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Sample records for surface tissue inoculated

  1. Microbiological and sensory effects of the combined application of hot-cold organic acid sprays and steam condensation at subatmospheric pressure for decontamination of inoculated pig tissue surfaces.

    Science.gov (United States)

    Smulders, Frans J M; Wellm, Gabriele; Hiesberger, Johann; Rohrbacher, Irene; Bauer, Alexandra; Paulsen, Peter

    2011-08-01

    We studied microbiological and sensory effects of treating pig tissue for 15 s with 55 and 10°C sprays of acetic acid (AA; 0.15 to 0.3 M) and lactic acid (LA; 0.1 to 0.2 M) solutions prior to the tissue being subjected to steam condensation (18 s at 65°C or 10 s at 75°C). LA or AA spraying and then steam treatment resulted in 3- to 4-log average reductions of Pseudomonas fragi and Yersinia enterocolitica inocula (6 to 7 log CFU/cm(2)), regardless of acid temperature or concentration. Buffered LA or 1:1 mixtures of AA:LA and then steam treatment yielded similar reductions. Most of the acid-steam-treated samples had microbial counts below the limit of detection (2 log CFU/cm(2)); thus, the results likely underestimate the potential of this procedure. When the period between inoculation and acid-steam treatment was extended from 0.5 to 24 h, up to a 1-log-higher microbial reduction was observed, due to a 1- to 2-log-greater initial contamination. Increasing the LA contact time to 6 min increased the microbial reduction by 0.8 log. Acid-steam treatment effected lower L* values (darker color) on pigskin, but higher L* values on muscle and fat tissue (paler color). Many muscle samples exhibited lower a* values and off-color brown hues. Off-odors were observed immediately after treatment, but with the exception of fat tissue and AA-treated samples, they largely disappeared during further storage. Off-flavors were only detected in AA-treated muscle samples.

  2. Automatic surface inoculation of agar trays.

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    Wilkins, J. R.; Mills, S. M.; Boykin, E. H.

    1972-01-01

    Description of a machine and technique for the automatic inoculation of a plastic tray containing agar media with a culture, using either a conventional inoculation loop or a cotton swab. The design of the machine is simple, it is easy to use, and it relieves the operator from the manual task of streaking cultures. The described technique makes possible the visualization of the overall qualitative and, to some extent, quantitative relationships of various bacterial types in a sample tested.

  3. IN VITRO INOCULATION OF ASPARAGUS OFFICINALIS TISSUE CULTURE SHOOTS WITH FUSARIUM PROLIFERA TUM

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    A.K.MoHD OMAR

    1999-01-01

    Full Text Available Artificially inoculated asparagus tissue culture plantlets with a virulent fungus, Fusarium proliferatum showed signs of infection as early as 4 days after inoculat ion. Macroscopic observations revealed presence of early symptoms such as necrotic lesions at the affected area and light microscopic examinations clearly revealed the post-penetration events that took place including the destruction of surrounding cells. However, little is known of the hyphal activity or advancement on the host's surface at the initial stage after inoculation. Scanning electron microscopic examination clearly revealed the hyphal advancement on the surface and the mode of entrance into the host tissues beneath. Four days after inoculation, the fungi proceeded to spread out from the inoculation point onto the host surface which eventually developed into a sparse network of both aerial and non-aerial hyphae. Non-aerial hyphae form a network of mycelium that adheres to the surface and it's movement appeared to be oriented towards the stomata. Hyphal penetration occurs more often through the stomata, natural openings or wounds. In some cases, the hyphae crossed over the stomatal opening w ithout entering the host tissues. At places where the cuticle layer is absent or not well developed the hyphae successfully grew in between the epidermal cells into the tissues beneath.

  4. Tissue reactions to bacteria-inoculated rat lead samples .2. Effect of local gentamicin release through surface-modified polyurethane tubing

    NARCIS (Netherlands)

    vanWachem, PB; vanLuyn, MJA; deWit, AW; Raatjes, D; Hendriks, M; Verhoeven, MLPM; Cahalan, PT

    1997-01-01

    A surface modification technique was developed to achieve controlled release of gentamicin from implanted polyurethane (PU) rat lead samples. PU tubing first was provided with an acrylic acid/acrylamide copolymer surface graft and then loaded with gentamicin. This surface modification technique resu

  5. Antimicrobial Efficacy of a Lactic Acid and Citric Acid Blend against Shiga Toxin-Producing Escherichia coli, Salmonella, and Nonpathogenic Escherichia coli Biotype I on Inoculated Prerigor Beef Carcass Surface Tissue.

    Science.gov (United States)

    Scott, Brittney R; Yang, Xiang; Geornaras, Ifigenia; Delmore, Robert J; Woerner, Dale R; Adler, Jeremy M; Belk, Keith E

    2015-12-01

    Studies were conducted to (i) determine whether inoculants of nonpathogenic Escherichia coli biotype I effectively served as surrogates for E. coli O157:H7, non-O157 Shiga toxin-producing E. coli, and Salmonella when prerigor beef carcass tissue was treated with a commercially available blend of lactic acid and citric acid (LCA) at a range of industry conditions of concentration, temperature, and pressure; (ii) determine the antimicrobial efficacy of LCA; and (iii) investigate the use of surrogates to validate a hot water and LCA sequential treatment as a carcass spray intervention in a commercial beef harvest plant. In an initial laboratory study, beef brisket tissue samples were left uninoculated or were inoculated (∼6 log CFU/cm(2)) on the adipose side with E. coli O157:H7 (5-strain mixture), non-O157 Shiga toxin-producing E. coli (12-strain mixture), Salmonella (6-strain mixture), or nonpathogenic E. coli (5-strain mixture). Samples were left untreated (control) or were treated with LCA, in a spray cabinet, at one of eight combinations of solution concentration (1.9 and 2.5%), solution temperature (43 and 60°C), and application pressure (15 and 30 lb/in(2)). In a second study, the E. coli surrogates were inoculated (∼6 log CFU/cm(2)) on beef carcasses in a commercial facility to validate the use of a hot water treatment (92.2 to 92.8°C, 13 to 15 lb/in(2)) followed by an LCA treatment (1.9%, 50 to 51.7°C, 13 to 15 lb/in(2), 10 s). In the in vitro study, surrogate and pathogen bacteria did not differ in their response to the tested LCA treatments. Treatment with LCA reduced (P < 0.05) inoculated populations by 0.9 to 1.5 log CFU/cm(2), irrespective of inoculum type. The hot water and LCA sequential treatments evaluated in the commercial facility reduced (P < 0.05) the inoculated nonpathogenic E. coli surrogates on carcasses by 3.7 log CFU/cm(2). This study therefore provides the meat industry with data for this sequential multiple hurdle system for the

  6. Toxoplasma gondii infections in red-tailed hawks inoculated orally with tissue cysts.

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    Lindsay, D S; Dubey, J P; Blagburn, B L

    1991-04-01

    The response to inoculation of Toxoplasma gondii tissue cysts was examined in 3 red-tailed hawks (Buteo jamaicensis). One hawk (hawk 1) was inoculated orally with 3.000 tissue cysts of the GT-1 isolate of T. gondii and 2 hawks (hawks 2 and 3) each were inoculated orally with 12,000 tissue cysts of a mixture of 8 isolates of T. gondii. None of the hawks developed clinical signs of toxoplasmosis. Serum antibodies were measured with the modified direct agglutination test using formalin-fixed tachyzoites. Hawk 1 had a titer of 1:40 prior to inoculation and did not have an increase in titer during the study. Hawks 2 and 3 had titers of 1:5 and 1:10, respectively, prior to inoculation, and both had increased titers (titers greater than or equal to 1:60) by 1 wk postinoculation and remained T. gondii antibody positive throughout the 10 wk of the study. Toxoplasma gondii was isolated from the heart and breast muscle of hawk 1. The biologic behavior of this T. gondii isolate was different from the 1 inoculated, and it probably represents a prior natural infection. Toxoplasma gondii was isolated from the brain, heart, breast muscle, and a mixture of gizzard and proventriculus from hawk 2 and from breast muscle of hawk 3. Toxoplasma gondii was not isolated from the eye, lung, liver, kidney, or spleen of any red-tailed hawk.

  7. [Rabies Tissue Culture Infection Test as an Alternative for the Mouse Inoculation Test

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    Zanoni, R.; Hörnlimann, B.; Wandeler, A. I.; Kappeler, A.; Kipfer, R.; Peterhans, E.

    1990-01-01

    Rabies has disappeared from large parts of Switzerland. Due to systematic oral fox-vaccination campaings that started in 1987, cases of rabies in wild and domestic animals have been confined to the western frontier with France in the last three years. Nevertheless, some cases of severe exposition of man by rabid or rabies-suspect animals still occur. Rabies can be diagnosed in brain smears of infected animals with high specificity and sensitivity by a direct immunofluorescence method. According to WHO recommendations, negative results are to be confirmed in cases of a human exposition by intracerebral inoculation of brain suspensions in three-weeks-old mice. This method has an excellent sensitivity and is able to detect false-negative results in immunofluorescence, which occur in a very small percentage (0.043%). The disadvantage of this confirmatory assay is the sacrification of relatively high numbers of mice (in the Swiss rabies center about 1,300 animals each year), and the long time required for a final diagnosis: 7-20 days in positive, 21 days in negative cases. The cultivation of virus from brain suspensions on a mouse neuroblastoma cell line is a tempting alternative to the mouse inoculation test. This method usually provides a conclusive diagnosis within a few days. However, in our hands it showed in preliminary experiments an unsatisfactory sensitivity (80.7%). The necessity to carry out strict reproducibility controls in this assay has to be emphasized. Further work must be invested in the improvement of the rabies tissue culture infection test and a careful long-term comparison with the mouse inoculation test will be necessary before the mouse inoculation test can be replaced.

  8. Carbapenem-resistant Enterobacteriaceae: frequency of hospital room contamination and survival on various inoculated surfaces.

    Science.gov (United States)

    Weber, David J; Rutala, William A; Kanamori, Hajime; Gergen, Maria F; Sickbert-Bennett, Emily E

    2015-05-01

    Carbapenem-resistant Enterobacteriaceae (CRE) only contaminated the environmental surfaces of rooms housing CRE colonized/infected patients infrequently (8.4%) and at low levels (average, 5.1 colony-forming units [CFU]/120 cm² per contaminated surface). Three species of CRE (Klebsiella, Enterobacter, and Escherichia) survived poorly (>85% die-off in 24 hours) when ~2 log10 CFU were inoculated onto 5 different environmental surfaces.

  9. Efficacy of alkaline washing for the decontamination of orange fruit surfaces inoculated with Escherichia coli.

    Science.gov (United States)

    Pao, S; Davis, C L; Kelsey, D F

    2000-07-01

    The effectiveness of washing treatments to decontaminate orange fruit surfaces inoculated with Escherichia coli was evaluated. Washing on roller brushes with fruit cleaners or sanitizers followed by potable water rinse reduced E. coli by 1.9 to 3.5 log cycles. Prewetting fruit for 30 s before washing provided no significant benefit in most cases. Additional sanitizing treatments either with chlorine or acid sanitizers did not enhance the results of alkaline washing. In general, high pH washing solutions (pH 11.8) applied with an adequate spray volume effectively reduced the surface contamination of fruit that lowered the microbial load of fresh juice as well.

  10. Development and hydrology of biological soil crusts -- first results from a surface inoculation experiment

    Science.gov (United States)

    Mykhailova, Larysa; Raab, Thomas; Gypser, Stella; Fischer, Thomas

    2016-04-01

    Representing a set of various micro-biocoenoses, biocrusts often reside in adjacent patches, which not necessarily relate to structural elements of the habitat, like (micro-) topography or vegetational patterns. Such biocrust patches may become more stable through the formation of mutually dependent ecohydrological regimes. For example, algal patches inhibiting infiltration and generating runoff alternate with runoff-receiving moss patches possessing high water holding capacities. Here, we preliminarily report on a lysimeter field experiment where natural biocrust isolates were used for surface inoculation to (I) prove stochastic vs. deterministic biocrust development and (II) to quantitatively relate biocrust development to soil hydrology. Lysimeter sand was collected from 3-4 m below surface at natural dune outcrops in south-eastern Brandenburg, Germany (Glashütte (GLA) and Neuer Lugteich (LUG)), where biocrust samples were collected at the respective dune bases. The lysimeters were designed to prevent runoff. In a completely randomized full-factorial design, three factors were considered. (A) Inocolum in three treatments (bare control, mosses, algae), (B) mineral substrate texture in two treatments (GLA: 55% and LUG: 79% particles >630 μm), and (C) surface compaction in two treatments (control, 41.5 kN m-2 for 30 seconds). The samples were kept dry and re-moistened to -60 hPa two days before inoculation. After a species inventory, the inoculate was isolated by gently washing off sand particles from the biocrust samples. Algal/lichen crusts were dominated by Zygogonium ericetorum and Cladonia sp. at both sites. All moss crusts were dominated by Polytrichum piliferum and Ceratodon purpureus, whereas Brachythecium albicans was present at GLA only. 20 g of homogenized moist inoculate were spread over the surface of each lysimeter (Ø 19 cm, 22 cm depth). We performed autochthonous inoculation, i.e. biocrust isolates collected from GLA were used for inoculation of

  11. Lack of prion accumulation in lymphoid tissues of PRNP ARQ/ARR sheep intracranially inoculated with the agent of scrapie.

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    Greenlee, Justin J; Kunkle, Robert A; Richt, Jürgen A; Nicholson, Eric M; Hamir, Amir N

    2014-01-01

    Sheep scrapie is a transmissible spongiform encephalopathy that can be transmitted horizontally. The prion protein gene (PRNP) profoundly influences the susceptibility of sheep to the scrapie agent and the tissue levels and distribution of PrPSc in affected sheep. The purpose of this study was to compare the survival time and PrPSc tissue distribution in sheep with highly resistant and highly susceptible PRNP genotypes after intracranial inoculation of the agent of scrapie. Five sheep each of genotype VRQ/VRQ, VRQ/ARR or ARQ/ARR were inoculated. Sheep were euthanized when clinical signs of scrapie became severe. Clinical signs, microscopic lesions, and western blot profiles were uniform across genotypes and consistent with manifestations of classical scrapie. Mean survival time differences were associated with the 171 polymorphic site with VRQ/VRQ sheep surviving 18 months, whereas VRQ/ARR and ARQ/ARR sheep survived 60 and 56 months, respectively. Labeling of PrPSc by immunohistochemistry revealed similar accumulations in central nervous system tissues regardless of host genotype. Immunoreactivity for PrPSc in lymphoid tissue was consistently abundant in VRQ/VRQ, present but confined to tonsil or retropharyngeal lymph node in 4/5 VRQ/ARR, and totally absent in ARQ/ARR sheep. The results of this study demonstrate the susceptibility of sheep with the ARQ/ARR genotype to scrapie by the intracranial inoculation route with PrPSc accumulation in CNS tissues, but prolonged incubation times and lack of PrPSc in lymphoid tissue.

  12. Pharmacokinetics and tissue distribution of amoxicillin in healthy and Salmonella Typhimurium-inoculated pigs

    DEFF Research Database (Denmark)

    Agerso, H.; Friis, C.; Nielsen, Jens

    2000-01-01

    chromatography 4, 8, 12, and 24 hours after IM administration. Pharmacokinetic values of amoxicillin in plasma were assessed by use of a l-compartment model with first-order absorption. Results--Inoculation caused diarrhea and increased rectal temperature and WBC count. Absorption half-life was shorter...

  13. Effect of inoculum size, bacterial species, type of surfaces and contact time to the transfer of foodborne pathogens from inoculated to non-inoculated beef fillets via food processing surfaces.

    Science.gov (United States)

    Gkana, E; Chorianopoulos, N; Grounta, A; Koutsoumanis, K; Nychas, G-J E

    2017-04-01

    The objective of the present study was to determine the factors affecting the transfer of foodborne pathogens from inoculated beef fillets to non-inoculated ones, through food processing surfaces. Three different levels of inoculation of beef fillets surface were prepared: a high one of approximately 10(7) CFU/cm(2), a medium one of 10(5) CFU/cm(2) and a low one of 10(3) CFU/cm(2), using mixed-strains of Listeria monocytogenes, or Salmonella enterica Typhimurium, or Escherichia coli O157:H7. The inoculated fillets were then placed on 3 different types of surfaces (stainless steel-SS, polyethylene-PE and wood-WD), for 1 or 15 min. Subsequently, these fillets were removed from the cutting boards and six sequential non-inoculated fillets were placed on the same surfaces for the same period of time. All non-inoculated fillets were contaminated with a progressive reduction trend of each pathogen's population level from the inoculated fillets to the sixth non-inoculated ones that got in contact with the surfaces, and regardless the initial inoculum, a reduction of approximately 2 log CFU/g between inoculated and 1st non-inoculated fillet was observed. S. Typhimurium was transferred at lower mean population (2.39 log CFU/g) to contaminated fillets than E. coli O157:H7 (2.93 log CFU/g), followed by L. monocytogenes (3.12 log CFU/g; P CFU/g) enhanced the transfer of bacteria to subsequent fillets compared to other materials (2.66 log CFU/g for SS and PE; P < 0.05). Cross-contamination between meat and surfaces is a multifactorial process strongly depended on the species, initial contamination level, kind of surface, contact time and the number of subsequent fillet, according to analysis of variance. Thus, quantifying the cross-contamination risk associated with various steps of meat processing and food establishments or households can provide a scientific basis for risk management of such products.

  14. Influence of Rhizobia Inoculation on Biomass Gain and Tissue Nitrogen Content of Leucaena leucocephala Seedlings under Drought

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    Gabriela Pereyra

    2015-10-01

    Full Text Available Anticipated increases in the frequency of heat waves and drought spells may have negative effects on the ability of leguminous trees to fix nitrogen (N. In seedlings of Leucaena leucocephala inoculated with Mesorhizobium loti or Rhizobium tropici, we investigated how the developmental stage and a short drought influenced overall biomass and the accumulation of carbon and N in plant tissues. In early developmental stages, the number of nodules and nodule biomass were correlated with total plant biomass and δ15N, and nodules and roots contributed 33%–35% of the seedling total N. Seedlings associated with R. tropici fixed more N and exhibited higher overall biomass compared with M. loti seedlings. Four and a half months after inoculation (140 days after inoculation, DAI, a short (15-day drought inhibited seedling growth and caused a decline in total plant N, with the smallest decline in R. tropici seedlings. After 15 days of drought, i.e., 155 DAI, the nodules had accumulated proline, but the total amino acid concentration did not change. Our results indicate that N-fixation is independent of seedlings growth. In addition, R. tropici is a better choice than M. loti as a symbiont for Leucaena seedlings for forest restoration and agroforestry applications under increasingly drier conditions.

  15. Differential expression of genes identified from Poncirus trifoliata tissue inoculated with CTV through EST analysis and in silico hybridization

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    Mariângela Cristofani-Yaly

    2007-01-01

    Full Text Available Citrus is the most important fruit crop in Brazil and Citrus tristeza virus (CTV is considered one of the most important pathogens of citrus. Most citrus species and varieties are susceptible to CTV infection. However, Poncirus trifoliata, a close relative of citrus, is resistant to the virus. In order to better understand the responses of citrus plants to the infection of CTV, we constructed expressed sequence tag (EST libraries with tissues collected from Poncirus trifoliata plants, inoculated or not with Citrus tristeza virus at 90 days after inoculation, grafted on Rangpur lime rootstocks. We generated 17,867 sequence tags from Poncirus trifoliata inoculated (8,926 reads and not (8,941 reads with a severe CTV isolate. A total of 2,782 TCs (Tentative Consensi sequences were obtained using both cDNA libraries in a single clusterization procedure. By the in silico hybridization approach, 289 TCs were identified as differentially expressed in the two libraries. A total of 121 TCs were found to be overexpressed in plants infected with CTV and were grouped in 12 primary functional categories. The majority of them were associated with metabolism and defense response. Some others were related to lignin, ethylene biosynthesis and PR proteins. In general, the differentially expressed transcripts seem to be somehow involved in secondary plant response to CTV infection.

  16. Evaluation of inoculation method and inoculum drying time for their effects on survival and efficiency of recovery of Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes inoculated on the surface of tomatoes.

    Science.gov (United States)

    Lang, Megan M; Harris, Linda J; Beuchat, Larry R

    2004-04-01

    A study was undertaken to evaluate methods for applying inoculum and to examine the effect of inoculum drying time on survival and recovery of foodborne pathogens inoculated onto the surface of raw, ripe tomatoes. Five-strain mixtures of Escherichia coli O157:H7, Salmonella, or Listeria monocytogenes were applied to tomatoes by dip, spot, or spray inoculation methods. Inocula were dried for 1 or 24 h at 22 degrees C before tomatoes were treated with water (control) or chlorine (200 micrograms/ml). Significantly (alpha = 0.05) larger populations (CFU per tomato) of E. coli O157:H7 and Salmonella were recovered from dipinoculated tomatoes than from spot- or spray-inoculated tomatoes. This difference was attributed to larger numbers of cells adhering to tomatoes subjected to dip inoculation. Populations of E. coli O157:H7 and Salmonella recovered from spot- and spray-inoculated tomatoes containing the same initial number of cells were not significantly different. Significantly different L. monocytogenes population sizes were recovered from inoculated tomatoes (dip > spot > spray). Populations of pathogens recovered from tomatoes were significantly larger when inocula were dried for 1 h compared with 24 h. Significant differences (water > chlorine) were observed in the sizes of populations for all pathogens recovered from tomatoes treated with chlorine, regardless of inoculation method or drying time. Results indicate that inoculation method, drying time, and treatment affect survival and/or recovery of foodborne pathogens inoculated onto the surface of tomatoes. We recommend that spot inoculation with a drying time of 24 h at 22 degrees C be used with standard methods to determine the efficacy of chlorine and other sanitizers for killing foodborne pathogens on tomatoes.

  17. A novel model to assess the efficacy of steam surface pasteurization of cooked surimi gels inoculated with realistic levels of Listeria innocua.

    Science.gov (United States)

    Skåra, Torstein; Valdramidis, Vasilis P; Rosnes, Jan Thomas; Noriega, Estefanía; Van Impe, Jan F M

    2014-12-01

    Steam surface pasteurization is a promising decontamination technology for reducing pathogenic bacteria in different stages of food production. The effect of the artificial inoculation type and initial microbial load, however, has not been thoroughly assessed in the context of inactivation studies. In order to optimize the efficacy of the technology, the aim of this study was to design and validate a model system for steam surface pasteurization, assessing different inoculation methods and realistic microbial levels. More specifically, the response of Listeria innocua, a surrogate organism of Listeria monocytogenes, on a model fish product, and the effect of different inoculation levels following treatments with a steam surface pasteurization system was investigated. The variation in the resulting inoculation level on the samples was too large (77%) for the contact inoculation procedure to be further considered. In contrast, the variation of a drop inoculation procedure was 17%. Inoculation with high levels showed a rapid 1-2 log decrease after 3-5 s, and then no further inactivation beyond 20 s. A low level inoculation study was performed by analysing the treated samples using a novel contact plating approach, which can be performed without sample homogenization and dilution. Using logistic regression, results from this method were used to model the binary responses of Listeria on surfaces with realistic inoculation levels. According to this model, a treatment time of 23 s will result in a 1 log reduction (for P = 0.1).

  18. Recovery and Behaviour of Stressed Escherichia coli O157:H7 Cells on Rocket Leaf Surfaces Inoculated by Different Methods

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    ANAS A. AL-NABULSI

    2016-03-01

    Full Text Available E. coli O157:H7 is an emerging public health concern worldwide because of its low infectious dose and ability to survive under adverse conditions. Tests were conducted to determine the abilityof unstressed E. coli O157:H7 cells or those stressed by acid, cold, salt exposure or starvation to survive on the surfaces of rocket leaves after contamination by three methods (dip, spray or spot inoculation and following storage at 10 or 25ºC. E. coli O157:H7 numbers recovered from rocket leaves contaminated by the different techniques were in the order of dip > spot > spray inoculation.Numbers of stressed E. coli O157:H7 recovered after inoculation by all three methods increased significantly over 7d storage at 10 or 25ºC, while unstressed E. coli O157:H7 only grew following dip inoculation. Exposure to adverse environmental conditions may increase the risk of E. coli O157:H7 survival and spread on leafy green vegetables.

  19. Virus expression in different tissues of normal and tumor-bearing mice inoculated with a murine leukemia virus.

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    Youn, J K; Santillana, M; Hue, G; Barski, G

    1977-11-15

    Evolution of virus expression in different lymphoid organs as well as in solid syngeneic tumors of mice inoculated with an MuLV was studied with the aid of in vitro XC co-culture technique. When normal adult mice of strain XLII were inoculated intraperitoneally with a cultured Rauscher virus (RC), the virus could be detected, 10 days after inoculation, only in bone marrow in small amounts and thereafter no virus could be found in any of the organs tested, including bone marrow, spleen, thymus, lymph node and kidney. However, when age- and sex-matched parallel mice bearing syngeneic subcutaneous non-viral tumors were inoculated similarly with the RC virus, the virus could be detected abundantly not only in bone marrow and spleen but also in tumors during the first 3 weeks and even 6 weeks after virus inoculation. Transitional decrease or disappearance of the virus was observed around the 25th-31st day in organs and tumors of the inoculated mice. When the tumor mass was removed from these mice by surgery, the virus disappeared rapidly and definitely from all the organs tested. The virus recovered from in vitro explanted and cultured tumors, taken from mice inoculated with the virus, induced typical lymphoid leukemia in BALB/c mice inoculated as newborns. However, from certain aspects (hypertrophy of the thymus and lymph nodes), this virus was different from the original RC virus.

  20. Hypertrophy, hyperplasia, and infectious virus in gut-associated lymphoid tissue of mice after oral inoculation with simian-human or bovine-human reassortant rotaviruses.

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    Moser, C A; Dolfi, D V; Di Vietro, M L; Heaton, P A; Offit, P A; Clark, H F

    2001-04-01

    Oral inoculation of infants with a vaccine that contains simian-human reassortant rotaviruses has been found to be a rare cause of intussusception. Because intussusception can be associated with enlargement of gut-associated lymphoid tissue, we studied the capacity of simian-human and bovine-human reassortant rotaviruses to cause lymphoid hypertrophy and hyperplasia of Peyer's patches (PP) of adult BALB/c mice. Neither hypertrophy nor hyperplasia was detected in PP after oral inoculation with simian-human or bovine-human reassortant rotaviruses. However, infectious virus was detected in PP and mesenteric lymph nodes after oral inoculation with simian, but not bovine, reassortant rotaviruses. Implications of these findings on the pathogenesis of intussusception are discussed.

  1. Surface modulation of dental hard tissues

    Science.gov (United States)

    Tantbirojn, Daranee

    Tooth surfaces play a central role in the equilibrium of dental hard tissues, in which contrasting processes lead to loss or deposition of materials. The central interest of this Thesis was the modulation of tooth surfaces to control such equilibrium. Four specific studies were carried out to investigate different classes of surface modulating agents. These are: (1) Ionic modulation of the enamel surface to enhance stain removal . Dental stain is the most apparent form of tooth surface deposit. The nature of extrinsic stain in terms of spatial chemical composition was studied by using electron probe microanalysis. An ionic surface modulating agent, sodium tripolyphosphate (STPP), was evaluated. Image analysis methodologies were developed and the ability of STPP in stain removal was proved. (2) Thin film modulation with substantive polymeric coating and the effect on in vitro enamel de/re-mineralization . A novel polymeric coating that formed a thin film on the tooth surface was investigated for its inhibitory effect on artificial enamel caries, without interfering with the remineralization process. The preventive effect was distinct, but the mineral redeposition was questionable. (3) Thick film modulation with fluoride containing sealants and the effect on in vitro enamel and root caries development. Fluoride incorporated into resin material is an example of combining different classes of surface modulating agents to achieve an optimal outcome. A proper combination, such as in resin modified glass ionomer, showed in vitro caries inhibitory effect beyond the material boundary in both enamel and dentin. (4) Thick film modulation with dental adhesives and the determination of adhesion to dentin. Dentin adhesives modulate intracoronal tooth surfaces by enhancing adhesion to restorative materials. Conventional nominal bond tests were inadequate to determine the performance of current high strength adhesives. It was shown that interfacial fracture toughness test was more

  2. Homeostatic fluctuations of a tissue surface

    CERN Document Server

    Risler, Thomas; Prost, Jacques

    2015-01-01

    We study the surface fluctuations of a tissue with a dynamics dictated by cell-rearrangement, cell-division and cell-death processes. Surface fluctuations are calculated in the homeostatic state, where cell division and cell death equilibrate on average. The obtained fluctuation spectrum can be mapped onto several other spectra such as those characterizing incompressible fluids, compressible Maxwell elastomers or permeable membranes in appropriate asymptotic regimes. Since cell division and cell death are out-of-equilibrium processes, detailed balance is broken, but a generalized fluctuation-response relation is satisfied in terms of appropriate observables. Our work is a first step toward the description of the out-of-equilibrium fluctuations of the surface of a thick epithelium and its dynamical response to external perturbations.

  3. Tissue localization, shedding, virus carriage, antibody response, and aerosol transmission of Porcine epidemic diarrhea virus following inoculation of 4-week-old feeder pigs.

    Science.gov (United States)

    Niederwerder, Megan C; Nietfeld, Jerome C; Bai, Jianfa; Peddireddi, Lalitha; Breazeale, Barbara; Anderson, Joe; Kerrigan, Maureen A; An, Baoyan; Oberst, Richard D; Crawford, Kimberly; Lager, Kelly M; Madson, Darin M; Rowland, Raymond R R; Anderson, Gary A; Hesse, Richard A

    2016-11-01

    We determined tissue localization, shedding patterns, virus carriage, antibody response, and aerosol transmission of Porcine epidemic diarrhea virus (PEDV) following inoculation of 4-week-old feeder pigs. Thirty-three pigs were randomly assigned to 1 of 3 groups for the 42-day study: inoculated (group A; n = 23), contact transmission (group B; n = 5), and aerosol transmission (group C; n = 5). Contact transmission occurred rapidly to group B pigs whereas productive aerosol transmission failed to occur to group C pigs. Emesis was the first clinical sign noted at 3 days postinoculation (dpi) followed by mild to moderate diarrhea lasting 5 more days. Real-time PCR detected PEDV in fecal and nasal swabs, oral fluids, serum, and gastrointestinal and lymphoid tissues. Shedding occurred primarily during the first 2 weeks postinoculation, peaking at 5-6 dpi; however, some pigs had PEDV nucleic acid detected in swabs collected at 21 and 28 dpi. Antibody titers were measurable between 14 and 42 dpi. Although feces and intestines collected at 42 dpi were PEDV negative by PCR and immunohistochemistry, respectively, small intestines from 70% of group A pigs were PCR positive. Although disease was relatively mild and transient in this age group, the results demonstrate that 4-week-old pigs are productively infected and can sustain virus replication for several weeks. Long-term shedding of PEDV in subclinically affected pigs should be considered an important source for PEDV transmission.

  4. Transfer of Pathogens from Cantaloupe Rind to Preparation Surfaces and Edible Tissue as a Function of Cutting Method.

    Science.gov (United States)

    Shearer, Adrienne E H; LeStrange, Kyle; Castañeda Saldaña, Rafael; Kniel, Kalmia E

    2016-05-01

    Whole and cut cantaloupes have been implicated as vehicles in foodborne illness outbreaks of norovirus, salmonellosis, and listeriosis. Preparation methods that minimize pathogen transfer from external surfaces to the edible tissue are needed. Two preparation methods were compared for the transfer of Listeria monocytogenes, Salmonella enterica serovar Typhimurium LT2, murine norovirus, and Tulane virus from inoculated cantaloupe rinds to edible tissue and preparation surfaces. For the first method, cantaloupes were cut into eighths, and edible tissue was separated from the rind and cubed with the same knife used to open the cantaloupes. For the second method, cantaloupes were scored with a knife around the circumference sufficient to allow manual separation of the cantaloupes into halves. Edible tissue was scooped with a spoon and did not contact the preparation surface touched by the rind. Bacteria and virus were recovered from the rinds, preparation surfaces, and edible tissue and enumerated by culture methods and reverse transcription, quantitative PCR, respectively. Standard plate counts were determined throughout refrigerated storage of cantaloupe tissue. Cut method 2 yielded approximately 1 log lower recovery of L. monocytogenes and Salmonella Typhimurium from edible tissue, depending on the medium in which the bacteria were inoculated. A slight reduction was observed in murine norovirus recovered from edible tissue by cut method 2. The Tulane virus was detected in approximately half of the sampled cantaloupe tissue and only at very low levels. Aerobic mesophilic colony counts were lower through day 6 of storage for buffered peptone water-inoculated cantaloupes prepared by cut method 2. No differences were observed in environmental contamination as a function of cutting method. Although small reductions in contamination of edible tissue were observed for cut method 2, the extent of microbial transfer underscores the importance of preventing contamination of

  5. Effect of Morphology of Instantaneous Inoculant on Inoculated Result of Melted Iron

    Institute of Scientific and Technical Information of China (English)

    SHENG Da

    2003-01-01

    The characteristics of some elements in inoculant were analyzed. The effect of the morphology of instantaneous inoculant on its melting velocity was studied. When the inoculants pass through the same sieve number, the volume and the ratio of surface area to volume are different. It is evident from the theoretical analysis and experiment under some conditions that the melting velocity of inoculant depends on the morphology of inoculant. The morphology of inoculant during production should be controlled carefully.

  6. Cytokine and acute phase protein mRNA expression in liver tissue from pigs with severe sepsis caused by intravenous inoculation of Staphylococcus aureus

    DEFF Research Database (Denmark)

    Nielsen, Ole Lerberg; Olsen, Helle Gerda; Iburg, Tine

    2010-01-01

    The aim was to substantiate previous findings of hepatic dysfunction in a porcine model of Staphylococcus aureus induced severe sepsis. Nine pigs were inoculated intravenously once or twice with 108S. aureus per kilogram body weight and killed 12, 24 and 48 h later. Three pigs served as controls....... Blood was sampled for bacteriology, haematology and clinical pathology. Tissues were collected at necropsy for bacteriology, gene expression analysis and histology. Bacterial counts in blood remained low, decreased in the lungs, liver and spleen, but increased in bone. All infected pigs developed sepsis...... of IL-6, IL-8, IL-1β, and CRP. N o increase could be detected in the IL-1α or TNFα liver-mRNA levels. IL-6, IL-8 and IL-1β expression peaked at 24 hours (2-5 fold compared to the control group). In conclusion, the increased liver cytokine mRNA levels indicate a local hepatic, non-infectious inflammatory...

  7. Inoculation of sugarcane with Pantoea sp. increases amino acid contents in shoot tissues; serine, alanine, glutamine and asparagine permit concomitantly ammonium excretion and nitrogenase activity of the bacterium.

    Science.gov (United States)

    Loiret, F G; Grimm, B; Hajirezaei, M R; Kleiner, D; Ortega, E

    2009-07-15

    Pantoea sp. is an endophytic nitrogen-fixing bacterium isolated from sugarcane tissues. The aim of the present study was to determine the contents of amino acids in sugarcane as a result of inoculation of nodes and nodal roots with Pantoea sp. strain 9C and to evaluate the effects of amino acids on growth, nitrogenase activity and ammonium excretion of the bacterium. Content of almost all amino acids increased in 30-day-old plantlets by root inoculation. The most abundant amino acids in shoot tissues were asparagine and proline, and those in nodal roots were asparagine, proline, aspartic acid, glutamic acid and serine. The bacterium was able to grow on all tested amino acids except histidine, isoleucine and leucine. Nitrogenase Pantoea sp. was partially inhibited by 1, 2 or 5mmolL(-1) and completely inhibited by 10mmolL(-1) of NH(4)(+) in the media. Pantoea sp. showed nitrogenase activity in 5mmolL(-1) of serine, asparagine, threonine, alanine, proline, tyrosine, valine, methionine, lysine, phenylalanine, cysteine, tryptophan, citrulline and ornithine. Pantoea sp. did not excrete ammonium when it grew in vivo conditions favoring nitrogen fixation; however, ammonium was detected in the supernatant when 5mmolL(-1) asparagine, aspartic acid, alanine, serine or glutamine was added to the medium. The highest ammonium concentration in the supernatant was detected, when Pantoea grew on serine. Ammonium in the supernatant and nitrogenase activity were only detectable concomitantly when the medium was supplemented with serine, alanine, glutamine or asparagine. We discuss roles of amino acids on plant-bacteria interaction during the colonization of sugarcane plants.

  8. Surface modification of polyester biomaterials for tissue engineering.

    Science.gov (United States)

    Jiao, Yan-Peng; Cui, Fu-Zhai

    2007-12-01

    Surfaces play an important role in a biological system for most biological reactions occurring at surfaces and interfaces. The development of biomaterials for tissue engineering is to create perfect surfaces which can provoke specific cellular responses and direct new tissue regeneration. The improvement in biocompatibility of biomaterials for tissue engineering by directed surface modification is an important contribution to biomaterials development. Among many biomaterials used for tissue engineering, polyesters have been well documented for their excellent biodegradability, biocompatibility and nontoxicity. However, poor hydrophilicity and the lack of natural recognition sites on the surface of polyesters have greatly limited their further application in the tissue engineering field. Therefore, how to introduce functional groups or molecules to polyester surfaces, which ideally adjust cell/tissue biological functions, becomes more and more important. In this review, recent advances in polyester surface modification and their applications are reviewed. The development of new technologies or methods used to modify polyester surfaces for developing their biocompatibility is introduced. The results of polyester surface modifications by surface morphological modification, surface chemical group/charge modification, surface biomacromolecule modification and so on are reported in detail. Modified surface properties of polyesters directly related to in vitro/vivo biological performances are presented as well, such as protein adsorption, cell attachment and growth and tissue response. Lastly, the prospect of polyester surface modification is discussed, especially the current conception of biomimetic and molecular recognition.

  9. Interdigitated interdigital transducer for surface elastometry of soft damping tissue.

    Science.gov (United States)

    Danicki, Eugene; Nowicki, Andrzej; Tasinkevych, Yuriy

    2013-06-01

    Measurement of the shear elastic constant of soft and highly damping tissue of high Poisson ratio is quite a challenging task. It is proposed to evaluate shear wave velocity and damping of tissue by measuring the shear skimming bulk waves using one interdigitated interdigital transducer on a piezoelectric layer, such as polyvinylidene fluoride, applied to the surface of the small tissue sample.

  10. Laser microtexturing of implant surfaces for enhanced tissue integration

    Energy Technology Data Exchange (ETDEWEB)

    Ricci, J.L. [Univ. of Medicine and Dentistry of New Jersey, Newark, NJ (United States). Dept. of Orthodontics; Alexander, H. [Orthogen Corp., Springfield, NJ (United States)

    2001-07-01

    The success or failure of bone and soft tissue-fixed medical devices, such as dental and orthopaedic implants, depends on a complex combination of biological and mechanical factors. These factors are intimately associated with the interface between the implant surface and the surrounding tissue, and are largely determined by the composition, surface chemistry, and surface microgeometry of the implant. The relative contributions of these factors are difficult to assess. This study addresses the contribution of surface microtexture, on a controlled level, to tissue integration. (orig.)

  11. Probiotic table olives: microbial populations adhering on olive surface in fermentation sets inoculated with the probiotic strain Lactobacillus paracasei IMPC2.1 in an industrial plant.

    Science.gov (United States)

    De Bellis, Palmira; Valerio, Francesca; Sisto, Angelo; Lonigro, Stella Lisa; Lavermicocca, Paola

    2010-05-30

    This study reports the dynamics of microbial populations adhering on the surface of debittered green olives cv. Bella di Cerignola in fermentation sets inoculated with the probiotic strain Lactobacillus paracasei IMPC2.1 in different brining conditions (4% and 8% (w/v) NaCl) at room temperature and 4 degrees C. The probiotic strain successfully colonized the olive surface dominating the natural LAB population and decreasing the pH of brines to fermentation. The dynamics of microbial populations associated with olive surface and belonging to the different groups indicated that inoculated olives held at room temperature did not host Enterobacteriaceae at the end of fermentation. Yeast populations were present in a low number (fermentation in all processes except for the one held at 4 degrees C. Also a notable incidence of Leuc. mesenteroides on olives was highlighted in this study during all fermentation. Results indicated that the human strain L. paracasei IMPC2.1 can be considered an example of a strain used in the dual role of starter and probiotic culture which allowed the control of fermentation processes and the realization of a final probiotic product with functional appeal.

  12. A skull stripping method using deformable surface and tissue classification

    Science.gov (United States)

    Tao, Xiaodong; Chang, Ming-Ching

    2010-03-01

    Many neuroimaging applications require an initial step of skull stripping to extract the cerebrum, cerebellum, and brain stem. We approach this problem by combining deformable surface models and a fuzzy tissue classification technique. Our assumption is that contrast exists between brain tissue (gray matter and white matter) and cerebrospinal fluid, which separates the brain from the extra-cranial tissue. We first analyze the intensity of the entire image to find an approximate centroid of the brain and initialize an ellipsoidal surface around it. We then perform a fuzzy tissue classification with bias field correction within the surface. Tissue classification and bias field are extrapolated to the entire image. The surface iteratively deforms under a force field computed from the tissue classification and the surface smoothness. Because of the bias field correction and tissue classification, the proposed algorithm depends less on particular imaging contrast and is robust to inhomogeneous intensity often observed in magnetic resonance images. We tested the algorithm on all T1 weighted images in the OASIS database, which includes skull stripping results using Brain Extraction Tool; the Dice scores have an average of 0.948 with a standard deviation of 0.017, indicating a high degree of agreement. The algorithm takes on average 2 minutes to run on a typical PC and produces a brain mask and membership functions for gray matter, white matter, and cerebrospinal fluid. We also tested the algorithm on T2 images to demonstrate its generality, where the same algorithm without parameter adjustment gives satisfactory results.

  13. Measurement of aggregate cohesion by tissue surface tensiometry.

    Science.gov (United States)

    Butler, Christine M; Foty, Ramsey A

    2011-04-08

    Rigorous measurement of intercellular binding energy can only be made using methods grounded in thermodynamic principles in systems at equilibrium. We have developed tissue surface tensiometry (TST) specifically to measure the surface free energy of interaction between cells. The biophysical concepts underlying TST have been previously described in detail. The method is based on the observation that mutually cohesive cells, if maintained in shaking culture, will spontaneously assemble into clusters. Over time, these clusters will round up to form spheres. This rounding-up behavior mimics the behavior characteristic of liquid systems. Intercellular binding energy is measured by compressing spherical aggregates between parallel plates in a custom-designed tissue surface tensiometer. The same mathematical equation used to measure the surface tension of a liquid droplet is used to measure surface tension of 3D tissue-like spherical aggregates. The cellular equivalent of liquid surface tension is intercellular binding energy, or more generally, tissue cohesivity. Previous studies from our laboratory have shown that tissue surface tension (1) predicts how two groups of embryonic cells will interact with one another, (2) can strongly influence the ability of tissues to interact with biomaterials, (3) can be altered not only through direct manipulation of cadherin-based intercellular cohesion, but also by manipulation of key ECM molecules such as FN and 4) correlates with invasive potential of lung cancer, fibrosarcoma, brain tumor and prostate tumor cell lines. In this article we will describe the apparatus, detail the steps required to generate spheroids, to load the spheroids into the tensiometer chamber, to initiate aggregate compression, and to analyze and validate the tissue surface tension measurements generated.

  14. Effects of silicon treatment and inoculation with Fusarium oxysporum f. sp. vasinfectum on cellular defences in root tissues of two cotton cultivars.

    Science.gov (United States)

    Whan, Jennifer A; Dann, Elizabeth K; Aitken, Elizabeth A B

    2016-08-01

    Silicon has been shown to enhance the resistance of plants to fungal and bacterial pathogens. Here, the effect of potassium silicate was assessed on two cotton (Gossypium hirsutum) cultivars subsequently inoculated with Fusarium oxysporum f. sp. vasinfectum (Fov). Sicot 189 is moderately resistant whilst Sicot F-1 is the second most resistant commercial cultivar presently available in Australia. Transmission and light microscopy were used to compare cellular modifications in root cells after these different treatments. The accumulation of phenolic compounds and lignin was measured. Cellular alterations including the deposition of electron-dense material, degradation of fungal hyphae and occlusion of endodermal cells were more rapidly induced and more intense in endodermal and vascular regions of Sicot F-1 plants supplied with potassium silicate followed by inoculation with Fov than in similarly treated Sicot 189 plants or in silicate-treated plants of either cultivar not inoculated with Fov. Significantly more phenolic compounds were present at 7 d post-infection (dpi) in root extracts of Sicot F-1 plants treated with potassium silicate followed by inoculation with Fov compared with plants from all other treatments. The lignin concentration at 3 dpi in root material from Sicot F-1 treated with potassium silicate and inoculated with Fov was significantly higher than that from water-treated and inoculated plants. This study demonstrates that silicon treatment can affect cellular defence responses in cotton roots subsequently inoculated with Fov, particularly in Sicot F-1, a cultivar with greater inherent resistance to this pathogen. This suggests that silicon may interact with or initiate defence pathways faster in this cultivar than in the less resistant cultivar. © The Author 2016. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  15. Role of viscosity and surface tension of zebrafish embryonic tissues in tissue flows during gastrulation.

    Science.gov (United States)

    Schoetz, E. M.; Bacarian, T.; Steinberg, M. S.; Burdine, R. D.; Bialek, W.; Heisenberg, C. P.; Foty, R. A.; Julicher, F.

    2007-03-01

    At the onset of gastrulation in zebrafish, complex flows and cell movements occur, which are not well understood. Here, we study the material properties of zebrafish embryonic tissues which are important for the tissue dynamics. We found that these tissues behave viscoelastic and exhibit liquid-like properties on long time scales. They relax internal stress caused by compressive forces or, in the absence of external forces, round up and fuse into spheres to minimize their free surface. Quantitative differences in the adhesivity between different types of tissues result in their immiscibility and sorting behavior analogous to that of ordinary immiscible liquids. When mixed, cells segregate into discrete phases, and the position adopted correlates with differences in the aggregate surface tensions for these phases. Surface tensions were measured with a tissue surface tensiometer. Aggregates were compressed and their force response and shape were recorded as a function of time. From the analysis of the force-relaxation curves, we determined the surface tensions, relaxation times, tissue viscosities and shear moduli. Furthermore, by 4D-cell tracking, we measured kinetic parameters such as cell speed, directionality and persistence of cell movement.

  16. Feasibility of detecting aflatoxin B1 on inoculated maize kernels surface using Vis/NIR hyperspectral imaging

    Science.gov (United States)

    The feasibility of using a visible/near-infrared hyperspectral imaging system with a wavelength range between 400 and 1000 nm to detect and differentiate different levels of aflatoxin B1 (AFB1) artificially titrated on maize kernel surface was examined. To reduce the color effects of maize kernels, ...

  17. Comparison of connective tissue graft and guided tissue regeneration in covering root surfaces

    OpenAIRE

    LotfazarM.; Faghihi Sh.

    2002-01-01

    There are many researches evaluation different methods for covering the root surface. In the most of these studies, type I and II of Miller treatment had been searched. The purpose of this study was a comparison between connective tissue graft (CTG) and guided tissue regeneration (GTR) with a collagen membrane in the treatment of gingival recession defects (Miller class III). Six patients, each contributing a pair of Miller class III buccal gingival recessions, were treated. The clinical meas...

  18. Surface and interfacial creases in a bilayer tubular soft tissue

    Science.gov (United States)

    Razavi, Mir Jalil; Pidaparti, Ramana; Wang, Xianqiao

    2016-08-01

    Surface and interfacial creases induced by biological growth are common types of instability in soft biological tissues. This study focuses on the criteria for the onset of surface and interfacial creases as well as their morphological evolution in a growing bilayer soft tube within a confined environment. Critical growth ratios for triggering surface and interfacial creases are investigated both analytically and numerically. Analytical interpretations provide preliminary insights into critical stretches and growth ratios for the onset of instability and formation of both surface and interfacial creases. However, the analytical approach cannot predict the evolution pattern of the model after instability; therefore nonlinear finite element simulations are carried out to replicate the poststability morphological patterns of the structure. Analytical and computational simulation results demonstrate that the initial geometry, growth ratio, and shear modulus ratio of the layers are the most influential factors to control surface and interfacial crease formation in this soft tubular bilayer. The competition between the stretch ratios in the free and interfacial surfaces is one of the key driving factors to determine the location of the first crease initiation. These findings may provide some fundamental understanding in the growth modeling of tubular biological tissues such as esophagi and airways as well as offering useful clues into normal and pathological functions of these tissues.

  19. Modeling disinfection of plastic poultry transport cages inoculated with Salmonella enteritids by slightly acidic electrolyzed water using response surface methodology.

    Science.gov (United States)

    Zang, Y T; Li, B M; Bing, Sh; Cao, W

    2015-09-01

    In order to reduce the risk of enteric pathogens transmission in animal farms, the disinfection effectiveness of slightly acidic electrolyzed water (SAEW, pH 5.85 to 6.53) for inactivating Salmonella Enteritidis on the surface of plastic poultry transport cages was evaluated. The coupled effects of the tap water cleaning time (5 to 15 s), SAEW treatment time (20 to 40 s), and available chlorine concentrations (ACCs) of 30 to 70 mg/l on the reductions of S. Enteritidis on chick cages were investigated using a central composite design of the response surface methodology (RSM). The established RS model had a goodness of fit quantified by the parameter R2 (0.971), as well as a lack of fit test (P>0.05). The maximum reduction of 3.12 log10 CFU/cm2 for S. Enteritidis was obtained for the cage treated with tap water cleaning for 15 s followed by SAEW treatment for 40 s at an ACC of 50 mg/l. Results indicate that the established RS model has shown the potential of SAEW in disinfection of bacteria on cages. © 2015 Poultry Science Association Inc.

  20. A surface wave elastography technique for measuring tissue viscoelastic properties.

    Science.gov (United States)

    Zhang, Xiaoming

    2017-04-01

    A surface wave elastography method is proposed to study the viscoelastic properties of skin by measuring the surface wave speed and attenuation on the skin. Experiments were carried out on porcine skin tissues. The surface wave speed is measured by the change of phase with distance. The wave attenuation is measured by the decay of wave amplitude with distance. The change of viscoelastic properties with temperature was studied at room and body temperatures. The wave speed was 1.83m/s at 22°C but reduced to 1.52m/s at 33°C. The viscoelastic ratio was almost constant from 22°C to 33°C. Fresh and decayed tissues were studied. The wave speed of the decayed tissue increased from 1.83m/s of fresh state to 2.73m/s. The viscoelastic ratio was 0.412/mm at the decayed state compared to 0.215/mm at the fresh state. More tissue samples are needed to study these viscoelastic parameters according to specific applications.

  1. Identification of rheological properties of human body surface tissue.

    Science.gov (United States)

    Benevicius, Vincas; Gaidys, Rimvydas; Ostasevicius, Vytautas; Marozas, Vaidotas

    2014-04-11

    According to World Health Organization obesity is one of the greatest public health challenges of the 21st century. It has tripled since the 1980s and the numbers of those affected continue to rise at an alarming rate, especially among children. There are number of devices that act as a prevention measure to boost person's motivation for physical activity and its levels. The placement of these devices is not restricted thus the measurement errors that appear because of the body rheology, clothes, etc. cannot be eliminated. The main objective of this work is to introduce a tool that can be applied directly to process measured accelerations so human body surface tissue induced errors can be reduced. Both the modeling and experimental techniques are proposed to identify body tissue rheological properties and prelate them to body mass index. Multi-level computational model composed from measurement device model and human body surface tissue rheological model is developed. Human body surface tissue induced inaccuracies can increase the magnitude of measured accelerations up to 34% when accelerations of the magnitude of up to 27 m/s(2) are measured. Although the timeframe of those disruptions are short - up to 0.2 s - they still result in increased overall measurement error.

  2. Under the Surface of Subcutaneous Adipose Tissue Biology.

    Science.gov (United States)

    Pandžić Jakšić, Vlatka; Grizelj, Danijela

    2016-12-01

    The global obesity epidemic enhanced contemporary interest in adipose tissue biology. Two structurally and functionally distinct depots, subcutaneous adipose tissue (SAT) and visceral adipose tissue (VAT), are spread throughout the body. Their distribution was recognized to be a major determinant of metabolic risk. Unlike VAT, SAT showed some protective endocrine and inflammatory features that might explain the occurrence of obese but metabolically healthy persons. The unique developmental gene expression signature, angiogenesis, and adipogenic potential of SAT determines its growth ability under the positive energy balance. The overflow hypothesis suggested that when SAT is unable to expand sufficiently, fat overflows towards metabolically adverse ectopic depots. Besides white adipose tissue, recent studies found important brown adipose tissue activity responsible for thermogenesis and energy dissipation in adults as well. SAT is prone to "browning" - the appearance of particular beige adipocytes that contribute to its favorable metabolic effects. Morbid obesity, aging, hormonal status, nutrition, low physical activity, and other environmental factors impair SAT relative resistance to dysfunctional changes and promote development of metabolic disorders. The popular approach considering SAT mainly as the subject of cosmetic procedures for improving the appearance of body contours should be avoided. Complex heterogeneity of obesity revealed that a tissue of an extreme plasticity and rich interactions with vital functions of the body lies under the surface. Therapeutic manipulations to preserve and enhance healthier fat in order to correct obesity-related metabolic disorders seem to be a relevant but still unexplored opportunity.

  3. Viability of a multi-strain mixture of Listeria monocytogenes, Salmonella typhimurium, or Escherichia coli O157:H7 inoculated into the batter or onto the surface of a soudjouk-style fermented semi-dry sausage

    Science.gov (United States)

    Multi-strain mixtures of Listeria monocytogenes, Salmonella typhimurium, or Escherichia coli O157:H7 were separately inoculated either into soudjouk batter or onto the surface of slices of commercial soudjouk to levels of ca. 6.0 log10 CFU per gram or slice, respectively. After fermentation and dryi...

  4. Surface coating for prevention of metallic seed migration in tissues.

    Science.gov (United States)

    Lee, Hyunseok; Lee, Won Seok; Park, Jong In; Son, Kwang-Jae; Park, Min; Bang, Young-bong; Choy, Young Bin; Ye, Sung-Joon

    2015-06-01

    In radiotherapy, metallic implants often detach from their deposited sites and migrate to other locations. This undesirable migration could cause inadequate dose coverage for permanent brachytherapy and difficulties in image-guided radiation delivery for patients. To prevent migration of implanted seeds, the authors propose a potential strategy to use a biocompatible and tissue-adhesive material called polydopamine. In this study, nonradioactive dummy seeds that have the same geometry and composition as commercial I-125 seeds were coated in polydopamine. Using scanning electron microscopy and x-ray photoelectron spectroscopy, the surface of the polydopamine-coated and noncoated seeds was characterized. The detachment stress between the two types of seeds and the tissue was measured. The efficacy of polydopamine-coated seed was investigated through in vitro migration tests by tracing the seed location after tissue implantation and shaking for given times. The cytotoxicity of the polydopamine coating was also evaluated. The results of the coating characterization have shown that polydopamine was successfully coated on the surface of the seeds. In the adhesion test, the polydopamine-coated seeds had 2.1-fold greater detachment stress than noncoated seeds. From the in vitro test, it was determined that the polydopamine-coated seed migrated shorter distances than the noncoated seed. This difference was increased with a greater length of time after implantation. The authors suggest that polydopamine coating is an effective technique to prevent migration of implanted seeds, especially for permanent prostate brachytherapy.

  5. Surface coating for prevention of metallic seed migration in tissues

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Hyunseok; Park, Jong In [Program in Biomedical Radiation Sciences, Department of Transdisciplinary Studies, Graduate School of Convergence Science and Technology, Seoul National University, Seoul 151-742 (Korea, Republic of); Lee, Won Seok; Park, Min [Interdisciplinary Program in Bioengineering, Seoul National University College of Engineering, Seoul 151-742 (Korea, Republic of); Son, Kwang-Jae [Hanaro Applications Research, Korea Atomic Energy Research Institute, Daejeon 305-353 (Korea, Republic of); Bang, Young-bong [Advanced Institutes of Convergence Technology, Seoul National University, Suwon 443-270 (Korea, Republic of); Choy, Young Bin, E-mail: ybchoy@snu.ac.kr, E-mail: sye@snu.ac.kr [Interdisciplinary Program in Bioengineering, Seoul National University College of Engineering, Seoul 110-744 (Korea, Republic of); Department of Biomedical Engineering, Seoul National University College of Medicine, Seoul 110-744 (Korea, Republic of); Institute of Medical and Biological Engineering, Medical Research Center, Seoul National University, Seoul 110-744 (Korea, Republic of); Ye, Sung-Joon, E-mail: ybchoy@snu.ac.kr, E-mail: sye@snu.ac.kr [Program in Biomedical Radiation Sciences, Department of Transdisciplinary Studies, Graduate School of Convergence Science and Technology, Seoul National University, Seoul 151-742 (Korea, Republic of); Advanced Institutes of Convergence Technology, Seoul National University, Suwon 443-270 (Korea, Republic of); Department of Radiation Oncology, Seoul National University Hospital, Seoul 110-744 (Korea, Republic of)

    2015-06-15

    Purpose: In radiotherapy, metallic implants often detach from their deposited sites and migrate to other locations. This undesirable migration could cause inadequate dose coverage for permanent brachytherapy and difficulties in image-guided radiation delivery for patients. To prevent migration of implanted seeds, the authors propose a potential strategy to use a biocompatible and tissue-adhesive material called polydopamine. Methods: In this study, nonradioactive dummy seeds that have the same geometry and composition as commercial I-125 seeds were coated in polydopamine. Using scanning electron microscopy and x-ray photoelectron spectroscopy, the surface of the polydopamine-coated and noncoated seeds was characterized. The detachment stress between the two types of seeds and the tissue was measured. The efficacy of polydopamine-coated seed was investigated through in vitro migration tests by tracing the seed location after tissue implantation and shaking for given times. The cytotoxicity of the polydopamine coating was also evaluated. Results: The results of the coating characterization have shown that polydopamine was successfully coated on the surface of the seeds. In the adhesion test, the polydopamine-coated seeds had 2.1-fold greater detachment stress than noncoated seeds. From the in vitro test, it was determined that the polydopamine-coated seed migrated shorter distances than the noncoated seed. This difference was increased with a greater length of time after implantation. Conclusions: The authors suggest that polydopamine coating is an effective technique to prevent migration of implanted seeds, especially for permanent prostate brachytherapy.

  6. Comparison of connective tissue graft and guided tissue regeneration in covering root surfaces

    Directory of Open Access Journals (Sweden)

    LotfazarM.

    2002-08-01

    Full Text Available There are many researches evaluation different methods for covering the root surface. In the most of these studies, type I and II of Miller treatment had been searched. The purpose of this study was a comparison between connective tissue graft (CTG and guided tissue regeneration (GTR with a collagen membrane in the treatment of gingival recession defects (Miller class III. Six patients, each contributing a pair of Miller class III buccal gingival recessions, were treated. The clinical measurements were obtained at baseline and 1,2,4,6,12,18 months after surgery. Statistical analysis were performed using paired t-test between periods (baseline versus 6 months and baseline versus 18 months within each treatment group and also between treatment groups before treatment and 6, 12 and 18 months after the treatment. The treatments were compared by a triple analysis of variance along the time (treatment, patient, time. Both CTG and GTR with a bioabsorbable membrane demonstrated significant clinical and esthetic improvement for gingival recession coverage. The CTG and GTR procedures had mean root coverage of 55% and 47.5% respectively, in the end of study. The CTG group was statistically better than GTR for recession depth, recession width and keratinized tissue width. Also, passing the time (18 months as a distinct factor of treatment procedures was effective in increasing of clinical attachment level and keratinized tissue width.

  7. Effect of the inoculation of a starter culture and vacuum packaging (during resting stage) on the appearance and some microbiological and physicochemical parameters of dry-cured ham.

    Science.gov (United States)

    Sánchez-Molinero, F; Arnau, J

    2008-05-01

    The effect of the inoculation of a starter culture and vacuum packaging (during the resting stage) on dry-cured ham appearance, microbiological and physicochemical parameters was studied. Half of the 36 processed hams were inoculated, after salting, with a commercial starter culture containing lactic-acid bacteria, Gram-positive catalase-positive cocci and yeasts. 18 hams per group (inoculated and non-inoculated) remained vacuum packaged during resting. Microbiological analyses were carried out on the lean surface during processing, on subcutaneous fat tissue at the drying stage and on lean tissue in the finished product. Appearance was evaluated during processing. Physicochemical analyses (NaCl, H(2)O, proteolysis index, a(w)) were done on Semimembranosus and Biceps femoris in the final product. Inoculation caused a reduction of mould growth and oil drip. Vacuum packaging induced increased proteolysis and increases in all microbial counts and a reduction of oil drip, mould growth and weight loss during processing.

  8. Viral RNA levels and env variants in semen and tissues of mature male rhesus macaques infected with SIV by penile inoculation.

    Directory of Open Access Journals (Sweden)

    Francis Fieni

    Full Text Available HIV is shed in semen but the anatomic site of virus entry into the genital secretions is unknown. We determined viral RNA (vRNA levels and the envelope gene sequence in the SIVmac 251 viral populations in the genital tract and semen of 5 adult male rhesus monkeys (Macaca mulatta that were infected after experimental penile SIV infection. Paired blood and semen samples were collected from 1-9 weeks after infection and the monkeys were necropsied eleven weeks after infection. The axillary lymph nodes, testes, epididymis, prostate, and seminal vesicles were collected and vRNA levels and single-genome analysis of the SIVmac251 env variants was performed. At the time of semen collection, blood vRNA levels were between 3.09 and 7.85 log10 vRNA copies/ml plasma. SIV RNA was found in the axillary lymph nodes of all five monkeys and in 3 of 5 monkeys, all tissues examined were vRNA positive. In these 3 monkeys, vRNA levels (log10 SIVgag copies/ug of total tissue RNA in the axillary lymph node (6.48 ± 0.50 were significantly higher than in the genital tract tissues: testis (3.67 ± 2.16; p<0.05, epididymis (3.08 ± 1.19; p<0.0001, prostate (3.36 ± 1.30; p<0.01, and seminal vesicle (2.67 ± 1.50; p<0.0001. Comparison of the SIVmac251 env viral populations in blood plasma, systemic lymph node, and genital tract tissues was performed in two of the macaques. Visual inspection of the Neighbor-Joining phylograms revealed that in both animals, all the sequences were generally distributed evenly among all tissue compartments. Importantly, viral populations in the genital tissues were not distinct from those in the systemic tissues. Our findings demonstrate striking similarity in the viral populations in the blood and male genital tract tissues within 3 months of penile SIV transmission.

  9. Tissue localization, shedding, virus carriage, antibody response, and aerosol transmission of porcine epidemic diarrhea virus (PEDV) following inoculation of 4 week-old feeder pigs

    Science.gov (United States)

    Porcine epidemic diarrhea virus (PEDV) emerged in the U.S. in April 2013 and caused significant losses to the swine industry. The purpose of this investigation was to determine tissue localization, shedding patterns, virus carriage, antibody response, and aerosol transmission of PEDV following inocu...

  10. Accessing inoculation methods of maize and wheat with Azospirillum brasilense.

    Science.gov (United States)

    Fukami, Josiane; Nogueira, Marco Antonio; Araujo, Ricardo Silva; Hungria, Mariangela

    2016-03-01

    The utilization of inoculants containing Azospirillum is becoming more popular due to increasing reports of expressive gains in grain yields. However, incompatibility with pesticides used in seed treatments represents a main limitation for a successful inoculation. Therefore, in this study we searched for alternatives methods for seed inoculation of maize and wheat, aiming to avoid the direct contact of bacteria with pesticides. Different doses of inoculants containing Azospirillum brasilense were employed to perform inoculation in-furrow, via soil spray at sowing and via leaf spray after seedlings had emerged, in comparison to seed inoculation. Experiments were conducted first under greenhouse controlled conditions and then confirmed in the field at different locations in Brazil. In the greenhouse, most parameters measured responded positively to the largest inoculant dose used in foliar sprays, but benefits could also be observed from both in-furrow and soil spray inoculation. However, our results present evidence that field inoculation with plant-growth promoting bacteria must consider inoculant doses, and point to the need of fine adjustments to avoid crossing the threshold of growth stimulation and inhibition. All inoculation techniques increased the abundance of diazotrophic bacteria in plant tissues, and foliar spray improved colonization of leaves, while soil inoculations favored root and rhizosphere colonization. In field experiments, inoculation with A. brasilense allowed for a 25 % reduction in the need for N fertilizers. Our results have identified alternative methods of inoculation that were as effective as the standard seed inoculation that may represent an important strategy to avoid the incompatibility between inoculant bacteria and pesticides employed for seed treatment.

  11. Automatic agar tray inoculation device

    Science.gov (United States)

    Wilkins, J. R.; Mills, S. M.

    1972-01-01

    Automatic agar tray inoculation device is simple in design and foolproof in operation. It employs either conventional inoculating loop or cotton swab for uniform inoculation of agar media, and it allows technician to carry on with other activities while tray is being inoculated.

  12. Development of a greenhouse-based inoculation protocol for the fungus Colletotrichum cereale pathogenic to annual bluegrass (Poa annua

    Directory of Open Access Journals (Sweden)

    Lisa A. Beirn

    2015-08-01

    Full Text Available The fungus Colletotrichum cereale incites anthracnose disease on Poa annua (annual bluegrass turfgrass. Anthracnose disease is geographically widespread throughout the world and highly destructive to cool-season turfgrasses, with infections by C. cereale resulting in extensive turf loss. Comprehensive research aimed at controlling turfgrass anthracnose has been performed in the field, but knowledge of the causal organism and its basic biology is still needed. In particular, the lack of a reliable greenhouse-based inoculation protocol performed under controlled environmental conditions is an obstacle to the study of C. cereale and anthracnose disease. Our objective was to develop a consistent and reproducible inoculation protocol for the two major genetic lineages of C. cereale. By adapting previously successful field-based protocols and combining with components of existing inoculation procedures, the method we developed consistently produced C. cereale infection on two susceptible P. annua biotypes. Approximately 7 to 10 days post-inoculation, plants exhibited chlorosis and thinning consistent with anthracnose disease symptomology. Morphological inspection of inoculated plants revealed visual signs of the fungus (appressoria and acervuli, although acervuli were not always present. After stringent surface sterilization of inoculated host tissue, C. cereale was consistently re-isolated from symptomatic tissue. Real-time PCR detection analysis based on the Apn2 marker confirmed the presence of the pathogen in host tissue, with both lineages of C. cereale detected from all inoculated plants. When a humidifier was not used, no infection developed for any biotypes or fungal isolates tested. The inoculation protocol described here marks significant progress for in planta studies of C. cereale, and will enable scientifically reproducible investigations of the biology, infectivity and lifestyle of this important grass pathogen.

  13. [Design of SCM inoculation device].

    Science.gov (United States)

    Qian, Mingli; Xie, Haiyuan

    2014-01-01

    The first step of bacilli culture is inoculation bacteria on culture medium. Designing a device to increase efficiency of inoculation is significative. The new device is controlled by SCM. The stepper motor can drive the culture medium rotating, accelerating, decelerating, overturn and suspending. The device is high practicability and efficient, let inoculation easy for operator.

  14. RDX in Plant Tissue: Leading to Humification in Surface Soils

    Science.gov (United States)

    2013-01-01

    aromatics in plant tissue may control or alter plant-related transformations and photodegradation. Bio - available carbon from decaying plant tissue may be...TR-13-4 39 Agronomists have shown that high-organic-matter soils reduce the efficacy of the herbicide 2-chloro-4-ethylamino-6-isopropylamino-1,3,5...resulting in the formation of nitroamine and formaldehyde (Hawari et al. 2000). These intermediates can then be further bio -transformed to either

  15. Bone tissue response to plasma-nitrided titanium implant surfaces

    Directory of Open Access Journals (Sweden)

    Emanuela Prado FERRAZ

    2015-02-01

    Full Text Available A current goal of dental implant research is the development of titanium (Ti surfaces to improve osseointegration. Plasma nitriding treatments generate surfaces that favor osteoblast differentiation, a key event to the process of osteogenesis. Based on this, it is possible to hypothesize that plasma-nitrided Ti implants may positively impact osseointegration. Objective The aim of this study was to evaluate the in vivo bone response to Ti surfaces modified by plasma-nitriding treatments. Material and Methods Surface treatments consisted of 20% N2 and 80% H2, 450°C and 1.5 mbar during 1 h for planar and 3 h for hollow cathode. Untreated surface was used as control. Ten implants of each surface were placed into rabbit tibiae and 6 weeks post-implantation they were harvested for histological and histomorphometric analyses. Results Bone formation was observed in contact with all implants without statistically significant differences among the evaluated surfaces in terms of bone-to-implant contact, bone area between threads, and bone area within the mirror area. Conclusion Our results indicate that plasma nitriding treatments generate Ti implants that induce similar bone response to the untreated ones. Thus, as these treatments improve the physico-chemical properties of Ti without affecting its biocompatibility, they could be combined with modifications that favor bone formation in order to develop new implant surfaces.

  16. Bone tissue response to plasma-nitrided titanium implant surfaces.

    Science.gov (United States)

    Ferraz, Emanuela Prado; Sverzut, Alexander Tadeu; Freitas, Gileade Pereira; Sá, Juliana Carvalho; Alves, Clodomiro; Beloti, Marcio Mateus; Rosa, Adalberto Luiz

    2015-01-01

    A current goal of dental implant research is the development of titanium (Ti) surfaces to improve osseointegration. Plasma nitriding treatments generate surfaces that favor osteoblast differentiation, a key event to the process of osteogenesis. Based on this, it is possible to hypothesize that plasma-nitrided Ti implants may positively impact osseointegration. Objective The aim of this study was to evaluate the in vivo bone response to Ti surfaces modified by plasma-nitriding treatments. Material and Methods Surface treatments consisted of 20% N2 and 80% H2, 450°C and 1.5 mbar during 1 h for planar and 3 h for hollow cathode. Untreated surface was used as control. Ten implants of each surface were placed into rabbit tibiae and 6 weeks post-implantation they were harvested for histological and histomorphometric analyses. Results Bone formation was observed in contact with all implants without statistically significant differences among the evaluated surfaces in terms of bone-to-implant contact, bone area between threads, and bone area within the mirror area. Conclusion Our results indicate that plasma nitriding treatments generate Ti implants that induce similar bone response to the untreated ones. Thus, as these treatments improve the physico-chemical properties of Ti without affecting its biocompatibility, they could be combined with modifications that favor bone formation in order to develop new implant surfaces.

  17. Customized Fabrication of Osteochondral Tissue for Articular Joint Surface Repair

    Science.gov (United States)

    2016-09-01

    Scaffolds for Cartilage Tissue Engineering. Frontiers in Bioengineering and Biotechnology . Nothing to Report Other publications, conference papers...between 20% vs. 25% and 30%. Histological analysis of ECM production using Safranin O/fast green and Alcian Blue /fast red staining is shown in...encapsulated constructs visualized by Alcian blue /fast green staining at day 28. (A-C) Staining of PDLLA-PEG 4000 group. Staining decreases as

  18. Customized Fabrication of Osteochondral Tissue for Articular Joint Surface Repair

    Science.gov (United States)

    2015-09-01

    company; or  adoption of new practices. What was the impact on society beyond science and technology ? If there is nothing significant to report during...ORGANIZATION REPORT NUMBER University of Pittsburgh, 450 Technology Drive, Pittsburgh, PA, 15219 9. SPONSORING / MONITORING AGENCY NAME(S) AND...it is a disease of the cartilage, bone and surrounding soft tissue that disables 9-10% of the US population. In the US military, combat and non

  19. Enhanced lubrication on tissue and biomaterial surfaces through peptide-mediated binding of hyaluronic acid.

    Science.gov (United States)

    Singh, Anirudha; Corvelli, Michael; Unterman, Shimon A; Wepasnick, Kevin A; McDonnell, Peter; Elisseeff, Jennifer H

    2014-10-01

    Lubrication is key for the efficient function of devices and tissues with moving surfaces, such as articulating joints, ocular surfaces and the lungs. Indeed, lubrication dysfunction leads to increased friction and degeneration of these systems. Here, we present a polymer-peptide surface coating platform to non-covalently bind hyaluronic acid (HA), a natural lubricant in the body. Tissue surfaces treated with the HA-binding system exhibited higher lubricity values, and in vivo were able to retain HA in the articular joint and to bind ocular tissue surfaces. Biomaterials-mediated strategies that locally bind and concentrate HA could provide physical and biological benefits when used to treat tissue-lubricating dysfunction and to coat medical devices.

  20. Enhanced lubrication on tissue and biomaterial surfaces through peptide-mediated binding of hyaluronic acid

    Science.gov (United States)

    Singh, Anirudha; Corvelli, Michael; Unterman, Shimon A.; Wepasnick, Kevin A.; McDonnell, Peter; Elisseeff, Jennifer H.

    2014-10-01

    Lubrication is key for the efficient function of devices and tissues with moving surfaces, such as articulating joints, ocular surfaces and the lungs. Indeed, lubrication dysfunction leads to increased friction and degeneration of these systems. Here, we present a polymer-peptide surface coating platform to non-covalently bind hyaluronic acid (HA), a natural lubricant in the body. Tissue surfaces treated with the HA-binding system exhibited higher lubricity values, and in vivo were able to retain HA in the articular joint and to bind ocular tissue surfaces. Biomaterials-mediated strategies that locally bind and concentrate HA could provide physical and biological benefits when used to treat tissue-lubricating dysfunction and to coat medical devices.

  1. Efficacy of gaseous chlorine dioxide on inactivation of Escherichia coli O157:H7 on surfaces of artificially inoculated fresh-cut Romaine lettuce leaves

    Science.gov (United States)

    Efficacy of antimicrobial agents in inactivating pathogenic microorganisms on fresh and fresh-cut produce surfaces in limited by the accessibility of such agents to the produce surfaces where the microorganisms are residing. Other factors contribute to this problem include biofilm formation by the m...

  2. Surface-functionalized electrospun nanofibers for tissue engineering and drug delivery.

    Science.gov (United States)

    Yoo, Hyuk Sang; Kim, Taek Gyoung; Park, Tae Gwan

    2009-10-05

    Electrospun nanofibers with a high surface area to volume ratio have received much attention because of their potential applications for biomedical devices, tissue engineering scaffolds, and drug delivery carriers. In order to develop electrospun nanofibers as useful nanobiomaterials, surfaces of electrospun nanofibers have been chemically functionalized for achieving sustained delivery through physical adsorption of diverse bioactive molecules. Surface modification of nanofibers includes plasma treatment, wet chemical method, surface graft polymerization, and co-electrospinning of surface active agents and polymers. A variety of bioactive molecules including anti-cancer drugs, enzymes, cytokines, and polysaccharides were entrapped within the interior or physically immobilized on the surface for controlled drug delivery. Surfaces of electrospun nanofibers were also chemically modified with immobilizing cell specific bioactive ligands to enhance cell adhesion, proliferation, and differentiation by mimicking morphology and biological functions of extracellular matrix. This review summarizes surface modification strategies of electrospun polymeric nanofibers for controlled drug delivery and tissue engineering.

  3. Surface modification of polycaprolactone scaffolds fabricated via selective laser sintering for cartilage tissue engineering.

    Science.gov (United States)

    Chen, Chih-Hao; Lee, Ming-Yih; Shyu, Victor Bong-Hang; Chen, Yi-Chieh; Chen, Chien-Tzung; Chen, Jyh-Ping

    2014-07-01

    Surface modified porous polycaprolactone scaffolds fabricated via rapid prototyping techniques were evaluated for cartilage tissue engineering purposes. Polycaprolactone scaffolds manufactured by selective laser sintering (SLS) were surface modified through immersion coating with either gelatin or collagen. Three groups of scaffolds were created and compared for both mechanical and biological properties. Surface modification with collagen or gelatin improved the hydrophilicity, water uptake and mechanical strength of the pristine scaffold. From microscopic observations and biochemical analysis, collagen-modified scaffold was the best for cartilage tissue engineering in terms of cell proliferation and extracellular matrix production. Chondrocytes/collagen-modified scaffold constructs were implanted subdermally in the dorsal spaces of female nude mice. Histological and immunohistochemical staining of the retrieved implants after 8 weeks revealed enhanced cartilage tissue formation. We conclude that collagen surface modification through immersion coating on SLS-manufactured scaffolds is a feasible scaffold for cartilage tissue engineering in craniofacial reconstruction. Copyright © 2014 Elsevier B.V. All rights reserved.

  4. Surface Modification of Biomaterials in Hard Tissue Applications

    Institute of Scientific and Technical Information of China (English)

    LIU Xuan-yong; DING Chuan-xian; CHU Paul K

    2004-01-01

    Surface modification technologies are quite common in the biomedical field to improve the mechanical,chemical, physical and biological properties of implants such as artificial joint and cardiovascular devices. In this paper, recent progress in the investigation of the bioactivity and biocompatibility enhancement of implants using plasma spraying and plasmabased ion implantation (PIII) is described. Plasma sprayed hydroxyapatite (HA) coatings are commonly used as bioactive coatings but the relatively poor adhesion between the coatings and titanium is one of main disadvantages which have limited their biomedical applications. In our recent studies, novel bioactive coatings, such as wollastonite and dicalcium silicate, were deposited onto titanium to enhance the surfaces bioactivity and biocompatibility. Our results indicate that plasma sprayed wollastonite and dicalcium silicate coatings possess excellent bioactivity as well as relatively high bonding strength. Plasma immersion ion implantation was also employed to improve the anti-corrosion and biological properties of implants.

