Incubation of Aquilaria subintegra with Microbial Culture Supernatants Enhances Production of Volatile Compounds and Improves Quality of Agarwood Oil.

Science.gov (United States)

Monggoot, Sakon; Kulsing, Chadin; Wong, Yong Foo; Pripdeevech, Patcharee

2018-06-01

Incubation with microbial culture supernatants improved essential oil yield from Aquilaria subintegra woodchips. The harvested woodchips were incubated with de man, rogosa and sharpe (MRS) agar, yeast mold (YM) agar medium and six different microbial culture supernatants obtained from Lactobacillus bulgaricus , L. acidophilus , Streptococcus thermophilus , Lactococcus lactis , Saccharomyces carlsbergensis and S. cerevisiae prior to hydrodistillation. Incubation with lactic acid bacteria supernatants provided higher yield of agarwood oil (0.45% w/w) than that obtained from yeast (0.25% w/w), agar media (0.23% w/w) and water (0.22% w/w). The composition of agarwood oil from all media and microbial supernatant incubations was investigated by using gas chromatography-mass spectrometry. Overall, three major volatile profiles were obtained, which corresponded to water soaking (control), as well as, both YM and MRS media, lactic acid bacteria, and yeast supernatant incubations. Sesquiterpenes and their oxygenated derivatives were key components of agarwood oil. Fifty-two volatile components were tentatively identified in all samples. Beta-agarofuran, α-eudesmol, karanone, α-agarofuran and agarospirol were major components present in most of the incubated samples, while S. cerevisiae -incubated A. subintegra provided higher amount of phenyl acetaldehyde. Microbial culture supernatant incubation numerically provided the highest yield of agarwood oil compared to water soaking traditional method, possibly resulting from activity of extracellular enzymes produced by the microbes. Incubation of agarwood with lactic acid bacteria supernatant significantly enhanced oil yields without changing volatile profile/composition of agarwood essential oil, thus this is a promising method for future use.

Cervical Cancer Cell Supernatants Induce a Phenotypic Switch from U937-Derived Macrophage-Activated M1 State into M2-Like Suppressor Phenotype with Change in Toll-Like Receptor Profile

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Karina Sánchez-Reyes

2014-01-01

Full Text Available Cervical cancer (CC is the second most common cancer among women worldwide. Infection with human papillomavirus (HPV is the main risk factor for developing CC. Macrophages are important immune effector cells; they can be differentiated into two phenotypes, identified as M1 (classically activated and M2 (alternatively activated. Macrophage polarization exerts profound effects on the Toll-like receptor (TLR profile. In this study, we evaluated whether the supernatant of human CC cells HeLa, SiHa, and C-33A induces a shift of M1 macrophage toward M2 macrophage in U937-derived macrophages. Results. The results showed that soluble factors secreted by CC cells induce a change in the immunophenotype of macrophages from macrophage M1 into macrophage M2. U937-derived macrophages M1 released proinflammatory cytokines and nitric oxide; however, when these cells were treated with the supernatant of CC cell lines, we observed a turnover of M1 toward M2. These cells increased CD163 and IL-10 expression. The expression of TLR-3, -7, and -9 is increased when the macrophages were treated with the supernatant of CC cells. Conclusions. Our result strongly suggests that CC cells may, through the secretion of soluble factors, induce a change of immunophenotype M1 into M2 macrophages.

Microbiological water quality of Igapó Lake Londrina - PR and genotypic characterization of virulence factors associated with enteropathogenic Escherichia coli (EPEC and Shiga toxin-producing E. coli (STEC

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Paulo Alfonso Schuroff

2014-12-01

Full Text Available This work aimed at the detection and quantification of Total Coliforms and Escherichia coli in Igapó Lake, in order to evaluate the quality of these waters as proper or unfit for recreation, in addition to the genotypic characterization of virulence factors associated with EPEC and STEC by PCR. The study area was the Igapó Lake I, II, III and IV. Samples were collected monthly from March 2011 to February 2012. The technique used for the detection and quantification of Total Coliforms and E. coli was the Colilert chromogenic substrate. In the technique of PCR, the eae and bfp genes were tested to characterize the typical EPEC pathotype; stx1, stx2, eae and hlyA the STEC pathotype and the samples that presented only the eae gene were characterized as atypical EPEC. According to CONAMA Resolution 357/2005, it has been observed that only Igapó Lake III was rated inappropriate for primary contact recreation, while for secondary contact recreation, all lakes were considered appropriate. Moreover, a strong relationship between rainfall and E. coli indices in Igapó Lake can be observed, which in dry months the quantity drastically decreases, while in rainy months the opposite relationship was observed. Of the 97 strains of E. coli isolated, two had the eae gene (atypical EPEC. None of the isolates contained genes stx1, stx2, bfp and hlyA. Thus, we hope to educate the population and public agencies of the importance of microbiological monitoring of recreational waters to prevent outbreaks of waterborne infections.

Proteomics of apheresis platelet supernatants during routine storage: Gender-related differences.

