal A E; Hassanien, Mohamed M

    2003-09-01

    A new sensitive chromogenic reagent, 9,10-phenanthaquinone monoethylthiosemicarbazone (PET), has been synthesized and used in the spectrophotometric determination of Tl(III). In HNO3, H2SO4 or H3PO4 acids, PET can react immediately at room temperature with Tl(III) to form a red 2:1 complex with a maximum absorption at 516 nm. The different analytical parameters affecting the extraction and determination processes have been examined. The calibration curve was found to be linear over the range 0.2-10 microg cm(-3) with a molar absorptivity of 2.2 x 10(4) dm3 mol(-1) cm(-1). Sandell's sensitivity was found to be 0.0093 microg cm(-2). No interference from macroamounts of foreign ions was detected, except for Pd(II). However, Pd(II) does not affect the determination process, because its complex with PET has its lambda(max) at 625 nm. The proposed method has been applied to the determination of Tl(I and III) in synthetic and natural samples after separation by flotation (in oleic acid/kerosene) and solid-phase extraction (on polyurethane foam) techniques. The two methods were found to be accurate and not subject to random error, but solid-phase extraction was preferred because it is cheap, simpler and there is no contamination risk coming from flotation reagents.

  13. Diffusion with space memory modelled with distributed order space fractional differential equations

    Directory of Open Access Journals (Sweden)

    M. Caputo

    2003-06-01

    Full Text Available Distributed order fractional differential equations (Caputo, 1995, 2001; Bagley and Torvik, 2000a,b were fi rst used in the time domain; they are here considered in the space domain and introduced in the constitutive equation of diffusion. The solution of the classic problems are obtained, with closed form formulae. In general, the Green functions act as low pass fi lters in the frequency domain. The major difference with the case when a single space fractional derivative is present in the constitutive equations of diffusion (Caputo and Plastino, 2002 is that the solutions found here are potentially more fl exible to represent more complex media (Caputo, 2001a. The difference between the space memory medium and that with the time memory is that the former is more fl exible to represent local phenomena while the latter is more fl exible to represent variations in space. Concerning the boundary value problem, the difference with the solution of the classic diffusion medium, in the case when a constant boundary pressure is assigned and in the medium the pressure is initially nil, is that one also needs to assign the fi rst order space derivative at the boundary.

  14. Isolation, expansion and differentiation of cellular progenitors obtained from dental pulp of agouti (Dasyprocta prymnolopha Wagler, 1831

    Directory of Open Access Journals (Sweden)

    Yulla K.P. de Carvalho

    2015-06-01

    Full Text Available Abstract: The study aimed to isolate, expand, differentiate and characterize progenitor cells existent in the dental pulp of agouti. The material was washed with PBS solution and dissociated mechanically with the aid of a scalpel blade on plates containing culture medium D-MEM/F-12, and incubated at 5% CO2-37⁰C. The growth curve, CFU assay, osteogenic/adipogenic differentiation and characterization were obtained from the isolation. The cells began to be released from the explant tissue around the 7th day of culture. By day 22 of culture, cells reached 80% confluence. At the UFC test, 81 colonies were counted with 12 days of cultivation. The growth curves before and after freezing showed a regular growth with intense proliferation and clonogenic potential. The cell differentiation showed formation of osteoblasts and fat in culture, starting at 15 days of culture in a specific medium. Flow cytometry (FACs was as follows: CD34 (positive, CD14 (negative, CD45 (negative, CD73 (positive, CD79 (negative, CD90 (positive, CD105 (positive, demonstrating high specificity and commitment of isolated cells with mesenchymal stem cells strains. These results suggest the existence of a cell population of stem cells with mesenchymal features from the isolated tissue in the explants of agouti dental pulp, a potential model for study of stem cell strains obtained from the pulp tissue.