  5. Conditions of lateral surface confinement that promote tissue-cell integration and inhibit biofilm growth

    NARCIS (Netherlands)

    Wang, Yi; Da Silva Domingues, Joana E.; Subbiandoss, Guruprakash; van der Mei, Henny C.; Busscher, Henk J.; Libera, Matthew

    2014-01-01

    Surfaces with cell adhesiveness modulated at micro length scales can exploit differences between tissue/bacterial cell size, membrane/wall plasticity, and adhesion mechanisms to differentially control tissue-cell/material and bacteria/material interactions. This study explores the short-term interac

  6. Influence of skin surface roughness degree on energy characteristics of light scattered by a biological tissue

    Science.gov (United States)

    Barun, V. V.; Ivanov, A. P.

    2017-05-01

    We present the results of modelling of photometric characteristics of light in soft tissues illuminated by a parallel beam along the normal to the surface, obtained with allowance for the skin roughness parameters and the angular structure of radiation approaching the surface from within the tissue. The depth structure of the fluence rate and the spectra of the diffuse reflection of light by the tissue in the interval of wavelengths 300 - 1000 nm are considered. We discuss the influence of the tilt angle variance of rough surface microelements and light refraction on the studied characteristics. It is shown that these factors lead to the reduction of the radiation flux only in the near-surface tissue layer and practically do not affect the depth of light penetration into the tissue. On the other hand, the degree of the surface roughness and the conditions of its illumination from within the tissue essentially affect the coefficient of diffuse reflection of light and lead to its considerable growth compared to the cases of a smooth interface and completely diffuse illumination, often considered to simplify the theoretical problem solution. The role of the roughness of skin surface is assessed in application to the solution of different direct and inverse problems of biomedical optics.

  7. Experimental inoculation of wild turkeys (Meleagris gallopavo) with Mycobacterium bovis.

    Science.gov (United States)

    Clarke, K R; Fitzgerald, S D; Hattey, J A; Bolin, C A; Berry, D E; Church, S V; Reed, W M

    2006-03-01

    Although avian species are known to be susceptible to infection with Mycobacterium spp. organisms, much remains unknown about the susceptibility of birds to infection with M. bovis. The objective of this current study was to determine if wild turkeys (Meleagris gallopavo) can be infected with M. bovis when inoculated by the oral or intratracheal route. Six turkeys were orally inoculated and another six were inoculated via the trachea with a high dose of M. bovis, 1 x 10(5) CFU/ml. Six turkeys were sham-inoculated controls. Two turkeys from each treatment group were sacrificed on days 30, 60, and 90 postinoculation. There were no gross or microscopic lesions consistent with mycobacteriosis in the 23 inoculated turkeys over the 90-day duration of this study. Fecal cultures were also consistently negative for M. bovis when sampled before inoculation and on days 1, 30, and 60 postinoculation. Two intratracheally inoculated turkeys were positive for M. bovis in visceral tissues at 30 days postinoculation. However, this finding was only indicative of passive persistence of mycobacteria in the tissues and not of infection, as there were no attendant lesions or clinical compromise to support infection. Thus, it can be concluded that young wild turkeys are resistant to infection with M. bovis and, therefore, pose minimal threat as reservoir or spillover hosts for this organism.

  8. Quantitative differences in tissue surface tension influence zebrafish germ layer positioning.

    Science.gov (United States)

    Schötz, Eva-Maria; Burdine, Rebecca D; Jülicher, Frank; Steinberg, Malcolm S; Heisenberg, Carl-Philipp; Foty, Ramsey A

    2008-02-01

    This study provides direct functional evidence that differential adhesion, measurable as quantitative differences in tissue surface tension, influences spatial positioning between zebrafish germ layer tissues. We show that embryonic ectodermal and mesendodermal tissues generated by mRNA-overexpression behave on long-time scales like immiscible fluids. When mixed in hanging drop culture, their cells segregate into discrete phases with ectoderm adopting an internal position relative to the mesendoderm. The position adopted directly correlates with differences in tissue surface tension. We also show that germ layer tissues from untreated embryos, when extirpated and placed in culture, adopt a configuration similar to those of their mRNA-overexpressing counterparts. Down-regulating E-cadherin expression in the ectoderm leads to reduced surface tension and results in phase reversal with E-cadherin-depleted ectoderm cells now adopting an external position relative to the mesendoderm. These results show that in vitro cell sorting of zebrafish mesendoderm and ectoderm tissues is specified by tissue interfacial tensions. We perform a mathematical analysis indicating that tissue interfacial tension between actively motile cells contributes to the spatial organization and dynamics of these zebrafish germ layers in vivo.

  9. Surface modification of polycaprolactone scaffolds fabricated via selective laser sintering for cartilage tissue engineering

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Chih-Hao [Department of Chemical and Materials Engineering, Chang Gung University, Kweishan, Taoyuan 333, Taiwan, ROC (China); Department of Plastic and Reconstructive Surgery, Chang Gung Memorial Hospital, Craniofacial Research Center, Chang Gung University, Kweishann, Taoyuan 333, Taiwan, ROC (China); Lee, Ming-Yih [Graduate Institute of Medical Mechatronics, Chang Gung University, Kweishan, Taoyuan 333, Taiwan, ROC (China); Shyu, Victor Bong-Hang; Chen, Yi-Chieh; Chen, Chien-Tzung [Department of Plastic and Reconstructive Surgery, Chang Gung Memorial Hospital, Craniofacial Research Center, Chang Gung University, Kweishann, Taoyuan 333, Taiwan, ROC (China); Chen, Jyh-Ping, E-mail: jpchen@mail.cgu.edu.tw [Department of Chemical and Materials Engineering, Chang Gung University, Kweishan, Taoyuan 333, Taiwan, ROC (China); Research Center for Industry of Human Ecology, Chang Gung University of Science and Technology, Kweishan, Taoyuan 333, Taiwan, ROC (China)

    2014-07-01

    Surface modified porous polycaprolactone scaffolds fabricated via rapid prototyping techniques were evaluated for cartilage tissue engineering purposes. Polycaprolactone scaffolds manufactured by selective laser sintering (SLS) were surface modified through immersion coating with either gelatin or collagen. Three groups of scaffolds were created and compared for both mechanical and biological properties. Surface modification with collagen or gelatin improved the hydrophilicity, water uptake and mechanical strength of the pristine scaffold. From microscopic observations and biochemical analysis, collagen-modified scaffold was the best for cartilage tissue engineering in terms of cell proliferation and extracellular matrix production. Chondrocytes/collagen-modified scaffold constructs were implanted subdermally in the dorsal spaces of female nude mice. Histological and immunohistochemical staining of the retrieved implants after 8 weeks revealed enhanced cartilage tissue formation. We conclude that collagen surface modification through immersion coating on SLS-manufactured scaffolds is a feasible scaffold for cartilage tissue engineering in craniofacial reconstruction. - Highlights: • Selective laser sintered polycaprolactone scaffolds are prepared. • Scaffolds are surface modified through immersion coating with gelatin or collagen. • Collagen-scaffold is the best for cartilage tissue engineering in vitro. • Chondrocytes/collagen-scaffold reveals enhanced cartilage tissue formation in vivo.

  10. Poorly processed reusable surface disinfection tissue dispensers may be a source of infection.

    Science.gov (United States)

    Kampf, Günter; Degenhardt, Stina; Lackner, Sibylle; Jesse, Katrin; von Baum, Heike; Ostermeyer, Christiane

    2014-01-21

    Reusable surface disinfectant tissue dispensers are used in hospitals in many countries because they allow immediate access to pre-soaked tissues for targeted surface decontamination. On the other hand disinfectant solutions with some active ingredients may get contaminated and cause outbreaks. We determined the frequency of contaminated surface disinfectant solutions in reusable dispensers and the ability of isolates to multiply in different formulations. Reusable tissue dispensers with different surface disinfectants were randomly collected from healthcare facilities. Solutions were investigated for bacterial contamination. The efficacy of two surface disinfectants was determined in suspension tests against two isolated species directly from a contaminated solution or after 5 passages without selection pressure in triplicate. Freshly prepared use solutions were contaminated to determine survival of isolates. 66 dispensers containing disinfectant solutions with surface-active ingredients were collected in 15 healthcare facilities. 28 dispensers from nine healthcare facilities were contaminated with approximately 107 cells per mL of Achromobacter species 3 (9 hospitals), Achromobacter xylosoxidans or Serratia marcescens (1 hospital each). In none of the hospitals dispenser processing had been adequately performed. Isolates regained susceptibility to the disinfectants after five passages without selection pressure but were still able to multiply in different formulations from different manufacturers at room temperature within 7 days. Neglecting adequate processing of surface disinfectant dispensers has contributed to frequent and heavy contamination of use-solutions based on surface active ingredients. Tissue dispenser processing should be taken seriously in clinical practice.

  11. Surface functionalization of nanobiomaterials for application in stem cell culture, tissue engineering, and regenerative medicine.

    Science.gov (United States)

    Rana, Deepti; Ramasamy, Keerthana; Leena, Maria; Jiménez, Constanza; Campos, Javier; Ibarra, Paula; Haidar, Ziyad S; Ramalingam, Murugan

    2016-05-01

    Stem cell-based approaches offer great application potential in tissue engineering and regenerative medicine owing to their ability of sensing the microenvironment and respond accordingly (dynamic behavior). Recently, the combination of nanobiomaterials with stem cells has paved a great way for further exploration. Nanobiomaterials with engineered surfaces could mimic the native microenvironment to which the seeded stem cells could adhere and migrate. Surface functionalized nanobiomaterial-based scaffolds could then be used to regulate or control the cellular functions to culture stem cells and regenerate damaged tissues or organs. Therefore, controlling the interactions between nanobiomaterials and stem cells is a critical factor. However, surface functionalization or modification techniques has provided an alternative approach for tailoring the nanobiomaterials surface in accordance to the physiological surrounding of a living cells; thereby, enhancing the structural and functional properties of the engineered tissues and organs. Currently, there are a variety of methods and technologies available to modify the surface of biomaterials according to the specific cell or tissue properties to be regenerated. This review highlights the trends in surface modification techniques for nanobiomaterials and the biological relevance in stem cell-based tissue engineering and regenerative medicine. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:554-567, 2016.

  12. Development, fabrication and evaluation of a novel biomimetic human breast tissue derived breast implant surface.

    Science.gov (United States)

    Barr, S; Hill, E W; Bayat, A

    2017-02-01

    Breast implant use has tripled in the last decade with over 320,000 breast implant based reconstructions and augmentations performed in the US per annum. Unfortunately a considerable number of women will experience capsular contracture, the irrepressible and disfiguring, tightening and hardening of the fibrous capsule that envelops the implant. Functionalising implant surfaces with biocompatible tissue-specific textures may improve in vivo performance. A novel biomimetic breast implant is presented here with anti-inflammatory in vitro abilities. Topographical assessment of native breast tissue facilitated the development of a statistical model of adipose tissue. 3D grayscale photolithography and ion etching were combined to successfully replicate a surface modelled upon the statistics of breast tissue. Pro-inflammatory genes ILβ1, TNFα, and IL6 were downregulated (p<0.001) and anti-inflammatory gene IL-10 were upregulated on the novel surface. Pro-inflammatory cytokines Gro-Alpha, TNFα and neutrophil chemoattractant IL8 were produced in lower quantities and anti-inflammatory IL-10 in higher quantities in culture with the novel surface (p<0.01). Immunocytochemistry and SEM demonstrated favourable fibroblast and macrophage responses to these novel surfaces. This study describes the first biomimetic breast tissue derived breast implant surface. Our findings attest to its potential translational ability to reduce the inflammatory phase of the implant driven foreign body reaction.

  13. Analytical prediction of sub surface thermal history in translucent tissue phantoms during plasmonic photo thermotherapy

    CERN Document Server

    Dhar, Purbarun; Narasimhan, Arunn; Das, Sarit K

    2015-01-01

    Knowledge of thermal history in biological tissues during laser based hyperthermia is essential to achieve necrosis of tumour orcarcinoma cells. A semi analytical model to predict sub surface thermal history in translucent, soft, bio tissue mimics has been proposed. The model can accurately predict the spatio temporal temperature variations along depth and the anomalous thermal behaviour in such media, viz. occurrence of sub surface temperature peaks. Based on opto thermal properties, the augmented temperature and shift of the peak positions in case of gold nanostructure mediated tissue phantom hyperthermia can be predicted. Employing inverse approach, the absorbance coefficient of nano graphene infused tissue mimics is determined from the peak temperature and found to provide appreciably accurate predictions along depth. Furthermore, a simplistic, dimensionally consistent correlation to theoretically determine the position of the peak in such media is proposed and found to be consistent with experiments and ...

  14. Controlled surface topography regulates collective 3D migration by epithelial-mesenchymal composite embryonic tissues.

    Science.gov (United States)

    Song, Jiho; Shawky, Joseph H; Kim, YongTae; Hazar, Melis; LeDuc, Philip R; Sitti, Metin; Davidson, Lance A

    2015-07-01

    Cells in tissues encounter a range of physical cues as they migrate. Probing single cell and collective migratory responses to physically defined three-dimensional (3D) microenvironments and the factors that modulate those responses are critical to understanding how tissue migration is regulated during development, regeneration, and cancer. One key physical factor that regulates cell migration is topography. Most studies on surface topography and cell mechanics have been carried out with single migratory cells, yet little is known about the spreading and motility response of 3D complex multi-cellular tissues to topographical cues. Here, we examine the response to complex topographical cues of microsurgically isolated tissue explants composed of epithelial and mesenchymal cell layers from naturally 3D organized embryos of the aquatic frog Xenopus laevis. We control topography using fabricated micropost arrays (MPAs) and investigate the collective 3D migration of these multi-cellular systems in these MPAs. We find that the topography regulates both collective and individual cell migration and that dense MPAs reduce but do not eliminate tissue spreading. By modulating cell size through the cell cycle inhibitor Mitomycin C or the spacing of the MPAs we uncover how 3D topographical cues disrupt collective cell migration. We find surface topography can direct both single cell motility and tissue spreading, altering tissue-scale processes that enable efficient conversion of single cell motility into collective movement.

  15. Spatial Quantitation of Drugs in tissues using Liquid Extraction Surface Analysis Mass Spectrometry Imaging

    Science.gov (United States)

    Swales, John G.; Strittmatter, Nicole; Tucker, James W.; Clench, Malcolm R.; Webborn, Peter J. H.; Goodwin, Richard J. A.

    2016-11-01

    Liquid extraction surface analysis mass spectrometry imaging (LESA-MSI) has been shown to be an effective tissue profiling and imaging technique, producing robust and reliable qualitative distribution images of an analyte or analytes in tissue sections. Here, we expand the use of LESA-MSI beyond qualitative analysis to a quantitative analytical technique by employing a mimetic tissue model previously shown to be applicable for MALDI-MSI quantitation. Liver homogenate was used to generate a viable and molecularly relevant control matrix for spiked drug standards which can be frozen, sectioned and subsequently analyzed for the generation of calibration curves to quantify unknown tissue section samples. The effects of extraction solvent composition, tissue thickness and solvent/tissue contact time were explored prior to any quantitative studies in order to optimize the LESA-MSI method across several different chemical entities. The use of a internal standard to normalize regional differences in ionization response across tissue sections was also investigated. Data are presented comparing quantitative results generated by LESA-MSI to LC-MS/MS. Subsequent analysis of adjacent tissue sections using DESI-MSI is also reported.

  16. Spatial Quantitation of Drugs in tissues using Liquid Extraction Surface Analysis Mass Spectrometry Imaging.

    Science.gov (United States)

    Swales, John G; Strittmatter, Nicole; Tucker, James W; Clench, Malcolm R; Webborn, Peter J H; Goodwin, Richard J A

    2016-11-24

    Liquid extraction surface analysis mass spectrometry imaging (LESA-MSI) has been shown to be an effective tissue profiling and imaging technique, producing robust and reliable qualitative distribution images of an analyte or analytes in tissue sections. Here, we expand the use of LESA-MSI beyond qualitative analysis to a quantitative analytical technique by employing a mimetic tissue model previously shown to be applicable for MALDI-MSI quantitation. Liver homogenate was used to generate a viable and molecularly relevant control matrix for spiked drug standards which can be frozen, sectioned and subsequently analyzed for the generation of calibration curves to quantify unknown tissue section samples. The effects of extraction solvent composition, tissue thickness and solvent/tissue contact time were explored prior to any quantitative studies in order to optimize the LESA-MSI method across several different chemical entities. The use of a internal standard to normalize regional differences in ionization response across tissue sections was also investigated. Data are presented comparing quantitative results generated by LESA-MSI to LC-MS/MS. Subsequent analysis of adjacent tissue sections using DESI-MSI is also reported.

  17. Bone Response to Surface-Modified Titanium Implants: Studies on the Early Tissue Response to Implants with Different Surface Characteristics

    Directory of Open Access Journals (Sweden)

    C. Larsson Wexell

    2013-01-01

    Full Text Available In a series of experimental studies, the bone formation around systematically modified titanium implants is analyzed. In the present study, three different surface modifications were prepared and evaluated. Glow-discharge cleaning and oxidizing resulted in a highly stoichiometric TiO2 surface, while a glow-discharge treatment in nitrogen gas resulted in implants with essentially a surface of titanium nitride, covered with a very thin titanium oxide. Finally, hydrogen peroxide treatment of implants resulted in an almost stoichiometric TiO2, rich in hydroxyl groups on the surface. Machined commercially pure titanium implants served as controls. Scanning Auger Electron Spectroscopy, Scanning Electron Microscopy, and Atomic Force Microscopy revealed no significant differences in oxide thickness or surface roughness parameters, but differences in the surface chemical composition and apparent topography were observed. After surface preparation, the implants were inserted in cortical bone of rabbits and evaluated after 1, 3, and 6 weeks. Light microscopic evaluation of the tissue response showed that all implants were in contact with bone and had a large proportion of newly formed bone within the threads after 6 weeks. There were no morphological differences between the four groups. Our study shows that a high degree of bone contact and bone formation can be achieved with titanium implants of different surface composition and topography.

  18. Ultrasound-guided three-dimensional needle steering in biological tissue with curved surfaces

    NARCIS (Netherlands)

    Abayazid, M.; Lopes da Frota Moreira, P.; Shahriari, N.; Patil, S.; Alterovitz, Ron; Misra, S.

    2015-01-01

    In this paper, we present a system capable of automatically steering a bevel-tipped flexible needle under ultrasound guidance toward a physical target while avoiding a physical obstacle embedded in gelatin phantoms and biological tissue with curved surfaces. An ultrasound pre-operative scan is perfo

  19. Experimental evaluation of apparent tissue surface tension based on the exact solution of the Laplace equation

    Science.gov (United States)

    Norotte, C.; Marga, F.; Neagu, A.; Kosztin, I.; Forgacs, G.

    2008-02-01

    The notion of apparent tissue surface tension offered a systematic way to interpret certain morphogenetic processes in early development. It also allowed deducing quantitative information on cellular and molecular parameters that is otherwise difficult to obtain. To accurately determine such tensions we combined novel experiments with the exact solution of the Laplace equation for the profile of a liquid drop under the employed experimental conditions and used the exact solution to evaluate data collected on tissues. Our results confirm that tissues composed of adhesive and motile cells indeed can be characterized in terms of well-defined apparent surface tension. Our experimental technique presents a way to measure liquid interfacial tensions under conditions when known methods fail.

  20. Biomimetic micro∕nanostructured functional surfaces for microfluidic and tissue engineering applications.

    Science.gov (United States)

    Stratakis, E; Ranella, A; Fotakis, C

    2011-03-30

    This paper reviews our work on the application of ultrafast pulsed laser micro∕nanoprocessing for the three-dimensional (3D) biomimetic modification of materials surfaces. It is shown that the artificial surfaces obtained by femtosecond-laser processing of Si in reactive gas atmosphere exhibit roughness at both micro- and nanoscales that mimics the hierarchical morphology of natural surfaces. Along with the spatial control of the topology, defining surface chemistry provides materials exhibiting notable wetting characteristics which are potentially useful for open microfluidic applications. Depending on the functional coating deposited on the laser patterned 3D structures, we can achieve artificial surfaces that are (a) of extremely low surface energy, thus water-repellent and self-cleaned, and (b) responsive, i.e., showing the ability to change their surface energy in response to different external stimuli such as light, electric field, and pH. Moreover, the behavior of different kinds of cells cultured on laser engineered substrates of various wettabilities was investigated. Experiments showed that it is possible to preferentially tune cell adhesion and growth through choosing proper combinations of surface topography and chemistry. It is concluded that the laser textured 3D micro∕nano-Si surfaces with controllability of roughness ratio and surface chemistry can advantageously serve as a novel means to elucidate the 3D cell-scaffold interactions for tissue engineering applications.

  1. The effect of tissue surface modification with collagenase and addition of TGF-beta 3 on the healing potential of meniscal tears repaired with tissue glues in vitro

    NARCIS (Netherlands)

    Bochynska, Agnieszka Izabela; Hannink, Gerjon; Verhoeven, Renate; Grijpma, Dirk W.; Buma, Pieter

    The aim of the current in vitro study was to investigate if tissue surface modification with collagenase and addition of the TGF-beta 3 can increase the number of cells present in meniscus tears repaired with the use of newly developed tissue adhesives based on isocyanate-terminated block

  2. Study of surface carbohydrates in Galba truncatula tissues before and after infection with Fasciola hepatica.

    Science.gov (United States)

    Georgieva, Katya; Georgieva, Liliya; Mizinska-Boevska, Yana; Stoitsova, Stoyanka R

    2016-07-01

    The presence and distribution of surface carbohydrates in the tissues of Galba truncatula snails uninfected or after infection with Fasciola hepatica as well as on the surface of the snail-pathogenic larval stages of the parasite were studied by lectin labelling assay. This is an attempt to find similarities that indicate possible mimicry, utilised by the parasite as an evasion strategy in this snail-trematode system. Different binding patterns were identified on head-foot-mantle, hepatopancreas, genital glands, renopericardial complex of the host as well as of the snail-pathogenic larval stages of F. hepatica. The infection with F. hepatica leads to changes of labelling with Glycine max in the head-mantle cells and Arachis hypogaea in the tubular epithelium of the hepatopancreas. The lectin binding on the other snail tissues is not changed by the development of the larvae. Our data clearly demonstrated the similarity in labelling of G. truncatula tissues and the surface of the snail-pathogenic larval stages of F. hepatica. The role of glycosylation of the contact surfaces of both organisms in relation to the host-parasite interactions is also discussed.

  3. Study of surface carbohydrates in Galba truncatula tissues before and after infection with Fasciola hepatica

    Science.gov (United States)

    Georgieva, Katya; Georgieva, Liliya; Mizinska-Boevska, Yana; Stoitsova, Stoyanka R

    2016-01-01

    The presence and distribution of surface carbohydrates in the tissues of Galba truncatula snails uninfected or after infection with Fasciola hepatica as well as on the surface of the snail-pathogenic larval stages of the parasite were studied by lectin labelling assay. This is an attempt to find similarities that indicate possible mimicry, utilised by the parasite as an evasion strategy in this snail-trematode system. Different binding patterns were identified on head-foot-mantle, hepatopancreas, genital glands, renopericardial complex of the host as well as of the snail-pathogenic larval stages of F. hepatica. The infection with F. hepatica leads to changes of labelling with Glycine max in the head-mantle cells and Arachis hypogaea in the tubular epithelium of the hepatopancreas. The lectin binding on the other snail tissues is not changed by the development of the larvae. Our data clearly demonstrated the similarity in labelling of G. truncatula tissues and the surface of the snail-pathogenic larval stages of F. hepatica. The role of glycosylation of the contact surfaces of both organisms in relation to the host-parasite interactions is also discussed. PMID:27384082

  4. Nanoscale Surface Modifications of Medical Implants for Cartilage Tissue Repair and Regeneration

    Science.gov (United States)

    Griffin, MF; Szarko, M; Seifailan, A; Butler, PE

    2016-01-01

    Background: Natural cartilage regeneration is limited after trauma or degenerative processes. Due to the clinical challenge of reconstruction of articular cartilage, research into developing biomaterials to support cartilage regeneration have evolved. The structural architecture of composition of the cartilage extracellular matrix (ECM) is vital in guiding cell adhesion, migration and formation of cartilage. Current technologies have tried to mimic the cell’s nanoscale microenvironment to improve implants to improve cartilage tissue repair. Methods: This review evaluates nanoscale techniques used to modify the implant surface for cartilage regeneration. Results: The surface of biomaterial is a vital parameter to guide cell adhesion and consequently allow for the formation of ECM and allow for tissue repair. By providing nanosized cues on the surface in the form of a nanotopography or nanosized molecules, allows for better control of cell behaviour and regeneration of cartilage. Chemical, physical and lithography techniques have all been explored for modifying the nanoscale surface of implants to promote chondrocyte adhesion and ECM formation. Conclusion: Future studies are needed to further establish the optimal nanoscale modification of implants for cartilage tissue regeneration. PMID:28217208

  5. Scaffold Free Bio-orthogonal Assembly of 3-Dimensional Cardiac Tissue via Cell Surface Engineering

    Science.gov (United States)

    Rogozhnikov, Dmitry; O’Brien, Paul J.; Elahipanah, Sina; Yousaf , Muhammad N.

    2016-01-01

    There has been tremendous interest in constructing in vitro cardiac tissue for a range of fundamental studies of cardiac development and disease and as a commercial system to evaluate therapeutic drug discovery prioritization and toxicity. Although there has been progress towards studying 2-dimensional cardiac function in vitro, there remain challenging obstacles to generate rapid and efficient scaffold-free 3-dimensional multiple cell type co-culture cardiac tissue models. Herein, we develop a programmed rapid self-assembly strategy to induce specific and stable cell-cell contacts among multiple cell types found in heart tissue to generate 3D tissues through cell-surface engineering based on liposome delivery and fusion to display bio-orthogonal functional groups from cell membranes. We generate, for the first time, a scaffold free and stable self assembled 3 cell line co-culture 3D cardiac tissue model by assembling cardiomyocytes, endothelial cells and cardiac fibroblast cells via a rapid inter-cell click ligation process. We compare and analyze the function of the 3D cardiac tissue chips with 2D co-culture monolayers by assessing cardiac specific markers, electromechanical cell coupling, beating rates and evaluating drug toxicity. PMID:28008983

  6. Scaffold Free Bio-orthogonal Assembly of 3-Dimensional Cardiac Tissue via Cell Surface Engineering

    Science.gov (United States)

    Rogozhnikov, Dmitry; O’Brien, Paul J.; Elahipanah, Sina; Yousaf, Muhammad N.

    2016-12-01

    There has been tremendous interest in constructing in vitro cardiac tissue for a range of fundamental studies of cardiac development and disease and as a commercial system to evaluate therapeutic drug discovery prioritization and toxicity. Although there has been progress towards studying 2-dimensional cardiac function in vitro, there remain challenging obstacles to generate rapid and efficient scaffold-free 3-dimensional multiple cell type co-culture cardiac tissue models. Herein, we develop a programmed rapid self-assembly strategy to induce specific and stable cell-cell contacts among multiple cell types found in heart tissue to generate 3D tissues through cell-surface engineering based on liposome delivery and fusion to display bio-orthogonal functional groups from cell membranes. We generate, for the first time, a scaffold free and stable self assembled 3 cell line co-culture 3D cardiac tissue model by assembling cardiomyocytes, endothelial cells and cardiac fibroblast cells via a rapid inter-cell click ligation process. We compare and analyze the function of the 3D cardiac tissue chips with 2D co-culture monolayers by assessing cardiac specific markers, electromechanical cell coupling, beating rates and evaluating drug toxicity.

  7. Histological examination of non-lactating bovine udders inoculated with Lactobacillus perolens CRL 1724.

    Science.gov (United States)

    Frola, Ignacio D; Pellegrino, Matías S; Magnano, Gabriel; Giraudo, José A; Espeche, María C; Nader-Macias, María E F; Bogni, Cristina I

    2013-02-01

    The effect of intramammary inoculation of Lactobacillus perolens CRL 1724 on bovine udders at drying off was evaluated through histological examination of the canal and cistern tissues. The persistence of the strain in the udder 7 d post inoculation was also determined. Lb. perolens CRL 1724 was recovered from all mammary quarters and no clinical signs or teat damage were observed after inoculation of 10(6) cfu/ml. The udders showed a normal structural aspect and there were no modifications of the milk appearance. Lb. perolens CRL 1724 cells were evidenced on the surface of the epithelial cells of the cistern without causing any morphological modifications or cell alterations. Lb. perolens CRL 1724 produces a mild inflammatory reaction, characterized by recruitment of neutrophils to the epithelial zone and a slight hyperaemia into blood vessels. This preliminary study provides important information for further studies directed towards the inclusion of Lb. perolens CRL 1724 in the design of probiotic products for preventing bovine mastitis in non-lactating dairy cows.

  8. Distinction of gastric cancer tissue based on surface-enhanced Raman spectroscopy

    Science.gov (United States)

    Ma, Jun; Zhou, Hanjing; Gong, Longjing; Liu, Shu; Zhou, Zhenghua; Mao, Weizheng; Zheng, Rong-er

    2012-12-01

    Gastric cancer is one of the most common malignant tumors with high recurrence rate and mortality rate in China. This study aimed to evaluate the diagnostic capability of Surface-enhanced Raman spectroscopy (SERS) based on gold colloids for distinguishing gastric tissues. Gold colloids were directly mixed with the supernatant of homogenized tissues to heighten the Raman signal of various biomolecule. A total of 56 samples were collected from normal (30) and cancer (26). Raman spectra were obtained with a 785nm excitation in the range of 600-1800 cm-1. Significant spectral differences in SERS mainly belong to nucleic acid, proteins and lipids, particularly in the range of 653, 726, 828, 963, 1004, 1032, 1088, 1130, 1243, 1369, 1474, 1596, 1723 cm-1. PCA-LDA algorithms with leave-one-patient-out cross validation yielded diagnostic sensitivities of 90% (27/30), specificities of 88.5% (23/26), and accuracy of 89.3% (50/56), for classification of normal and cancer tissues. The receiver operating characteristic (ROC) surface is 0.917, illustrating the diagnostic utility of SERS together with PCA-LDA to identify gastric cancer from normal tissue. This work demonstrated the SERS techniques can be useful for gastric cancer detection, and it is also a potential technique for accurately identifying cancerous tumor, which is of considerable clinical importance to real-time diagnosis.

  9. Surface position, not signaling from surrounding maternal tissues, specifies aleurone epidermal cell fate in maize.

    Science.gov (United States)

    Gruis, Darren Fred; Guo, Hena; Selinger, David; Tian, Qing; Olsen, Odd-Arne

    2006-07-01

    Maize (Zea mays) endosperm consists of an epidermal-like surface layer of aleurone cells, an underlying body of starchy endosperm cells, and a basal layer of transfer cells. To determine whether surrounding maternal tissues perform a role in specifying endosperm cell fates, a maize endosperm organ culture technique was established whereby the developing endosperm is completely removed from surrounding maternal tissues. Using cell type-specific fluorescence markers, we show that aleurone cell fate specification occurs exclusively in response to surface position and does not require specific, continued maternal signal input. The starchy endosperm and aleurone cell fates are freely interchangeable throughout the lifespan of the endosperm, with internalized aleurone cells converting to starchy endosperm cells and with starchy endosperm cells that become positioned at the surface converting to aleurone cells. In contrast to aleurone and starchy endosperm cells, transfer cells fail to develop in in vitro-grown endosperm, supporting earlier indications that maternal tissue interaction is required to fully differentiate this cell type. Several parameters confirm that the maize endosperm organ cultures described herein retain the main developmental features of in planta endosperm, including fidelity of aleurone mutant phenotypes, temporal and spatial control of cell type-specific fluorescent markers, specificity of cell type transcripts, and control of mitotic cell divisions.

  10. Surface modification and property analysis of biomedical polymers used for tissue engineering.

    Science.gov (United States)

    Ma, Zuwei; Mao, Zhengwei; Gao, Changyou

    2007-11-15

    The response of host organism in macroscopic, cellular and protein levels to biomaterials is, in most cases, closely associated with the materials' surface properties. In tissue engineering, regenerative medicine and many other biomedical fields, surface engineering of the bio-inert synthetic polymers is often required to introduce bioactive species that can promote cell adhesion, proliferation, viability and enhanced ECM-secretion functions. Up to present, a large number of surface engineering techniques for improving biocompatibility have been well established, the work of which generally contains three main steps: (1) surface modification of the polymeric materials; (2) chemical and physical characterizations; and (3) biocompatibility assessment through cell culture. This review focuses on the principles and practices of surface engineering of biomedical polymers with regards to particular aspects depending on the authors' research background and opinions. The review starts with an introduction of principles in designing polymeric biomaterial surfaces, followed by introduction of surface modification techniques to improve hydrophilicity, to introduce reactive functional groups and to immobilize functional protein molecules. The chemical and physical characterizations of the modified biomaterials are then discussed with emphasis on several important issues such as surface functional group density, functional layer thickness, protein surface density and bioactivity. Three most commonly used surface composition characterization techniques, i.e. ATR-FTIR, XPS, SIMS, are compared in terms of their penetration depth. Ellipsometry, CD, EPR, SPR and QCM's principles and applications in analyzing surface proteins are introduced. Finally discussed are frequently applied methods and their principles to evaluate biocompatibility of biomaterials via cell culture. In this section, current techniques and their developments to measure cell adhesion, proliferation, morphology

  11. Transcriptional profiling differences for articular cartilage and repair tissue in equine joint surface lesions

    Directory of Open Access Journals (Sweden)

    Stromberg Arnold J

    2009-09-01

    Full Text Available Abstract Background Full-thickness articular cartilage lesions that reach to the subchondral bone yet are restricted to the chondral compartment usually fill with a fibrocartilage-like repair tissue which is structurally and biomechanically compromised relative to normal articular cartilage. The objective of this study was to evaluate transcriptional differences between chondrocytes of normal articular cartilage and repair tissue cells four months post-microfracture. Methods Bilateral one-cm2 full-thickness defects were made in the articular surface of both distal femurs of four adult horses followed by subchondral microfracture. Four months postoperatively, repair tissue from the lesion site and grossly normal articular cartilage from within the same femorotibial joint were collected. Total RNA was isolated from the tissue samples, linearly amplified, and applied to a 9,413-probe set equine-specific cDNA microarray. Eight paired comparisons matched by limb and horse were made with a dye-swap experimental design with validation by histological analyses and quantitative real-time polymerase chain reaction (RT-qPCR. Results Statistical analyses revealed 3,327 (35.3% differentially expressed probe sets. Expression of biomarkers typically associated with normal articular cartilage and fibrocartilage repair tissue corroborate earlier studies. Other changes in gene expression previously unassociated with cartilage repair were also revealed and validated by RT-qPCR. Conclusion The magnitude of divergence in transcriptional profiles between normal chondrocytes and the cells that populate repair tissue reveal substantial functional differences between these two cell populations. At the four-month postoperative time point, the relative deficiency within repair tissue of gene transcripts which typically define articular cartilage indicate that while cells occupying the lesion might be of mesenchymal origin, they have not recapitulated differentiation to

  12. Biodegradable polyester-based microcarriers with modified surface tailored for tissue engineering.

    Science.gov (United States)

    Privalova, A; Markvicheva, E; Sevrin, Ch; Drozdova, M; Kottgen, C; Gilbert, B; Ortiz, M; Grandfils, Ch

    2015-03-01

    Microcarriers have been proposed in tissue engineering, namely for bone, cartilage, skin, vascular, and central nervous system. Although polyester-based microcarriers have been already used for this purpose, their surface properties should be improved to provide better cell growth. The goal of this study was to prepare microbeads based on poly(D,L-lactide) acid, poly(L-lactide) acid, and to study cell behavior (adhesion, spreading, growth, and proliferation) in function of microbead topography and surface chemistry. To improve L-929 fibroblasts adhesion, microbead surface has been modified with three polycations: chitosan, poly(2-dimethylamino ethylmethacrylate) (PDMAEMA), or chitosan-g-oligolactide copolymer (chit-g-OLA). Although modification of the microbead surface with chitosan and PDMAEMA was performed through physical adsorption on the previously prepared microbeads, chit-g-OLA copolymer was introduced directly during microbead processing. This simple approach (1) bypass the use of an emulsifier (polyvinyl alcohol, PVA); (2) avoid surface "contamination" with PVA molecules limiting a control of the surface characteristics. In vitro study of the growth of mouse fibroblasts on the microbeads showed that both surface topography and chemistry affected cell attachment, spreading, and proliferation. Cultivation of L-929 fibroblasts for 7 days resulted in the formation of a 3D cell-scaffold network.

  13. Effective reprocessing of reusable dispensers for surface disinfection tissues - the devil is in the details.

    Science.gov (United States)

    Kampf, Günter; Degenhardt, Stina; Lackner, Sibylle; Ostermeyer, Christiane

    2014-01-01

    It has recently been reported that reusable dispensers for surface disinfection tissues may be contaminated, especially with adapted Achromobacter species 3, when products based on surface-active ingredients are used. Fresh solution may quickly become recontaminated if dispensers are not processed adequately. We evaluated the abilities of six manual and three automatic processes for processing contaminated dispensers to prevent recolonisation of a freshly-prepared disinfectant solution (Mikrobac forte 0.5%). Dispensers were left at room temperature for 28 days. Samples of the disinfectant solution were taken every 7 days and assessed quantitatively for bacterial contamination. All automatic procedures prevented recolonisation of the disinfectant solution when a temperature of 60-70°C was ensured for at least 5 min, with or without the addition of chemical cleaning agents. Manual procedures prevented recontamination of the disinfectant solution when rinsing with hot water or a thorough cleaning step was performed before treating all surfaces with an alcohol-based disinfectant or an oxygen-releaser. Other cleaning and disinfection procedures, including the use of an alcohol-based disinfectant, did not prevent recolonisation. These results indicate that not all processes are effective for processing reusable dispensers for surface-disinfectant tissues, and that a high temperature during the cleaning step or use of a biofilm-active cleaning agent are essential.

  14. Effective reprocessing of reusable dispensers for surface disinfection tissues – the devil is in the details

    Science.gov (United States)

    Kampf, Günter; Degenhardt, Stina; Lackner, Sibylle; Ostermeyer, Christiane

    2014-01-01

    Background: It has recently been reported that reusable dispensers for surface disinfection tissues may be contaminated, especially with adapted Achromobacter species 3, when products based on surface-active ingredients are used. Fresh solution may quickly become recontaminated if dispensers are not processed adequately. Methods: We evaluated the abilities of six manual and three automatic processes for processing contaminated dispensers to prevent recolonisation of a freshly-prepared disinfectant solution (Mikrobac forte 0.5%). Dispensers were left at room temperature for 28 days. Samples of the disinfectant solution were taken every 7 days and assessed quantitatively for bacterial contamination. Results: All automatic procedures prevented recolonisation of the disinfectant solution when a temperature of 60–70°C was ensured for at least 5 min, with or without the addition of chemical cleaning agents. Manual procedures prevented recontamination of the disinfectant solution when rinsing with hot water or a thorough cleaning step was performed before treating all surfaces with an alcohol-based disinfectant or an oxygen-releaser. Other cleaning and disinfection procedures, including the use of an alcohol-based disinfectant, did not prevent recolonisation. Conclusions: These results indicate that not all processes are effective for processing reusable dispensers for surface-disinfectant tissues, and that a high temperature during the cleaning step or use of a biofilm-active cleaning agent are essential. PMID:24653973

  15. Precise manipulation of cell behaviors on surfaces for construction of tissue/organs.

    Science.gov (United States)

    Zheng, Wenfu; Jiang, Xingyu

    2014-12-01

    The use of micro/nanotechnology has become an indispensable strategy to manipulating cell microenvironments. By employing key elements of soft lithographical technologies including self-assembled monolayers (SAMs), microcontact printing (μCP), and microfluidic pattering (μFP) and a number of switchable surfaces such as electrochemical active, photosensitive, and thermosensitive surfaces, scientists can control the adhesion, proliferation, migration and differentiation of cells. By combining essential in vivo conditions, various physical or pathological processes such as cell-cell interaction in wound healing and tumor metastasis could be studied on well-defined surfaces and interfaces. By integrating key elements in live tissues, in vitro models mimicking basic structure and function of vital organs such as lung, heart, blood vessel, liver, kidney, and brain have been developed and greatly increased our knowledge of these important life processes. In this review, we will focus on the recent development of these interfacial methods and their application in fundamental biology research.

  16. Viability of multi-strain mixtures of Listeria monocytogenes, Salmonella typhimurium, or Escherichia coli O157:H7 inoculated into the batter or onto the surface of a soudjouk-style fermented semi-dry sausage.

    Science.gov (United States)

    Porto-Fett, A C S; Hwang, C-A; Call, J E; Juneja, V K; Ingham, S C; Ingham, B H; Luchansky, J B

    2008-09-01

    The fate of Listeria monocytogenes, Salmonella typhimurium, or Escherichia coli O157:H7 were separately monitored both in and on soudjouk. Fermentation and drying alone reduced numbers of L. monocytogenes by 0.07 and 0.74 log(10)CFU/g for sausages fermented to pH 5.3 and 4.8, respectively, whereas numbers of S. typhimurium and E. coli O157:H7 were reduced by 1.52 and 3.51 log(10)CFU/g and 0.03 and 1.11 log(10)CFU/g, respectively. When sausages fermented to pH 5.3 or 4.8 were stored at 4, 10, or 21 degrees C, numbers of L. monocytogenes, S. typhimurium, and E. coli O157:H7 decreased by an additional 0.08-1.80, 0.88-3.74, and 0.68-3.17 log(10)CFU/g, respectively, within 30 days. Storage for 90 days of commercially manufactured soudjouk that was sliced and then surface inoculated with L. monocytogenes, S. typhimurium, and E. coli O157:H7 generated average D-values of ca. 10.1, 7.6, and 5.9 days at 4 degrees C; 6.4, 4.3, and 2.9 days at 10 degrees C; 1.4, 0.9, and 1.6 days at 21 degrees C; and 0.9, 1.4, and 0.25 days at 30 degrees C. Overall, fermentation to pH 4.8 and storage at 21 degrees C was the most effective treatment for reducing numbers of L. monocytogenes (2.54 log(10)CFU/g reduction), S. typhimurium (> or =5.23 log(10)CFU/g reduction), and E. coli O157:H7 (3.48 log(10)CFU/g reduction). In summary, soudjouk-style sausage does not provide a favorable environment for outgrowth/survival of these three pathogens.

  17. [Investigation of the surface layer of 3D-matrices for tissue engineering].

    Science.gov (United States)

    Chernonosova, V S; Kvon, R I; Kiseleva, E V; Stepanova, A O; Laktionov, P P

    2017-01-01

    Electrospinning is a convenient and promising manufacturing method a variety of materials for tissue engineering. 3D matrices fabricated by electrospinning from solutions of polycaprolactone with human serum albumin or gelatin in 1,1,1,3,3,3-hexafluoroisopropanol were studied. The microstructure of the 3D matrices and surface of the fibers were investigated using scanning electron microscopy. Protein distribution in the surface layer was studied by modification of protein amino groups with N-(2-hydroxyethyl)phenazine and X-ray photoelectron spectroscopy. It was shown, that concentration of the proteins in the surface layer of fibers exceeded their concentration in the initial electrospun solution up to 12 times and the surface layer was enriched in the protein inversely to the concentration of the protein in solution. The minor part of the proteins was released from fibers during first 30-60 min after swelling in water. Treatment of matrices with proteinase K hydrolyzed about 1/3 of the surface exposed human serum albumin. Thus, both methods can be used to study the surface content of the materials produced by electrospinning from blends of synthetic and natural polymers, however X-ray photoelectron spectroscopy appears to be more convenient and informative.

  18. Surface biofunctionalization of β-TCP blocks using aptamer 74 for bone tissue engineering.

    Science.gov (United States)

    Ardjomandi, N; Huth, J; Stamov, D R; Henrich, A; Klein, C; Wendel, H-P; Reinert, S; Alexander, D

    2016-10-01

    Successful bone regeneration following oral and maxillofacial surgeries depends on efficient functionalization strategies that allow the recruitment of osteogenic progenitor cells at the tissue/implant interface. We have previously identified aptamer 74, which exhibited a binding affinity for osteogenically induced jaw periosteal cells (JPCs). In the present study, this aptamer was used for the surface biofunctionalization of β-tricalcium phosphate (β-TCP) blocks. Atomic force microscopy (AFM) measurements showed increased binding activity of aptamer 74 towards osteogenically induced JPCs compared to untreated controls. The immobilization efficiency of aptamer 74 was analyzed using the QuantiFluor ssDNA assay for 2D surfaces and by amino acid analysis for 3D β-TCP constructs. Following the successful immobilization of aptamer 74 in 2D culture wells and on 3D constructs, in vitro assays showed no significant differences in cell proliferation compared to unmodified surfaces. Interestingly, JPC mineralization was significantly higher on the 2D surfaces and higher cell adhesion was detected on the 3D constructs with immobilized aptamer. Herein, we report an established, biocompatible β-TCP matrix with surface immobilization of aptamer 74, which enhances properties such as cell adhesion on 3D constructs and mineralization on 2D surfaces. Further studies need to be performed to improve the immobilization efficiency and to develop a suitable approach for JPC mineralization growing within 3D β-TCP constructs. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. Controlled implant/soft tissue interaction by nanoscale surface modifications of 3D porous titanium implants

    Science.gov (United States)

    Rieger, Elisabeth; Dupret-Bories, Agnès; Salou, Laetitia; Metz-Boutigue, Marie-Helene; Layrolle, Pierre; Debry, Christian; Lavalle, Philippe; Engin Vrana, Nihal

    2015-05-01

    Porous titanium implants are widely employed in the orthopaedics field to ensure good bone fixation. Recently, the use of porous titanium implants has also been investigated in artificial larynx development in a clinical setting. Such uses necessitate a better understanding of the interaction of soft tissues with porous titanium structures. Moreover, surface treatments of titanium have been generally evaluated in planar structures, while the porous titanium implants have complex 3 dimensional (3D) architectures. In this study, the determining factors for soft tissue integration of 3D porous titanium implants were investigated as a function of surface treatments via quantification of the interaction of serum proteins and cells with single titanium microbeads (300-500 μm in diameter). Samples were either acid etched or nanostructured by anodization. When the samples are used in 3D configuration (porous titanium discs of 2 mm thickness) in vivo (in subcutis of rats for 2 weeks), a better integration was observed for both anodized and acid etched samples compared to the non-treated implants. If the implants were also pre-treated with rat serum before implantation, the integration was further facilitated. In order to understand the underlying reasons for this effect, human fibroblast cell culture tests under several conditions (directly on beads, beads in suspension, beads encapsulated in gelatin hydrogels) were conducted to mimic the different interactions of cells with Ti implants in vivo. Physical characterization showed that surface treatments increased hydrophilicity, protein adsorption and roughness. Surface treatments also resulted in improved adsorption of serum albumin which in turn facilitated the adsorption of other proteins such as apolipoprotein as quantified by protein sequencing. The cellular response to the beads showed considerable difference with respect to the cell culture configuration. When the titanium microbeads were entrapped in cell

  20. Slide-free histology via MUSE: UV surface excitation microscopy for imaging unsectioned tissue (Conference Presentation)

    Science.gov (United States)

    Levenson, Richard M.; Harmany, Zachary; Demos, Stavros G.; Fereidouni, Farzad

    2016-03-01

    Widely used methods for preparing and viewing tissue specimens at microscopic resolution have not changed for over a century. They provide high-quality images but can involve time-frames of hours or even weeks, depending on logistics. There is increasing interest in slide-free methods for rapid tissue analysis that can both decrease turn-around times and reduce costs. One new approach is MUSE (microscopy with UV surface excitation), which exploits the shallow penetration of UV light to excite fluorescent signals from only the most superficial tissue elements. The method is non-destructive, and eliminates requirement for conventional histology processing, formalin fixation, paraffin embedding, or thin sectioning. It requires no lasers, confocal, multiphoton or optical coherence tomography optics. MUSE generates diagnostic-quality histological images that can be rendered to resemble conventional hematoxylin- and eosin-stained samples, with enhanced topographical information, from fresh or fixed, but unsectioned tissue, rapidly, with high resolution, simply and inexpensively. We anticipate that there could be widespread adoption in research facilities, hospital-based and stand-alone clinical settings, in local or regional pathology labs, as well as in low-resource environments.

  1. Inoculation effects on root-colonizing arbuscular mycorrhizal fungal communities spread beyond directly inoculated plants.

    Science.gov (United States)

    Janoušková, Martina; Krak, Karol; Vosátka, Miroslav; Püschel, David; Štorchová, Helena

    2017-01-01

    Inoculation with arbuscular mycorrhizal fungi (AMF) may improve plant performance at disturbed sites, but inoculation may also suppress root colonization by native AMF and decrease the diversity of the root-colonizing AMF community. This has been shown for the roots of directly inoculated plants, but little is known about the stability of inoculation effects, and to which degree the inoculant and the inoculation-induced changes in AMF community composition spread into newly emerging seedlings that were not in direct contact with the introduced propagules. We addressed this topic in a greenhouse experiment based on the soil and native AMF community of a post-mining site. Plants were cultivated in compartmented pots with substrate containing the native AMF community, where AMF extraradical mycelium radiating from directly inoculated plants was allowed to inoculate neighboring plants. The abundances of the inoculated isolate and of native AMF taxa were monitored in the roots of the directly inoculated plants and the neighboring plants by quantitative real-time PCR. As expected, inoculation suppressed root colonization of the directly inoculated plants by other AMF taxa of the native AMF community and also by native genotypes of the same species as used for inoculation. In the neighboring plants, high abundance of the inoculant and the suppression of native AMF were maintained. Thus, we demonstrate that inoculation effects on native AMF propagate into plants that were not in direct contact with the introduced inoculum, and are therefore likely to persist at the site of inoculation.

  2. A Surface Heat Disturbance Method for Measuring Local TIssue Blood Perfusion Rate

    Institute of Scientific and Technical Information of China (English)

    PengJianshu; TianYongquan

    1996-01-01

    A non-damage method for measuring local tissue blood perfusion rate by surface heat disturbance and its two special embodiments for realizing this measurement are presented in this paper,In the derivation of mathematical model,the Pennes equation is used,and two parameters which are very diffcult to be known-the arterial blood temperature Ta and the metabolic heat generation rate qm,have been eliminated.So if has provided a feasible basis for application.In this paper,the performance and the adaptable condition of the measurement method and its structure peculiarities are discussed over a wide variety of parameters.Moreover,the effectiveness of the measurement method has been demonstrated by means of the quantitative measurement of tissue in vitro,the comparative measurement of animal under artificial perfusion and the dependence measurement in human body.

  3. Backscattering of linearly polarized light from turbid tissue-like scattering medium with rough surface

    Science.gov (United States)

    Doronin, Alexander; Tchvialeva, Lioudmila; Markhvida, Igor; Lee, Tim K.; Meglinski, Igor

    2016-07-01

    In the framework of further development of a unified computational tool for the needs of biomedical optics, we introduce an electric field Monte Carlo (MC) model for simulation of backscattering of coherent linearly polarized light from a turbid tissue-like scattering medium with a rough surface. We consider the laser speckle patterns formation and the role of surface roughness in the depolarization of linearly polarized light backscattered from the medium. The mutual phase shifts due to the photons' pathlength difference within the medium and due to reflection/refraction on the rough surface of the medium are taken into account. The validation of the model includes the creation of the phantoms of various roughness and optical properties, measurements of co- and cross-polarized components of the backscattered/reflected light, its analysis and extensive computer modeling accelerated by parallel computing on the NVIDIA graphics processing units using compute unified device architecture (CUDA). The analysis of the spatial intensity distribution is based on second-order statistics that shows a strong correlation with the surface roughness, both with the results of modeling and experiment. The results of modeling show a good agreement with the results of experimental measurements on phantoms mimicking human skin. The developed MC approach can be used for the direct simulation of light scattered by the turbid scattering medium with various roughness of the surface.

  4. Can surface preparation with CVD diamond tip influence on bonding to dental tissues?

    Science.gov (United States)

    Aparecido Kawaguchi, Fernando; Brossi Botta, Sergio; Nilo Vieira, Samuel; Steagall Júnior, Washington; Bona Matos, Adriana

    2008-04-01

    This study evaluated the influence of chemical vapor deposition (CVD) tips surface treatments of enamel and dentin on bonding resistance of two adhesive systems. Thirty embedded samples were divided in 12 groups ( n = 10), according to factors: substrate (enamel and dentin), adhesive system [etch-and-rinse (SB) and self-etch]; and the surface treatments (paper discs, impact CVD tips and tangential CVD tip). When CVD tip was used in the impact mode the tip was applied perpendicular to dental surface, while at tangential mode, the tip worked parallel to dental surface. Specimens were tested in tension after 24 h at 0.5 mm/min of cross-head speed. ANOVA results, in MPa showed that in enamel, only adhesive system factor was statistically significant ( p = 0.015) under tested conditions, with higher bond strength observed for SB groups. However, in dentin the best bonding performance was obtained in SE groups ( p = 0.00). In both tested substrates, results did not show statistically significant difference for factors treatment and its interactions. ConclusionsIt may be concluded that CVD-tip surface treatment, in both tested modes, did not influence on adhesion to enamel and dentin. But, it is important to choose adhesive system according to the tissue available to bonding.

  5. Chondroitin sulfate immobilization at the surface of electrospun nanofiber meshes for cartilage tissue regeneration approaches

    Science.gov (United States)

    Piai, Juliana Francis; da Silva, Marta Alves; Martins, Albino; Torres, Ana Bela; Faria, Susana; Reis, Rui L.; Muniz, Edvani Curti; Neves, Nuno M.

    2017-05-01

    Aiming at improving the biocompatibility of biomaterial scaffolds, surface modification presents a way to preserve their mechanical properties and to improve the surface bioactivity. In this work, chondroitin sulfate (CS) was immobilized at the surface of electrospun poly(caprolactone) nanofiber meshes (PCL NFMs), previously functionalized by UV/O3 exposure and aminolysis. Contact angle, SEM, optical profilometry, FTIR, X-ray photoelectron spectroscopy techniques confirmed the success of CS-immobilization in PCL NFMs. Furthermore, CS-immobilized PCL NFMs showed lower roughness and higher hydrophilicity than the samples without CS. Human articular chondrocytes (hACs) were cultured on electrospun PCL NFMs with or without CS immobilization. It was observed that hACs proliferated through the entire time course of the experiment in both types of nanofibrous scaffolds, as well as for the production of glycosaminoglycans. Quantitative-PCR results demonstrated over-expression of cartilage-related genes such as Aggrecan, Collagen type II, COMP and Sox9 on both types of nanofibrous scaffolds. Morphological observations from SEM and LSCM revealed that hACs maintained their characteristic round shape and cellular agglomeration exclusively on PCL NFMs with CS immobilization. In conclusion, CS immobilization at the surface of PCL NFMs was achieved successfully and provides a valid platform enabling further surface functionalization methods in scaffolds to be developed for cartilage tissue engineering.

  6. A dynamic cell adhesion surface regulates tissue architecture in growth plate cartilage.

    Science.gov (United States)

    Romereim, Sarah M; Conoan, Nicholas H; Chen, Baojiang; Dudley, Andrew T

    2014-05-01

    The architecture and morphogenetic properties of tissues are founded in the tissue-specific regulation of cell behaviors. In endochondral bones, the growth plate cartilage promotes bone elongation via regulated chondrocyte maturation within an ordered, three-dimensional cell array. A key event in the process that generates this cell array is the transformation of disordered resting chondrocytes into clonal columns of discoid proliferative cells aligned with the primary growth vector. Previous analysis showed that column-forming chondrocytes display planar cell divisions, and the resulting daughter cells rearrange by ∼90° to align with the lengthening column. However, these previous studies provided limited information about the mechanisms underlying this dynamic process. Here we present new mechanistic insights generated by application of a novel time-lapse confocal microscopy method along with immunofluorescence and electron microscopy. We show that, during cell division, daughter chondrocytes establish a cell-cell adhesion surface enriched in cadherins and β-catenin. Rearrangement into columns occurs concomitant with expansion of this adhesion surface in a process more similar to cell spreading than to migration. Column formation requires cell-cell adhesion, as reducing cadherin binding via chelation of extracellular calcium inhibits chondrocyte rearrangement. Importantly, physical indicators of cell polarity, such as cell body alignment, are not prerequisites for oriented cell behavior. Our results support a model in which regulation of adhesive surface dynamics and cortical tension by extrinsic signaling modifies the thermodynamic landscape to promote organization of daughter cells in the context of the three-dimensional growth plate tissue.

  7. Priming the Surface of Orthopedic Implants for Osteoblast Attachment in Bone Tissue Engineering.

    Science.gov (United States)

    Chan, Kiat Hwa; Zhuo, Shuangmu; Ni, Ming

    2015-01-01

    The development of better orthopedic implants is incessant. While current implants can function reliably in the human body for a long period of time, there are still a significant number of cases for which the implants can fail prematurely due to poor osseointegration of the implant with native bone. Increasingly, it is recognized that it is extremely important to facilitate the attachment of osteoblasts on the implant so that a proper foundation of extracellular matrix (ECM) can be laid down for the growth of new bone tissue. In order to facilitate the osseointegration of the implant, both the physical nanotopography and chemical functionalization of the implant surface have to be optimized. In this short review, however, we explore how simple chemistry procedures can be used to functionalize the surfaces of three major classes of orthopedic implants, i.e. ceramics, metals, and polymers, so that the attachment of osteoblasts on implants can be facilitated in order to promote implant osseointegration.

  8. Assessment of the Low Alloy Cast Steel Inoculation Effects with Chosen Additives

    Directory of Open Access Journals (Sweden)

    D. Bartocha

    2012-04-01

    Full Text Available Structure, and thus the mechanical properties of steel are primarily a function of chemical composition and the solidification process which can be influenced by the application of the inoculation treatment. This effect depends on the modifier used. The article presents the results of studies designed to assess the effects of structural low alloy steel inoculation by selected modifying additives. The study was performed on nine casts modeled with different inoculants, assessment of the procedure impact was based on the macrostructure of made castings. The ratio of surface area equivalent to the axial zone of the crystals and columnar crystals zone was adopted as a measure of the inoculation effect.

  9. Experimental inoculation of juvenile rhesus macaques with primate enteric caliciviruses.

    Directory of Open Access Journals (Sweden)

    Karol Sestak

    Full Text Available BACKGROUND: Tissue culture-adapted Tulane virus (TV, a GI.1 rhesus enteric calicivirus (ReCV, and a mixture of GII.2 and GII.4 human norovirus (NoV-containing stool sample were used to intrastomacheally inoculate juvenile rhesus macaques (Macaca mulatta in order to evaluate infection caused by these viruses. METHODOLOGY & FINDINGS: Two of the three TV-inoculated macaques developed diarrhea, fever, virus-shedding in stools, inflammation of duodenum and 16-fold increase of TV-neutralizing (VN serum antibodies but no vomiting or viremia. No VN-antibody responses could be detected against a GI.2 ReCV strain FT285, suggesting that TV and FT285 represent different ReCV serotypes. Both NoV-inoculated macaques remained asymptomatic but with demonstrable virus shedding in one animal. Examination of duodenum biopsies of the TV-inoculated macaques showed lymphocytic infiltration of the lamina propria and villous blunting. TV antigen-positive (TV+ cells were detected in the lamina propria. In most of the TV+ cells TV co-localized perinuclearly with calnexin--an endoplasmic reticulum protein. A few CD20+TV+ double-positive B cells were also identified in duodenum. To corroborate the authenticity of CD20+TV+ B cells, in vitro cultures of peripheral blood mononuclear cells (PBMCs from healthy macaques were inoculated with TV. Multicolor flow cytometry confirmed the presence of TV antigen-containing B cells of predominantly CD20+HLA-DR+ phenotype. A 2-log increase of viral RNA by 6 days post inoculation (p<0.05 suggested active TV replication in cultured lymphocytes. CONCLUSIONS/SIGNIFICANCE: Taken together, our results show that ReCVs represent an alternative cell culture and animal model to study enteric calicivirus replication, pathogenesis and immunity.

  10. Platelets interact with tissue factor immobilized on surfaces: effects of shear rate.

    Science.gov (United States)

    Tonda, R; Lopez-Vilchez, I; Navalon, F; Pino, M; Hernandez, M R; Escolar, G; Galan, A M

    2008-01-01

    While procoagulant activities of Tissue Factor (TF) have been widely investigated, its possible pro-adhesive properties towards platelets have not been studied in detail. We explored the interaction of platelets with human Tissue Factor (hTF) firmly adsorbed on a synthetic surface of polyvinilidene difluoride (PVDF) using different shear rates. For studies at 250 and 600 s(-1), TF firmly adsorbed was exposed to flowing anticoagulated blood in flat perfusion devices. Deposition of platelets and fibrin were evaluated by morphometric, immunocytochemical and ultrastructural methods. Prothrombin fragment 1 + 2 (F1 + 2) levels were also measured. Experiments at 5000 s(-1), were performed on the Platelet Function Analyzer (PFA-100) with experimental cartridges with collagen (COL) or collagen-hTF (COL + TF). Haemostatic effect of recombinant activated FVIIa (rFVIIa) was assessed in the same experimental settings. Platelet deposition on hTF reached 19.8 +/- 1.3% and 26.1 +/- 3.4% of the total surface, at 250 and 600 s(-1), respectively. Fibrin formation was significantly higher at 250 s(-1) than at 600 s(-1) (P hTF (154.09 +/- 14.69 s vs. 191.45 +/- 16.09 s COL alone; P hTF is an adhesive substrate for platelets and suggest that the von Willebrand factor could mediate these interactions. At low and intermediate shear rates, rFVIIa enhanced the procoagulant action of hTF, but this effect was not observed at very high shear rates.

  11. Fine-tuning of substrate architecture and surface chemistry promotes muscle tissue development.

    Science.gov (United States)

    Guex, A G; Kocher, F M; Fortunato, G; Körner, E; Hegemann, D; Carrel, T P; Tevaearai, H T; Giraud, M N

    2012-04-01

    Tissue engineering has been increasingly brought to the scientific spotlight in response to the tremendous demand for regeneration, restoration or substitution of skeletal or cardiac muscle after traumatic injury, tumour ablation or myocardial infarction. In vitro generation of a highly organized and contractile muscle tissue, however, crucially depends on an appropriate design of the cell culture substrate. The present work evaluated the impact of substrate properties, in particular morphology, chemical surface composition and mechanical properties, on muscle cell fate. To this end, aligned and randomly oriented micron (3.3±0.8 μm) or nano (237±98 nm) scaled fibrous poly(ε-caprolactone) non-wovens were processed by electrospinning. A nanometer-thick oxygen functional hydrocarbon coating was deposited by a radio frequency plasma process. C2C12 muscle cells were grown on pure and as-functionalized substrates and analysed for viability, proliferation, spatial orientation, differentiation and contractility. Cell orientation has been shown to depend strongly on substrate architecture, being most pronounced on micron-scaled parallel-oriented fibres. Oxygen functional hydrocarbons, representing stable, non-immunogenic surface groups, were identified as strong triggers for myotube differentiation. Accordingly, the highest myotube density (28±15% of total substrate area), sarcomeric striation and contractility were found on plasma-coated substrates. The current study highlights the manifold material characteristics to be addressed during the substrate design process and provides insight into processes to improve bio-interfaces.

  12. A novel surface modification on calcium polyphosphate scaffold for articular cartilage tissue engineering

    Energy Technology Data Exchange (ETDEWEB)

    Lien, S.-M. [Department of Chemical Engineering, National Tsing Hua University, Hsinchu 300 (China); Liu, C.-K. [Department of Chemical Engineering, National Tsing Hua University, Hsinchu 300 (China); Huang, T.-J. [Department of Chemical Engineering, National Tsing Hua University, Hsinchu 300 (China)]. E-mail: tjhuang@che.nthu.edu.tw

    2007-01-15

    The surface of porous three-dimensional (3D) calcium polyphosphate (CPP) scaffold was modified by treatment of quenching-after-sintering in the fabrication process. Scanning electron microscopic examination and degradation tests confirmed a new type of surface modification. A rotary-shaking culture was compared to that of a stationary culture and the results showed that rotary shaking led to enhanced extracellular matrices (ECM) secretion of both proteoglycans and collagen. Rotary-shaking cultured results showed that the quenching-treated CPP scaffold produced a better cartilage tissue, with both proteoglycans and collagen secretions enhanced, than the air-cooled-after-sintering scaffolds. Moreover, {beta}-CPP scaffolds were better for the ECM secretion of both proteoglycans and collagen than the {beta}-CPP + {gamma}-CPP multiphase scaffold. However, the multiphase scaffold led to higher growth rate than that of {beta}-CPP scaffold; the quenching-after-sintering treatment reversed this. In addition, the ECM secretions of both proteoglycans and collagen in the quenching-treated {beta}-CPP scaffold were higher than those in the air-cooled one. Thus, the novel treatment of quenching-after-sintering has shown merits to the porous 3D CPP scaffolds for articular cartilage tissue engineering.

  13. Inoculation of soil native cyanobacteria to restore arid degraded soils

    Science.gov (United States)

    Raúl Román Fernández, José; Roncero Ramos, Beatriz; Chamizo de la Piedra, Sonia; Rodríguez Caballero, Emilio; Ángeles Muñoz Martín, M.; Mateo, Pilar; Cantón Castilla, Yolanda

    2017-04-01

    Restoration projects in semiarid lands often yield poor results. Water scarcity, low soil fertility, and poor soil structure strongly limit the survival and growth of planted seedlings in these areas. Under these conditions, a previous stage that improves edaphic conditions would turn out to a successful plant restoration. By successfully colonizing arid soils, cyanobacteria naturally provide suitable edaphic conditions, enhancing water availability, soil fertility and soil stability. Furthermore, cyanobacteria can be easily isolated and cultured ex-situ to produce high quantities of biomass, representing a potential tool to restore large areas efficiently. The objective of this study was to test the effect of inoculated cyanobacteria on degraded soils at three different semiarid areas from southeast Spain: Tabernas badlands, a limestone quarry located in Gádor, and grazed grassland in Las Amoladeras (Cabo de Gata). Soil native cyanobacteria belonging to three representative N-fixing genera (Nostoc, Scytonema and Tolypothrix) were isolated from such soils and cultured in BG110 medium. Each strain was inoculated (6 g m-2), separately and mixed (all in the same proportion), on Petri dishes with 80 g of each soil. Biocrust development was monitored during 3 months in these soils under laboratory conditions, at a constant temperature of 25oC. During the experiment, two irrigation treatments were applied simulating a dry (180 mm) and a wet (360 mm) rainfall year (average recorded in the study sites). After 3 months, net CO2 flux, spectral response and soil surface microtopography (1 mm spatial resolution) of inoculated and control soils was measured under wet conditions, all of them as a surrogate of biocrust development. Samples of the surface crust were collected in order to determine total soil organic carbon (SOC) content. The inoculated soils showed positive values of net CO2 flux, thus indicating a net CO2 uptake, whereas control soils showed CO2 fluxes closed to

  14. Surface biofunctionalization of β-TCP blocks using aptamer 74 for bone tissue engineering

    Energy Technology Data Exchange (ETDEWEB)

    Ardjomandi, N.; Huth, J. [Department of Oral and Maxillofacial Surgery, University Hospital Tübingen (Germany); Stamov, D.R. [JPK Instruments AG, Berlin (Germany); Henrich, A. [Department of Oral and Maxillofacial Surgery, University Hospital Tübingen (Germany); Klein, C. [Dental Practice Zahngesundheit Waiblingen, Waiblingen (Germany); Wendel, H.-P. [Department of Thoracic, Cardiac and Vascular Surgery, University Hospital, Tübingen (Germany); Reinert, S. [Department of Oral and Maxillofacial Surgery, University Hospital Tübingen (Germany); Alexander, D., E-mail: dorothea.alexander@med.uni-tuebingen.de [Department of Oral and Maxillofacial Surgery, University Hospital Tübingen (Germany)

    2016-10-01

    Successful bone regeneration following oral and maxillofacial surgeries depends on efficient functionalization strategies that allow the recruitment of osteogenic progenitor cells at the tissue/implant interface. We have previously identified aptamer 74, which exhibited a binding affinity for osteogenically induced jaw periosteal cells (JPCs). In the present study, this aptamer was used for the surface biofunctionalization of β-tricalcium phosphate (β-TCP) blocks. Atomic force microscopy (AFM) measurements showed increased binding activity of aptamer 74 towards osteogenically induced JPCs compared to untreated controls. The immobilization efficiency of aptamer 74 was analyzed using the QuantiFluor ssDNA assay for 2D surfaces and by amino acid analysis for 3D β-TCP constructs. Following the successful immobilization of aptamer 74 in 2D culture wells and on 3D constructs, in vitro assays showed no significant differences in cell proliferation compared to unmodified surfaces. Interestingly, JPC mineralization was significantly higher on the 2D surfaces and higher cell adhesion was detected on the 3D constructs with immobilized aptamer. Herein, we report an established, biocompatible β-TCP matrix with surface immobilization of aptamer 74, which enhances properties such as cell adhesion on 3D constructs and mineralization on 2D surfaces. Further studies need to be performed to improve the immobilization efficiency and to develop a suitable approach for JPC mineralization growing within 3D β-TCP constructs. - Highlights: • Covalent binding of aptamer 74 on PLGA-coated β-tricalcium phosphate constructs. • AFM analysis of rupture forces between aptamer 74 and jaw periosteal cells. • Analysis of jaw periosteal cell functions on aptamer coated β-TCP constructs.

  15. A Comparison of the Reproduction of Surface Detail of Three Tissues Conditioning Materials over Time

    Directory of Open Access Journals (Sweden)

    Nili M.

    2012-09-01

    Full Text Available Statement of Problem: Although the basic use of tissue conditioners is to treat inflamed mucosa, they are also employed as functional impression materials. No information was obtained on the reproduction of surface detail of these materials over time.Purpose: The purpose of this study was to evaluate changes in the surface quality of three tissue conditioners after being immersed in water for a period of time.Materials and Method: Detail reproduction was determined by using from a ruled test block the same it was specified in ISO Specification 4823. Three tissue conditioners (GC, Acrosoft, and Visco-gel were evaluated. Samples were made by pouring freshly mixed materials into a ruled test block. The samples were then stored in distilled water for either of the followings periods of time: 0 hour, 24 hours, 3 days, 7 days, 14 days. Subsequently, the dental stone was mixed with distilled water and poured on each sample and allowed to remain for 60 minutes. Then, 25 specimens were prepared. The detail reproduction was determined based on what is specified in ISO Specification 4823. The samples were examined under a stereo-microscope with low-angle illumination. The data was analyzed through running Kruskal-Wallis Test and Mann-Whitney Test.Results: The three materials had minimum standard of detail reproduction. The detail reproduction were more significantly influenced by the time period of immersion in water ( p <0.0005 than the type of the material. The best detail reproduction was pertained to Visco-gel not immersed in water. Acrosoft was less influenced by the time period of immersion in water than the two other types.Conclusion: The detail reproduction may be attributed to chemical composition. The type of material and immersion time had a significant effect, while the effect of the type was less significant. The best time for making functional impressions was ranges from 0 to 24 hours.

  16. An Assessment of Cell Culture Plate Surface Chemistry for in Vitro Studies of Tissue Engineering Scaffolds

    Directory of Open Access Journals (Sweden)

    Alexander Röder

    2015-11-01

    Full Text Available The use of biopolymers as a three dimensional (3D support structure for cell growth is a leading tissue engineering approach in regenerative medicine. Achieving consistent cell seeding and uniform cell distribution throughout 3D scaffold culture in vitro is an ongoing challenge. Traditionally, 3D scaffolds are cultured within tissue culture plates to enable reproducible cell seeding and ease of culture media change. In this study, we compared two different well-plates with different surface properties to assess whether seeding efficiencies and cell growth on 3D scaffolds were affected. Cell attachment and growth of murine calvarial osteoblast (MC3T3-E1 cells within a melt-electrospun poly-ε-caprolactone scaffold were assessed when cultured in either “low-adhesive” non-treated or corona discharged-treated well-plates. Increased cell adhesion was observed on the scaffold placed in the surface treated culture plates compared to the scaffold in the non-treated plates 24 h after seeding, although it was not significant. However, higher cell metabolic activity was observed on the bases of all well-plates than on the scaffold, except for day 21, well metabolic activity was higher in the scaffold contained in non-treated plate than the base. These results indicate that there is no advantage in using non-treated plates to improve initial cell seeding in 3D polymeric tissue engineering scaffolds, however non-treated plates may provide an improved metabolic environment for long-term studies.

  17. Ocular surface reconstruction with a tissue-engineered nasal mucosal epithelial cell sheet for the treatment of severe ocular surface diseases.

    Science.gov (United States)

    Kobayashi, Masakazu; Nakamura, Takahiro; Yasuda, Makoto; Hata, Yuiko; Okura, Shoki; Iwamoto, Miyu; Nagata, Maho; Fullwood, Nigel J; Koizumi, Noriko; Hisa, Yasuo; Kinoshita, Shigeru

    2015-01-01

    Severe ocular surface diseases (OSDs) with severe dry eye can be devastating and are currently some of the most challenging eye disorders to treat. To investigate the feasibility of using an autologous tissue-engineered cultivated nasal mucosal epithelial cell sheet (CNMES) for ocular surface reconstruction, we developed a novel technique for the culture of nasal mucosal epithelial cells expanded ex vivo from biopsy-derived human nasal mucosal tissues. After the protocol, the CNMESs had 4-5 layers of stratified, well-differentiated cells, and we successfully generated cultured epithelial sheets, including numerous goblet cells. Immunohistochemistry confirmed the presence of keratins 3, 4, and 13; mucins 1, 16, and 5AC; cell junction and basement membrane assembly proteins; and stem/progenitor cell marker p75 in the CNMESs. We then transplanted the CNMESs onto the ocular surfaces of rabbits and confirmed the survival of this tissue, including the goblet cells, up to 2 weeks. The present report describes an attempt to overcome the problems of treating severe OSDs with the most severe dry eye by treating them using tissue-engineered CNMESs to supply functional goblet cells and to stabilize and reconstruct the ocular surface. The present study is a first step toward assessing the use of tissue-engineered goblet-cell transplantation of nonocular surface origin for ocular surface reconstruction.

  18. Anti-adhesion effects of liquid-infused textured surfaces on high-temperature stainless steel for soft tissue

    Science.gov (United States)

    Zhang, Pengfei; Chen, Huawei; Zhang, Liwen; Zhang, Deyuan

    2016-11-01

    Soft tissue adhesion on the electrosurgical instruments can induce many serious complications, such as failure of hemostasis and damage to the surrounding soft tissue. The soft tissue adhesion is mainly caused by the high temperature on the instrument surface generally made of stainless steel. Nepenthes inspired liquid-infused surfaces (LIS), highly promising for anti-adhesion, have attracted considerable interests. In this paper, we investigated the anti-adhesion effects of LIS on high-temperature stainless steel for soft tissue for the first time, aiming to develop a new approach to solve the soft tissue adhesion problem. The textured surface, acting as the holding structures, was fabricated by photolithography-assisted chemical etching. Silicone oil, with good biocompatibility and high-temperature resistance, was chosen as the infused liquid. The adhesion force measurements for soft tissue on the LIS at high temperatures indicated that the soft tissue adhesion force was decreased by approximately 80% at 250 °C. Besides, the cycle tests of soft tissue adhesion force demonstrated the excellent stability of prepared LIS. We anticipate that LIS will be of great promise for practical applications on the electrosurgical instruments.

  19. Experimental inoculation of domestic cats (Felis domesticus) with Sarcocystis neurona or S. neurona-like merozoites.

    Science.gov (United States)

    Butcher, M; Lakritz, J; Halaney, A; Branson, K; Gupta, G D; Kreeger, J; Marsh, A E

    2002-07-29

    Sarcocystis neurona is the parasite most commonly associated with equine protozoal myeloencephalitis (EPM). Recently, cats (Felis domesticus) have been demonstrated to be an experimental intermediate host in the life cycle of S. neurona. This study was performed to determine if cats experimentally inoculated with culture-derived S. neurona merozoites develop tissue sarcocysts infectious to opossums (Didelphis virginiana), the definitive host of S. neurona. Four cats were inoculated with S. neurona or S. neurona-like merozoites and all developed antibodies reacting to S. neurona merozoite antigens, but tissue sarcocysts were detected in only two cats. Muscle tissues from the experimentally inoculated cats with and without detectable sarcocysts were fed to laboratory-reared opossums. Sporocysts were detected in gastrointestinal (GI) scrapings of one opossum fed experimentally infected feline tissues. The study results suggest that cats can develop tissue cysts following inoculation with culture-derived Sarcocystis sp. merozoites in which the particular isolate was originally derived from a naturally infected cat with tissue sarcocysts. This is in contrast to cats which did not develop tissue cysts when inoculated with S. neurona merozoites originally derived from a horse with EPM. These results indicate present biological differences between the culture-derived merozoites of two Sarcocystis isolates, Sn-UCD 1 and Sn-Mucat 2.