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Dzieciatkowska, Monika; D'Alessandro, Angelo; Burke, Timothy A; Kelher, Marguerite R; Moore, Ernest E; Banerjee, Anirban; Silliman, Christopher C; West, Bernadette F; Hansen, Kirk C

2015-01-01

Proteomics has identified potential pathways involved in platelet storage lesions, which correlate with untoward effects in the recipient, including febrile non-haemolytic reactions. We hypothesize that an additional pathway involves protein mediators that accumulate in the platelet supernatants during routine storage in a donor gender-specific fashion. Apheresis platelet concentrates were collected from 5 healthy males and 5 females and routinely stored. The 14 most abundant plasma proteins were removed and the supernatant proteins from days 1 and 5 were analyzed via 1D-SDS-PAGE/nanoLC-MS/MS, before label-free quantitative proteomics analyses. Findings from a subset of 18 proteins were validated via LC-SRM analyses against stable isotope labeled standards. A total of 503 distinct proteins were detected in the platelet supernatants from the 4 sample groups: female or male donor platelets, either at storage day 1 or 5. Proteomics suggested a storage and gender-dependent impairment of blood coagulation mediators, pro-inflammatory complement components and cytokines, energy and redox metabolic enzymes. The supernatants from female donors demonstrated increased deregulation of structural proteins, extracellular matrix proteins and focal adhesion proteins, possibly indicating storage-dependent platelet activation. Routine storage of platelet concentrates induces changes in the supernatant proteome, which may have effects on the transfused patient, some of which are related to donor gender. The rationale behind this study is that protein components in platelet releasates have been increasingly observed to play a key role in adverse events and impaired homeostasis in transfused recipients. In this view, proteomics has recently emerged as a functional tool to address the issue of protein composition of platelet releasates from buffy coat-derived platelet concentrates in the blood bank. Despite early encouraging studies on buffy coat-derived platelet concentrates, platelet

Use of Bacillus thuringiensis supernatant from a fermentation process to improve bioremediation of chlorpyrifos in contaminated soils.

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Aceves-Diez, Angel E; Estrada-Castañeda, Kelly J; Castañeda-Sandoval, Laura M

2015-07-01

The aim of this research was to investigate the potential of a nutrient-rich organic waste, namely the cell-free supernatant of Bacillus thuringiensis (BtS) gathered from fermentation, as a biostimulating agent to improve and sustain microbial populations and their enzymatic activities, thereby assisting in the bioremediation of chlorpyrifos-contaminated soil at a high dose (70 mg kg(-1)). Experiments were performed for up to 80 d. Chlorpyrifos degradation and its major metabolic product, 3,5,6-trichloro-2-pyridinol (TCP), were quantified by high-performance liquid chromatography (HPLC); total microbial populations were enumerated by direct counts in specific medium; and fluorescein diacetate (FDA) hydrolysis was measured as an index of soil microbial activity. Throughout the experiment, there was higher chlorpyrifos degradation in soil supplemented with BtS (83.1%) as compared to non-supplemented soil. TCP formation and degradation occurred in all soils, but the greatest degradation (30.34%) was observed in soil supplemented with BtS. The total microbial populations were significantly improved by supplementation with BtS. The application of chlorpyrifos to soil inhibited the enzymatic activity; however, this negative effect was counteracted by BtS, inducing an increase of approximately 16% in FDA hydrolysis. These results demonstrate the potential of B. thuringiensis supernatant as a suitable biostimulation agent for enhancing chlorpyrifos and TCP biodegradation in chlorpyrifos-contaminated soils. Copyright © 2015 Elsevier Ltd. All rights reserved.

Captive wild birds as reservoirs of enteropathogenic E. coli (EPEC) and Shiga-toxin producing E. coli (STEC).

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Sanches, Lilian Aparecida; Gomes, Marcelo da Silva; Teixeira, Rodrigo Hidalgo Friciello; Cunha, Marcos Paulo Vieira; Oliveira, Maria Gabriela Xavier de; Vieira, Mônica Aparecida Midolli; Gomes, Tânia Aparecida Tardelli; Knobl, Terezinha

Psittacine birds have been identified as reservoirs of diarrheagenic Escherichia coli, a subset of pathogens associated with mortality of children in tropical countries. The role of other orders of birds as source of infection is unclear. The aim of this study was to perform the molecular diagnosis of infection with diarrheagenic E. coli in 10 different orders of captive wild birds in the state of São Paulo, Brazil. Fecal samples were analyzed from 516 birds belonging to 10 orders: Accipitriformes, Anseriformes, Columbiformes, Falconiformes, Galliformes, Passeriformes, Pelecaniformes, Piciformes, Psittaciformes and Strigiformes. After isolation, 401 E. coli strains were subjected to multiplex PCR system with amplification of genes eae and bfp (EPEC), stx1 and stx2 for STEC. The results of these tests revealed 23/401 (5.74%) positive strains for eae gene, 16/401 positive strains for the bfp gene (3.99%) and 3/401 positive for stx2 gene (0.75%) distributed among the orders of Psittaciformes, Strigiformes and Columbiformes. None of strains were positive for stx1 gene. These data reveal the infection by STEC, typical and atypical EPEC in captive birds. The frequency of these pathotypes is low and restricted to few orders, but the data suggest the potential public health risk that these birds represent as reservoirs of diarrheagenic E. coli. Copyright © 2017 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

Cell mediated lympholysis: CML. A microplate technique requiring few target cells and employing a new method of supernatant collection

International Nuclear Information System (INIS)

Hirschberg, A.; Thorsby, E.