  15. Isolation of skin-derived precursors from human foreskin and their differentiation into neurons and glial cells

    Directory of Open Access Journals (Sweden)

    Bakhtiari M

    2010-12-01

    Full Text Available "n Normal 0 false false false EN-US X-NONE AR-SA MicrosoftInternetExplorer4 /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:Arial; mso-bidi-theme-font:minor-bidi;} Background: Skin-derived precursors (SKPs are a type of progenitor cells extracted from mammalian dermal tissue and can be differentiate to neural and mesodermal lineage in vitro. These cells can introduce an accessible autologos source of neural precursor cells for treatment of different neurodegenerative diseases. This research was done in order to set up isolation, culture, proliferation and differentiation of human skin derived precursors (hSKPs."n"nMethods: Human foreskin samples were cut into smaller pieces and cultured in proliferation medium after enzymatic digestion. To induce neural differentiation, cells were cultured in neural differentiation medium after fifth passage. We used immunocytochemistry and RT-PCR for characterization of the cells. Neuron and glial cell differentiation potential was assessed by immunofloresence using specific antibodies. The experiments were carried out in triplicate."n"nResults: After differentiation, βΙΙΙ- tubulin and neurofilament-M positive cells were observed that are specific markers for neurons. Moreover, glial fibrillary acid protein (GFAP and S100 positive cells were identified that are markers specifically express in glial cells. Detected neurons and glials were

  16. The effects of protoype medium on usability testing.

    Science.gov (United States)

    Boothe, Chase; Strawderman, Lesley; Hosea, Ethan

    2013-11-01

    Inconsistencies among testing methods and results in previous research prompted this study that builds upon a systematic usability testing research framework to better understand how interface medium influences users' abilities to detect usability flaws in applications. Interface medium was tested to identify its effects on users' perceptions of usability and abilities to detect usability problems and severe usability problems. Results indicated that medium has no effect on users' abilities to detect usability problems or perceptions of usability. However, results did indicate an interaction between the medium and the tested application in which users were able to identify significantly more usability problems on a higher fidelity medium using a particular application. Results also indicated that as users' perceptions of an application's usability increases, the users are less able to detect usability problems in that application. Usability testing should begin early in the design process, even if low fidelity mediums will be used. Copyright © 2013 Elsevier Ltd and The Ergonomics Society. All rights reserved.

  17. IMPLEMENTING SMALL AND MEDIUM IT PROJECTS IN SMALL AND MEDIUM ENERPRISES

    Directory of Open Access Journals (Sweden)

    Felix DUMITRESCU

    2014-06-01

    Full Text Available Information technology is essential nowadays for all companies. Small enterprises are an important part of the economy and this article aims at providing some useful insight in implementing modern IT projects to their benefit. Due to the limited funding available for the IT infrastructure in most start-ups and small businesses, the projects should be adapted to fulfill the needs of the company for the lowest cost. The paper will start by defining small and medium project management theory and outlining the target of the study, small and medium sized companies. Next it will show a number of case studies of IT projects implemented in different types of small companies in Romania. Based on these implementations the article will draw some conclusions relevant to most small companies which need to design or improve their IT infrastructure.

  18. Medium effects in direct reactions

    International Nuclear Information System (INIS)

    Karakoc, M; Bertulani, C

    2013-01-01

    We discuss medium corrections of the nucleon-nucleon (NN) cross sections and their influence on direct reactions at intermediate energies ≳50 MeV/nucleon. The results obtained with free NN cross sections are compared with those obtained with a geometrical treatment of Pauli-blocking and Dirac-Bruecker methods. We show that medium corrections may lead to sizable modifications for collisions at intermediate energies and that they are more pronounced in reactions involving weakly bound nuclei.

  19. Cultivation of tea fungus on malt extract medium

    Directory of Open Access Journals (Sweden)

    Cvetković Dragoljub D.

    2002-01-01

    Full Text Available The possibility of application of malt extract as a source of carbohydrate in a medium for tea fungus was investigated. The beverage obtained on such medium was compared with that prepared in a traditional way with sucrose medium. The presence of easily adoptable sugars, glucose and fructose, as dominant in malt medium results in a very effective fermentation, which gives much more sour beverage for the same time and makes it possible to reduce the fermentation period. The obtained beverage has satisfactory sensorial characteristics.

  20. System and method for treatment of a medium

    Science.gov (United States)

    Singh, Surinder Prabhjot; Acharya, Harish Radhakrishna; Perry, Robert James; McDermott, John Brian

    2017-05-23

    A system and method for treatment of a medium is disclosed. The system includes a plurality of separator zones and a plurality of heat transfer zones. Each of the separator zone and the heat transfer zone among the plurality of separator zones and heat transfer zones respectively, are disposed alternatively in a flow duct. Further, each separator zone includes an injector device for injecting a sorbent into the corresponding separator zone. Within the corresponding separator zone, the injected sorbent is reacted with a gaseous medium flowing in the flow duct, so as to generate a reacted gaseous medium and a reacted sorbent. Further, each heat transfer zone exchanges heat between the reacted gaseous medium fed from the corresponding separator zone and a heat transfer medium.