  20. Tailoring surface topographies of polymers by using ion beam: Recent advances and the potential applications in biomedical and tissue engineering

    Energy Technology Data Exchange (ETDEWEB)

    Hasebe, Terumitsu, E-mail: teru_hasebe@sakura.med.toho-u.ac.jp [Department of Radiology, Toho University, Sakura Medical Center, 564-1 Shimoshizu, Sakura, Chiba 285-8741 (Japan); Center for Science of Environment, Resources and Energy, Graduate School of Science and Technology, Keio University, 3-14-1 Hiyoshi, Kohoku-ku, Yokohama, Kanagawa 223-8522 (Japan); Nagashima, So; Yoshimoto, Yukihiro; Hotta, Atsushi; Suzuki, Tetsuya [Center for Science of Environment, Resources and Energy, Graduate School of Science and Technology, Keio University, 3-14-1 Hiyoshi, Kohoku-ku, Yokohama, Kanagawa 223-8522 (Japan)

    2012-07-01

    Ion beam technique has recently been actively employed to create various patterns on the surface of polymers. In this paper, we highlight some of the recent advances in tailoring surface topographies of polymers by using ion beam and present a brief discussion on the potential applications in biomedical and tissue engineering.

  1. Changes in rumen bacterial community composition following feeding of silage inoculated with a commercial silage inoculant

    Science.gov (United States)

    Some silage inoculants yield an increase in milk production without increasing fiber digestibility, possibly through altering the rumen microflora. We hypothesized that silage treated with a commercial inoculant (Lactobacillus plantarum, LP) would improve milk production and would alter rumen bacter...

  2. Study on nasopharyngeal cancer tissue using surface-enhanced Raman spectroscopy

    Science.gov (United States)

    Ge, Xiaosong; Lin, Xueliang; Xu, Zhihong; Wei, Guoqiang; Huang, Wei; Lin, Duo

    2016-10-01

    Surface-enhanced Raman spectroscopy (SERS) can provide detailed molecular structure and composition information, and has demonstrated great potential in biomedical filed. This spectroscopy technology has become one of the most important optical techniques in the early diagnosis of cancer. Nasopharyngeal cancer (NPC) is a malignant neoplasm arising in the nasopharyngeal epithelial lining, which has relatively high incidence and death rate in Southeast Asia and southern China. This paper reviews the current progress of SERS in the field of cancer diagnostics, including gastric cancer, colorectal cancer, cervical cancer and nasopharyngeal cancer. In addition to above researches, we recently develop a novel NPC detection method based on tissue section using SERS, and obtain primary results. The proposed method has promising potential for the detection of nasopharyngeal carcinoma.

  3. Surface fluids effects on the bladder tissue characterisation using electrical impedance spectroscopy.

    Science.gov (United States)

    Keshtkar, Ahmad; Mesbahi, Asghar; Mehnati, Parinaz; Keshtkar, Asghar

    2008-07-01

    The electrical impedance of the human urinary bladder in both benign and malignant areas can be measured using an electrical impedance spectroscopy system (EIS). Glycine is usually used in the bladder surgery in the theatre to make an insulation medium for electro-surgery and the extension of the mucosa. In addition, a saline solution is usually used to wash the inside of the bladder after bladder surgery and it is used to extend the bladder tissue mucosa. Therefore, the effect of glycine and the saline solution that fills the bladder is important, because it was expected that the application of common surface fluids (air, saline solution and glycine solution) in the bladder epithelium would affect the measured electrical impedance of the urothelium, to differentiate the malignant area from the normal bladder tissue. In this study, bladders were removed from the patients' bodies and then were moved from theatre to the histopathology department immediately after excision. These bladder samples were then opened and pinned to a corkboard to take the impedance readings, using the impedance spectroscopy system. Following this, the bladder and corkboard were completely submerged in a saline solution and readings were taken at about 1cm from the sutures. Subsequently, this procedure was repeated with the bladder submerged in glycine and then air, respectively. According to the statistical work, these fluids were found to have a significant effect on the measured impedance of the bladder tissue in benign and malignant areas. Furthermore, the best fluid between air, glycine and saline, to measure the impedance of the urinary bladder, is air (P<0.0001).

  4. Fibroblast/fibrocyte: surface interaction dictates tissue reactions to micropillar implants.

    Science.gov (United States)

    Baker, David W; Liu, Xinchuan; Weng, Hong; Luo, Cheng; Tang, Liping

    2011-04-11

    Micropillar technology has shown great promise for medical implants or sensors in recent years. To study the influence of surface topography on cellular responses, polydimethylsiloxane (PDMS) micropillar arrays with pillar spacing (20-70 μm) and height (14-25 μm) have been fabricated. The influence of micropillar arrays on cellular behavior was tested both in vitro and in vivo. Interestingly, in vitro, we observe a distinct response for 3T3 fibroblasts and RAW 264.7 macrophages to the topographical cues tested. Attachment and proliferation of fibroblasts was substantially enhanced by increasing pillar height, whereas macrophage adherence is significantly diminished by reduced pillar spacing. When implanted in the subcutaneous cavity of BALB/c mice for 14 days, we find a prevailing trend with capsule cell density and capsule thickness increasing, as both pillar height and spacing rise. Collagen deposition and neoangiogenesis, two pivotal factors in granulation tissue maturation, are also observed to have a stronger response to the increase in both pillar height and spacing. In contradiction to our original hypothesis, we observed that fibroblasts rather than macrophages are a key contributor to the in vivo outcome of micropillar arrays. Investigation into fibroblast activation, however, revealed that recruited fibrocytes, rather than resident fibroblasts, correspond to the in vivo outcome. The results from this work support the critical and often overlooked role of fibrocytes in tissue response to biomaterial implants with varying topography.

  5. Surface fluorescence studies of tissue mitochondrial redox state in isolated perfused rat lungs.

    Science.gov (United States)

    Staniszewski, Kevin; Audi, Said H; Sepehr, Reyhaneh; Jacobs, Elizabeth R; Ranji, Mahsa

    2013-04-01

    We designed a fiber-optic-based optoelectronic fluorometer to measure emitted fluorescence from the auto-fluorescent electron carriers NADH and FAD of the mitochondrial electron transport chain (ETC). The ratio of NADH to FAD is called the redox ratio (RR = NADH/FAD) and is an indicator of the oxidoreductive state of tissue. We evaluated the fluorometer by measuring the fluorescence intensities of NADH and FAD at the surface of isolated, perfused rat lungs. Alterations of lung mitochondrial metabolic state were achieved by the addition of rotenone (complex I inhibitor), potassium cyanide (KCN, complex IV inhibitor) and/or pentachlorophenol (PCP, uncoupler) into the perfusate recirculating through the lung. Rotenone- or KCN-containing perfusate increased RR by 21 and 30%, respectively. In contrast, PCP-containing perfusate decreased RR by 27%. These changes are consistent with the established effects of rotenone, KCN, and PCP on the redox status of the ETC. Addition of blood to perfusate quenched NADH and FAD signal, but had no effect on RR. This study demonstrates the capacity of fluorometry to detect a change in mitochondrial redox state in isolated perfused lungs, and suggests the potential of fluorometry for use in in vivo experiments to extract a sensitive measure of lung tissue health in real-time.

  6. Inoculation of sugarcane with diazotrophic bacteria

    Directory of Open Access Journals (Sweden)

    Nivaldo Schultz

    2014-04-01

    Full Text Available The sugarcane industry, a strategic crop in Brazil, requires technological improvements in production efficiency to increase the crop energy balance. Among the various currently studied alternatives, inoculation with diazotrophic bacteria proved to be a technology with great potential. In this context, the efficiency of a mixture of bacterial inoculant was evaluated with regard to the agronomic performance and N nutrition of sugarcane. The experiment was carried out on an experimental field of Embrapa Agrobiologia, in Seropédica, Rio de Janeiro, using a randomized block, 2 × 3 factorial design (two varieties and three treatments with four replications, totaling 24 plots. The varieties RB867515 and RB72454 were tested in treatments consisting of: inoculation with diazotrophic bacteria, N-fertilized control with 120 kg ha-1 N and absolute control (no inoculation and no N fertilizer. The inoculum was composed of five strains of five diazotrophic species. The yield, dry matter accumulation, total N in the shoot dry matter and the contribution of N by biological fixation were evaluated, using the natural 15N abundance in non-inoculated sugarcane as reference. The bacterial inoculant increased the stalk yield of variety RB72454 similarly to fertilization with 120 kg ha-1 N in the harvests of plant-cane and first ratoon crops, however the contribution of biological N fixation was unchanged by inoculation, indicating that the benefits of the inoculant in sugarcane may have resulted from plant growth promotion.

  7. The influence of surface topography of a porous perfluoropolyether polymer on corneal epithelial tissue growth and adhesion.

    Science.gov (United States)

    Evans, Margaret D M; Chaouk, Hassan; Wilkie, John S; Dalton, Beatrice A; Taylor, Sarah; Xie, Ruo Zhong; Hughes, Timothy C; Johnson, Graham; McFarland, Gail A; Griesser, Hans H; Steele, John G; Meijs, Gordon F; Sweeney, Deborah F; McLean, Keith M

    2011-12-01

    Design principles for corneal implants are challenging and include permeability which inherently involves pore openings on the polymer surface. These topographical cues can be significant to a successful clinical outcome where a stratified epithelium is needed over the device surface, such as with a corneal onlay or corneal repair material. The impact of polymer surface topography on the growth and adhesion of corneal epithelial tissue was assessed using porous perfluoropolyether membranes with a range of surface topography. Surfaces were characterised by AFM and XPS, and the permeability and water content of membranes was measured. Biological testing of membranes involved a 21-day in vitro tissue assay to evaluate migration, stratification and adhesion of corneal epithelium. Similar parameters were monitored in vivo by surgically implanting membranes into feline corneas for up to 5 months. Data showed optimal growth and adhesion of epithelial tissue in vitro when polymer surface features were below a 150 nm RMS value. Normal processes of tissue growth and adhesion were disrupted when RMS values approached 300 nm. Data from the in vivo study confirmed these findings. Together, outcomes demonstrated the importance of surface topography in the design of implantable devices that depend on functional epithelial cover.

  8. Immunohistochemical detection of Clostridia species in paraffin-embedded tissues of experimentally inoculated guinea pigs Detecção imunohistoquímica de várias espécies de Clostridia em tecidos parafinizados de cobaias inoculadas experimentalmente

    Directory of Open Access Journals (Sweden)

    Ronnie A. Assis

    2005-03-01

    Full Text Available Blackleg is caused by Clostridium chauvoei, whereas malignant oedema is caused by C. chauvoei, C. septicum, C. sordellii, C. perfringens type A, and/or C. novyi type A. Anti-C. chauvoei, anti-C. septicum, anti-C. sordellii and anti-C. novyi type A polyclonal antibodies were produced in rabbits and purified in a column of DEAE-cellulose. Aliquots of the antisera were conjugated with fluorescein isothiocyanate and the remaining was used for the streptavidin biotin peroxidase technique (SBPT. SBPT was standardized to detect C. chauvoei, C. septicum, C. sordellii and C. novyi type A in formalin-fixed, paraffin-embedded tissues of guinea pigs. SBPT was compared to a fluorescent antibody technique (FAT. Sections and smears of muscle from inoculation area (MIA, heart, liver, spleen and kidney, were obtained for both SBPT and FAT. Cross-reactions between the different Clostridial species were not observed. C. chauvoei and C. septicum were detected in all specimens from the animals inoculated with these microorganisms, while only sections of muscle obtained from all the animals inoculated with C. sordellii and C. novyi type A were positive. The same results observed by the SBPT, were obtained on tissue smears of these microorganisms stained by the FAT. The results indicate that SBPT is suitable for detection of C. chauvoei, C. septicum, C. sordellii and C. novyi type A in formalin-fixed, paraffin-embedded tissues of guinea pigs.O carbúnculo sintomático é causado pelo Clostridium chauvoei, enquanto o edema maligno é causado pelo C. chauvoei, C. septicum, C. sordellii, C. perfringens tipo A, e/ou C. novyi tipo A. Corpos policlonais anti-C. chauvoei, anti-C. septicum, anti-C. sordellii and anti-C.novyi tipo A foram produzidos em coelhos e purificados em uma coluna de DEAE-celulose. Alíquotas das imunoglobulinas foram conjugadas com isotiocianato de fluoresceína e o restante foi usado na técnica de streptavidina biotina peroxidase (SBP. SBP foi

  9. Liquid Microjunction Surface Sampling Probe Electrospray Mass Spectrometry for Detection of Drugs and Metabolites in Thin Tissue Sections

    Energy Technology Data Exchange (ETDEWEB)

    Van Berkel, Gary J [ORNL; Kertesz, Vilmos [ORNL; Koeplinger, Kenneth A. [Merck Research Laboratories; Vavek, Marissa [Merck Research Laboratories; Kong, Ah-Ng Tony [Rutgers University

    2008-01-01

    A self-aspirating, liquid micro-junction surface sampling probe/electrospray emitter mass spectrometry system was demonstrated for use in the direct analysis of spotted and dosed drugs and their metabolites in thin tissue sections. Proof-of-principle sampling and analysis directly from tissue without the need for sample preparation was demonstrated first by raster scanning a region on a section of rat liver onto which reserpine was spotted. The mass spectral signal from selected reaction monitoring was used to develop a chemical image of the spotted drug on the tissue. The probe was also used to selectively spot sample areas of sagittal whole mouse body tissue sections that had been dosed orally (90 mg/kg) with R,S-sulforaphane 3 hrs prior to sacrifice. Sulforaphane and its glutathione and N-acetyl cysteine conjugates were monitored with selected reaction monitoring and detected in the stomach and various other tissues from the dosed mouse. No signal for these species was observed in the tissue from a control mouse. The same dosed tissue section was used to illustrate the possibility of obtaining a line scan across the whole body section. In total these results illustrate the potential for rapid screening of the distribution of drugs and metabolites in tissue sections with the micro-liquid junction surface sampling probe/electrospray mass spectrometry approach.

  10. Solvent Separating Secondary Metabolites Directly from Biosynthetic Tissue for Surface-Assisted Laser Desorption Ionisation Mass Spectrometry

    Directory of Open Access Journals (Sweden)

    David Rudd

    2015-03-01

    Full Text Available Marine bioactive metabolites are often heterogeneously expressed in tissues both spatially and over time. Therefore, traditional solvent extraction methods benefit from an understanding of the in situ sites of biosynthesis and storage to deal with heterogeneity and maximize yield. Recently, surface-assisted mass spectrometry (MS methods namely nanostructure-assisted laser desorption ionisation (NALDI and desorption ionisation on porous silicon (DIOS surfaces have been developed to enable the direct detection of low molecular weight metabolites. Since direct tissue NALDI-MS or DIOS-MS produce complex spectra due to the wide variety of other metabolites and fragments present in the low mass range, we report here the use of “on surface” solvent separation directly from mollusc tissue onto nanostructured surfaces for MS analysis, as a mechanism for simplifying data annotation and detecting possible artefacts from compound delocalization during the preparative steps. Water, ethanol, chloroform and hexane selectively extracted a range of choline esters, brominated indoles and lipids from Dicathais orbita hypobranchial tissue imprints. These compounds could be quantified on the nanostructured surfaces by comparison to standard curves generated from the pure compounds. Surface-assisted MS could have broad utility for detecting a broad range of secondary metabolites in complex marine tissue samples.

  11. Surface chemical immobilization of bioactive peptides on synthetic polymers for cardiac tissue engineering.

    Science.gov (United States)

    Rosellini, Elisabetta; Cristallini, Caterina; Guerra, Giulio D; Barbani, Niccoletta

    2015-01-01

    The aim of this work was the development of new synthetic polymeric systems, functionalized by surface chemical modification with bioactive peptides, for myocardial tissue engineering. Polycaprolactone and a poly(ester-ether-ester) block copolymer synthesized in our lab, polycaprolactone-poly(ethylene oxide)-polycaprolactone (PCL-PEO-PCL), were used as the substrates to be modified. Two pentapeptides, H-Gly-Arg-Gly-Asp-Ser-OH (GRGDS) from fibronectin and H-Tyr-Ile-Gly-Ser-Arg-OH (YIGSR) from laminin, were used for the functionalization. Polymeric membranes were obtained by casting from solutions and then functionalized by means of alkaline hydrolysis and subsequent coupling of the bioactive molecules through 1-(3-dimethylaminopropyl)-3-ethylcarbodimide hydrochloride/N-hydroxysuccinimide chemistry. The hydrolysis conditions, in terms of hydrolysis time, temperature, and sodium hydroxide concentration, were optimized for the two materials. The occurrence of the coupling reaction was demonstrated by infrared spectroscopy, as the presence on the functionalized materials of the absorption peaks typical of the two peptides. The peptide surface density was determined by chromatographic analysis and the distribution was studied by infrared chemical imaging. The results showed a nearly homogeneous peptide distribution, with a density above the minimum value necessary to promote cell adhesion. Preliminary in vitro cell culture studies demonstrated that the introduction of the bioactive molecules had a positive effect on improving C2C12 myoblasts growth on the synthetic materials.

  12. Developmentally regulated epitopes of cell surface arabinogalactan proteins and their relation to root tissue pattern formation.

    Science.gov (United States)

    Knox, J P; Linstead, P J; Peart, J; Cooper, C; Roberts, K

    1991-11-01

    Two polymorphic forms of an extracellular arabinogalactan protein (AGP1 and AGP2), obtained from the conditioned media of two carrot suspension-cultured cell lines, have been identified in terms of binding of the anti-plasma membrane antibodies JIM4 and MAC207. AGP1 and AGP2 have been used as immunogens to generate further anti-AGP monoclonal antibodies. JIM14 identified an epitope carried by AGP2 and also by glycoproteins of low molecular weight localized to the plant cell wall. In addition, further antibodies (JIM13 and JIM15) identified carbohydrate epitopes of the AGPs that also occur on plasma membrane glycoproteins and are expressed by patterns of cells that reflect cell position at the carrot root apex. Indirect immunofluorescence microscopy indicated that JIM13 recognized the surface of cells forming the epidermis and cells marking the region and axis of the future xylem. JIM15 recognized a pattern of cells directly complementary to the JIM13 pattern. The panel of anti-AGP monoclonal antibodies now available indicates groups of cells within the root meristem that may reflect an early pre-pattern of the tissues of the mature root structure and suggests extensive modulation of cell surface AGPs during cell development and the positioning of cells within the apex.

  13. Pneumonia in mice inoculated experimentally with Acanthamoeba polyphaga mimivirus.

    Science.gov (United States)

    Khan, Mohammad; La Scola, Bernard; Lepidi, Hubert; Raoult, Didier

    2007-01-01

    In an attempt to elucidate whether Acanthamoeba polyphaga mimivirus (APM) was pathogenic (i.e., induces histological evidence of inflammation or tissue destruction or both) in mammalian host, we inoculated adult BALB/c (n=6) and C57BL/6 (n=6) mice with APM via intracardiac route. C57BL/6 mice developed histopathological features of pneumonia by post-inoculation day (PID) 3 and BALB/c mice, by PID7. The histopathological features of pneumonia, characterized by the presence of thickened alveolar walls with cellular infiltrates comprising mononuclear leukocytes, macrophages and lymphocytes, diffuse alveolar damages with the formation of hyaline membrane and erythrocytes in the alveolar lumen, were most marked by PID 7. We conclude that APM is pathogenic in mice in experimental conditions.

  14. Experimental Inoculation of Egyptian Fruit Bats (Rousettus aegyptiacus) with Ebola Virus.

    Science.gov (United States)

    Paweska, Janusz T; Storm, Nadia; Grobbelaar, Antoinette A; Markotter, Wanda; Kemp, Alan; Jansen van Vuren, Petrus

    2016-01-22

    Colonized Egyptian fruit bats (Rousettus aegyptiacus), originating in South Africa, were inoculated subcutaneously with Ebola virus (EBOV). No overt signs of morbidity, mortality, or gross lesions were noted. Bats seroconverted by Day 10-16 post inoculation (p.i.), with the highest mean anti-EBOV IgG level on Day 28 p.i. EBOV RNA was detected in blood from one bat. In 16 other tissues tested, viral RNA distribution was limited and at very low levels. No seroconversion could be demonstrated in any of the control bats up to 28 days after in-contact exposure to subcutaneously-inoculated bats. The control bats were subsequently inoculated intraperitoneally, and intramuscularly with the same dose of EBOV. No mortality, morbidity or gross pathology was observed in these bats. Kinetics of immune response was similar to that in subcutaneously-inoculated bats. Viral RNA was more widely disseminated to multiple tissues and detectable in a higher proportion of individuals, but consistently at very low levels. Irrespective of the route of inoculation, no virus was isolated from tissues which tested positive for EBOV RNA. Viral RNA was not detected in oral, nasal, ocular, vaginal, penile and rectal swabs from any of the experimental groups.

  15. Klebsiella pneumoniae inoculants for enhancing plant growth

    Energy Technology Data Exchange (ETDEWEB)

    Triplett, Eric W. (Middleton, WI); Kaeppler, Shawn M. (Oregon, WI); Chelius, Marisa K. (Greeley, CO)

    2008-07-01

    A biological inoculant for enhancing the growth of plants is disclosed. The inoculant includes the bacterial strains Herbaspirillum seropedicae 2A, Pantoea agglomerans P101, Pantoea agglomerans P102, Klebsiella pneumoniae 342, Klebsiella pneumoniae zmvsy, Herbaspirillum seropedicae Z152, Gluconacetobacter diazotrophicus PA15, with or without a carrier. The inoculant also includes strains of the bacterium Pantoea agglomerans and K. pneumoniae which are able to enhance the growth of cereal grasses. Also disclosed are the novel bacterial strains Herbaspirillum seropedicae 2A, Pantoea agglomerans P101 and P102, and Klebsiella pneumoniae 342 and zmvsy.

  16. Klebsiella pneumoniae inoculants for enhancing plant growth

    Science.gov (United States)

    Triplett, Eric W [Middleton, WI; Kaeppler, Shawn M [Oregon, WI; Chelius, Marisa K [Greeley, CO

    2008-07-01

    A biological inoculant for enhancing the growth of plants is disclosed. The inoculant includes the bacterial strains Herbaspirillum seropedicae 2A, Pantoea agglomerans P101, Pantoea agglomerans P102, Klebsiella pneumoniae 342, Klebsiella pneumoniae zmvsy, Herbaspirillum seropedicae Z152, Gluconacetobacter diazotrophicus PA15, with or without a carrier. The inoculant also includes strains of the bacterium Pantoea agglomerans and K. pneumoniae which are able to enhance the growth of cereal grasses. Also disclosed are the novel bacterial strains Herbaspirillum seropedicae 2A, Pantoea agglomerans P101 and P102, and Klebsiella pneumoniae 342 and zmvsy.

  17. A thermal monitoring sheet with low influence from adjacent waterbolus for tissue surface thermometry during clinical hyperthermia.

    Science.gov (United States)

    Arunachalam, Kavitha; Maccarini, Paolo F; Stauffer, Paul R

    2008-10-01

    This paper presents a complete thermal analysis of a novel conformal surface thermometer design with directional sensitivity for real-time temperature monitoring during hyperthermia treatments of large superficial cancer. The thermal monitoring sheet (TMS) discussed in this paper consists of a 2-D array of fiberoptic sensors embedded between two layers of flexible, low-loss, and thermally conductive printed circuit board (PCB) film. Heat transfer across all interfaces from the tissue surface through multiple layers of insulating dielectrics surrounding the small buried temperature sensor and into an adjacent temperature-regulated water coupling bolus was studied using 3-D thermal simulation software. Theoretical analyses were carried out to identify the most effective differential TMS probe configuration possible with commercially available flexible PCB materials and to compare their thermal responses with omnidirectional probes commonly used in clinical hyperthermia. A TMS sensor design that employs 0.0508-mm Kapton MTB and 0.2032-mm Kapton HN flexible polyimide films is proposed for tissue surface thermometry with low influence from the adjacent waterbolus. Comparison of the thermal simulations with clinical probes indicates the new differential TMS probe design to outperform in terms of both transient response and steady-state accuracy in selectively reading the tissue surface temperature, while decreasing the overall thermal barrier of the probe between the coupling waterbolus and tissue surface.

  18. Highly bioactive polysiloxane modified bioactive glass-poly(ethylene glycol) hybrids monoliths with controlled surface structure for bone tissue regeneration

    Science.gov (United States)

    Chen, Jing; Que, Wenxiu; Xing, Yonglei; Lei, Bo

    2015-03-01

    Crack-free monoliths with controllable surface microstructure have high bioactivities and therefore potential applications in bone tissue regeneration. In this paper, crack-free polydimethylsiloxane-modified bioactive glass-poly (ethylene glycol) (PDMS-BG-PEG) hybrids monoliths were fabricated via using a modified sol-gel process. Results show that the addition of PEG plays an important part in the formation of crack-free and gelation of the monoliths, and surface microstructures of the as-prepared hybrid monoliths were significantly influenced by the concentration and molecular weight of PEG. The samples obtained from PEG 300 had porous surface result in higher bioactivity (apatite formation) in simulated body fluid (SBF), while the samples obtained from PEG 600 had the smooth surface and inhibited the formation of apatite layer in SBF. These as-prepared hybrid monoliths can be used as a good candidate of implant and scaffold for highly efficient bone tissue regeneration.

  19. Tissue factor expression on the surface of monocytes from a patient with hereditary angioedema.

    Science.gov (United States)

    Iwamoto, Kazumasa; Morioke, Satoshi; Yanase, Yuhki; Uchida, Kazue; Hide, Michihiro

    2014-10-01

    Hereditary angioedema (HAE) presents as severe angioedema, which is mostly due to the C1 inhibitor (C1-INH) gene mutations. Environmental factors, minor trauma and oral contraceptives have been reported to induce angioedema attack, but the trigger may often be uncertain. Activated factor XII controlled by C1-INH facilitates bradykinin generation and also regulates coagulation cascade, but the relationship between edema formation and coagulation is still unclear. We have described a 35-year-old female patient with HAE, presenting with frequent angioedema attacks in the absence of an apparent triggering factor. She showed higher levels of FDP and D-dimer during angioedema than those in remission. In addition, tissue factor (TF), an initiator of the extrinsic coagulation cascade, was expressed on the surface of monocytes. It was significantly higher than that of monocytes from healthy controls and tends to further increase during attacks. The expression of TF on monocytes may play a role in the induction of angioedema attacks in HAE by activating the coagulation pathway in association with reduced functions of C1-INH.

  20. Simulation study of melanoma detection in human skin tissues by laser-generated surface acoustic waves

    Science.gov (United States)

    Chen, Kun; Fu, Xing; Dorantes-Gonzalez, Dante J.; Lu, Zimo; Li, Tingting; Li, Yanning; Wu, Sen; Hu, Xiaotang

    2014-07-01

    Air pollution has been correlated to an increasing number of cases of human skin diseases in recent years. However, the investigation of human skin tissues has received only limited attention, to the point that there are not yet satisfactory modern detection technologies to accurately, noninvasively, and rapidly diagnose human skin at epidermis and dermis levels. In order to detect and analyze severe skin diseases such as melanoma, a finite element method (FEM) simulation study of the application of the laser-generated surface acoustic wave (LSAW) technique is developed. A three-layer human skin model is built, where LSAW's are generated and propagated, and their effects in the skin medium with melanoma are analyzed. Frequency domain analysis is used as a main tool to investigate such issues as minimum detectable size of melanoma, filtering spectra from noise and from computational irregularities, as well as on how the FEM model meshing size and computational capabilities influence the accuracy of the results. Based on the aforementioned aspects, the analysis of the signals under the scrutiny of the phase velocity dispersion curve is verified to be a reliable, a sensitive, and a promising approach for detecting and characterizing melanoma in human skin.

  1. The ratio of the spherical and flat Detectors at tissue surfaces during pleural photodynamic therapy.

    Science.gov (United States)

    Zhu, Timothy C; Friedberg, Joseph S; Dimofte, Andrea; Miles, Jeremy; Metz, James; Glatstein, Eli; Hahn, Stephen M

    2002-06-06

    An isotropic detector-based system was compared with a flat photodiode-based system in patients undergoing pleural photodynamic therapy. Isotropic and flat detectors were placed side by side in the chest cavity, for simultaneous in vivo dosimetry at surface locations for twelve patients. The treatment used 630nm laser to a total light irradiance of 30 J/cm(2) (measured with the flat photodiodes) with photofrin® IV as the photosensitizer. Since the flat detectors were calibrated at 532nm, wavelength correction factors (WCF) were used to convert the calibration to 630nm (WCF between 0.542 and 0.703). The mean ratio between isotropic and flat detectors for all sites was linear to the accumulated fluence and was 3.4±0.6 or 2.1±0.4, with or without the wavelength correction for the flat detectors, respectively. The μeff of the tissues was estimated to vary between 0.5 to 4.3 cm(-1) for four sites (Apex, Posterior Sulcus, Anterior Chest Wall, and Posterior Mediastinum) assuming μs' = 7 cm(-1). Insufficient information was available to estimate μeff directly for three other sites (Anterior Sulcus, Posterior Chest Wall, and Pericardium) primarily due to limited sample size, although one may assume the optical penetration in all sites to vary in the same range (0.5 to 4.3 cm(-1)).

  2. Experimental Transmission of the Chronic Wasting Disease Agent to Swine after Oral or Intracranial Inoculation.

    Science.gov (United States)

    Moore, S Jo; West Greenlee, M Heather; Kondru, Naveen; Manne, Sireesha; Smith, Jodi D; Kunkle, Robert A; Kanthasamy, Anumantha; Greenlee, Justin J

    2017-10-01

    Chronic wasting disease (CWD) is a naturally occurring, fatal neurodegenerative disease of cervids. The potential for swine to serve as hosts for the agent of CWD is unknown. The purpose of this study was to investigate the susceptibility of swine to the CWD agent following experimental oral or intracranial inoculation. Crossbred piglets were assigned to three groups, intracranially inoculated (n = 20), orally inoculated (n = 19), and noninoculated (n = 9). At approximately the age at which commercial pigs reach market weight, half of the pigs in each group were culled ("market weight" groups). The remaining pigs ("aged" groups) were allowed to incubate for up to 73 months postinoculation (mpi). Tissues collected at necropsy were examined for disease-associated prion protein (PrP(Sc)) by Western blotting (WB), antigen capture enzyme immunoassay (EIA), immunohistochemistry (IHC), and in vitro real-time quaking-induced conversion (RT-QuIC). Brain samples from selected pigs were also bioassayed in mice expressing porcine prion protein. Four intracranially inoculated aged pigs and one orally inoculated aged pig were positive by EIA, IHC, and/or WB. By RT-QuIC, PrP(Sc) was detected in lymphoid and/or brain tissue from one or more pigs in each inoculated group. The bioassay was positive in four out of five pigs assayed. This study demonstrates that pigs can support low-level amplification of CWD prions, although the species barrier to CWD infection is relatively high. However, detection of infectivity in orally inoculated pigs with a mouse bioassay raises the possibility that naturally exposed pigs could act as a reservoir of CWD infectivity.IMPORTANCE We challenged domestic swine with the chronic wasting disease agent by inoculation directly into the brain (intracranially) or by oral gavage (orally). Disease-associated prion protein (PrP(Sc)) was detected in brain and lymphoid tissues from intracranially and orally inoculated pigs as early as 8 months of age (6 months

  3. Inoculation of chromium white cast iron

    Directory of Open Access Journals (Sweden)

    D. Kopyciński

    2009-01-01

    Full Text Available It has been proved that an addition of boron carbide introduced as an inoculant to the chromium white cast iron changes the structureof castings. Castings after inoculation revealed a different structure with numerous grains. Primary precipitates of chromium carbide also appeared, reducing the mechanical properties of as-cast parts. Properly established heat treatment regime makes chromium iron castings regain their, originally high, mechanical properties.

  4. New inoculants on maize silage fermentation

    Directory of Open Access Journals (Sweden)

    Fábia Giovana do Val de Assis

    2014-08-01

    Full Text Available The objective of this study was to evaluate the effect of bacterial inoculants at two inoculation rates on chemical and biological characteristics of maize silage. The treatments consisted of two inoculating rates (5 and 6 log cfu g-1 of forage for each strain of lactic acid bacteria (LAB identified as Lactobacillus buchneri, L. hilgardii, or L. plantarum. The maize was ensiled in experimental PVC silos. Samples were taken for the determination of the contents of dry matter (DM, crude protein (CP, neutral detergent fiber (NDF, water-soluble carbohydrates (WSC, organic acids and alcohols, for the evaluation of the populations of lactic acid bacteria, yeasts, filamentous fungi, and for the determination of pH values during ensilage and after 30 or 90 days of fermentation. The doses of inoculants did not promote significant differences on the evaluated characteristics. There was effect of inoculants on acetic acid, 1.2-propanediol, LAB population, filamentous fungi, and pH value. No significant influence of the treatments with inoculants was observed in the variables DM, WSC, CP, lactic acid concentrations, or ethanol. The maximum temperature, i.e., the time to achieve the maximum temperature (TMT and aerobic stability (AS, was not influencied by treatments. However, a decrease in maximum temperature, an increase in TMT, and improvement in the AS were observed after 90 days of fermentation. These results proved the advantage of microbial inoculation. The treatments influenced LAB populations and filamentous fungi, but no effect was observed on the yeast population. The best inoculation dose is 6 cfu g-1 of forage because it provides higher reduction of filamentous fungi in maize silage, thereby decreasing the aerobic deterioration by these microorganisms.

  5. Surface modification by plasma etching impairs early vascularization and tissue incorporation of porous polyethylene (Medpor(®) ) implants.

    Science.gov (United States)

    Laschke, Matthias W; Augustin, Victor A; Sahin, Fadime; Anschütz, Dieter; Metzger, Wolfgang; Scheuer, Claudia; Bischoff, Markus; Aktas, Cenk; Menger, Michael D

    2016-11-01

    Porous polyethylene (Medpor®) is commonly used in craniofacial reconstructive surgery. Rapid vascularization and tissue incorporation are crucial for the prevention of migration, extrusion, and infection of the biomaterial. Therefore, we analyzed whether surface modification by plasma etching may improve the early tissue response to Medpor®. Medpor® samples were treated in a plasma chamber at low (20 W; LE-PE) and high energy levels (40 W; HE-PE). The samples and non-treated controls were implanted into mouse dorsal skinfold chambers to analyze angiogenesis, inflammation, and granulation tissue formation over 14 days using intravital fluorescence microscopy, histology, and immunohistochemistry. Scanning electron microscopy (SEM) analyses revealed that elevating energy levels of plasma etching progressively increase the oxygen surface content and surface roughness of Medpor®. This did not affect the leukocytic response to the implants. However, LE-PE and HE-PE samples exhibited an impaired vascularization. This was associated with a reduced formation of a collagen-rich granulation tissue at the implantation site. Additional in vitro experiments showed a reduced cell attachment on plasma-etched Medpor®. Thus, plasma etching may not be recommended to improve the clinical outcome of reconstructive interventions using Medpor®. However, it may be beneficial for temporarily implanted polyethylene-based biomedical devices for which tissue incorporation is undesirable. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 104B: 1738-1748, 2016. © 2015 Wiley Periodicals, Inc.

  6. A ‘tissue model’ to study the barrier effects of living tissues on the reactive species generated by surface air discharge

    Science.gov (United States)

    He, Tongtong; Liu, Dingxin; Xu, Han; liu, Zhichao; Xu, Dehui; Li, Dong; Li, Qiaosong; Rong, Mingzhe; Kong, Michael G.

    2016-05-01

    Gelatin gels are used as surrogates of human tissues to study their barrier effects on incoming reactive oxygen and nitrogen species (RONS) generated by surface air discharge. The penetration depth of nitrite into gelatin gel is measured in real time during plasma treatment, and the permeabilities of nitrite, nitrate, O3 and H2O2 through gelatin gel films are quantified by measuring their concentrations in the water underneath such films after plasma treatment. It is found that the penetration speed of nitrite increases linearly with the mass fraction of water in the gelatin gels, and the permeabilities of nitrite and O3 are comparably smaller than that for H2O2 and nitrate due to differences in their chemistry in gelatin gels. These results provide a quantitative basis to estimate the penetration processes of RONS in human tissues, and they also confirm that the composition of RONS is strongly dependent on the tissue depth and the plasma treatment time. A small electric field of up to 20 V cm-1 can greatly reduce the barrier effects of the tissue model regardless of their directions, for which the underlying mechanism is unclear. However, the electric field force on the objective RONS should not be the dominant mechanism.

  7. La inoculación de plantas con Pantoea sp., bacteria solubilizadora de fosfatos, incrementa la concentración de P en los tejidos foliares Plant inoculation with Pantoea sp., phosphate solubilising-bacteria increases P concentration in leaf tissues

    Directory of Open Access Journals (Sweden)

    Ortega-Rodés Patricia

    2008-07-01

    inoculants for crops is a biotechnological alternative to increasing its availability. Pantoea sp. (9C strain is an endophytic nitrogen-fixing bacterium isolated from the inside of sugarcane stems (Loiret et ál., 2004. The microorganism is able to produce 6 mm solubilisation halo in plate assays after 7 days at 30ºC, and also solubilise Ca3(PO42 in NBRI-P liquid medium, producing values of 1,128 μg P mL-1. It is also able to survive for at least 35 days in a substrate mixture prepared with Vermiculite and Red Ferralitic (Ferralic Cambisol, rhodic soil, producing populations of 3.2 x 1,015 cells g-1. Fast growth and high demanding P radish plants (Raphanus sativus, L. var. Scarlet Globe used as model plants cultivated in soil inoculated with the microorganism absorbed more P than the non-inoculated plants, reaching ≥ 3,500 ppm Pdry weight in foliar tissue. Key words: Endophytic bacteria; phosphate solubilization; radish (Raphanus sativus, L.; sugarcane (Saccharum hybrid.

  8. Research on the best measurement situation between optical probe and tissue surfaces in non-invasive detection

    Science.gov (United States)

    Yu, Xuyao; Liu, Rong; Yu, Hui; Wang, Jiao; Wang, Jun; Xu, Kexin

    2016-11-01

    Near-infrared spectroscopy is often used for the non-invasive detection of composition in the human body, such as that of blood glucose and haemoglobin, due to its high penetration depth into tissues. Although it is feasible to position the optical probe precisely, contact situation between probe and human tissues is a difficult problem to determine because of physiological tremor and mechanical performance of bio-soft tissue. Here, we proposed a novel estimation method for the situation between the optical probe and tissue surfaces based on the dynamic auto-correlation matrix of two-dimensional correlation spectroscopy (2DCOS) and radar chart. The diffuse reflectance spectra from the left palm of 4 healthy volunteers were collected while the optical probe gradually approached and pressed bio-tissues with a custom-design controlling device. 2DCOS in the wavelength with lower absorption (1000-1400 nm) was calculated under the perturbation of relative-distance and contact pressure between the optical probe and tissue surface. The synchronous 2DCOS showed that the surface reflection and diffuse reflectance were greatly affected by the contact conditions in 1100 nm, 1220 nm, and 1300 nm. Then the dynamic auto-correlation matrix of 2DCOS was established for the adjacent spectra, and the significant difference wavelengths were used to build radar charts to determine the critical contact situation visually. Results showed that the maximum variations of dynamic auto-correlation matrix appeared at near 1300 nm, and the relative distance between the probe and tissue corresponding to the critical contact state can be easily observed with radar charts with 0.25 mm uncertainty, which was consistent with the self-feeling of each volunteer. So this method can be applied to exactly determine the optimal measurement status for the non-invasive body composition detection in vivo. It is important for the design of human-machine interface and the accuracy improvement of body

  9. Comparison of the Surface Roughness of Gypsum (Dental Stone with three Types of Tissue Conditioner Impression Materials over Time

    Directory of Open Access Journals (Sweden)

    Nili M.

    2012-02-01

    Full Text Available Statement of Problems: Although the primary use of tissue conditioners is for healing the damaged mucosa but they can also be used as functional impression; it seems that its effeicasy depends on its viscoelastic features such as compatibility with gypsum and surface roughness.Purpose: The aim of this study is to compare the surface roughness of gypsum with several tissue conditioner impression materials avaliable in the market.Materials and Method: In this experimental study, three tissue conditioners (Acrosoft, viscogel & GC were used. Pars dental gypsum moldano Type III and a polyvinyl siloxane impression were used for the controls. The tissue conditioners powder liquid ratio was mixed according to the manufacturer’s recommendation and immediately poured in a mold with an internal diameter of 18 mm and depth of 2mm. The mold was completely filled. Then, a glass block with the mean roughness of 0.8 µm was placed on its surface for two hours. Then, the 5 samples were immediately placed in 37oC water for 0-24 hrs, 3, 7, and 14 days. After that, the specimens were beaded, boxed and poured with pars dental gypsum type III. The gypsum sample’s surface roughness was measured with profilometer with the length of 2.5 mm and cut-off of 0.8 mm. The results were analyzed using ANOVA and Tukey tests.Results: The results showed that surface roughness of Acrosoft in different storage times did not change significantly and there was no significant difference between Acrosoft and the control group. Viscogels surface roughness was significantly different with all other groups at zero time; with the increase of storage time the surface roughness decreases. The control group showed a significant difference with viscogel at zero time and with GC at 24 hrs and 3 days but it revealed no difference with the other groups. The least surface roughness belonged to GC at zero and 14 days and the highests surface roughness belonged to viscogel at zero time

  10. Research trends in biomimetic medical materials for tissue engineering: 3D bioprinting, surface modification, nano/micro-technology and clinical aspects in tissue engineering of cartilage and bone.

    Science.gov (United States)

    Chen, Cen; Bang, Sumi; Cho, Younghak; Lee, Sahnghoon; Lee, Inseop; Zhang, ShengMin; Noh, Insup

    2016-01-01

    This review discusses about biomimetic medical materials for tissue engineering of bone and cartilage, after previous scientific commentary of the invitation-based, Korea-China joint symposium on biomimetic medical materials, which was held in Seoul, Korea, from October 22 to 26, 2015. The contents of this review were evolved from the presentations of that symposium. Four topics of biomimetic medical materials were discussed from different research groups here: 1) 3D bioprinting medical materials, 2) nano/micro-technology, 3) surface modification of biomaterials for their interactions with cells and 4) clinical aspects of biomaterials for cartilage focusing on cells, scaffolds and cytokines.

  11. STUDY ON NUDE MICE INOCULATED WITH MYCOBACTERIUM LEPRAE BY MULTIPLE ROUTES

    Institute of Scientific and Technical Information of China (English)

    王荷英; 张伟云; 喻林冲; 施美琴; 刘季和

    1995-01-01

    Immune-defieient nude mice were inoculated with nude mouse derived Mycobacterium leprae by multiple routes (intravenously, subcutaneously at the foot pads and ears). The results showed that these inoculated animals were capable of producing a great number of Mycohecteritma leprae to a level 1011-12 per gram of tissue, and were detected histopathologically to have heavy lepromatous lesions. The dissemination of the infeetion was found particulerly in sites with lower body temperature. The ccgaulsms have a partiality to striated muscles and peripheral nerves. The authors suggest that experimental lepeosy in nude mice is a very useful tool for leprosy resarch, especially Jn cotmtries without armadillos. Compared with the single-route inoculation reported previously, multiple-route inoculation is more available.

  12. The effect of microstructured surfaces and laminin-derived peptide coatings on soft tissue interactions with titanium dental implants.

    Science.gov (United States)

    Werner, Sandra; Huck, Olivier; Frisch, Benoît; Vautier, Dominique; Elkaim, René; Voegel, Jean-Claude; Brunel, Gérard; Tenenbaum, Henri

    2009-04-01

    In the present study, we investigated the dental implant protection from peri-implant inflammation by improving the soft tissue adhesion on the titanium surface. Porous titanium was used to create, at the level of the transmucosal part of the implants (the "neck"), a microstructured 3-dimensional surface that would tightly seal the interface between the implant and soft tissue. Cell-specific adhesion properties were induced via an adhesion peptide derived from laminin-5 coupled to native or cross-linked PLL/PGA multilayered polyelectrolyte films (MPFs), which are used for biomedical device coatings. Porous titanium exhibited good cell-adhesion properties, but the colonisation of the material was further improved by a coating with laminin-5 functionalised MPFs and especially with (PLL/PGA)(6,5)-PGA-peptide film. Focal contact formation was observed on cross-linked architectures, reflecting cell anchorage on these surfaces. In contrast, when seeded on laminin-5-functionalised native films, epithelial cells formed only very diffuse focal contacts, but adhered via hemidesmosome formation. In vivo experiments confirmed that the porous titanium was colonised by cells of soft tissue. Altogether, the results indicate that the microstructure of the implant neck combined with a specific bioactive coating could constitute efficient routes to improve the integration of soft tissue on titanium dental implants, which could significantly protect implants from peri-implant inflammation and enhance long-term implant stabilisation.

  13. Invasive facial fungal infections: Orofacial soft-tissue infiltration in immunocompromised patients

    OpenAIRE

    Jun, Peter; Russell, Matthew; El-Sayed, Ivan; Dillon, William; Glastonbury, Christine

    2015-01-01

    Invasive facial fungal infections affect the orofacial soft tissues in immunocompromised patients and can cause significant morbidity and mortality. Primary infection occurs from direct inoculation of the skin surface, while secondary infection occurs from extension from an adjacent sinonasal process. The imaging features of secondary infection are similar to acute fulminant invasive fungal sinusitis with infiltration of the orofacial soft tissues in combination with sinonasal disease. Howeve...

  14. Quantitative Evaluation of Tissue Surface Adaption of CAD-Designed and 3D Printed Wax Pattern of Maxillary Complete Denture

    Directory of Open Access Journals (Sweden)

    Hu Chen

    2015-01-01

    Full Text Available Objective. To quantitatively evaluate the tissue surface adaption of a maxillary complete denture wax pattern produced by CAD and 3DP. Methods. A standard edentulous maxilla plaster cast model was used, for which a wax pattern of complete denture was designed using CAD software developed in our previous study and printed using a 3D wax printer, while another wax pattern was manufactured by the traditional manual method. The cast model and the two wax patterns were scanned in the 3D scanner as “DataModel,” “DataWaxRP,” and “DataWaxManual.” After setting each wax pattern on the plaster cast, the whole model was scanned for registration. After registration, the deviations of tissue surface between “DataModel” and “DataWaxRP” and between “DataModel” and “DataWaxManual” were measured. The data was analyzed by paired t-test. Results. For both wax patterns produced by the CAD&RP method and the manual method, scanning data of tissue surface and cast surface showed a good fit in the majority. No statistically significant (P>0.05 difference was observed between the CAD&RP method and the manual method. Conclusions. Wax pattern of maxillary complete denture produced by the CAD&3DP method is comparable with traditional manual method in the adaption to the edentulous cast model.

  15. Rhizobial Inoculation, Alone or Coinoculated with Azospirillum brasilense, Promotes Growth of Wetland Rice

    Directory of Open Access Journals (Sweden)

    Leandro Hahn

    Full Text Available ABSTRACT Rhizobia and associative bacteria promote growth in rice plants (Oryza sativa L. through a series of mechanisms, but most studies on inoculation have been performed based on inoculation with these bacteria in a separate or singular manner. The objective of this study was to assess the efficiency of single/isolated inoculation and inoculation combined with symbiotic rhizobia from forage legume and with Azospirillum brasilense on promoting growth and the root colonization process in wetland rice. Two rhizobia among four isolates from a greenhouse and a laboratory experiment were selected that efficiently promoted seed germination and rice plant growth in a sterilized substrate and in soil. The two most efficient isolates (UFRGS Vp16 and UFRGS Lc348 were inoculated alone or in combination with a commercial product containing A. brasilense in two field experiments using two wetland rice cultivars over two growing seasons. In the field experiments, these isolates coinoculated with A. brasilense promoted larger increases in the agronomic variables of wetland rice compared to the control without inoculation. Confocal laser microscopy confirmed the presence of inoculated bacteria tagged with gfp (UFRGS Vp16, UFRGS Lc348, and A. brasilense colonizing the root surface of the rice seedlings, mainly in the root hairs and lateral roots.

  16. Human isolates of Bartonella tamiae induce pathology in experimentally inoculated immunocompetent mice

    Directory of Open Access Journals (Sweden)

    Kosoy Michael Y

    2010-07-01

    Full Text Available Abstract Background Bartonella tamiae, a newly described bacterial species, was isolated from the blood of three hospitalized patients in Thailand. These patients presented with headache, myalgia, anemia, and mild liver function abnormalities. Since B. tamiae was presumed to be the cause of their illness, these isolates were inoculated into immunocompetent mice to determine their relative pathogenicity in inducing manifestations of disease and pathology similar to that observed in humans. Methods Three groups of four Swiss Webster female mice aged 15-18 months were each inoculated with 106-7 colony forming units of one of three B. tamiae isolates [Th239, Th307, and Th339]. A mouse from each experimental group was sampled at 3, 4, 5 and 6 weeks post-inoculation. Two saline inoculated age-matched controls were included in the study. Samples collected at necropsy were evaluated for the presence of B. tamiae DNA, and tissues were formalin-fixed, stained with hematoxylin and eosin, and examined for histopathology. Results Following inoculation with B. tamiae, mice developed ulcerative skin lesions and subcutaneous masses on the lateral thorax, as well as axillary and inguinal lymphadenopathy. B. tamiae DNA was found in subcutaneous masses, lymph node, and liver of inoculated mice. Histopathological changes were observed in tissues of inoculated mice, and severity of lesions correlated with the isolate inoculated, with the most severe pathology induced by B. tamiae Th239. Mice inoculated with Th239 and Th339 demonstrated myocarditis, lymphadenitis with associated vascular necrosis, and granulomatous hepatitis and nephritis with associated hepatocellular and renal necrosis. Mice inoculated with Th307 developed a deep dermatitis and granulomas within the kidneys. Conclusions The three isolates of B. tamiae evaluated in this study induce disease in immunocompetent Swiss Webster mice up to 6 weeks after inoculation. The human patients from whom these

  17. Early discrimination of nasopharyngeal carcinoma based on tissue deoxyribose nucleic acid surface-enhanced Raman spectroscopy analysis

    Science.gov (United States)

    Qiu, Sufang; Li, Chao; Lin, Jinyong; Xu, Yuanji; Lu, Jun; Huang, Qingting; Zou, Changyan; Chen, Chao; Xiao, Nanyang; Lin, Duo; Chen, Rong; Pan, Jianji; Feng, Shangyuan

    2016-12-01

    Surface-enhanced Raman spectroscopy (SERS) was employed to detect deoxyribose nucleic acid (DNA) variations associated with the development of nasopharyngeal carcinoma (NPC). Significant SERS spectral differences between the DNA extracted from early NPC, advanced NPC, and normal nasopharyngeal tissue specimens were observed at 678, 729, 788, 1337, 1421, 1506, and 1573 cm-1, which reflects the genetic variations in NPC. Principal component analysis combined with discriminant function analysis for early NPC discrimination yielded a diagnostic accuracy of 86.8%, 92.3%, and 87.9% for early NPC, advanced NPC, and normal nasopharyngeal tissue DNA, respectively. In this exploratory study, we demonstrated the potential of SERS for early detection of NPC based on the DNA molecular study of biopsy tissues.

  18. Microtomography evaluation of dental tissue wear surface induced by in vitro simulated chewing cycles on human and composite teeth

    Directory of Open Access Journals (Sweden)

    Rossella Bedini

    2012-01-01

    Full Text Available In this study a 3D microtomography display of tooth surfaces after in vitro dental wear tests has been obtained. Natural teeth have been compared with prosthetic teeth, manufactured by three different polyceramic composite materials. The prosthetic dental element samples, similar to molars, have been placed in opposition to human teeth extracted by paradontology diseases. After microtomography analysis, samples have been subjected to in vitro fatigue test cycles by servo-hydraulic mechanical testing machine. After the fatigue test, each sample has been subjected again to microtomography analysis to obtain volumetric value changes and dental wear surface images. Wear surface images were obtained by 3D reconstruction software and volumetric value changes were measured by CT analyser software. The aim of this work has been to show the potential of microtomography technique to display very clear and reliable wear surface images. Microtomography analysis methods to evaluate volumetric value changes have been used to quantify dental tissue and composite material wear.

  19. [SPREADING OF TISSUE SPHEROIDS FROM PRIMARY HUMAN FIBROBLASTS ON THE SURFACE OF MICROFIBROUS ELECTROSPUN POLYURETHANE MATRIX (A scanning electron microscopic study)].

    Science.gov (United States)

    Kudan, Ye V; Pereira, F D A S; Parfenov, V A; Kasyanov, V A; Khesuani, Yu D; Bulanova, Ye A; Mironov, V A

    2015-01-01

    Tissue spheroids biofabricated from primary human fibroblasts using non-adhesive agarose forms, were placed by 3D bioprinter on the surface of microfibrous electrospun matrix. It was demonstrated that tissue spheroids attached to the surface of matrix during several hours and then gradually spread for several days which indicates high level of biocompatibiity of electrospun microfibrous polyurethane matrix. During this activity, human fibroblasts used processes of leading cell borders for initial step of attachment to matrix filaments. Tissue constructions formed during spreading of tissue spheroids on the surface of electrospun microfibrous polyurethane matrix seem to be a perspective technology platform for development of new methods of biofabrication and 3D bioprinting.

  20. Pulsed erbium laser ablation of hard dental tissue: the effects of atomized water spray versus water surface film

    Science.gov (United States)

    Freiberg, Robert J.; Cozean, Colette D.

    2002-06-01

    It has been established that the ability of erbium lasers to ablate hard dental tissue is due primarily to the laser- initiated subsurface expansion of the interstitial water trapped within the enamel and that by maintaining a thin film of water on the surface of the tooth, the efficiency of the laser ablation is enhanced. It has recently been suggested that a more aggressive ablative mechanism, designated as a hydrokinetic effect, occurs when atomized water droplets, introduced between the erbium laser and the surface of the tooth, are accelerated in the laser's field and impact the tooth's surface. It is the objective of this study to determine if the proposed hydrokinetic effect exists and to establish its contribution to the dental hard tissue ablation process. Two commercially available dental laser systems were employed in the hard tissue ablation studies. One system employed a water irrigation system in which the water was applied directly to the tooth, forming a thin film of water on the tooth's surface. The other system employed pressurized air and water to create an atomized mist of water droplets between the laser hand piece and the tooth. The ablative properties of the two lasers were studied upon hard inorganic materials, which were void of any water content, as well as dental enamel, which contained interstitial water within its crystalline structure. In each case the erbium laser beam was moved across the surface of the target material at a constant velocity. When exposing material void of any water content, no ablation of the surfaces was observed with either laser system. In contrast, when the irrigated dental enamel was exposed to the laser radiation, a linear groove was formed in the enamel surface. The volume of ablated dental tissue associated with each irrigation method was measured and plotted as a function of the energy within the laser pulse. Both dental laser systems exhibited similar enamel ablation rates and comparable ablated surface

  1. Colonization of sorghum and wheat by seed inoculation with Gluconacetobacter diazotrophicus.

    Science.gov (United States)

    Luna, M F; Galar, M L; Aprea, J; Molinari, M L; Boiardi, J L

    2010-08-01

    Colonization of sorghum and wheat after seed inoculation with Gluconacetobacter diazotrophicus strains PAL 5 and UAP 5541/pRGS561 (containing the marker gene gusA) was studied by colony counting and microscopic observation of plant tissues. Inoculum levels as low as 10(2) CFU per seed were enough for root colonization and further spreading in aerial tissues. Rhizoplane colonization was around 7 log CFU g(-1) (fresh weight). G. diazotrophicus was found inside sorghum and wheat roots with populations higher than 5 log CFU g(-1) (fresh weight). Stem colonization remained stable for 30 days post inoculation with endophyte concentrations from 4 to 5 log CFU g(-1) (fresh weight) (in both plants). Population in leaves decreased continuously being undetectable after 17 days post inoculation.

  2. Comparison of Helicobacter pylori colonization on the tonsillar surface versus tonsillar core tissue as determined by the CLO test.

    Science.gov (United States)

    Khademi, Bijan; Niknejad, Nika; Gandomi, Behrooz; Yeganeh, Firoozeh

    2007-08-01

    We conducted a prospective study to determine the correlation between the presence or absence of Helicobacter pylori on the tonsillar surface and in the tonsillar core as determined by the Campylobacter-like organism (CLO) rapid urease enzyme test. Our study population was made up of 55 patients who underwent adenoidectomy, tonsillectomy, or both from December 2002 through April 2003 at Khalili Hospital in Shiraz, Iran. Of these 55 patients, 45 (82%) were positive and 10 (18%) were negative for H pylori colonization as determined by CLO testing. Analysis of samples obtained from individual patients revealed differences in H pylori colonization between tonsillar surface samples and the core tissue samples. Of 106 tonsils obtained from 53 patients who underwent adenotonsillectomy or tonsillectomy, H pylori was found on 56 tonsillar surface samples (53%) and 24 tonsillar core samples (23%); only 13 tonsils (12%) contained H pylori both on the surface and in the core. We conclude that a surface swab is neither specific nor sensitive as an indicator of the presence or absence of H. pylori colonization in tonsillar core tissue.

  3. Conditions Affecting Shelf-Life of Inoculated Legume Seed

    OpenAIRE

    Greg Gemell; Elizabeth Hartley; Rosalind Deaker

    2012-01-01

    Microbial inoculants are becoming more available as sustainable alternatives to fertilizers and other agrichemicals in broad-acre cropping. However, with the exception of legume inoculants little is understood about effective delivery and survival of the inoculum. Legume inoculants are applied to both seed and soil but seed inoculation is the most economical technique. Large quantities of pasture seed in Australia are inoculated by commercial seed coating companies, but the long-term survival...

  4. White-tailed deer are susceptible to the agent of sheep scrapie by intracerebral inoculation.

    Science.gov (United States)

    Greenlee, Justin J; Smith, Jodi D; Kunkle, Robert A

    2011-10-11

    Interspecies transmission studies afford the opportunity to better understand the potential host range and origins of prion diseases. The purpose of this experiment was to determine susceptibility of white-tailed deer to the agent of scrapie after intracerebral inoculation and to compare clinical signs and lesions to those reported for chronic wasting disease (CWD). Deer (n = 5) were inoculated with 1 mL of a 10% (wt/vol) brain homogenate derived from a sheep clinically affected with scrapie. A non-inoculated deer was maintained as a negative control. Deer were observed daily for clinical signs of disease and euthanized and necropsied when unequivocal signs of scrapie were noted. One animal died 7 months post inoculation (pi) due to intercurrent disease. Examinations of brain tissue for the presence of the disease-associated abnormal prion protein (PrP(Sc)) by western blot (WB) and immunohistochemistry (IHC) were negative whereas IHC of lymphoid tissues was positive. Deer necropsied at 15-22 months pi were positive for scrapie by IHC and WB. Deer necropsied after 20 months pi had clinical signs of depression and progressive weight loss. Tissues with PrP(Sc) immunoreactivity included brain (at levels of cerebrum, hippocampus, colliculus, cerebellum, and brainstem), trigeminal ganglion, neurohypophysis, retina, spinal cord, and various lymphoid tissues including tonsil, retropharyngeal and mesenteric lymph nodes, Peyer's patches, and spleen. This work demonstrates for the first time that white-tailed deer are susceptible to sheep scrapie by intracerebral inoculation. To further test the susceptibility of white-tailed deer to scrapie these experiments will be repeated with a more natural route of inoculation.

  5. Expression of transforming growth factor-α and hepatitis B surface antigen in human hepatocellular carcinoma tissues and its significance

    Institute of Scientific and Technical Information of China (English)

    Jing Zhang; Wen-Liang Wang; Qing Li; Qing Qiao

    2004-01-01

    AIM: To evaluate the expression of transforming growth factor-alpha (TGF-α) and hepatitis B surface antigen (HBsAg) in human hepatocellular carcinoma (HCC) tissues and its significance. METHODS: Seventy specimens of HCC tissues were detected by immunohistochemical method. Five specimens of normal human liver tissues were used as control. RESULTS: The TGF-α positive expression rates in HCC and its surrounding tissues were 74.3%(52/70) and 88. t%(52/59), respectively. TGF-α positive granules were mainly in the cytoplasm and fewer existed on the karyotheca. The TGF-α positive expressing rate in well differentiated HCC was significantly higher than that in moderately and poorly differentiated HCC (P<0.05). The TGF-α positive expression also was observed in intrahepatic bile ducts (part of those were hyperplastic ducts). The HBsAg positive expression rates in HCC and its surrounding tissues were 21.4%(15/70) and 79.7%(47/59), respectively. HBsAg positive granules were in the cytoplasm, inclusion and on the karyotheca. There was a prominent positive correlation between TGF-α and HBsAg expression in HCC surrounding tissues (P<0.05, γ=0.34). TGF-α was usually existed with HBsAg in regenerated and/or dysplastic liver cells. In the five normal liver tissues, TGF-α and HBsAg were not detectable in hepatocytes and bile ducts.CONCLUSION: Hepatitis B virus infection is closely related with hepatocarcinogenesis. The overexpression of TGF-α in the liver seems to be associated with the regeneration of hepatocytes injured by HBsAg. The continued expression of TGF-α might lead to dysplasia of liver cells and development of HCC. Furthermore, TGF-α might play a role in morphogenesis and regeneration of intrahepatic bile ducts.

  6. Experimental inoculation of raccoons (Procyon lotor) with Spiroplasma mirum and transmissible mink encephalopathy (TME).

    Science.gov (United States)

    Hamir, Amir N; Greenlee, Justin J; Stanton, Thad B; Smith, Jodi D; Doucette, Stephanie; Kunkle, Robert A; Stasko, Judith A; Richt, Juergen A; Kehrli, Marcus E

    2011-01-01

    The primary objective of this study was to determine whether or not Spiroplasma mirum would be capable of producing lesions of transmissible spongiform encephalopathy (TSE) when inoculated in raccoons (Procyon lotor) and, if that was possible, to compare the clinicopathological findings with those of transmissible mink encephalopathy (TME) in the same experimental model. For this purpose, 5 groups (n = 5) of raccoon kits were inoculated intracerebrally with either S. mirum and/or TME. Two other groups (n = 5) of raccoon kits served as sham-inoculated controls. All animals inoculated with TME, either alone or in combination, showed clinical signs of neurologic disorder and were euthanized within 6 mo post-inoculation (MPI). None of the carcasses revealed gross lesions. Spongiform encephalopathy was observed by light microscopy and the presence of abnormal disease-causing prion protein (PrP(d)) was detected by immunohistochemistry (IHC) and Western blot (WB) techniques in only the raccoons administered TME. Raccoons inoculated with Spiroplasma, but not administered TME agent, were euthanized at 30 MPI. They did not show clinical neurologic signs, their brains did not have lesions of spongiform encephalopathy, and their tissues were negative for S. mirum by polymerase chain reaction (PCR) and for PrP(d) by IHC and WB techniques. The results of this study indicate that Spiroplasma mirum does not induce TSE-like disease in raccoons.

  7. Effect of Arbuscular Mycorrhizal Inoculation on Plant Growth and Phthalic Ester Degradation in Two Contaminated Soils

    Institute of Scientific and Technical Information of China (English)

    CHEN Rui-Rui; YIN Rui; LIN Xian-Gui; CAO Zhi-Hong

    2005-01-01

    A 60-day pot experiment was carried out using di-(2-ethylhexyl) phthalate (DEHP) as a typical organic pollutant phthalic ester and cowpea (Vigna sinensis) as the host plant to determine the effect of arbuscular mycorrhizal inoculation on plant growth and degradation of DEHP in two contaminated soils, a yellow-brown soil and a red soil. The air-dried soils were uniformly sprayed with different concentrations of DEHP, inoculated or left uninoculated with an arbuscular mycorrhizal (AM) fungus, and planted with cowpea seeds. After 60 days the positive impact of AM inoculation on the growth of cowpea was more pronounced in the red soil than in the yellow-brown soil, with significantly higher (P < 0.01)mycorrhizal colonization rate, shoot dry weight and total P content in shoot tissues for the red soil. Both in the yellowbrown and red soils, AM inoculation significantly (P < 0.01) reduced shoot DEHP content, implying that AM inoculation could inhibit the uptake and translocation of DEHP from roots to the aboveground parts. However, with AM inoculation no positive contribution to the degradation of DEHP was found.

  8. Comparison of artificial inoculation methods for studying ...

    African Journals Online (AJOL)

    divya

    2013-05-01

    May 1, 2013 ... Establishment of disease by artificial inoculation is essential for studies of various aspects of plant pathology. Keeping this in mind .... three to four days under cool white fluorescent light (2,000 lux) followed by 2 days ... and placed in growth chambers at 25°C and 70% relative humidity. The Petri plates were ...

  9. Coping With Pain: Studies in Stress Inoculation.

    Science.gov (United States)

    Horan, John J.; And Others

    The stress-inoculation paradigm for helping clients deal with pain consists of education about the psychological dimensions of pain, training in a number of coping skills relevant to each dimension, and practice in applying these skills to the noxious stimulus. Presented are two studies, the first of which represents a component analysis of stress…

  10. Graminan breakdown by fructan exohydrolase induced in winter wheat inoculated with snow mold.

    Science.gov (United States)

    Kawakami, Akira; Yoshida, Midori

    2012-02-15

    Fructan structures vary widely among plant species. Graminan-type fructans, extensions of sucrose through β-(2,6)-linked fructosyl units with branches of β-(2,1)-linked fructosyl units, accumulate in tissues of winter wheat (Triticum aestivum) during cold hardening and are metabolized under persistent snow cover. Snow molds such as Typhula ishikariensis and Microdochium nivale opportunistically infect wheat under snow cover. Snow mold-resistant wheat cultivars tend to heavily accumulate and slowly metabolize water-soluble carbohydrates including graminans in comparison with snow mold-susceptible cultivars. We observed time-dependent changes in the amounts of water-soluble carbohydrates in snow mold-inoculated wheat tissues, and accumulated fructan levels significantly decreased as a result of snow mold inoculation and incubation under snow cover, especially in a snow mold-susceptible wheat cultivar. Three candidates for fructan exohydrolase (FEH) cDNAs with high homology to cell wall invertases were isolated from wheat leaf tissues inoculated with snow mold and incubated under snow cover. The substrate specificity of enzymes encoded by the isolated clones was analyzed by recombinant proteins expressed in Pichia pastoris. The recombinant protein (Wfh-sm3m) encoded by one (Wfh-sm3) of the isolated clones preferentially degraded 6-kestotriose and possessed minor hydrolase activity to 1-kestotriose and 1,1-kestotetraose. Moreover, Wfh-sm3m hydrolyzed almost all graminans that accumulated in hardened wheat tissues. Wfh-sm3 transcripts increased in wheat leaf tissues inoculated with snow mold and incubated under snow cover. These results suggest that Wfh-sm3 encodes a 6-FEH with minor 1-FEH activity and is associated with degradation of fructans in wheat leaf tissues during inoculation and incubation under snow cover.

  11. Colonization and beneficial effects on annual ryegrass by mixed inoculation with plant growth promoting bacteria.

    Science.gov (United States)

    Castanheira, Nádia L; Dourado, Ana Catarina; Pais, Isabel; Semedo, José; Scotti-Campos, Paula; Borges, Nuno; Carvalho, Gilda; Barreto Crespo, Maria Teresa; Fareleira, Paula

    2017-05-01

    Multi-strain inoculants have increased potential to accomplish a diversity of plant needs, mainly attributed to its multi-functionality. This work evaluated the ability of a mixture of three bacteria to colonize and induce a beneficial response on the pasture crop annual ryegrass. Pseudomonas G1Dc10 and Paenibacillus G3Ac9 were previously isolated from annual ryegrass and were selected for their ability to perform multiple functions related to plant growth promotion. Sphingomonas azotifigens DSMZ 18530(T) was included due to nitrogen fixing ability. The effects of the bacterial mixture were assessed in gnotobiotic plant inoculation assays and compared with single and dual inoculation treatments. Triple inoculation with 3×10(8) bacteria significantly increased plant dry weight and leaf pigments, indicating improved photosynthetic performance. Plant lipid biosynthesis was enhanced by 65%, mainly due to the rise of linolenic acid, an omega-3 fatty acid with high dietary value. Electrolyte leakage, an indicator of plant membrane stability under stress, was decreased pointing to a beneficial effect by inoculation. Plants physiological condition was more favoured by triple inoculation than by single, although benefits on biomass were only evident relative to non-inoculated plants. The colonization behaviour and coexistence in plant tissues were assessed using FISH and GFP-labelling, combined with confocal microscopy and a cultivation-based approach for quantification. The three strains occupied the same sites, localizing preferentially along root hairs and in stem epidermis. Endophytic colonization was observed as bacteria entered root and stem inner tissues. This study reveals the potential of this mixture of strains for biofertilization, contributing to improve crop productivity and nutritional value. Copyright © 2017 Elsevier GmbH. All rights reserved.

  12. Tobacco Mosaic Virus in the Lungs of Mice following Intra-Tracheal Inoculation

    Science.gov (United States)

    Balique, Fanny; Colson, Philippe; Barry, Abdoulaye Oury; Nappez, Claude; Ferretti, Audrey; Moussawi, Khatoun Al; Ngounga, Tatsiana; Lepidi, Hubert; Ghigo, Eric; Mege, Jean-Louis; Lecoq, Hervé; Raoult, Didier

    2013-01-01

    Plant viruses are generally considered incapable of infecting vertebrates. Accordingly, they are not considered harmful for humans. However, a few studies questioned the certainty of this paradigm. Tobacco mosaic virus (TMV) RNA has been detected in human samples and TMV RNA translation has been described in animal cells. We sought to determine if TMV is detectable, persists, and remains viable in the lung tissues of mice following intratracheal inoculation, and we attempted to inoculate mouse macrophages with TMV. In the animal model, mice were intratracheally inoculated with 1011 viral particles and were sacrificed at different time points. The virus was detected in the mouse lungs using immunohistochemistry, electron microscopy, real-time RT-PCR and sequencing, and its viability was studied with an infectivity assay on plants. In the cellular model, the culture medium of murine bone marrow derived macrophages (BMDM) was inoculated with different concentrations of TMV, and the virus was detected with real-time RT-PCR and immunofluorescence. In addition, anti-TMV antibodies were detected in mouse sera with ELISA. We showed that infectious TMV could enter and persist in mouse lungs via the intratracheal route. Over 14 days, the TMV RNA level decreased by 5 log10 copies/ml in the mouse lungs and by 3.5 log10 in macrophages recovered from bronchoalveolar lavage. TMV was localized to lung tissue, and its infectivity was observed on plants until 3 days after inoculation. In addition, anti-TMV antibody seroconversions were observed in the sera from mice 7 days after inoculation. In the cellular model, we observed that TMV persisted over 15 days after inoculation and it was visualized in the cytoplasm of the BMDM. This work shows that a plant virus, Tobacco mosaic virus, could persist and enter in cells in mammals, which raises questions about the potential interactions between TMV and human hosts. PMID:23383021

  13. Implications of Surface and Bulk Properties of Abutment Implants and Their Degradation in the Health of Periodontal Tissue

    Directory of Open Access Journals (Sweden)

    Erica Dorigatti de Avila

    2013-12-01

    Full Text Available The aim of the current review was to investigate the implications of the surface and bulk properties of abutment implants and their degradation in relation to periodontal health. The success of dental implants is no longer a challenge for dentistry. The scientific literature presents several types of implants that are specific for each case. However, in cases of prosthetics components, such as abutments, further research is needed to improve the materials used to avoid bacterial adhesion and enhance contact with epithelial cells. The implanted surfaces of the abutments are composed of chemical elements that may degrade under different temperatures or be damaged by the forces applied onto them. This study showed that the resulting release of such chemical elements could cause inflammation in the periodontal tissue. At the same time, the surface characteristics can be altered, thus favoring biofilm development and further increasing the inflammation. Finally, if not treated, this inflammation can cause the loss of the implant.

  14. Multi-dimensional TOF-SIMS analysis for effective profiling of disease-related ions from the tissue surface

    Science.gov (United States)

    Park, Ji-Won; Jeong, Hyobin; Kang, Byeongsoo; Kim, Su Jin; Park, Sang Yoon; Kang, Sokbom; Kim, Hark Kyun; Choi, Joon Sig; Hwang, Daehee; Lee, Tae Geol

    2015-06-01

    Time-of-flight secondary ion mass spectrometry (TOF-SIMS) emerges as a promising tool to identify the ions (small molecules) indicative of disease states from the surface of patient tissues. In TOF-SIMS analysis, an enhanced ionization of surface molecules is critical to increase the number of detected ions. Several methods have been developed to enhance ionization capability. However, how these methods improve identification of disease-related ions has not been systematically explored. Here, we present a multi-dimensional SIMS (MD-SIMS) that combines conventional TOF-SIMS and metal-assisted SIMS (MetA-SIMS). Using this approach, we analyzed cancer and adjacent normal tissues first by TOF-SIMS and subsequently by MetA-SIMS. In total, TOF- and MetA-SIMS detected 632 and 959 ions, respectively. Among them, 426 were commonly detected by both methods, while 206 and 533 were detected uniquely by TOF- and MetA-SIMS, respectively. Of the 426 commonly detected ions, 250 increased in their intensities by MetA-SIMS, whereas 176 decreased. The integrated analysis of the ions detected by the two methods resulted in an increased number of discriminatory ions leading to an enhanced separation between cancer and normal tissues. Therefore, the results show that MD-SIMS can be a useful approach to provide a comprehensive list of discriminatory ions indicative of disease states.

  15. The Application of Minimally Invasive Devices with Nanostructured Surface Functionalization: Antisticking Behavior on Devices and Liver Tissue Interface in Rat

    Directory of Open Access Journals (Sweden)

    Li-Hsiang Lin

    2015-01-01

    Full Text Available This study investigated the thermal injury and adhesion property of a novel electrosurgery of liver using copper-doped diamond-like carbon (DLC-Cu surface treatment. It is necessary to reduce the thermal damage of surrounding tissues for clinical electrosurgeries. The surface morphologies of stainless steel (SS coated with DLC (DLC-Cu-SS films were characterized by scanning electron microscopy (SEM and transmission electron microscopy (TEM. Bionic liver models were reconstructed using magnetic resonance imaging (MRI to simulate electrosurgery. Cell cytotoxicity assays showed that the DLC-Cu thin film was nontoxic. The temperature of tissue decreased significantly with use of the electrosurgical device with nanostructured DLC-Cu films and increased with increasing thickness of the films. Thermography revealed that the surgical temperature in the DLC-Cu-SS electrosurgical device was significantly lower than that in the untreated device in the animal model. Moreover, compared to the SS electrosurgical device, the DLC-Cu-SS electrosurgical device caused a relatively small injury area and lateral thermal effect. The results indicate that the DLC-Cu-SS electrosurgical device decreases excessive thermal injury and ensures homogeneous temperature transformation in the tissues.

  16. The effect of disinfectant agents on dimensional stability and surface roughness of a tissue conditioner material

    Directory of Open Access Journals (Sweden)

    Amiralireza Khaledi

    2011-01-01

    Conclusion: The results showed that the disinfectant solutions used in this study did not have a significant effect on the surface quality and dimensional accuracy of Visco-gel as a functional impression material.

  17. Conservation of somatic tissue derived from collared peccaries (Pecari tajacu Linnaeus, 1758) using direct or solid-surface vitrification techniques.

    Science.gov (United States)

    Borges, Alana Azevedo; Lima, Gabriela Liberalino; de Queiroz Neta, Luiza Bento; Santos, Maria Valéria de Oliveira; de Oliveira, Moacir Franco; Silva, Alexandre Rodrigues; Pereira, Alexsandra Fernandes

    2017-08-01

    Cryopreservation of somatic tissue can be applied in biodiversity conservation, especially for wild species as collared peccary. We aimed to evaluate the effect of vitrification techniques of ear tissue of collared peccary [direct vitrification in cryovials (DVC) or solid-surface vitrification (SSV)] on the layers of epidermis and dermis by conventional histology and cell ability during the in vitro culture. Thus, both the vitrification methods were able to maintain normal patterns of the epidermis as the cornea and granular layers, furthermore the intercellular space and dermal-epidermal junction of the spinous layer when compared to fresh control. Nevertheless, DVC and SSV percentage of normality decreased in the morphological integrity of cytoplasm (37.5 and 25.0%) of spinous layer, respectively, as compared to the fresh fragments (100%, p  0.05). In addition, only at time of 72 h (D3), in the growth curve, DVC fragments showed a reduced cell concentration than fresh control. In conclusion, SSV was found to be a more efficient method for vitrifying collared peccary skin tissue when compared to DVC. These results are relevant for the tissue cryopreservation from collared peccary and could also be useful for mammals with phylogenetic relationships.