1977-01-01

A micromethod for the 51 Cr release assay is described. Allogeneically induced cytotoxic lymphocytes are generated in mixed lymphocyte microcultures in the wells of microplates. Their cytotoxic capacity is assayed by adding 51 Cr-labelled PHA derived lymphoblasts directly into the microcultures with no pooling or transfer of the cytotoxic effector cells being required. The 51 Cr isotope released into the cell supernatants is collected by inserting a cellulose acetate absorption cartridge into each well. A glass fiber filter attached to the cartridge effectively separates the supernatant from the cellular elemets. This system allows the simultaneous collection of the supernatant from 96 wells, and can be used with either adherent or non-adherent target cells

Measuring medium-induced gluons via jet grooming

Science.gov (United States)

Tywoniuk, Konrad; Mehtar-Tani, Yacine

2017-11-01

Jet substructure observables and applications of jet grooming techniques in heavy-ion collisions are still in its infancy and provide new alleys for studying medium modifications of perturbative degrees of freedom. We note that these measurements, given the right transverse momentum range, can be uniquely sensitive to rare medium-induced emissions inside of the jet cone. This corresponds to an infrared enhancement that would, for instance, affect the distribution of the groomed momentum-sharing variable zg measured using the SoftDrop procedure.

Endocytosis-inducer adhesins produced by enteropathogenic serogroups of Escherichia coli participate on bacterial attachment to infant enterocytes

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João Ramos Costa Andrade

1987-03-01

Full Text Available Enteropathogenic E. coli (EPEC infection of Hep-2 cells preoceeds through bacterial attachment to cell surface and internalization of adhered bacteria. EPEC attachment is a prerequisite for cell infection and is mediated by adhesins that recognize carbohydrate-containing receptors on cell membrane. Such endocytosis-inducer adhesins (EIA also promote EPEC binding to infant enterocytes, suggesting that EIA may have an important role on EPEC gastroenteritis.A infecção de células Hep-2 por E. coli enteropatogênicas (ECEP implica na aderência bacteriana e posterior interiorização dos microrganismos aderidos por um mecanismo de endocitose. A aderência das ECEP é pré-requisito para a infecção e é mediada por adesinas que reconhecem receptores inibidos por certas oses na membrana celular. Tais "adesinas indutoras da endocitose" (AIE também promovem a ligação bacteriana a enterócitos obtidos do intestino delgado de lactente, sugerindo que as AIE possam desempenhar algum papel nas diarréias causadas por ECEP.

Diarréia aguda grave associada à Escherichia coli enteropatogênica clássica (EPEC: características clínicas e perdas fecais em lactentes hospitalizados Severe acute diarrhea associated to classic enteropathogenic by clinical features and fecal losses in hospitalized infants

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C.A.G. Oliva

1997-12-01

estudados. A duração da diarréia mostrou curso agudo (menor ou igual a 14 dias na maioria das crianças estudadas, reafirmando a evolução potencialmente autolimitada da diarréia aguda. Sete (18,4% pacientes, entretanto, desenvolveram quadro de diarréia persistente, indicando que tal evolução é esperada em uma parcela razoável de lactentes jovens hospitalizados com tal enfermidade. Os volumes médios de ingestão das fórmulas alimentares encontrados mostram o impacto da diarréia aguda na diminuição da ingestão alimentar. As médias observadas nas três diferentes fórmulas alimentares ficaram abaixo da ingestão calórica sugerida como padrão pela FAO/WHO (100cal/kg/dia. Não houve diferenças entre os volumes de aceitação segundo os tipos de fórmulas alimentares utilizadas.Enteropathogenic Escherichia coli (EPEC strains are the most prevalent enteropathogenic agents isolated in the stools of hospitalized infants with severe acute diarrhea in São Paulo. These microorganisms induce a severe intestinal secretion of fluids and electrolytes that can cause dehydration leading to hospital admission in the majority of the cases. OBJECTIVES. This investigation aims at the following objectives: 1 to study the clinical features of acute diarrhea in male infants who were hospitalized owing to EPEC infection; 2 to determine the fecal fluid losses, formula intake and variation of the body weight during the evolution of the disease. PATIENTS AND METHODS. Thirty eight male infants with acute diarrhea due to EPEC strains were studied. The clinical and epidemiological features of the patients were recorded and the different EPEC serogroups were identified. The infants were kept in a metabolic bed in order to allow the collection of stools and urine separately during the whole period of the disease, and daily metabolic balances were also obtained. The mean duration of the metabolic study was 5.8 days, and during this period of time 220 daily metabolic balances were

Half-liter supernatant sampler system engineering work plan

International Nuclear Information System (INIS)

Ritter, G.A.