  1. Evaluating Pharmacy Student Attitudes Toward the Medium of Comics for Providing Information on Adult Immunizations

    Directory of Open Access Journals (Sweden)

    Jagannath M. Muzumdar

    2015-01-01

    Full Text Available Objectives: This study aimed at designing and assessing educational materials for adult immunizations through the medium of comics. The study design evaluated the effects of two vaccine information flyers (a CDC flyer vs a flyer designed in Comic medium on participants’ attitude towards the flyers. Methods: A between-group, randomized trial was used to compare the effectiveness of two vaccine information flyers on participants’ attitude towards the flyers. Upon approval from the human subjects review committee, student participants (age ≥18 years were randomly assigned either the CDC or comic flyer. They were then asked to respond to survey items developed to measure the flyers’ effect on participants’ attitude towards the flyer. Items were measured using a 7-point semantic differential scale. Cronbach’s alpha was computed for reliability testing of the study instrument. Independent-samples t-test was used to compare means of the two groups with respect to their attitudes toward the flyer. Results: A total of 170 third-year pharmacy students participated in the study (N = 91 for Comic flyer & N = 79 for CDC flyer. There was a significant difference in attitudes toward the flyer between students who read comic flyers (mean = 6.14; SD = 0.62 and those who read CDC flyers (mean = 4.93; SD = 1.20. Additional comments provided by students further confirmed the quantitative findings of the study. Student responses to the use of comics as a medium of providing information on adult immunization were constructive and encouraging. Conclusion: The study findings showed that the flyer with comics was evaluated more attractive than the CDC flyer. The positive findings of our study could provide a new direction for developing educational materials about adult immunizations. Future research on comics, as a medium of communication, could explore its use as a tool for providing healthcare information to consumers.   Type: Original Research

  2. Analysis of the behavior of radionuclides migration in fractured medium in different types of rocks matrices

    International Nuclear Information System (INIS)

    Sá, Ludimila Silva Salles de; Silveira, Cláudia Siqueira da; Lima, Zelmo Rodrigues de

    2017-01-01

    In management of radioactive wastes, the current trend is to dispose the radioactive waste for long life and high activity in permanent repositories of depth, geologically stable and low permeability. Thus, it is relevant to analyze the groundwater movement process, because the mechanism by which the radionuclides in a repository with fractures could return to the surface would be through the groundwater circulation system. A common problem encountered is the modeling of the migration of radionuclides in a fractured medium. The objective of this work is to evaluate the behavior of the migration of radionuclides in two types of rock matrix, considering the following properties: volumetric density, porosity, distribution coefficient and molecular diffusion coefficient. The physical system adopted consists of the matrix rock containing a discrete fracture in a porous medium saturated with water. The partial differential equations that describe the radionuclide movement were discretized by finite differences, and the Implicit Euler method was adopted. While for the convective term the numerical scheme of progressive differences was used

  3. Quantitative real-time RT-PCR and chromogenic in situ hybridization: precise methods to detect HER-2 status in breast carcinoma

    Directory of Open Access Journals (Sweden)

    Soares Fernando A

    2009-03-01

    Full Text Available Abstract Background HER-2 gene testing has become an integral part of breast cancer patient diagnosis. The most commonly used assay in the clinical setting for evaluating HER-2 status is immunohistochemistry (IHC and fluorescence in situ hybridization (FISH. These procedures permit correlation between HER-2 expression and morphological features. However, FISH signals are labile and fade over time, making post-revision of the tumor difficult. CISH (chromogenic in situ hybridization is an alternative procedure, with certain advantages, although still limited as a diagnostic tool in breast carcinomas. Methods To elucidate the molecular profile of HER-2 status, mRNA and protein expression in 75 invasive breast carcinomas were analyzed by real time quantitative RT-PCR (qRT-PCR and IHC, respectively. Amplifications were evaluated in 43 of these cases by CISH and in 11 by FISH. Results The concordance rate between IHC and qRT-PCR results was 78.9%, and 94.6% for qRT-PCR and CISH. Intratumoral heterogeneity of HER-2 status was identified in three cases by CISH. The results of the three procedures were compared and showed a concordance rate of 83.8%; higher discordances were observed in 0 or 1+ immunostaining cases, which showed high-level amplification (15.4% and HER-2 transcript overexpression (20%. Moreover, 2+ immunostaining cases presented nonamplified status (50% by CISH and HER-2 downexpression (38.5% by qRT-PCR. In general, concordance occurred between qRT-PCR and CISH results. A high concordance was observed between CISH/qRT-PCR and FISH. Comparisons with clinicopathological data revealed a significant association between HER-2 downexpression and the involvement of less than four lymph nodes (P = 0.0350. Conclusion Based on these findings, qRT-PCR was more precise and reproducible than IHC. Furthermore, CISH was revealed as an alternative and useful procedure for investigating amplifications involving the HER-2 gene.