  18. Directing and Potentiating Stem Cell-Mediated Angiogenesis and Tissue Repair by Cell Surface E-Selectin Coating.

    Science.gov (United States)

    Liu, Zhao-Jun; Daftarian, Pirouz; Kovalski, Letícia; Wang, Bo; Tian, Runxia; Castilla, Diego M; Dikici, Emre; Perez, Victor L; Deo, Sapna; Daunert, Sylvia; Velazquez, Omaida C

    2016-01-01

    Stem cell therapy has emerged as a promising approach for treatment of a number of diseases, including delayed and non-healing wounds. However, targeted systemic delivery of therapeutic cells to the dysfunctional tissues remains one formidable challenge. Herein, we present a targeted nanocarrier-mediated cell delivery method by coating the surface of the cell to be delivered with dendrimer nanocarriers modified with adhesion molecules. Infused nanocarrier-coated cells reach to destination via recognition and association with the counterpart adhesion molecules highly or selectively expressed on the activated endothelium in diseased tissues. Once anchored on the activated endothelium, nanocarriers-coated transporting cells undergo transendothelial migration, extravasation and homing to the targeted tissues to execute their therapeutic role. We now demonstrate feasibility, efficacy and safety of our targeted nanocarrier for delivery of bone marrow cells (BMC) to cutaneous wound tissues and grafted corneas and its advantages over conventional BMC transplantation in mouse models for wound healing and neovascularization. This versatile platform is suited for targeted systemic delivery of virtually any type of therapeutic cell.

  19. Real-Time Visualization of Tissue Surface Biochemical Features Derived From Fluorescence Lifetime Measurements.

    Science.gov (United States)

    Gorpas, Dimitris; Ma, Dinglong; Bec, Julien; Yankelevich, Diego R; Marcu, Laura

    2016-08-01

    Fiber based fluorescence lifetime imaging has shown great potential for intraoperative diagnosis and guidance of surgical procedures. Here we describe a novel method addressing a significant challenge for the practical implementation of this technique, i.e., the real-time display of the quantified biochemical or functional tissue properties superimposed on the interrogated area. Specifically, an aiming beam (450 nm) generated by a continuous-wave laser beam was merged with the pulsed fluorescence excitation light in a single delivery/collection fiber and then imaged and segmented using a color-based algorithm. We demonstrate that this approach enables continuous delineation of the interrogated location and dynamic augmentation of the acquired frames with the corresponding fluorescence decay parameters. The method was evaluated on a fluorescence phantom and fresh tissue samples. Current results demonstrate that 34 frames per second can be achieved for augmenting videos of 640 × 512 pixels resolution. Also we show that the spatial resolution of the fluorescence lifetime map depends on the tissue optical properties, the scanning speed, and the frame rate. The dice similarity coefficient between the fluorescence phantom and the reconstructed maps was estimated to be as high as 93%. The reported method could become a valuable tool for augmenting the surgeon's field of view with diagnostic information derived from the analysis of fluorescence lifetime data in real-time using handheld, automated, or endoscopic scanning systems. Current method provides also a means for maintaining the tissue light exposure within safety limits. This study provides a framework for using an aiming beam with other point spectroscopy applications.

  20. Surface modification of copolymerized films from three-armed biodegradable macromers - An analytical platform for modified tissue engineering scaffolds.

    Science.gov (United States)

    Müller, Benno M; Loth, Rudi; Hoffmeister, Peter-Georg; Zühl, Friederike; Kalbitzer, Liv; Hacker, Michael C; Schulz-Siegmund, Michaela

    2017-03-15

    The concept of macromers allows for a broad adjustment of biomaterial properties by macromer chemistry or copolymerization. Copolymerization strategies can also be used to introduce reactive sites for subsequent surface modification. Control over surface features enables adjustment of cellular reactions with regard to site and object of implantation. We designed macromer-derived polymer films which function as non-implantable analytical substrates for the investigation of surface properties of equally composed scaffolds for bone tissue engineering. To this end, a toolbox of nine different biodegradable, three-armed macromers was thermally cross-copolymerized with poly(ethylene glycol)-methacrylate (PEG-MA) to films. Subsequent activation of PEG-hydroxyl groups with succinic anhydride and N-hydroxysuccinimid allowed for covalent surface modification. We quantified the capacity to immobilize analytes of low (amino-functionalized fluorescent dye, Fcad, and RGD-peptides) and high (alkaline phosphatase, ALP) molecular weight. Fcad grafting level was controlled by macromer chemistry, content and molecular weight of PEG-MA, but also the solvent used for film synthesis. Fcad molar amount per surface area was twentyfive times higher on high-swelling compared to low-swelling films, but differences became smaller when large ALP (appr. 2:1) were employed. Similarly, small differences were observed on RGD peptide functionalized films that were investigated by cell adhesion studies. Presentation of PEG-derivatives on surfaces was visualized by atomic force microscopy (AFM) which unraveled composition-dependent domain formation influencing fluorescent dye immobilization. Surface wetting characteristics were investigated via static water contact angle. We conclude that macromer ethoxylation and lactic acid content determined film swelling, PEG domain formation and eventually efficiency of surface decoration.

  1. [Analysis of Proliferation and Survival of Agrobacteria after Inoculation of Maize Pistil Filaments].

    Science.gov (United States)

    Volokhina, V; Velikov, V A; Moiseeva, E M; Chumakov, M I

    2016-01-01

    While the authors have previously developed a method of pistil filament treatment with Agrobacterium cells during blossoming for the transformation of maize generative cells, the mechanism for bacterial T-DNA penetration into the embryo sac remained unknown. This article analyzes the possibility of agrobacterial penetration into the maize embryo via pollen tubes. Microbiological, PCR, and GUS techniques were used to confirm that agrobacteria could spread for up to 20 cm from the sie of inoculation and were detected in maize embryo tissues as aerly as 24 h after inoculation, while they were not revealed after 5-13 days.

  2. Decompression induced bubble dynamics on ex vivo fat and muscle tissue surfaces with a new experimental set up.

    Science.gov (United States)

    Papadopoulou, Virginie; Evgenidis, Sotiris; Eckersley, Robert J; Mesimeris, Thodoris; Balestra, Costantino; Kostoglou, Margaritis; Tang, Meng-Xing; Karapantsios, Thodoris D

    2015-05-01

    Vascular gas bubbles are routinely observed after scuba dives using ultrasound imaging, however the precise formation mechanism and site of these bubbles are still debated and growth from decompression in vivo has not been extensively studied, due in part to imaging difficulties. An experimental set-up was developed for optical recording of bubble growth and density on tissue surface area during hyperbaric decompression. Muscle and fat tissues (rabbits, ex vivo) were covered with nitrogen saturated distilled water and decompression experiments performed, from 3 to 0bar, at a rate of 1bar/min. Pictures were automatically acquired every 5s from the start of the decompression for 1h with a resolution of 1.75μm. A custom MatLab analysis code implementing a circular Hough transform was written and shown to be able to track bubble growth sequences including bubble center, radius, contact line and contact angles over time. Bubble density, nucleation threshold and detachment size, as well as coalescence behavior, were shown significantly different for muscle and fat tissues surfaces, whereas growth rates after a critical size were governed by diffusion as expected. Heterogeneous nucleation was observed from preferential sites on the tissue substrate, where the bubbles grow, detach and new bubbles form in turn. No new nucleation sites were observed after the first 10min post decompression start so bubble density did not vary after this point in the experiment. In addition, a competition for dissolved gas between adjacent multiple bubbles was demonstrated in increased delay times as well as slower growth rates for non-isolated bubbles.

  3. Treating Cutaneous T-cell Lymphoma with Highly Irregular Surfaces with Photon Irradiation Using Rice as Tissue Compensator

    Directory of Open Access Journals (Sweden)

    Lonika eMajithia

    2015-02-01

    Full Text Available Purpose: Cutaneous T-cell lymphoma (CTCL is known to have an excellent response to radiotherapy, an important treatment modality for this disease. In patients with extremity and digit involvement, the irregular surface and depth variations create difficulty in delivering a homogenous dose using electrons. We sought to evaluate photon irradiation with rice packing as tissue equivalence and determine clinical tolerance and response. Materials and Methods: Three consecutive CTCL patients with extensive lower extremity involvement including the digits were treated using external beam photon therapy with rice packing for tissue compensation. The entire foot was treated to 30-40 Gy in 2-3 Gy per fraction using 6 MV photons prescribed to the mid-plane of an indexed box filled with rice in which the foot was placed. Optically stimulated luminescence dosimeter (OSLD was used for dose measurement to determine the dose deposition to the skin surface. Treatment tolerance and response were monitored with clinical evaluation. Results: All patients tolerated the treatment without treatment breaks. Toxicities included grade 3 erythema and desquamation with resolution within 4 weeks. No late toxicities were observed. All four treated sites had partial response (PR by the end of the treatment course. All patients reported improved functionality after treatment, with less pain, drainage, or swelling. No local recurrence has been observed in these patients with a median follow-up time of 14 months. Conclusion: Tissue compensation with rice packing offers a convenient, inexpensive and reproducible method for the treatment of CTCL with highly irregular surfaces.

  4. Surface modification of nanofibrous polycaprolactone/gelatin composite scaffold by collagen type I grafting for skin tissue engineering

    Energy Technology Data Exchange (ETDEWEB)

    Gautam, Sneh [Department of Polymer and Process Engineering, Indian Institute of Technology Roorkee (India); Chou, Chia-Fu [Institute of Physics, Academia Sinica, Taipei, Taiwan (China); Dinda, Amit K. [Department of Pathology, All India Institute of Medical Science, New Delhi (India); Potdar, Pravin D. [Department of Molecular Medicine and Biology, Jaslok Hospital and Research Centre, Mumbai (India); Mishra, Narayan C., E-mail: mishrawise@gmail.com [Department of Polymer and Process Engineering, Indian Institute of Technology Roorkee (India)

    2014-01-01

    In the present study, a tri-polymer polycaprolactone (PCL)/gelatin/collagen type I composite nanofibrous scaffold has been fabricated by electrospinning for skin tissue engineering and wound healing applications. Firstly, PCL/gelatin nanofibrous scaffold was fabricated by electrospinning using a low cost solvent mixture [chloroform/methanol for PCL and acetic acid (80% v/v) for gelatin], and then the nanofibrous PCL/gelatin scaffold was modified by collagen type I (0.2–1.5 wt.%) grafting. Morphology of the collagen type I-modified PCL/gelatin composite scaffold that was analyzed by field emission scanning electron microscopy (FE-SEM), showed that the fiber diameter was increased and pore size was decreased by increasing the concentration of collagen type I. Fourier transform infrared (FT-IR) spectroscopy and thermogravimetric (TG) analysis indicated the surface modification of PCL/gelatin scaffold by collagen type I immobilization on the surface of the scaffold. MTT assay demonstrated the viability and high proliferation rate of L929 mouse fibroblast cells on the collagen type I-modified composite scaffold. FE-SEM analysis of cell-scaffold construct illustrated the cell adhesion of L929 mouse fibroblasts on the surface of scaffold. Characteristic cell morphology of L929 was also observed on the nanofiber mesh of the collagen type I-modified scaffold. Above results suggest that the collagen type I-modified PCL/gelatin scaffold was successful in maintaining characteristic shape of fibroblasts, besides good cell proliferation. Therefore, the fibroblast seeded PCL/gelatin/collagen type I composite nanofibrous scaffold might be a potential candidate for wound healing and skin tissue engineering applications. - Highlights: • PCL/gelatin/collagen type I scaffold was fabricated for skin tissue engineering. • PCL/gelatin/collagen type I scaffold showed higher fibroblast growth than PCL/gelatin one. • PCL/gelatin/collagen type I might be one of the ideal scaffold for

  5. Inoculation Stress Hypothesis of Environmental Enrichment

    Science.gov (United States)

    Crofton, Elizabeth J.; Zhang, Yafang; Green, Thomas A.

    2014-01-01

    One hallmark of psychiatric conditions is the vast continuum of individual differences in susceptibility vs. resilience resulting from the interaction of genetic and environmental factors. The environmental enrichment paradigm is an animal model that is useful for studying a range of psychiatric conditions, including protective phenotypes in addiction and depression models. The major question is how environmental enrichment, a non-drug and non-surgical manipulation, can produce such robust individual differences in such a wide range of behaviors. This paper draws from a variety of published sources to outline a coherent hypothesis of inoculation stress as a factor producing the protective enrichment phenotypes. The basic tenet suggests that chronic mild stress from living in a complex environment and interacting non-aggressively with conspecifics can inoculate enriched rats against subsequent stressors and/or drugs of abuse. This paper reviews the enrichment phenotypes, mulls the fundamental nature of environmental enrichment vs. isolation, discusses the most appropriate control for environmental enrichment, and challenges the idea that cortisol/corticosterone equals stress. The intent of the inoculation stress hypothesis of environmental enrichment is to provide a scaffold with which to build testable hypotheses for the elucidation of the molecular mechanisms underlying these protective phenotypes and thus provide new therapeutic targets to treat psychiatric/neurological conditions. PMID:25449533

  6. Concordance of human papillomavirus types detected on the surface and in the tissue of genital lesions in men

    OpenAIRE

    Anic, Gabriella M; Messina, Jane L.; Stoler, Mark H.; Rollison, Dana E.; Stockwell, Heather; Villa, Luisa L.; Lazcano-Ponce, Eduardo; Gage, Christine; Silva, Roberto Jose C.; Baggio, Maria L.; Salmerón, Jorge; Giuliano, Anna R.

    2013-01-01

    Swabbing the surface of a genital lesion to obtain a sample for HPV DNA testing is less invasive than a biopsy, but may not represent HPV types present in the lesion tissue. The objective of this study was to examine the concordance of HPV types detected in swab and biopsy samples from 165 genital lesions from men ages 18-70. Lesions included 90 condyloma, 10 penile intraepithelial neoplasia (PeIN), 23 non-condyloma with a known histology, and 42 lesions with an undetermined histology. All le...

  7. Optical multi-frequency swept sensing for wide-field vibration measurement of interior surfaces in biological tissue

    Science.gov (United States)

    Choi, S.; Nin, F.; Hibino, H.; Suzuki, T.

    2015-12-01

    Multifrequency sensing technique adopting the wide field heterodyne detection technique is demonstrated for interior surface vibration measurements in thick biological tissue. These arrangements allow obtaining not only 3D tomographic images but also various vibration parameters such as spatial amplitude, phase, and frequency, with high temporal and transverse resolutions over a wide field. The axial resolution and the accuracy of vibration amplitude measurement were estimated to be 2.5 μm and 3 nm, respectively. This wide-field tomographic sensing method can be applied for measuring microdynamics of a variety of biological samples, thus contributing to the progress in life sciences research.

  8. FEASIBILITY STUDY OF BUSINESS IN AGARWOOD INOCULATION AT DIFFERENT STEM DIAMETERS AND INOCULATION PERIODS

    Directory of Open Access Journals (Sweden)

    Sri Suharti

    2011-12-01

    Full Text Available Indonesia signifies as the biggest agarwood producer country in the world. Its demand and price tend to increase and have brought about over exploitation of agarwood. Consequently, its population in nature has decreased significantly. To overcome the situation, since 1995, agarwood has been included in the CITES Appendix II. However, illegal exploitation remains persistent and reaches an excessive level. In order to deal with it, agarwood cultivation and its artificial production have been undertaken at several provinces in Indonesia. Some supporting factors for agarwood cultivation and artificial production are the availability of potential land for extensive cultivation, appropriate agro climate condition, simple cultivation technique and already being well adopted by farmers, the availability of necessary pathogen for agarwood inoculation, and the increasing demand with relatively high price. The research aims to analyze the feasibility study of agarwood inoculation business at several stem diameters (15 - 25 cm; 26 -35 cm and 36 - 40 cm and periods of inoculation (1 - 5 years. Data were collected through field observation and literature study. The results showed that inoculation on agarwood producer tree stands at 12.5% interest rate afforded positive net present value (NPV,  internal rate of return (IRR is much higher than market interest and benefit cost (B/C ratio >2 for those three diameter classes. Furthermore, if agarwood harvesting is delayed until five years after inoculation, NPV,  IRR and B/C ratio would be much higher. It can be concluded that inoculation on agarwood producer tree stands (at appropriate age for inoculation is feasible to be developed.

  9. Extracellular matrix and cell surface as determinants of connective tissue differentiation.

    Science.gov (United States)

    Solursh, M

    1989-09-01

    This paper reviews in vitro studies, largely from the author's laboratory, concerning the conditions that are permissive for the differentiation of limb bud mesenchymal cells into chondrocytes. In high-density cell culture, even in a defined medium, the same normal sequence of events that is found in vivo in developing cartilage is also observed. This system can be used to study heritable disorders in model systems such as in mutant mouse embryos. In addition, single mesenchymal cells can differentiate into hypertrophic chondrocytes in hydrated collagen gel or agarose cultures. A rounded cell shape promotes chondrogenesis, while a flattened cell shape promotes fibroblast differentiation. The actin cytoskeleton is shown to play a central role in regulating connective tissue cell differentiation. By use of such cell culture manipulations, it is now possible to grow large numbers of fibroblastic cells from human biopsy material for storage and to carry out experimental studies after re-expression of chondrogenesis in gel cultures. It is suggested that cytoskeletal-extracellular matrix interactions play a fundamental role in connective tissue differentiation. Matrix receptors might be developmentally regulated and modify epithelial effects on mesenchymal cells. In this way mesenchymal cells differentiate in a highly organized manner in spatial and temporal terms.

  10. Theory and experiment for the effect of vascular microstructure on surface tissue heat transfer--Part I: Anatomical foundation and model conceptualization.

    Science.gov (United States)

    Weinbaum, S; Jiji, L M; Lemons, D E

    1984-11-01

    A new theoretical model supported by ultrastructural studies and high-spatial resolution temperature measurements is presented for surface tissue heat transfer in a two-part study. In this first paper, vascular casts of the rabbit thigh prepared by the tissue clearance method were serially sectioned parallel to the skin surface to determine the detailed variation of the vascular geometry as a function of tissue depth. Simple quantitative models of the basic vascular structures observed were then analyzed in terms of their characteristic thermal relaxation lengths and a new three-layer conceptual model proposed for surface tissue heat transfer. Fine wire temperature measurements with an 80-micron average diameter thermocouple junction and spatial increments of 20 micrometers between measurement sites have shown for the first time the detailed temperature fluctuations in the microvasculature and have confirmed the fundamental assumptions of the proposed three-layer model for the deep tissue, skeletal muscle and cutaneous layers.

  11. Review of Microbial Inoculation Agent Used in Composting

    Institute of Scientific and Technical Information of China (English)

    HUANG Yaxi; FENG Shuqing; XU Xiuhong; MAO Zhicheng; WANG Mingyue; LIU Yanfang; HUANG Yan

    2009-01-01

    Microbial inoculation agent has been widely used in waste disposal and waste recycling. In this paper, the principle of fermentation in the composting of agricultural waste and the impact of microbial inoculation agent on composting were reviewed.The research and the application of microbial inoculation agent were also introduced. Based on the trend of the research, the promising prospect of microbial inoculation agent was discussed.

  12. Accessing inoculation methods of maize and wheat with Azospirillum brasilense

    OpenAIRE

    FUKAMI,J.; M. A. Nogueira; ARAUJO, R. S.; Hungria, M.

    2016-01-01

    The utilization of inoculants containing Azospirillum is becoming more popular due to increasing reports of expressive gains in grain yields. However, incompatibility with pesticides used in seed treatments represents a main limitation for a successful inoculation. Therefore, in this study we searched for alternatives methods for seed inoculation of maize and wheat, aiming to avoid the direct contact of bacteria with pesticides. Different doses of inoculants containing Azospirillum brasilense...

  13. Histologic studies on osseointegration: soft tissues response to implant surfaces and components. A review.

    Science.gov (United States)

    Piattelli, Adriano; Pontes, Ana Emilia Farias; Degidi, Marco; Iezzi, Giovanna

    2011-01-01

    It is important to clarify the potential response of different types of cells to different implant materials and topographies. Thus, in vitro studies are performed using cell cultures, in order to evaluate, among other characteristics, the morphology, orientation, proliferation and adhesion of the cells. Histology evaluation are performed in animals or humans to describe the physiological response to different surfaces. Copyright © 2010 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.

  14. 7 CFR 201.24a - Inoculated seed.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 3 2010-01-01 2010-01-01 false Inoculated seed. 201.24a Section 201.24a Agriculture..., Inspections, Marketing Practices), DEPARTMENT OF AGRICULTURE (CONTINUED) FEDERAL SEED ACT FEDERAL SEED ACT REGULATIONS Labeling Agricultural Seeds § 201.24a Inoculated seed. Seed claimed to be inoculated shall...

  15. A Solvent-Free Surface Suspension Melt Technique for Making Biodegradable PCL Membrane Scaffolds for Tissue Engineering Applications.

    Science.gov (United States)

    Suntornnond, Ratima; An, Jia; Tijore, Ajay; Leong, Kah Fai; Chua, Chee Kai; Tan, Lay Poh

    2016-03-21

    In tissue engineering, there is limited availability of a simple, fast and solvent-free process for fabricating micro-porous thin membrane scaffolds. This paper presents the first report of a novel surface suspension melt technique to fabricate a micro-porous thin membrane scaffolds without using any organic solvent. Briefly, a layer of polycaprolactone (PCL) particles is directly spread on top of water in the form of a suspension. After that, with the use of heat, the powder layer is transformed into a melted layer, and following cooling, a thin membrane is obtained. Two different sizes of PCL powder particles (100 µm and 500 µm) are used. Results show that membranes made from 100 µm powders have lower thickness, smaller pore size, smoother surface, higher value of stiffness but lower ultimate tensile load compared to membranes made from 500 µm powder. C2C12 cell culture results indicate that the membrane supports cell growth and differentiation. Thus, this novel membrane generation method holds great promise for tissue engineering.

  16. Is Graphene a Promising Nano-Material for Promoting Surface Modification of Implants or Scaffold Materials in Bone Tissue Engineering?

    Science.gov (United States)

    Gu, Ming; Liu, Yunsong; Chen, Tong; Du, Feng; Zhao, Xianghui; Xiong, Chunyang

    2014-01-01

    Bone tissue engineering promises to restore bone defects that are caused by severe trauma, congenital malformations, tumors, and nonunion fractures. How to effectively promote the proliferation and osteogenic differentiation of mesenchymal stem cells (MSCs) or seed cells has become a hot topic in this field. Many researchers are studying the ways of conferring a pro-osteodifferentiation or osteoinductive capability on implants or scaffold materials, where osteogenesis of seed cells is promoted. Graphene (G) provides a new kind of coating material that may confer the pro-osteodifferentiation capability on implants and scaffold materials by surface modification. Here, we review recent studies on the effects of graphene on surface modifications of implants or scaffold materials. The ability of graphene to improve the mechanical and biological properties of implants or scaffold materials, such as nitinol and carbon nanotubes, and its ability to promote the adhesion, proliferation, and osteogenic differentiation of MSCs or osteoblasts have been demonstrated in several studies. Most previous studies were performed in vitro, but further studies will explore the mechanisms of graphene's effects on bone regeneration, its in vivo biocompatibility, its ability to promote osteodifferentiation, and its potential applications in bone tissue engineering. PMID:24447041

  17. The dynamics of apoplast phenolics in tobacco leaves following inoculation with bacteria

    Directory of Open Access Journals (Sweden)

    Con Jacyn Baker

    2015-08-01

    Full Text Available This study demonstrates that the accumulation of apoplastic phenolics is stimulated in planta in response to bacterial inoculation. Past studies have shown that levels of extracellular phenolics are elicited in plant cell suspensions in response to bacteria, and that tomato plants infected with viroids showed changes in apoplastic phenolics. The method described here monitored changes in apoplastic phenolics in tobacco leaves following bacterial inoculation of the same tissue. Inoculation with a saprophyte, Pseudomonas fluorescens, which does not cause visible symptoms or physical damage, was used to elicit phenolics and examine the effects of variable parameters on phenolic composition. Location of the inoculation on the leaf, position or developmental age of the leaf on the plant, and inoculum concentration were standardized for further experiments. The patterns of phenolic change in the apoplast were compared for tobacco inoculated with P. syringae pathovars, pv. syringae, which causes a resistant HR reaction within 15 h, and pv. tabaci, which causes a susceptible reaction with delayed visible symptoms. Both pathogens elicited lower increased levels of acetosyringone compared to the saprophyte, P. fluorescens but had greatly increased levels of the chlorogenic acid derivatives. The latter metabolites appear to have come from the intracellular stores, which could

  18. Pongamia pinnata inoculated with Bradyrhizobium liaoningense PZHK1 shows potential for phytoremediation of mine tailings.

    Science.gov (United States)

    Yu, Xiumei; Li, Yangxin; Li, Yanmei; Xu, Chaohua; Cui, Yongliang; Xiang, Quanju; Gu, Yunfu; Zhao, Ke; Zhang, Xiaoping; Penttinen, Petri; Chen, Qiang

    2017-02-01

    Mine tailings contain high concentrations of metal contaminants and only little nutrients, making the tailings barren for decades after the mining has been terminated. Effective phytoremediation of mine tailings calls for deep-rooted, metal accumulating, and soil fertility increasing plants with tolerance against harsh environmental conditions. We assessed the potential of the biofuel leguminous tree Pongamia pinnata inoculated with plant growth promoting rhizobia to remediate iron-vanadium-titanium oxide (V-Ti magnetite) mine tailing soil by pot experiment and in situ remediation test. A metal tolerant rhizobia strain PZHK1 was isolated from the tailing soil and identified as Bradyrhizobium liaoningense by phylogenetic analysis. Inoculation with PZHK1 increased the growth of P. pinnata both in V-Ti magnetite mine tailings and in Ni-contaminated soil. Furthermore, inoculation increased the metal accumulation capacity and superoxide dismutase activity of P. pinnata. The concentrations of Ni accumulated by inoculated plants were higher than the hyperaccumulator threshold. Inoculated P. pinnata accumulated high concentration of Fe, far exceeding the upper limit (1000 mg kg(-1)) of Fe in plant tissue. In summary, P. pinnata-B. liaoningense PZHK1 symbiosis showed potential to be applied as an effective phytoremediation technology for mine tailings and to produce biofuel feedstock on the marginal land.

  19. Reforestation of bauxite mine spoils with Eucalyptus tereticornis Sm. seedlings inoculated with arbuscular mycorrhizal fungi

    Directory of Open Access Journals (Sweden)

    N. Krishnakumar

    2012-11-01

    Full Text Available Open cast mining for bauxite at Yercaud hills (India resulted indegradation of forest ecosystem and production of large quantities of waste rocks (called mine spoils. To ameliorate mine spoils, topsoil is used to spread over before the planting of tree species, conventional method as the topsoil has a good structure, water holding capacity and beneficial microbes like Arbuscular Mycorrhizal (AM fungi essential for plant growth.However, the use of top soil is expensive and in this study bauxite mine spoils were reforestated with AM fungi instead of it. The beneficial microbes AM fungi (Glomus aggregatum Schenck & Smith, G. fasciculatum(Thatcher Gerd. & Trappe emend. Walker & Koske, G. geosporum(Nicol. & Gerd. Walker were isolated, cultured and inoculated into the seedlings of Eucalyptus tereticornis Sm. and grown in bauxite mine spoils as potting medium under nursery conditions. Then, the biomass improved seedlings of E. tereticornis with inoculation of AM fungi were directly transplanted at bauxite mine spoils. After transplantation of the seedlings at bauxite mine spoils, the growth and survival rate were monitored for two years. The AM fungi inoculated seedlings of E. tereticornis showed 95% survival over the control seedlings and their growth was also significantlyhigher. Tissue nutrients (N, P, K were also found higher inAM fungi inoculated E. tereticornis than un inoculated control seedlings.

  20. Messenger RNA expression of chicken CLOCK gene in the response to Campylobacter jejuni inoculation.

    Science.gov (United States)

    Liu, Xiaoyi; Liu, Liying; Zhang, Maozhi; Yang, Ning; Qi, Yukai; Sun, Yu; Li, Xianyao

    2015-09-01

    Campylobacter jejuni (C. jejuni) is a leading cause of human bacterial gastroenteritis worldwide. Previous research has shown that circadian rhythm plays a critical role in host response to C. jejuni colonization. The CLOCK gene is one of the core genes regulating circadian rhythms and shows significant expression on 7 d post-C. jejuni inoculation. The objective of this study was to investigate temporal and spatial expression of chicken CLOCK gene post-C. jejuni inoculation. Cecal and splenic RNA were isolated from 2 distinct chicken breeds and used to compare the mRNA expression of CLOCK gene between inoculated and noninoculated chickens within each breed and between breeds within each of inoculated and noninoculated groups. Our results showed that the CLOCK gene was significantly down-regulated at 20 h postinoculation (hpi) in cecum and spleen in Jiningbairi chicken. CLOCK gene was significantly down-regulated at 4 and 16 hpi and up-regulated at 8 hpi in cecum and spleen in specific pathogen free white leghorn noninoculated chicken. The findings suggested that expression of CLOCK gene was significantly changed post C. jejuin inoculation. This change was affected by genetic background, tissue, and time points postinoculation. © 2015 Poultry Science Association Inc.

  1. Fatal pneumonia following inoculation of healthy bighorn sheep with Pasteurella haemolytica from healthy domestic sheep.

    Science.gov (United States)

    Foreyt, W J; Snipes, K P; Kasten, R W

    1994-04-01

    In a series of three experiments, isolates of Pasteurella haemolytica biotype A, serotype 2, ribotype reference WSU-1, from healthy domestic sheep, were inoculated intratracheally into eight bighorn sheep (Ovis canadensis canadensis) and seven domestic sheep with doses of bacteria ranging from 5.3 x 10(8) to 8.6 x 10(11) colony forming units. Seven of eight inoculated bighorn sheep died from acute pneumonia within 48 hr of inoculation, whereas all seven domestic sheep inoculated with comparable or greater doses of bacteria remained healthy. One contact control bighorn sheep also died 6 days after its penmates received P. haemolytica. Three other noncontact control bighorn sheep remained healthy during the experiments. Pasteurella haemolytica biotype A, serotype 2, ribotype reference WSU-1 in the inocula was recovered from one or more tissues from all bighorns that died; whereas, it was not detected in any bighorn sheep before inoculation. Three different ribotypes of P. haemolytica A2 were recovered from bighorn sheep; however, only the ribotype reference WSU-1 in the domestic sheep-origin inoculum was recovered from all dead bighorn sheep, and was not recovered from bighorn sheep that survived the experiments. Thus, a relatively nonpathogenic and common isolate of P. haemolytica from healthy domestic sheep was lethal in bighorn sheep under experimental conditions.

  2. Atmospheric pressure plasma jet treatment of Salmonella Enteritidis inoculated eggshells.

    Science.gov (United States)

    Moritz, Maike; Wiacek, Claudia; Koethe, Martin; Braun, Peggy G

    2017-03-20

    Contamination of eggshells with Salmonella Enteritidis remains a food safety concern. In many cases human salmonellosis within the EU can be traced back to raw or undercooked eggs and egg products. Atmospheric pressure plasma is a novel decontamination method that can reduce a wide range of pathogens. The aim of this work was to evaluate the possibility of using an effective short time cold plasma treatment to inactivate Salmonella Enteritidis on the eggshell. Therefore, artificially contaminated eggshells were treated with an atmospheric pressure plasma jet under different experimental settings with various exposure times (15-300s), distances from the plasma jet nozzle to the eggshell surface (5, 8 or 12mm), feed gas compositions (Ar, Ar with 0.2, 0.5 or 1.0% O2), gas flow rates (5 and 7slm) and different inoculations of Salmonella Enteritidis (10(1)-10(6)CFU/cm(2)). Atmospheric pressure plasma could reduce Salmonella Enteritidis on eggshells significantly. Reduction factors ranged between 0.22 and 2.27 log CFU (colony-forming units). Exposure time and, particularly at 10(4)CFU/cm(2) inoculation, feed gas had a major impact on Salmonella reduction. Precisely, longer exposure times led to higher reductions and Ar as feed gas was more effective than ArO2 mixtures.

  3. Optimization of microwave-assisted extraction for six inorganic and organic arsenic species in chicken tissues using response surface methodology.

    Science.gov (United States)

    Zhang, Wenfeng; Hu, Yuanan; Cheng, Hefa

    2015-09-01

    Response surface methodology was applied to optimize the parameters for microwave-assisted extraction of six major inorganic and organic arsenic species (As(III), As(V), dimethyl arsenic acid, monomethyl arsenic acid, p-arsanilic acid, and roxarsone) from chicken tissues, followed by detection using a high-performance liquid chromatography with inductively coupled mass spectrometry detection method, which allows the simultaneous analysis of both inorganic and organic arsenic species in the extract in a single run. Effects of extraction medium, solution pH, liquid-to-solid ratio, and the temperature and time of microwave-assisted extraction on the extraction of the targeted arsenic species were studied. The optimum microwave-assisted extraction conditions were: 100 mg of chicken tissue, extracted by 5 mL of 22% v/v methanol, 90 mmol/L (NH4 )2 HPO4 , and 0.07% v/v trifluoroacetic acid (with pH adjusted to 10.0 by ammonium hydroxide solution), ramping for 10 min to 71°C, and holding for 11 min. The method has good extraction performance for total arsenic in the spiked and nonspiked chicken tissues (104.0 ± 13.8% and 91.6 ± 7.8%, respectively), except for the ones with arsenic contents close to the quantitation limits. Limits of quantitation (S/N = 10) for As(III), As(V), dimethyl arsenic acid, monomethyl arsenic acid, p-arsanilic acid, and roxarsone in chicken tissues using this method were 0.012, 0.058, 0.039, 0.061, 0.102, and 0.240 mg/kg (dry weight), respectively.

  4. The inoculation method affects colonization and performance of bacterial inoculant strains in the phytoremediation of soil contaminated with diesel oil.

    Science.gov (United States)

    Afzal, Muhammad; Yousaf, Sohail; Reichenauer, Thomas G; Sessitsch, Angela

    2012-01-01

    Plants in combination with microorganisms can remediate soils, which are contaminated with organic pollutants such as petroleum hydrocarbons. Inoculation of plants with degrading bacteria is one approach to improve remediation processes, but is often not successful due to the competition with resident microorganisms. It is therefore of high importance to address the persistence and colonization behavior of inoculant strains. The objective of this study was to determine whether the inoculation method (seed imbibement and soil inoculation) influences bacterial colonization, plant growth promotion and hydrocarbon degradation. Italian ryegrass was grown in non-sterilized soil polluted with diesel and inoculated with different alkane-degrading strains Pantoea sp. ITSI10, Pantoea sp. BTRH79 and Pseudomonas sp. MixRI75 individually as well as in combination. Inoculation generally had a beneficial effect on plant biomass production and hydrocarbon degradation, however, strains inoculated in soil performed better than applied by seed imbibement. Performance correlated with the colonization efficiency of the inoculated strains. The highest hydrocarbon degradation was observed in the treatment, in which all three strains were inoculated in combination into soil. Our study revealed that besides the degradation potential and competitive ability of inoculant strains the inoculation method plays an important role in determining the success of microbial inoculation.

  5. Photographic-Based Optical Evaluation of Tissues and Biomaterials Used for Corneal Surface Repair: A New Easy-Applied Method.

    Directory of Open Access Journals (Sweden)

    Miguel Gonzalez-Andrades

    Full Text Available Tissues and biomaterials used for corneal surface repair require fulfilling specific optical standards prior to implantation in the patient. However, there is not a feasible evaluation method to be applied in clinical or Good Manufacturing Practice settings. In this study, we describe and assess an innovative easy-applied photographic-based method (PBM for measuring functional optical blurring and transparency in corneal surface grafts.Plastic compressed collagen scaffolds (PCCS and multilayered amniotic membranes (AM samples were optically and histologically evaluated. Transparency and image blurring measures were obtained by PBM, analyzing photographic images of a standardized band pattern taken through the samples. These measures were compared and correlated to those obtained applying the Inverse Adding-Doubling (IAD technique, which is the gold standard method.All the samples used for optical evaluation by PBM or IAD were histological suitable. PCCS samples presented transmittance values higher than 60%, values that increased with increasing wavelength as determined by IAD. The PBM indicated that PCCS had a transparency ratio (TR value of 80.3 ± 2.8%, with a blurring index (BI of 50.6 ± 4.2%. TR and BI obtained from the PBM showed a high correlation (ρ>|0.6| with the diffuse transmittance and the diffuse reflectance, both determined using the IAD (p<0.005. The AM optical properties showed that there was a largely linear relationship between the blurring and the number of amnion layers, with more layers producing greater blurring.This innovative proposed method represents an easy-applied technique for evaluating transparency and blurriness of tissues and biomaterials used for corneal surface repair.

  6. Scratching the surface: a review of skin and soft tissue infections in children.

    Science.gov (United States)

    Fenster, Daniel B; Renny, Madeline H; Ng, Carrie; Roskind, Cindy G

    2015-06-01

    We present data from recently conducted research on the diagnosis and management of skin and soft tissue infections (SSTIs) in children. Current research in the area of SSTIs (cellulitis and abscess) has focused on the use of ultrasound, risk factors associated with bacteremia, antibiotic choice, and incision and drainage (I&D) practices. When clinical examination is equivocal at distinguishing abscess from cellulitis, ultrasound can aid in the diagnosis and alter management. Bacteremia is rare in immunocompetent children with uncomplicated SSTIs; blood cultures may be reserved for complicated cases and for those who are systemically ill. Despite the increased prevalence of community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA), I&D without antibiotics remains the first-line therapy for abscess. Antibiotics for uncomplicated cellulitis should target β-hemolytic streptococci and methicillin-susceptible S. aureus (MSSA). There are significant variations in pain and sedation practices for I&D, with substantive evidence for the use of topical anesthetics. Wound packing after I&D may not confer significant benefit. Evidence to aid in the diagnosis and management of SSTIs in children has emerged in recent years; however, larger prospective pediatric studies are needed.

  7. Intensity of Ground Cover Crop Arachis pintoi, Rhizobium Inoculation and Phosphorus Application and Their Effects on Field Growth and Nutrient Status of Cocoa Plants

    Directory of Open Access Journals (Sweden)

    John Bako Baon

    2006-08-01

    Full Text Available Arachis pintoiis potentially as a cover crop for cocoa (Theobroma cacaoL. farm, however information regarding its effect on the growth of cocoa plants in the field is very limited. The objective of this experiment is to investigate the combined influence of ground cover crop A. pintoi, rhizobial bacterial inoculation and phosphorus (P fertilizer on the growth of cocoa in the field and nutrient status. This experiment laid out in split-split plot design consisted of three levels of cover crop (without, A. pintoiand Calopogonium caeruleum, two levels of rhizobium inoculation (not inoculated and inoculated and two levels of phosphorus application (no P added and P added. The results showed that in field condition the presence of A. pintoias cover crop did not affect the growth of cocoa. On the other hand, C. caeruleumas cover crop tended to restrict cocoa growth compared to A. pintoi. Application of P increased leaf number of cocoa plant. Biomass production of A. pintoiwas 40% higher than C. caeruleum. Soil organic carbon and nitrogen contents were not affected by ground cover crops, though higher value (0.235% N and 1.63% organic C was obtained from combined treatments of inoculation and P addition or neither inoculation nor P addition. In the case of no rhizobium inoculation, soil N content in cocoa farm with A. pintoicover crop was lower than that of without cover crop or with C. caeruleum. Cover crop increased plant N content when there was no inoculation, on the other hand rhizobium inoculation decreased N content of cocoa tissue. Tissue P content of cocoa plant was not influenced by A. Pintoicover crop or by rhizobium inoculation, except that the P tissue content of cocoa was 28% higher when the cover crop was C. caeruleumand inoculated. Key words : Arachis pintoi, Theobroma cacao, Calopogonium caeruleum, rhizobium, nitrogen, phosphorus.

  8. Apoplastic polyesters in Arabidopsis surface tissues--a typical suberin and a particular cutin.

    Science.gov (United States)

    Franke, Rochus; Briesen, Isabel; Wojciechowski, Tobias; Faust, Andrea; Yephremov, Alexander; Nawrath, Christiane; Schreiber, Lukas

    2005-11-01

    Cutinized and suberized cell walls form physiological important plant-environment interfaces as they act as barriers limiting water and nutrient loss and protect from radiation and invasion by pathogens. Due to the lack of protocols for the isolation and analysis of cutin and suberin in Arabidopsis, the model plant for molecular biology, mutants and transgenic plants with a defined altered cutin or suberin composition are unavailable, causing that structure and function of these apoplastic barriers are still poorly understood. Transmission electron microscopy (TEM) revealed that Arabidopsis leaf cuticle thickness ranges from only 22 nm in leaf blades to 45 nm on petioles, causing the difficulty in cuticular membrane isolation. We report the use of polysaccharide hydrolases to isolate Arabidopsis cuticular membranes, suitable for depolymerization and subsequent compositional analysis. Although cutin characteristic omega-hydroxy acids (7%) and mid-chain hydroxylated fatty acids (8%) were detected, the discovery of alpha,omega-diacids (40%) and 2-hydroxy acids (14%) as major depolymerization products reveals a so far novel monomer composition in Arabidopsis cutin, but with chemical analogy to root suberin. Histochemical and TEM analysis revealed that suberin depositions were localized to the cell walls in the endodermis of primary roots and the periderm of mature roots of Arabidopsis. Enzyme digested and solvent extracted root cell walls when subjected to suberin depolymerization conditions released omega-hydroxy acids (43%) and alpha,omega-diacids (24%) as major components together with carboxylic acids (9%), alcohols (6%) and 2-hydroxyacids (0.1%). This similarity to suberin of other species indicates that Arabidopsis roots can serve as a model for suberized tissue in general.

  9. Medical applications of VIS/NIR spectroscopy of human tissue surfaces by a novel portable instrumentation

    Science.gov (United States)

    Otto, Thomas; Stock, Volker; Schmidt, Wolf-Dieter; Liebold, Kristin; Fassler, Dieter; Wollina, Uwe; Fritzsch, Uwe; Gessner, Thomas

    2001-11-01

    In medical diagnostics, non invasive optical techniques will become common at a variety of applications because they contribute to objectivity and precision. The spectral properties of human tissue are an important field of interest. They offer opportunities of detection of skin diseases and of evaluation of chronic wounds. In the visible range, the hemoglobin absorption corresponds to blood microcirculation and the melanin absorption to the skin-type. Two types of diode-array equipment will be described: a combined VIS-NIR spectrometer system from J&M Aalen/Germany (400 nm to 1600 nm) and a stand-alone spectrometer from COLOUR CONTROL Farbmesstechnik Chemnitz/Germany (400 nm to 1000 nm). Non-contacting sensing is essential for investigating chronic wounds (no disturbances of blood microcirculation by contact pressure). The spectroscopic VIS-NIR readings of chronic wounds mainly depend on the absorption of hemoglobin and water. Multivariate analysis was applied for an objective spectral classification of eight different wound scores. Some results regarding spectral measurements of wounds and skin will be discussed. The spectrometer of COLOUR CONTROL was tested in dental surgery. To select dentures, its color has to be determined exactly to meet beauty culture demands. Color determination by dentist is not sufficient enough because of possible metameric effects of illumination. Results of spectral evaluation of denture material and human teeth will be given. Medical examination requires portable and ease equipment suitable for precise measurements. This is solved by a modular measurement system comprising microcomputer, display, light source, fiber probe, and diode-array spectrometer. It is efficient to process primary spectral data to appropriate medical interpretations.

  10. Inoculation of aluminium with titanium and boron addition

    Directory of Open Access Journals (Sweden)

    J. Szajnar

    2007-07-01

    Full Text Available Purpose: The main aim of studies was to determine mechanism of inocualtion of pure alumnium EN AW-Al99,5 structure with addition of titanium and boron, which are introduced to liquid metal in small ammount - less than obligatory standard PN-EN 573-3 (concerning about aluminium purity.Design/methodology/approach: To investigations it was used light microscopy, X-ray Phase Analysis and TEM. Surfaces of samples which were prepared for macrostructure analysis were etched with use of solution of: 50g Cu, 400ml HCl, 300ml HNO3 and 300ml H2O. Surfaces of samples which were were prepared for microstructure analysis were etched with use of solution of: 0,5ml HF, 99,5ml H2O. Thin foils for TEM investigations were electropolished with use of 20 ml HClO4 and 80ml CH3OH.Findings: The results of investigations and their analysis show, that increase of size reduction in aluminium EN AW-Al99,5 after inoculation with (Ti+B result from “washers” of type Ti3Al and CuTi2 to heterogeneous nucleation formation.Research limitations/implications: I further research, authors of this paper are going to identifiy the “washers” to heterogeneous nucleation, in aluminium structure after inoculation with zirconium and vanadium addition.Practical implications: The work presents refinement of structure method which are particularly important in continuous and semi – continuous casting where products are used for plastic forming. Large columnar crystals zone result in forces extrusion rate reduction and during the ingot rolling delamination of external layers can occur. Thus, in some cases ingot skinning is needed, which rises the production costs.Originality/value: Contributes to research on size reduction in pure aluminium structure.

  11. Examination of Some Pesticide Residues in Surface Water, Sediment and Fish Tissue of Elechi Creek, Niger Delta, Nigeria

    Directory of Open Access Journals (Sweden)

    F. Upadhi

    2012-11-01

    Full Text Available An investigation into the levels of selected pesticides in surface waters, sediment and fish (Mudskipper in the Elechi creek was carried out in two seasons and at three pre-determined sites. A higher concentration of 2, 4-diamine was observed in all stations and matrixes, while propoxur was least observed. Pesticide concentration in sediment is significantly different from that of water and fish tissue, which had the highest residual concentration. Concentration of pesticides ranged from 0.01 to 0.04 μg/L in water, 0.01 to 0.06 μg/gdw in sediment and 0.01 to 0.07 μg/gdw in fish, respectively. There was bioaccumulation of pesticides in fish samples. Observed residues are generally higher than stipulated limit of 0.01 μg/L by USEPA for pesticides of aquatic life, therefore, possess an ecological risk to the ecosystem and consequently human health.

  12. Atmospheric Cold Plasma Inactivation of Escherichia Coli, Salmonella Enterica Serovar Typhimurium and Listeria Monocytogenes Inoculated on Fresh Produce

    OpenAIRE

    2014-01-01

    Atmospheric cold plasma (ACP) represents a potential alternative to traditional methods for non-thermal decontamination of foods. In this study, the antimicrobial efficacy of a novel dielectric barrier discharge ACP device against Escherichia coli, Salmonella enterica Typhimurium and Listeria monocytogenes inoculated on cherry tomatoes and strawberries, was examined. Bacteria were spot inoculated on the produce surface, air dried and sealed inside a rigid polypropylene container. Samples were...

  13. Characterization of heparin-binding site of tissue transglutaminase: its importance in cell surface targeting, matrix deposition, and cell signaling.

    Science.gov (United States)

    Wang, Zhuo; Collighan, Russell J; Pytel, Kamila; Rathbone, Daniel L; Li, Xiaoling; Griffin, Martin

    2012-04-13

    Tissue transglutaminase (TG2) is a multifunctional Ca(2+)-activated protein cross-linking enzyme secreted into the extracellular matrix (ECM), where it is involved in wound healing and scarring, tissue fibrosis, celiac disease, and metastatic cancer. Extracellular TG2 can also facilitate cell adhesion important in wound healing through a nontransamidating mechanism via its association with fibronectin, heparan sulfates (HS), and integrins. Regulating the mechanism how TG2 is translocated into the ECM therefore provides a strategy for modulating these physiological and pathological functions of the enzyme. Here, through molecular modeling and mutagenesis, we have identified the HS-binding site of TG2 (202)KFLKNAGRDCSRRSSPVYVGR(222). We demonstrate the requirement of this binding site for translocation of TG2 into the ECM through a mechanism involving cell surface shedding of HS. By synthesizing a peptide NPKFLKNAGRDCSRRSS corresponding to the HS-binding site within TG2, we also demonstrate how this mimicking peptide can in isolation compensate for the RGD-induced loss of cell adhesion on fibronectin via binding to syndecan-4, leading to activation of PKCα, pFAK-397, and ERK1/2 and the subsequent formation of focal adhesions and actin cytoskeleton organization. A novel regulatory mechanism for TG2 translocation into the extracellular compartment that depends upon TG2 conformation and the binding of HS is proposed.

  14. Non-invasive chemically specific measurement of subsurface temperature in biological tissues using surface-enhanced spatially offset Raman spectroscopy.

    Science.gov (United States)

    Gardner, Benjamin; Stone, Nicholas; Matousek, Pavel

    2016-06-23

    Here we demonstrate for the first time the viability of characterising non-invasively the subsurface temperature of SERS nanoparticles embedded within biological tissues using spatially offset Raman spectroscopy (SORS). The proposed analytical method (T-SESORS) is applicable in general to diffusely scattering (turbid) media and features high sensitivity and high chemical selectivity. The method relies on monitoring the Stokes and anti-Stokes bands of SERS nanoparticles in depth using SORS. The approach has been conceptually demonstrated using a SORS variant, transmission Raman spectroscopy (TRS), by measuring subsurface temperatures within a slab of porcine tissue (5 mm thick). Root-mean-square errors (RMSEs) of 0.20 °C were achieved when measuring temperatures over ranges between 25 and 44 °C. This unique capability complements the array of existing, predominantly surface-based, temperature monitoring techniques. It expands on a previously demonstrated SORS temperature monitoring capability by adding extra sensitivity stemming from SERS to low concentration analytes. The technique paves the way for a wide range of applications including subsurface, chemical-specific, non-invasive temperature analysis within turbid translucent media including: the human body, subsurface monitoring of chemical (e.g. catalytic) processes in manufacture quality and process control and research. Additionally, the method opens prospects for control of thermal treatment of cancer in vivo with direct non-invasive feedback on the temperature of mediating plasmonic nanoparticles.

  15. Establishment of a leptospirosis model in guinea pigs using an epicutaneous inoculations route

    Directory of Open Access Journals (Sweden)

    Zhang Yan

    2012-01-01

    Full Text Available Abstract Background Leptospires are presumed to enter their host via small abrasions or breaches of the skin. The intraperitoneal route, although commonly used in guinea pig and hamster models of leptospirosis, does not reflect conditions encountered during natural infection. The aim of this study is to develop a novel leptospirosis guinea pig model through epicutaneous route and to elucidate the pathogenesis of leptospirosis in experimental guinea pigs by comparing the data from other studies using different infection routes. Methods The guinea pigs were inoculated with 5 × 108 Leptospira interrogans strain Lai onto either shaved-only or abraded skin. The guinea pigs were sacrificed at 2, 8, 24, 48, 72, 96 and 144 h post-infection (p.i. followed by harvest of the lungs, liver, kidneys, spleen, and the skin around the inoculated sites for further examinations. Hematoxylin and eosin (HE staining and electron microscopy were used to detect the pathologic changes. Real time PCR and immunohistochemistry staining were performed to detect dynamic distribution of leptospires in blood and tissues, respectively. Results In the guinea pigs with abraded skin inoculations, leptospires were detected in blood as early as 2 h post infection (p.i. and then disseminated to the liver, lungs and kidneys of almost all animals within 96 h p.i.. Leptospires were also detected engulfed in the swelling vascular endothelial cells and were frequently aggregated around the capillaries in the dermis and subcutaneous tissue under the inoculated site. For the guinea pigs with abraded skin inoculations, hemorrhage at the dermis around the inoculated site was found before the appearance of internal organs hemorrhage, severe lesions such as hemorrhages in the lungs, nephritis, jaundice, haematuria were also observed, and two of seven guinea pigs died at 144 h p.i. while no lesions and leptospires were detected in the shaved-only guinea pigs using the same dose of strain Lai

  16. Inoculant of Arbuscular Mycorrhizal Fungi (Rhizophagus clarus) Increase Yield of Soybean and Cotton under Field Conditions

    Science.gov (United States)

    Cely, Martha V. T.; de Oliveira, Admilton G.; de Freitas, Vanessa F.; de Luca, Marcelo B.; Barazetti, André R.; dos Santos, Igor M. O.; Gionco, Barbara; Garcia, Guilherme V.; Prete, Cássio E. C.; Andrade, Galdino

    2016-01-01

    Nutrient availability is an important factor in crop production, and regular addition of chemical fertilizers is the most common practice to improve yield in agrosystems for intensive crop production. The use of some groups of microorganisms that have specific activity providing nutrients to plants is a good alternative, and arbuscular mycorrhizal fungi (AMF) enhance plant nutrition by providing especially phosphorus, improving plant growth and increasing crop production. Unfortunately, the use of AMF as an inoculant on a large scale is not yet widely used, because of several limitations in obtaining a large amount of inoculum due to several factors, such as low growth, the few species of AMF domesticated under in vitro conditions, and high competition with native AMF. The objective of this work was to test the infectivity of a Rhizophagus clarus inoculum and its effectiveness as an alternative for nutrient supply in soybean (Glycine max L.) and cotton (Gossypium hirsutum L.) when compared with conventional chemical fertilization under field conditions. The experiments were carried out in a completely randomized block design with five treatments: Fertilizer, AMF, AMF with Fertilizer, AMF with 1/2 Fertilizer, and the Control with non-inoculated and non-fertilized plants. The parameters evaluated were AMF root colonization and effect of inoculation on plant growth, nutrient absorption and yield. The results showed that AMF inoculation increased around 20 % of root colonization in both soybean and cotton; nutrients analyses in vegetal tissues showed increase of P and nitrogen content in inoculated plants, these results reflect in a higher yield. Our results showed that, AMF inoculation increase the effectiveness of fertilizer application in soybean and reduce the fertilizer dosage in cotton. PMID:27303367

  17. Fading of inoculation effects in ductile iron

    Directory of Open Access Journals (Sweden)

    E. Fraś

    2008-03-01

    Full Text Available In work i t has bccn shown rcsults or invcsligations of influcncc of rime Iapsed form inoculation proccss on graphitc nucleation potentialrcprcscntcd by: numbcr of graphitc nodulcs N and N,, maximum undercooling AT,, during solidification of gmphile eutcct ic. abmlutcchilling tcndcncy CT and critical casting diametct dh. undcr which cementite euteclic occur (so-callcd chills. Morcovcr it has hccncstima~cd raic of changc of N and N, AT,,,. CT and dk,. Also, it has bccn provcd that altcr onc minutc sincc rhc momcnt of inocuIationproccss nhout 35% of prnphttc nucIeation potenrial is tost. by 40% chitking tendency, by 70% incrcascs maximum undcrcmling forgraphitc ci~tccrica nd by nearly 40% caging diameter has to bc incrcascd in ordcr to avoid chills.

  18. Fabrication and characterization of a novel microparticle with gyrus-patterned surface and growth factor delivery for cartilage tissue engineering

    Energy Technology Data Exchange (ETDEWEB)

    Huang Sha [Department of Oral Histology and Pathology, School of Stomatology, Fourth Military Medical University, Xi' an 710032 (China); Research and Development Center for Tissue Engineering, Fourth Military Medical University, Xi' an 710032 (China); Wang Yijuan [Key Laboratory for Macromolecular Science of Shaanxi Province, Shaanxi Normal University, Xi' an 710062 (China); Liang Tang [Department of Oral Histology and Pathology, School of Stomatology, Fourth Military Medical University, Xi' an 710032 (China); Research and Development Center for Tissue Engineering, Fourth Military Medical University, Xi' an 710032 (China); Jin Fang [Department of Orthodontics, School of Stomatology, Fourth Military Medical University, Xi' an 710032 (China); Liu Shouxin [Key Laboratory for Macromolecular Science of Shaanxi Province, Shaanxi Normal University, Xi' an 710062 (China); Jin Yan, E-mail: yanjin@fmmu.edu.cn [Department of Oral Histology and Pathology, School of Stomatology, Fourth Military Medical University, Xi' an 710032 (China); Research and Development Center for Tissue Engineering, Fourth Military Medical University, Xi' an 710032 (China)

    2009-05-05

    Microparticles can serve as substrates for cell amplification and deliver the expanded cells to the site of the defect. It was hypothesized that a novel microparticle combined of sustained and localized delivery of proliferative growth factors and gyrus-patterned surface would influence the cell behaviours of adherence and expansion on the microparticle in the present study. To test the hypothesis, gelatin particles with diameter ranging from 280 to 350 {mu}m were fabricated and were modified by cryogenic freeze-drying treatment and basic fibroblast growth factor (bFGF) incorporation. The results of in vitro chondrocyte culture illustrated that cells could proliferate more obviously on the microparticles with bFGF addition, but no correlation between attachment rate and bFGF was observed. On the other hand, microparticles with gyrus-patterned surface demonstrated the highest cell attachment rate and higher rate of cell growth, in particular on bFGF combined ones. It seems to be a promising candidate as a chondrocyte microparticle and could be the potential application in cartilage tissue engineering.

  19. Surface marker cluster translation, rotation, scaling and deformation: Their contribution to soft tissue artefact and impact on knee joint kinematics.

    Science.gov (United States)

    Benoit, D L; Damsgaard, M; Andersen, M S

    2015-07-16

    When recording human movement with stereophotogrammetry, skin deformation and displacement (soft tissue artefact; STA) inhibits surface markers' ability to validly represent the movement of the underlying bone. To resolve this issue, the components of marker motions which contribute to STA must be understood. The purpose of this study is to describe and quantify which components of this marker motion (cluster translation, rotation, scaling and deformation) contribute to STA during the stance phase of walking, a cutting manoeuvre, and one-legged hops. In vivo bone pin-based tibio-femoral kinematics of six healthy subjects were used to study skin marker-based STA. To quantify how total cluster translation, rotation, scaling and deformation contribute to STA, a resizable and deformable cluster model was constructed. STA was found to be greater in the thigh than the shank during all three movements. We found that the non-rigid (i.e. scaling and deformation) movements contribute very little to the overall amount of error, rendering surface marker optimisation methods aimed at minimising this component superfluous. The results of the current study indicate that procedures designed to account for cluster translation and rotation during human movement are required to correctly represent the motion of body segments, however reducing marker cluster scaling and deformation will have little effect on STA.

  20. Understanding the influence of surface properties of nanoparticles and penetration enhancers for improving bioavailability in eye tissues in vivo.

    Science.gov (United States)

    Mahaling, Binapani; Katti, Dhirendra S

    2016-03-30

    Nanoparticulate drug delivery systems, mucoadhesive polymers and penetration enhancers have been used individually to overcome ocular barriers and increase bioavailability to eye tissues. However the combined influence of mucoadhesive polymer coating and penetration enhancers on NP permeability has not been investigated. Hence, in this study, three types of core-shell nanoparticles with same hydrophobic core but different hydrophilic mucoadhesive shells were developed. Initially the influence of a single penetration enhancer (PE) [benzalkonium chloride (BAC)] on ocular permeability of all the three core-shell nanoparticles was studied. Then ocular permeability of a single nanoparticle system [polycaprolactone-pluronicF68 (PCL-PF68)] in presence of different types of PEs namely BAC, capric acid (CA), EDTA, sodium glycocolate (SG) and sodium taurocholate (ST) was studied. The results indicated that BAC differentially enhanced ocular permeability of nanoparticles depending on their surface properties. All the PEs except EDTA enhanced ocular permeability of PCL-PF68 nanoparticles to anterior part of the eye. BAC and CA showed increased bioavailability of PCL-PF68 nanoparticles in conjunctiva, SG in cornea, iris and ciliary body, and ST in cornea. Overall, the combination of PEs and surface properties of nanoparticles can differentially influence ocular permeability and bioavailability and can be advantageously used to develop improved ocular drug delivery systems.

  1. Concave Pit-Containing Scaffold Surfaces Improve Stem Cell-Derived Osteoblast Performance and Lead to Significant Bone Tissue Formation

    Science.gov (United States)

    Cusella-De Angelis, Maria Gabriella; Laino, Gregorio; Piattelli, Adriano; Pacifici, Maurizio; De Rosa, Alfredo; Papaccio, Gianpaolo

    2007-01-01

    Background Scaffold surface features are thought to be important regulators of stem cell performance and endurance in tissue engineering applications, but details about these fundamental aspects of stem cell biology remain largely unclear. Methodology and Findings In the present study, smooth clinical-grade lactide-coglyolic acid 85:15 (PLGA) scaffolds were carved as membranes and treated with NMP (N-metil-pyrrolidone) to create controlled subtractive pits or microcavities. Scanning electron and confocal microscopy revealed that the NMP-treated membranes contained: (i) large microcavities of 80–120 µm in diameter and 40–100 µm in depth, which we termed primary; and (ii) smaller microcavities of 10–20 µm in diameter and 3–10 µm in depth located within the primary cavities, which we termed secondary. We asked whether a microcavity-rich scaffold had distinct bone-forming capabilities compared to a smooth one. To do so, mesenchymal stem cells derived from human dental pulp were seeded onto the two types of scaffold and monitored over time for cytoarchitectural characteristics, differentiation status and production of important factors, including bone morphogenetic protein-2 (BMP-2) and vascular endothelial growth factor (VEGF). We found that the microcavity-rich scaffold enhanced cell adhesion: the cells created intimate contact with secondary microcavities and were polarized. These cytological responses were not seen with the smooth-surface scaffold. Moreover, cells on the microcavity-rich scaffold released larger amounts of BMP-2 and VEGF into the culture medium and expressed higher alkaline phosphatase activity. When this type of scaffold was transplanted into rats, superior bone formation was elicited compared to cells seeded on the smooth scaffold. Conclusion In conclusion, surface microcavities appear to support a more vigorous osteogenic response of stem cells and should be used in the design of therapeutic substrates to improve bone repair and

  2. Concordance of human papillomavirus types detected on the surface and in the tissue of genital lesions in men.

    Science.gov (United States)

    Anic, Gabriella M; Messina, Jane L; Stoler, Mark H; Rollison, Dana E; Stockwell, Heather; Villa, Luisa L; Lazcano-Ponce, Eduardo; Gage, Christine; Silva, Roberto Jose C; Baggio, Maria L; Salmerón, Jorge; Giuliano, Anna R

    2013-09-01

    Swabbing the surface of a genital lesion to obtain a sample for HPV DNA testing is less invasive than a biopsy, but may not represent HPV types present in the lesion tissue. The objective of this study was to examine the concordance of HPV types detected in swab and biopsy samples from 165 genital lesions from men ages 18-70. Lesions included 90 condyloma, 10 penile intraepithelial neoplasia (PeIN), 23 non-condyloma with a known histology, and 42 lesions with an undetermined histology. All lesions were sampled by swabbing the surface of the lesion with a pre-wetted Dacron swab and taking a shave biopsy. HPV genotyping was performed using Linear Array for swab samples and INNO-LiPA for biopsy samples. The kappa and McNemar statistics were used to compare the concordance of detecting HPV types in swab and biopsy samples. Both sampling methods had high agreement for detection of HPV DNA in condyloma (87.8% agreement) and PeIN (100% agreement). There was also high concordance for detection of HPV16 (kappa = 1.00) and HPV18 (kappa = 1.00) in PeIN, however, agreement was low to moderate for detecting HPV6 (kappa = 0.31) and HPV11 (kappa = 0.56) in condyloma. Low to moderate agreement was also observed between sampling methods for detecting individual HPV types in the non-condyloma and lesions with an indefinite histology. The results suggest that obtaining a biopsy in addition to swabbing the surface of a lesion may provide additional information about specific HVP types associated with male genital lesions. Copyright © 2013 Wiley Periodicals, Inc.

  3. Microwave-assisted fibrous decoration of mPE surface utilizing Aloe vera extract for tissue engineering applications.

    Science.gov (United States)

    Balaji, Arunpandian; Jaganathan, Saravana Kumar; Supriyanto, Eko; Muhamad, Ida Idayu; Khudzari, Ahmad Zahran Md

    2015-01-01

    Developing multifaceted, biocompatible, artificial implants for tissue engineering is a growing field of research. In recent times, several works have been reported about the utilization of biomolecules in combination with synthetic materials to achieve this process. Accordingly, in this study, the ability of an extract obtained from Aloe vera, a commonly used medicinal plant in influencing the biocompatibility of artificial material, is scrutinized using metallocene polyethylene (mPE). The process of coating dense fibrous Aloe vera extract on the surface of mPE was carried out using microwaves. Then, several physicochemical and blood compatibility characterization experiments were performed to disclose the effects of corresponding surface modification. The Fourier transform infrared spectrum showed characteristic vibrations of several active constituents available in Aloe vera and exhibited peak shifts at far infrared regions due to aloe-based mineral deposition. Meanwhile, the contact angle analysis demonstrated a drastic increase in wettability of coated samples, which confirmed the presence of active components on glazed mPE surface. Moreover, the bio-mimic structure of Aloe vera fibers and the influence of microwaves in enhancing the coating characteristics were also meticulously displayed through scanning electron microscopy micrographs and Hirox 3D images. The existence of nanoscale roughness was interpreted through high-resolution profiles obtained from atomic force microscopy. And the extent of variations in irregularities was delineated by measuring average roughness. Aloe vera-induced enrichment in the hemocompatible properties of mPE was established by carrying out in vitro tests such as activated partial thromboplastin time, prothrombin time, platelet adhesion, and hemolysis assay. In conclusion, the Aloe vera-glazed mPE substrate was inferred to attain desirable properties required for multifaceted biomedical implants.

  4. Effects of biotic and abiotic stress on induced accumulation of terpenes and phenolics in red pines inoculated with bark beetle-vectored fungus.

    Science.gov (United States)

    Klepzig, K D; Kruger, E L; Smalley, E B; Raffa, K F

    1995-05-01

    This study characterized the chemical response of healthy red pine to artificial inoculation with the bark beetle-vectored fungusLeptographium terebrantis. In addition, we sought to determine whether stress altered this induced response and to understand the implications of these interactions to the study of decline diseases. Twenty-five-year-old trees responded to mechanical wounding or inoculation withL. terebrantis by producing resinous reaction lesions in the phloem. Aseptically wounded and wound-inoculated phloem contained higher concentrations of phenolics than did constitutive tissue. Trees inoculated withL. terebrantis also contained higher concentrations of six monoterpenes,α-pinene,β-pinene, 3-carene, limonene, camphene, and myrcene, and higher total monoterpenes than did trees that were mechanically wounded or left unwounded. Concentrations of these monoterpenes increased with time after inoculation. Total phenolic concentrations in unwounded stem tissue did not differ between healthy and root-diseased trees. Likewise, constitutive monoterpene concentrations in stem phloem were similar between healthy and root-diseased trees. However, when stem phloem tissue was challenged with fungal inoculations, reaction tissue from root-diseased trees contained lower concentrations ofα-pinene, the predominant monoterpene in red pine, than did reaction tissue from healthy trees. Seedlings stressed by exposure to low light levels exhibited less extensive induced chemical changes when challenge inoculated withL. terebrantis than did seedlings growing under higher light. Stem phloem tissue in these seedlings contained lower concentrations ofα-pinene than did nonstressed seedlings also challenge inoculated withL. terebrantis. It is hypothesized that monoterpenes and phenolics play a role in the defensive response of red pine against insect-fungal attack, that stress may predispose red pine to attack by insect-fungal complexes, and that such interactions are involved

  5. Hepatocellular carcinoma. A study of 50 autopsy cases with detection of hepatitis B surface antigen in fixed tissues.

    Science.gov (United States)

    Perez-Barrios, A; Colina-Ruizdelgado, F; Gallego, I; Martinez-Tello, F J

    1983-03-01

    Fifty patients who died of hepatocellular carcinoma (HCC) were autopsied at the Ciudad Sanitaria "1 degree de Octubre" and the Hospital de la Cruz Roja (Madrid) from 1974 to 1980. Formalin fixed paraffin-embedded autopsy tissue of liver and tumor from the 50 HCC and liver tissue from 50 liver cirrhosis (LC) and from 50 autopsy of non cirrhotic control cases were examined for the presence of cytoplasmic hepatitis B surface antigen (HBsAg). The study was carried out using orcein staining, immunoperoxidase technique (IP) and indirect immunofluorescence (IF). In livers with HCC the HBsAg was detected in the cytoplasm of the hepatocytes in 10 cases (20%) with the orcein staining and in 11 (22%) with the IP and IF techniques. In one case (2%) HBsAg was found in the cytoplasm of tumor cells with the three methods--In four cases (8%) of LC and 2 (4%) control cases cytoplasmic positive cells were found. In 41 patients with HCC HBsAg was studied in the serum by radio-immunoassay (RIA) (13 cases) and immunodiffussion (28 cases). 5 patients (12,1%) were positive and 36 (72%) were negative. In the 5 serum positive HBsAg HCC the staining methods for cytoplasmic HBsAg were positive (100%). In 36 serum negative HBsAg HCC the staining method were positive in 2 cases. The results let us to conclude that HBV is a probable important etiologic factor of HCC in our milieu. 54% of the patients with HCC had a previous history of alcohol abuse; however, histologic features compatible with an alcoholic etiology were found in only 5 cases. Nevertheless we consider that the described histopathologic findings do not exclude excess alcohol consumption as a possible etiologic factor for HCC in our series.

  6. Re-epithelializaiton by epithelial inoculation with recipient phenotype in heterotopically transplanted rat allografts

    Institute of Scientific and Technical Information of China (English)

    Zheng Hui; Hu Xuefei; Li Chao; Xie Huikang; Gao Wen; Chen Chang

    2014-01-01

    Background Re-epithelialization has remained a major obstacle in both tracheal and lung transplantations.This study examines the realization of re-epithelialization by epithelial inoculation in a rat heterotopic tracheal transplantation model.Methods The original epithelia of tracheas from donor Wistar rats were removed and the tracheas were then inoculated with 106/ml in vitro cultured epithelial cells of the Spraque-Dawley (SD) rat phenotype.These allo-tracheas were then heterotopically transplanted into SD rats.After 28 days,the allo-trachea tissues were recovered and assessed for epithelial morphology and cellular differentiation using immunohistochemical analysis.An additional experimental group was used to compare the outcomes of re-epithelialization in immunosuppressed animals.Results Histological examination showed that allografts with epithelial inoculation maintained patent tracheal lumens,which were obliterated in controls.Recipient immunosuppression facilitated the formation of an integrated ciliated epithelial layer,further demonstrated by the presence of a dense cilia population,a well-developed plasma membrane,and readily recognizable intercellular junctions.Epithelial cellular differentiation markers such as cytokeratin 14 and 18,and cystic fibrosis transmembrane conductance regulator (CFTR) were all positive in allografts under immunosuppression.Conclusion Concurrent recipient-derived epithelial inoculation with immunosuppression can result in complete reepithelialization with the recipient phenotype and suppress the luminal obliteration process in heterotopic transplantations.

  7. Investigation of Figopitant and Its Metabolites in Rat Tissue by Combining Whole-Body Autoradiography with Liquid Extraction Surface Analysis Mass Spectrometry

    DEFF Research Database (Denmark)

    Schadt, S.; Kallbach, S.; Almeida, R.;

    2012-01-01

    This article describes the combination of whole-body autoradiography with liquid extraction surface analysis (LESA) and mass spectrometry (MS) to study the distribution of the tachykinin neurokinin-1 antagonist figopitant and its metabolites in tissue sections of rats after intravenous...... tissue extraction, sample cleanup, and high-performance liquid chromatography analysis. The parent drug and the N-dealkylated metabolite M474(1) (BIIF 1148) in varying ratios were the predominant compounds in all tissues investigated. In addition, several metabolites formed by oxygenation, dealkylation...

  8. Fungi Isolated from Flue-Cured Tobacco Inoculated in the Field with Storage Fungi

    Science.gov (United States)

    Welty, Ronald E.

    1971-01-01

    Flue-cured tobacco inoculated in the field with A. amstelodami, A. flavus, A. ochraceus, A. repens, A. ruber, and a species of Penicillium was rarely invaded by these fungi. Regardless of inoculum, the predominant fungi reisolated from green tissue were species of Alternaria and Cladosporium. After curing, A. repens, A. niger, and species of Alternaria and a species of Penicillium were the most commonly isolated fungi. The fungus used as inoculum was not the predominant fungus reisolated from green or cured tissue. Conditions during handling and storage prior to marketing probably determine when storage fungi become associated with the leaf and which species becomes predominant. PMID:5102779

  9. Alpha-1 proteinase inhibitor M358R reduces thrombin generation when displayed on the surface of cells expressing tissue factor.

    Science.gov (United States)

    Gierczak, Richard F; Pepler, Laura; Bhagirath, Vinai; Liaw, Patricia C; Sheffield, William P

    2014-11-01

    The M358R variant of alpha-1-proteinase inhibitor (API) is a potent soluble inhibitor of thrombin. Previously we engineered AR-API M358R, a membrane-bound form of this protein and showed that it inhibited exogenous thrombin when expressed on transfected cells lacking tissue factor (TF). To determine the suitability of AR-API M358R for gene transfer to vascular cells to limit thrombogenicity, we tested the ability of AR-API M358R to inhibit endogenous thrombin generated in plasma via co-expression co-expressing it on the surface of cells expressing TF. Transfected AR-API M358R formed inhibitory complexes with thrombin following exposure of recalcified, defibrinated plasma to TF on T24/83 cells, but discontinuously monitored thrombin generation was unaffected. Similarly, AR-API M358R expression did not reduce continuously monitored thrombin generation by T24/83 cell suspensions exposed to recalcified normal plasma in a Thrombogram-Thrombinoscope-type thrombin generation assay (TGA); in contrast, 1 μM hirudin variant 3 or soluble API M358R abolished thrombin generation. Gene transfer of TF to HEK 293 conferred the ability to support TF-dependent thrombin generation on HEK 293 cells. Co-transfection of HEK 293 cells with a 9:1 excess of DNA encoding AR-API M358R to that encoding TF reduced peak thrombin generation approximately 3-fold compared to controls. These in vitro results suggest that surface display of API M358R inhibits thrombin generation when the tethered serpin is expressed in excess of TF, and suggest its potential to limit thrombosis in appropriate vascular beds in animal models.

  10. AMF Inoculation Enhances Growth and Improves the Nutrient Uptake Rates of Transplanted, Salt-Stressed Tomato Seedlings

    Directory of Open Access Journals (Sweden)

    Astrit Balliu

    2015-12-01

    Full Text Available The study aimed to investigate the effects of commercially available AMF inoculate (Glomus sp. mixture on the growth and the nutrient acquisition in tomato (Solanumlycopersicum L. plants directly after transplanting and under different levels of salinity. Inoculated (AMF+ and non-inoculated (AMF− tomato plants were subjected to three levels of NaCl salinity (0, 50, and 100 mM·NaCl. Seven days after transplanting, plants were analyzed for dry matter and RGR of whole plants and root systems. Leaf tissue was analyzed for mineral concentration before and after transplanting; leaf nutrient content and relative uptake rates (RUR were calculated. AMF inoculation did not affect plant dry matter or RGR under fresh water-irrigation. The growth rate of AMF−plants did significantly decline under both moderate (77% and severe (61% salt stress compared to the fresh water-irrigated controls, while the decline was much less (88% and 75%,respectivelyand statistically non-significant in salt-stressed AMF+ plants. Interestingly, root system dry matter of AMF+ plants (0.098 g plant–1 remained significantly greater under severe soil salinity compared to non-inoculated seedlings (0.082 g plant–1. The relative uptake rates of N, P, Mg, Ca, Mn, and Fe were enhanced in inoculated tomato seedlings and remained higher under (moderate salt stress compared to AMF− plants This study suggests that inoculation with commercial AMF during nursery establishment contributes to alleviation of salt stress by maintaining a favorable nutrient profile. Therefore, nursery inoculation seems to be a viable solution to attenuate the effects of increasing soil salinity levels, especially in greenhouses with low natural abundance of AMF spores.

  11. Sarcocyst Development in Raccoons (Procyon lotor) Inoculated with Different Strains of Sarcocystis neurona Culture-Derived Merozoites.

    Science.gov (United States)

    Dryburgh, E L; Marsh, A E; Dubey, J P; Howe, D K; Reed, S M; Bolten, K E; Pei, W; Saville, W J A

    2015-08-01

    Sarcocystis neurona is considered the major etiologic agent of equine protozoal myeloencephalitis (EPM), a neurological disease in horses. Raccoon ( Procyon lotor ) is considered the most important intermediate host in the life cycle of S. neurona in the United States; S. neurona sarcocysts do mature in raccoon muscles, and raccoons also develop clinical signs simulating EPM. The focus of this study was to determine if sarcocysts would develop in raccoons experimentally inoculated with different host-derived strains of in vitro-cultivated S. neurona merozoites. Four raccoons were inoculated with strains derived from a raccoon, a sea otter, a cat, and a horse. Raccoon tissues were fed to laboratory-raised opossums ( Didelphis virginiana ), the definitive host of S. neurona . Intestinal scraping revealed sporocysts in opossums who received muscle tissue from raccoons inoculated with the raccoon-derived or the sea otter-derived isolates. These results demonstrate that sarcocysts can mature in raccoons inoculated with in vitro-derived S. neurona merozoites. In contrast, the horse and cat-derived isolates did not produce microscopically or biologically detected sarcocysts. Immunoblot analysis revealed both antigenic and antibody differences when testing the inoculated raccoons. Immunohistochemical staining indicated differences in staining between the merozoite and sarcocyst stages. The successful infections achieved in this study indicates that the life cycle can be manipulated in the laboratory without affecting subsequent stage development, thereby allowing further purification of strains and artificial maintenance of the life cycle.