1995-01-01

The Tank Waste Remediation System (TWRS) pretreatment facility project W-236B, known as the Initial Pretreatment Module (IPM), requires samples of supernatants and sludges from 200 Area tank farms for planned hot testing work in support of IPM design. The IPM project has proposed the development of several new sampler systems. These systems include a 0.5-l supernatant sampler, 3-l and 25-l supernatant and sludge samplers, and a 4,000-l sampler system. The 0.5-l sampler will support IPM sampling needs in the 1 to 3 l range starting in late fiscal year 1995. This sampler is intended to be used in conjunction with the existing 100 ml bottle-on-a-string. The 3-l and 25-l systems will be based on the Savannah River Site's sampler system and will support IPM sampling needs in the 3 to 100 liter range. Most of the hot testing required for design of the IPM must be accomplished in the next 3 years. This work plan defines the tasks associated with the development of a 0.5-l sampler system. This system will be referred to as the Half-Liter Supernatant Sampler System (HLSSS). Specifically, this work plan will define the scope of work, identify organizational responsibilities, identify major technical requirements, describe configuration control and verification requirements, and provide estimated costs and schedule. The sampler system will be fully operational, including trained staff and operating procedures, upon completion of this task

[Effects of combined application of culture supernatant of human umbilical cord mesenchymal stem cells and ciprofloxacin on Staphylococcus aureus in vitro].

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Zhou, B; Tu, H L; Ba, T; Wang, L F; Wang, S J; Nie, S Y

2017-06-20

Objective: To explore the effects of combined application of culture supernatant of human umbilical cord mesenchymal stem cells (hUCMSCs) and ciprofloxacin on Staphylococcus aureus (SA) in vitro. Methods: hUCMSCs were isolated from umbilical cord tissue of full-term healthy fetus after cesarean section and cultured. Cells in the third passage were used in the experiments after identification. SA strains isolated from wounds of burn patients in our burn wards were used in the experiments. Cells were divided into 0, 10, 100, and 1 000 ng/mL lipopolysaccharide (LPS) groups according to the random number table (the same dividing method below). Cells were cultured with culture medium of mesenchymal stem cells (MSCs) after being treated with medium containing the corresponding mass concentrations of LPS for 12 h. At post culture hour (PCH) 6, 12, and 24, 6 wells of culture supernatant of cells in each group were obtained to measure the content of LL-37 with enzyme-linked immunosorbent assay. Ninety blood agar plates were divided into ciprofloxacin control group (CC), ciprofloxacin+ supernatant group (CS), and ciprofloxacin+ supernatant+ LL-37 antibody group (CSL), with 30 blood agar plates in each group. Blood agar plates in group CC were coated with 1.5×10(8) colony forming unit (CFU)/mL bacteria solution prepared with normal saline. Blood agar plates in group CS were coated with 1.5×10(8) CFU/mL bacteria solution prepared with normal saline and culture supernatant of hUCMSCs (cultured by culture medium of MSCs, the same below) in double volume of normal saline. Blood agar plates in group CSL were coated with 1.5×10(8) CFU/mL bacteria solution prepared with normal saline, culture supernatant of hUCMSCs in double volume of normal saline, and 2.6 μL LL-37 antibody in the concentration of 2 μg/mL. At PCH 12, 24, and 48, 10 blood agar plates of each group were harvested to observe the distribution of SA colony on blood agar plate and to measure the diameter of

Potential mechanisms behind contrast medium-induced nephropathy

African Journals Online (AJOL)

How contrast medium-induced nephropathy (CIN) comes about is poorly understood, although CIN is a common cause of acute renal failure. Hitherto, the various studies performed have led to different interpretations and partially contradictory conclusions. This article aimed to review the mechanisms underlying CIN and to ...

Identificación de péptidos bloqueantes del sistema de secreción de tipo III y de la adherencia a epitelios de Escherichia coli enterohemorrágico (EHEC) y Escherichia coli enteropatógeno (EPEC)

OpenAIRE

Larzábal, Mariano

2010-01-01

Escherichia coli Enteropatógeno (EPEC) y Escherichia coli Enterohemorrágico (EHEC) se encuentran asociadas con diarrea en seres humanos. EPEC es una de las principales causas de diarrea infantil en países desarrollados, mientras que EHEC es responsable de enfermedades cuyo espectro clínico incluye diarrea, colitis hemorrágica y síndrome urémico hemolítico (SUH), que es la principal causa de insuficiencia renal aguda de niños menores de 5 años en la Argentina. La expresión de la toxina Shiga (...

Oral administration of a select mixture of Bacillus probiotics generates Tr1 cells in weaned F4ab/acR- pigs challenged with an F4+ ETEC/VTEC/EPEC strain.