  4. Development of global medium-energy nucleon-nucleus optical model potentials

    International Nuclear Information System (INIS)

    Madland, D.G.; Sierk, A.J.

    1997-01-01

    The authors report on the development of new global optical model potentials for nucleon-nucleus scattering at medium energies. Using both Schroedinger and Dirac scattering formalisms, the goal is to construct a physically realistic optical potential describing nucleon-nucleus elastic scattering observables for a projectile energy range of (perhaps) 20 meV to (perhaps) 2 GeV and a target mass range of 16 to 209, excluding regions of strong nuclear deformation. They use a phenomenological approach guided by conclusions from recent microscopic studies. The experimental database consists largely of proton-nucleus elastic differential cross sections, analyzing powers, spin-rotation functions, and total reaction cross sections, and neutron-nucleus total cross sections. They will use this database in a nonlinear least-squares adjustment of optical model parameters in both relativistic equivalent Schroedinger (including relativistic kinematics) and Dirac (second-order reduction) formalisms. Isospin will be introduced through the standard Lane model and a relativistic generalization of that model

  5. Selective medium for culture of Mycoplasma hyopneumoniae.

    Science.gov (United States)

    Cook, Beth S; Beddow, Jessica G; Manso-Silván, Lucía; Maglennon, Gareth A; Rycroft, Andrew N

    2016-11-15

    The fastidious porcine respiratory pathogen Mycoplasma hyopneumoniae has proven difficult to culture since it was first isolated in 1965. A reliable solid medium has been particularly challenging. Moreover, clinical and pathological samples often contain the fast-growing M. hyorhinis which contaminates and overgrows M. hyopneumoniae in primary culture. The aim of this study was to optimise the culture medium for recovery of M. hyopneumoniae and to devise a medium for selection of M. hyopneumoniae from clinical samples also containing M. hyorhinis. The solid medium devised by Niels Friis was improved by use of Purified agar and incorporation of DEAE-dextran. Addition of glucose or neutralization of acidity in liquid medium with NaOH did not improve the final yield of viable organisms or alter the timing of peak viability. Analysis of the relative susceptibility of M. hyopneumoniae and M. hyorhinis strains to four antimicrobials showed that M. hyopneumoniae is less susceptible than M. hyorhinis to kanamycin. This was consistent in all UK and Danish strains tested. A concentration of 2μg/ml of kanamycin selectively inhibited the growth of all M. hyorhinis tested, while M. hyopneumoniae was able to grow. This forms the basis of an effective selective culture medium for M. hyopneumoniae. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

  6. Map of fluid flow in fractal porous medium into fractal continuum flow.

    Science.gov (United States)

    Balankin, Alexander S; Elizarraraz, Benjamin Espinoza

    2012-05-01

    This paper is devoted to fractal continuum hydrodynamics and its application to model fluid flows in fractally permeable reservoirs. Hydrodynamics of fractal continuum flow is developed on the basis of a self-consistent model of fractal continuum employing vector local fractional differential operators allied with the Hausdorff derivative. The generalized forms of Green-Gauss and Kelvin-Stokes theorems for fractional calculus are proved. The Hausdorff material derivative is defined and the form of Reynolds transport theorem for fractal continuum flow is obtained. The fundamental conservation laws for a fractal continuum flow are established. The Stokes law and the analog of Darcy's law for fractal continuum flow are suggested. The pressure-transient equation accounting the fractal metric of fractal continuum flow is derived. The generalization of the pressure-transient equation accounting the fractal topology of fractal continuum flow is proposed. The mapping of fluid flow in a fractally permeable medium into a fractal continuum flow is discussed. It is stated that the spectral dimension of the fractal continuum flow d(s) is equal to its mass fractal dimension D, even when the spectral dimension of the fractally porous or fissured medium is less than D. A comparison of the fractal continuum flow approach with other models of fluid flow in fractally permeable media and the experimental field data for reservoir tests are provided.