  12. A study of parabens and bisphenol A in surface water and fish brain tissue from the Greater Pittsburgh Area.

    Science.gov (United States)

    Renz, Lara; Volz, Conrad; Michanowicz, Drew; Ferrar, Kyle; Christian, Charles; Lenzner, Diana; El-Hefnawy, Talal

    2013-05-01

    Pollution from xenoestrogens has been discovered in the aquatic environment of the Greater Pittsburgh Area and is suspected to be caused by the failing sewer system. Personal care products and plasticizers have the potential to enter the water supply though treated and untreated sewage. Many of these compounds are suspected xenoestrogens. Paraben detection in surface waters was as follows: methyl paraben ranged between 2.2 to 17.3 ppt; ethyl paraben was not detectable; propyl paraben was detected at 9.2 and 12.0 ppt; butyl paraben was detected at 0.2 ppt. BPA was detected between 0.6 and 15.4 ppt. Estrogenic potential of extracts from fish brain tissue was tested via Bromodeoxyuridine MCF-7 analysis and paired with HPLC-MS to investigate the presence of xenoestrogens. All samples were non-detectable for parabens. BPA was detected in 44 of the 58 samples, with a range from non-detectable to 120 pg/g. BCFs were calculated. Results were statistically significant for location of capture (p < 0.05) and correlation existed between estrogenicity and BPA.

  13. Preliminary observation of effects of extrace from inflammatory cutaneous tissues of the rabbit inoculated with vaccina virus in the treatment of hyperextension injuries of cervical spinal cord%牛痘疫苗接种家兔炎症皮肤提取物早期治疗颈髓过伸伤的疗效观察

    Institute of Scientific and Technical Information of China (English)

    接健; 陈福扬; 冯玉旭; 徐国浩; 王梅生

    2014-01-01

    Objective To explore the preliminary clinical effects of extrace from inlfammatory cutaneous tissues of the rabbit inoculated with vaccina virus, also called Neurotropin ( NTP ) in the treatment of hyperextension injuries of cervical spinal cord. Methods From January 2010 to January 2014, 20 patients with hyperextension injuries of cervical spinal cord were adopted, who were randomly divided into the NTP group and the control group. Traction was performed on the patients in the NTP group for 3 weeks and then plaster ifxation for 3 months, besides the NTP treatment. Occipital-jaw-traction was performed on the patients in the control group for 3 weeks and then plaster fixation for 3 months. The Visual Analogue Scale ( VAS ) was used to assess pain, limb numbness and electrified sensation in both groups, and the scores before the treatment and at the end of traction and treatment were recorded and compared. Results The VAS scores of pain, limb numbness and electriifed sensation in the NTP group were ( 6.2±1.81 ) points, ( 5.3±1.15 ) points and ( 5.4±0.84 ) points before the treatment, which were obviously reduced to ( 1.2±0.78 ) points, ( 1.6±0.84 ) points and ( 0.6±0.51 ) points at the end of traction. The differences between them were statistically signiifcant ( P0.05 ). The VAS scores of pain, limb numbness and electriifed sensation in the NTP group were ( 1.3±0.98 ) points, ( 1.8±0.86 ) points and ( 0.5±0.49 ) points at the end of treatment, which were respectively lower than ( 2.6±0.49 ) points, ( 2.9±1.26 ) points and ( 1.8±0.76 ) points in the control group. The differences between them were statistically signiifcant ( P<0.05 ). Cervical spine alignment was restored in all the patients when the plaster was removed, and the cervical spine was stable. No cervical unstability, pneumonia or bedsore occurred. According to the Frankel grading system, 2 of the 3 patients rated as grade A underwent NTP treatment, and the last one did not. The improvement in

  14. Real-Time Monitoring of Escherichia coli O157:H7 Adherence to Beef Carcass Surface Tissues with a Bioluminescent Reporter

    OpenAIRE

    Siragusa, Gregory R.; Nawotka, Kevin; Spilman, Stanley D.; Contag, Pamela R.; Christopher H. Contag

    1999-01-01

    A method for studying bacteria that are attached to carcass surfaces would eliminate the need for exogenous sampling and would facilitate understanding the interaction of potential human food-borne pathogens with food animal tissue surfaces. We describe such a method in which we used a bioluminescent reporter strain of Escherichia coli O157:H7 that was constructed by transformation with plasmid pCGLS1, an expression vector that contains a complete bacterial luciferase (lux) operon. Beef carca...

  15. Preliminary attempts to biolistic inoculation of grapevine fanleaf virus.

    Science.gov (United States)

    Valat, L; Mode, F; Mauro, M C; Burrus, M

    2003-03-01

    Biolistics has been studied to inoculate grapevine fanleaf virus (GFLV), a Nepovirus, to its natural woody host, Vitis sp., and its herbaceous host, Chenopodium quinoa. At first, bombardment conditions for in vitro and greenhouse grown plants were set using the uidA reporter gene. The infectious feature of the cartridges was then evaluated by studying infection of C. quinoa plants. Systemic infection was obtained with either GFLV particles or RNA extracts in experimental conditions which gave also the highest transient uidA gene expression. Concerning grapevine, our results indicate that extrapolation to this plant is difficult. In only 1 out of 8 independent bombardment experiments done with GFLV and 41B, we were able to detect the virus in freshly bombarded leaves. Similarly, later after bombardment, Pol mRNAs were detected once, at days 7 and 14 only. Incubating the plants in darkness, as suggested in the literature, or using Rupestris Saint Georges, an indicator for GFLV presence, did not yield any improvement. Finely, our observations suggest that detection of GFLV in bombarded grapevine tissues by immunological or molecular techniques remains a limiting factor, probably due to an excess of inhibitory compounds released during the biolistic process.

  16. Effects of genotype x bradyrhizobium inoculation or x fertilizer n ...

    African Journals Online (AJOL)

    Genotype x inoculation interaction was significant only for protein and oil content in one year. ... seed composition, or other agronoi:nic traits including plant .... used to compare expected genetic of inoculation on maturity, p;ant height, a;:d yick!

  17. Silage Inoculant Effects on In Vitro Rumen Fermentation

    Science.gov (United States)

    Four inoculants, B (Lactobacillus plantarum and Enterococcus faecium), C (Lactobacillus plantarum), D (Lactobacillus pentosus), E (Lactococcus lactis), were compared with an uninoculated treatment (A) on alfalfa (38% DM, AS), corn (36% DM, CS), and brown midrib corn (33% DM, BMR) silages. All inocul...

  18. Lactating cow response to lucerne silage inoculated with Lactobacillus plantarum

    Science.gov (United States)

    It is unclear why bacterial silage inoculants improve milk production in lactating dairy cattle. However, recent in vitro results suggest that inoculated silage effects on milk production may be tied to greater production of rumen microorganisms. Our objective was to determine if alfalfa silage trea...

  19. Comparison of a modified shell vial culture procedure with conventional mouse inoculation for rabies virus isolation

    Directory of Open Access Journals (Sweden)

    María de los Angeles Ribas Antúnez

    2013-04-01

    Full Text Available Rabies is a neurotropic disease that is often lethal. The early diagnosis of rabies infection is important and requires methods that allow for the isolation of the virus from animals and humans. The present study compared a modified shell vial (MSV procedure using 24-well tissue culture plates with the mouse inoculation test (MIT, which is considered the gold standard for rabies virus isolation. Thirty brain samples (25 positive and 5 negative by the fluorescent antibody test obtained from different animal species at the National Institute of Hygiene Rafael Rangel in Caracas, Venezuela, were studied by the MIT and MSV assays. Nine samples (36% were positive at 24 h, 10 (40% were positive at 48 h and six (24% were positive at 72 h by the MSV assay. With the MIT assay, 76% were positive at six days post inoculation and 12% were positive at 12 and 18 days post inoculation. One sample that was negative according to the MSV assay was positive with MIT on the 12th day. The MSV procedure exhibited a sensitivity of 96.2%, a specificity of 100%, a positive predictive value of 100% and a negative predictive value 80%. This procedure allowed for rapid rabies virus detection. MIT can be employed as an alternative method in laboratories without tissue culture facilities.

  20. Microbial Inoculation Improves Growth of Oil Palm Plants (Elaeis guineensis Jacq.).

    Science.gov (United States)

    Om, Azlin Che; Ghazali, Amir Hamzah Ahmad; Keng, Chan Lai; Ishak, Zamzuri

    2009-12-01

    Introduction of diazotrophic rhizobacteria to oil palm tissues during the in vitro micropropagation process establishes an early associative interaction between the plant cells and bacteria. In the association, the diazotrophs provide the host plants with phytohormones and fixed nitrogen. This study was conducted to observe growth of bacterised tissue cultured oil palm plants under ex vitro conditions after 280 days of growth. Root dry weight, shoot dry weight, root volume, bacterial colonisation, leaf protein and chlorophyll content of the host plants were observed. The results revealed that the inocula successfully colonised roots of the host plants. Plants inoculated with Acetobacter diazotrophicus (R12) had more root dry weight and volume than plants inoculated with Azospirillum brasilense (Sp7). Leaf protein and chlorophyll content were higher in the bacterised plants compared to Control 2 plants (inoculated with killed Sp7). These results suggest that the diazotrophs successfully improved the growth of the host plant (oil palm) and minimised the amount of N fertiliser necessary for growth.

  1. Changes in ruminal bacterial community composition following feeding of alfalfa silage inoculated with a commercial silage inoculant

    Science.gov (United States)

    Some silage inoculants promote an increase in milk production, possibly through altering the rumen microflora. In this study, dairy cows fed alfalfa silage treated with the inoculant, Lactobacillus plantarum MTD/1 (LPS), were compared to cows fed untreated silage (Ctrl) with the objectives: 1) to de...

  2. A pliable electroporation patch (ep-Patch) for efficient delivery of nucleic acid molecules into animal tissues with irregular surface shapes.

    Science.gov (United States)

    Wei, Zewen; Huang, Yuanyu; Zhao, Deyao; Hu, Zhiyuan; Li, Zhihong; Liang, Zicai

    2015-01-05

    Delivery of nucleic acids into animal tissues by electroporation is an appealing approach for various types of gene therapy, but efficiency of existing methodsis not satisfactory. Here we present the validation of novel electroporation patch (ep-Patch) for efficient delivery of DNA and siRNA into mouse tissues. Using micromachining technology, closely spaced gold electrodes were made on the pliable parylene substrate to form a patch-like electroporation metrics. It enabled large coverage of the target tissues and close surface contact between the tissues and electrodes, thus providing a uniform electric field to deliver nucleic acids into tissues, even beneath intact skin. Using this ep-Patch for efficiently delivery of both DNA and siRNA, non-invasive electroporation of healthy mouse muscle tissue was successfully achieved. Delivery of these nucleic acids was performed to intact tumors with satisfactory results. Silencing of tumor genes using the ep-Patch was also demonstrated on mice. This pliable electroporation patch method constitutes a novel way of in vivo delivery of siRNA and DNA to certain tissues or organs to circumvent the disadvantages of existing methodologies for in vivo delivery of nucleic acid molecules.

  3. Growth and {sup 137}Cs uptake of four Brassica species influenced by inoculation with a plant growth-promoting rhizobacterium Bacillus pumilus in three contaminated farmlands in Fukushima prefecture, Japan

    Energy Technology Data Exchange (ETDEWEB)

    Aung, Han Phyo [United Graduate School of Agricultural Science, Tokyo University of Agriculture and Technology, Saiwaicho 3-5-8, Fuchu, Tokyo 183-8509 (Japan); Djedidi, Salem; Oo, Aung Zaw [Institute of Agriculture, Tokyo University of Agriculture and Technology, Saiwaicho 3-5-8, Fuchu, Tokyo 183-8509 (Japan); Aye, Yi Swe [Department of International Environmental and Agricultural Science, Graduate School of Agriculture, Tokyo University of Agriculture and Technology, Saiwaicho 3-5-8, Fuchu, Tokyo 183-8509 (Japan); Yokoyama, Tadashi; Suzuki, Sohzoh [Institute of Agriculture, Tokyo University of Agriculture and Technology, Saiwaicho 3-5-8, Fuchu, Tokyo 183-8509 (Japan); Sekimoto, Hitoshi [Faculty of Agriculture, Utsunomiya University, 321-8505 (Japan); Bellingrath-Kimura, Sonoko Dorothea, E-mail: skimura@cc.tuat.ac.jp [Institute of Agriculture, Tokyo University of Agriculture and Technology, Saiwaicho 3-5-8, Fuchu, Tokyo 183-8509 (Japan)

    2015-07-15

    The effectiveness of the plant growth-promoting rhizobacterium Bacillus pumilus regarding growth promotion and radiocesium ({sup 137}Cs) uptake was evaluated in four Brassica species grown on different {sup 137}Cs contaminated farmlands at Fukushima prefecture in Japan from June to August 2012. B. pumilus inoculation did not enhance growth in any of the plants, although it resulted in a significant increase of {sup 137}Cs concentration and higher {sup 137}Cs transfer from the soil to plants. The Brassica species exhibited different {sup 137}Cs uptake abilities in the order Komatsuna > turnip > mustard > radish. TF values of {sup 137}Cs ranged from 0.018 to 0.069 for all vegetables. Komatsuna possessed the largest root surface area and root volume, and showed a higher {sup 137}Cs concentration in plant tissue and higher {sup 137}Cs TF values (0.060) than the other vegetables. Higher {sup 137}Cs transfer to plants was prominent in soil with a high amount of organic matter and an Al-vermiculite clay mineral type. - Highlights: • PGPR inoculation did not enhance plant biomass of tested plants. • PGPR inoculation resulted in higher {sup 137}Cs concentration in plants. • Komatsuna that had larger root volume showed higher {sup 137}Cs TF from soil to plants. • Soil with high SOM and Al-vermiculite caused larger {sup 137}Cs transfer to plants.

  4. The influence of surface roughness on the bond strength of composite to dental hard tissues after Er:YAG laser irradiation

    Science.gov (United States)

    Gardner, Andrew K.; Staninec, Michal; Fried, Daniel

    2005-03-01

    The uniformity of laser treated hard tissue surfaces depends on the laser beam quality and the degree of spatial overlap between adjacent laser pulses. Since the surface roughness or surface topography is expected to influence adhesion, our aim in this study was to assess the influence of the surface topography on the adhesion of composite to both enamel and dentin treated at the optimal conditions for the efficient ablation of those tissues with the Er:YAG laser. Human dentin and bovine enamel samples were uniformly irradiated by an Er:YAG laser operating with a pulse duration of 20-30-μs. The laser pulses were 300-μm in diameter with the laser operating in a single TEM00 transverse mode, and the distance between laser spots was varied from 50-200-μm. A motion control system and a pressurized spray system incorporating a microprocessor controlled pulsed nozzle for water delivery, were used to ensure uniform treatment of the entire surface. Shear bond testing was used to evaluate the adhesive strength in order to assess the suitability of laser treated surfaces for bonding. The effect of the degree of overlap of adjacent laser pulses on the surface roughness and the shear bond strength of composite to enamel and dentin is reported.

  5. Mycorrhizal inoculation affects the phytochemical content in strawberry fruits

    Directory of Open Access Journals (Sweden)

    Ana Paula Cecatto

    2016-04-01

    Full Text Available The aim of this research was to evaluate the effect of the inoculation date of arbuscular mycorrhizal fungi on the fruit quality and the content of phytochemicals in a strawberry soilless growing system. The experiment was performed in Huelva (Spain and was conducted in a greenhouse on the La Rábida Campus of Huelva University under natural light and temperature from October 2013 to June 2014. Three short-day strawberry cultivars (‘Splendor’, ‘Sabrina’ and ‘Fortuna’ were grown in polyethylene bags filled with coconut fibres. Randomized block design, with 3 repetitions and factorial arrangement (3 cultivars x 3 treatments, was established. Each replicate consisted of one bag with 12 plants supporting structures at 40 cm height. The treatments were: T1 = mycorrhizal inoculation in the transplantation; T2 = mycorrhizal inoculation 30 days after transplantation (DAT; and T0 = control treatment, without inoculation. Arbuscular mycorrhizal fungi inoculation significantly affected the contents of anthocyanin and phenolics. When the inoculation is performed in the transplantation, the fruits showed a high content of anthocyanin and total phenolics. The mycorrhizal inoculation influences decreasing the acidity in fruit throughout the growing season and increase firmness only during the early stage of production.

  6. Inoculation message treatments for curbing noncommunicable disease development.

    Science.gov (United States)

    Mason, Alicia M; Miller, Claude H

    2013-07-01

    To study the effect of various types of inoculation message treatments on resistance to persuasive and potentially deceptive health- and nutrition-related (HNR) content claims of commercial food advertisers. A three-phase experiment was conducted among 145 students from a Midwestern U.S. university. Quantitative statistical analyses were used to interpret the results. RESULTS provide clear evidence that integrating regulatory focus/fit considerations enhances the treatment effectiveness of inoculation messages. Inoculation messages that employed a preventative, outcome focus with concrete language were most effective at countering HNR advertising claims. The findings indicate that inoculation fosters resistance equally across the most common types of commercially advertised HNR product claims (e.g., absolute, general, and structure/function claims). As the drive to refine the inoculation process model continues, further testing and application of this strategy in a public health context is needed to counter ongoing efforts by commercial food advertisers to avoid government regulations against deceptive practices such as dubious health/nutrition claims. This research advances inoculation theory by providing evidence that 1) good regulatory fit strengthens the effect of refutational preemption and 2) an inoculation approach is highly effective at fostering resistance to commercial advertisers' HNR content claims. This macro approach appears far superior to education or information-based promotional health campaigns targeted solely at specific populations demonstrating rising rates of noncommunicable disease.

  7. Development and evaluation of a new 3-D digitization and computer graphic system to study the anatomic tissue and restoration surfaces.

    Science.gov (United States)

    Dastane, A; Vaidyanathan, T K; Vaidyanathan, J; Mehra, R; Hesby, R

    1996-01-01

    It is necessary to visualize and reconstruct tissue anatomic surfaces accurately for a variety of oral rehabilitation applications such as surface wear characterization and automated fabrication of dental restorations, accuracy of reproduction of impression and die materials, etc. In this investigation, a 3-D digitization and computer-graphic system was developed for surface characterization. The hardware consists of a profiler assembly for digitization in an MTS biomechanical test system with an artificial mouth, an IBM PS/2 computer model 70 for data processing and a Hewlett-Packard laser printer for hardcopy outputs. The software used includes a commercially available Surfer 3-D graphics package, a public domain data-fitting alignment software and an inhouse Pascal program for intercommunication plus some other limited tasks. Surfaces were digitized before and after rotation by angular displacement, the digital data were interpolated by Surfer to provide a data grid and the surfaces were computer graphically reconstructed: Misaligned surfaces were aligned by the data-fitting alignment software under different choices of parameters. The effect of different interpolation parameters (e.g. grid size, method of interpolation) and extent of rotation on the alignment accuracy was determined. The results indicate that improved alignment accuracy results from optimization of interpolation parameters and minimization of the initial misorientation between the digitized surfaces. The method provides important advantages for surface reconstruction and visualization, such as overlay of sequentially generated surfaces and accurate alignment of pairs of surfaces with small misalignment.

  8. Application of a Liquid Extraction Based Sealing Surface Sampling Probe for Mass Spectrometric Analysis of Dried Blood Spots and Mouse Whole-Body Thin Tissue Sections

    Energy Technology Data Exchange (ETDEWEB)

    Van Berkel, Gary J [ORNL; Kertesz, Vilmos [ORNL

    2009-01-01

    The utility of a liquid extraction based sealing surface sampling probe (SSSP) for the direct mass spectrometric analysis of targeted drugs and metabolites in dried blood spots (DBSs) and whole mouse thin tissue sections was demonstrated. The accuracy and precision for the quantitative analysis of a minimum of 50 ng/mL sitamaquine or acetaminophen in DBSs on paper were well within the required 15% dictated by internationally recognized acceptance criteria for assay validations. Analysis of whole-body mouse thin tissue sections from animals dosed with propranolol, adhered to an adhesive tape substrate, provided semi-quantitative information for propranolol and its hydroxyproranolol glucuronide metabolite within specific organs of the tissue. The relative abundances recorded for the two compounds in the brain, lung, kidney and liver were in nominal agreement with previously reported amounts based on analysis using a liquid microjunction surface sampling probe (LMJ-SSP), and whole-body autoradiography (WBA) and HPLC-MS analysis. The ability to sample and analyze from tape-adhered tissue samples, which are generally employed in WBA analysis, presents the possibility of consecutive WBA and SSSP-MS analysis of the same tissue section. This would facilitate assignment, and possibly quantitation, of the different molecular forms of total drug related material detected in the WBA analysis. The flexibility to sample larger or smaller spot sizes, alternative probe sealing mechanisms, and a reduction in internal volumes and associated sample carryover issues will be among the first simple improvements necessary to make the SSSP-MS method a practical DBS and/or thin tissue section analysis tool or to expand its use to other surface sampling applications.

  9. Virological and serological findings in Rousettus aegyptiacus experimentally inoculated with vero cells-adapted hogan strain of Marburg virus.

    Directory of Open Access Journals (Sweden)

    Janusz T Paweska

    Full Text Available The Egyptian fruit bat, Rousettus aegyptiacus, is currently regarded as a potential reservoir host for Marburg virus (MARV. However, the modes of transmission, the level of viral replication, tissue tropism and viral shedding pattern remains to be described. Captive-bred R. aegyptiacus, including adult males, females and pups were exposed to MARV by different inoculation routes. Blood, tissues, feces and urine from 9 bats inoculated by combination of nasal and oral routes were all negative for the virus and ELISA IgG antibody could not be demonstrated for up to 21 days post inoculation (p.i.. In 21 bats inoculated by a combination of intraperitoneal/subcutaneous route, viremia and the presence of MARV in different tissues was detected on days 2-9 p.i., and IgG antibody on days 9-21 p.i. In 3 bats inoculated subcutaneously, viremia was detected on days 5 and 8 (termination of experiment, with virus isolation from different organs. MARV could not be detected in urine, feces or oral swabs in any of the 3 experimental groups. However, it was detected in tissues which might contribute to horizontal or vertical transmission, e.g. lung, intestines, kidney, bladder, salivary glands, and female reproductive tract. Viremia lasting at least 5 days could also facilitate MARV mechanical transmission by blood sucking arthropods and infections of susceptible vertebrate hosts by direct contact with infected blood. All bats were clinically normal and no gross pathology was identified on post mortem examination. This work confirms the susceptibility of R. aegyptiacus to infection with MARV irrespective of sex and age and contributes to establishing a bat-filovirus experimental model. Further studies are required to uncover the mode of MARV transmission, and to investigate the putative role of R. aegyptiacus as a reservoir host.

  10. Immune response of calves inoculated with proteins ofAnaplasma marginale bound to an immunostimulant complex

    Directory of Open Access Journals (Sweden)

    Marcela Ribeiro Gasparini

    Full Text Available Despite our current knowledge of the immunology, pathology, and genetics of Anaplasma marginale, prevention in cattle is currently based on old standbys, including live attenuated vaccines, antibiotic treatment, and maintaining enzootic stability in cattle herds. In the present study, we evaluated the use of an immunostimulant complex (ISCOMATRIX adjuvant, associated with a pool of recombinant major surface proteins (rMSP1a, rMSP1b, rMSP4 and rMSP5 to improve the humoral immune response triggered in calves mainly by IgG2. Ten calves were divided in three groups: 4 calves were inoculated with the ISCOMATRIX/rMSPs (G1; 2 calves were inoculated with ISCOMATRIX adjuvant (G2; and 4 calves received saline (G3. Three inoculations were administered at 21-day intervals. In G1, the calves showed significant increases in total IgG, IgG1 and IgG2 levels 21 days after the second inoculation, compared to the control group (p < 0.05, and G1 calves remained above the cut-off value 28 days after the third inoculation (p < 0.05. The post-immunized sera from calves in G1 reacted specifically for each of the rMSPs used. In conclusion, the ISCOMATRIX/rMSPs induced antigen-specific seroconversion in calves. Therefore, additional testing to explore the protection induced by rMSPs, both alone and in conjunction with proteins previously identified as subdominant epitopes, is warranted.

  11. Neospora caninum NC-6 Argentina induces fetopathy in both serologically positive and negative experimentally inoculated pregnant dams.

    Science.gov (United States)

    Bacigalupe, D; Basso, W; Caspe, S G; Moré, G; Lischinsky, L; Gos, M L; Leunda, M; Campero, L; Moore, D P; Schares, G; Campero, C M; Venturini, M C

    2013-07-01

    Neospora caninum infection is a major cause of abortion in cattle. The objectives of this study were to genetically characterize the N. caninum NC-6 Argentina isolate using a multilocus microsatellite analysis approach and to study its biological behavior by experimental inoculations into seronegative and seropositive pregnant cattle, evaluating the humoral and cellular immune response elicited and the occurrence of transplacental transmission and fetopathy. Pregnant cows (65 days of gestation) seropositive and seronegative to N. caninum were intravenously inoculated with tachyzoites of the NC-6 Argentina N. caninum strain and slaughtered at 108 ± 2 days of gestation. Serum samples were analyzed for N. caninum antibodies by indirect fluorescent antibody test. The cellular immune response was analyzed by detection of gamma interferon (γIFN) production in blood cells. Tissue samples from dams, fetuses, and placental cotyledons were processed by histopathological and immunohistochemical techniques and examined for N. caninum DNA by PCR. Positive DNA samples were further analyzed by multilocus microsatellite typing for N. caninum. Inoculated animals had significantly higher N. caninum antibody titers and γIFN production than control animals. One seropositive inoculated cow aborted, one seronegative cow had a non-viable fetus, and the remaining fetuses from the experimentally inoculated dams had histopathologic lesions. The PCR was positive in 3/4 fetuses from seronegative inoculated cows and in 2/3 fetuses from seropositive inoculated cows. Multilocus microsatellite analysis revealed that the N. caninum DNA present in fetuses and placentas had an identical pattern to NC-6 Argentina strain. The NC-6 Argentina strain proved to be able to cross the placenta and to induce fetopathy in both the seropositive and seronegative dams.

  12. Plasma-induced polymerization as a tool for surface functionalization of polymer scaffolds for bone tissue engineering: an in vitro study.

    Science.gov (United States)

    López-Pérez, Paula M; da Silva, Ricardo M P; Sousa, Rui A; Pashkuleva, Iva; Reis, Rui L

    2010-09-01

    A commonly applied strategy in the field of tissue engineering (TE) is the use of temporary three-dimensional scaffolds for supporting and guiding tissue formation in various in vitro strategies and in vivo regeneration approaches. The interactions of these scaffolds with highly sensitive bioentities such as living cells and tissues primarily occur through the material surface. Hence, surface chemistry and topological features have principal roles in coordinating biological events at the molecular, cellular and tissue levels on timescales ranging from seconds to weeks. However, tailoring the surface properties of scaffolds with a complex shape and architecture remains a challenge in materials science. Commonly applied wet chemical treatments often involve the use of toxic solvents whose oddments in the construct could be fatal in the subsequent application. Aiming to shorten the culture time in vitro (i.e. prior the implantation of the construct), in this work we propose a modification of previously described bone TE scaffolds made from a blend of starch with polycaprolactone (SPCL). The modification method involves surface grafting of sulfonic or phosphonic groups via plasma-induced polymerization of vinyl sulfonic and vinyl phosphonic acid, respectively. We demonstrate herein that the presence of these anionic functional groups can modulate cell adhesion mediated through the adsorbed proteins (from the culture medium). Under the conditions studied, both vitronectin adsorption and osteoblast proliferation and viability increased in the order SPCL plasma-induced polymerization is an excellent alternative route, when compared to the commonly used wet chemical treatments, for the surface functionalization of biodevices with complex shape and porosity.

  13. Effects of surface hydrophilicity and microtopography on early stages of soft and hard tissue integration at non-submerged titanium implants: an immunohistochemical study in dogs.

    Science.gov (United States)

    Schwarz, Frank; Ferrari, Daniel; Herten, Monika; Mihatovic, Ilja; Wieland, Marco; Sager, Martin; Becker, Jürgen

    2007-11-01

    The aim of the present study was to investigate the effects of surface hydrophilicity and microtopography on soft and hard tissue integration at non-submerged titanium implants. Implantation of conventional sand-blasted large grit and acid-etched (SLA) and chemically modified SLA (modSLA) titanium implants with differently structured transmucosal surfaces (SLA implants: machined [M-SLA] or SLA [SLA-SLA]; modSLA implants: mod acid-etched [modA] [modA-modSLA] or modSLA [modSLA-modSLA]) was performed bilaterally in the upper and lower jaws of 15 beagle dogs. The animals were sacrificed after 1, 4, 7, 14, or 28 days. Tissue reactions were assessed histomorphometrically and immunohistochemically using monoclonal antibodies to transglutaminase II (angiogenesis) and osteocalcin. Although the junctional epithelium commonly was separated from M-SLA and SLA-SLA implants by a gap, the epithelial cells appeared to be in close contact with modA-modSLA surfaces after 14 days of healing. Moreover, modA-modSLA and modSLA-modSLA groups showed a well-vascularized subepithelial connective tissue exhibiting collagen fibers that started to extend and attach partially perpendicular to the implant surface. The highest and statistically significant mean bone-to-implant contact areas were observed in the modA-modSLA and modSLA-modSLA groups at days 7, 14, and 28. Within the limits of this study, it may be concluded that soft and hard tissue integration was influenced mainly by surface hydrophilicity rather than by microtopography.

  14. Highly roughened polycaprolactone surfaces using oxygen plasma-etching and in vitro mineralization for bone tissue regeneration: fabrication, characterization, and cellular activities.

    Science.gov (United States)

    Kim, YongBok; Kim, GeunHyung

    2015-01-01

    Herein, poly(ɛ-caprolactone) (PCL) surfaces were treated to form various roughness values (R(a)=290-445 nm) and polar functional groups on the surfaces using a plasma-etching process, followed by immersion into simulated body fluid (SBF) for apatite formation. The surface morphology, chemical composition, and mean roughness of the plasma-etched PCL surfaces were measured, and various physical and morphological properties (water contact angles, protein absorption ability, and crystallite size of the apatite layer) of the in vitro mineralized PCL surfaces were evaluated. The roughened PCL surface P-3, which was treated with a sufficient plasma exposure time (4 h), achieved homogeneously distributed apatite formation after soaking in SBF for 7 days, as compared with other surfaces that were untreated or plasma-treated for 30 min or 2 h. Furthermore, to demonstrate their feasibility as a biomimetic surface, pre-osteoblast cells (MC3T3-E1) were cultured on the mineralized PCL surfaces, and cell viability, DAPI-phalloidin fluorescence assay, and alizarin red-staining of the P-3 surface were highly improved compared to the P-1 surface treated with a 30-min plasma exposure time; compared to untreated mineralized PCL surface (N-P), P-3 showed even greater improvements in cell viability and DAPI-phalloidin fluorescence assay. Based on these results, we found that the mineralized PCL surface supplemented with the appropriate plasma treatment can be implicitly helpful to achieve rapid hard tissue regeneration. Copyright © 2014 Elsevier B.V. All rights reserved.

  15. WE-AB-303-04: A Tissue Model of Cherenkov Emission From the Skin Surface During Megavoltage X-Ray Radiotherapy

    Energy Technology Data Exchange (ETDEWEB)

    Wiles, A. N.; Loyalka, S. K. [University of Missouri, Columbia, MO (United States); Izaguirre, E. W. [University of Missouri, Columbia, MO (United States); Baylor Scott & White Health, Temple, TX (United States)

    2015-06-15

    Purpose: To develop a tissue model of Cherenkov radiation emitted from the skin surface during external beam radiotherapy. Imaging Cherenkov radiation emitted from human skin allows visualization of the beam position and potentially surface dose estimates, and our goal is to characterize the optical properties of these emissions. Methods: We developed a Monte Carlo model of Cherenkov radiation generated in a semi-infinite tissue slab by megavoltage x-ray beams with optical transmission properties determined by a two-layered skin model. We separate the skin into a dermal and an epidermal layer in our model, where distinct molecular absorbers modify the Cherenkov intensity spectrum in each layer while we approximate the scattering properties with Mie and Rayleigh scattering from the highly structured molecular organization found in human skin. Results: We report on the estimated distributions of the Cherenkov wavelength spectrum, emission angles, and surface distribution for the modeled irradiated skin surface. The expected intensity distribution of Cherenkov radiation emitted from skin shows a distinct intensity peak around 475 nm, the blue region of the visible spectrum, between a pair of optical absorption bands in hemoglobin and a broad plateau beginning near 600 nm and extending to at least 700 nm where melanin and hemoglobin absorption are both low. We also find that the Cherenkov intensity decreases with increasing angle from the surface normal, the majority being emitted within 20 degrees of the surface normal. Conclusion: Our estimate of the spectral distribution of Cherenkov radiation emitted from skin indicates an advantage to using imaging devices with long wavelength spectral responsivity. We also expect the most efficient imaging to be near the surface normal where the intensity is greatest; although for contoured surfaces, the relative intensity across the surface may appear to vary due to decreasing Cherenkov intensity with increased angle from the

  16. Inoculation of maize with Azospirillum brasilense in the seed furrow

    OpenAIRE

    Tâmara Prado de Morais; Césio Humberto de Brito; Afonso Maria Brandão; Wender Santos Rezende

    2016-01-01

    ABSTRACT Several studies addressing the inoculation of cereals with diazotrophic microorganisms can be found in the literature. However, in many experiments, investigators have overlooked the feasibility of applying these microorganisms to the furrow together with the seed, and the effect of bacterial concentration on phytostimulation. The aim of this work was to evaluate the effect of doses of an inoculant based on Azospirillum brasilense, applied to the seed furrow when planting maize, comb...

  17. Albendazole inhibits Pneumocystis carinii proliferation in inoculated immunosuppressed mice.

    OpenAIRE

    Bartlett, M S; Edlind, T D; Lee, C. H.; Dean, R; Queener, S F; Shaw, M M; Smith, J W.

    1994-01-01

    Albendazole, a benzimidazole derivative widely used for treating helminth infections, was successfully used to treat and prevent development of Pneumocystis carinii pneumonia in transtracheally inoculated immunosuppressed mice. For treatment, 3 weeks postinoculation, albendazole at 300 and 600 mg/kg of body weight per day was administered in food for 3 weeks. For prophylaxis, albendazole was begun on the same day as inoculation at 300 mg/kg/day for 7 days, and then the dose was reduced to 150...

  18. Effect of inoculating addition on machinability of gray cast iron

    Institute of Scientific and Technical Information of China (English)

    REN Fengzhang; LI Fengjun; LIU Weiming; MA Zhanhong; TIAN Baohong

    2009-01-01

    Gray cast irons were inoculated with FeSi75+RE and FeSi75+Sr inoculants. The changes of apex angle of the drills before and af-ter being used were used to evaluate machinability of gray cast irons. Effect of FeSi75+RE and FeSi75+Sr inoculants on mechanical proper-ties, machinability and sensibility of gray cast iron used in cylinder block were investigated. Experimental results showed that gray cast iron treated with 60%FeSi75+40% RE inoculants exhibited tensile strength consistently at about 295 MPa along with good hardness and im-proved metallurgical quality. While gray cast iron inoculated with 20%FeSi75+80% Sr inoculants exhibited the best machinability, the low-est cross-section sensibility and the least microhardness difference. The tool flank wear of the drill increased correspondingly with the in-crease of the microhardness difference of the matrix, indicating the great effect of homogeneity of the matrix on the machinability of gray cast iron.

  19. Conditions Affecting Shelf-Life of Inoculated Legume Seed

    Directory of Open Access Journals (Sweden)

    Greg Gemell

    2012-02-01

    Full Text Available Microbial inoculants are becoming more available as sustainable alternatives to fertilizers and other agrichemicals in broad-acre cropping. However, with the exception of legume inoculants little is understood about effective delivery and survival of the inoculum. Legume inoculants are applied to both seed and soil but seed inoculation is the most economical technique. Large quantities of pasture seed in Australia are inoculated by commercial seed coating companies, but the long-term survival of seed-applied inoculum is variable and monitoring of viability requires specialist microbiology skills and facilities. The aim of our research was to define optimum storage conditions for survival of rhizobia on legume seed and evaluate water activity as a means of monitoring shelf-life. The relationship between survival and water activity varied according to seed species, inoculum preparation, coating ingredients, initial water activity and time suggesting that storage conditions would need to be defined for each different combination. Although drying seeds after coating significantly reduced viable numbers of rhizobia, survival of rhizobia on dried commercially coated lucerne seed after 11 weeks was less variable than seeds that had not been dried. The highest numbers were maintained when seeds remained dry with water activities of between 0.47 and 0.38. The quality of inoculated seed could be improved by reducing the death rate of inoculum during preparation and providing optimum storage conditions for long-term survival.

  20. Surface modification of zirconia with polydopamine to enhance fibroblast response and decrease bacterial activity in vitro: A potential technique for soft tissue engineering applications.

    Science.gov (United States)

    Liu, Mingyue; Zhou, Jianfeng; Yang, Yang; Zheng, Miao; Yang, Jianjun; Tan, Jianguo

    2015-12-01

    The quality of soft-tissue integration plays an important role in the short- and long-term success of dental implants. The aim of the present study was to provide a surface modification approach for zirconia implant abutment materials and to evaluate its influence on fibroblast behavior and oral bacteria adhesion, which are the two main factors influencing the quality of peri-implant soft-tissue seal. In this study, polydopamine (PDA)-coated zirconia was prepared and the surface characteristics were evaluated using scanning electron microscopy, atomic force microscopy, a contact-angle-measuring device, X-ray photoelectron spectroscopy, and Raman spectroscopy. The responses of human gingival fibroblasts (HGFs) to PDA-coated zirconia; i.e., adhesion, proliferation, morphology, protein synthesis, and gene expression, were analyzed. Additionally, the adhesion of Streptococcus gordonii and Streptococcus mutans to zirconia after PDA coating was assessed by scanning electron microscopy and live/dead staining. The material surface analyses suggested the successful coating of PDA onto the zirconia surface. The PDA coating significantly increased cell adhesion and proliferation compared with pristine zirconia. HGFs exhibited a high degree of spreading and secreted a high level of collagen type I on PDA-modified disks. Upregulation of integrin α5, β1, β3 and fibronectin was noted in HGFs cultured on PDA-coated zirconia. The number of adherent bacteria decreased significantly on zirconia after PDA coating. In summary, our result suggest that PDA is able to modify the surface of zirconia, influence HGFs' behavior and reduce bacterial adhesion. Therefore, this surface modification approach holds great potential for improving soft-tissue integration around zirconia abutments in clinical application.

  1. Biochemical Response of Glycine Max after Inoculation of Different Concentrations of Fusarium Solani

    Directory of Open Access Journals (Sweden)

    N. Hamid

    2014-06-01

    Full Text Available Diseases of agricultural crops are old as agriculture itself. It has been estimated that fungal diseases are responsible for an annual reduction for more than 20% of total potential world food production from crop plants. Randomized complete block design experiments were conducted to study the effect of different colonies of Fusarium solani on some biochemical and physiological parameters of Glycine max. Sterilized seeds of Glycine max (L. Merr was artificially inoculated with conidia of 3 different concentrations (10,000, 100,000 and 1000, 000 cfu. Seedlings of Glycine max was grown in 350g acid washed sand. Complete Nutrient Hoagland solution was regularly used to irrigate the plants. Leaves samples were weekly collected for analysis of biochemical tests. It was observed that inoculation of Fusarium solani significantly increase the concentration of total soluble carbohydrate and reducing sugar in inoculated plants over healthy tissues. Protein analysis revealed a significant reduction with the development of disease. Activity of invertase illustrate a variable change initially it was increased than afterward declined.

  2. Gene expression changes in Porphyromonas gingivalis W83 after inoculation in rat oral cavity.

    Science.gov (United States)

    Zhao, Jian; Li, Qian; Pan, Chun-Ling; Liu, Jun-Chao; Wang, Hong-Yan; Tan, Li-Si; Pan, Ya-Ping

    2015-05-24

    The development of chronic periodontitis was due to not only periodontal pathogens, but also the interaction between periodontal pathogens and host. The aim of this study is to investigate the alterations in gene expression in Porphyromonas gingivalis (P.gingivalis) W83 after inoculation in rat oral cavity. P.gingivalis W83 inoculation in rat oral cavity caused inflammatory responses in gingival tissues and destroyed host alveolar bone. Microarray analysis revealed that 42 genes were upregulated, and 22 genes were downregulated in the detected 1786 genes in the inoculated P.gingivalis W83. Real-time quantitative PCR detection confirmed the expression alterations in some selected genes. Products of these upregulated and downregulated genes are mainly related to transposon functions, cell transmembrane transportation, protein and nucleic acid metabolism, energy metabolism, cell division and bacterial pathogenicity. P.gingivalis W83 has a pathogenic effect on host oral cavity. Meanwhile, inflammatory oral environment alters P.gingivalis W83 gene expression profile. These changes in gene expression may limit the proliferation and weaken the pathogenicity of P.gingivalis W83, and favor themselves to adapt local environment for survival.

  3. Order of inoculation affects the success of co-invading entomopathogenic fungi.

    Science.gov (United States)

    Zamora-Macorra, E J; Guzmán-Franco, A W; Pell, J K; Alatorre-Rosas, R; Suarez-Espinoza, J

    2012-12-01

    The effect of order of inoculation of Pandora blunckii and Zoophthora radicans co-infecting Plutella xylostella (L.) (Lepidoptera: Plutellidae) was investigated. After co-inoculation, the proportion of larvae infected by either species was greatly reduced compared to when they were inoculated singly. The order of inoculation influenced the final outcome; the isolate inoculated last always killed more larvae than the isolate inoculated first.

  4. Effects of surface modification on the mechanical and structural properties of nanofibrous poly(ε-caprolactone)/forsterite scaffold for tissue engineering applications.

    Science.gov (United States)

    Kharaziha, M; Fathi, M H; Edris, H

    2013-12-01

    Composite scaffolds consisting of polymers reinforced with ceramic nanoparticles are widely applied for hard tissue engineering. However, due to the incompatible polarity of ceramic nanoparticles with polymers, they tend to agglomerate in the polymer matrix which results in undesirable effects on the integral properties of composites. In this research, forsterite (Mg2SiO4) nanoparticles was surface esterified by dodecyl alcohol and nanofibrous poly(ε-caprolactone)(PCL)/modified forsterite scaffolds were developed through electrospinning technique. The aim of this research was to investigate the properties of surface modified forsterite nanopowder and PCL/modified forsterite scaffolds, before and after hydrolytic treatment, as well as the cellular attachment and proliferation. Results demonstrated that surface modification of nanoparticles significantly enhanced the tensile strength and toughness of scaffolds upon 1.5- and 4-folds compared to unmodified samples, respectively, due to improved compatibility between matrix and filler. Hydrolytic treatment of scaffolds also modified the bioactivity and cellular attachment and proliferation due to greatly enhanced hydrophilicity of the forsterite nanoparticles after this process compared to surface modified samples. Results suggested that surface modification of forsterite nanopowder and hydrolytic treatment of the developed scaffolds were effective approaches to address the issues in the formation of composite fibers and resulted in development of bioactive composite scaffolds with ideal mechanical and structural properties for bone tissue engineering applications.

  5. Genetic diversity of symbiotic Bradyrhizobium elkanii populations recovered from inoculated and non-inoculated Acacia mangium field trials in Brazil.

    Science.gov (United States)

    Perrineau, M M; Le Roux, C; de Faria, S M; de Carvalho Balieiro, F; Galiana, A; Prin, Y; Béna, G

    2011-07-01

    Acacia mangium is a legume tree native to Australasia. Since the eighties, it has been introduced into many tropical countries, especially in a context of industrial plantations. Many field trials have been set up to test the effects of controlled inoculation with selected symbiotic bacteria versus natural colonization with indigenous strains. In the introduction areas, A. mangium trees spontaneously nodulate with local and often ineffective bacteria. When inoculated, the persistence of inoculants and possible genetic recombination with local strains remain to be explored. The aim of this study was to describe the genetic diversity of bacteria spontaneously nodulating A. mangium in Brazil and to evaluate the persistence of selected strains used as inoculants. Three different sites, several hundred kilometers apart, were studied, with inoculated and non-inoculated plots in two of them. Seventy-nine strains were isolated from nodules and sequenced on three housekeeping genes (glnII, dnaK and recA) and one symbiotic gene (nodA). All but one of the strains belonged to the Bradyrhizobium elkanii species. A single case of housekeeping gene transfer was detected among the 79 strains, suggesting an extremely low rate of recombination within B. elkanii, whereas the nodulation gene nodA was found to be frequently transferred. The fate of the inoculant strains varied depending on the site, with a complete disappearance in one case, and persistence in another. We compared our results with the sister species Bradyrhizobium japonicum, both in terms of population genetics and inoculant strain destiny. Copyright © 2011 Elsevier GmbH. All rights reserved.

  6. Paenibacillus lentimorbus Inoculation Enhances Tobacco Growth and Extenuates the Virulence of Cucumber mosaic virus.

    Directory of Open Access Journals (Sweden)

    Susheel Kumar

    Full Text Available Previous studies with Paenibacillus lentimorbus B-30488" (hereafter referred as B-30488, a plant growth promoting rhizobacteria (PGPR isolated from cow's milk, revealed its capabilities to improve plant quality under normal and stress conditions. Present study investigates its potential as a biocontrol agent against an economically important virus, Cucumber mosaic virus (CMV, in Nicotiana tabacum cv. White Burley plants and delineates the physical, biophysical, biochemical and molecular perturbations due to the trilateral interactions of PGPR-host-CMV. Soil inoculation of B-30488 enhanced the plant vigor while significantly decreased the virulence and virus RNA accumulation by ~12 fold (91% in systemic leaves of CMV infected tobacco plants as compared to the control ones. Histology of these leaves revealed the improved tissue's health and least aging signs in B-30488 inoculated tobacco plants, with or without CMV infection, and showed lesser intercellular spaces between collenchyma cells, reduced amount of xyloglucans and pectins in connecting primary cells, and higher polyphenol accumulation in hypodermis layer extending to collenchyma cells. B-30488 inoculation has favorably maneuvered the essential biophysical (ion leakage and photosynthetic efficiency and biochemical (sugar, proline, chlorophyll, malondialdehyde, acid phosphatase and alkaline phosphatase attributes of tobacco plants to positively regulate and release the virus stress. Moreover, activities of defense related enzymes (ascorbate peroxidase, guaiacol peroxidase, superoxide dismutase and catalase induced due to CMV-infection were ameliorated with inoculation of B-30488, suggesting systemic induced resistance mediated protection against CMV in tobacco. The quantitative RT-PCR analyses of the genes related to normal plant development, stress and pathogenesis also corroborate well with the biochemical data and revealed the regulation (either up or down of these genes in favor of

  7. Microarray expression in peri-implant tissue next to different titanium implant surfaces predicts clinical outcomes: a split-mouth study.

    Science.gov (United States)

    Menini, M; Dellepiane, E; Baldi, D; Longobardi, M G; Pera, P; Izzotti, A

    2017-09-01

    This split-mouth study evaluated miRNA expression of tissues around implants with different surface treatments. Each patient of the sample (five men and five women) received two implants (one control and one test) into an edentulous quadrant to support fixed partial dentures. The control implants (Osseotite) had a dual acid-etched (DAE) surface in the apical portion and a machined coronal part, test implants (Full Osseotite, FOSS) were completely DAE. Machined healing abutments were placed on control implants and DAE abutments on test ones. All implants were assigned codes for blinding. Standardized periapical radiographs were taken at baseline, 2 and 6 months, and 1 year after surgery. Plaque index (PI), bleeding on probing (BOP), and probing depth (PD) were recorded at 3 and 6 weeks, and 2, 3, 6, and 12 months post-implant placement. After 3 months, a mini-invasive sample of soft tissue was collected from seven patients (four women and three men) for miRNA microarray analysis. Control implants showed greater bone resorption (BR) and lower PI: this was not statistically significant. No statistically significant differences in BOP and PD appeared. miRNA modulated by implant surfaces as well as by other clinical conditions has been identified. miRNA microarray analysis revealed that: (i) implant sites with low PI and absence of BOP had a miRNA expression profile similar to those with plaque and absence of BOP; sites with high PI and high BOP had a different profile. (ii) Implant sites with BOP presented similar profiles independently from implant surface. (iii) Implant sites with high PI and normal BR differed from others for miRNA expression profile. (iv) Implant sites with normal BR despite high BOP differed from others. This profile resembled that of FOSS implants. (v) Implant surface affected BR; groups having similar BR clusterized differently according to the implant type. DAE surfaces induced lower BR and more plaque accumulation: This did not affect the

  8. Effects of surface modification on the mechanical and structural properties of nanofibrous poly(ε-caprolactone)/forsterite scaffold for tissue engineering applications

    Energy Technology Data Exchange (ETDEWEB)

    Kharaziha, M., E-mail: Kharaziha.ma@yahoo.com [Biomaterials Research Group, Department of Materials Engineering, Isfahan University of Technology, Isfahan 8415683111 (Iran, Islamic Republic of); Department of Materials Engineering, Isfahan University of Technology, Isfahan 8415683111 (Iran, Islamic Republic of); Fathi, M.H. [Biomaterials Research Group, Department of Materials Engineering, Isfahan University of Technology, Isfahan 8415683111 (Iran, Islamic Republic of); Dental Materials Research Center, Isfahan University of Medical Sciences, Isfahan (Iran, Islamic Republic of); Edris, H. [Biomaterials Research Group, Department of Materials Engineering, Isfahan University of Technology, Isfahan 8415683111 (Iran, Islamic Republic of); Department of Materials Engineering, Isfahan University of Technology, Isfahan 8415683111 (Iran, Islamic Republic of)

    2013-12-01

    Composite scaffolds consisting of polymers reinforced with ceramic nanoparticles are widely applied for hard tissue engineering. However, due to the incompatible polarity of ceramic nanoparticles with polymers, they tend to agglomerate in the polymer matrix which results in undesirable effects on the integral properties of composites. In this research, forsterite (Mg{sub 2}SiO{sub 4}) nanoparticles was surface esterified by dodecyl alcohol and nanofibrous poly(ε-caprolactone)(PCL)/modified forsterite scaffolds were developed through electrospinning technique. The aim of this research was to investigate the properties of surface modified forsterite nanopowder and PCL/modified forsterite scaffolds, before and after hydrolytic treatment, as well as the cellular attachment and proliferation. Results demonstrated that surface modification of nanoparticles significantly enhanced the tensile strength and toughness of scaffolds upon 1.5- and 4-folds compared to unmodified samples, respectively, due to improved compatibility between matrix and filler. Hydrolytic treatment of scaffolds also modified the bioactivity and cellular attachment and proliferation due to greatly enhanced hydrophilicity of the forsterite nanoparticles after this process compared to surface modified samples. Results suggested that surface modification of forsterite nanopowder and hydrolytic treatment of the developed scaffolds were effective approaches to address the issues in the formation of composite fibers and resulted in development of bioactive composite scaffolds with ideal mechanical and structural properties for bone tissue engineering applications. - Highlights: • Forsterite nanopowder was surface modified with dodecyl alcohol. • Nanofibrous PCL/forsterite scaffolds were developed through electrospinning. • Composite scaffolds were treated in boiled water to remove the dodecyl chains. • Surface modification resulted in improved mechanical properties. • Hydrolytic treatment

  9. Cold atmospheric plasma disinfection of cut fruit surfaces contaminated with migrating microorganisms.

    Science.gov (United States)

    Perni, Stefano; Shama, Gilbert; Kong, M G

    2008-08-01

    The efficacy of cold atmospheric gas plasmas against Escherichia coli type 1, Saccharomyces cerevisiae, Gluconobacter liquefaciens, and Listeria monocytogenes Scott A was examined on inoculated membrane filters and inoculated fruit surfaces. Inoculated samples were exposed to a cold atmospheric plasma plume generated by an AC voltage of 8 kV at 30 kHz. The cold atmospheric plasma used in this study was very efficient in reducing the microbial load on the surfaces of filter membranes. However, its efficacy was markedly reduced for microorganisms on the cut surfaces. This lack of effect was not the result of quenching of reactive plasma species responsible for microbial inactivation but principally the result of the migration of microorganisms from the exterior of the fruit tissue to its interior. The velocity of migration through melon tissues was estimated to be around 300 microm min(-1) for E. coli and S. cerevisiae and through mango tissues to be 75 to 150 microm min(-1). These data can serve as operational targets for optimizing the performance of gas plasma inactivation processes. The current capabilities of cold atmospheric plasmas are reviewed and ways to improve their bactericidal efficacy are identified and discussed. Considerable scope exists to enhance significantly the efficacy of cold atmospheric plasmas for decontaminating fresh cut fruits.

  10. Modeling mechanical signals on the surface of µCT and CAD based rapid prototype scaffold models to predict (early stage) tissue development.

    Science.gov (United States)

    Hendrikson, W J; van Blitterswijk, C A; Verdonschot, N; Moroni, L; Rouwkema, J

    2014-09-01

    In the field of tissue engineering, mechano-regulation theories have been applied to help predict tissue development in tissue engineering scaffolds in the past. For this, finite element models (FEMs) were used to predict the distribution of strains within a scaffold. However, the strains reported in these studies are volumetric strains of the material or strains developed in the extracellular matrix occupying the pore space. The initial phase of cell attachment and growth on the biomaterial surface has thus far been neglected. In this study, we present a model that determines the magnitude of biomechanical signals on the biomaterial surface, enabling us to predict cell differentiation stimulus values at this initial stage. Results showed that magnitudes of the 2D strain--termed surface strain--were lower when compared to the 3D volumetric strain or the conventional octahedral shear strain as used in current mechano-regulation theories. Results of both µCT and CAD derived FEMs from the same scaffold were compared. Strain and fluid shear stress distributions, and subsequently the cell differentiation stimulus, were highly dependent on the pore shape. CAD models were not able to capture the distributions seen in the µCT FEM. The calculated mechanical stimuli could be combined with current mechanobiological models resulting in a tool to predict cell differentiation in the initial phase of tissue engineering. Although experimental data is still necessary to properly link mechanical signals to cell behavior in this specific setting, this model is an important step towards optimizing scaffold architecture and/or stimulation regimes. © 2014 Wiley Periodicals, Inc.

  11. Effects of inoculation of biosurfactant-producing Bacillus sp. J119 on plant growth and cadmium uptake in a cadmium-amended soil.

    Science.gov (United States)

    Sheng, Xiafang; He, Linyan; Wang, Qingya; Ye, Hesong; Jiang, Chunyu

    2008-06-30

    A biosurfactant-producing Bacillus sp. J119 isolated from heavy metal contaminated soils was investigated for its effects on the plant growth-promoting characteristics and heavy metal and antibiotic resistance. A pot experiment was conducted for investigating the capability of the biosurfactant-producing bacterial strain Bacillus sp. J119 to promote the plant growth and cadmium uptake of rape, maize, sudangrass and tomato in soil artificially contaminated with different levels of cadmium (Cd) (0 and 50mgkg(-1)). The strain was found to exhibit different multiple heavy metal (Pb, Cd, Cu, Ni and Zn) and antibiotic (kanamycin, streptomycin, ampicillin, tetracycline and rifampin) resistance characteristics. The strain had the capacity to produce indole acetic acid (IAA) and siderophores. Cd treatment did not significantly decreased growth of tomato, maize and rape plants, but Cd treatment significantly decreased growth of sudangrass (p<0.05). In the Cd-added soil, above-ground biomass and root dry weights of tomatoes were increased by 24 and 59%, respectively, in live bacterial inoculation compared to dead bacterial inoculation control. There were no obvious differences in the above-ground tissue and root dry weight of maize and sudangrass between live bacterial inoculation and dead bacterial inoculation. In the soil treated with 50 mg Cdkg(-1), increase in above-ground tissue Cd content varied from 39 to 70% in live bacterium-inoculated plants compared to dead bacterium-inoculated control. In addition, among the inoculated plants, tomato was the greatest Cd accumulator. The bacterial strain was also able to colonize and develop in the rhizosphere soils after root inoculation.

  12. Effects of inoculation of biosurfactant-producing Bacillus sp. J119 on plant growth and cadmium uptake in a cadmium-amended soil

    Energy Technology Data Exchange (ETDEWEB)

    Sheng Xiafang [MOA Key Laboratory of Microbiological Engineering of Agricultural Environment, College of Life Science, Nanjing Agricultural University, Nanjing 210095 (China)], E-mail: xfsheng604@sohu.com; He Linyan; Wang Qingya; Ye Hesong; Jiang Chunyu [MOA Key Laboratory of Microbiological Engineering of Agricultural Environment, College of Life Science, Nanjing Agricultural University, Nanjing 210095 (China)

    2008-06-30

    A biosurfactant-producing Bacillus sp. J119 isolated from heavy metal contaminated soils was investigated for its effects on the plant growth-promoting characteristics and heavy metal and antibiotic resistance. A pot experiment was conducted for investigating the capability of the biosurfactant-producing bacterial strain Bacillus sp. J119 to promote the plant growth and cadmium uptake of rape, maize, sudangrass and tomato in soil artificially contaminated with different levels of cadmium (Cd) (0 and 50 mg kg{sup -1}). The strain was found to exhibit different multiple heavy metal (Pb, Cd, Cu, Ni and Zn) and antibiotic (kanamycin, streptomycin, ampicillin, tetracycline and rifampin) resistance characteristics. The strain had the capacity to produce indole acetic acid (IAA) and siderophores. Cd treatment did not significantly decreased growth of tomato, maize and rape plants, but Cd treatment significantly decreased growth of sudangrass (p < 0.05). In the Cd-added soil, above-ground biomass and root dry weights of tomatoes were increased by 24 and 59%, respectively, in live bacterial inoculation compared to dead bacterial inoculation control. There were no obvious differences in the above-ground tissue and root dry weight of maize and sudangrass between live bacterial inoculation and dead bacterial inoculation. In the soil treated with 50 mg Cd kg{sup -1}, increase in above-ground tissue Cd content varied from 39 to 70% in live bacterium-inoculated plants compared to dead bacterium-inoculated control. In addition, among the inoculated plants, tomato was the greatest Cd accumulator. The bacterial strain was also able to colonize and develop in the rhizosphere soils after root inoculation.

  13. GERMINATION OF GRASSES DUE TO INOCULATION DIAZOTROPHIC BACTERIA

    Directory of Open Access Journals (Sweden)

    C. D. A. Moreira

    2014-07-01

    Full Text Available The germination of forage grasses suffers from numbness and a natural tendency to low quality. The use of microorganisms inoculated in seeds with the purpose of increasing and meet the demand of some nutrient has been shown to be efficient, but the role of the microorganism in germination and rate of force is still unknown. Therefore the goal as study was to evaluate the germination rate of seeds of three cultivars of Brachiaria brizantha CV. Marandu, b., b. brizantha CV. Xaraés and b. humidícola cv Tupi and a cultivar of millet, P. hybrid cv Massai depending on the bacterium Azospirillum brasilense diazotrofic inoculation (nitrogen-fixing. Germination test was used in seed dispersal to assess the effect of first count (VPC in the treatments with and without inoculation. It was done also conducted further tests of electrical conductivity, weight of thousand seeds and water content. The delineation used was randomized entirely (DIC and the statistical analysis carried out through the analysis of variance and comparison of means using the Tukey test, the 5% probability. Massai grass seeds have the highest rate of force of first count in both treatments. Inoculation of bacterium Azospirillum brasilense did not affect the values of force of first count on seeds of the cultivars Marandu, Xaraés, Tupi and Massai. The seeds of the massai have higher germination speed relative the other cultivars evaluated when inoculated.

  14. Plant growth-promoting bacteria as inoculants in agricultural soils.

    Science.gov (United States)

    Souza, Rocheli de; Ambrosini, Adriana; Passaglia, Luciane M P

    2015-12-01

    Plant-microbe interactions in the rhizosphere are the determinants of plant health, productivity and soil fertility. Plant growth-promoting bacteria (PGPB) are bacteria that can enhance plant growth and protect plants from disease and abiotic stresses through a wide variety of mechanisms; those that establish close associations with plants, such as the endophytes, could be more successful in plant growth promotion. Several important bacterial characteristics, such as biological nitrogen fixation, phosphate solubilization, ACC deaminase activity, and production of siderophores and phytohormones, can be assessed as plant growth promotion (PGP) traits. Bacterial inoculants can contribute to increase agronomic efficiency by reducing production costs and environmental pollution, once the use of chemical fertilizers can be reduced or eliminated if the inoculants are efficient. For bacterial inoculants to obtain success in improving plant growth and productivity, several processes involved can influence the efficiency of inoculation, as for example the exudation by plant roots, the bacterial colonization in the roots, and soil health. This review presents an overview of the importance of soil-plant-microbe interactions to the development of efficient inoculants, once PGPB are extensively studied microorganisms, representing a very diverse group of easily accessible beneficial bacteria.

  15. Plant growth-promoting bacteria as inoculants in agricultural soils

    Directory of Open Access Journals (Sweden)

    Rocheli de Souza

    2015-01-01

    Full Text Available AbstractPlant-microbe interactions in the rhizosphere are the determinants of plant health, productivity and soil fertility. Plant growth-promoting bacteria (PGPB are bacteria that can enhance plant growth and protect plants from disease and abiotic stresses through a wide variety of mechanisms; those that establish close associations with plants, such as the endophytes, could be more successful in plant growth promotion. Several important bacterial characteristics, such as biological nitrogen fixation, phosphate solubilization, ACC deaminase activity, and production of siderophores and phytohormones, can be assessed as plant growth promotion (PGP traits. Bacterial inoculants can contribute to increase agronomic efficiency by reducing production costs and environmental pollution, once the use of chemical fertilizers can be reduced or eliminated if the inoculants are efficient. For bacterial inoculants to obtain success in improving plant growth and productivity, several processes involved can influence the efficiency of inoculation, as for example the exudation by plant roots, the bacterial colonization in the roots, and soil health. This review presents an overview of the importance of soil-plant-microbe interactions to the development of efficient inoculants, once PGPB are extensively studied microorganisms, representing a very diverse group of easily accessible beneficial bacteria.

  16. Evaluation of Varying Biochars as Carrier Materials for Bacterial Soil Inoculants

    Science.gov (United States)

    Hale, Lauren; Crowley, David

    2014-05-01

    The incorporation of biochar into agricultural soils for carbon sequestration and improved soil fertility creates an opportunity to simultaneously deliver plant-growth promoting rhizobacteria (PGPR). Many characteristics of biochar materials indicate that these particles could be conducive as inoculum carriers. This could provide a value-added component for biochar marketing and has an advantage over traditional carrier materials, which can be unsustainable or expensive to produce. Here, we assessed the suitability of 10 biochar types, made from 5 feedstocks at 2 pyrolysis temperatures (300°C and 600°C), to serve as carriers for 2 model PGPR strains, Enterobacter cloacae UW5 and Pseudomonas putida UW4. All biochars were characterized based on BET specific surface area, C-N content, pH, EC, and their abilities to adsorb bacterial cells from a liquid inoculum. Further studies incorporated qPCR to quantify the survival of inoculants after introduction into soils via biochar carriers. The biochars that performed well were further assayed for their influence on PGPR traits, 1-aminocyclopropane-1-carboxylate (ACC) deaminase and auxin production. Peat and vermiculite served as traditional carrier materials to which we compared the biochars. Our findings indicated that biochars varied in their interactions with our model PGPR strains. Based on our analysis several biochar types were able to serve as carriers which were as good, if not better than, the traditional carrier materials. Future work should seek to assess shelf life and varying inoculation methods for the biochar-inoculant complexes.

  17. Fabrication of novel high surface area mushroom gilled fibers and their effects on human adipose derived stem cells under pulsatile fluid flow for tissue engineering applications.

    Science.gov (United States)

    Tuin, Stephen A; Pourdeyhimi, Behnam; Loboa, Elizabeth G

    2016-05-01

    The fabrication and characterization of novel high surface area hollow gilled fiber tissue engineering scaffolds via industrially relevant, scalable, repeatable, high speed, and economical nonwoven carding technology is described. Scaffolds were validated as tissue engineering scaffolds using human adipose derived stem cells (hASC) exposed to pulsatile fluid flow (PFF). The effects of fiber morphology on the proliferation and viability of hASC, as well as effects of varied magnitudes of shear stress applied via PFF on the expression of the early osteogenic gene marker runt related transcription factor 2 (RUNX2) were evaluated. Gilled fiber scaffolds led to a significant increase in proliferation of hASC after seven days in static culture, and exhibited fewer dead cells compared to pure PLA round fiber controls. Further, hASC-seeded scaffolds exposed to 3 and 6dyn/cm(2) resulted in significantly increased mRNA expression of RUNX2 after one hour of PFF in the absence of soluble osteogenic induction factors. This is the first study to describe a method for the fabrication of high surface area gilled fibers and scaffolds. The scalable manufacturing process and potential fabrication across multiple nonwoven and woven platforms makes them promising candidates for a variety of applications that require high surface area fibrous materials. We report here for the first time the successful fabrication of novel high surface area gilled fiber scaffolds for tissue engineering applications. Gilled fibers led to a significant increase in proliferation of human adipose derived stem cells after one week in culture, and a greater number of viable cells compared to round fiber controls. Further, in the absence of osteogenic induction factors, gilled fibers led to significantly increased mRNA expression of an early marker for osteogenesis after exposure to pulsatile fluid flow. This is the first study to describe gilled fiber fabrication and their potential for tissue engineering

  18. The use of non-viral gene vectors for bioactive poly-(D,L-lactide implant surfaces in bone tissue engineering

    Directory of Open Access Journals (Sweden)

    AK Reckhenrich

    2012-06-01

    Full Text Available The application of scaffolds in bone tissue engineering often comes along with side effects such as poor integrity, low regeneration rates of bone tissue with inadequate functionality, and, in case of non-degradable implants, the necessity of a second removal surgery after therapy. In this study, we coated a bioresorbable FDA-approved poly-(ε-caprolactone-scaffold for bone regeneration with a poly-(D,L-lactide layer containing copolymer-protected gene vectors to locally provide bone morphogenetic protein-2 (BMP-2. Results show that the presence of such gene vectors did not affect the distribution and attachment of seeded cells on gene-activated surfaces. BMP-2 was released into cell culture supernatants and furthermore detected in homogenised scaffolds. Increased amounts of osteoblastic markers, such as osteocalcin, osteopontin and the activity of alkaline phosphatase, in gene-activated scaffolds in vitro suggest a transdifferentiation of myoblastic C2C12 cells into the osteoblastic phenotype. With this study we present a new technology to bioactivate implant surfaces with non-viral gene vectors. This tool allows the stimulation of tissue regeneration by a local release of therapeutic proteins in vivo.

  19. Magnetically labeled cells with surface-modified fe3 o4 spherical and rod-shaped magnetic nanoparticles for tissue engineering applications.

    Science.gov (United States)

    Gil, Sara; Correia, Clara R; Mano, João F

    2015-04-22

    Magnetically targeted cells with internalized magnetic nanoparticles (MNPs) could allow the success of cell transplantation and cell-based therapies, overcoming low cell retention that occurs when delivering cells by intravenous or local injection. Upon magnetization, these cells could then accumulate and stimulate the regeneration of the tissue in situ. Magnetic targeting of cells requires a detailed knowledge between interactions of engineered nanomaterials and cells, in particular the influence of shape and surface functionalization of MNPs. For the first time, cellular internalization of amino surface-modified iron oxide nanoparticles of two different shapes (nanospheres or nanorods) is studied. MNPs show high cellular uptake and labeled cells could exhibit a strong reaction with external magnetic fields. Compared to nanorods, nanospheres show better internalization efficiency, and labeled cells exhibit strong transportation reaction with external magnetic fields. Contiguous viable cell-sheets are developed by magnetic-force-based tissue engineering. The results confirm that the developed magnetic-responsive nano-biomaterials have potential applicability in tissue engineering or cellular therapies.

  20. Quantitative Phosphoproteomic Analysis of Soybean Root Hairs Inoculated with Bradyrhizobium japonicum

    Energy Technology Data Exchange (ETDEWEB)

    Nguyen, Tran H.; Brechenmacher, Laurent; Aldrich, Joshua T.; Clauss, Therese RW; Gritsenko, Marina A.; Hixson, Kim K.; Libault, Marc; Tanaka, Kiwamu; Yang, Feng; Yao, Qiuming; Pasa-Tolic, Ljiljana; Xu, Dong; Nguyen, Henry T.; Stacey, Gary

    2012-11-11

    Root hairs are single hair-forming cells on roots that function to increase root surface area, enhancing water and nutrient uptake. In leguminous plants, root hairs also play a critical role as the site of infection by symbiotic nitrogen fixing rhizobia, leading to the formation of a novel organ, the nodule. The initial steps in the rhizobia-root hair infection process are known to involve specific receptor kinases and subsequent kinase cascades. Here, we characterize the phosphoproteome of the root hairs and the corresponding stripped roots (i.e., roots from which root hairs were removed) during rhizobial colonization and infection to gain insight into the molecular mechanism of root hair cell biology. We chose soybean (Glycine max L.), one of the most important crop plants in the legume family, for this study because of its larger root size, which permits isolation of sufficient root hair material for phosphoproteomic analysis. Phosphopeptides derived from root hairs and stripped roots, mock inoculated or inoculated with the soybean-specific rhizobium Bradyrhizobium japonicum, were labeled with the isobaric tag 8-plex ITRAQ, enriched using Ni-NTA magnetic beads and subjected to nRPLC-MS/MS analysis using HCD and decision tree guided CID/ETD strategy. A total of 1,625 unique phosphopeptides, spanning 1,659 non-redundant phosphorylation sites, were detected from 1,126 soybean phosphoproteins. Among them, 273 phosphopeptides corresponding to 240 phosphoproteins were found to be significantly regulated (>1.5 fold abundance change) in response to inoculation with B. japonicum. The data reveal unique features of the soybean root hair phosphoproteome, including root hair and stripped root-specific phosphorylation suggesting a complex network of kinase-substrate and phosphatase-substrate interactions in response to rhizobial inoculation.

  1. Quantitative Phosphoproteomic Analysis of Soybean Root Hairs Inoculated with Bradyrhizobium japonicum*

    Science.gov (United States)

    Nguyen, Tran Hong Nha; Brechenmacher, Laurent; Aldrich, Joshua T.; Clauss, Therese R.; Gritsenko, Marina A.; Hixson, Kim K.; Libault, Marc; Tanaka, Kiwamu; Yang, Feng; Yao, Qiuming; Paša-Tolić, Ljiljana; Xu, Dong; Nguyen, Henry T.; Stacey, Gary

    2012-01-01

    Root hairs are single hair-forming cells on roots that function to increase root surface area, enhancing water and nutrient uptake. In leguminous plants, root hairs also play a critical role as the site of infection by symbiotic nitrogen fixing rhizobia, leading to the formation of a novel organ, the nodule. The initial steps in the rhizobia-root hair infection process are known to involve specific receptor kinases and subsequent kinase cascades. Here, we characterize the phosphoproteome of the root hairs and the corresponding stripped roots (i.e. roots from which root hairs were removed) during rhizobial colonization and infection to gain insight into the molecular mechanism of root hair cell biology. We chose soybean (Glycine max L.), one of the most important crop plants in the legume family, for this study because of its larger root size, which permits isolation of sufficient root hair material for phosphoproteomic analysis. Phosphopeptides derived from root hairs and stripped roots, mock inoculated or inoculated with the soybean-specific rhizobium Bradyrhizobium japonicum, were labeled with the isobaric tag eight-plex iTRAQ, enriched using Ni-NTA magnetic beads and subjected to nanoRPLC-MS/MS1 analysis using HCD and decision tree guided CID/ETD strategy. A total of 1625 unique phosphopeptides, spanning 1659 nonredundant phosphorylation sites, were detected from 1126 soybean phosphoproteins. Among them, 273 phosphopeptides corresponding to 240 phosphoproteins were found to be significantly regulated (>1.5-fold abundance change) in response to inoculation with B. japonicum. The data reveal unique features of the soybean root hair phosphoproteome, including root hair and stripped root-specific phosphorylation suggesting a complex network of kinase-substrate and phosphatase-substrate interactions in response to rhizobial inoculation. PMID:22843990

  2. Quantitative phosphoproteomic analysis of soybean root hairs inoculated with Bradyrhizobium japonicum.

    Science.gov (United States)

    Nguyen, Tran Hong Nha; Brechenmacher, Laurent; Aldrich, Joshua T; Clauss, Therese R; Gritsenko, Marina A; Hixson, Kim K; Libault, Marc; Tanaka, Kiwamu; Yang, Feng; Yao, Qiuming; Pasa-Tolić, Ljiljana; Xu, Dong; Nguyen, Henry T; Stacey, Gary

    2012-11-01

    Root hairs are single hair-forming cells on roots that function to increase root surface area, enhancing water and nutrient uptake. In leguminous plants, root hairs also play a critical role as the site of infection by symbiotic nitrogen fixing rhizobia, leading to the formation of a novel organ, the nodule. The initial steps in the rhizobia-root hair infection process are known to involve specific receptor kinases and subsequent kinase cascades. Here, we characterize the phosphoproteome of the root hairs and the corresponding stripped roots (i.e. roots from which root hairs were removed) during rhizobial colonization and infection to gain insight into the molecular mechanism of root hair cell biology. We chose soybean (Glycine max L.), one of the most important crop plants in the legume family, for this study because of its larger root size, which permits isolation of sufficient root hair material for phosphoproteomic analysis. Phosphopeptides derived from root hairs and stripped roots, mock inoculated or inoculated with the soybean-specific rhizobium Bradyrhizobium japonicum, were labeled with the isobaric tag eight-plex iTRAQ, enriched using Ni-NTA magnetic beads and subjected to nanoRPLC-MS/MS1 analysis using HCD and decision tree guided CID/ETD strategy. A total of 1625 unique phosphopeptides, spanning 1659 nonredundant phosphorylation sites, were detected from 1126 soybean phosphoproteins. Among them, 273 phosphopeptides corresponding to 240 phosphoproteins were found to be significantly regulated (>1.5-fold abundance change) in response to inoculation with B. japonicum. The data reveal unique features of the soybean root hair phosphoproteome, including root hair and stripped root-specific phosphorylation suggesting a complex network of kinase-substrate and phosphatase-substrate interactions in response to rhizobial inoculation.

  3. Neospora caninum tachyzoites inoculated by the conjunctival route are not vertically transmitted in pregnant cattle: a descriptive study.

    Science.gov (United States)

    Moore, D P; Alvarez-García, G; Chiapparrone, M L; Regidor-Cerrillo, J; Lischinsky, L H; de Yaniz, M G; Odeón, A C; Ortega-Mora, L M; Campero, C M

    2014-01-17

    The aim of this study was to evaluate whether Neospora caninum tachyzoites (Nc-1) inoculated by the conjunctival route in pregnant cows were able to generate infection in their fetuses. Group 1 contained 2 naturally infected cows; group 2 contained two cows inoculated intravenously with 2.5 × 10(8) tachyzoites, group 3 contained two cows inoculated with 2.5 × 10(8) tachyzoites by the conjunctival route, and group 4 contained two uninfected control cows. The four inoculated cows from groups 2 and 3 were challenged at 23 weeks of gestation. An indirect fluorescent antibody test (IFAT), recombinant NcGRA7-based ELISA, ELISA for IgG subisotypes and Western blot analysis were assessed to characterize the humoral immune response in dams. Sera from their fetuses were tested also using Western blot analysis. Routine microscopic evaluation of H&E stained fetal tissues was made and any fetal tissues and placentas with lesions compatible with Neospora-infection were processed by immunohistochemistry (IHC). DNA extraction from fresh and formalin-fixed, paraffin-embedded fetal tissues were tested by nested PCR. All dams from groups 1, 2 and 3 were seropositive by IFAT, rNcGRA7-based-ELISA and Western blot. IgG1/IgG2 ratios were ≤ 1 at weeks 27 and 29 of gestation. Only fetuses from groups 1 and 2 developed N. caninum specific antibodies by Western blot. Histopathological lesions compatible with those caused by N. caninum were observed in fetuses from groups 1 and 2. N. caninum cysts and tachyzoites were observed by IHC on fetal tissues from groups 1 and 2. Only fetal samples from group 2 were positive by PCR. Further work is needed not only to characterize the cellular immune response but also to clarify the consequences on the dam after conjunctival inoculation of N. caninum tachyzoites. This study shows that N. caninum tachyzoites inoculated by the conjunctival route were not vertically transmitted in pregnant cows.

  4. Inducing Resistance to Conspiracy Theory Propaganda: Testing Inoculation and Metainoculation Strategies

    Science.gov (United States)

    Banas, John A.; Miller, Gregory

    2013-01-01

    This investigation examined the boundaries of inoculation theory by examining how inoculation can be applied to conspiracy theory propaganda as well as inoculation itself (called metainoculation). A 3-phase experiment with 312 participants compared 3 main groups: no-treatment control, inoculation, and metainoculation. Research questions explored…

  5. Inoculation methods of Colletotrichum lindemuthianum in seeds of Phaseolus vulgaris

    OpenAIRE

    Patricia Migliorini; Keilor da Rosa Dorneles; Gustavo Fonseca Rodrigues; Gabriele de Paula; Lilian Vanussa Madruga de Tunes

    2017-01-01

    http://dx.doi.org/10.5007/2175-7925.2017v30n1p37 Anthracnose caused by Colletotrichum lindemuthianum (Sacc. & Magnus) Scrib. is one of the most serious diseases of bean crops. Inoculation techniques are important to evaluate the resistance of genotypes and control methods at early stages of seedling development. Due to the need for infected seedlings, the objective of the present study was to test different inoculation methods of C. lindemuthianum on bean seeds to verify the the impact on...