Science.gov (United States)

Zhou, Dong; Zhu, Yao-Hong; Zhang, Wei; Wang, Meng-Ling; Fan, Wen-Yi; Song, Dan; Yang, Gui-Yan; Jensen, Bent Borg; Wang, Jiu-Feng

2015-09-17

Although breeding of F4 receptor - negative (F4R(-)) pigs may prevent post-weaning diarrhea, the underlying immunity is poorly understood. Here, various doses of a Bacillus licheniformis and Bacillus subtilis mixture (BLS-mix) were orally administered to F4ab/acR(-) pigs for 1 week before F4 (K88) - positive ETEC/VTEC/EPEC challenge. Administration of BLS-mix increased the percentage of Foxp3(-)IL-10(+) T cells but not of Foxp3(+)IL-10(+) regulatory T (Treg) cells among peripheral blood CD4(+) T cells. A low dose of BLS-mix feeding resulted in increased the expression of IL-6, TNF-α, IL-10, and the transcription factors Foxp3 and T-bet mRNAs in the jejunum. Administration of either a low or high dose BLS-mix also led to an increase in the percentage of CD4(+)Foxp3(+) Treg cells among intraepithelial lymphocytes and CD4(+)IL-10(+) T cells in the small intestinal Peyer's patches and the lamina propria of F4ab/acR(-) pigs following F4(+) ETEC/VTEC/EPEC challenge. The increased number of IL-10-producing CD4(+) T cells was attributed to an increase in the proportion of Foxp3(-)IL-10(+) Treg cells rather than Foxp3(+)IL-10(+) Treg cells. Our data indicate that oral administration of BLS-mix to newly weaned F4ab/acR(-) pigs ameliorates enteritis in an F4(+) ETEC/VTEC/EPEC model; however, induction of IL-10-producing Foxp3(-) Treg cells by BLS-mix administration cannot account for the protection of newly weaned F4ab/acR(-) pigs from F4(+) ETEC/VTEC/EPEC infection, and that excessive generation of CD4(+)IL-10(+) T cells following consumption of BLS-mix during episodes of intestinal inflammation that is caused by enteric pathogens might prohibit clearance of the pathogen. Select probiotic mixtures may allow for tailoring strategies to prevent infectious diseases.

Endogenous CO2 may inhibit bacterial growth and induce virulence gene expression in enteropathogenic Escherichia coli.

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Martínez, Haydee; Buhse, Thomas; Rivera, Marco; Parmananda, P; Ayala, Guadalupe; Sánchez, Joaquín

2012-07-01

Analysis of the growth kinetics of enteropathogenic Escherichia coli (EPEC) revealed that growth was directly proportional to the ratio between the exposed surface area and the liquid culture volume (SA/V). It was hypothesized that this bacterial behavior was caused by the accumulation of an endogenous volatile growth inhibitor metabolite whose escape from the medium directly depended on the SA/V. The results of this work support the theory that an inhibitor is produced and indicate that it is CO(2). We also report that concomitant to the accumulation of CO(2), there is secretion of the virulence-related EspB and EspC proteins from EPEC. We therefore postulate that endogenous CO(2) may have an effect on both bacterial growth and virulence. Copyright © 2012 Elsevier Ltd. All rights reserved.

Growth Inhibition of Cronobacter sakazakii in Experimentally Contaminated Powdered Infant Formula by Kefir Supernatant.

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Kim, Dong-Hyeon; Chon, Jung-Whan; Kang, Il-Byeong; Kim, Hyunsook; Kim, Hong-Seok; Song, Kwang-Young; Seo, Kun-Ho

2015-09-01

Kefir is a type of fermented milk containing lactic and acetic acid bacteria and yeast. In this study, we evaluated the antimicrobial activity of kefir supernatant against Cronobacter sakazakii in powdered infant formula (PIF). In a spot-on-lawn test, the growth of 20 C. sakazakii strains, including 10 clinical and 10 food isolates, was completely inhibited in the presence of kefir supernatant. Significant differences in the diameters of inhibition zones were observed upon treatment with kefir compared with the results for Lactobacillus kefiri and Candida kefyr culture supernatants or solutions of lactic and acetic acid and ethyl alcohol in the agar well diffusion test (P < 0.05). The addition of 100 μl of kefir supernatant to 1 ml of nutrient broth completely inhibited the growth of C. sakazakii, as evaluated by spectrophotometry. The antimicrobial activity of kefir supernatant in experimentally contaminated PIF was also tested; we found no viable C. sakazakii cells remaining in PIF rehydrated with 30% kefir supernatant solution for 1 h, demonstrating that the antimicrobial activity of kefir supernatant against C. sakazakii could be applied in real food samples.

Residual urinary extracellular vesicles in ultracentrifugation supernatants after hydrostatic filtration dialysis enrichment.