  7. How Does the Medium Affect the Message?

    Science.gov (United States)

    Dommermuth, William P.

    1974-01-01

    This experimental comparison of the advertising effectiveness of television, movies, radio, and print finds no support for McLuhan's idea that television is a "cool" medium and movies are a "hot" medium. (RB)

  8. Medium modifications of vector mesons

    International Nuclear Information System (INIS)

    Pant, L.M.

    2004-01-01

    The omega photoproduction in nuclear medium with the ELSA facility at Bonn and the present status of the HADES collaboration to investigate the in-medium hadron properties in proton, heavy ions and hadron induced reactions at GSI, Darmstadt are presented. Efforts are under way to utilise the electron beam at Indore for experimental hadron physics in order to step into the intermediate energy nuclear physics regime. The skeletal outline of the high energy electron beam now available at CAT, Indore is discussed

  9. Hadrons in hot and dense medium

    International Nuclear Information System (INIS)

    Mallik, S.

    2004-01-01

    We review chiral perturbation theory in some detail and construct interaction terms involving the Goldstone and the different non-Goldstone fields, in presence of external (classical) fields coupled to currents. The ensemble average of the two-point functions of the currents can now be expanded in terms of Feynman diagrams. We evaluate the one-loop diagrams in the neighbourhood of the respective poles to find the effective couplings and masses of the particles in medium. We also describe the virial formula for the self-energy of a particle in medium, giving its pole position. It proves useful if the scattering amplitude of the particle with particles in medium is known experimentally. (author)

  10. An In Vitro Study of Differentiation of Hematopoietic Cells to Endothelial Cells

    Directory of Open Access Journals (Sweden)

    Qi Ru Wang

    2011-01-01

    medium (ECCM. BM-EPCs were characterized in terms of phenotype, lineage potential, and their functional properties. Endothelial cell colonies derived from BM-EPC were cultured with ECCM for 3 months. Cultured EPC colony cells expressed endothelial cell markers and formed the capillary-like network in vitro. EPC colony cells expressed differential proliferative capacity; some of the colonies exhibited a high proliferative potential (HPP capacity up to 20 population doublings. More importantly, these HPP-EPCs expressed hematopoietic marker CD45, exhibited endocytic activities, and preserved some of the myeloid cell activity. In addition, the HPP-EPCs secrete various growth factors including VEGF and GM-CSF into the culture medium. The results demonstrate that these EPCs were primarily derived from hematopoietic origin of early precursor cells and maintained high proliferative potential capacity, a feature with a significant potential in the application of cell therapy in ischemic diseases.

  11. [Effects of triterpenoid from Psidium guajava leaves ursolic acid on proliferation, differentiation of 3T3-L1 preadipocyte and insulin resistance].

    Science.gov (United States)

    Lin, Juan-Na; Kuang, Qiao-Ting; Ye, Kai-He; Ye, Chun-Ling; Huang, Yi; Zhang, Xiao-Qi; Ye, Wen-Cai

    2013-08-01

    To investigate the influences of triterpenoid from Psidium guajava Leaves (ursolic acid) on the proliferation, differentiation of 3T3-L1 preadipocyte, and its possible mechanism treat for insulin resistance. 3T3-L1 preadipocyte was cultured in vitro. After adding ursolic acid to the culture medium for 48h, the cell viability was tested by MTT assay. Induced for 6 days, the lipid accumulation of adipocyte was measured by Oil Red O staining. The insulin resistant cell model was established with Dexamethasone. Cellular glucose uptake was determined with GOD-POD assays and FFA concentration was determined at the time of 48h. Secreted adiponectin were measured by ELISA. The protein levels of PPARgamma and PTP1B in insulin resistant adipocyte were measured by Western Blotting. Compared with medium control group, 30, 100 micromol/L ursolic acid could increase its proliferation and differentiation significantly (P 0.05). Ursolic acid can improve the proliferation and differentiation of 3T3-L1 preadipocyte, enhance cellular glucose uptake, inhibit the production of FFA, promote the secretion of adiponectin insulin resistant adipocyte, its mechanism may be related to upregulating the expression of PPARgamma protein.