  6. In vitro study of 3D PLGA/n-HAp/β-TCP composite scaffolds with etched oxygen plasma surface modification in bone tissue engineering

    Science.gov (United States)

    Roh, Hee-Sang; Jung, Sang-Chul; Kook, Min-Suk; Kim, Byung-Hoon

    2016-12-01

    Three-dimensional (3D) scaffolds have many advantageous properties for bone tissue engineering application, due to its controllable properties such as pore size, structural shape and interconnectivity. In this study, effects on oxygen plasma surface modification and adding of nano-hydroxyapatite (n-HAp) and β-tricalcium phosphate (β-TCP) on the 3D PLGA/n-HAp/β-TCP scaffolds for improving preosteoblast cell (MC3T3-E1) adhesion, proliferation and differentiation were investigated. The 3D PLGA/n-HAp/β-TCP scaffolds were fabricated by 3D Bio-Extruder equipment. The 3D scaffolds were prepared with 0°/90° architecture and pore size of approximately 300 μm. In addition 3D scaffolds surface were etched by oxygen plasma to enhance the hydrophilic property and surface roughness. After oxygen plasma treatment, the surface chemistry and morphology were investigated by Fourier transform infrared spectroscopy, scanning electron microscopy, and atomic force microscopy. And also hydrophilic property was measured by contact angle. The MC3T3-E1 cell proliferation and differentiation were investigated by MTT assay and ALP activity. In present work, the 3D PLGA/HAp/beta-TCP composite scaffold with suitable structure for the growth of osteoblast cells was successfully fabricated by 3D rapid prototyping technique. The surface hydrophilicity and roughness of 3D scaffold increased by oxygen plasma treatment had a positive effect on cell adhesion, proliferation, and differentiation. Furthermore, the differentiation of MC3T3-E1 cell was significantly enhanced by adding of n-HAp and β-TCP on 3D PLGA scaffold. As a result, combination of bioceramics and oxygen plasma treatment showed a synergistic effect on biocompatibility of 3D scaffolds. This result confirms that this technique was useful tool for improving the biocompatibility in bone tissue engineering application.

  7. Effect of actinobacteria agent inoculation methods on cellulose degradation during composting based on redundancy analysis.

    Science.gov (United States)

    Zhao, Yue; Lu, Qian; Wei, Yuquan; Cui, Hongyang; Zhang, Xu; Wang, Xueqin; Shan, Si; Wei, Zimin

    2016-11-01

    In this study, actinobacteria agent including Streptomyces sp. and Micromonospora sp. were inoculated during chicken manure composting by different inoculation methods. The effect of different treatments on cellulose degradation and the relationship between inoculants and indigenous actinobacteria were investigated during composting. The results showed that inoculation in different stages of composting all improved the actinobacteria community diversity particularly in the cooling stage of composting (M3). Moreover, inoculation could distinctly accelerate the degradation of organic matters (OM) especially celluloses. Redundancy analysis indicated that the correlation between indigenous actinobacteria and degradation of OM and cellulose were regulated by inoculants and there were significant differences between different inoculation methods. Furthermore, synergy between indigenous actinobacteria and inoculants for degradation of OM and cellulose in M3 was better than other treatments. Conclusively, we suggested an inoculation method to regulate the indigenous actinobacteria based on the relationship between inoculants and indigenous actinobacteria and degradation content.

  8. Susceptibility of alpine ibex to conjunctivitis caused by inoculation of a sheep-strain of Mycoplasma conjunctivae.

    Science.gov (United States)

    Giacometti, M; Nicolet, J; Frey, J; Krawinkler, M; Meier, W; Welle, M; Johansson, K E; Degiorgis, M P

    1998-04-15

    We evaluated the susceptibility of alpine ibex (Capra ibex ibex) to mycoplasmal conjunctivitis induced by a strain of Mycoplasma conjunctivae isolated from domestic sheep by inoculation of three alpine ibexes with 1.2 x 10(6) colony forming units of M. conjunctivae in the conjunctival sac of both eyes. One more ibex was exposed to the infection by contact. Experimental animals were free of M. conjunctivae and ocular Chlamydia infection before inoculation. Conjunctivitis and serous to mucous lachrymation became apparent in all four ibexes. Clinical signs began within 2 days in inoculated animals and 22 days after the beginning of the experiment in the contact ibex. M. conjunctivae was demonstrated up to the 63th day post-inoculation by cultural and PCR-methods. After 63 days, histopathologic examination revealed nearly normal ocular tissues, and M. conjunctivae could be detected from two eyes only. No other infectious agents which might cause conjunctivitis or keratitis, including Chlamydia psittaci and Branhamella ovis, were involved. Our investigation indicates that sheep-strains of M. conjunctivae can induce conjunctivitis in alpine ibex, thus showing pathogenicity of this organism for Caprinae species other than domestic sheep and goats.

  9. Immuno disc assay for screening duck hepatitis B surface antigen in serum, liver tissue and cultured hepatocytes

    NARCIS (Netherlands)

    G.A. de Wilde (G.); R.A. Heijtink

    1993-01-01

    textabstractAn immuno disc assay (IDA) for semi-quantitative analysis of the surface antigen (DHBsAg) of duck hepatitis B virus (DHBV) is described. Unpurified antigen preparations were adsorbed onto punched-out nitrocellulose membrane discs. Rabbit antiserum raised against serum-derived

  10. Physiological, structural and molecular traits activated in strawberry plants after inoculation with the plant growth-promoting bacterium Azospirillum brasilense REC3.

    Science.gov (United States)

    Guerrero-Molina, M F; Lovaisa, N C; Salazar, S M; Martínez-Zamora, M G; Díaz-Ricci, J C; Pedraza, R O

    2015-05-01

    The plant growth-promoting strain REC3 of Azospirillum brasilense, isolated from strawberry roots, prompts growth promotion and systemic protection against anthracnose disease in this crop. Hence, we hypothesised that A. brasilense REC3 can induce different physiological, structural and molecular responses in strawberry plants. Therefore, the aim of this work was to study these traits activated in Azospirillum-colonised strawberry plants, which have not been assessed until now. Healthy, in vitro micropropagated plants were root-inoculated with REC3 under hydroponic conditions; root and leaf tissues were sampled at different times, and oxidative burst, phenolic compound content, malondialdehyde (MDA) concentration, callose deposition, cell wall fortification and gene expression were evaluated. Azospirillum inoculation enhanced levels of soluble phenolic compounds after 12 h post-inoculation (hpi), while amounts of cell wall bound phenolics were similar in inoculated and control plants. Other early responses activated by REC3 (at 24 hpi) were a decline of lipid peroxidation and up-regulation of strawberry genes involved in defence (FaPR1), bacterial recognition (FaFLS2) and H₂O₂ depuration (FaCAT and FaAPXc). The last may explain the apparent absence of oxidative burst in leaves after bacterial inoculation. Also, REC3 inoculation induced delayed structural responses such as callose deposition and cell wall fortification (at 72 hpi). Results showed that A. brasilense REC3 is capable of exerting beneficial effects on strawberry plants, reinforcing their physiological and cellular characteristics, which in turns contribute to improve plant performance.

  11. Development of Polioencephalomyelitis in Cesarean-Derived Colostrum-Deprived Pigs Following Experimental Inoculation with Either Teschovirus A Serotype 2 or Serotype 11.

    Science.gov (United States)

    Matias Ferreyra, Franco; Arruda, Bailey; Stevenson, Gregory; Schwartz, Kent; Madson, Darin; Yoon, Kyoung-Jin; Zhang, Jianqiang; Piñeyro, Pablo; Chen, Qi; Arruda, Paulo

    2017-07-08

    Teschovirus encephalomyelitis is a sporadic disease associated with Teschovirus A (PTV) serotype 1 and, less frequently, other serotypes. In recent years, the number of cases submitted to the Iowa State University Veterinary Diagnostic Laboratory with a history of posterior paresis has increased. Submission histories from various regions of the United States suggest a trend for clinical disease to persist in herds and affect a wider age-range of pigs than historically reported. Polioencephalitis and/or myelitis was consistently present and PTV was detected in affected neural tissue by PCR in a portion of cases. Sequencing from two clinical cases identified PTV-2 and PTV-11. To assess neuropathogenicity of these isolates, 5-week-old cesarean derived and colostrum-deprived pigs were assigned to three groups: negative control (n = 4), PTV-2-inoculated (n = 7), and PTV-11-inoculated (n = 7). Three PTV-2-inoculated pigs developed mild incoordination of the hind limbs, one of which progressed to posterior ataxia. While all PTV-11-inoculated pigs showed severe neurological signs consistent with Teschovirus encephalomyelitis, no evidences of neurological signs were observed in sham-inoculated animals. All PTV-2- and PTV-11-inoculated pigs had microscopic lesions consistent with Teschovirus encephalomyelitis. To our knowledge, this is the first description of PTV-11 and experimental study demonstrating the neuropathogenicity of PTV-11 in the United States.

  12. Pathogenesis of infection with 2009 pandemic H1N1 influenza virus in isogenic guinea pigs after intranasal or intratracheal inoculation.

    Science.gov (United States)

    Wiersma, Lidewij C M; Vogelzang-van Trierum, Stella E; van Amerongen, Geert; van Run, Peter; Nieuwkoop, Nella J; Ladwig, Mechtild; Banneke, Stefanie; Schaefer, Hubert; Kuiken, Thijs; Fouchier, Ron A M; Osterhaus, Albert D M E; Rimmelzwaan, Guus F

    2015-03-01

    To elucidate the pathogenesis and transmission of influenza virus, the ferret model is typically used. To investigate protective immune responses, the use of inbred mouse strains has proven invaluable. Here, we describe a study with isogenic guinea pigs, which would uniquely combine the advantages of the mouse and ferret models for influenza virus infection. Strain 2 isogenic guinea pigs were inoculated with H1N1pdm09 influenza virus A/Netherlands/602/09 by the intranasal or intratracheal route. Viral replication kinetics were assessed by determining virus titers in nasal swabs and respiratory tissues, which were also used to assess histopathologic changes and the number of infected cells. In all guinea pigs, virus titers peaked in nasal secretions at day 2 after inoculation. Intranasal inoculation resulted in higher virus excretion via the nose and higher virus titers in the nasal turbinates than intratracheal inoculation. After intranasal inoculation, infectious virus was recovered only from nasal epithelium; after intratracheal inoculation, it was recovered also from trachea, lung, and cerebrum. Histopathologic changes corresponded with virus antigen distribution, being largely limited to nasal epithelium for intranasally infected guinea pigs and more widespread in the respiratory tract for intratracheally infected guinea pigs. In summary, isogenic guinea pigs show promise as a model to investigate the role of humoral and cell-mediated immunities to influenza and their effect on virus transmission.

  13. Mycorrhizal inoculation of pecan seedlings with some marketable truffles

    Directory of Open Access Journals (Sweden)

    Gian M. Benucci

    2013-12-01

    Full Text Available Pecan is the common name of Carya illinoinensis (Wangenh. K. Koch, an ectomycorrhizal tree native to North America, also frequently known as hickory. Mycorrhizal inoculations of pecan seedlings with: Tuber aestivum Vittad., T. borchii Vittad., T. indicum Cooke & Massee, and T. lyonii Butters are described and discussed.

  14. Erythema at BCG Inoculation Site in Kawasaki Disease Patients.

    Science.gov (United States)

    Rezai, Mohammad Sadegh; Shahmohammadi, Soheila

    2014-08-01

    Kawasaki disease is an acute, self-limiting childhood systemic vasculitis with unknown etiology. Because there is no diagnostic test for Kawasaki disease, the diagnosis is based on clinical criteria. An important clinical sign that is not included in the classical clinical criteria for Kawasaki disease is a reaction at the Bacille Calmette-Guérin inoculation site that are present in about 30-50% of Kawasaki disease patients. of this review was to highlight the usefulness of the inflammation at the Bacille Calmette-Guérin inoculation site for early diagnosis of Kawasaki disease, we conducted a literature review on Medline in PubMed area, Google scholar, Magiran and Scientific Information Database using the search terms "Kawasaki disease, Erythema, BCG, inoculation site, children, cardiac complications, coronary artery lesion, aneurysm, incomplete Kawasaki in 2013. A total of 15 articles had been found. Erythema at the Bacille Calmette-Guérin inoculation site was found in 49.87% of Kawasaki disease patients. Coronary artery abnormalities were found in 10.3% of cases. According to this review, BCGitis is more prevalent than cervical lymphadenopathy and rash and it can be a useful criterion in the diagnosis of incomplete Kawasaki disease in cases not fulfills the classic criteria of at least four of the five findings.

  15. Effects of bacterial inoculation on the fermentation characteristics ...

    African Journals Online (AJOL)

    ... and CO2, pH, hours for temperature to rise above 2 ºC, yeasts and moulds were ... Further work is needed to determine the effects of soybean silage on growth ... Keywords: Cultivars, forages, inoculants, legumes, nutrients, silage, soybean ...

  16. The effect of rhizobacterial inoculation on growth and nutrient ...

    African Journals Online (AJOL)

    PRECIOUS

    2009-11-02

    Nov 2, 2009 ... 2Crop Science, Faculty of Agriculture, Universiti Putra Malysia, ... was conducted to observe the effect of plant growth promoting ... Key words: Banana, rhizobacteria, root stimulation, growth, nutrient uptake. ... Biofertilizer, inoculants of microorganisms, an alter- ... ensure a sustainable banana production.

  17. Inoculating against Pro-Plagiarism Justifications: Rational and Affective Strategies

    Science.gov (United States)

    Compton, Josh; Pfau, Michael

    2008-01-01

    Student plagiarism continues to threaten academic integrity. This investigation assessed whether an inoculation message strategy could combat university plagiarism by protecting student attitudes against pro-plagiarism justification arguments. Additionally, we sought theoretical confirmation of previous findings on involvement and accessibility in…

  18. New Inoculants Containing Lactic Bacteria Applied in Forage Ensiling

    Directory of Open Access Journals (Sweden)

    Teodor Vintila

    2010-05-01

    Full Text Available In a first study, the capacity of lactic bacteria to accumulate biomass in different culture media and temperatures was tested and the biosynthesis parameters were established. In the second study, the strains producing the highest quantity of biomass and determining the most rapid pH drop in culture medium were conditioned in solid supports. The obtained solid products containing lactic bacteria were used to inoculate different types of forages. Ensilage was carried out in laboratory silos made from O2-impermeable plastic flasks, vacuumed using a vacuum pump. The experiment was 2 x 2 factorial with two types of forage (alfalfa and sorghum, each of them inoculated and not inoculated with lactic bacteria. The evolution of lactic bacteria, pH value, and the concentration in volatile acids was verified. In the third experiment, lactic bacteria were used to inoculate silages in farm conditions. The obtained results recommend the tested strain for the improvement of preserving conditions and nutritive value in ensiled forages.

  19. Inoculating against Pro-Plagiarism Justifications: Rational and Affective Strategies

    Science.gov (United States)

    Compton, Josh; Pfau, Michael

    2008-01-01

    Student plagiarism continues to threaten academic integrity. This investigation assessed whether an inoculation message strategy could combat university plagiarism by protecting student attitudes against pro-plagiarism justification arguments. Additionally, we sought theoretical confirmation of previous findings on involvement and accessibility in…

  20. Inoculation methods of Colletotrichum lindemuthianum in seeds of Phaseolus vulgaris

    Directory of Open Access Journals (Sweden)

    Patricia Migliorini

    2017-02-01

    Full Text Available Anthracnose caused by Colletotrichum lindemuthianum (Sacc. & Magnus Scrib. is one of the most serious diseases of bean crops. Inoculation techniques are important to evaluate the resistance of genotypes and control methods at early stages of seedling development. Due to the need for infected seedlings, the objective of the present study was to test different inoculation methods of C. lindemuthianum on bean seeds to verify the the impact on the emergence and early development of seedlings. Seeds of BRS Expedito were inoculated by spore suspension, contact with the pathogen in a water restricted medium, and with PDA discs containing mycelium and spores of the fungus. The experimental design was completely randomized with eight repetitions. Emergence, shoot length and dry mass, and incidence of symptoms of the disease on the hypocotyl and leaf were evaluated. The inoculation methods were effective at causing the infection of the pathogen in the bean seedlings, without affecting emergence, and contact with the pathogen in a water restricted medium was the most effective method.

  1. Inoculation methods of Colletotrichum lindemuthianum in seeds of Phaseolus vulgaris

    Directory of Open Access Journals (Sweden)

    Patricia Migliorini

    2017-02-01

    Full Text Available http://dx.doi.org/10.5007/2175-7925.2017v30n1p37 Anthracnose caused by Colletotrichum lindemuthianum (Sacc. & Magnus Scrib. is one of the most serious diseases of bean crops. Inoculation techniques are important to evaluate the resistance of genotypes and control methods at early stages of seedling development. Due to the need for infected seedlings, the objective of the present study was to test different inoculation methods of C. lindemuthianum on bean seeds to verify the the impact on the emergence and early development of seedlings. Seeds of BRS Expedito were inoculated by spore suspension, contact with the pathogen in a water restricted medium, and with PDA discs containing mycelium and spores of the fungus. The experimental design was completely randomized with eight repetitions. Emergence, shoot length and dry mass, and incidence of symptoms of the disease on the hypocotyl and leaf were evaluated. The inoculation methods were effective at causing the infection of the pathogen in the bean seedlings, without affecting emergence, and contact with the pathogen in a water restricted medium was the most effective method.

  2. smallholder farmers' use and profitability of legume inoculants in ...

    African Journals Online (AJOL)

    ACSS

    In Kenya, wide-scale adoption of this innovation among the smallholder ... Rhizobia inoculant, a product of Kenya, and its profitability in smallholder farms. Data were collected from ... Rhizobia may pose a barrier to infection and subsequent ... small-scale farmers in western Kenya. .... on soybean production. Gross margin is ...

  3. Preparation of hydrophilic poly(lactic acid) tissue engineering scaffold via (PLA)-(PLA-b-PEG)-(PEG) solution casting and thermal-induced surface structural transformation.

    Science.gov (United States)

    Zhu, Xiaomin; Zhong, Tian; Huang, Ran; Wan, Ajun

    2015-01-01

    Porous poly(lactic acid) (PLA) tissue engineering scaffolds with a hydrophilic surface assembled by polyethylene glycol aggregations were prepared by the solvent casting/particulate leaching method from (PLA)-(PLA-b-PEG)-(PEG) blend solution, where the PLA-b-PEG block polymer serves as an amphiphilic glue between two phases. A thermal recrystallization process was inserted before leaching to induce a phase separation, which subsequently squeezes out PEG to form a hydrophilic shell. Characterizations of XRD and DSC indicated the composition and mixing states of materials. The water contact angle test qualitatively presented the excellent hydrophilicity compared to the pure PLA or PLA-PEG simple blend scaffold. The scanning electron microscope results confirmed the formation of porous structure of [Formula: see text] pore size, with an observable phase separation on the surface. The scaffold was degraded in PBS at [Formula: see text], and the degradation exhibits a three-stage behavior, which evidenced the amphiphilically glued phase separations.

  4. Suppressive properties of composts may be improved by microbial inoculation

    Directory of Open Access Journals (Sweden)

    Polina Galitskaya

    2016-04-01

    Full Text Available The use of suppressive composts instead of traditional fertilizers and pesticides is becoming more commonplace in modern agricultural biotechnology. In this study, we investigated the possibility of obtaining composts with suppressive properties from widespread agricultural wastes produced in tatarstan (russia. Straw and corn wastes, as well as chicken, cattle and swine manure were used to prepare 11 two- and three-component mixtures for further composting. It was found that mesophilic phase of composting lasted for 1 to 3 days for all the waste mixtures; the thermophilic phase lasted from 2 to 35 days, and was characterized by significant fluctuations in temperature, i.e. From 27°с to 59°с. In the initial mixture, the dissolved organic carbon (doc content was between 10 and 62 mg kg-1 ; it fell significantly on day 13, and then continuously decreased up to day 102, and subsequently remained low. For all the mixtures, maximal respiration activity was observed in the beginning of composting (231.9 mg со2-сg -1 . After 23 days, this parameter decreased significantly, and fluctuations subsided. The phytotoxicity of the initial compost mixtures varied from 18% (sw+sm to 100% (cw+chm+sm, cw+chm; however, the trends in the dynamics were similar. After 120 days of composting, 5 of 11 samples were not phytotoxic, and the others were slightly toxic. After 120 days of composting, each mixture was divided into two parts; one was inoculated with a biopreparation consisting of four microbial strains (trichoderma asperellum t203, pseudomonas putida pcl1760, pseudomonas fluorescens wcs365 and streptomyces spp., and the other part was not inoculated. Both parts were composted under equivalent conditions for 60 days. Inoculation led to a slightly shorter period of increasing doc and respiration activity. However, after 3-10 days, no differences were observed between the mixtures. Inoculation did not influence the temperature profile or phytotoxicity of

  5. Surface survival and internalization of salmonella through natural cracks on developing cantaloupe fruits, alone or in the presence of the melon wilt pathogen Erwinia tracheiphila.

    Directory of Open Access Journals (Sweden)

    Dhiraj Gautam

    Full Text Available Outbreaks of foodborne illness attributed to the consumption of Salmonella-tainted cantaloupe have occurred repeatedly, but understanding of the ecology of Salmonella on cantaloupe fruit surfaces is limited. We investigated the interactions between Salmonella enterica Poona, the plant pathogenic bacterium Erwinia tracheiphila, and cantaloupe fruit. Fruit surfaces were inoculated at the natural cracking stage by spreading S. enterica and E. tracheiphila, 20 µl at 107 cfu/ml, independently or together, over a 2×2 cm rind area containing a crack. Microbial and microscopic analyses were performed at 0, 9 and 24 days post inoculation (DPI. Even at 24 DPI (fruit maturity S. enterica was detected on 14% and 40% of the fruit inoculated with S. enterica alone and the two-pathogen mixture, respectively. However, the population of S. enterica declined gradually after initial inoculation. E. tracheiphila, inoculated alone or together with Salmonella, caused watersoaked lesions on cantaloupe fruit; but we could not conclude in this study that S. enterica survival on the fruit surface was enhanced by the presence of those lesions. Of fruit inoculated with E. tracheiphila alone and sampled at 24 DPI, 61% had watersoaked lesions on the surface. In nearly half of those symptomatic fruits the watersoaking extended into the sub-rind mesocarp, and E. tracheiphila was recovered from that tissue in 50% of the symptomatic fruit. In this work, E. tracheiphila internalized through natural cracks on developing fruits. S. enterica was never detected in the fruit interior (ca. 2-3 mm below rind surface under the limited conditions of our experiments, but the possibility that it, or other human pathogens that contaminate fresh produce, might also do so should be investigated under a wider range of conditions and produce types.

  6. Surface survival and internalization of salmonella through natural cracks on developing cantaloupe fruits, alone or in the presence of the melon wilt pathogen Erwinia tracheiphila.

    Science.gov (United States)

    Gautam, Dhiraj; Dobhal, Shefali; Payton, Mark E; Fletcher, Jacqueline; Ma, Li Maria

    2014-01-01

    Outbreaks of foodborne illness attributed to the consumption of Salmonella-tainted cantaloupe have occurred repeatedly, but understanding of the ecology of Salmonella on cantaloupe fruit surfaces is limited. We investigated the interactions between Salmonella enterica Poona, the plant pathogenic bacterium Erwinia tracheiphila, and cantaloupe fruit. Fruit surfaces were inoculated at the natural cracking stage by spreading S. enterica and E. tracheiphila, 20 µl at 107 cfu/ml, independently or together, over a 2×2 cm rind area containing a crack. Microbial and microscopic analyses were performed at 0, 9 and 24 days post inoculation (DPI). Even at 24 DPI (fruit maturity) S. enterica was detected on 14% and 40% of the fruit inoculated with S. enterica alone and the two-pathogen mixture, respectively. However, the population of S. enterica declined gradually after initial inoculation. E. tracheiphila, inoculated alone or together with Salmonella, caused watersoaked lesions on cantaloupe fruit; but we could not conclude in this study that S. enterica survival on the fruit surface was enhanced by the presence of those lesions. Of fruit inoculated with E. tracheiphila alone and sampled at 24 DPI, 61% had watersoaked lesions on the surface. In nearly half of those symptomatic fruits the watersoaking extended into the sub-rind mesocarp, and E. tracheiphila was recovered from that tissue in 50% of the symptomatic fruit. In this work, E. tracheiphila internalized through natural cracks on developing fruits. S. enterica was never detected in the fruit interior (ca. 2-3 mm below rind surface) under the limited conditions of our experiments, but the possibility that it, or other human pathogens that contaminate fresh produce, might also do so should be investigated under a wider range of conditions and produce types.

  7. Collagen hydrogels incorporated with surface-aminated mesoporous nanobioactive glass: Improvement of physicochemical stability and mechanical properties is effective for hard tissue engineering.

    Science.gov (United States)

    El-Fiqi, Ahmed; Lee, Jae Ho; Lee, Eun-Jung; Kim, Hae-Won

    2013-12-01

    Collagen (Col) hydrogels have poor physicochemical and mechanical properties and are susceptible to substantial shrinkage during cell culture, which limits their potential applications in hard tissue engineering. Here, we developed novel nanocomposite hydrogels made of collagen and mesoporous bioactive glass nanoparticles (mBGns) with surface amination, and addressed the effects of mBGn addition (Col:mBG = 2:1, 1:1 and 1:2) and its surface amination on the physicochemical and mechanical properties of the hydrogels. The amination of mBGn was shown to enable chemical bonding with collagen molecules. As a result, the nanocomposite hydrogels exhibited a significantly improved physicochemical and mechanical stability. The hydrolytic and enzymatic degradation of the Col-mBGn hydrogels were slowed down due to the incorporation of mBGn and its surface amination. The mechanical properties of the hydrogels, specifically the resistance to loading as well as the stiffness, significantly increased with the addition of mBGn and its aminated form, as assessed by a dynamic mechanical analysis. Mesenchymal stem cells cultivated within the Col-mBGn hydrogels were highly viable, with enhanced cytoskeletal extensions, due to the addition of surface aminated mBGn. While the Col hydrogel showed extensive shrinkage (down to ∼20% of initial size) during a few days of culture, the shrinkage of the mBGn-added hydrogel was substantially reduced, and the aminated mBGn-added hydrogel had no observable shrinkage over 21 days. Results demonstrated the effective roles of aminated mBGn in significantly improving the physicochemical and mechanical properties of Col hydrogel, which are ultimately favorable for applications in stem cell culture for bone tissue engineering.

  8. Long-term humoral antibody responses by various serologic tests in pigs orally inoculated with oocysts of four strains of Toxoplasma gondii

    DEFF Research Database (Denmark)

    Dubey, J.P.; Andrews, C.D.; Thulliez, P.;

    1997-01-01

    Antibody titers to Toxoplasma gondii were determined in 16 pigs orally inoculated with 1000 or 10000 oocysts of one of the four strains (GT-1, ME-49, TS-2, TC-2) of T. gondii. Pigs were euthanized on postinoculation days 103-875 and their tissues were bioassayed for T. gondii. Antibody titers wer...

  9. The effect of surface modification of a porous TiO2/perlite composite on the ingrowth of bone tissue in vivo.

    Science.gov (United States)

    Erli, Hans-Josef; Rüger, Matthias; Ragoss, Christian; Jahnen-Dechent, Willi; Hollander, Dirk A; Paar, Othmar; von Walter, Matthias

    2006-03-01

    The porous TiO2/perlite composite Ecopore is a synthetic biomaterial with possible clinical application in bone substitution. In our previous work, we demonstrated that surface modification of Ecopore with fibronectin (FN) enhanced spreading and growth of human osteoblasts in vitro. In the present study, we implanted untreated, alkaline-etched and FN-coated Ecopore cylinders into critical size defects of rabbit femora and applied pulsed polychrome sequence staining. After 6 weeks, sections of the implants were investigated via conventional and fluorescence microscopy. A partial ingrowth of bone matrix into the pore system of the Ecopore implants was observed. At the contact zones, the bone appeared to be directly connected to the implant without detectable gaps. Defect healing was complete within 6 weeks, while fibrous tissue generation or inflammation were absent in the implant modification groups, demonstrating basic Ecopore biocompatibility. The mean bone apposition rates within the implant cross-section were 4.1+/-0.6 microm/day (p0.05) in the NaOH-etched group and 10.2+/-5.5% in the unmodified group. In summary, we conclude that bone tissue tolerates Ecopore well and that tissue ingrowth can be enhanced by etching and coating with FN.

  10. A simple, non-invasive and inexpensive method for evaluating the displacement of local tissue surfaces: from vascular changes to muscle contraction.

    Science.gov (United States)

    Roatta, S; Deriu, F; Artusio, E; Passatore, M

    1996-01-01

    This paper describes a non-invasive, very inexpensive method of estimating tissue displacements of various origin that is easy and fast to set up. This technique utilizes an inductive proximity sensor (IPS), which is a non-contact length transducer measuring the distance between its probe and a metal target. Its working principle is based on the electromagnetic coupling originating between the sensor probe, a source of high-frequency magnetic field, and the metal target where parasitic currents take place. The linear working range of the IPS model used here is 0.1 to 6 mm probe-target distance, its resolution is about 2 microns. The IPS has been employed on rabbits and humans to measure the displacement of a target glued to the skin of various body areas with respect to the fixed probe of the sensor. Its high resolution, together with an extensive working range, allows the evaluation of numerous physiological events which produce displacements ranging from 2 microns -- to 9 mm, reflecting either tissue volume changes or movements. In particular, an interesting application is to monitor, through volume variations, the extent and the time course of local vascular modifications induced by manoeuvres which elicit changes in vasomotor tone; vascular filling, tissue swelling etc. Therefore, this measure may be considered a 'surface plethysmography' record. In addition, the contractions of skeletal muscles, under either isotonic or isometric conditions, can be estimated through this sensor. This system may therefore find applications for research purposes and practical demonstrations to students.

  11. Efect of intercellular extracts from banana inoculated leaves with Mycosphaerella fijiensis Morelet, on chloroplast electronic transport of Grande naine (AAA cv.

    Directory of Open Access Journals (Sweden)

    Michel Leiva-Mora

    2003-01-01

    Full Text Available Some foliar pathogens colonize intercellular spaces of damage tissues during infection process, mediated by toxins production and diffusion to kill adjacent healthy cells. Due to the absence of reliable bioassays, the physiologic effects of several phytotoxins are still ignored on cellular membranous systems of the affected cells. In the present work it was extracted the intercellular content from not inoculated and inoculated banana leaves with different Mycosphaerella fijiensis strains. Their effects on chloroplasts of Grande naine cv were evaluated by the absorbance evolution (595 nm of Hill reactive (DCPIP, mixture with 810 ì l of chloroplasts suspension and 99 ì l of the intercellular contents. The electronic exchange on chloroplasts suspension was inhibited by intercellular contents of inoculated leaves. The intercellular contents from leaves inoculated with I1 (high virulence strain had a major inhibiter effect respect to leaves inoculates with G1 strain (low virulence, showing a correspondence between the inhibiter effect of intercellular contents and the affection levels of affected tissues. The procedures used in this work will let to make studies concerned with Mycosphaerella fijiensis-Musa spp interactions and the future breeding programs. Key words: banana breeding, black Sigatoka, host pathogen interaction, physiological bioassays

  12. Influence of Pleurotus ostreatus inoculation on wood degradation and fungal colonization.

    Science.gov (United States)

    Piškur, Barbara; Bajc, Marko; Robek, Robert; Humar, Miha; Sinjur, Iztok; Kadunc, Aleš; Oven, Primož; Rep, Gregor; Al Sayegh Petkovšek, Samar; Kraigher, Hojka; Jurc, Dušan; Pohleven, Franc

    2011-11-01

    The influence of Pleurotus ostreatus inoculation on wood degradation and on fungal community structure was studied. The experiments were performed on an organically poor fly ash deposit covered with a 10 cm layer of beech wood chips inoculated with P. ostreatus isolate ZIM76. Compared to non-inoculated wood chips, inoculation increased the temperatures and relative humidities and, in the first 6 months, accelerated Klason lignin degradation by 9% and also, after 17 months, increased iron translocation into wood chips by 30%. After 6 months, PCR-DGGE showed 22-28 and 13-21 fungal taxa in non-inoculated and P. ostreatus-inoculated beech chips, respectively. The differences in number of taxa and in the fungal community structure (based on Dice coefficient) between non-inoculated and inoculated wood chips diminished with time. The results indicate that the naturally occurring processes of wood degradation are as efficient as those occurring in sites inoculated with P. ostreatus.

  13. Poliomyelitis in transgenic mice expressing CD155 under the control of the Tage4 promoter after oral and parenteral poliovirus inoculation.

    Science.gov (United States)

    Khan, Shaukat; Toyoda, Hidemi; Linehan, Melissa; Iwasaki, Akiko; Nomoto, Akio; Bernhardt, Günter; Cello, Jeronimo; Wimmer, Eckard

    2014-08-01

    An important step in poliovirus (PV) infection by the oral route in humans is replication of the virus in lymphatic tissues of the gastrointestinal (GI) tract, thought to be mainly in the Peyer's patches of the small intestine. No immunocompetent transgenic (tg) mice that express human PV receptor (CD155) under the control of different promoters can be infected orally. The mouse orthologue of human CD155 is Tage4, a protein expressed at the surface of enterocytes and in the Peyer's patches. We describe here the generation of a tg mouse model in which the Tage4 promoter was used to drive expression of the human PV receptor-coding region (Tage4-CD155tg mice). In this model, CD155 expression was observed by immunostaining in different regions in the Peyer's patches but not in their germinal centres. Although a similar pattern of staining was observed between 3- and 6-week-old Tage4-CD155tg mice, poliomyelitis was only seen in the younger mice after PV infection by the oral route. When compared with TgPVR21 mice that expressed CD155 driven by its human promoter, 3-week-old Tage4-CD155tg mice were more susceptible to gut infection and paralysis following feeding with PV. Also, Tage4-CD155tg mice exhibited higher susceptibility to poliomyelitis after parenteral inoculation of PV. Remarkably, the LD50 after intracerebral inoculation of PV was similar in both CD155 tg mouse strains. The CD155 tg mouse model reported here, although moderately susceptible to oral infection, may be suitable to study mechanisms of PV replication in the gastrointestinal tract and to dissect important aspects of PV neuroinvasiveness.

  14. Cast Iron Inoculation Enhanced by Supplementary Oxy-sulfides Forming Elements

    Science.gov (United States)

    Riposan, Iulian; Stan, Stelian; Uta, Valentin; Stefan, Ion

    2017-08-01

    Inoculation is one of the most important metallurgical treatments applied to the molten cast iron immediately prior to casting, to promote solidification without excessive eutectic undercooling, which favors carbides formation usually with undesirable graphite morphologies. The paper focused on the separate addition of an inoculant enhancer alloy [S, O, oxy-sulfides forming elements] with a conventional Ca-FeSi alloy, in the production of gray and ductile cast irons. Carbides formation tendency decreased with improved graphite characteristics as an effect of the [Ca-FeSi + Enhancer] inoculation combination, when compared to other Ca/Ca, Ba/Ca, RE-FeSi alloy treatments. Adding an inoculant enhancer greatly enhances inoculation, lowers inoculant consumption up to 50% or more and avoids the need to use more costly inoculants, such as a rare earth bearing alloy. The Inoculation Specific Factor [ISF] was developed as a means to more realistically measure inoculant treatment efficiency. It compares the ratio between the improved characteristic level and total inoculant consumption for this effect. Addition of any of the commercial inoculants plus the inoculant enhancer offered outstanding inoculation power [increased ISF] even at higher solidification cooling rates, even though the total enhancer addition was at a small fraction of the amount of commercial inoculant used.

  15. Pseudomonas aeruginosa outer membrane vesicles triggered by human mucosal fluid and lysozyme can prime host tissue surfaces for bacterial adhesion

    Directory of Open Access Journals (Sweden)

    Matteo Maria Emiliano Metruccio

    2016-06-01

    Full Text Available Pseudomonas aeruginosa is a leading cause of human morbidity and mortality that often targets epithelial surfaces. Host immunocompromise, or the presence of indwelling medical devices, including contact lenses, can predispose to infection. While medical devices are known to accumulate bacterial biofilms, it is not well understood why resistant epithelial surfaces become susceptible to P. aeruginosa. Many bacteria, including P. aeruginosa, release Outer Membrane Vesicles (OMVs in response to stress that can fuse with host cells to alter their function. Here, we tested the hypothesis that mucosal fluid can trigger OMV release to compromise an epithelial barrier. This was tested using tear fluid and corneal epithelial cells in vitro and in vivo. After 1 h both human tear fluid, and the tear component lysozyme, greatly enhanced OMV release from P. aeruginosa strain PAO1 compared to PBS controls (~100 fold. TEM and SDS-PAGE showed tear fluid and lysozyme-induced OMVs were similar in size and protein composition, but differed from biofilm-harvested OMVs, the latter smaller with fewer proteins. Lysozyme-induced OMVs were cytotoxic to human corneal epithelial cells in vitro and murine corneal epithelium in vivo. OMV exposure in vivo enhanced Ly6G/C expression at the corneal surface, suggesting myeloid cell recruitment, and primed the cornea for bacterial adhesion (~4-fold, P < 0.01. Sonication disrupted OMVs retained cytotoxic activity, but did not promote adhesion, suggesting the latter required OMV-mediated events beyond cell killing. These data suggest that mucosal fluid induced P. aeruginosa OMVs could contribute to loss of epithelial barrier function during medical device-related infections.

  16. Elemental composition of strawberry plants inoculated with the plant growth-promoting bacterium Azospirillum brasilense REC3, assessed with scanning electron microscopy and energy dispersive X-ray analysis.

    Science.gov (United States)

    Guerrero-Molina, M F; Lovaisa, N C; Salazar, S M; Díaz-Ricci, J C; Pedraza, R O

    2014-07-01

    The elemental composition of strawberry plants (Fragaria ananassa cv. Macarena) inoculated with the plant growth-promoting bacterium Azospirillum brasilense REC3, and non-inoculated controls, was studied using scanning electron microscopy (SEM) and energy dispersive X-ray (EDS) analysis. This allowed simultaneous semi-quantification of different elements in a small, solid sample. Plants were inoculated and grown hydroponically in 50% or 100% Hoagland solution, corresponding to limited or optimum nutrient medium, respectively. Bacteria-inoculated plants increased the growth index 45% and 80% compared to controls when grown in 100% and 50% Hoagland solution, respectively. Thus, inoculation with A. brasilense REC3 in a nutrient-limited medium had the strongest effect in terms of increasing both shoot and root biomass and growth index, as already described for Azospirillum inoculated into nutrient-poor soils. SEM-EDS spectra and maps showed the elemental composition and relative distribution of nutrients in strawberry tissues. Leaves contained C, O, N, Na, P, K, Ca and Cu, while roots also had Si and Cl. The organic fraction (C, O and N) accounted for over 96.3% of the total chemical composition; of the mineral fraction, Na had higher accumulation in both leaves and roots. Azospirillum-inoculated and control plants had similar elemental quantities; however, in bacteria-inoculated roots, P was significantly increased (34.33%), which constitutes a major benefit for plant nutrition, while Cu content decreased (35.16%).

  17. Preparation of a non-woven poly(ε-caprolactone) fabric with partially embedded apatite surface for bone tissue engineering applications by partial surface melting of poly(ε-caprolactone) fibers.

    Science.gov (United States)

    Kim, In Ae; Rhee, Sang-Hoon

    2017-03-21

    This article describes a novel method for the preparation of a biodegradable non-woven poly(ε-caprolactone) fabric with a partially embedded apatite surface designed for application as a scaffold material for bone tissue engineering. The non-woven poly(ε-caprolactone) fabric was generated by the electro-spinning technique and then apatite was coated in simulated body fluid after coating the PVA solution containing CaCl2 ·2H2 O. The apatite crystals were partially embedded or fully embedded into the thermoplastic poly(ε-caprolactone) fibers by controlling the degree of poly(ε-caprolactone) fiber surface melting in a convection oven. Identical apatite-coated poly(ε-caprolactone) fabric that did not undergo heat-treatment was used as a control. The features of the embedded apatite crystals were evaluated by FE-SEM, AFM, EDS, and XRD. The adhesion strengths of the coated apatite layers and the tensile strengths of the apatite coated fabrics with and without heat-treatment were assessed by the tape-test and a universal testing machine, respectively. The degree of water absorbance was assessed by adding a DMEM droplet onto the fabrics. Moreover, cell penetrability was assessed by seeding preosteoblastic MC3T3-E1 cells onto the fabrics and observing the degrees of cell penetration after 1 and 4 weeks by staining nuclei with DAPI. The non-woven poly(ε-caprolactone) fabric with a partially embedded apatite surface showed good water absorbance, cell penetrability, higher apatite adhesion strength, and higher tensile strength compared with the control fabric. These results show that the non-woven poly(ε-caprolactone) fabric with a partially embedded apatite surface is a potential candidate scaffold for bone tissue engineering due to its strong apatite adhesion strength and excellent cell penetrability. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A, 2017.

  18. Induction of cytokines in different organs after intranasal inoculation of Campylobacter jejuni in mice

    Directory of Open Access Journals (Sweden)

    Al-Banna Nadia A

    2012-12-01

    Full Text Available Abstract Background Cytokine production and histopathological changes occur in the lungs of mice after intranasal inoculation with Campylobacter jejuni, but the levels of cytokines in different organs to which C. jejuni disseminates have not been studied. Findings Adult BALB/c mice were intranasally inoculated with C. jejuni 81–176 (test or phosphate-buffered saline (control (n=16 per group. The levels of cytokines in the organs (spleen, liver, and small and large intestines to which C. jejuni disseminated were measured by ELISA. Two cytokine patterns were observed. First, increased proinflammatory cytokines, TNF-α, IL-1, and IL-2, were followed by anti-inflammatory cytokines, IL-4 and IL-10 in the spleen and large intestine. Second, in the liver and small intestine, there was a predominant production of anti-inflammatory cytokines, IL-4 and IL-10, with some increase in IL-2 levels. In the spleen and intestines, the levels of pro- and anti-inflammatory cytokines were concurrently increased. Conclusion Dissemination of C. jejuni is associated with the production of different cytokine profiles in different tissues, with the proinflammatory response appearing in the spleen and large intestine at an earlier time point than in the liver and small intestine. The organs produce different cytokine profiles in response to C. jejuni dissemination. These preliminary findings should be confirmed with a study involving a larger group of animals.

  19. Identification of a Divergent Lineage Porcine Pestivirus in Nursing Piglets with Congenital Tremors and Reproduction of Disease following Experimental Inoculation.

    Directory of Open Access Journals (Sweden)

    Bailey L Arruda

    Full Text Available Congenital tremors is a sporadic disease of neonatal pigs characterized by action-related repetitive myoclonus. A majority of outbreaks of congenital tremors have been attributed to an unidentified virus. The objectives of this project were to 1 detect potential pathogen(s in samples from piglets with congenital tremors and 2 develop an infection model to reproduce disease. Using next-generation sequencing, a divergent lineage pestivirus was detected in piglets with congenital tremors. The virus was originally most closely related to a bat pestivirus but is now more closely related to a recently published novel porcine pestivirus provisionally named atypical porcine pestivirus. A quantitative real-time PCR detected the virus in samples from neonatal piglets with congenital tremors from two separate farms, but not in samples from unaffected piglets from the same farm. To fulfill the second objective, pregnant sows were inoculated with either serum containing the pestivirus or PBS (control by intravenous and intranasal routes simultaneously with direct inoculation of fetal amniotic vesicles by ultrasound-guided surgical technique. Inoculations were performed at either 45 or 62 days of gestation. All sows inoculated with the novel pestivirus farrowed piglets affected with congenital tremors while PBS-inoculated control piglets were unaffected. Tremor severity for each piglet was scored from videos taken 0, 1 and 2 days post-farrowing. Tremor severity remained relatively constant from 0 to 2 days post-farrowing for a majority of piglets. The prevalence of congenital tremors in pestivirus-inoculated litters ranged from 57% (4 out of 7 affected piglets to 100% (10 out of 10 affected piglets. The virus was consistently detected by PCR in tissues from piglets with congenital tremors but was not detected in control piglets. Samples positive by PCR in greater than 90% of piglets sampled included brainstem (37 out of 41, mesenteric lymph node (37 out of 41

  20. Comparative analysis of the expression of surface markers on fibroblasts and fibroblast-like cells isolated from different human tissues.

    Science.gov (United States)

    Lupatov, A Yu; Vdovin, A S; Vakhrushev, I V; Poltavtseva, R A; Yarygin, K N

    2015-02-01

    Expression of 20 surface markers was analyzed in cultures of mesenchymal stromal cells of the umbilical cord, fibroblasts from adult and fetal human skin, and fibroblast-like cells of fetal liver was analyzed by fl ow cytometry. The studied cultures did not express hemopoietic cells markers, but were positive for CD73, CD90, and CD105 markers recommended by the International Society of Cell Therapy for the identification of the multipotent mesenchymal stromal cells. Fetal liver fibroblast-like cells were positive for CD54; this marker was absent in skin fibroblast cultures, but was expressed by umbilical cord mesenchymal stromal cells. Further study of these cells revealed a minor subpopulation of cells co-expressing CD24 and CD90 or CD24 and CD54. We hypothesized that these cells probably participate in epithelial mesenchymal transition.

  1. Yeast inoculation as a strategy to improve the physico-chemical and sensory properties of reduced salt fermented sausages produced with entire male fat.

    Science.gov (United States)

    Corral, Sara; Belloch, Carmela; López-Díez, José Javier; Salvador, Ana; Flores, Mónica

    2017-01-01

    Yeast inoculation of dry fermented sausages manufactured with entire male fat was evaluated as a strategy to improve sausage quality. Four different formulations with entire male/gilt back fat and inoculated/non-inoculated with Debaryomyces hansenii were manufactured. The use of entire male back fat produced the highest weight losses, hardness and chewiness in dry sausages. Consumers clearly distinguished samples according to drying time and D. hansenii inoculation while the use of entire/gilt back fat was not highly perceived. The presence of androstenone and skatole was close to their sensory thresholds. Androstenone was not degraded during the process but skatole was affected by yeast inoculation. D. hansenii growth on the surface regulated water release during ripening, reduced hardness and chewiness in entire male sausages and resulted with similar texture to gilt sausages. Yeast inoculation inhibited lipid oxidation providing fruity odours and less oxidized fatty sausages in the sensory analysis. The effectiveness of yeast to mask boar taint was demonstrated by sensory analysis.

  2. Field performance of Alnus cordata Loisel (Italian alder) inoculated with Frankia and VA-mycorrhizal strains in mine-spoil afforestation plots

    Energy Technology Data Exchange (ETDEWEB)

    Lumini, E.; Bosco, M.; Puppi, G.; Isopi, R.; Frattegiani, M.; Burseti, E.; Favili, F.

    1994-05-01

    Mixed stands of {ital Alnus cordata}, {ital Eleagnus} spp. and broad leaf timber trees have been successfully used for reclamation of mine spoils in surface mined areas of central Italy. Trials showed that the use of mycorrhizal planting stock may improve the establishment of plants on mine spoils. Alder seeds were sown in pots filled with peat moss and lignite mine spoils and Fungal strains {ital Glomus mosscae} and {ital G. fasciculatum} were inoculated into the pots. A Frankia strain isolated from {ital A. cordata} was also tested. Plants were then outplanted after measuring shoot height and collar diameter. The planting was done in mid-January among commercial trees ({ital Quercus robur}, {ital Fraxinus oxyphilla}, and {ital Prunus avium}). Plants became highly nodulated in the pots. Mycorrhizal infection was about 30% on inoculate plants but did not occur on uninoculated plants. Aboveground biomass at out-planting was significantly greater in the Frankia inoculated plants. There was generally more N in plant leaves in the inoculate plants than in the uninoculated ones. Plant survival in the field after one winter was 83% on average. One year after planting, the inoculated plants were significantly bigger as were the Frankia inoculated plants. The combination of Frankia and Glomus was very effective (contrary to other studies).

  3. Ceramic filters for bulk inoculation of nickel alloy castings

    Directory of Open Access Journals (Sweden)

    F. Binczyk

    2011-07-01

    Full Text Available The work includes the results of research on production technology of ceramic filters which, besides the traditional filtering function, playalso the role of an inoculant modifying the macrostructure of cast nickel alloys. To play this additional role, filters should demonstratesufficient compression strength and ensure proper flow rate of liquid alloy. The role of an inoculant is played by cobalt aluminateintroduced to the composition of external coating in an amount from 5 to 10 wt.% . The required compression strength (over 1MPa isprovided by the supporting layers, deposited on the preform, which is a polyurethane foam. Based on a two-level fractional experiment24-1, the significance of an impact of various technological parameters (independent variables on selected functional parameters of theready filters was determined. Important effect of the number of the supporting layers and sintering temperature of filters after evaporationof polyurethane foam was stated.

  4. Surface Modification of Biomimetic PLGA-(ASP-PEG) Matrix with RGD-Containing Peptide: a New Non-Viral Vector for Gene Transfer and Tissue Engineering

    Institute of Scientific and Technical Information of China (English)

    GUO Xiaodong; SONG Yulin; ZHENG Qixin; YANG Shuhua; SHAO Zengwu; WU Yongchao; HAO Jie; QUAN Daping

    2006-01-01

    RGD-containing peptide (K16-GRGDSPC), characterized as non-viral gene vectors, was fabricated to modify the surface of PLGA-[ASP-PEG] matrix, which offered the foundation for gene transfer with porous matrix of gene activated later. Peptide was synthesized and matrix was executed into chips A,B and chip C. Chip C was regarded as control. Chips A and B were reacted with cross-linker. Then chip A was reacted with peptide. MS and HPLC were used to detect the MW and purity of peptide. Sulphur, existing on the surface of biomaterials, was detected by XPS. The purity of un-reacted peptide in residual solution was detected by a spectrophotometer. HPLC shows that the peptide purity was 94%-95%, and MS shows that the MW was 2 741.3307. XPS reveals that the binding energy of sulphur was 164 eV and the ratio of carbon to sulphur (C/S) was 99.746∶0.1014 in reacted chip A. The binding energy of sulphur in reacted chip B was 164 eV and 162 eV, C/S was 99.574∶0.4255, and there was no sulphur in chip C. Peptide was manufactured and linked to the surface of biomimetic and 3-D matrix, which offered the possibilities for gene transfer and tissue engineering with this new kind of non-viral gene vector.

  5. Surface functionalization of tissue culture polystyrene plates with hydroxyapatite under body fluid conditions and its effect on differentiation behaviors of mesenchymal stem cells.

    Science.gov (United States)

    Iijima, Kazutoshi; Suzuki, Ryo; Iizuka, Ayako; Ueno-Yokohata, Hitomi; Kiyokawa, Nobutaka; Hashizume, Mineo

    2016-11-01

    The surfaces of polystyrene (PS) cell culture plates were functionalized with hydroxyapatite (HAp) under body fluid conditions utilizing protein adsorption layers and a pretreatment with an alternate soaking process (ASP) using solutions containing calcium and phosphate ions. Adsorption layers of human serum albumin (HSA) formed on the surface of each well of commercial 24-well PS plates by solution processes. CaCl2 and K2HPO4 solutions were alternately added to the wells, the plates were incubated to form the precursors, and this was followed by the addition of simulated body fluid (SBF) and a further incubation for 24h. These treatments resulted in the surfaces of the PS cell culture plates being completely covered with bone-like HAp. The coating of PS plates with HAp promoted the adhesion of mesenchymal stem cells (MSCs) and maintained cell growth that was as fast as that on tissue culture-treated PS (TCPS) plates. Osteogenic differentiation was greater, whereas adipogenic and chondrogenic differentiation was less in the culture on HAp-coated PS plates than in that on TCPS plates. The present method is useful for preparing HAp-coated PS plates at clean benches without the need for any expensive apparatus. HAp coated on PS plates by this method was a bone-like apatite with high bioactivity; therefore, the present HAp-coated PS plates are promising materials for assays of bone-related cells in the bone remodeling process.

  6. Effects of Lactic Acid Bacteria Inoculated Fermentation on Pickled Cucumbers

    OpenAIRE

    2013-01-01

    The aim of this study was to determine the effects of Lactic Acid Bacteria (LAB) fermentation on the texture and organic acid of pickled cucumbers. Texture and sensory evaluation as well as a microscopic observation were performed to study the textural differences among fresh cucumber, Spontaneous fermentation (SF) cucumber and LAB Inoculating Fermentation (LABIF) cucumber. Accumulation of seven organic acids i.e., oxalic, tartaric, malic, lactic, acetic, citric and succinic acid during cucum...

  7. Contralateral compartment syndrome inoculated by invasive group A streptococcus

    Directory of Open Access Journals (Sweden)

    Huiwen Chen

    2016-10-01

    Full Text Available Compartment syndrome is a rare but a well-documented complication in patients with trauma-induced group A streptococcus infection. Here, we present a case of a male who developed compartment syndrome on the left lower extremity after an injury inoculated by group A streptococcus on the right lower extremity. The patient was resuscitated with antibiotics, urgent fasciotomy, and immunoglobulin. The patient was eventually transferred to a burn center for further care.

  8. Inoculation with Azospirillum, associated with nitrogen fertilization in maize

    Directory of Open Access Journals (Sweden)

    Maria do Carmo Lana

    2012-06-01

    Full Text Available The biological nitrogen fixation is an alternative to supply the nitrogen needed for maize. The objective of this study was to evaluate the development and yield of maize in response to inoculation with Azospirillum associated with nitrogen fertilization. We conducted two field experiments in the summer harvest, the first in the 2000/2001 crop year in the region of Marechal Cândido Rondon, under conventional tillage, and second in the 2002/2003 agricultural year in the region of Cascavel, under no tillage. The experimental design in both experiments was a randomized complete block, with four replications, 2x2x2 factorial, with two levels of nitrogen at sowing (zero and 20 kg ha-1, two levels of inoculum (zero and 200 g ha-1 and two levels of nitrogen in topdressing (zero and 100 kg ha-1. There was evaluated the height of ear insertion, total plant height, leaf N content, shoot dry biomass and grain yield. The height of ear insertion and total plant height were not influenced by the factors under study. Nitrogen fertilization at sowing increased the leaf N content, causing the opposite effect when combined with inoculation. Inoculation with Azospirillum in the absence of nitrogen, provide productivity increases of 15.4% and 7.4% for 2000/2001 and 2002/2003 crops, respectively. The inoculation provided productivity similar to that obtained with 100 kg ha-1 in topdressing in crop 2000/2001, while in association with the topdressing, reduced productivity and shoot dry biomass in crop 2002/2003.

  9. Stress inoculation training: a case study in gymnastics.

    OpenAIRE

    Mace, R; Eastman, C.; Carroll, D.

    1986-01-01

    A young female gymnast of regional squad potential had ceased to make progress when she resumed training after a series of injuries and was given stress inoculation training to help her to regain her form. Preliminary interviews revealed that she had developed a number of negative self-statements and images which, it was hypothesised, may have been contributing towards her lack of progress. In order to replace these with positive self-statements and images a treatment programme of eight train...

  10. Two stages kinetics of municipal solid waste inoculation composting processes

    Institute of Scientific and Technical Information of China (English)

    XI Bei-dou1; HUANG Guo-he; QIN Xiao-sheng; LIU Hong-liang

    2004-01-01

    In order to understand the key mechanisms of the composting processes, the municipal solid waste(MSW) composting processes were divided into two stages, and the characteristics of typical experimental scenarios from the viewpoint of microbial kinetics was analyzed. Through experimentation with advanced composting reactor under controlled composting conditions, several equations were worked out to simulate the degradation rate of the substrate. The equations showed that the degradation rate was controlled by concentration of microbes in the first stage. The degradation rates of substrates of inoculation Run A, B, C and Control composting systems were 13.61 g/(kg·h), 13.08 g/(kg·h), 15.671 g/(kg·h), and 10.5 g/(kg·h), respectively. The value of Run C is around 1.5 times higher than that of Control system. The decomposition rate of the second stage is controlled by concentration of substrate. Although the organic matter decomposition rates were similar to all Runs, inoculation could reduce the values of the half velocity coefficient and could be more efficient to make the composting stable. Particularly. For Run C, the decomposition rate is high in the first stage, and is low in the second stage. The results indicated that the inoculation was efficient for the composting processes.

  11. Successive site translocating inoculation potentiates DNA/recombinant vaccinia vaccination.

    Science.gov (United States)

    Ren, Yanqin; Wang, Na; Hu, Weiguo; Zhang, Xiaoyan; Xu, Jianqing; Wan, Yanmin

    2015-12-15

    DNA vaccines have advantages over traditional vaccine modalities; however the relatively low immunogenicity restrains its translation into clinical use. Further optimizations are needed to get the immunogenicity of DNA vaccine closer to the level required for human use. Here we show that intramuscularly inoculating into a different limb each time significantly improves the immunogenicities of both DNA and recombinant vaccinia vaccines during multiple vaccinations, compared to repeated vaccination on the same limb. We term this strategy successive site translocating inoculation (SSTI). SSTI could work in synergy with genetic adjuvant and DNA prime-recombinant vaccinia boost regimen. By comparing in vivo antigen expression, we found that SSTI avoided the specific inhibition of in vivo antigen expression, which was observed in the limbs being repeatedly inoculated. Employing in vivo T cell depletion and passive IgG transfer, we delineated that the inhibition was not mediated by CD8(+) T cells but by specific antibodies. Finally, by using C3(-/-) mouse model and in vivo NK cells depletion, we identified that specific antibodies negatively regulated the in vivo antigen expression primarily in a complement depended way.

  12. Inoculation of maize with Azospirillum brasilense in the seed furrow

    Directory of Open Access Journals (Sweden)

    Tâmara Prado de Morais

    2016-06-01

    Full Text Available ABSTRACT Several studies addressing the inoculation of cereals with diazotrophic microorganisms can be found in the literature. However, in many experiments, investigators have overlooked the feasibility of applying these microorganisms to the furrow together with the seed, and the effect of bacterial concentration on phytostimulation. The aim of this work was to evaluate the effect of doses of an inoculant based on Azospirillum brasilense, applied to the seed furrow when planting maize, combined with different doses of nitrogen fertiliser. The experiment was carried out in the field, in soil of the cerrado region of Brazil. An experimental design of randomised blocks in bands was adopted, comprising nitrogen (40, 100, 200 and 300 kg ha-1 and doses of an A. brasilense-based liquid inoculant applied to the seed furrow (0, 100, 200, 300 and 400 mL ha-1. The dose of 200 mL ha-1Azospirillum was noteworthy for grain production. This is the first report of the effective application of Azospirillum in the seed furrow when planting maize in the cerrado region of Brazil.

  13. [HBsAg expression, anti-HBs induction and pathological observation in the mice inoculated with DNA vaccine against hepatitis B].

    Science.gov (United States)

    Zhao, L; Qin, S; Tang, H

    1999-03-01

    To study if special humoral immunological response can be induced by DNA vaccine against Hepatitis B (NV-HB/s) and its mechanism. The Balb/C mice inoculated with NV-HB/s by intramuscular injection were detected for HBsAg(the expressed product of NV-HB/s) by ABC immunohistochemistry or ELISA, and for anti-HBs by ELISA and also performed the routine pathological examination. After inoculated with NV-HB/s, HBsAg could be detected from mice muscle tissue samples of injected sites one week later and almost kept positive(75% and even more) up to 6th month, but was undetectable from all the serum samples during this period; anti-HBs could be detected from serum samples of some mice two weeks later and of all the mice detected one month later, it still kept positive at 6th month while the mice were sacrificed. Under microscope, only non-specific inflammation was found in the muscle tissues of injected sites and could completely recover within 4 weeks, no matter whether the mice were inoculated with NV-HBs, or with traditional HB vaccine derived from blood, or even with PBS as control. DNA vaccine against Hepatitis B (NV-HB/s) can successfully express HBsAg in the muscle tissue of mice by intramuscular inoculation and subsequently induce anti-HBs in vitro.

  14. Very long chain alkylresorcinols accumulate in the intracuticular wax of rye (Secale cereale L.) leaves near the tissue surface.

    Science.gov (United States)

    Ji, Xiufeng; Jetter, Reinhard

    2008-03-01

    Alkylresorcinols (ARs) are bioactive compounds occurring in many members of the Poaceae, likely at or near the surface of various organs. Here, we investigated AR localization within the cuticular wax layers of rye (Secale cereale) leaves. The total wax mixture from both sides of the leaves was found to contain primary alcohols (71%), alkyl esters (11%), aldehydes (5%), and small amounts (synthetic standard of nonadecylresorcinol. The alkyl side chains of the wax ARs contained odd numbers of carbons ranging from C19 to C27, with a prevalence of C21, C23 and C25. Waxes from both sides of the leaf, analyzed separately in a second experiment, comprised the same compound classes in similar relative amounts and with similar homolog patterns. Finally, the epicuticular and intracuticular wax layers were sampled separately from the abaxial side of the leaf. While ARs accounted for 2% of the intracuticular wax, they were not detectable in the epicuticular wax. The intracuticular wax was also slightly enriched in steroids, whereas the epicuticular layer contained more primary alcohols. All other wax constituents were distributed evenly between both wax layers.

  15. Effect of gamma irradiation on commercial eggs experimentally inoculated with Salmonella enteritidis

    Science.gov (United States)

    Tellez, I. G.; Trejo, R. M.; Sanchez, R. E.; Ceniceros, R. M.; Luna, Q. P.; Zazua, P.; Hargis, B. M.

    1995-02-01

    Using intact, fresh shell eggs, inoculated with 10 8 colony-forming units (cfu) of S. enteritidis, the effect of three doses of gamma irradiation on bacteriologic population and physical characteristics (Haugh units and yolk color) of the eggs was determinated. Penetration test area was picked at random just off the air cell of each egg. Aluminum cylinders were attached to the egg surface with a rim of molten paraffin, and 10 8S. enteritidis was then applied to inoculate the egg. Eggs were then irradiated within 2 hours using a Cobalt-60 gamma source at either 1, 2, or 3 kGy. A second set of inoculated, non-irradiated was used as controls. Following irradiation, eggs were maintained at 4°C for 42 hours prior culture. Irradiation with 1 kGy resulted in a significant (P < .05), 3.9 log reduction in detectable S. enteritidis in the shell and a higly significant (P < .025) 95% reduction in detectable S. enteritidis in the internal shell membranes. Irradiation of eggs with either 2 or 3 kGy reduced bacterial contamination to non-detectable levels in both the shell and internal membranes. However, irradiation at either 1, 2 or 3 kGy resulted in a significant (P <- .05) decrease (approximately 50%) in Haugh units. Additionally, irradiation of intact shell eggs at 2 or 3 Kgy significantly (P ≤ .05) reduced yolk color regardless of the level of irradiation exposure implemented. This data indicates that gamma irradiation of intact raw eggs is effective in reducing (1 kGy) or eliminating (2 or more kGy) S. enteritidis contamination. However, each of the levels of irradiation used in the present experiments caused marked reduction of selected measures of egg quality.

  16. In vitro and in vivo inoculation of four endophytic bacteria on Lycopersicon esculentum.

    Science.gov (United States)

    Botta, Anna Lucia; Santacecilia, Alessandra; Ercole, Claudia; Cacchio, Paola; Del Gallo, Maddalena

    2013-09-25

    Four bacteria selected on the basis of their capability of fixing atmospheric nitrogen, stimulating plant-growth, and protecting the host plant from pathogens - Azospirillum brasilense, Gluconacetobacter diazotrophicus, Herbaspirillum seropedicae, Burkholderia ambifaria - were inoculated on tomato seeds either singularly, in couple and in a four bacteria mixer. Aim of this research was to evaluate: (1) effect of single and mixed cultures on the inoculated plant - plant growth, dry weight, root length and surface, number of leaves, among others; (2) colonization and interactions of the bacteria inside the host plant; (3) localization inside the host of single bacterial strains marked with the gusA reporter gene. The results obtained indicate that all selected microbial strains have colonized Lycopersicon esculentum but in a different way, depending on the single species. A. brasilense, G. diazotrophicus inoculated in vitro singularly and together were the best plant colonizers. In vivo essays, instead, B. ambifaria and the four-bacteria mixer gave the best results. It was possible to localize both A. brasilense and H. seropedicae inside the plant by the gusA reporter gene. The bacterial strains occur along the root axis from the apical zone until to the basal stem, on the shoot from the base up to the leaves. The four bacteria actively colonize tomato seeds and establish an endophytic community inside the plant. This review gives new information about colonization processes, in particular how bacteria interact with plants and whether they are likely to establish themselves in the plant environment after field application as biofertilizers or biocontrol agents.

  17. Top-Down and Bottom-Up Identification of Proteins by Liquid Extraction Surface Analysis Mass Spectrometry of Healthy and Diseased Human Liver Tissue

    Science.gov (United States)

    Sarsby, Joscelyn; Martin, Nicholas J.; Lalor, Patricia F.; Bunch, Josephine; Cooper, Helen J.

    2014-09-01

    Liquid extraction surface analysis mass spectrometry (LESA MS) has the potential to become a useful tool in the spatially-resolved profiling of proteins in substrates. Here, the approach has been applied to the analysis of thin tissue sections from human liver. The aim was to determine whether LESA MS was a suitable approach for the detection of protein biomarkers of nonalcoholic liver disease (nonalcoholic steatohepatitis, NASH), with a view to the eventual development of LESA MS for imaging NASH pathology. Two approaches were considered. In the first, endogenous proteins were extracted from liver tissue sections by LESA, subjected to automated trypsin digestion, and the resulting peptide mixture was analyzed by liquid chromatography tandem mass spectrometry (LC-MS/MS) (bottom-up approach). In the second (top-down approach), endogenous proteins were extracted by LESA, and analyzed intact. Selected protein ions were subjected to collision-induced dissociation (CID) and/or electron transfer dissociation (ETD) mass spectrometry. The bottom-up approach resulted in the identification of over 500 proteins; however identification of key protein biomarkers, liver fatty acid binding protein (FABP1), and its variant (Thr→Ala, position 94), was unreliable and irreproducible. Top-down LESA MS analysis of healthy and diseased liver tissue revealed peaks corresponding to multiple (~15-25) proteins. MS/MS of four of these proteins identified them as FABP1, its variant, α-hemoglobin, and 10 kDa heat shock protein. The reliable identification of FABP1 and its variant by top-down LESA MS suggests that the approach may be suitable for imaging NASH pathology in sections from liver biopsies.

  18. Top-down and bottom-up identification of proteins by liquid extraction surface analysis mass spectrometry of healthy and diseased human liver tissue.

    Science.gov (United States)

    Sarsby, Joscelyn; Martin, Nicholas J; Lalor, Patricia F; Bunch, Josephine; Cooper, Helen J

    2014-11-01

    Liquid extraction surface analysis mass spectrometry (LESA MS) has the potential to become a useful tool in the spatially-resolved profiling of proteins in substrates. Here, the approach has been applied to the analysis of thin tissue sections from human liver. The aim was to determine whether LESA MS was a suitable approach for the detection of protein biomarkers of nonalcoholic liver disease (nonalcoholic steatohepatitis, NASH), with a view to the eventual development of LESA MS for imaging NASH pathology. Two approaches were considered. In the first, endogenous proteins were extracted from liver tissue sections by LESA, subjected to automated trypsin digestion, and the resulting peptide mixture was analyzed by liquid chromatography tandem mass spectrometry (LC-MS/MS) (bottom-up approach). In the second (top-down approach), endogenous proteins were extracted by LESA, and analyzed intact. Selected protein ions were subjected to collision-induced dissociation (CID) and/or electron transfer dissociation (ETD) mass spectrometry. The bottom-up approach resulted in the identification of over 500 proteins; however identification of key protein biomarkers, liver fatty acid binding protein (FABP1), and its variant (Thr→Ala, position 94), was unreliable and irreproducible. Top-down LESA MS analysis of healthy and diseased liver tissue revealed peaks corresponding to multiple (~15-25) proteins. MS/MS of four of these proteins identified them as FABP1, its variant, α-hemoglobin, and 10 kDa heat shock protein. The reliable identification of FABP1 and its variant by top-down LESA MS suggests that the approach may be suitable for imaging NASH pathology in sections from liver biopsies.

  19. Benefits of inoculation with azotobacter in the growth and production of tomato and peppers

    Directory of Open Access Journals (Sweden)

    Jarak Mirjana N.

    2010-01-01

    Full Text Available The aim of this research was to investigate the effects of Azotobacter chroococcum in tomato and pepper growth and production by using two types of inoculation - seed inoculation and seedling inoculation. The effect of inoculation was observed thirty days after sowing, thirty days after transplanting, and in the phase of technological maturity. The following were measured: height of the plants, dry matter of the plants and number and the weight of the fruits. Inoculation had a positive effect on these in both plants. With tomato, better results were achieved when seedlings were inoculated. With pepper, the length of the plant and the dry matter were greater with seedling inoculation, whereas the number and the weight of the fruits were greater with seed inoculation.

  20. Development of the "Three-step MACS": a novel strategy for isolating rare cell populations in the absence of known cell surface markers from complex animal tissue.

    Science.gov (United States)

    Lee, Mathia Y; Lufkin, Thomas

    2012-07-01

    To circumvent the difficulty of isolating specific cell populations by MACS from dissociated complex animal tissue, when their proportions reached levels similar to that of the background, we developed the "Three-step MACS" strategy. Cells of interest are defined by their expression of a particular gene(s) of interest rather by than their natural cell surface markers or size. A two-component transgenic cell surface protein, for two sequential rounds of MACS, is expressed under the promoter control of the endogenous gene of interest by means of gene targeting and the generation of transgenic tissue. An initial step to remove dead cells is also used. Here, we describe proof-of-concept experiments, using the biotin acceptor peptide (BAP)-low-affinity nerve growth factor receptor as the two-component protein. The first component, the BAP, can be biotinylated in specific subsets of cells expressing a particular gene by expressing the biotinylating enzyme, hBirA = humanized BirA (hBirA), under the promoter control of another gene defining the specific subpopulation. We showed that a rare population of cells (1.1% of the 13.5 days postcoital mouse embryo) could be enriched to a sufficiently high purity (84.4%). From another sample with 0.1% of our cells of interest, we achieved a 40.3% pure sample. The low cost, speed, and technical ease of the Three-step MACS also make it scalable and hence, an ideal method for preparing sufficient quantities of biological samples for sensitive, high-throughput assays.

  1. Direct plantlet inoculation with soil or insect-associated fungi may control cabbage root fly maggots.

    Science.gov (United States)

    Razinger, Jaka; Lutz, Matthias; Schroers, Hans-Josef; Palmisano, Marilena; Wohler, Christian; Urek, Gregor; Grunder, Jürg

    2014-07-01

    A potential Delia radicum biological control strategy involving cauliflower plantlet inoculation with various fungi was investigated in a series of laboratory and glasshouse experiments. In addition to entomopathogenic fungi, fungi with a high rhizosphere competence and fungi with the ability to survive as saprotrophs in soil were tested. The following fungal species were evaluated in the experiments: Trichoderma atroviride, T. koningiopsis, T. gamsii, Beauveria bassiana, Metharhizium anisopliae, M. brunneum and Clonostachys solani. A commercial carbosulfan-based insecticide was used as a positive control. Additionally, two commercial products, one based on B. bassiana (Naturalis) and one on Bacillus thuringiensis (Delfin) were used as reference biocontrol agents. The aims were (i) to assess the pathogenicity of the selected fungal isolates to Delia radicum, (ii) to evaluate the fungal isolates' rhizosphere competence, with the emphasis on the persistence of the original inoculum on the growing roots, (iii) to assess possible endophytic plant tissue colonization, and (iv) to evaluate potential plant growth stimulating effects of the added inoculi. Significant pathogenicity of tested fungi against Delia radicum was confirmed in in vitro and glasshouse experiments. All tested fungi persisted on cauliflower rhizoplane. More importantly, the added fungi were found on thoroughly washed roots outside the original point of inoculation. This provided us with evidence that our tested fungi could be transferred via or grow with the elongating roots. In addition to colonizing the rhizoplane, some fungi were found inside the plant root or stem tissue, thus exhibiting endophytic characteristics. The importance of fungal ecology as a criterion in appropriate biological control agent selection is discussed. Copyright © 2014 Elsevier Inc. All rights reserved.

  2. Effects of the inoculation of Burkholderia vietnamensis and related endophytic diazotrophic bacteria on grain yield of rice.

    Science.gov (United States)

    Govindarajan, Munusamy; Balandreau, Jacques; Kwon, Soon-Wo; Weon, Hang-Yeon; Lakshminarasimhan, Cunthipuram

    2008-01-01

    During a survey of endophytic diazotrophic bacteria associated with different rice varieties in Tamilnadu, some "endophytes" were obtained. Thirteen bacterial isolates from surface-sterilized roots and shoots were obtained in pure culture, which produced indole acetic acid (IAA) and reduced acetylene to ethylene. Polymerase chain reaction (PCR) amplification confirmed the presence of nif-H gene in all the isolates. Morphological, biochemical, and molecular characteristics indicated that all of them belonged to the genus Burkholderia One of them, MGK3, was consistently more active in reducing acetylene, and 16S rDNA sequences of isolate MGK3 confirmed its identification as Burkholderia vietnamiensis. Colonization of rice root was confirmed by strain MGK3 marked with gusA gene. The inoculated roots showed a blue color, which was most intense at the points of lateral root emergence and at the root tip. Transverse sections of roots, 15 days after inoculation, revealed beta-glucuronidase (GUS) activity within many of the cortical intercellular spaces next to the stele and within the aerenchyma. Nitrogen fixation was quantified by using (15)N isotope dilution method with two different cultivars grown in pot and field experiments. Higher nitrogen fixation was observed in variety Ponni than in ADT-43, where nearly 42% (field) and 40% (pot) of the nitrogen was derived from the atmosphere (% Ndfa). Isolate MGK3 was used to inoculate rice seedlings in a comparison with four other diazotrophs, viz., Gluconacetobacter diazotrophicus LMG7603, Herbaspirillum seropedicae LMG6513, Azospirillum lipoferum 4B LMG4348, and B. vietnamiensis LMG10929. They were used to conduct two pot and four field inoculation experiments. MGK3 alone, and combined with other diazotrophs, performed best under both pot and field conditions: combined inoculation produced yield increases between 9.5 and 23.6%, while MGK3 alone increased yield by 5.6 to 12.16% over the uninoculated control treatment.

  3. Highly structured and surface modified poly(epsilon-caprolactone) scaffolds derived from co-continuous polymer blends for bone tissue engineering

    Science.gov (United States)

    Mehr, Nima Ghavidel

    L) surface modification has never been evaluated before. This part of the study tests the hypothesis that in vitro osteogenesis can be achieved in 3D PCL scaffolds with fully interconnected pores of 84 im or 141 im average diameter and biomineralization can be enhanced when pore surfaces are coated with chitosan adsorbed to LbL deposited polyelectrolytes. In order to reduce the errors originating from cell infiltration inefficiencies, the most competent cell seeding protocol has to be defined. Among classical cell seeding at 37°C, 2-step seeding at 37°C and cold seeding at 4°C in a medium containing 2% FBS, the last strategy proved to yield the best population of freshly trypsinized hBMSCs at all depths of the 1mm-thick scaffolds. hBMSCs cold-seeded in PCL scaffolds with or without an LbL-chitosan coating were cultured for 10 days in proliferation medium, followed by 21 days in osteogenic medium. At day 2, MSCs formed sparse monolayers with rounded cell morphologies with thin filopodia anchored to the unmodified PCL, as compared to more spread cells on chitosan-coated pore surfaces. At day 10, cells proliferated as an external layer, and migrated onto secreted collagen networks that filled the interpore spaces of all scaffolds, but only adhered to chitosan-coated pore surfaces. At day 31, similar levels of tissue formed in scaffolds with and without chitosan, but more tissue was deposited in the outer pores than the inner pores. Furthermore, more biomineralized matrix was observed in the inner 84 im chitosan-coated pores (p<0.05). In the PCL-only samples, haphazard mineral deposits were observed in highly colonized outer layers and in the inner 141 im pores. MSCs cultured on chitosan-coated 2D control surfaces show higher alkaline phosphatase staining but negligible mineralization. This study showed that hBMSCs survive, proliferate, and attach to fibrotic matrix rather than the PCL-only scaffold pore surfaces. LbL-chitosan-coated scaffolds showed more biomineralization in

  4. Limited amplification of chronic wasting disease prions in the peripheral tissues of intracerebrally inoculated cattle

    Science.gov (United States)

    Chronic wasting disease (CWD) is a fatal neurodegenerative disease, classified as a prion disease or transmissible spongiform encephalopathy (TSE) similar to bovine spongiform encephalopathy (BSE). Cervids affected by CWD accumulate an abnormal protease resistant prion protein throughout the central...

  5. Inoculation of weaned pigs with E. coli reduces depots of vitamin E

    DEFF Research Database (Denmark)

    Lauridsen, Charlotte; Vestergaard, Ellen-Margrethe; Højsgaard, Søren

    2011-01-01

    60 mg all-rac-α-tocopheryl acetate) and E. coli 0 149 inoculation (inoculation of 1 × 108 CFU on day 2 and 3 after weaning versus inoculation of vehicle). The pigs were housed individually during the experiment which lasted for 10 days from weaning at 7 weeks of age. Blood was sampled on day 1 (day...