Science.gov (United States)

Musante, Luca; Tataruch-Weinert, Dorota; Kerjaschki, Dontscho; Henry, Michael; Meleady, Paula; Holthofer, Harry

2017-01-01

Urinary extracellular vesicles (UEVs) appear an ideal source of biomarkers for kidney and urogenital diseases. The majority of protocols designed for their isolation are based on differential centrifugation steps. However, little is still known of the type and amount of vesicles left in the supernatant. Here we used an isolation protocol for UEVs which uses hydrostatic filtration dialysis as first pre-enrichment step, followed by differential centrifugation. Transmission electron microscopy (TEM), mass spectrometry (MS), western blot, ELISA assays and tuneable resistive pulse sensing (TRPS) were used to characterise and quantify UEVs in the ultracentrifugation supernatant. TEM showed the presence of a variety of small size vesicles in the supernatant while protein identification by MS matched accurately with the protein list available in Vesiclepedia. Screening and relative quantification for specific vesicle markers showed that the supernatant was preferentially positive for CD9 and TSG101. ELISA tests for quantification of exosome revealed that 14%, was left in the supernatant with a particle diameter of 110 nm and concentration of 1.54 × 10 10 /ml. Here we show a comprehensive characterisation of exosomes and other small size urinary vesicles which the conventional differential centrifugation protocol may lose.

Electromagnetically Induced Transparency In Rydberg Atomic Medium

Science.gov (United States)

Deng, Li; Cong, Lu; Chen, Ai-Xi

2018-03-01

Due to possessing big principal quantum number, Rydberg atom has some unique properties, for example: its radiative lifetime is long, dipole moment is large, and interaction between atoms is strong and so on. These properties make one pay attention to Rydberg atoms. In this paper we investigate the effects of Rydberg dipole-dipole interactions on electromagnetically induced transparency (EIT) schemes and group velocity in three-level systems of ladder type, which provides theoretical foundation for exploring the linear and nonlinear characteristics of light in a Rydberg electromagnetically-induced-transparency medium.

Radiation-induced organogenesis: effects of irradiated medium and its components on tobacco tissue culture

International Nuclear Information System (INIS)

Degani, N.

1975-01-01

Gamma irradiated medium induces the formation of buds in non-irradiated dark growth tobacco callus (Nicotiana tabacum Var. Wisconsin No.38). Experiments were conducted to determine the component(s) of the medium that is effective in this radiation-induced organogenesis. Fraction of medium were irradiated singly and in combination, then combined with non-irradiated fractions to form the complete growth medium. The results showed that irradiated indoleacetic acid (IAA) was not the effective component in the induction of organogensis. Omission of IAA from the medium resulted in the formation of buds, as expected. Irradiated myo-inositol induced organogenesis more consistently than the other irradiated components. The age of the inoculum tissue and its passage number from the tobacco stem affected the potency of the tobacco callus to organise. (author)

Fermentation supernatants of Lactobacillus delbrueckii inhibit growth of human colon cancer cells and induce apoptosis through a caspase 3-dependent pathway.

Science.gov (United States)

Wan, Ying; Xin, Yi; Zhang, Cuili; Wu, Dachang; Ding, Dapeng; Tang, Li; Owusu, Lawrence; Bai, Jing; Li, Weiling

2014-05-01

Probiotic bacteria are known to exert a wide range of beneficial effects on their animal hosts. Therefore, the present study explored the effect of the supernatants obtained from Lactobacillus delbrueckii fermentation (LBF) on colon cancer. The results indicated that the proliferation of LBF solution-treated colon cancer SW620 cells was arrested and accumulated in the G1 phase in a concentration-dependent manner. The LBF solution efficiently induced apoptosis through the intrinsic caspase 3-depedent pathway, with a corresponding decreased expression of Bcl-2. The activity of matrix metalloproteinase 9, which is associated with the invasion of colon cancer cells, was also decreased in the LBF-treated cells. In conclusion, the results demonstrate the antitumor effect of LBF in vitro and may contribute to the development of novel therapies for the treatment of colon cancer.

Light-induced retinal injury enhanced neurotrophins secretion and neurotrophic effect of mesenchymal stem cells in vitro

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Wei Xu

2013-04-01

Full Text Available PURPOSE: To investigate neurotrophins expression and neurotrophic effect change in mesenchymal stem cells (MSCs under different types of stimulation. METHODS: Rats were exposed in 10,000 lux white light to develop light-induced retinal injury. Supernatants of homogenized retina (SHR, either from normal or light-injured retina, were used to stimulate MSCs. Quantitative real time for polymerase chain reaction (RT-PCR and enzyme-linked immunosorbent assay (ELISA were conducted for analysis the expression change in basic fibroblast growth factor (bFGF, brain-derived neurotrophic factor (BDNF and ciliary neurotrophic factor (CNTF in MSCs after stimulation. Conditioned medium from SHR-stimulated MSCs and control MSCs were collected for evaluation their effect on retinal explants. RESULTS: Supernatants of homogenized retina from light-injured rats significantly promoted neurotrophins secretion from MSCs (p<0.01. Conditioned medium from mesenchymal stem cells stimulated by light-injured SHR significantly reduced DNA fragmentation (p<0.01, up-regulated bcl-2 (p<0.01 and down-regulated bax (p<0.01 in retinal explants, displaying enhanced protective effect. CONCLUSIONS: Light-induced retinal injury is able to enhance neurotrophins secretion from mesenchymal stem cells and promote the neurotrophic effect of mesenchymal stem cells.