  12. The relative contribution of paracine effect versus direct differentiation on adipose-derived stem cell transplantation mediated cardiac repair.

    Directory of Open Access Journals (Sweden)

    Dezhong Yang

    Full Text Available BACKGROUND: Recent studies have demonstrated that transplantation of adipose-derived stem cell (ADSC can improve cardiac function in animal models of myocardial infarction (MI. However, the mechanisms underlying the beneficial effect are not fully understood. In this study, we characterized the paracrine effect of transplanted ADSC and investigated its relative importance versus direct differentiation in ADSC transplantation mediated cardiac repair. METHODOLOGY/PRINCIPAL FINDINGS: MI was experimentally induced in mice by ligation of the left anterior descending coronary artery. Either human ADSC, conditioned medium (CM collected from the same amount of ADSC or control medium was injected into the peri-infarct region immediately after MI. Compared with the control group, both ADSC and ADSC-CM significantly reduced myocardial infarct size and improved cardiac function. The therapeutic efficacy of ADSC was moderately superior to ADSC-CM. ADSC-CM significantly reduced cardiomyocyte apoptosis in the infarct border zone, to a similar degree with ADSC treatment. ADSC enhanced angiogenesis in the infarct border zone, but to a stronger degree than that seen in the ADSC-CM treatment. ADSC was able to differentiate to endothelial cell and smooth muscle cell in post-MI heart; these ADSC-derived vascular cells amount to about 9% of the enhanced angiogenesis. No cardiomyocyte differentiated from ADSC was found. CONCLUSIONS: ADSC-CM is sufficient to improve cardiac function of infarcted hearts. The therapeutic function of ADSC transplantation is mainly induced by paracrine-mediated cardioprotection and angiogenesis, while ADSC differentiation contributes a minor benefit by being involved in angiogenesis. Highlights 1 ADSC-CM is sufficient to exert a therapeutic potential. 2. ADSC was able to differentiate to vascular cells but not cardiomyocyte. 3. ADSC derived vascular cells amount to about 9% of the enhanced angiogenesis. 4. Paracrine effect is the major

  13. Generalized Lagrangian Jacobi Gauss collocation method for solving unsteady isothermal gas through a micro-nano porous medium

    Science.gov (United States)

    Parand, Kourosh; Latifi, Sobhan; Delkhosh, Mehdi; Moayeri, Mohammad M.

    2018-01-01

    In the present paper, a new method based on the Generalized Lagrangian Jacobi Gauss (GLJG) collocation method is proposed. The nonlinear Kidder equation, which explains unsteady isothermal gas through a micro-nano porous medium, is a second-order two-point boundary value ordinary differential equation on the unbounded interval [0, ∞). Firstly, using the quasilinearization method, the equation is converted to a sequence of linear ordinary differential equations. Then, by using the GLJG collocation method, the problem is reduced to solving a system of algebraic equations. It must be mentioned that this equation is solved without domain truncation and variable changing. A comparison with some numerical solutions made and the obtained results indicate that the presented solution is highly accurate. The important value of the initial slope, y'(0), is obtained as -1.191790649719421734122828603800159364 for η = 0.5. Comparing to the best result obtained so far, it is accurate up to 36 decimal places.

  14. Roentgen contrast medium

    International Nuclear Information System (INIS)

    Tamborski, C.

    1989-01-01

    The patent deals with a roentgen contrast medium containing a perfluorinebrominealkylether of the formula C m F 2m+1 OC n F 2n Br dispersed in water, preferentially in the presence of a non-ionic dispersing agent such as a fluorinated amidoaminoxide. 2 tabs

  15. Regulation of proliferation and differentiation of adipocyte precursor cells in rainbow trout (Oncorhynchus mykiss).

    Science.gov (United States)