  6. Experimental Inoculation of Egyptian Rousette Bats (Rousettus aegyptiacus with Viruses of the Ebolavirus and Marburgvirus Genera

    Directory of Open Access Journals (Sweden)

    Megan E.B. Jones

    2015-06-01

    Full Text Available The Egyptian rousette bat (Rousettus aegyptiacus is a natural reservoir for marburgviruses and a consistent source of virus spillover to humans. Cumulative evidence suggests various bat species may also transmit ebolaviruses. We investigated the susceptibility of Egyptian rousettes to each of the five known ebolaviruses (Sudan, Ebola, Bundibugyo, Taï Forest, and Reston, and compared findings with Marburg virus. In a pilot study, groups of four juvenile bats were inoculated with one of the ebolaviruses or Marburg virus. In ebolavirus groups, viral RNA tissue distribution was limited, and no bat became viremic. Sudan viral RNA was slightly more widespread, spurring a second, 15-day Sudan virus serial euthanasia study. Low levels of Sudan viral RNA disseminated to multiple tissues at early time points, but there was no viremia or shedding. In contrast, Marburg virus RNA was widely disseminated, with viremia, oral and rectal shedding, and antigen in spleen and liver. This is the first experimental infection study comparing tissue tropism, viral shedding, and clinical and pathologic effects of six different filoviruses in the Egyptian rousette, a known marburgvirus reservoir. Our results suggest Egyptian rousettes are unlikely sources for ebolaviruses in nature, and support a possible single filovirus—single reservoir host relationship.

  7. Automated Liquid Microjunction Surface Sampling-HPLC-MS/MS Analysis of Drugs and Metabolites in Whole-Body Thin Tissue Sections

    Energy Technology Data Exchange (ETDEWEB)

    Kertesz, Vilmos [ORNL; Van Berkel, Gary J [ORNL

    2013-01-01

    A fully automated liquid extraction-based surface sampling system utilizing a commercially available autosampler coupled to high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) detection is reported. Discrete spots selected for droplet-based sampling and automated sample queue generation for both the autosampler and MS were enabled by using in-house developed software. In addition, co-registration of spatially resolved sampling position and HPLC-MS information to generate heatmaps of compounds monitored for subsequent data analysis was also available in the software. The system was evaluated with whole-body thin tissue sections from propranolol dosed rat. The hands-free operation of the system was demonstrated by creating heatmaps of the parent drug and its hydroxypropranolol glucuronide metabolites with 1 mm resolution in the areas of interest. The sample throughput was approximately 5 min/sample defined by the time needed for chromatographic separation. The spatial distributions of both the drug and its metabolites were consistent with previous studies employing other liquid extraction-based surface sampling methodologies.

  8. Atmospheric pressure plasma treatment of black peppercorns inoculated with Salmonella and held under controlled storage.

    Science.gov (United States)

    Sun, Shengqian; Anderson, Nathan M; Keller, Susanne

    2014-12-01

    Spices, including black pepper, are a source of microbial contamination and have been linked to outbreaks of salmonellosis when added to products that undergo no further processing. Traditional thermal processing employed to reduce microbial contamination can lead to losses of heat-sensitive compounds. Thus, alternative processes such as atmospheric pressure plasma (APP) are desirable. The purpose of this research was to determine the efficacy of APP in the destruction of Salmonella inoculated on the surface of peppercorns. Secondarily, we examined the effect of storage on the subsequent inactivation of Salmonella on the surfaces of black peppercorns by APP. Black peppercorns inoculated with a cocktail of Salmonella enterica serotypes Oranienburg, Tennessee, Anatum, and Enteritidis were stored at 25 °C, 33% relative humidity (RH); 25 °C, 97% RH; and, 37 °C, 33% RH for 10 d and additionally at 25 °C, 33% RH for 1 and 30 d then treated with APP. Results showed that Salmonella populations decreased significantly (P 0.05). Approximately a 4.5- to 5.5-log10 reduction in population was achieved after 60 to 80 s treatment. A combination of treatments, storage and 80 s of plasma, may achieve a total reduction on the order of 7-log10 CFU/g. These findings support the potential of APP to decontaminate Salmonella on the surfaces of black peppercorns and other dry foods and illustrate that a multiple hurdle approach may prove effective for achieving significant reductions of Salmonella in many low-moisture foods.

  9. Intranasal inoculation of white-tailed deer (Odocoileus virginianus with lyophilized chronic wasting disease prion particulate complexed to montmorillonite clay.

    Directory of Open Access Journals (Sweden)

    Tracy A Nichols

    Full Text Available Chronic wasting disease (CWD, the only known prion disease endemic in wildlife, is a persistent problem in both wild and captive North American cervid populations. This disease continues to spread and cases are found in new areas each year. Indirect transmission can occur via the environment and is thought to occur by the oral and/or intranasal route. Oral transmission has been experimentally demonstrated and although intranasal transmission has been postulated, it has not been tested in a natural host until recently. Prions have been shown to adsorb strongly to clay particles and upon oral inoculation the prion/clay combination exhibits increased infectivity in rodent models. Deer and elk undoubtedly and chronically inhale dust particles routinely while living in the landscape while foraging and rutting. We therefore hypothesized that dust represents a viable vehicle for intranasal CWD prion exposure. To test this hypothesis, CWD-positive brain homogenate was mixed with montmorillonite clay (Mte, lyophilized, pulverized and inoculated intranasally into white-tailed deer once a week for 6 weeks. Deer were euthanized at 95, 105, 120 and 175 days post final inoculation and tissues examined for CWD-associated prion proteins by immunohistochemistry. Our results demonstrate that CWD can be efficiently transmitted utilizing Mte particles as a prion carrier and intranasal exposure.

  10. Growth and histamine formation of Morganella morganii in determining the safety and quality of inoculated and uninoculated bluefish (Pomatomus saltatrix).

    Science.gov (United States)

    Lorca, T A; Gingerich, T M; Pierson, M D; Flick, G J; Hackney, C R; Sumner, S S

    2001-12-01

    The objective of this study was to determine the effect of normal microflora and Morganella morganii on histamine formation and olfactory acceptability in raw bluefish under controlled storage conditions. Fillets inoculated with and without M. morganii were stored at 5, 10, and 15 degrees C for 7 days. Microbial isolates from surface swabs were identified and screened for histidine decarboxylase activity. Olfactory acceptance was performed by an informal sensory panel. Histamine levels were quantified using high-performance liquid chromatography and fluorescence detection. While olfactory acceptance decreased, histamine concentration and bacterial counts increased. Storage temperature had a significant effect on histamine levels, bacterial counts, and olfactory acceptance of the bluefish. Inoculation with M. morganii had a positive significant effect on histamine formation for bluefish held at 10 and 15 degrees C (P < 0.0001). The results of the study will serve in supporting U.S. Food and Drug Administration (FDA) regulations regarding guidance and hazard levels of histamine in fresh bluefish.

  11. "DIFERENTIAION OF LEISHMANIA TROPICA MAJOR FROM LEISHMANIA TROPICA MINOR BY INOCULATION TO LABORATORY ANIMALS "

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    A.Nadim

    1973-08-01

    Full Text Available The subcutaneous inoculation of 16 white mice and 10 golden hamsters with L.tropica major resulted in the development of leishmanial lesions in all, becoming generalized in some.Two of 10 hamsters inoculated intraperitoneally with this parasite developed generalized lesions. On the other hand, the inoculation of L. tropica minor to 128 mice subcutaneously and 170 mice intraperitoneally did not cause any leishmanial lesions , while 22 out of 47 hamsters inoculated with this parasite showed lesions at the site of inoculation.

  12. Effects of Homofermentative Lactic Acid Bacterial Inoculants on the Fermentation and Aerobic Stability Characteristics of Low Dry Matter Corn Silages

    OpenAIRE

    SUCU, Ekin; FİLYA, İsmail

    2014-01-01

    This study was carried out to determine the effects of homofermentative lactic acid bacterial inoculants on the fermentation and aerobic stability characteristics of low dry matter corn silages. Corn was harvested at the milk stage. Inoculant-1188 (Pioneer®, USA; Inoculant A) and Maize-All (Alltech, UK; Inoculant B) were used as homofermentative lactic acid bacterial inoculants. Inoculants were applied to silages 1.5 x 106 colony forming units/g levels. Silages with no additive served as co...

  13. Effectiveness of a sediment time critical removal action-PCB reduction in fish tissue, surface water, and sediment via wet excavation.

    Science.gov (United States)

    Santini, Andrew D; King, Todd; Krawczyk, Keith; Kern, John W

    2015-01-01

    Documenting successful remediation of polychlorinated biphenyl (PCB)-contaminated sediments is limited; potentially due to inadequate monitoring methods, complexities associated with the environment, and selected remedial techniques. At some sites, absence of appropriate baseline and postremoval monitoring limits proper evaluation of remedial efficacy. Accurate interpretation of interactions between media, space, time, species, lipid content, and remedial technique requires robust study design and data. This article presents baseline and postremoval data documenting reduced PCB concentrations in fish tissue, surface water, and sediment in response to the US Environmental Protection Agency (USEPA) time-critical removal action (TCRA) that was conducted at the former Bryant Mill Pond (BMP) on Portage Creek in Kalamazoo, Michigan. The BMP is part of an operable unit (OU) within the Allied Paper, Inc./Portage Creek/Kalamazoo River Superfund Site. PCBs discharged to the creek as a byproduct of carbonless copy paper recycling are the primary contaminant of concern. Paper waste residuals commonly appear as gray to light gray clays in river sediments and floodplain soils. The cleanup criterion was 10 mg/kg, with a residual PCB concentration goal of 1 mg/kg. Because the PCB-containing waste is (generally) associated with readily visible light gray clay, excavation of all visibly contaminated current or formerly impounded sediment served as a surrogate for the cleanup criteria and goal. Sediment was wet excavated and backfilled after diversion of the creek. After confirmation that PCB concentrations met cleanup criteria, the stream was diverted to the excavated side, and excavation and backfilling were completed. Overall, 146000 cubic yards of material including PCB-contaminated sediments were removed from the BMP. The long-term monitoring (LTM) program implemented by the Michigan Department of Environmental Quality (MDEQ), and historic data from a variety of sources

  14. Experimental induction of proventricular dilatation disease in cockatiels (Nymphicus hollandicus) inoculated with brain homogenates containing avian bornavirus 4.

    Science.gov (United States)

    Gancz, Ady Y; Kistler, Amy L; Greninger, Alexander L; Farnoushi, Yigal; Mechani, Sara; Perl, Shmuel; Berkowitz, Asaf; Perez, Noa; Clubb, Susan; DeRisi, Joseph L; Ganem, Don; Lublin, Avishai

    2009-07-09

    Proventricular dilatation disease (PDD) is a fatal disorder of psittacine birds worldwide. The disease is characterized by lymphoplasmacytic infiltration of the central and peripheral nervous systems, leading to gastrointestinal motility and/or central nervous system dysfunction. Recently, we detected a significant association between avian bornavirus (ABV) infection and clinical signs of PDD in psittacines. However, it remains unclear whether ABV infection actually causes PDD. To address this question, we examined the impact of ABV inoculation on the cockatiel (Nymphicus hollandicus). Five cockatiels were inoculated via multiple routes (intramuscular, intraocular, intranasal, and oral) with a brain homogenate derived from either a PDD(+) avian bornavirus 4 (ABV4) (+) case (n = 3 inoculees) or from a PDD(-) ABV(-) control (n = 2 inoculees). The control birds remained free of clinical or pathological signs of PDD, and tested ABV(-) by RT-PCR and immunohistochemistry (IHC). In contrast, all three cockatiels inoculated with ABV4(+) brain homogenate developed gross and microscopic PDD lesions, and two exhibited overt clinical signs. In numerous tissues, ABV RT-PCR and sequence analysis demonstrated the presence of ABV4 RNA nearly identical to that in the inoculum. ABV was detected in the central nervous system of the three ABV-inoculees by IHC. Pyrosequencing to investigate the viral flora in the ABV4(+) inoculum uncovered 7 unique reads sharing 73-100% nucleotide sequence identity with previously identified ABV sequences and 24 reads sharing 40-89% amino acid sequence identity with viruses in the Retroviridae and Astroviridae families. Of these candidate viral species, only ABV RNA was recovered from tissues of the inoculated birds. In this study, the clinical and pathological manifestations of PDD were induced by inoculation of cockatiels with brain homogenates containing avian bornavirus 4. By using high throughput pyrosequencing an in-depth view of the viral

  15. Experimental induction of proventricular dilatation disease in cockatiels (Nymphicus hollandicus inoculated with brain homogenates containing avian bornavirus 4

    Directory of Open Access Journals (Sweden)

    Berkowitz Asaf

    2009-07-01

    Full Text Available Abstract Background Proventricular dilatation disease (PDD is a fatal disorder of psittacine birds worldwide. The disease is characterized by lymphoplasmacytic infiltration of the central and peripheral nervous systems, leading to gastrointestinal motility and/or central nervous system dysfunction. Recently, we detected a significant association between avian bornavirus (ABV infection and clinical signs of PDD in psittacines. However, it remains unclear whether ABV infection actually causes PDD. To address this question, we examined the impact of ABV inoculation on the cockatiel (Nymphicus hollandicus. Results Five cockatiels were inoculated via multiple routes (intramuscular, intraocular, intranasal, and oral with a brain homogenate derived from either a PDD(+ avian bornavirus 4 (ABV4 (+ case (n = 3 inoculees or from a PDD(- ABV(- control (n = 2 inoculees. The control birds remained free of clinical or pathological signs of PDD, and tested ABV(- by RT-PCR and immunohistochemistry (IHC. In contrast, all three cockatiels inoculated with ABV4(+ brain homogenate developed gross and microscopic PDD lesions, and two exhibited overt clinical signs. In numerous tissues, ABV RT-PCR and sequence analysis demonstrated the presence of ABV4 RNA nearly identical to that in the inoculum. ABV was detected in the central nervous system of the three ABV-inoculees by IHC. Pyrosequencing to investigate the viral flora in the ABV4(+ inoculum uncovered 7 unique reads sharing 73–100% nucleotide sequence identity with previously identified ABV sequences and 24 reads sharing 40–89% amino acid sequence identity with viruses in the Retroviridae and Astroviridae families. Of these candidate viral species, only ABV RNA was recovered from tissues of the inoculated birds. Conclusion In this study, the clinical and pathological manifestations of PDD were induced by inoculation of cockatiels with brain homogenates containing avian bornavirus 4. By using high throughput

  16. THE EFFECTS OF INOCULANT LACTIC ACID BACTERIA ON THE FERMENTATION AND AEROBIC STABILITY OF SUNFLOWER SILAGE

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    Fisun Koc

    2009-12-01

    Full Text Available This study was carried out to determine the effects of actic acid bacterial inoculant on the fermentation and aerobic stability of sunflower silages. Sunflower was harvested at the milk stage. Inoculant-1174 (Pioneer®,USA was used as homofermentative lactic acid bacterial inoculant. Inoculant was applied 6.00 log10 cfu/g silage levels. Silages with no additive served as controls. After treatment, the chopped sunflower was ensiled in the PVC type laboratory silos. Three silos for each group were sampled for chemical and microbiological analysis on days 2, 4, 7, 14, 21, 28 and 56 after ensiling. At the end of the ensiling period, all silages were subjected to an aerobic stability test for 14 days. Neither inoculant improved the fermentation parameters of sunflower silages. At the end of the ensiling period, inoculant increased lactic acid bacteria (LAB and decreased yeast and mould numbers of silages. Inoculant treatment did not affect aerobic stability of silages.

  17. Anatomy and ultrastructure alterations of Leucaena leucocephala (Lam.) inoculated with mycorrhizal fungi in response to arsenic-contaminated soil

    Energy Technology Data Exchange (ETDEWEB)

    Schneider, Jerusa, E-mail: jerusaschneider@hotmail.com [Departamento de Ciência do Solo, Universidade Federal de Lavras (UFLA), PO Box 3037, Lavras, Minas Gerais, 37200-000 (Brazil); Labory, Claudia Regina Gontijo [Departamento de Fitopatologia, Universidade Federal de Lavras, PO Box 3037, Lavras, Minas Gerais, 37200-000 (Brazil); Rangel, Wesley Melo [Departamento de Ciência do Solo, Universidade Federal de Lavras (UFLA), PO Box 3037, Lavras, Minas Gerais, 37200-000 (Brazil); Alves, Eduardo [Departamento de Fitopatologia, Universidade Federal de Lavras, PO Box 3037, Lavras, Minas Gerais, 37200-000 (Brazil); Guilherme, Luiz Roberto Guimarães [Departamento de Ciência do Solo, Universidade Federal de Lavras (UFLA), PO Box 3037, Lavras, Minas Gerais, 37200-000 (Brazil)

    2013-11-15

    Highlights: ► Inoculation of L. leucocephala improved plant growth in high-As soils. ► Plants inoculated with Glomus clarum were less sensitive to As. ► Ultrastructural changes in leaves of L. leucocephala. ► Modified structures in intracellular spaces in plants inoculated with G. clarum. ► Cell disruption and stacking of root cell walls at high As concentrations. -- Abstract: Many studies demonstrate the potential application of arbuscular mycorrhizal fungi (AMF) for remediation purposes, but little is known on AMF potential to enhance plant tolerance to arsenic (As) and the mechanisms involved in this process. We carried anatomical and ultrastructural studies to examine this symbiotic association and the characteristics of shoots and roots of Leucaena leucocephala in As-amended soils (35 and 75 mg As dm{sup −3}). The experiment used 3 AMF isolates from uncontaminated soils: Acaulospora morrowiae, Glomus clarum, and Gigaspora albida; a mixed inoculum derived from combining these 3 isolates (named Mix AMF); and, 3 AMF isolates from As-contaminated areas: A. morrowiae, G. clarum and Paraglomus occultum. Phytotoxicity symptoms due to arsenic contamination appeared during plant growth, especially in treatments without AMF application. Inoculation with G. clarum and the mixture of species (A. morrowiae, G. albida, and G. clarum) resulted in better growth of L. leucocephala in soils with high As concentrations, as well as significant As removal from the soil, showing a potential for using AMF in phytoextraction. Light microscopy (LS), transmission (TEM) and scanning electron microscopies (SEM) studies showed the colonization of the AMF in plant tissues and damage in all treatments, with ultrastructural changes being observed in leaves and roots of L. leucocephala, especially with the addition of 75 mg dm{sup −3} of As.

  18. Use of inoculated lactic acid bacteria in fermenting sour cabbage

    Directory of Open Access Journals (Sweden)

    E. PETÄJÄ

    2008-12-01

    Full Text Available Fermentation of sour vegetables has to date occurred through the use of lactic acid bacteria (LAB naturally present in vegetables. The present article deals with preliminary studies on the effects of some LAB inocula (Lactobacillus alimentarius or Pediococcus pentosaceus on fermenting sour cabbage. The effect of LAB on yeast growth, a problem in sour vegetables, was also studied through the use of dual yeast and LAB inocula. The pH of cabbage juice decreased to levels under pH 4 during the first 10 days of fermentation, which is near the final values, pediococci decreasing the pH to the lowest values. The LAB count in inoculated cabbages increased by 0.5-2.0 log cfu (colony forming unit /g during the first 10 days of fermentation and thereafter decreased. Pediococci formed predominant part of microbial flora almost in all experimental batches. In cabbage challenged with yeasts, yeast counts rose only when the pH was < 3.5. Yeasts appeared almost regularly also in cabbages inoculated only with LAB. Pediococci fermented cabbage effectively decreasing the pH to lower levels than lactobacilli or natural LAB. However, too strong a decrease in pH may result in a decrease of LAB count which may subsequently lead to yeast growth. The yeast problem could not be solved with the LAB inocula used in our study.;

  19. Accumulation of Transcripts Abundance after Barley Inoculation with Cochliobolus sativus

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    Mohammad Imad Eddin Arabi

    2015-03-01

    Full Text Available Spot blotch caused by the hemibiotrophic pathogen Cochliobolus sativus has been the major yield-reducing factor for barley production during the last decade. Monitoring transcriptional reorganization triggered in response to this fungus is an essential first step for the functional analysis of genes involved in the process. To characterize the defense responses initiated by barley resistant and susceptible cultivars, a survey of transcript abundance at early time points of C. sativus inoculation was conducted. A notable number of transcripts exhibiting significant differential accumulations in the resistant and susceptible cultivars were detected compared to the non-inoculated controls. At the p-value of 0.0001, transcripts were divided into three general categories; defense, regulatory and unknown function, and the resistant cultivar had the greatest number of common transcripts at different time points. Quantities of differentially accumulated gene transcripts in both cultivars were identified at 24 h post infection, the approximate time when the pathogen changes trophic lifestyles. The unique and common accumulated transcripts might be of considerable interest for enhancing effective resistance to C. sativus.

  20. Dependency and Response of Apuleia leiocarpa to Inoculation with Different Species of Arbuscular Mycorrhizal Fungi

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    Joel Quintino de Oliveira Júnior

    Full Text Available ABSTRACT Inoculation with arbuscular mycorrhizal fungi (AMF is a strategy to improve the efficiency of forest plantations, reducing costs and increasing the survival of plant species. The objective of this study was to assess the response and mycorrhizal dependency of seedlings of the forest species Apuleia leiocarpa (Vogel J.F. Macbr to inoculation with AMF. The experiment was conducted in a completely randomized design using a 5 × 5 factorial arrangement with six replications. The treatments consisted of combinations of five P rates (0, 24, 71, 213, and 650 mg kg-1 with five types of inoculations with AMF (inoculation with the fungi Rhizophagus clarus, Gigaspora margarita, Dentiscutata heterogama, inoculation with an AMF mix of these three species, and a treatment without inoculation. The A. leiocarpa showed the highest biomass accumulations in inoculation with D. hetorogama combined with the P rates of 213 and 650 mg kg-1, and in the AMF mix combined with the P rates of 71, 213, and 650 kg-1. Biomass accumulation showed a linear, positive response to inoculation with D. heterogama combined with the different P rates, and a positive square root fit to inoculation with the AMF mix. The plants inoculated with G. margarita had no significant biomass accumulation. The plant species had a positive response to inoculation with R. clarus combined with the lowest P rates; however, it had a negative response to combination with the highest P rate (650 mg kg-1. The relative benefit of inoculation with these fungi was more than 100 % in most treatments, showing the high mycorrhizal dependency of A. leiocarpa and the nutritional benefit of AMF inoculation for this species. However, this response is dependent on the fungus species that colonize the plant roots. The best combination between fungus and P rate was inoculation with the AMF mix combined with the P rate of 71 mg kg-1.

  1. Lack of autologous tissue transmission of eosinophilic plaques in cats.

    Science.gov (United States)

    Moriello, K A; Kunkle, G; Miller, L M; Crowley, A

    1990-07-01

    Autologous tissue transmission of spontaneously developing feline eosinophilic plaques was attempted in 5 cats. Macerated tissue from the plaque was vigorously rubbed onto 2 scarified skin sites in each cat. The inoculated areas were observed daily for 30 days. During that time, no clinical or histologic evidence of transmission was found.

  2. Surface probe measurements of the elasticity of sectioned tissue, thin gels and polyelectrolyte multilayer films: Correlations between substrate stiffness and cell adhesion

    Science.gov (United States)

    Engler, Adam J.; Richert, Ludovic; Wong, Joyce Y.; Picart, Catherine; Discher, Dennis E.

    2004-10-01

    Surface probe measurements of the elasticity of thin film matrices as well as biological samples prove generally important to understanding cell attachment across such systems. To illustrate this, sectioned arteries were probed by atomic force microscopy (AFM) within the smooth muscle cell (SMC)-rich medial layer, yielding an apparent Young's modulus Emedia ˜ 5-8 kPa. Polyacrylamide gels with Egel spanning several-fold above and below this range were then cast 5-70 μm thick and coated with collagen: SMC spreading shows a hyperbolic dependence in projected cell area versus Egel. The modulus that gives half-max spreading is E1/2-spread ˜ 8-10 kPa, proving remarkably close to Emedia. More complex, layer-by-layer microfilms of poly( L-lysine)/hyaluronic acid were also tested and show equivalent trends of increased SMC spreading with increased stiffness. Adhesive spreading of cells thus seems to correlate broadly with the effective stiffness of synthetic materials and tissues.

  3. Efficiency of inoculant with Azospirillum brasilense on the growth and yield of second-harvest maize

    Directory of Open Access Journals (Sweden)

    Raoni Ribeiro Guedes Fonseca Costa

    2015-09-01

    Full Text Available The misuse of fertilizers has become a growing concern in terms of agricultural and environmental sustainability. One alternative to reduce the impact of chemical fertilizers is the use of inoculants containing nitrogen-fixing and plant growth promoting bacteria. This study aimed at evaluating the effects of an inoculant with Azospirillum brasilense on the growth and yield of secondharvest maize. The experimental design was randomized blocks with three replications, in a 3 × 5 factorial scheme. The treatments consisted of three inoculation methods (control without inoculation, seed inoculation and leaf-sprayed inoculation at the V4 stage and five cover N-fertilization doses (0 %, 25 %, 50 %, 75 % and 100 %, with the 100 % dose corresponding to 50 kg ha-1 of N as urea. Inoculated plants showed a greater growth for height (9.5 % and 16 %, stem dry biomass (49 % and 6.75 % and root dry biomass (123 % and 97 %, respectively for seed inoculation and spraying, as well as increases in ear size (8 %, chlorophyll content (4 %, 1,000-grain weight (12 % and grain yield (29 %, when compared to non-inoculated plants. The effects of N doses were only significant for 1,000-grain weight and grain yield (kg ha-1, with seed inoculation providing a higher yield for second-harvest maize.

  4. Agro-industrial waste materials and wastewater sludge for rhizobial inoculant production: a review.

    Science.gov (United States)

    Ben Rebah, F; Prévost, D; Yezza, A; Tyagi, R D

    2007-12-01

    Inoculating legumes with commercial rhizobial inoculants is a common agriculture practice. Generally, inoculants are sold in liquid or in solid forms (mixed with carrier). The production of inoculants involves a step in which a high number of cells are produced, followed by the product formulation. This process is largely governed by the cost related to the medium used for rhizobial growth and by the availability of a carrier source (peat) for production of solid inoculant. Some industrial and agricultural by-products (e.g. cheese whey, malt sprouts) contain growth factors such as nitrogen and carbon, which can support growth of rhizobia. Other agro-industrial wastes (e.g. plant compost, filtermud, fly-ash) can be used as a carrier for rhizobial inoculant. More recently, wastewater sludge, a worldwide recyclable waste, has shown good potential for inoculant production as a growth medium and as a carrier (dehydrated sludge). Sludge usually contains nutrient elements at concentrations sufficient to sustain rhizobial growth and heavy metals are usually below the recommended level. In some cases, growth conditions can be optimized by a sludge pre-treatment or by the addition of nutrients. Inoculants produced in wastewater sludge are efficient for nodulation and nitrogen fixation with legumes as compared to standard inoculants. This new approach described in this review offers a safe environmental alternative for both waste treatment/disposal and inoculant production.

  5. Effects of different inoculants on the microstructural characteristics of gray cast iron gg-25, hardness and useful life of tools

    Directory of Open Access Journals (Sweden)

    Diego Ruben Martin

    2015-10-01

    Full Text Available Current study evaluated the machinability characteristics of parts, microstructure and mechanical properties when three different inoculants (IM-22 with FeSi-Ba/Zr; G-20 and FeSi-Ba; IMSR 75 with FeSi-Sr were added in experiments carried out in a foundry. The research methodology was mailly based on the analysis of the machinability by the milling process of the specimens in gray cast iron GG-25, name according to DIN EN 1561.Evaluation of results is based on a thorough analysis of tool wear, surface finish, microstructural analysis, chemical composition and mechanical properties of the material. Results showed that among the studied inoculants strontium sulfide (SrS was thermodynamically more stable than the others, because it leds towards a more negative free energy change of Gibbs and therefore more favorable to the formation of nuclei having greater critical radius (rc, solidification with heterogeneous nucleation. Its inoculant was also more efficient in forming a more favorable microstructure, greater amounts of eutectic cells and, longer life of the insert when machined.

  6. Sampling by sponge wipe or skin excision for recovery of inoculated Salmonella and Campylobacter from defeathered broiler carcasses.

    Science.gov (United States)

    Berrang, Mark E; Cox, Nelson A; Oakley, Brian B

    2014-05-01

    Broilers may carry Salmonella and Campylobacter on inner and outer surfaces upon arrival at the slaughter plant, and carcasses can be further contaminated during commercial processing. A sensitive, nondestructive, repeatable sampling method would be useful to test carcasses for levels of bacteria before and after specific processing steps to measure either contamination or efficacy of intervention techniques. Blending of excised skin is accepted as an effective sampling method but requires damage to the carcass; this makes repeated measurements on the same carcass difficult. Herein we compare sponge sampling to skin excision to recover inoculated Salmonella and Campylobacter from broiler carcasses. In each of three replications, broiler carcass breast skin was inoculated with approximately 6.0 log antimicrobial-resistant Salmonella and Campylobacter, allowed to dry for 60 s, and sampled by either sponge, skin excision, or sponge followed by skin excision. Antimicrobial-resistant Salmonella and Campylobacter were enumerated from all samples. Skin excision allowed recovery of 0.1 to 0.2 log more inoculated bacteria than did sponge sampling. When excision was used on the same skin previously sampled by sponging, the combination of both methods did not significantly improve recovery compared with sponging alone. Skin excision is slightly more sensitive than sponge sampling; however, for repeated nondestructive sampling of broiler carcasses during processing, sponge sampling may be preferable to recover Salmonella and Campylobacter within 60 s of a contamination event.

  7. Oral inoculation of neonatal Suffolk sheep with the agent of classical scrapie results in PrP(Sc) accumulation in sheep with the PRNP ARQ/ARQ but not the ARQ/ARR genotype.

    Science.gov (United States)

    Greenlee, Justin J; Smith, Jodi D; Hamir, Amir N

    2016-04-01

    Scrapie is a transmissible spongiform encephalopathy that can be transmitted amongst susceptible sheep. The prion protein gene (PRNP) profoundly influences the susceptibility of sheep to the scrapie agent. This study reports the failure to detect PrP(Sc) in nervous or lymphoid tissues of Suffolk sheep of the PRNP ARQ/ARR genotype after oral inoculation with a U.S. scrapie isolate. Lambs were inoculated within the first 24 h of birth with 1 ml of a 10% (wt./vol.) brain homogenate derived from a clinically affected ARQ/ARQ sheep. The inoculated sheep were observed daily throughout the experiment for clinical signs suggestive of scrapie until they were necropsied at 86 months post inoculation. Tissues were collected for examination by immunohistochemistry and enzyme immunoassay, but all failed to demonstrate evidence of scrapie infection. Neonatal sheep of the ARQ/ARQ genotype receiving the same inoculum developed scrapie within 24 months. Lambs of the ARQ/ARR genotype that received the same inoculum by intracranial inoculation develop scrapie with a prolonged incubation period and with abnormal prion present within the central nervous system, but not peripheral lymphoid tissues. Results of this study suggest that ARQ/ARR sheep are resistant to oral infection with the scrapie isolate used even during the neonatal period.

  8. Network Inoculation: Heteroclinics and phase transitions in an epidemic model

    CERN Document Server

    Yang, Hui; Gross, Thilo

    2016-01-01

    In epidemiological modelling, dynamics on networks, and in particular adaptive and heterogeneous networks have recently received much interest. Here we present a detailed analysis of a previously proposed model that combines heterogeneity in the individuals with adaptive rewiring of the network structure in response to a disease. We show that in this model qualitative changes in the dynamics occur in two phase transitions. In a macroscopic description one of these corresponds to a local bifurcation whereas the other one corresponds to a non-local heteroclinic bifurcation. This model thus provides a rare example of a system where a phase transition is caused by a non-local bifurcation, while both micro- and macro-level dynamics are accessible to mathematical analysis. The bifurcation points mark the onset of a behaviour that we call network inoculation. In the respective parameter region exposure of the system to a pathogen will lead to an outbreak that collapses, but leaves the network in a configuration wher...

  9. Stress inoculation training: a case study in gymnastics.

    Science.gov (United States)

    Mace, R; Eastman, C; Carroll, D

    1986-09-01

    A young female gymnast of regional squad potential had ceased to make progress when she resumed training after a series of injuries and was given stress inoculation training to help her to regain her form. Preliminary interviews revealed that she had developed a number of negative self-statements and images which, it was hypothesised, may have been contributing towards her lack of progress. In order to replace these with positive self-statements and images a treatment programme of eight training sessions was implemented. Recorded interviews and subsequent comparison of comments made by the subject before and after the intervention programme, indicated that the training had been successful. This was endorsed by the coaches who reported an improved attitude to training and rapid progress in skill learning.

  10. INTRATHYMIC INOCULATION OF LIVER SPECIFIC ANTIGEN ALLEVIATES LIVER TRANSPLANT REJECTION

    Institute of Scientific and Technical Information of China (English)

    贾长库; 郑树森; 朱有法

    2004-01-01

    Objective To study the effects of liver specific antigen (LSA) on liver allotransplantation rejection. Methods Orthotopic liver transplantation was performed in this study. Group Ⅰ: syngeneic control (Wistar-to-Wistar); Group Ⅱ: acute rejection (SD-to-Wistar). Group Ⅲ: thymic inoculation of SD rat LSA day 7 before transplantation. The observation of general condition and survival time, rejection grades and the NF-κB activity of splenocytes were used to analyze severity of acute rejection and immune state of animals in different groups. Results The general condition of group Ⅰ was fair post transplantation with no sign of rejection. All recipients of group Ⅱ died within days 9 to 13 post transplantation with median survival time of 10.7 ±1.37 days. As for group Ⅲ, 5 out of 6 recipients survived for a long period with remarkably better general condition than that of group Ⅱ. Its rejection grades were significantly lower than group Ⅱ (P< 0.05).NF-κB activity was only detected in group Ⅰ between days 5 and 7 after transplantation, whereas high activity of NF-κB was detected at all points in group Ⅱ and low NF-κB activity was detected in group Ⅲ which was significantly lower than that of group Ⅱ (P < 0.05). Conclusions LSA is an important transplantation antigen directly involved in the immunorejection of liver transplantation. Intrathymic inoculation of LSA can alleviate the rejection of liver allotransplantation,grafts survive for a period of time thereby, allowing a novel way to liver transplantation immunotolerance.

  11. Ripening for improving the quality of inoculated cheese Rhizopus oryzae

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    ARTINI PANGASTUTI

    2010-01-01

    Full Text Available Estikomah SA, Sutarno, Pangastuti A 2010. Ripening for improving the quality of inoculated cheese Rhizopus oryzae. Nusantara Bioscience 2: 1-6. Cheese is dairy product resulted from fermented milk in which the fermentation process can be done by lactic acid bacteria or fungus. Rhizopus oryzae is able to produce lactic acid, protease and lipase. The ripening process changes the taste and texture. The purpose of this study is ripening to improve the quality of inoculated cheese R. oryzae. In this research the ripening was conducted the concentration variation of temperature (5oC, 10oC, 15oC, and time (7 days, 14 days. The procedure of research consisted of two steps, namely un-ripened cheese preparation followed by ripening cheese preparation. Cheese produced in this study analyzed the value of pH, fat content, protein content, amino acid levels and identification of microbe with ANOVA then followed by DMRT at 5% level of significance. Data results were analyzed with the like’s nonparametric statistical test, followed by Fridman Wilcoxon Signed Rank Test (WSRT at 5% level significance. The results showed that the preferred ripened cheese panelist was at a temperature of 15oC for 14 days. Ripening conditions affect pH, fat content, protein content and do not affect the levels of amino acids that formed ripened cheese. The best quality ripened cheese i.e. at a temperature of 15°C for 14 days, had a pH value of 4.40, the highest protein content of 9.78%, and fat content of 35.02%. The results of identified microbe in un-ripened cheese and ripened cheese include Enterococcus hirae (Enterococcus faecalis, Bacillus subtilis, and Aspergillus sp.

  12. The gamma interferon knockout mouse model for sarcocystis neurona: comparison of infectivity of sporocysts and merozoites and routes of inoculation.

    Science.gov (United States)

    Dubey, J P; Lindsay, D S; Kwok, O C; Shen, S K

    2001-10-01

    The dose-related infectivity of Sarcocystis neurona sporocysts and merozoites of 2 recent isolates of S. neurona was compared in gamma interferon knockout (KO) mice. Tenfold dilutions of sporocysts or merozoites were bioassayed in mice, cell culture, or both. All 8 mice, fed 1,000 sporocysts, developed neurological signs with demonstrable S. neurona in their tissues. Of 24 mice fed low numbers of sporocysts (100, 10, 1), 18 became ill by 4 wk postinoculation, and S. neurona was demonstrated in their brains; antibodies (S. neurona agglutination test) to S. neurona and S. neurona parasites were not found in tissues of the 6 mice that were fed sporocysts and survived for >39 days. One thousand culture-derived merozoites of these 2 isolates were pathogenic to all 8 mice inoculated subcutaneously (s.c.). Of the 24 mice inoculated s.c. with merozoites numbering 100, 10, or 1, only 3 mice had demonstrable S. neurona infection; antibodies to S. neurona were not found in the 21 mice that had no demonstrable organisms. As few as 10 merozoites were infective for cell cultures. These results demonstrate that at least 1,000 merozoites are needed to cause disease in KO mice. Sarcocystis neurona sporocysts were infective to mice by the s.c. route.

  13. Growth response of Pinus densiflora seedlings inoculated with three indigenous ectomycorrhizal fungi in combination

    Directory of Open Access Journals (Sweden)

    M Dalong

    2011-09-01

    Full Text Available Pinus densiflora seedlings were inoculated with three indigenous ectomycorrhizal fungi (Cenococcum geophilum, Rhizopogon roseolus and Russula densifolia in single-, two-, and three-species treatments. After 8 months, the colonization rates of each ectomycorrhizal species, seedling growth and the nutrition were assessed in each treatment. P. densiflora seedlings inoculated with different ECM species composition showed an increase in height and basal diameter and improved seedling root and shoot nutrition concentrations compared to control treatment. Generally, combined inoculation had a more positive influence on the seedlings than the single inoculation. The three-species inoculation presented the highest growth and basal diameter and concentration of most nutrients except potassium. In conclusion, the results provided strong evidence for benefits of combined inoculation with the indigenous ectomycorrhizal fungi on P. densiflora seedlings under controlled conditions.

  14. Growth response of Pinus densiflora seedlings inoculated with three indigenous ectomycorrhizal fungi in combination.

    Science.gov (United States)

    Dalong, M; Luhe, W; Guoting, Y; Liqiang, M; Chun, L

    2011-07-01

    Pinus densiflora seedlings were inoculated with three indigenous ectomycorrhizal fungi (Cenococcum geophilum, Rhizopogon roseolus and Russula densifolia) in single-, two-, and three-species treatments. After 8 months, the colonization rates of each ectomycorrhizal species, seedling growth and the nutrition were assessed in each treatment. P. densiflora seedlings inoculated with different ECM species composition showed an increase in height and basal diameter and improved seedling root and shoot nutrition concentrations compared to control treatment. Generally, combined inoculation had a more positive influence on the seedlings than the single inoculation. The three-species inoculation presented the highest growth and basal diameter and concentration of most nutrients except potassium. In conclusion, the results provided strong evidence for benefits of combined inoculation with the indigenous ectomycorrhizal fungi on P. densiflora seedlings under controlled conditions.

  15. Biochemical contents of pepper seedlings inoculated with phytophthora infestans and arbuscular mycorrhiza

    Directory of Open Access Journals (Sweden)

    Odebode A.C.

    2004-01-01

    Full Text Available The effect of interactions between Arbuscular Glomus etunicatum and fungus Phytophthora infestans on biochemical contents of pepper plants was investigated in a greenhouse experiment. The sugar contents (i.e. Glucose fructose and sucrose were higher in the control and mycorrhizal inoculated pepper seedlings and the lowest in pathogen inoculated seedlings. Free amino acids were the highest in the simultaneously inoculated pepper seedlings while total phenol was found to be the highest in pepper seedlings inoculated with P. infestans. The levels of nitrogen, phosphorus and potassium varied in the inoculated pepper seedlings without any significant difference in the treatment. The results obtained suggest protective influence of mycorrhiza by enhancing the nutritional status of the inoculated pepper seedlings.

  16. Paddy plants inoculated with PGPR show better growth physiology and nutrient content under saline condition

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    Yachana Jha

    2013-09-01

    Full Text Available The possible role of plant growth-promoting rhizobacteria (PGPR to alleviate salt stress during plant growth has been studied on paddy rice (Oryza sativa L. 'GJ-17' under greenhouse conditions; the study included growth parameters, mineral concentration, and antioxidant enzyme level. Salinity reduced plant growth, but PGPR inoculation reduced its harmful effect up to 1% salinity. Plants inoculated with PGPR under saline conditions showed 16% higher germination, 8% higher survival, 27% higher dry weight, and 31% higher plant height. Similarly, PGPR inoculated plants showed increased concentrations of N(26%, P (16%, K (31%, and reduced concentrations of Na (71% and Ca (36% as compared to non-inoculated control plants under saline conditions. Plants inoculated with PGPR under saline conditions also showed significant variations in antioxidant levels and growth physiology. Results suggested that inoculation with PGPR Bacillus pumilus and Pseudomonas pseudoalcaligenes in salt-stressed plants could help to alleviate salt stress in the paddy.

  17. The roles of inoculants' carbon source use in the biocontrol of potato scab disease.

    Science.gov (United States)

    Sun, Pingping; Zhao, Xinbei; Shangguan, Nini; Chang, Dongwei; Ma, Qing

    2015-04-01

    Despite the application of multiple strains in the biocontrol of plant diseases, multistrain inoculation is still constrained by its inconsistency in the field. Nutrients, especially carbons, play an important role in the biocontrol processes. However, little work has been done on the systematic estimation of inoculants' carbon source use on biocontrol efficacies in vivo. In the present study, 7 nonpathogenic Streptomyces strains alone and in different combinations were inoculated as biocontrol agents against the potato scab disease, under field conditions and greenhouse treatments. The influence of the inoculants' carbon source use properties on biocontrol efficacies was investigated. The results showed that increasing the number of inoculated strains did not necessarily result in greater biocontrol efficacy in vivo. However, single strains with higher growth rates or multiple strains with less carbon source competition had positive effects on the biocontrol efficacies. These findings may shed light on optimizing the consistent biocontrol of plant disease with the consideration of inoculants' carbon source use properties.

  18. Degradation of Dissolved Organic Carbon from Discontinuous Permafrost Due to Photolysis and Different Inoculants

    Science.gov (United States)

    Aukes, P.; Schiff, S. L.

    2013-12-01

    Northern areas with permafrost are very susceptible to a warming climate. Temperature increases can alter hydrologic flow paths, increase the depth and biogeochemistry of the active layer, and degrade and reduce the amount of remaining permafrost. Particularly, loss of permafrost will release large stores of previously unavailable frozen carbon to the environment. Dissolved organic carbon (DOC) plays many important roles that affect both ecosystem health and drinking water quality. Comprised of countless different molecules, DOC absorbs harmful ultra-violet (UV) radiation and controls thermal regimes of lakes, is an important energy and nutrient source for heterotrophic microbes, complexes with and transports heavy metals, and reacts during chlorination of drinking water to form carcinogenic disinfection by-products. Since the ultimate fate of DOC depends on its reactivity with the surrounding environment, the implications of DOC released from permafrost for ecosystems and drinking water quality will vary across the landscape. We used 90-day lab incubations to assess the differences in quality of DOC by observing the susceptibility for DOC to degrade among various discontinuous-permafrost sources. Specifically, UV-photolysis and two surface water inoculants (pond and creek water filtered to 2.0μm) were used to represent the dominant degradation pathways encountered within the environment. Samples were taken in July 2013 from three locations (pond, creek, and wetland porewater) in a region of discontinuous permafrost near Yellowknife, NWT, Canada. We observed changes to the composition and quality of DOC resulting from photolysis and degradation by two inoculants over 90 days, where DOC quality was determined by Liquid Chromatography - Organic Carbon Detection, DOC:DON, UV-absorbance, and changes to other constituents (DIC, δ13C-DIC, CO2). We hypothesize that UV-photolysis and microbial degradation will readily degrade easily accessible and reactive components of

  19. Co-inoculation of Azospirillum brasilense and Herbaspirillum seropedicae in maize

    OpenAIRE

    Janaína Dartora; Deniele Marini; Edilaine. D. V. Gonçalves; Vandeir F. Guimarães

    2016-01-01

    ABSTRACT Bacteria from the genera Azospirillum and Herbaspirillum have been associated with increments in maize yield. The aim of this study was to evaluate the maize yield and nutritional content in response to inoculation with A. brasilense and H. seropediceae in association with nitrogen (N) fertilization. The experimental design was randomized blocks in a 4 x 5 factorial scheme, with four replicates. The treatments consisted of seed inoculation (control without N and inoculation, A. brasi...

  20. Seed quality, chlorophyll content index and leaf nitrogen levels in maize inoculated with Azospirillum brasilense

    OpenAIRE

    Lívia de Matos Pereira; Elise de Matos Pereira; Lucas Tadeu Mazza Revolti; Sonia Marli Zingaretti; Gustavo Vitti Môro

    2015-01-01

    The aim of this study was to evaluate differences between maize genotypes in relation to the germination response of the seeds and the growth of seedlings inoculated with Azospirillum brasilense, as well as the effect of inoculation on nitrogen levels and the chlorophyll content index of the leaves. The physiological seeds quality from the single-cross hybrids AG7098 and 2B707, and from the experimental synthetic varieties V2 and V4, inoculated with A. brasilense, was tested for germination, ...

  1. Analysis of 30 patients with acupuncture-induced primary inoculation tuberculosis.

    Science.gov (United States)

    Liu, Yangbo; Pan, Jingye; Jin, Keke; Liu, Cailong; Wang, Jing; Chen, Li; Chen, Lei; Yuan, Jiandong

    2014-01-01

    Primary inoculation tuberculosis is a skin condition that develops at the site of inoculation of Mycobacterium tuberculosis in tuberculosis-free individuals. This report describes the diagnosis, treatment and >1 year follow-up of 30 patients presenting with acupuncture-induced primary inoculation tuberculosis. Our data provide a deeper insight into this rare route of infection of tuberculosis. We also review effective treatment options.

  2. FERMENTATION QUALITY OF KING GRASS SILAGE TREATED WITH LIQUID OR DRIED INOCULANT OF LACTIC ACID BACTERIA

    Directory of Open Access Journals (Sweden)

    B. Santoso

    2016-03-01

    Full Text Available The aim of this study was to evaluate the nutritive value and fermentation characteristic of king grass silage treated with addition of liquid or dried lactic acid bacteria (LAB inoculant. Experiment was arranged to a completely randomized design with four treatments and three replications. Four treatments as follows (A king grass without LAB inoculant as the control; (B king grass + 30 ml of liquid of epiphytic LAB inoculant/kg of fresh forage; (C king grass + 30 g of freeze-dried powder of LAB inoculant/kg of fresh forage; (D king grass + 30 g of centrifuged powder of LAB inoculant/kg of fresh forage. The liquid LAB inoculant was sprayed on the top of grass and then mannualy mixed by hand. The intial LAB concentration in all inoculants were 1.0 × 106 cfu/g. About 500 g of silage materials were ensiled into plastic silos and stored at room temperatures (approximately 28oC for 30 days. Results showed that pH value, concentrations of lactic acid, N-amonia (NH3-N, butyric acid and total volatile fatty acids (VFA as well as Fleigh Point were affected by treatment of LAB inoculant. Silage treated with liquid or dried of LAB inoculant had lower (P<0.01 pH value compared to the control silage. Concentrations of NH3-N and butyric acid significantly decreased (P<0.01 in silage treated with LAB inoculants. Silage treated with centrifuged powder of LAB inoculant had the highest (P<0.05 Fleigh Point than other silage. In vitro neutral detergent fiber (NDF digestibility was significantly higher (P<0.05 in silage treated with LAB inoculant compared to the control silage.

  3. Co-inoculation of Azospirillum brasilense and Herbaspirillum seropedicae in maize

    OpenAIRE

    Janaína Dartora; Deniele Marini; Edilaine. D. V. Gonçalves; Guimarães, Vandeir F. [UNESP

    2016-01-01

    ABSTRACT Bacteria from the genera Azospirillum and Herbaspirillum have been associated with increments in maize yield. The aim of this study was to evaluate the maize yield and nutritional content in response to inoculation with A. brasilense and H. seropediceae in association with nitrogen (N) fertilization. The experimental design was randomized blocks in a 4 x 5 factorial scheme, with four replicates. The treatments consisted of seed inoculation (control without N and inoculation, A. brasi...

  4. Characteristics of filiform, fungiform and vallate papillae and surface of interface epithelium-connective tissue of the maned sloth tongue mucosa (Bradypus torquatus, Iliger, 1811): Light and Scanning Electron Microscopy Study.

    Science.gov (United States)

    Benetti, E J; Pícoli, L C; Guimarães, J P; Motoyama, A A; Miglino, M A; Watanabe, L-S

    2009-02-01

    The study of lingual surfaces and the surface of interface epithelium-connective tissue of the tongue of Bradypus torquatus was performed by employing the light and scanning electron microscopy (SEM) techniques. The results revealed that the rostral part of the tongue presents a round apex and covered by filiform and fungiform lingual papillae and a ventral smooth surface. It was observed that the epithelial layer of the dorsal surface possesses the basal, spinosum, granular and cornified epithelial cells. The lamina propria is characterized by a dense connective tissue forming the long, short and round papillae. Numerous typical filiform papillae are located especially in the rostral part intermingled for few fungiform papillae, which were revealed in three-dimensional SEM images. Usually, the fungiform papillae are located in the border of rostral apex of the tongue exhibiting the rounded form. They are covered by keratinized epithelial cells. In the fungiform papillae, several taste pores were observed on the surface. The vallate papillae presented numerous taste buds in the wall of epithelial cells, being that the major number of taste buds is located on the superior half of vallate papilla. The taste pores are surrounded by several laminae of keratinized epithelial cells. The samples treated with NaOH solution and examined by SEM revealed, after removal of the epithelial layer, the dense connective core in original disposition, presenting different sizes and shapes. The specimens stained with Picrosirius and examined by polarized light microscopy revealed the connective tissue, indicating the collagen fibres type I and type III.

  5. Inoculated fermentation of green olives with potential probiotic Lactobacillus pentosus and Lactobacillus plantarum starter cultures isolated from industrially fermented olives.

    Science.gov (United States)

    Blana, Vasiliki A; Grounta, Athena; Tassou, Chrysoula C; Nychas, George-John E; Panagou, Efstathios Z

    2014-04-01

    The performance of two strains of lactic acid bacteria (LAB), namely Lactobacillus pentosus B281 and Lactobacillus plantarum B282, previously isolated from industrially fermented table olives and screened in vitro for probiotic potential, was investigated as starter cultures in Spanish style fermentation of cv. Halkidiki green olives. Fermentation was undertaken at room temperature in two different initial salt concentrations (8% and 10%, w/v, NaCl) in the brines. The strains were inoculated as single and combined cultures and the dynamics of their population on the surface of olives was monitored for a period of 114 days. The survival of inoculated strains on olives was determined using Pulsed Field Gel Electrophoresis (PFGE). Both probiotic strains successfully colonized the olive surface at populations ranged from 6.0 to 7.0 log CFU/g throughout fermentation. PFGE analysis revealed that L. pentosus B281 presented higher colonization in both salt levels at the end of fermentation (81.2% and 93.3% in 8% and 10% NaCl brines, respectively). For L. plantarum B282 a high survival rate (83.3%) was observed in 8% NaCl brines, but in 10% NaCl the strain could not colonize the surface of olives. L. pentosus B281 also dominated over L. plantarum B282 in inoculated fermentations when the two strains were used as combined culture. The biochemical profile (pH, organic acids, volatile compounds) attained during fermentation and the sensory analysis of the final product indicated a typical lactic acid fermentation process of green olives.

  6. Cholesterol enrichment of human monocyte/macrophages induces surface exposure of phosphatidylserine and the release of biologically-active tissue factor-positive microvesicles.

    Science.gov (United States)

    Liu, Ming-Lin; Reilly, Michael P; Casasanto, Peter; McKenzie, Steven E; Williams, Kevin Jon

    2007-02-01

    Biologically significant amounts of two procoagulant molecules, phosphatidylserine (PS) and tissue factor (TF), are transported by monocyte/macrophage-derived microvesicles (MVs). Because cellular cholesterol accumulation is an important feature of atherosclerotic vascular disease, we now examined effects of cholesterol enrichment on MV release from human monocytes and macrophages. Cholesterol enrichment of human THP-1 monocytes, alone or in combination with lipopolysaccharide (LPS), tripled their total MV generation, as quantified by flow cytometry based on particle size and PS exposure. The subset of these MVs that were also TF-positive was likewise increased by cellular cholesterol enrichment, and these TF-positive MVs exhibited a striking 10-fold increase in procoagulant activity. Moreover, cholesterol enrichment of primary human monocyte-derived macrophages also increased their total as well as TF-positive MV release, and these TF-positive MVs exhibited a similar 10-fold increase in procoagulant activity. To explore the mechanisms of enhanced MV release, we found that cholesterol enrichment of monocytes caused PS exposure on the cell surface by as early as 2 hours and genomic DNA fragmentation in a minority of cells by 20 hours. Addition of a caspase inhibitor at the beginning of these incubations blunted both cholesterol-induced apoptosis and MV release. Cholesterol enrichment of human monocyte/macrophages induces the generation of highly biologically active, PS-positive MVs, at least in part through induction of apoptosis. Cholesterol-induced monocyte/macrophage MVs, both TF-positive and TF-negative, may be novel contributors to atherothrombosis.

  7. Carbon decomposition by inoculating Phanerochaete chrysosporium during drum composting of agricultural waste.

    Science.gov (United States)

    Varma, V Sudharsan; Ramu, Kamma; Kalamdhad, Ajay S

    2015-05-01

    The effect of Phanerochaete chrysosporium inoculation during drum composting of agricultural waste was performed at different composting stages. Three trials were carried out with (5:4:1) combination of vegetable waste, cattle manure, and sawdust along with 10 kg of dried leaves with a total mass of 100 kg in a 550 L rotary drum composter. Trial 1 was a control without inoculation of fungus, while trial 2 was inoculated during the initial day (0th day of composting), and trial 3 was inoculated after the thermophilic phase, i.e., on the 8th day of composting period. The inoculation of fungus increased the volatile solids reduction by 1.45-fold in trial 2 and 1.7-fold in trial 3 as compared to trial 1 without any fungal inoculation. Total Kjeldahl Nitrogen (TKN) was observed with 2.31, 2.62, and 2.59% in trials 1, 2, and 3, respectively, at the end of 20 days of composting period. Hence, it can be concluded that inoculation of white-rot fungus increased the decomposition rate of agricultural waste within shorter time in drum composting. However, inoculation after the thermophilic phase was found more effective than inoculation during initial days of composting for producing more stabilized and nutrient-rich compost.

  8. Establishment of Desmoncus orthacanthos Martius (Arecaceae): effect of inoculation with arbuscular mycorrhizae.

    Science.gov (United States)

    Ramos-Zapata, José A; Orellana, Roger; Allen, Edith B

    2006-03-01

    Inoculation with arbuscular mycorrhizal (AM) fungi has often promoted increased growth of plants but very little work has been done in the tropics to evaluate the effects of inoculation on the establishment and development of seedlings in forests. Desmoncus orthacanthos Martius is a scandent palm present both in early and late succession, and consequently can be used in restoration processes. A test was conducted to determine the effect of AM on the establishment of Desmoncus orthacanthos in tropical forest in the Yucatan Peninsula, Mexico. Thirty inoculated and 30 non-inoculated seedlings were introduced in two sites of different successional age, a mature forest and an eight-year old abandoned cornfield (acahual). Survival and growth parameters were evaluated after 12 months. Leaf area and phosphorus, but not height, were greater in inoculated than non-inoculated plants in the forest but not in the acahual. However, mycorrhizae had a clear effect on plant survival in both sites, with a threefold increase in survival of inoculated compared with non-inoculated plants bassed on an odds ratio. The results suggest that inoculation will be important to increase the establishment of this commercially important palm.

  9. Effects of inoculation with arbuscular mycorrhizal fungi on maize grown in multi-metal contaminated soils.

    Science.gov (United States)

    Liang, Chang-Cong; Li, Tao; Xiao, Yan-ping; Liu, Mao-Jun; Zhang, Han-Bo; Zhao, Zhi-Wei

    2009-01-01

    Pot culture experiments were established to determine the effects of colonization by arbuscular mycorrhizal fungi (AMF) (Glomus mosseae and G. sp) on maize (Zea mays L.) grown in Pb, Zn, and Cd complex contaminated soils. AMF and non-AMF inoculated maize were grown in sterilized substrates and subjected to different soil heavy metal (Pb, Zn, Cd) concentrations. The root and shoot biomasses of inoculated maize were significantly higher than those of non-inoculated maize. Pb, Zn, and Cd concentrations in roots were significantly higher than those in shoots in both the inoculated and non-inoculated maize, indicating the heavy metals mostly accumulated in the roots of maize. The translocation rates of Pb, Zn, and Cd from roots to shoots were not significantly difference between inoculated and non-inoculated maize. However, at high soil heavy metal concentrations, Pb, Zn, and Cd in the shoots and Pb in the roots of inoculated maize were significantly reduced by about 50% compared to the non-inoculated maize. These results indicated that AMF could promote maize growth and decrease the uptake of these heavy metals at higher soil concentrations, thus protecting their hosts from the toxicity of heavy metals in Pb, Zn, and Cd complex contaminated soils.

  10. Histopathological study of response of Vitis vinifera cv. Cabernet Sauvignon to bark and wood injury with and without inoculation by Phaeomoniella chlamydospora

    Directory of Open Access Journals (Sweden)

    Jérôme POUZOULET

    2013-09-01

    Full Text Available Summary. Phaeomoniella chlamydospora (Pch is one of the main causal agents of tracheomycosis in grapevine. We characterize how this fungus affects the response of Vitis vinifera cv. Cabernet Sauvignon to bark and xylem-tissue wounding after six weeks post-treatment. A histological investigation shows that, in xylem tissue, cell-wall modifications in response to wounding are related to suberin deposits rather than to lignin-induced wall thickening. The xylem response does not appear to be disturbed by Pch infection. Therefore, cell-wall modification strongly inhibits the development of wound-closure tissue (WCT but does not prevent the differentiation of the necro-phylactic periderm. Hyphae localization in tissue surrounding the wound or inoculation sites indicates that Pch colonizes all cell types, such as vascular tissues, paratracheal parenchyma cells, fibers and rays. The results also suggest that efficient compartmentalization separating fascicular xylem portions is assured by thick suberized cell walls bordering the ray parenchyma.

  11. Influence of the season in the development of symptoms in plants of ‘Grande naine’ artificially inoculated with Mycosphaerella fijiensis in greenhouse

    Directory of Open Access Journals (Sweden)

    Michel Leiva-Mora

    2011-01-01

    Full Text Available Plant pathogen fungi required favorable conditions to invade and colonize plant tissues, which may change according to the season of the year. This work is aimed to evaluate the influence of different seasons (June-August and August-October in the development of symptoms in ‘Grande naine’ artificially inoculated with Mycosphaerella fijiensis in greenhouse. Inoculation assays were done during three consecutive years (2005, 2006 and 2007. None differences in attack intensity between different seasons were observed, at 14, 21, 35, 49 y 63 days post inoculation (dpi. This shows that the development of symptoms in greenhouse was similar in the period from June to October. Temperature and humidity have not been controlled by all laboratories that conduct studies related to M. fijiensis. Then, to choose the appropriate season for artificial inoculations is a prerequisite for effective development of M. fijiensis infective cycle and to avoid masking in the phenotype of resistance. Key words: attack intensity, Black Sigatoka, mycelia

  12. Solubilisation of inorganic phosphates by inoculant strains from tropical legumes

    Directory of Open Access Journals (Sweden)

    Leandro Marciano Marra

    2011-10-01

    Full Text Available Microbial solubilisation of low soluble inorganic phosphates is an important process contributing for the phosphorus available to plants in tropical soils. This study evaluates the ability of inoculant strains for tropical legumes to solubilise inorganic phosphates of low solubility that are found in tropical soils. Seven strains of Leguminosae nodulating bacteria (LNB were compared with one another and with a non-nodulating positive control, Burkholderia cepacia (LMG 1222T. Four of the strains are used as inoculants for cowpeas (Vigna unguiculata (Bradyrhizobium sp. UFLA 03-84; Bradyrhizobium elkani INPA 03-11B and Bradyrhizobium japonicum BR3267 or for common beans (Phaseolus vulgaris (Rhizobium tropici CIAT 899T. Rhizobium etli UFLA 02-100 and Rhizobium leguminosarum 316C10a are also efficient nodulators of beans and Cupriavidus taiwanensis LMG 19424T nodulates on Mimosa pudica. Two experiments, with solid and liquid media, were performed to determine whether the strains were able to solubilise CaHPO4, Al(H2PO43 or FePO4.2H2O. On solid GELP medium none of the strains dissolved FePO4.2H2O, but LMG 1222, UFLA 03-84 and CIAT 899 solubilised CaHPO4 particularly well. These strains, along with LMG 19424 and BR 3267, were also able to increase the solubility of Al(H2PO43. In liquid GELP medium, LMG 1222 solubilised all phosphate sources, but no legume nodulating strain could increase the solubility of Al(H2PO43. The strains CIAT 899 and UFLA 02-100 were the only legume nodulating bacteria able to solubilise the other phosphate sources in liquid media, dissolving both CaHPO4 and FePO4.2H2O. There was a negative correlation between the pH of the culture medium and the concentration of soluble phosphate when the phosphorus source was CaHPO4 or FePO4.2H2O. The contribution of these strains to increasing the phosphorus nutrition of legumes and non-legume plant species should be investigated further by in vivo experiments.

  13. Quality assessment of truffle-inoculated seedlings in Italy: proposing revised parameters for certification

    Directory of Open Access Journals (Sweden)

    Domizia Donnini

    2014-08-01

    Full Text Available Aim of study: the main aims of this study were to evaluate the quality of truffle-inoculated seedlings produced by commercial nurseries in Italy and to identify their minimum requisites in terms of plant age, health, homogeneity, and cut-off percentage of inoculated Tuber and non-Tuber ectomycorrhizae, based on the analysis of an extensive sample of seedlings subjected to quality control and certification.Area of study: truffle-inoculated seedlings produced by Italian commercial nurseries.Material and Methods: analysis of truffle-inoculated seedlings for health and quality standards; recording of presence of inoculated Tuber spp. and other concurrent fungi according to the official Italian method for certification; selective amplification of ectomycorrhizal DNA by PCR species-specific primers.Main results: We showed that mycorrhization levels in truffle-inoculated seedlings increased with time after truffle-spore inoculation. The highest mean percentage of the inoculated Tuber spp., but also the highest presence of contaminants, were recorded after three years. The mycorrhization level of Tuber melanosporum and T. aestivum was higher in Corylus and Ostrya seedlings than in Q. ilex and Q. pubescens, but the latter two host species showed the lowest presence of other ectomycorrhizal fungi. Mycorrhization level distribution in truffle-inoculated seedlings of suitable batches differed very little from the distribution in only all suitable seedlings. Truffle seedlings with other Tuber spp. were very few and even absent after three years. The general quality of Italian truffle-inoculated seedlings is high but can be improved even further by revising the parameters used for their certification.Research highlights: Mycorrhization assessment in truffle-inoculated seedlings produced by commercial nurseries and a revision of the parameters of quality standards following several years of certification in Italy.Keywords: Truffle cultivation; truffle

  14. Soil inoculation with symbiotic microorganisms promotes plant growth and nutrient transporter genes expression in durum wheat.

    Science.gov (United States)

    Saia, Sergio; Rappa, Vito; Ruisi, Paolo; Abenavoli, Maria Rosa; Sunseri, Francesco; Giambalvo, Dario; Frenda, Alfonso S; Martinelli, Federico

    2015-01-01

    In a field experiment conducted in a Mediterranean area of inner Sicily, durum wheat was inoculated with plant growth-promoting rhizobacteria (PGPR), with arbuscular mycorrhizal fungi (AMF), or with both to evaluate their effects on nutrient uptake, plant growth, and the expression of key transporter genes involved in nitrogen (N) and phosphorus (P) uptake. These biotic associations were studied under either low N availability (unfertilized plots) and supplying the soil with an easily mineralizable organic fertilizer. Regardless of N fertilization, at the tillering stage, inoculation with AMF alone or in combination with PGPR increased the aboveground biomass yield compared to the uninoculated control. Inoculation with PGPR enhanced the aboveground biomass yield compared to the control, but only when N fertilizer was added. At the heading stage, inoculation with all microorganisms increased the aboveground biomass and N. Inoculation with PGPR and AMF+PGPR resulted in significantly higher aboveground P compared to the control and inoculation with AMF only when organic N was applied. The role of microbe inoculation in N uptake was elucidated by the expression of nitrate transporter genes. NRT1.1, NRT2, and NAR2.2 were significantly upregulated by inoculation with AMF and AMF+PGPR in the absence of organic N. A significant down-regulation of the same genes was observed when organic N was added. The ammonium (NH4 (+)) transporter genes AMT1.2 showed an expression pattern similar to that of the NO3 (-) transporters. Finally, in the absence of organic N, the transcript abundance of P transporters Pht1 and PT2-1 was increased by inoculation with AMF+PGPR, and inoculation with AMF upregulated Pht2 compared to the uninoculated control. These results indicate the soil inoculation with AMF and PGPR (alone or in combination) as a valuable option for farmers to improve yield, nutrient uptake, and the sustainability of the agro-ecosystem.

  15. The Effect of Microbial Inoculants Applied at Ensiling on Sorghum Silage Characteristics and Aerobic Stability

    Institute of Scientific and Technical Information of China (English)

    GUAN Wu-tai; Ashbell G; Hen Y; Weinberg Z G

    2002-01-01

    Whole crop forage sorghum (Saccharatum) cultivar FS5 was harvested at the soft dough stage of maturity. The sorghum was chopped to approximately 2 cm pieces and ensiled under laboratory conditions in 1.5 L Weck glass jars. At ensiling, it was treated with two commercial microbial inoculants: inoculant A and inoculant B. The inoculants were applied at 2 × 105 colony forming units g- 1 DM. Silage with no additives served as a control. Three jars per treatment were opened on days 2, 4, 8, 15 and 60 post-ensiling to study fermentation dynamics. After 60 days of ensiling, the silages were analyzed and subjected to an aerobic stability test lasting 5 days. Results showed that both inoculants caused a more rapid rate of pH decline and a higher amount of lactic acid production. Silages treated with each inoculant produced a little more CO2 and resulted in more glucose loss as compared with the control. Addition of inoculants did not influence ( P > 0.05) the ash and crude protein contents, but tended to decrease the concentration of acetic acid (P < 0.05), butyric acid (P < 0.01) and propionic acid ( P < 0.01 ), and increase the lactic acid concentration ( P < 0.01 ). Silages treated with inoculant A possess the more DM loss, and the higher yeast counts upon aerobic exposure. Silage treated with inoculant B had the most DM (P < 0.05), lactic acid contents (P < 0.01 ), the least acetic acid content (P < 0.05). Inoculant B reduced the ADF (P < 0.01), ADL and NDF ( P < 0.05) contents. It was concluded that lactic bacteria inoculants may improve the fermentation but might impair the aerobic stability for sorghum ensilage.

  16. Effects of Lactic Acid Bacteria Inoculated Fermentation on Pickled Cucumbers

    Directory of Open Access Journals (Sweden)

    Xiaoyi Ji

    2013-12-01

    Full Text Available The aim of this study was to determine the effects of Lactic Acid Bacteria (LAB fermentation on the texture and organic acid of pickled cucumbers. Texture and sensory evaluation as well as a microscopic observation were performed to study the textural differences among fresh cucumber, Spontaneous fermentation (SF cucumber and LAB Inoculating Fermentation (LABIF cucumber. Accumulation of seven organic acids i.e., oxalic, tartaric, malic, lactic, acetic, citric and succinic acid during cucumber pickling were also studied. The disruption extent of the middle lamella in SF cucumber displayed more obviously than that in LABIF cucumber, implying that LABIF contributed to keep the cucumber original structure intact. Based on the organic acid accumulation pattern, in SF LAB and Acetic Acid Bacteria (AAB fermented simultaneously, while in LABIF LAB fermented beforehand thus being in dominant position, then AAB fermented vigorously in a acidic condition created by LAB. The acetic acid accumulaton pattern could be regarded as the distinctive feature between SF and LABIF. The orgnic acids produced in LABIF were higher than that in SF. The final score of sensory evaluation combining texture analysis demonstrated that LABIF overmatched SF. It was concluded that LABIF could obviously enhance the quality of pickled cucumber and overwhelming SF, due to LABIF more beneficial to keep the cucumber original structure intact and organic acids accumulation.

  17. Microbial inoculants and fertilization for bioremediation of oil in wetlands

    Energy Technology Data Exchange (ETDEWEB)

    Neralla, S.; Wright, A.L.; Weaver, R.W. [Texas A and M Univ., College Station, TX (United States). Dept. of Soil and Crop Sciences

    1995-12-31

    Bioremediation is an attractive alternative to physical methods of oil spill cleanup in wetlands where the ecosystem can be easily damaged. Because populations of oil-degrading microorganisms are usually low in wetlands, there is potential for increasing bioremediation through bioaugmentation in conjunction with N and P supplementation. Eight microbial inoculant products were added to microcosms containing soil from a salt marsh. Four of these products were also used in mesocosms containing Spartina alterniflora grown in a glasshouse. In unfertilized microcosms, the extent of oil degraded as measured by carbon dioxide evolution during 90 days, was 30% higher in the product with the highest activity than was recorded in the control with oil by 36%. None of the products when added to the fertilized soil increased activity above that of the fertilized control with oil. Addition of oil to microcosms increased populations of hydrocarbon-degrading microorganisms, but bioaugmentation products did not increase populations. Neither addition of products nor fertilization enhanced the disappearance of oil in mesocosms in the glasshouse. Approximately 50% of the weathered oil disappeared in 41 d for all treatments. Because bioaugmentation did not enhance oil degradation, it seems that natural populations of hydrocarbon-degrading microorganisms were adequate in the salt marsh soil for bioremediation.

  18. Uropathogenic Escherichia coli induces serum amyloid a in mice following urinary tract and systemic inoculation.

    Directory of Open Access Journals (Sweden)

    Andreja Erman

    Full Text Available Serum amyloid A (SAA is an acute phase protein involved in the homeostasis of inflammatory responses and appears to be a vital host defense component with protective anti-infective properties. SAA expression remains poorly defined in many tissues, including the urinary tract which often faces bacterial challenge. Urinary tract infections (UTIs are usually caused by strains of uropathogenic Escherichia coli (UPEC and frequently occur among otherwise healthy individuals, many of whom experience bouts of recurrent and relapsing infections despite the use of antibiotics. To date, whether SAA is present in the infected urothelium and whether or not the induction of SAA can protect the host against UPEC is unclear. Here we show, using mouse models coupled with immunofluorescence microscopy and quantitative RT-PCR, that delivery of UPEC either directly into the urinary tract via catheterization or systemically via intraperitoneal injection triggers the expression of SAA. As measured by ELISA, serum levels of SAA1/2 were also transiently elevated in response to UTI, but circulating SAA3 levels were only up-regulated substantially following intraperitoneal inoculation of UPEC. In in vitro assays, physiological relevant levels of SAA1/2 did not affect the growth or viability of UPEC, but were able to block biofilm formation by the uropathogens. We suggest that SAA functions as a critical host defense against UTIs, preventing the formation of biofilms both upon and within the urothelium and possibly providing clinicians with a sensitive serological marker for UTI.

  19. Effect of Bacterial—Feeding Uematode Inoculation on Wheat Growth and N and P Uptake

    Institute of Scientific and Technical Information of China (English)

    LIHULXIN; HUFENG

    2001-01-01

    A 40-day gnotobiotic microcosm experiment was carried out to quantify the effect of bacterial-feeding nematode on plat growth and nutrient absorption.The results showed that inoculation of bacterial-feeding nematode Protorhabditis sp.stimulated the growth of wheat (Triticum aestivum) and the uptake of N.By the end of the 40-day incubation wheat biomass and N uptake in the treatment with nematode and bacteria (Pseudomonas sp.)increased by 6.5% and 5.9%,respectively,compared with bacteria alone treatment.The presence of nematode mainly accelerated the growth of aboveground of wheat,while it slightly inhibited the root development.There was little difference in plant tissue N concentration between treatments.P concentration and uptake of wheat,however,were generally reduced by nematode, It appears that the enhancement of plant growth and nitrogen uptake is attributed to the enhancement of nitrogen mineraliztion induced by nematode feeding on bacteria,and the reduction of phosphorous uptake is the result of ewak root status and comptetition by bacteria immobilzation.

  20. Stromata, sporangiomata and chlamydosori of Phytophthora ramorum on inoculated Mediterranean woody plants.

    Science.gov (United States)

    Moralejo, Eduardo; Puig, Miquel; García, José A; Descals, Enrique

    2006-11-01

    Three types of multihyphal structures, stromata, sporangiomata and chlamydosori, are described for the plant pathogen Phytophthora ramorum. Their morphology, morphogenesis and position on the host organ were observed by dissecting, compound and scanning electron microscopy. Stromata were consistently formed one to two weeks after zoospore inoculation of detached leaves and fruits of an assortment of Mediterranean sclerophyll shrubs. Stroma initials appeared subcuticularly or subepidermally and developed as small hyphal aggregates by repeated branching, budding, swelling and interweaving, eventually forming a prosenchyma. They always emerged through the adaxial side of the leaf by rupture of the overlying host tissue. Occasionally sporangia and chlamydosori (packed clusters of chlamydospores) were formed on the stromata. Sporangiomata bore short sporangiophores and clusters of 20-100 sporangia and resembled sporodochia of the mitosporic fungi. The biological significance of these multihyphal structures is discussed. Some epidemiological aspects were also studied: several understorey species of the holm oak (Quercus ilex) woodland were susceptible to in vitro infection with three isolates of P. ramorum originally collected from different ornamental hosts. The risk of spread to this ecosystem is evaluated.

  1. Root inoculation with Pseudomonas putida KT2440 induces transcriptional and metabolic changes and systemic resistance in maize plants.

    Science.gov (United States)

    Planchamp, Chantal; Glauser, Gaetan; Mauch-Mani, Brigitte

    2014-01-01

    Pseudomonas putida KT2440 (KT2440) rhizobacteria colonize a wide range of plants. They have been extensively studied for their capacity to adhere to maize seeds, to tolerate toxic secondary metabolites produced by maize roots and to be attracted by maize roots. However, the response of maize plants to KT2440 colonization has not been investigated yet. Maize roots were inoculated with KT2440 and the local (roots) and systemic (leaves) early plant responses were investigated. The colonization behavior of KT2440 following application to maize seedlings was investigated and transcriptional analysis of stress- and defense-related genes as well as metabolite profiling of local and systemic maize tissues of KT2440-inoculated were performed. The local and systemic responses differed and more pronounced changes were observed in roots compared to leaves. Early in the interaction roots responded via jasmonic acid- and abscisic acid-dependent signaling. Interestingly, during later steps, the salicylic acid pathway was suppressed. Metabolite profiling revealed the importance of plant phospholipids in KT2440-maize interactions. An additional important maize secondary metabolite, a form of benzoxazinone, was also found to be differently abundant in roots 3 days after KT2440 inoculation. However, the transcriptional and metabolic changes observed in bacterized plants early during the interaction were minor and became even less pronounced with time, indicating an accommodation state of the plant to the presence of KT2440. Since the maize plants reacted to the presence of KT2440 in the rhizosphere, we also investigated the ability of these bacteria to trigger induced systemic resistance (ISR) against the maize anthracnose fungus Colletotrichum graminicola. The observed resistance was expressed as strongly reduced leaf necrosis and fungal growth in infected bacterized plants compared to non-bacterized controls, showing the potential of KT2440 to act as resistance inducers.

  2. Allergic airway inflammation by nasal inoculation of particulate matter (PM2.5 in NC/Nga mice.

    Directory of Open Access Journals (Sweden)

    Keiki Ogino

    Full Text Available To evaluate the effect of airborne particulate matter 2.5 (PM2.5 in winter on airway inflammation, water-soluble supernatant (Sup and water-insoluble precipitate (Pre in PM2.5 were inoculated in NC/Nga mice with high sensitivity to mite allergens. Sup with aluminum oxide was injected intraperitoneally for sensitization. Five days later, Sup, Pre or both Sup and Pre were inoculated via the nasal route five times for more sensitization and a challenge inoculation on the 11th day in NC/Nga mice. On the 12th day, mice were examined for airway hyperresponsiveness (AHR, BALF cell count and IL-1β concentration, mRNA expression of Th1 and Th2 cytokines, chemokines such as eotaxin 1 and eotaxin 2, inflammasomal complex molecules such as IL-1β, caspase 1 and the nucleotide-binding domain and leucine-rich repeat protein 3 (NLRP3 in lung tissue as well as histopathology. The synergistic effect of Sup and Pre was observed in terms of increases in AHR, BALF cells, the mRNA expression of IL-13, eotaxin1 and IL-1β, and the IL-1β concentration in BALF. Intracellular deposits of insoluble particulates were observed in macrophages around inflammatory granulation of the mouse group treated with Sup and Pre. These results suggest that PM2.5 can induce airway hyperresponsiveness in mice with genetically high sensitivity to mite allergens by an inflammasome-associated mechanism and synergistic action of insoluble particulates and soluble components.