Measuring and slowing decoherence in Electromagnetically induced transparency medium

International Nuclear Information System (INIS)

Shuker, M.; Firstenberg, O.; Sagi, Y.; Ben-Kish, A.; Fisher, A.; Ron, A.; Davidson, N.

2005-01-01

Full Text:Electromagnetically induced transparency is a unique light-matter interaction that exhibits extremely narrow-band spectroscopic features along with low absorption. Recent interest in this phenomenon is driven by its possible applications in quantum information (slow light, storage of light), atomic clocks and precise magnetometers. The Electromagnetically induced transparency phenomenon takes place when an atomic ensemble is driven to a coherent superposition of its ground state sub-levels by two phase-coherent radiation fields. A key parameter of the Electromagnetically induced transparency medium, that limits its applicability, is the coherence lifetime of this superposition (decoherence rate). We have developed a simple technique to measure decay rates within the ground state of an atomic ensemble, and specifically the decoherence rate of the Electromagnetically induced transparency coherent superposition. Detailed measurements were performed in a Rubidium vapor cell at 60 - 80 with 30 Torr of Neon buffer gas. We have found that the Electromagnetically induced transparency decoherence is dominated by spin-exchange collisions between Rubidium atoms. We discuss the sensitivity of various quantum states of the atomic ensemble to spin exchange decoherence, and find a set of quantum states that minimize this effect. Finally, we demonstrate a unique quantum state which is both insensitive to spin exchange decoherence and constitutes an Electromagnetically induced transparency state of the medium

Gastric cancer cell supernatant causes apoptosis and fibrosis in the peritoneal tissues and results in an environment favorable to peritoneal metastases, in vitro and in vivo

Directory of Open Access Journals (Sweden)

Na Di

2012-04-01

Full Text Available Abstract Background In this study, we examined effects of soluble factors released by gastric cancer cells on peritoneal mesothelial cells in vitro and in vivo. Methods HMrSV5, a human peritoneal mesothelial cell line, was incubated with supernatants from gastric cancer cells. Morphological changes of HMrSV5 cells were observed. Apoptosis of HMrSV5 cells was observed under a transmission electron microscope and quantitatively determined by MTT assay and flow cytometry. Expressions of apoptosis-related proteins (caspase-3, caspase-8, Bax, bcl-2 were immunochemically evaluated. Results Conspicuous morphological changes indicating apoptosis were observed in HMrSV5 cells 24 h after treatment with the supernatants of gastric cancer cells. In vivo, peritoneal tissues treated with gastric cancer cell supernatant were substantially thickened and contained extensive fibrosis. Conclusions These findings demonstrate that supernatants of gastric cancer cells can induce apoptosis and fibrosis in HMrSV5 human peritoneal mesothelial cells through supernatants in the early peritoneal metastasis, in a time-dependent manner, and indicate that soluble factors in the peritoneal cavity affect the morphology and function of mesothelial cells so that the resulting environment can become favorable to peritoneal metastases.

Test procedures and instructions for Hanford tank waste supernatant cesium removal

Energy Technology Data Exchange (ETDEWEB)

Hendrickson, D.W., Westinghouse Hanford

1996-05-31

This document provides specific test procedures and instructions to implement the test plan for the preparation and conduct of a cesium removal test using Hanford Double-Shell Slurry Feed supernatant liquor from tank 251-AW-101 in a bench-scale column.Cesium sorbents to be tested include resorcinol-formaldehyde resin and crystalline silicotitanate. The test plan for which this provides instructions is WHC-SD-RE-TP-022, Hanford Tank Waste Supernatant Cesium Removal Test Plan.

Diagnosing feline infectious peritonitis using the Sysmex XT-2000iV based on frozen supernatants from cavitary effusions.

Science.gov (United States)

Stranieri, Angelica; Paltrinieri, Saverio; Giordano, Alessia

2017-05-01

The delta total nucleated cells (ΔTNC) measurement with the Sysmex XT-2000iV (Sysmex Europe, Norderstedt, Germany) has high diagnostic accuracy on effusions in feline infectious peritonitis (FIP) cases, but the test can be performed only on fresh samples. We evaluated whether supernatants from effusions retain the ability to induce cell clumping and assessed the diagnostic accuracy of this modified ΔTNC method. Effusions were collected from FIP cats ( n = 19) and from cats with other diseases ( n = 15). ΔTNC was measured on fresh samples and on frozen-thawed supernatants after the addition of feline blood at 1:10 dilution. Diagnostic accuracy was assessed at the cutoffs of suggestive of FIP (ΔTNC = 1.7) and consistent with FIP (ΔTNC = 3.4). The influence of the protein content, number of added cells, and magnitude of dilution were also investigated. Specificity and positive predictive value were 100% for both the methods. Sensitivity and negative predictive value were higher for the modified ΔTNC (84.2% and 83.3%, respectively, at the cutoff of 1.7; 78.9% and 78.9%, respectively, at the cutoff of 3.4) than for the ΔTNC on fresh samples (78.6% and 81.3%, respectively, at the cutoff of 1.7; 57.1% and 68.4%, respectively, at the cutoff of 3.4). Protein content, total cell count of the added blood, and magnitude of dilutions did not influence the results. Supernatants of frozen effusions from FIP cats retain the ability to induce cell clumping, thus the modified ΔTNC measurement is a reliable tool to diagnose FIP on samples that cannot be analyzed immediately.