    Bouraoui, L; Gutiérrez, J; Navarro, I

    2008-09-01

    Here, we describe optimal conditions for the culture of rainbow trout (Oncorhynchus mykiss) pre-adipocytes obtained from adipose tissue and their differentiation into mature adipocytes, in order to study the endocrine control of adipogenesis. Pre-adipocytes were isolated by collagenase digestion and cultured on laminin or 1% gelatin substrate. The expression of proliferating cell nuclear antigen was used as a marker of cell proliferation on various days of culture. Insulin growth factor-I stimulated cell proliferation especially on days 5 and 7 of culture. Tumor necrosis factor alpha (TNFalpha) slightly enhanced cell proliferation only at a low dose. We verified the differentiation of cells grown in specific medium into mature adipocytes by oil red O (ORO) staining. Quantification of ORO showed an increase in triglycerides throughout culture. Immunofluorescence staining of cells at day 11 revealed the expression of CCAAT/enhancer-binding protein and peroxisome proliferator-activator receptor gamma, suggesting that these transcriptional factors are involved in adipocyte differentiation in trout. We also examined the effect of TNFalpha on the differentiation of these adipocytes in primary culture. TNFalpha inhibited the differentiation of these cells, as indicated by a decrease in glycerol-3-phosphate dehydrogenase activity, an established marker of adipocyte differentiation. In conclusion, the culture system described here for trout pre-adipocytes is a powerful tool to study the endocrine regulation of adipogenesis in this species.

  16. Proteoglycan biosynthesis in murine monocytic leukemic (M1) cells before and after differentiation

    International Nuclear Information System (INIS)

    McQuillan, D.J.; Yanagishita, M.; Hascall, V.C.; Bickel, M.

    1989-01-01

    Murine monocytic leukemic (M1) cells were cultured in the presence of [ 3 H]glucosamine and [ 35 S]sulfate. Labeled proteoglycans were purified by anion exchange chromatography and characterized by gel filtration and sodium dodecyl sulfate-polyacrylamide gel electrophoresis in combination with chemical and enzymatic degradation. M1 cells synthesize a single predominant species of proteoglycan which distributes almost equally between the cell and medium after 17 h labeling. The cell-associated proteoglycan has an overall size of about 135 kDa and contains three to five chondroitin sulfate chains (28-31 kDa each) attached to a chondroitinase-generated core protein of 28 kDa. The synthesis and subsequent secretion of this proteoglycan was enhanced 4-5-fold in cells induced to differentiate into macrophages. This was not a phenomenon of arrest in the G0/G1 stage of the cell cycle, since density inhibited undifferentiated cells arrested at this stage did not increase proteoglycan synthesis. The chondroitin sulfate chains contained exclusively chondroitin 4- and 6-sulfate; however, the ratio of these two disaccharides differed between the medium- and cell-associated proteoglycans, and changed during progression of the cells into a fully differentiated phenotype. Pulse-chase kinetics indicate the presence of two distinct pools of proteoglycan; one that is secreted very rapidly from the cell after a approximately 1-h lag, and a second pool that is turned over in the cell with a half-time of approximately 3.5 h. Subtle differences in the glycosylation patterns of the medium- and cell-associated species are consistent with synthesis of two pools. Papain digestion suggests that the chondroitin sulfate chains are clustered on a small protease resistant peptide. The data suggest that this proteoglycan is similar to the serglycin proteoglycan family

  17. Medium-Term Health of Seniors Following Exposure to a Natural Disaster.

    Science.gov (United States)

    Labra, Oscar; Maltais, Danielle; Gingras-Lacroix, Gabriel

    2018-01-01

    The article aims to describe the medium-term impacts of a major earthquake event (Chile, February 27, 2010) on 26 seniors. The authors adopted a qualitative study approach. Data obtained using the Impact of Event Scale-Revised (IES-R) show the presence of manifestations of posttraumatic stress in the majority of respondents. In addition, data collected in interviews demonstrated a progressive deterioration of the health of respondents over a period of 4 years following the disaster. Seniors are particularly vulnerable to the effects of material loss, emotional stress, and postdisaster health complications. These impacts are exacerbated by low economic status. Furthermore, broader research is necessary involving elderly living in poverty who have survived natural disasters and others without such experiences, in order to better identify and differentiate between health complications associated with exposure to disaster events and those linked more strictly with natural aging processes.

  18. Overview of medium heterogeneity and transport processes

    International Nuclear Information System (INIS)

    Tsang, Y.; Tsang, C.F.