  3. Root inoculation with Pseudomonas putida KT2440 induces transcriptional and metabolic changes and systemic resistance in maize plants

    Directory of Open Access Journals (Sweden)

    Chantal ePlanchamp

    2015-01-01

    Full Text Available Pseudomonas putida KT2440 (KT2440 rhizobacteria colonize a wide range of plants. They have been extensively studied for their capacity to adhere to maize seeds, to tolerate toxic secondary metabolites produced by maize roots and to be attracted by maize roots. However, the response of maize plants to KT2440 colonization has not been investigated yet. Maize roots were inoculated with KT2440 and the local (roots and systemic (leaves early plant responses were investigated. The colonization behavior of KT2440 following application to maize seedlings was investigated and transcriptional analysis of stress- and defense-related genes as well as metabolite profiling of local and systemic maize tissues of KT2440-inoculated were performed. The local and systemic responses differed and more pronounced changes were observed in roots compared to leaves. Early in the interaction roots responded via jasmonic acid- and abscisic acid-dependent signaling. Interestingly, during later steps, the salicylic acid pathway was suppressed. Metabolite profiling revealed the importance of plant phospholipids in KT2440-maize interactions. An additional important maize secondary metabolite, a form of benzoxazinone, was also found to be differently abundant in roots three days after KT2440 inoculation. However, the transcriptional and metabolic changes observed in bacterized plants early during the interaction were minor and became even less pronounced with time, indicating an accommodation state of the plant to the presence of KT2440. Since the maize plants reacted to the presence of KT2440 in the rhizosphere, we also investigated the ability of these bacteria to trigger induced systemic resistance (ISR against the maize anthracnose fungus Colletotrichum graminicola. The observed resistance was expressed as strongly reduced leaf necrosis and fungal development in infected bacterized plants compared to non-bacterized controls, showing the potential of KT2440 to act as

  4. Seed quality, chlorophyll content index and leaf nitrogen levels in maize inoculated with Azospirillum brasilense

    Directory of Open Access Journals (Sweden)

    Lívia de Matos Pereira

    Full Text Available ABSTRACTThe aim of this study was to evaluate differences between maize genotypes in relation to the germination response of the seeds and the growth of seedlings inoculated with Azospirillum brasilense, as well as the effect of inoculation on nitrogen levels and the chlorophyll content index of the leaves. The physiological seeds quality from the single-cross hybrids AG7098 and 2B707, and from the experimental synthetic varieties V2 and V4, inoculated with A. brasilense, was tested for germination, percentage and rate of emergence, and dry matter weight of the shoots and roots. Nitrogen levels and chlorophyll content index were evaluated in leaves of the same four genotypes grown in a greenhouse under different nitrogen supply systems and methods of inoculation with A. brasilense. The genotypes differ with regard to inoculation with A. brasilense. The hybrids were responsive to inoculation with A. brasilense for root dry matter weight. The V2 variety had a lower performance as regards root dry matter weight, with the opposite being seen for the dry matter weight of the shoots. V4 displayed no significant differences when inoculated. The results of the chlorophyll content index were not significant. Each genotype under evaluation displayed a different response for leaf nitrogen levels. It is possible to infer that the hybrids responded better to inoculation with the bacteria, with the greater root development leading to a better utilisation of water and nutrients.

  5. Arbuscular mycorrhizal inoculation of peanut in low-fertile tropical soil : I. Host-fungus compatibility

    NARCIS (Netherlands)

    Quilambo, OA; Weissenhorn, I.; Kuiper, P.J C; Stulen, I.

    2005-01-01

    The effects of inoculation with an indigenous Mozambican and a commercial arbuscular mycorrhizal (AM) inoculant on two peanut (Arachis hypogaea L.) cultivars, a traditional, low-yielding Mozambican landrace (Local) and a modern, high-yielding cultivar (Falcon), were tested in a non-sterile and low-f

  6. Inoculation of Loblolly Pine Seedlings at Planting with Basidiospores of Ectomycorrhizal Fungi in Chip Form

    Science.gov (United States)

    Peter R. Beckjord; Marla S. McIntosh; Edward Hacskaylo; John H. Jr. Melhuish; John H. Jr. Melhuish

    1984-01-01

    Basidiospores of the ectomycorrhizae-forming fungi Pisolithus tinctorius and Scleroderma auranteum incorporated into an organic hydrocolloid can be used successfully in field inoculation. Containerized loblolly pine seedlings were inoculated during outplanting by this method. This study showed that basidiospore chips were effective inocula in this investigation.

  7. Silage extracts used to study the mode of action of silage inoculants in ruminants

    Science.gov (United States)

    Alfalfa and two corn crops were ensiled with and without Lactobacillus plantarum MTD/1 silage inoculant and fermented for 4 or 60 d to assess the effect of the inoculant on in vitro rumen fermentation of the resulting silages. Water and 80% ethanol extracts of the silages with added glucose were als...

  8. Silage inoculant effects on milk production and why that may be important to you

    Science.gov (United States)

    Silage inoculants are the most common additives used in making corn and hay crop silages. The main active ingredients in these products are lactic acid bacteria that help ensure the fermentation in the silo goes in a direction that helps preserve the crop. While inoculants have been available for ma...

  9. Milk production response to feeding alfalfa silage inoculated with Lactobacillus plantarum

    Science.gov (United States)

    In mini-silo trials, silages treated with a Lactobacillus plantarum silage inoculant (Ecosyl, Yorkshire, UK) had increased in vitro rumen microbial biomass production compared to untreated. Our objective was to determine if alfalfa silage treated with this inoculant could produce a milk production r...

  10. Response of Andean and Mesoamerican common bean genotypes to inoculation with rhizobium strains.

    Science.gov (United States)

    In most common bean (Phaseolus vulgaris L.) production regions of Latin America, inoculants are rarely used by farmers in spite of several studies that demonstrate the importance of Rhizobium inoculation on commercial production of legume crops. This study investigated specific bean host plant-Rhizo...

  11. Arbuscular mycorrhizal inoculation of peanut in low-fertile tropical soil. II. Alleviation of drought stress

    NARCIS (Netherlands)

    Quilambo, OA; Weissenhorn, I.; Doddema, H; Kuiper, PJC; Stulen, I.

    2005-01-01

    The effect of drought stress and inoculation with an indigenous Mozambican and a commercial arbuscular mycorrhizal (AM) inoculant on root colonization and plant growth and yield was studied in two peanut (Arachis hypogaea L.) cultivars-a traditional, low-yielding Mozambican landrace (Local) and a mo

  12. Hypothalamic-Pituitary-Adrenal Axis Physiology and Cognitive Control of Behavior in Stress Inoculated Monkeys

    Science.gov (United States)

    Parker, Karen J.; Buckmaster, Christine L.; Lindley, Steven E.; Schatzberg, Alan F.; Lyons, David M.

    2012-01-01

    Monkeys exposed to stress inoculation protocols early in life subsequently exhibit diminished neurobiological responses to moderate psychological stressors and enhanced cognitive control of behavior during juvenile development compared to non-inoculated monkeys. The present experiments extended these findings and revealed that stress inoculated…

  13. [The effect of soil inoculation with microbial pesticide destructors on plant growth and development].

    Science.gov (United States)

    Lisina, T O; Garan'kina, N G; Kruglov, Iu V

    2001-01-01

    Soil inoculation with liquid cultures of Bacillus megaterium 501 and Exophiala nigrum A-29 capable of degrading several organophosphorus pesticides accelerated growth and development of experimental plants, formation of their generative organs, and improved their productivity. This was particularly observed under stress plant growth conditions on phytotoxic peach substrates. The microorganisms inoculated can probably degrade phytotoxins present in soils, thereby favoring the plant development.

  14. Fermentative profile and bacterial diversity of corn silages inoculated with new tropical lactic acid bacteria.

    Science.gov (United States)

    Santos, A O; Ávila, C L S; Pinto, J C; Carvalho, B F; Dias, D R; Schwan, R F

    2016-02-01

    This study aimed to evaluate the effects of inoculation of strains of lactic acid bacteria (LAB) isolated from sugarcane grown in a Brazil on the quality of corn silage. Three strains of Lactobacillus buchneri (UFLA SLM11, UFLA SLM103 and UFLA SLM108), five strains of Lactobacillus plantarum (UFLA SLM08, UFLA SLM41, UFLA SLM45, UFLA SLM46 and UFLA SLM105), and one strain of Leuconostoc mesenteroides (UFLA SLM06) were evaluated at 0, 10, 30, 60 and 90 day after inoculating corn forage. The inoculation of the LAB strains did not influence the chemical composition of the silage, but pH, acetic acid and 1,2-propanediol were affected by treatment. The silages inoculated with UFLA SLM11 and SLM108 contained the lowest yeast and filamentous fungi counts during fermentation. Bacteria belonging to the Enterobacteriaceae family, Clostridium genus were detected in the silages inoculated with Lact. buchneri UFLA SLM 11, 103 and 108, as shown by DGGE analysis. Silages inoculated with Lact. buchneri UFLA SLM 11 showed higher aerobic stability. The Lact. buchneri UFLA SLM11 strain was considered promising as a starter culture or inoculant for corn silages. The selection of microbial inoculants for each crop promotes improvement of silage quality. Studies on the chemical and microbiological characteristics of silage provide useful information for improving ensiling techniques. © 2015 The Society for Applied Microbiology.

  15. Assessment of early blight (Alternaria solani) resistance in tomato using a droplet inoculation method

    NARCIS (Netherlands)

    Chaerani, R.; Groenwold, R.; Stam, P.; Voorrips, R.E.

    2006-01-01

    A droplet inoculation method was used for evaluation of tomato resistance to early blight, a destructive foliar disease of tomato caused by Alternaria solani (Ellis and Martin) Sorauer. In this test method, leaflets are inoculated with small droplets of a spore suspension in either water or a 0.1% a

  16. 9 CFR 113.37 - Detection of pathogens by the chicken embryo inoculation test.

    Science.gov (United States)

    2010-01-01

    ... embryo inoculation test. 113.37 Section 113.37 Animals and Animal Products ANIMAL AND PLANT HEALTH... VECTORS STANDARD REQUIREMENTS Standard Procedures § 113.37 Detection of pathogens by the chicken embryo...-serum mixture shall be inoculated into each of at least 20 fully susceptible chicken embryos. (1)...

  17. A Stress Inoculation Program for Parents Whose Children Are Undergoing Painful Medical Procedures.

    Science.gov (United States)

    Jay, Susan M.; Elliott, Charles H.

    1990-01-01

    Compared program efficacy in helping parents cope with children's painful medical procedures. Parents (n=72) of pediatric leukemia patients participated in either stress inoculation program or observed child participating in cognitive behavior therapy. Found parents in stress inoculation program reported lower anxiety scores and higher positive…

  18. Evaluation of spray and point inoculation methods for the phenotyping of Puccinia striiformis on wheat

    DEFF Research Database (Denmark)

    Sørensen, Chris Khadgi; Thach, Tine; Hovmøller, Mogens Støvring

    2016-01-01

    flexible application procedure for spray inoculation and it gave highly reproducible results for virulence phenotyping. Six point inoculation methods were compared to find the most suitable for assessment of pathogen aggressiveness. The use of Novec 7100 and dry dilution with Lycopodium spores gave...

  19. Gastric hyperplasia and parietal cell loss in Taenia taeniaeformis inoculated immunodeficient mice.

    Science.gov (United States)

    Lagapa, Jose Trinipil; Konno, Kenjiro; Oku, Yuzaburo; Nonaka, Nariaki; Ito, Mamoru; Kamiya, Masao

    2002-03-01

    Immunodeficient mice were studied to determine their suitability as models in investigating the role of Taenia taeniaeformis larval products in the development of gastric hyperplasia. Recombinant active gene 2 (RAG2)-deficient and severe combined immune-deficient (SCID) mice were studied as candidate animal models. RAG2-deficient mice inoculated orally with T. taeniaeformis eggs developed gastric hyperplasia with alcian blue-periodic acid-Schiff-positive cell proliferation similar to those of rats. SCID mice inoculated with different doses and routes of T. taeniaeformis in vitro-hatched oncospheres and those orally inoculated with eggs resulted also in different degrees of gastric hyperplasia. Influence of inoculation forms of parasite, doses and routes of inoculation on initiation of hyperplastic gastropathy was suggested to be dependent on number and size of developed larvae. Both RAG2-deficient and SCID mice with hyperplastic mucosa were observed with significant loss of parietal cells. Apparent decrease in parietal cell number was observed in SCID mice at 2 weeks after intraperitoneal inoculation with oncospheres before hyperplastic lesions developed. Earliest occurrence of gastric hyperplasia in SCID mice was observed at 3 weeks after oral inoculation of in vitro-hatched oncospheres, sooner than orally inoculated rats. The results suggested that these immunodeficient mice could be used as animal models to study factors involved in T. taeniaeformis-induced gastric mucous cell hyperplasia.

  20. Nitrogen translocation in wheat inoculated with Azospirillum and fertilized with nitrogen

    Directory of Open Access Journals (Sweden)

    RODRIGUES OSMAR

    2000-01-01

    Full Text Available The productivity and the translocation of assimilates and nitrogen (N were compared after inoculation of wheat (Triticum aestivum L., cv. BR-23 seeds with two strains of Azospirillum brasilense (strains 245 and JA 04 under field conditions. The inoculation of wheat seeds was done with a peat inoculant at sowing time. Plant material for evaluations were collected at anthesis and maturity. No differences in grain yield and in the translocation of assimilates resulting from inoculation were detected. Differences were observed in relation to N rates (0, 15, and 60 kg ha-1. N content in the grain increased significantly in the bacteria-inoculated treatments in which N was not added. This increase in N content in the grain with inoculation was probably due to higher N uptake after anthesis without any significant contribution on the grain yield. Such increment was of 8.4 kg ha-1 of N representing 66% more N than in no inoculated treatment. Regardless of the inoculation and the rate of N applied, it was observed that about 70% of the N accumulated at anthesis was translocated from vegetative parts to the grain.

  1. [Quantitative evaluation of printing accuracy and tissue surface adaptation of mandibular complete denture polylactic acid pattern fabricated by fused deposition modeling technology].

    Science.gov (United States)

    Deng, K H; Wang, Y; Chen, H; Zhao, Y J; Zhou, Y S; Sun, Y C

    2017-06-09

    Objective: To quantitatively evaluate the adaptation of polylactic acid (PLA) pattern of mandibular complete denture fabricated by fused deposition modeling (FDM) technology. Methods: A mandibular complete denture digital model was designed through a complete denture design software based on a pair of standard maxillomandibular edentulous plaster model and their occlusion bases. Ten PLA mandibular complete dentures were printed with a FDM machine. The dentures were scanned with and without the plaster model using a three-dimensional (3D) scanner. In Geomagic software, the scanning data of printed dentures were registered to its computer aided design (CAD) data, and the printing error was analyzed using the multipoint registration command. For quantitatively evaluating the adaptation of the denture, the data of plaster model and PLA denture were registered to the whole data of denture located in the plaster model using the best-fit alignment command, the 3D deviation of the plaster model and tissue surface of the denture represent the space between them. The overall area was separated into three parts: primary stress-bearing area, secondary stress-bearing area and border seal area, and the average deviations of these three parts were measured. The values were analyzed using analysis of variance. Results: Compared with the CAD data, the printing error was (0.013±0.004) mm. The overall 3D deviation between PLA denture and plaster model was (0.164±0.033) mm, in which the primary stress-bearing area was (0.165± 0.045) mm, the secondary stress-bearing area was (0.153 ± 0.027) mm, the border seal area was (0.186 ± 0.043) mm. These showed a good fit in the majority parts of the FDM denture to the plaster model. No statistically significant difference was observed between the three areas (F=1.857, P=0.175>0.05). Conclusions: Combined with the 3D scanning, CAD and FDM technology, a FDM 3D printing process of complete denture for injection moulding can be established. As

  2. In vivo adherence of Flavobacterium psychrophilum to mucosal external surfaces of rainbow trout (Oncorhynchus mykiss) fry

    DEFF Research Database (Denmark)

    Papadopoulou, A.; Dalsgaard, Inger; Lindén, A.

    2017-01-01

    and kidney were taken and inoculated onto TYES agar plates. Colony phenotypes of F. psychrophilum were identified and number of colonies counted. The results showed that cells of both phenotypes initially (0 h) adhered to all mucosal surfaces, but only the rough cells were still present on tissues 1 h post....... psychrophilum to mucosal tissues of rainbow trout fry and to test the skin mucus as a nutrient for the growth of F. psychrophilum. Fish were immersed in water containing 106  CFU  mL-1 F. psychrophilum for each colony phenotype. Mucosal tissue samples from fins, gills, skin and eyes, and swab samples from spleen...

  3. Ultrastructural observation of the airways of recovered and susceptible pigs after inoculation with Mycoplasma hyopneumoniae

    Directory of Open Access Journals (Sweden)

    Irigoyen Luiz Francisco

    1998-01-01

    Full Text Available To determine the morphological differences in the epithelium of the airways of recovered and susceptible pigs after Mycoplasma hyopneumoniae challenge, twenty-four 4-week-old M. hyopneumoniae-free pigs were intratracheally inoculated with 107ccu/ml of a pure low-passaged culture of the P5722-3 strain of M. hyopneumoniae challenge material. Eight pigs (group I were challenged at the beginning of the experiment and rechallenged 3 months later. Group II pigs were also challenged at the beginning of the experiment and necropsied 3 months later. Group III pigs were challenged at the same time as the rechallenge of group I pigs. Eight nonchallenged pigs served as controls (group IV. Three days after the second challenge of group I and the first challenge of group III, and every 3 and 4 days thereafter, two pigs from each group were euthanatized by electrocution and necropsied. Samples of bronchi and lung tissue were examined using light and electron microscopy (SEM and TEM. Macroscopic lesions were observed in the lungs of all group III pigs (average = 4.74% and were characterized by purple-red areas of discoloration and increased firmness affecting the cranioventral aspect of the lungs. Macroscopic lesions of pneumonia in groups I and II were minimal (less than 1%. There were no gross lesions of pneumonia in control (group IV pigs. Microscopic lesions were characterized by hyperplasia of the peribronchial lymphoid tissue and mild neutrophilic infiltrates in alveoli. Electron microscopy showed patchy areas with loss of cilia and presence of leukocytes and mycoplasmas in bronchi of susceptible pigs (group III. The bronchial epithelium of rechallenged (group I, recovered (group II, and control (group IV pigs was ultrastructurally similar indicating recovery of the former two groups. Although mycoplasmas were seen among cilia, a second challenge on pigs of group I did not produce another episode of the disease nor did it enhance morphological changes

  4. Assessment of Qualitative and Quantitative Characters of Two Persian Clover Ecotypes Inoculated by Rhizobium leguminosarum biovartrifoli and Pesudomonas putida Bacteria

    Directory of Open Access Journals (Sweden)

    R Azamei

    2016-02-01

    the powder used for inoculum was 5 gram from every bacterium seed (Containing 108 × 5 bacteria per gram for 100 kg seed. For better adhesion of the bacteria on the surface of seed, water with 20% sugar solution was used. (2 Sampling performed by harvesting 2 meter of every plot and 2 kg of fresh yield was transferred to oven (48 hours with (72 °C to determine dry matter. The crude protein content was determined by multiplying nitrogen content by the correction factor (6.25. At the end of the season a few plants from each plot were selected, and the number of nodules formed on the roots were counted and secondary roots with the highest penetration depth were measured by ruler. Analysis of variance and the means of treatments were performed using SAS software. Average interaction MSTAT- C treatments were performed using the software and the treatments were compared based on Duncan method. Results and Discussion The results showed that the qualitative factors have not been affected by Rhizobium and Pseudomonas bacteria so that the differences between treated plots with control were not significant. The quantitative factors except for depth of root penetration were affected by inoculation with Rhizobium and Pseudomonas bacteria and a significant difference between treated plots and control was obtained. Roberts et al. (3 reported that delay in planting caused plant growth exposure to high temperatures and long days, thus cell walls were stimulated to increase fiber and protein content and consequently digestibility was extremely reduced. The results indicated that inoculation by both bacteria (Rhizobium and Pseudomonas has affected quantitative indicators of growth significantly and increased fresh and dry yield, plant height, leaf/stem ratio and the number of nodes on roots. However the results showed that the qualitative characteristics and root infiltration did not change significantly by inoculation of both bacteria. Plant growth promoting rhizobacteria (PGPR leaded

  5. Atmospheric cold plasma inactivation of Escherichia coli, Salmonella enterica serovar Typhimurium and Listeria monocytogenes inoculated on fresh produce.

    Science.gov (United States)

    Ziuzina, D; Patil, S; Cullen, P J; Keener, K M; Bourke, P

    2014-09-01

    Atmospheric cold plasma (ACP) represents a potential alternative to traditional methods for non-thermal decontamination of foods. In this study, the antimicrobial efficacy of a novel dielectric barrier discharge ACP device against Escherichia coli, Salmonella enterica Typhimurium and Listeria monocytogenes inoculated on cherry tomatoes and strawberries, was examined. Bacteria were spot inoculated on the produce surface, air dried and sealed inside a rigid polypropylene container. Samples were indirectly exposed (i.e. placed outside plasma discharge) to a high voltage (70 kVRMS) air ACP and subsequently stored at room temperature for 24 h. ACP treatment for 10, 60 and 120 s resulted in reduction of Salmonella, E. coli and L. monocytogenes populations on tomato to undetectable levels from initial populations of 3.1, 6.3, and 6.7 log10 CFU/sample, respectively. However, an extended ACP treatment time was necessary to reduce bacterial populations attached on the more complex surface of strawberries. Treatment time for 300 s resulted in reduction of E. coli, Salmonella and L. monocytogenes populations by 3.5, 3.8 and 4.2 log10 CFU/sample, respectively, and also effectively reduced the background microflora of tomatoes.

  6. Measurement of binding of basic drugs to acidic phospholipids using surface plasmon resonance and incorporation of the data into mechanistic tissue composition equations to predict steady-state volume of distribution.

    Science.gov (United States)

    Small, Helen; Gardner, Iain; Jones, Hannah M; Davis, John; Rowland, Malcolm

    2011-10-01

    Acidic phospholipid binding plays an important role in determining the tissue distribution of basic drugs. This article describes the use of surface plasmon resonance to measure binding affinity (K(D)) of three basic drugs to phosphatidylserine, a major tissue acidic phospholipid. The data are incorporated into mechanistic tissue composition equations to allow prediction of the steady-state volume of distribution (V(ss)). The prediction accuracy of V(ss) using this approach is compared with the original methodology described by Rodgers et al. (J Pharm Sci 94:1259-1276), in which the binding to acidic phospholipids is calculated from the blood/plasma concentration ratio (BPR). The compounds used in this study [amlodipine, propranolol, and 3-dimethylaminomethyl-4-(4-methylsulfanyl-phenoxy)-benzenesulfonamide (UK-390957)] showed higher affinity binding to phosphatidylserine than to phosphatidylcholine. When the binding affinity to phosphatidylserine was incorporated into mechanistic tissue composition equations, the V(ss) was more accurately predicted for all three compounds by using the surface plasmon resonance measurement than by using the BPR to estimate acidic phospholipid binding affinity. The difference was particularly marked for UK-390957, a sulfonamide that has a high BPR due to binding to carbonic anhydrase. The novel approach described in this article allows the binding affinity of drugs to an acidic phospholipid (phosphatidylserine) to be measured directly and demonstrates the utility of the binding data in the prediction of V(ss).

  7. Selection of Infective Arbuscular Mycorrhizal Fungal Isolates for Field Inoculation

    Directory of Open Access Journals (Sweden)

    Elisa Pellegrino

    2010-09-01

    Full Text Available Arbuscular mycorrhizal (AM fungi play a key role in host plant growth and health, nutrient and water uptake, plant community diversity and dynamics. AM fungi differ in their symbiotic performance, which is the result of the interaction of two fungal characters, infectivity and efficiency. Infectivity is the ability of a fungal isolate to establish rapidly an extensive mycorrhizal symbiosis and is correlated with pre-symbiotic steps of fungal life cycle, such as spore germination and hyphal growth. Here, different AM fungal isolates were tested, with the aim of selecting infective endophytes for field inoculation. Greenhouse and microcosm experiments were performed in order to assess the ability of 12 AM fungal isolates to produce spores, colonize host roots and to perform initial steps of symbiosis establishment, such as spore germination and hyphal growth. AM fungal spore production and root colonization were significantly different among AM fungal isolates. Spore and sporocarp densities ranged from 0.8 to 7.4 and from 0.6 to 2.0 per gram of soil, respectively, whereas root colonization ranged from 2.9 to 72.2%. Percentage of spore or sporocarp germination ranged from 5.8 to 53.3% and hyphal length from 4.7 to 79.8 mm. The ordination analysis (Redundancy Analysis, RDA showed that environmental factors explained about 60% of the whole variance and their effect on fungal infectivity variables was significant (P = 0.002. The biplot clearly showed that variables which might be used to detect infective AM fungal isolates were hyphal length and root colonization. Such analysis may allow the detection of the best parameters to select efficient AM fungal isolates to be used in agriculture.

  8. Effect of the inoculation of a starter culture and vacuum packaging during the resting stage on sensory traits of dry-cured ham.

    Science.gov (United States)

    Sánchez-Molinero, F; Arnau, J

    2008-12-01

    The effects of the inoculation of a mixed starter culture and vacuum packaging (during resting stage) on odour, appearance, texture and flavour of dry-cured ham were studied. After salting, half of the 36 processed hams were inoculated with a commercial starter culture containing lactic-acid bacteria, Gram-positive catalase-positive cocci and yeasts. Nine hams per group (inoculated and non-inoculated) remained vacuum-packaged during resting. External odour during the process, as well as appearance of the cut surface, texture and flavour on semimembranosus and biceps femoris of the final product were assessed. Vacuum packaging during resting caused an increase in white film and feedstuff flavour, as well as a decrease in aged flavour, hardness, fibrousness and overall liking. The use of the starter culture brought about an increase in feedstuff flavour, a decrease in sweetness, aged flavour, nutty flavour and overall liking and, only in vacuum-packaged hams, the development of a floral flavour, but had no significant effect on texture descriptors. The starter culture studied is considered inappropriate for the production of traditional Spanish dry-cured ham regardless of the type of resting used.

  9. Feeding response of Ips paraconfusus to phloem and phloem metabolites of Heterobasidion annosum-inoculated ponderosa pine, Pinus ponderosa.

    Science.gov (United States)

    McNee, William R; Bonello, Pierluigi; Storer, Andrew J; Wood, David L; Gordon, Thomas R

    2003-05-01

    In studies of feeding by the bark beetle, Ips paraconfusus, two pine stilbenes (pinosylvin and pinosylvin methyl ether), ferulic acid glucoside, and enantiomers of the four most common sugars present in ponderosa pine phloem (sucrose, glucose, fructose, and raffinose) did not stimulate or reduce male feeding when assayed on wet alpha-cellulose with or without stimulatory phloem extractives present. When allowed to feed on wet alpha-cellulose containing sequential extracts (hexane, methanol, and water) of ponderosa pine phloem, methanol and water extractives stimulated feeding, but hexane extractives did not. Males confined in wet alpha-cellulose containing aqueous or organic extracts of culture broths derived from phloem tissue and containing the root pathogen. Heterobasidion annosum, ingested less substrate than beetles confined to control preparations. In an assay using logs from uninoculated ponderosa pines, the mean lengths of phloem in the digestive tracts increased as time spent feeding increased. Males confined to the phloem of basal logs cut from ponderosa pines artificially inoculated with H. annosum ingested significantly less phloem than beetles in logs cut from trees that were (combined) mock-inoculated or uninoculated and did not contain the pathogen. However, individual pathogen-containing treatments were not significantly different from uninoculated controls. It was concluded that altered feeding rates are not a major factor which may explain why diseased ponderosa pines are colonized by I. paraconfusus.

  10. Phytohormones and induction of plant-stress tolerance and defense genes by seed and foliar inoculation with Azospirillum brasilense cells and metabolites promote maize growth.

    Science.gov (United States)

    Fukami, Josiane; Ollero, Francisco Javier; Megías, Manuel; Hungria, Mariangela

    2017-12-01

    Azospirillum spp. are plant-growth-promoting bacteria used worldwide as inoculants for a variety of crops. Among the beneficial mechanisms associated with Azospirillum inoculation, emphasis has been given to the biological nitrogen fixation process and to the synthesis of phytohormones. In Brazil, the application of inoculants containing A. brasilense strains Ab-V5 and Ab-V6 to cereals is exponentially growing and in this study we investigated the effects of maize inoculation with these two strains applied on seeds or by leaf spray at the V2.5 stage growth-a strategy to relieve incompatibility with pesticides used for seed treatment. We also investigate the effects of spraying the metabolites of these two strains at V2.5. Maize growth was promoted by the inoculation of bacteria and their metabolites. When applied via foliar spray, although A. brasilense survival on leaves was confirmed by confocal microscopy and cell recovery, few cells were detected after 24 h, indicating that the effects of bacterial leaf spray might also be related to their metabolites. The major molecules detected in the supernatants of both strains were indole-3-acetic acid, indole-3-ethanol, indole-3-lactic acid and salicylic acid. RT-PCR of genes related to oxidative stress (APX1, APX2, CAT1, SOD2, SOD4) and plant defense (pathogenesis-related PR1, prp2 and prp4) was evaluated on maize leaves and roots. Differences were observed according to the gene, plant tissue, strain and method of application, but, in general, inoculation with Azospirillum resulted in up-regulation of oxidative stress genes in leaves and down-regulation in roots; contrarily, in general, PR genes were down-regulated in leaves and up-regulated in roots. Emphasis should be given to the application of metabolites, especially of Ab-V5 + Ab-V6 that in general resulted in the highest up-regulation of oxidative-stress and PR genes both in leaves and in roots. We hypothesize that the benefits of inoculation of Azospirillum on

  11. Population of Pratylenchus coffeae (Z. and growth of Arabica coffee seedling inoculated by Pseudomonas diminuta L. and Bacillus subtilis (C..

    Directory of Open Access Journals (Sweden)

    Irfan Fauzi

    2015-03-01

    Full Text Available AbstractPratylenchus coffeae is a parasitic nematoda that infected the roots of some plants, one of them is coffee. The Infection of Pratylenchus coffeae cause root tissue damage that led to root lession and make root become rotten, it will interfere the ability of roots to absorb water and nutrients in the soil which resulted in the growth of plants. At the moment, control of Pratylenchus coffeae are following integrated pests management (IPM program, which integrated between the use of coffee resistant clone and application of biological agents. Research on biological control was conducted more intensive, at the moment; due to it is friendlier save against environment and cheaper then using chemical nematicides. The research was conducted to know the effects of Micorrhiza Helper Bacteria (MHB,Pseudomonas diminuta and Bacillus subtilis in suppressing the population of P. coffeaeas well as their effect on growth of coffee seedling.  Coffee arabica (Coffea arabica L. seedling one moth old were used in the experiment. The experiment prepared with eight treatments and five  replications, as follows: A (Pseudomonas diminuta with density of 108 cfu / ml, B (Pseudomonas diminuta with density of 2x108 cfu / ml, C (Bacillus subtilis with density of 108 cfu / ml , D (Bacillus subtilis with density 2x108 cfu / ml, E (Carbofuran nematicide 5 g formulation / pot, F (Pseudomonas diminuta and Bacillus subtilis with each density of 108 cfu / ml, K- (Nematoda inoculation but without bacteria and nematicide, K+ (coffee seedling  without any additional treatment. The experiment was conducted for sixteen weeks or about four months. The results of the experiment showed that application of MHB could suppress population of P. coffeae and increase coffee seedling growth significantly. Inoculation of B. subtilis at 108 cfu per seedling suppressed significantly nematoda population of 71.3% compared with untreated seedling but inoculated with nematoda. It was not

  12. Kitchen Practices Used in Handling Broiler Chickens and Survival of Campylobacter spp. on Cutting Surfaces in Kampala, Uganda

    National Research Council Canada - National Science Library

    Irene Wanyenya; Charles Muyanja; George William Nasinyama

    2004-01-01

    .... The survival of Campylobacter spp. on kitchen cutting surfaces was determined by inoculating approximately 106 CFU of Campylobacter jejuni onto sterile plastic, wooden, and metal cutting boards...

  13. Tissue tests

    NARCIS (Netherlands)

    Sonneveld, C.; Voogt, W.

    2009-01-01

    Tissue tests are widely used in horticulture practice and have in comparison with soil or substrate testing advantages as well disadvantages in comparison with soil testing. One of the main advantages of tissue tests is the certainty that analysed nutrients in plant tissues are really present in the

  14. Tissue tests

    NARCIS (Netherlands)

    Sonneveld, C.; Voogt, W.

    2009-01-01

    Tissue tests are widely used in horticulture practice and have in comparison with soil or substrate testing advantages as well disadvantages in comparison with soil testing. One of the main advantages of tissue tests is the certainty that analysed nutrients in plant tissues are really present in the

  15. Chicken cecal microRNAs in the response to Campylobacter jejuni inoculation by Solexa sequencing.

    Science.gov (United States)

    Liu, Xiaoyi; Liu, Liying; Zhang, Maozhi; Wang, Huicui; Yang, Ning; Li, Xianyao

    2016-12-01

    Campylobacter jejuni (C. jejuni) is one of major foodborne pathogen that cause human diarrhea by consuming C. jejuni contaminated chicken products. MicroRNAs play an integral role in many different biological processes including bacteria and virus inoculation in chickens. In this study, we identified chicken miRNAs responding to C. jejuni inoculation through Solexa sequencing in the cecum. As a result, four miRNAs were significantly differentially expressed between inoculated and non-inoculated groups. There were 1,114 putative target genes regulated by those differentially expressed miRNAs predicted by miRanda, TargetScan, and miRTarget softwares. Functional analysis of those target genes showed that 113 gene ontology biological process terms and 14 pathways were significantly enriched. Hedgehog signaling pathway may contribute to chicken C. jejuni inoculation. MiR-155 played vital role in the C. jejuni inoculation. The result herein will lay the foundation for the further study of regulatory mechanism of chicken miRNAs in the response to C. jejuni inoculation. © 2016 Poultry Science Association Inc.

  16. Re-Thinking Anxiety: Using Inoculation Messages to Reduce and Reinterpret Public Speaking Fears

    Science.gov (United States)

    Jackson, Ben; Compton, Josh; Thornton, Ashleigh L.; Dimmock, James A.

    2017-01-01

    Inoculation theory offers a framework for protecting individuals against challenges to an existing attitude, belief, or state. Despite the prevalence and damaging effects of public speaking anxiety, inoculation strategies have yet to be used to help individuals remain calm before and during public speaking. We aimed to test the effectiveness of an inoculation message for reducing the onset of public speaking anxiety, and helping presenters interpret their speech-related anxiety more positively. Participants (Mage = 20.14, SD = 2.72) received either an inoculation (n = 102) or control (n = 128) message prior to engaging a public speaking task and reported a range of anxiety-related perceptions. Accounting for personality characteristics and perceptions of task importance, and relative to control participants, those who received the inoculation message reported significantly lower pre-task anxiety, and following the task, reported that they had experienced lower somatic anxiety, and that the inoculation message had caused them to view their nerves in a less debilitating light. Inoculation messages may be an effective strategy for helping participants reframe and reduce their apprehension about public speaking, and investigating their efficacy in other stress-inducing contexts may be worthwhile. PMID:28125618

  17. Striatal dopamine D2/3 receptor regulation by stress inoculation in squirrel monkeys

    Directory of Open Access Journals (Sweden)

    Alex G. Lee

    2016-06-01

    Full Text Available Intermittent mildly stressful situations provide opportunities to learn, practice, and improve coping in a process called stress inoculation. Stress inoculation also enhances cognitive control and response inhibition of impulsive motivated behavior. Cognitive control and motivation have been linked to striatal dopamine D2 and/or D3 receptors (DRD2/3 in rodents, monkeys, and humans. Here, we study squirrel monkeys randomized early in life to stress inoculation with or without maternal companionship and a no-stress control treatment condition. Striatal DRD2/3 availability in adulthood was measured in vivo by [11C]raclopride binding using positron emission tomography (PET. DRD2/3 availability was greater in caudate and putamen compared to ventral striatum as reported in PET studies of humans and other non-human primates. DRD2/3 availability in ventral striatum was also consistently greater in stress inoculated squirrel monkeys compared to no-stress controls. Squirrel monkeys exposed to stress inoculation in the presence of their mother did not differ from squirrel monkeys exposed to stress inoculation without maternal companionship. Similar effects in different social contexts extend the generality of our findings and together suggest that stress inoculation increases striatal DRD2/3 availability as a correlate of cognitive control in squirrel monkeys.

  18. Improvement of Rhizobial Inoculants: A Key Process in Sustainable Soybean Production

    Directory of Open Access Journals (Sweden)

    Mihaela Blažinkov

    2016-01-01

    Full Text Available Symbiotic nitrogen fixation has important role in sustainable soybean production because of utilization of atmospheric nitrogen for soybean nutrition. Pre-sowing soybean seed inoculation with selected rhizobial strains is used to improve the amount of symbiotically fixed nitrogen. Besides strain selection, suitable inoculant formulation is important for the success of inoculant application. The aim of this research is the evaluation of symbiotic efficiency and compatibility of Bradyrhizobium japonicum strains with soybean cultivar as well as possibility of using different inoculant formulation in soybean production. During two years of field trials in eastern Slavonia, nodule dry weight, nitrogen content in plant, seed yield, 1000 seeds weight, protein and oil content in seed were determined. Results of this study indicate that inoculant formulation as well as the use of selected strains affects nodulation, symbiotic and agronomic properties of soybean. Despite the differences in results in both experimental years, it can be concluded that the strains used as well as inoculant formulations are suitable for soybean inoculation in agroecological conditions of eastern Slavonia.

  19. Combination of inoculation methods of Azospirilum brasilense with broadcasting of nitrogen fertilizer increases corn yield

    Directory of Open Access Journals (Sweden)

    Tânia Maria Müller

    2015-01-01

    Full Text Available Nitrogen (N is the most limiting nutrient for corn production. Thereby, the goal of the paper was to evaluate inoculation methods of Azospirillum brasilense in order to partially supply N required by the crop. The experiment was carried out in Guarapuava, PR, Brasil, in 2011/2012 growing season. Randomized blocks with factorial 3 inoculation methods (seed treatment, planting furrow and non-inoculated control x 5 doses of nitrogen (0, 75, 150, 225 and 300kg ha-1 x 8 replications was used as the experimental design. Leaf are index, foliar nitrogen content, total chlorophyll, grains per ear and yield were evaluated. There was significant interaction between inoculation methods and nitrogen fertilization to leaf area index, but not for yield. Inoculation with the diazotrophic bacteria provided yield increase of 702kg ha-1 for inoculation in seeding furrow and 432kg ha-1 for inoculation in seed treatment compared to the control, but both treatments did not differ between each other. Furthermore, total chlorophyll, grains per ear and yield were positively affected, with quadratic response, by the nitrogen fertilization in broadcasting

  20. [Queries related to the technology of soybean seed inoculation with Bradyrhizobium spp].

    Science.gov (United States)

    Lodeiro, Aníbal R

    2015-01-01

    With the aim of exploiting symbiotic nitrogen fixation, soybean crops are inoculated with selected strains of Bradyrhizobium japonicum, Bradyrhizobium diazoefficiens or Bradyrhizobium elkanii (collectively referred to as Bradyrhizobium spp.). The most common method of inoculation used is seed inoculation, whether performed immediately before sowing or using preinoculated seeds or pretreated seeds by the professional seed treatment. The methodology of inoculation should not only cover the seeds with living rhizobia, but must also optimize the chances of these rhizobia to infect the roots and nodulate. To this end, inoculated rhizobia must be in such an amount and condition that would allow them to overcome the competition exerted by the rhizobia of the allochthonous population of the soil, which are usually less effective for nitrogen fixation and thus dilute the effect of inoculation on yield. This optimization requires solving some queries related to the current knowledge of seed inoculation, which are addressed in this article. I conclude that the aspects that require further research are the adhesion and survival of rhizobia on seeds, the release of rhizobia once the seeds are deposited in the soil, and the movement of rhizobia from the vicinity of the seeds to the infection sites in the roots. Copyright © 2015 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  1. Co-inoculation with diazotrophic bacteria in soybeans associated to urea topdressing

    Directory of Open Access Journals (Sweden)

    Glauber Monçon Fipke

    Full Text Available ABSTRACT Increased grain yield can be obtained via an interaction between plants and growth-promoting microorganisms. The Bradyrhizobium spp. are capable of fixing atmospheric nitrogen in soybeans [Glycine max (L. Merril], and Azospirillum spp. induce the synthesis of phytohormones. The aim of this study was to evaluate inoculation with Bradyrhizobium and co-inoculation with Bradyrhizobium + Azospirillum brasilense in soybeans in combination with the application a topdressing of 0, 75 or 150 kg of N ha-1 of urea during the reproductive stage. Three soybean cultivars (BMX Ativa, TEC 6029 and BMX Potência, were tested in field experiments in Santa Maria, RS, Brazil, during two agricultural years (2013/2014 and 2014/2015 and two sowing times. Morphological, nodulation and yield components were evaluated. Co-inoculation increased the grain yield by 240 kg ha-1 compared with conventional inoculation. When co-inoculated, cultivars BMX Ativa, TEC 6029 and BMX Potência showed increased grain yields of 6, 4 and 12%, respectively. The application of 150 kg ha-1 of N as a topdressing increased the grain yield by 300 kg ha-1 in the co-inoculated cultivars TEC 6029 and BMX Potência, but without a financial return. When inoculated only with Bradyrhizobium, the cultivars did not respond positively to the application of urea.

  2. Enhanced phytoremediation of toxic metals by inoculating endophytic Enterobacter sp. CBSB1 expressing bifunctional glutathione synthase.

    Science.gov (United States)

    Qiu, Zhiqi; Tan, Hongming; Zhou, Shining; Cao, Lixiang

    2014-02-28

    To engineer plant-bacteria symbionts for remediating complex sites contaminated with multiple metals, the bifunctional glutathione (GSH) synthase gene gcsgs was introduced into endophytic Enterobacter sp. CBSB1 to improve phytoremediation efficiency of host plant Brassica juncea. The GSH contents of shoots inoculated with CBSB1 is 0.4μMg(-1) fresh weight. However, the GSH concentration of shoots with engineered CBSB1-GCSGS increased to 0.7μMg(-1) fresh weight. The shoot length, fresh weight and dry weight of seedlings inoculated with CBSB1-GCSGS increased 67%, 123%, and 160%, compared with seedlings without inoculation, respectively. The Cd and Pb concentration in shoots with CBSB1-GCSGS increased 48% and 59% compared with seedlings without inoculation, respectively. The inoculation of CBSB1 and CBSB1-GCSGS could increase the Cd and Pb extraction amounts of seedlings significantly compared with those without inoculation (PEnterobacter sp. CBSB1 upgraded the phytoremediation efficacy of B. juncea. So the engineered Enterobacter sp. CBSB1-GCSGS showed potentials in remediation sites contaminated with complex contaminants by inoculating into remediating plants.

  3. Effect of Inoculating Lactic Acid Bacteria Starter in Low-Salt Pickle Process of Zhacai

    Directory of Open Access Journals (Sweden)

    Shiyang Gao

    2012-10-01

    Full Text Available The traditional pickle process of Zhacai was thought unsafely and unstable, because of the high salt and nitrite contents. The study has shown a method to solve these problems. One Lactic Acid Bacteria (LAB strain, Lactobacillus plantarum was used as starter in the low salt pickle process of Chinese Zhacai. The LAB inoculation amounts were 2, 5, 8%, respectively and the group without LAB inoculation was set as the control. The inoculation groups were added with 8% salt, while the control group 10%. The dynamic changes of physical and chemical parameters of two groups were detected during the pickle process. The results showed that: pH value of the inoculated groups declined rapidly, the lowest pH value was 3.61; the LAB became the predominant bacteria strains during the pickle and the colony amounts of the inoculated groups could be up to 6.32 log10 cfu/mL; the nitrite content of each group kept rising and the nitrite peaks of the inoculated groups appeared 3 days in advance, but the peak values were nearly half of the control group; amino nitrogen contents were lower but finally higher than control group and the highest value could reach 0.46 g/100 mL finally. Therefore, the inoculated groups with the lower salt could be more safety and better flavor. This study provided important reference to optimize the pickle fermentation process of Chinese Zhacai.

  4. Re-Thinking Anxiety: Using Inoculation Messages to Reduce and Reinterpret Public Speaking Fears.

    Science.gov (United States)

    Jackson, Ben; Compton, Josh; Thornton, Ashleigh L; Dimmock, James A

    2017-01-01

    Inoculation theory offers a framework for protecting individuals against challenges to an existing attitude, belief, or state. Despite the prevalence and damaging effects of public speaking anxiety, inoculation strategies have yet to be used to help individuals remain calm before and during public speaking. We aimed to test the effectiveness of an inoculation message for reducing the onset of public speaking anxiety, and helping presenters interpret their speech-related anxiety more positively. Participants (Mage = 20.14, SD = 2.72) received either an inoculation (n = 102) or control (n = 128) message prior to engaging a public speaking task and reported a range of anxiety-related perceptions. Accounting for personality characteristics and perceptions of task importance, and relative to control participants, those who received the inoculation message reported significantly lower pre-task anxiety, and following the task, reported that they had experienced lower somatic anxiety, and that the inoculation message had caused them to view their nerves in a less debilitating light. Inoculation messages may be an effective strategy for helping participants reframe and reduce their apprehension about public speaking, and investigating their efficacy in other stress-inducing contexts may be worthwhile.

  5. Effects of Funnelliformis mosseae inoculation on alleviating atrazine damage in Canna indica L. var. flava Roxb.

    Science.gov (United States)

    Dong, Jing; Wang, Li; Ma, Fang; Yang, Jixian; Zhang, Xue; Zhao, Ting; Qi, Shanshan

    2017-01-02

    Atrazine residue in the environment continually damages plants and therefore requires immediate attention and effective development of methods for its decontamination. The effects of Funnelliformis mosseae inoculation on growth and physiology in atrazine-treated Canna indica L. var. flava Roxb. were investigated. At atrazine concentrations up to 15 mg L(-1), the growth of C. indica plants were negatively affected. Inoculation with F. mosseae alleviated the atrazine inhibition of plant growth and biomass. Furthermore, the chlorophyll content and root function increased under F. mosseae inoculation, and the oxidative stress of malondialdehyde, peroxidase, and superoxide dismutase activities induced by atrazine were also alleviated by F. mosseae inoculation. The removal rate of atrazine by untreated C. indica was significant, with removal rates of 20.5-55.3% by the end of a 14-day experiment; however, F. mosseae inoculation increased the removal rate to 35.6-75.1%. In conclusion, F. mosseae inoculation can alleviate the damage induced by atrazine in C. indica. Accordingly, C. indica inoculated with F. mosseae has excellent potential to be used in phytoremediation in habitats polluted by high atrazine concentrations.

  6. Alterations in plant growth and in root hormone levels of lodgepole pines inoculated with rhizobacteria.

    Science.gov (United States)

    Bent, E; Tuzun, S; Chanway, C P; Enebak, S

    2001-09-01

    The presence of other soil microorganisms might influence the ability of rhizobacterial inoculants to promote plant growth either by reducing contact between the inoculant and the plant root or by interfering with the mechanism(s) involved in rhizobacterially mediated growth promotion. We conducted the following experiments to determine whether reductions in the extent of growth promotion of lodgepole pine mediated by Paenibacillus polymyxa occur in the presence of a forest soil isolate (Pseudomonas fluorescens M20) and whether changes in plant growth promotion mediated by P. polymyxa (i) are related to changes in P. polymyxa density in the rhizosphere or (ii) result from alterations in root hormone levels. The extent of plant growth, P. polymyxa rhizosphere density, and root hormone concentrations were determined for lodgepole pine treated with (i) a single growth-promoting rhizobacterial strain (P. polymyxa L6 or Pw-2) or (ii) a combination of bacteria: strain L6 + strain M20 or strain Pw-2 + strain M20. There was no difference in the growth of pines inoculated with strain L6 and those inoculated with strain L6 + strain M20. However, seedlings inoculated with strain Pw-2 had more lateral roots and greater root mass at 12 weeks after inoculation than plants inoculated with strain Pw-2 + strain M20. The extent of growth promotion mediated by P. polymyxa L6 and Pw-2 in each treatment was not correlated to the average population density of each strain in the rhizosphere. Bacterial species-specific effects were observed in root hormone levels: indole-3-acetic acid concentration was elevated in roots inoculated with P. polymyxa L6 or Pw-2, while dihydrozeatin riboside concentration was elevated in roots inoculated with P. fluorescens M20.

  7. Effect of leachate recycling and inoculation on the biochemical characteristics of municipal refuse in landfill bioreactors

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Activity development of key groups of enzymes involved in municipal refuse decomposition was measured in laboratory landfill hioreactors with and without leachate recycling and inoculation for about 210 days.The results showed that the enzymes (amylase,protease,cellulase,lipase and pectinase ) were present in fresh refuse but at low values and positively affected hy leachate recycling and refuse inoculation.The total average of cellulase activity in digesters D3 operated with leachate recycling but no inoculation,D4 and D5 operated with leachate recycling and inoculation was much higher than that in digesters D1 and D2 without leachate recycling and inoculation by 88%-127%,117%-162% and 64%-98%.The total average of protease activity was higher in digester D4 than that in digesters D1,D2,D3and D5 by 63%,39%,24% and 24%,respectively,and the positive effect of leachate recycli.ng and inoculation on protease activity of landfilled refuse mainly was at the first two months.The total average of amylase activity was higher in digesters D3,D4 and D5 than that in digesters D1 and D2 by 83%-132%,96%-148% and 81%-129%.During the early phase of incubation,the stimulatory effect of inoculation on lipase activity was measured,but refuse moisture was the main factor affecting lipase activity of landfilled refuse.The inoculation,initial and continuous inoculation of microorganisms existing in leachate,was the mainly stimulatory factor affecting pectinase activity of landfilled refuse.

  8. Effect of Bacterial-Feeding Nematode Inoculation on{1mmWheatGrowth and N and P Uptake

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    A 40-day gnotobiotic microcosm experiment was carried out to quantifythe effect of bacterial-feeding nematode on plant growth and nutrientabsorption. The results showed that inoculation of bacterial-feedingnematode { Protorhabditis sp. stimulated the growth of wheat(Triticum aestivum) and the uptake of N. By the end of the40-day incubation wheat biomass and N uptake in the treatment withnematode and bacteria (Pseudomonas sp.) increased by 6.5% and5.9%, respectively, compared with bacteria alone treatment. Thepresence of nematode mainly accelerated the growth of aboveground ofwheat, while it slightly inhibited the root development. There waslittle difference in plant tissue N concentration between treatments. Pconcentration and uptake of wheat, however, were generally reduced bynematode. It appears that the enhancement of plant growth and nitrogenuptake is attributed to the enhancement of nitrogen mineralizationinduced by nematode feeding on bacteria, and the reduction ofphosphorous uptake is the result of weak root status and competition bybacteria immobilization.

  9. Improving Acacia auriculiformis seedlings using microbial inoculant (Beneficial Microorganisms)

    Institute of Scientific and Technical Information of China (English)

    Bayezid M. Khan; M.K. Hossain; M.A.U. Mridha

    2014-01-01

    A microbial inoculant, known as effective microorganisms (EM), was applied to determine its efficacy on seed germination and seedling growth in the nursery of Acacia auriculiformis A Cunn. ex Benth. The seedlings were grown in a mixture of sandy soil and cow dung (3:1) and kept in polybags;EM was poured at different concentra-tions (0.1%, 0.5%, 1%, 2%, 5% and 10%). Seed germination rate and growth parameters of seedlings - shoot and root lengths, fresh and dry weights of shoots and roots, vigor, volume, and quality indices and stur-diness - were measured. The nodulation status influenced by EM was also observed, along with the measurement of pigment contents in leaves. The highest germination rate (72%) was observed in 2% EM solution while the lowest (55%) was found in control treatment. The highest shoot and root lengths (30.6 cm and 31.2 cm respectively) were recorded in 2%EM and were significantly (p <0.05) different from control. Both fresh and dry weights of shoots were maximum (8.66 g and 2.99 g respectively) in 2% EM, whereas both fresh and dry weights of root were maximum (2.56 g and 1.23 g respectively) in 5%EM solution. Although the highest vigor index, volume index, and sturdiness (4450, 628 and 67.5 respec-tively) were found in 2% EM, the highest quality index (0.455) was found in 5%EM solution. The nodule number was higher at a very low (0.5%) concentration of EM but it normally decreased with the increase of concentration. The contents of chlorophyll a, chlorophyll b, and caro-tenoid were maximum (43.26 mg⋅L-1, 13.56 mg⋅L-1and 17.99 mg⋅L-1 respectively) in 2%EM. Therefore, low concentration of EM (up to 2%)can be recommended for getting maximum seed germination and seed-ling development of A. auriculiformis in the nursery.

  10. Liquid Microjunction Surface Sampling Coupled with High-Pressure Liquid Chromatography-Electrospray Ionization-Mass Spectrometry for Analysis of Drugs and Metabolites in Whole-Body Thin Tissue Sections

    Energy Technology Data Exchange (ETDEWEB)

    Kertesz, Vilmos [ORNL; Van Berkel, Gary J [ORNL

    2010-01-01

    In this work, a commercially available autosampler was adapted to perform direct liquid microjunction (LMJ) surface sampling followed by a high-pressure liquid chromatography (HPLC) separation of the extract components and detection with electrospray ionization mass spectrometry (ESI-MS). To illustrate the utility of coupling a separation with this direct liquid extraction based surface sampling approach, four different organs (brain, lung, kidney, and liver) from whole-body thin tissue sections of propranolol dosed and control mice were examined. The parent drug was observed in the chromatograms of the surface sampling extracts from all the organs of the dosed mouse examined. In addition, two isomeric phase II metabolites of propranolol (an aliphatic and an aromatic hydroxypropranolol glucuronide) were observed in the chromatograms of the extracts from lung, kidney, and liver. Confirming the presence of one or the other or both of these glucuronides in the extract from the various organs was not possible without the separation. These drug and metabolite data obtained using the LMJ surface sampling/HPLC-MS method and the results achieved by analyzing similar samples by conventional extraction of the tissues and subsequent HPLC-MS analysis were consistent.

  11. Canthaxanthin biosynthesis by Dietzia natronolimnaea HS-1: effects of inoculation and aeration rate

    Directory of Open Access Journals (Sweden)

    Forouzan Rostami

    2014-06-01

    Full Text Available The interest in production of natural colorants by microbial fermentation has been currently increased. The effects of D-glucose concentration (3.18-36.82 g/L, inoculum size (12.5 x 10(9-49.5 x 10(9 cfu cells/mL and air-flow rate (1.95-12.05 L/L min on the biomass, total carotenoid and canthaxanthin (CTX accumulation of Dietzia natronolimnaea HS-1 in a batch bioreactor was scrutinized using a response surface methodology-central composite rotatable design (RSM-CCRD. Second-order polynomial models with high R² values ranging from 0.978 to 0.990 were developed for the studied responses using multiple linear regression analysis. The models showed the maximum cumulative amounts of biomass (7.85 g/L, total carotenoid (5.48 mg/L and CTX (4.99 mg/L could be achieved at 23.38 g/L of D-glucose, 31.2 x 10(9 cfu cells/mL of inoculation intensity and air-flow rate of 7.85 L/L min. The predicted values for optimum conditions were in good agreement with experimental data.

  12. Effect of chlorine, blanching, freezing, and microwave heating on Cryptosporidium parvum viability inoculated on green peppers.

    Science.gov (United States)

    Duhain, G L M C; Minnaar, A; Buys, E M

    2012-05-01

    Cryptosporidium parvum oocysts have been found on the surface of vegetables in both developed and developing countries. C. parvum can contaminate vegetables via various routes, including irrigation water. This study investigated the effect of individual treatments of chlorine, blanching, blast freezing, and microwave heating, as well as combined treatments of chlorine and freezing, and chlorine and microwave heating on the viability of C. parvum oocysts inoculated on green peppers. The viability of the oocysts after the treatments was assessed using propidium iodide and a flow cytometer. Based on the propidium iodide staining, the chlorine treatments did not affect the viability of the oocysts. Blast freezing significantly inactivated 20% of the oocysts. Microwave heating and blanching significantly inactivated 93% of oocysts. Treatment with chlorine followed by blast freezing did not affect the viability of the oocysts significantly. Treatment with chlorine and microwave heating was significantly more effective than microwave heating alone and inactivated 98% of the oocysts. The study indicates that C. parvum oocysts are sensitive to heat and, to some extent, to blast freezing, but are resistant to chlorine. Therefore, the use of chlorine during vegetable processing is not a critical control point for C. parvum oocysts, and the consumption of raw or minimally processed vegetables may constitute a health risk as C. parvum oocysts can still be found viable on ready-to-eat, minimally processed vegetables.

  13. Growth response and nutrient uptake of blue pine (Pinus wallichiana seedlings inoculated with rhizosphere microorganisms under temperate nursery conditions

    Directory of Open Access Journals (Sweden)

    M.A. Ahangar

    2012-11-01

    Full Text Available Microbial inoculants (Trichoderma harzianum, Pseudomonas fluorescens,Laccaria laccata inoculated either individually or in combinationsignificantly improved the growth and biomass of blue pine seedlings. The ECM fungus Laccaria laccata, when inoculated individually, showed significantly higher plant growth, followed by Pseudomonas fluorescens and Trichoderma harzianum. The combined inoculation of rhizosphere microorganisms showed synergistic growth promoting action and proved superior in enhancing the growth of blue pine than individual inoculation. Co-inoculation of L. laccata with P. fluorescens resulted in higher ectomycorrhizal root colonization. Uptake of nutrients (N, P, K was significantly improved by microbial inoculants, tested individually or in combination. Combined inoculation of L. laccata with T. harzianum and P. fluorescens significantly increased in N, P and K contents in blue pine seedlings as compared to control. Acid phosphatase activity in the rhizosphere of blue pine seedlings was also enhanced by these microorganisms. L. laccata exhibited higher acid phosphatase activity followed by P. fluorescens.

  14. Baby bottle steam sterilizers for disinfecting home nebulizers inoculated with non-tuberculous mycobacteria.

    Science.gov (United States)

    Towle, D; Callan, D A; Lamprea, C; Murray, T S

    2016-03-01

    Non-tuberculous mycobacteria (NTMb), present in environmental water sources, can contribute to respiratory infection in patients with chronic pulmonary disease. Contaminated nebulizers are a potential source of respiratory infection. Treatment with baby bottle steam sterilizers disinfects home nebulizers inoculated with bacterial pathogens but whether this method works for disinfection of NTMb is unclear. Baby bottle steam sterilization was compared with vigorous water washing for disinfecting home nebulizers inoculated with NTMb mixed with cystic fibrosis sputum. No NTMb was recovered from any nebulizers after steam treatment whereas viable NTMb grew after water washing, demonstrating that steam sterilization effectively disinfects NTMb-inoculated nebulizers.

  15. Effect of bacterial inoculation in some mixtures of grassland legumes and gramineae

    Directory of Open Access Journals (Sweden)

    Claudiu Ghiocel

    2014-05-01

    Full Text Available Normal 0 false false false EN-US X-NONE X-NONE MicrosoftInternetExplorer4 The paper presents the influence of bacterial inoculation of perennial legumes (alfalfa, bird’s-foot trefoil in pure culture and cultivated in association with perennial gramineae on fodder yield, fodder quality, and atmospheric nitrogen-fixing ability. Results point out an increase of 10-12% of the yield of dry matter in the variants inoculated. Bacterial inoculation with specific bacterial stems (Sinorhizobium meliloti and Mezorhizobium loti has a positive influence on fodder quality too, materialised in the raw protein content and the amount of nitrogen fixed biologically.

  16. Differential response of kabuli and desi chickpea genotypes toward inoculation with PGPR in different soils

    Science.gov (United States)

    Imran, Asma; Mirza, Muhammad S.; Shah, Tariq M.; Malik, Kauser A.; Hafeez, Fauzia Y.

    2015-01-01

    Pakistan is among top three chickpea producing countries but the crop is usually grown on marginal lands without irrigation and fertilizer application which significantly hampers its yield. Soil fertility and inoculation with beneficial rhizobacteria play a key role in nodulation and yield of legumes. Four kabuli and six desi chickpea genotypes were, therefore, evaluated for inoculation response with IAA-producing Ochrobactrum ciceri Ca-34T and nitrogen fixing Mesorhizobium ciceri TAL-1148 in single and co-inoculation in two soils. The soil type 1 was previously unplanted marginal soil having low organic matter, P and N contents compared to soil type 2 which was a fertile routinely legume-cultivated soil. The effect of soil fertility status was pronounced and fertile soil on average, produced 31% more nodules, 62% more biomass and 111% grain yield than marginal soil. Inoculation either with O. ciceri alone or its co-inoculation with M. ciceri produced on average higher nodules (42%), biomass (31%), grains yield (64%) and harvest index (72%) in both chickpea genotypes over non-inoculated controls in both soils. Soil 1 showed maximum relative effectiveness of Ca-34T inoculation for kabuli genotypes while soil 2 showed for desi genotypes except B8/02. Desi genotype B8/02 in soil type 1 and Pb-2008 in soil type 2 showed significant yield increase as compared to respective un-inoculated controls. Across bacterial inoculation treatments, grain yield was positively correlated to growth and yield contributing parameters (r = 0.294* to 0.838*** for desi and r = 0.388* to 0.857** for kabuli). PCA and CAT-PCA analyses clearly showed a site-specific response of genotype x bacterial inoculation. Furthermore, the inoculated bacterial strains were able to persist in the rhizosphere showing colonization on root and within nodules. Present study shows that plant growth promoting rhizobacteria (PGPR) inoculation should be integrated with national chickpea breading program in

  17. Micro pore arrays in free standing cyclic olefin copolymer membranes: fabrication and surface functionalization strategies for in-vitro barrier tissue models

    Science.gov (United States)

    Gel, M.; Kandasamy, S.; Cartledge, K.; Be, C. L.; Haylock, D.

    2013-12-01

    In recent years there has been growing interest in micro engineered in-vitro models of tissues and organs. These models are designed to mimic the in-vivo like physiological conditions with a goal to study human physiology in an organ-specific context or to develop in-vitro disease models. One of the challenges in the development of these models is the formation of barrier tissues in which the permeability is controlled locally by the tissues cultured at the interface. In-vitro models of barrier tissues are typically created by generating a monolayer of cells grown on thin porous membranes. This paper reports a robust preparation method for free standing porous cyclic olefin copolymer (COC) membranes. We also demonstrate that gelatin coated membranes facilitate formation of highly confluent monolayer of HUVECs. Membranes with thickness in the range of 2-3 um incorporating micro pores with diameter approximately 20 um were fabricated and integrated with microfluidic channels. The performance of the device was demonstrated with a model system mimicking the endothelial barrier in bone marrow sinusoids.

  18. Behaviour of co-inoculated pathogenic and spoilage bacteria on poultry following several decontamination treatments.

    Science.gov (United States)

    Alonso-Hernando, Alicia; Capita, Rosa; Alonso-Calleja, Carlos

    2012-10-01

    The potential of chemical decontaminants to cause harmful effects on human health is among the causes of the rejection of antimicrobial treatments for removing surface contamination from poultry carcasses in the European Union. This study was undertaken to determine whether decontaminants might give a competitive advantage to pathogenic bacteria on poultry and involve a potential risk to consumer. A total of 144 chicken legs were co-inoculated with similar concentrations of pathogenic bacteria (Listeria monocytogenes, Staphylococcus aureus, Salmonella enterica serotype Enteritidis or Escherichia coli) and spoilage bacteria (Brochothrix thermosphacta or Pseudomonas fluorescens). Samples were dipped for 15min in solutions (w/v) of trisodium phosphate (12%; TSP), acidified sodium chlorite (1200ppm; ASC), citric acid (2%; CA), peroxyacids (220ppm; PA) or chlorine dioxide (50ppm; CD), or were left untreated (control). Microbiological analyses were carried out on day 0 and every 24h until day 7 of storage (at 10±1°C). The modified Gompertz equation was used as the primary model to fit observed data. TSP, ASC and CA were effective in extending the lag phase (L, ranging from 1.47±1.34days to 4.06±1.16days) and in decreasing the concentration of bacteria during the stationary phase (D, ranging from 2.46±0.51 log(10) cfu/cm(2) to 8.64±0.53 log(10) cfu/cm(2)), relative to the control samples (L values ranging from 0.59±0.38days and 2.52±2.28days, and D values ranging from 6.32±0.89 log(10) cfu/cm(2) to 9.39±0.39 log(10) cfu/cm(2), respectively). Both on untreated and on most decontaminated samples the overgrowth of spoilage bacteria among the species tested was observed throughout storage, suggesting that spoilage would occur prior to any noteworthy increase in the levels of pathogenic microorganisms. However, L. monocytogenes counts similar to, or higher than, those for spoilage bacteria were observed on samples treated with TSP, ASC or CA, suggesting that these

  19. Intracerebral inoculation of mouse-passaged Saffold virus type 3 affects cerebellar development in neonatal mice.

    Science.gov (United States)

    Kotani, Osamu; Suzuki, Tadaki; Yokoyama, Masaru; Iwata-Yoshikawa, Naoko; Nakajima, Noriko; Sato, Hironori; Hasegawa, Hideki; Taguchi, Fumihiro; Shimizu, Hiroyuki; Nagata, Noriyo

    2016-08-31

    Saffold virus (SAFV), a human cardiovirus, is occasionally detected in infants with neurological disorders, including meningitis and cerebellitis. We recently reported that SAFV type 3 isolates infect cerebellar glial cells, but not large neurons, in mice. However, the impact of this infection remained unclear. Here, we determined the neuropathogenesis of SAFV type 3 in the cerebella of neonatal ddY mice using SAFV passaged in the cerebellum of neonatal BALB/c mice. The virus titer in the cerebellum increased following inoculation of each of five passaged strains. The fifth passaged strain harbored amino acid substitutions in the VP2 (H160R and Q239R) and VP3 (K62M) capsid proteins. Molecular modeling of the capsid proteins suggested that the VP2-H160R and VP3-K62M mutations alter the structural dynamics of the receptor binding surface via formation of a novel hydrophobic interaction between the VP2 puff B and VP3 knob regions. When compared with the original strain, the passaged strain showed altered growth characteristics in human-derived astroglial cell lines and higher replication in the brains of neonatal mice. In addition, the passaged strain was more neurovirulent than the original strain, while both strains infected astroglial and neural progenitor cells in the mouse brain. Intracerebral inoculation of either the original or passaged strain affected brain Purkinje cell dendrites, and a high titer of the passaged strain induced cerebellar hypoplasia in neonatal mice. Thus, infection by mouse-passaged SAFV affected cerebellar development in neonatal mice. This animal model contributes to the understanding of the neuropathogenicity of SAFV infections in infants. Saffold virus (SAFV) is a candidate neuropathogenic agent in infants and children, but the neuropathogenicity of the virus has not been fully elucidated. Recently, we evaluated the pathogenicity of two clinical SAFV isolates in mice. Similar to other neurotropic picornaviruses, these isolates showed

  20. Co-Inoculation Effects of Thiobacillus thiooxidans Bacteria and Mycorrhiza (Glomus spp. on Maize Nutrition at Different Levels of Sulfur

    Directory of Open Access Journals (Sweden)

    A. Gholami

    2016-02-01

    . Inoculation withthesefungihas increased the yield of numerous field-grown crops. This study was aimed to evaluate the effects of thiobacillus bacteria and sulfur application on soil pH, and also their interactions with mycorrhizal fungi in order to improve nutrients uptake and grain yield of maize under alkaline soil condition. Materials and Methods: Treatments arranged as factorial experiment were based on RCBD with three replications. Treatments consisted of mycorrhizal inoculation: inoculated (m1 and non-inoculated (m0, thiobacillus in two levels of inoculated (t1 and non-inoculated (t0 and three levels of sulfur (S0: 0 kg.ha-1, S1: 250 kg.ha-1 and S2: 500 kg.ha-1. Four-row plots were prepared with row width and intra-row space of 60 and 20 cm, respectively. Seeds of maize (Zea Mays, Sc:647 were surface sterilized in a 10% (v/v solution of hydrogen peroxide for 10 min, were rinsed with sterile distilled water. Before sowing, 300 kg of urea per hectare were applied according to the results of soil analysis. In order to facilitate oxidation of sulfur to sulfate form, , S was applied and thoroughly mixed into top 30 cm of soil 30 days before sowing. One week before sowing, thiobacillus (Thiobacillus thiooxidans was inoculated. Inoculum of AM fungus Glomus intraradices, were added to soil just before planting at about 2 centimeters below seed sowing dept. To measure Arbuscular Mycorrhizal colonization, root plants collected one week before harvesting, cleared in 10% KOH at 80˚C for 2 h, and then acidified in 1% HCL for 60 min. Then the cleared roots were stained in a solution of Trypan blue. For nutrient analysis, the following procedure was applied. Zn, Fe, S, and P were determined by Inductively Coupled Plasma-atomic emission spectrometry apparatus. For this purpose, ash of seed samples was prepared at 500-550 degree of Celsius and then 5 ml of HCl 37% was added and with dionized water to reach to 50 ml. Kjeldahl method was used to determine nitrogen. Analysis of variance

  1. Effect of medical wet tissue on disinfection of object surface%医用消毒湿巾在医院内物体表面消毒效果观察

    Institute of Scientific and Technical Information of China (English)

    孙亚君

    2012-01-01

    OBJECTIVE To observe the effect of medical wet tissue on the diinfeciton of object surface. METHODS The suspension quantitative bactericidal te?L method was used for the identification of test disinfectant wipes for neutralizer, bactericidal disinfectant wipes for laboratory experiment was observed. Disinfection wet tissue of medical surface and chlorine-containing disinfectant were used respectively on the variety of materials and the tables of case HI > IV surfaces by using swab wipe sampling and bacteria count methods, the application effects of two disinfection methods in the hospital were detected. RESULTS The bacteria daub was taken on disinfection wet tissue directly, saturated for 2min, the logarithm value on average of killing Staphylococcusaureus and Escherich-ia coli were both > 3. 00; the disinfection wet tissue was applied on surface to wipe and disinfection for 2min, the logarithm value for killing natural bacteria on the surface was 1. 23; chlorine-containing disinfectant was applied on surface to wipe and disinfection for lOmin, the logarithm value for killing natural bacteria on the surface was 1. 18, the total bacteria colony on the objects surface after disinfection by the two sterilization methods were both less than 10 CFU/cm2. CONCLUSION The medical wet tissue is superior to chlorine-containing disinfectant in the disinfection of the object surface and is with simple operation, which is worthy of being promoted.%目的 观察医用消毒湿巾在医院内物体表面消毒效果.方法 采用悬液定量杀菌试验法,对消毒湿巾进行中和剂鉴定试验;对消毒湿巾进行实验室杀菌试验观察;分别用医用消毒湿巾和含氯消毒剂对医院内Ⅲ、Ⅳ类环境的各种物体表面及台面擦拭消毒,采用棉拭涂抹采样和细菌培养计数方法,对两种消毒方法在医院中的应用效果进行了检测.结果 在消毒湿巾上直接染菌,浸透作用2 min,对金黄色葡萄球菌和大肠埃希

  2. Immune Responses in Cattle Inoculated with Mycobacterium bovis, Mycobacterium tuberculosis, or Mycobacterium kansasii

    Science.gov (United States)

    Cattle were inoculated with Mycobacterium bovis, Mycobacterium tuberculosis, or Mycobacterium kansasii to compare antigen-specific immune responses to varied patterns of mycobacterial disease. Disease expression ranged from colonization with associated pathology (M. bovis), colonization without path...

  3. Field and greenhouse inoculation methods for assessment of sheath blight resistance in rice

    Directory of Open Access Journals (Sweden)

    Leila Garcês de Araújo

    2007-01-01

    Full Text Available Field and greenhouse inoculation methods were compared to determine the genetic variation for resistance toRhizoctonia solani in 38 somaclones of rice cultivar Metica-1. Rice plants in pots were inoculated with isolate 4F1 at the ageof sixty four days, with 2.0 g of the fungal culture, multiplied on rice grain and hull medium and placed on the soil surfacearound the plant. The differences among somaclones in relation to lesion height were significant and varied from 6.5 to 15.5cm. In the field trial of artificial inoculation with fungal culture, 52 days after planting, the lesion height varied from 6.2 to 17.7cm. The correlation between disease severity in the greenhouse and the field was positive and highly significant (r=0.44;P< 0.01, indicating the greenhouse inoculation as a safe method for screening germplasm for sheath blight resistance.

  4. Cytotoxic compounds against cancer cells from Bombyx mori inoculated with Cordyceps militaris.

    Science.gov (United States)

    Qiu, Weitao; Wu, Jing; Choi, Jae-Hoon; Hirai, Hirofumi; Nishida, Hiroshi; Kawagishi, Hirokazu

    2017-06-01

    Two compounds, 3'-deoxyinosine and cordycepin, were isolated from Bombyx mori inoculated with Cordyceps militaris. In the bioassay examining cytotoxicity against cancer cells, both compounds showed toxicity against A549, PANC-1, and MCF-7 cancer cells.

  5. In ovo inoculation of chicken embryos with probiotic bacteria and its effect on posthatch Salmonella susceptibility

    NARCIS (Netherlands)

    Oliveira, J.E. de; Hoeven-Hangoor, E. van der; Linde, I.B. van de; Montijn, R.C.; Vossen, J.M.B.M. van der

    2014-01-01

    The feasibility of establishing probiotic bacteria in the intestine of broiler chickens by in ovo inoculation was investigated, followed by verifying possible subsequent protection against Salmonella Enteriditis infection. In a first study, 7 commercially available probiotics were screened for

  6. The effect of an inoculant and enzymes on fermentation and nutritive value of sorghum straw silages.

    Science.gov (United States)

    Xing, L; Chen, L J; Han, L J

    2009-01-01

    The objectives of this study were to determine the effect of inoculant, enzymes and inoculant-enzymes mixture on fermentation quality, nutritive value, and microbial changes of sorghum straw silage. Sorghum straws were collected and treated with distilled water (control), inoculant, enzymes and inoculant+enzymes prior to ensiling. Three bag silos for each silage (denoted C, I, E and I+E, respectively) were opened after 3, 7, 11, 15, 30 and 60 days for chemical and microbial analyses. For all the silages, there was a rapid decline in pH during the first 3 days of ensiling. Relative to silage C, all the treatment (I, E and I+E) had higher (Psilages. Separate addition of two additives, especially for enzymes, can effectively (Psilage IVDMD and IVNDFD concentration. These results indicated that the addition of additives can improve the sorghum straw silage fermentation quality at different extent.

  7. Increased Biomass of Nursery-Grown Douglas-Fir Seedlings upon Inoculation with Diazotrophic Endophytic Consortia

    Directory of Open Access Journals (Sweden)

    Zareen Khan

    2015-10-01

    Full Text Available Douglas-fir (Pseudotsuga menziesii seedlings are periodically challenged by biotic and abiotic stresses. The ability of endophytes to colonize the interior of plants could confer benefits to host plants that may play an important role in plant adaptation to environmental changes. In this greenhouse study, nursery-grown Douglas-fir seedlings were inoculated with diazotrophic endophytes previously isolated from poplar and willow trees and grown for fifteen months in nutrient-poor conditions. Inoculated seedlings had significant increases in biomass (48%, root length (13% and shoot height (16% compared to the control seedlings. Characterization of these endophytes for symbiotic traits in addition to nitrogen fixation revealed that they can also solubilize phosphate and produce siderophores. Colonization was observed through fluorescent microscopy in seedlings inoculated with gfp- and mkate-tagged strains. Inoculation with beneficial endophytes could prove to be valuable for increasing the production of planting stocks in forest nurseries.

  8. In ovo inoculation of chicken embryos with probiotic bacteria and its effect on posthatch Salmonella susceptibility

    NARCIS (Netherlands)

    Oliveira, J.E. de; Hoeven-Hangoor, E. van der; Linde, I.B. van de; Montijn, R.C.; Vossen, J.M.B.M. van der

    2014-01-01

    The feasibility of establishing probiotic bacteria in the intestine of broiler chickens by in ovo inoculation was investigated, followed by verifying possible subsequent protection against Salmonella Enteriditis infection. In a first study, 7 commercially available probiotics were screened for compa

  9. Developing Soil Microbial Inoculants for Pest Management: Can One Have Too Much of a Good Thing?

    Science.gov (United States)

    Gadhave, Kiran R; Hourston, James E; Gange, Alan C

    2016-04-01

    Soil microbes present a novel a