    1993-11-01

    Medium heterogeneity can have significant impact on the behavior of solute transport. Tracer breakthrough curves from transport in a heterogeneous medium are distinctly different from that in a homogeneous porous medium. Usually the shape of the breakthrough curves are highly non-symmetrical with a fast rise at early times and very long tail at late times, and often, they consist of multiple peaks. Moreover, unlike transport in a homogeneous medium where the same transport parameters describe the entire medium, transport through heterogeneous media gives rise to breakthrough curves which have strong spatial dependence. These inherent characteristics of transport in heterogeneous medium present special challenge to the performance assessment of a potential high level nuclear waste repository with respect to the possible release of radio nuclides to the accessible environment. Since an inherently desirable site characteristic for a waste repository is that flow and transport should be slow, then transport measurements in site characterization efforts will necessarily be spatially small and temporally short compare to the scales which are of relevance to performance assessment predictions. In this paper we discuss the role of medium heterogeneity in site characterization and performance assessment. Our discussion will be based on a specific example of a 3D heterogeneous stochastic model of a site generally similar to, the Aespoe Island, the site of the Hard Rock Laboratory in Southern Sweden. For our study, alternative 3D stochastic fields of hydraulic conductivities conditioned on ''point'' measurements shall be generated. Results of stochastic flow and transport simulations would be used to address the issues of (1) the relationship of tracer breakthrough with the structure of heterogeneity, and (2) the inference from small scale testing results to large scale and long term predictions

  19. CHROMagar Orientation Medium Reduces Urine Culture Workload

    Science.gov (United States)

    Manickam, Kanchana; Karlowsky, James A.; Adam, Heather; Lagacé-Wiens, Philippe R. S.; Rendina, Assunta; Pang, Paulette; Murray, Brenda-Lee

    2013-01-01

    Microbiology laboratories continually strive to streamline and improve their urine culture algorithms because of the high volumes of urine specimens they receive and the modest numbers of those specimens that are ultimately considered clinically significant. In the current study, we quantitatively measured the impact of the introduction of CHROMagar Orientation (CO) medium into routine use in two hospital laboratories and compared it to conventional culture on blood and MacConkey agars. Based on data extracted from our Laboratory Information System from 2006 to 2011, the use of CO medium resulted in a 28% reduction in workload for additional procedures such as Gram stains, subcultures, identification panels, agglutination tests, and biochemical tests. The average number of workload units (one workload unit equals 1 min of hands-on labor) per urine specimen was significantly reduced (P < 0.0001; 95% confidence interval [CI], 0.5326 to 1.047) from 2.67 in 2006 (preimplementation of CO medium) to 1.88 in 2011 (postimplementation of CO medium). We conclude that the use of CO medium streamlined the urine culture process and increased bench throughput by reducing both workload and turnaround time in our laboratories. PMID:23363839

  20. Human periapical cyst-mesenchymal stem cells differentiate into neuronal cells.

    Science.gov (United States)

    Marrelli, M; Paduano, F; Tatullo, M

    2015-06-01

    It was recently reported that human periapical cysts (hPCys), a commonly occurring odontogenic cystic lesion of inflammatory origin, contain mesenchymal stem cells (MSCs) with the capacity for self-renewal and multilineage differentiation. In this study, periapical inflammatory cysts were compared with dental pulp to determine whether this tissue may be an alternative accessible tissue source of MSCs that retain the potential for neurogenic differentiation. Flow cytometry and immunofluorescence analysis indicated that hPCy-MSCs and dental pulp stem cells spontaneously expressed the neuron-specific protein β-III tubulin and the neural stem-/astrocyte-specific protein glial fibrillary acidic protein (GFAP) in their basal state before differentiation occurs. Furthermore, undifferentiated hPCy-MSCs showed a higher expression of transcripts for neuronal markers (β-III tubulin, NF-M, MAP2) and neural-related transcription factors (MSX-1, Foxa2, En-1) as compared with dental pulp stem cells. After exposure to neurogenic differentiation conditions (neural media containing epidermal growth factor [EGF], basic fibroblast growth factor [bFGF], and retinoic acid), the hPCy-MSCs showed enhanced expression of β-III tubulin and GFAP proteins, as well as increased expression of neurofilaments medium, neurofilaments heavy, and neuron-specific enolase at the transcript level. In addition, neurally differentiated hPCy-MSCs showed upregulated expression of the neural transcription factors Pitx3, Foxa2, Nurr1, and the dopamine-related genes tyrosine hydroxylase and dopamine transporter. The present study demonstrated for the first time that hPCy-MSCs have a predisposition toward the neural phenotype that is increased when exposed to neural differentiation cues, based on upregulation of a comprehensive set of proteins and genes that define neuronal cells. In conclusion, these results provide evidence that hPCy-MSCs might be another optimal source of neural/glial cells for cell