Fate of cyanobacteria in drinking water treatment plant lagoon supernatant and sludge

International Nuclear Information System (INIS)

Pestana, Carlos J.; Reeve, Petra J.; Sawade, Emma; Voldoire, Camille F.; Newton, Kelly; Praptiwi, Radisti; Collingnon, Lea; Dreyfus, Jennifer; Hobson, Peter; Gaget, Virginie; Newcombe, Gayle

2016-01-01

In conventional water treatment processes, where the coagulation and flocculation steps are designed to remove particles from drinking water, cyanobacteria are also concentrated into the resultant sludge. As a consequence, cyanobacteria-laden sludge can act as a reservoir for metabolites such as taste and odour compounds and cyanotoxins. This can pose a significant risk to water quality where supernatant from the sludge treatment facility is returned to the inlet to the plant. In this study the complex processes that can take place in a sludge treatment lagoon were investigated. It was shown that cyanobacteria can proliferate in the conditions manifest in a sludge treatment lagoon, and that cyanobacteria can survive and produce metabolites for at least 10 days in sludge. The major processes of metabolite release and degradation are very dependent on the physical, chemical and biological environment in the sludge treatment facility and it was not possible to accurately model the net effect. For the first time evidence is provided to suggest that there is a greater risk associated with recycling sludge supernatant than can be estimated from the raw water quality, as metabolite concentrations increased by up to 500% over several days after coagulation, attributed to increased metabolite production and/or cell proliferation in the sludge. - Highlights: • Cyanobacteria in water treatment sludge significantly impact supernatant quality • Cyanobacteria can survive, and thrive, in sludge lagoon supernatant and in treatment sludge • Metabolite concentrations in cyanobacteria in sludge can increase up to 500% • The risk associated with supernatant recycling was assessed relative to available treatment barriers

Fate of cyanobacteria in drinking water treatment plant lagoon supernatant and sludge

Energy Technology Data Exchange (ETDEWEB)

Pestana, Carlos J.; Reeve, Petra J.; Sawade, Emma [Australian Water Quality Centre, South Australian Water Corporation, Adelaide, SA 5000 (Australia); Voldoire, Camille F. [Australian Water Quality Centre, South Australian Water Corporation, Adelaide, SA 5000 (Australia); École Européenne de Chimie, Polymères et Matériaux (ECPM), Strasbourg 67087 (France); Newton, Kelly; Praptiwi, Radisti [Australian Water Quality Centre, South Australian Water Corporation, Adelaide, SA 5000 (Australia); Collingnon, Lea [Australian Water Quality Centre, South Australian Water Corporation, Adelaide, SA 5000 (Australia); École Européenne de Chimie, Polymères et Matériaux (ECPM), Strasbourg 67087 (France); Dreyfus, Jennifer [Allwater, Adelaide Services Alliance, Wakefield St, Adelaide, SA 5001 (Australia); Hobson, Peter [Australian Water Quality Centre, South Australian Water Corporation, Adelaide, SA 5000 (Australia); Gaget, Virginie [University of Adelaide, Ecology and Environmental Sciences, School of Biological Sciences, Adelaide, SA 5005 (Australia); Newcombe, Gayle, E-mail: gayle.newcombe@sawater.com.au [Australian Water Quality Centre, South Australian Water Corporation, Adelaide, SA 5000 (Australia)

2016-09-15

In conventional water treatment processes, where the coagulation and flocculation steps are designed to remove particles from drinking water, cyanobacteria are also concentrated into the resultant sludge. As a consequence, cyanobacteria-laden sludge can act as a reservoir for metabolites such as taste and odour compounds and cyanotoxins. This can pose a significant risk to water quality where supernatant from the sludge treatment facility is returned to the inlet to the plant. In this study the complex processes that can take place in a sludge treatment lagoon were investigated. It was shown that cyanobacteria can proliferate in the conditions manifest in a sludge treatment lagoon, and that cyanobacteria can survive and produce metabolites for at least 10 days in sludge. The major processes of metabolite release and degradation are very dependent on the physical, chemical and biological environment in the sludge treatment facility and it was not possible to accurately model the net effect. For the first time evidence is provided to suggest that there is a greater risk associated with recycling sludge supernatant than can be estimated from the raw water quality, as metabolite concentrations increased by up to 500% over several days after coagulation, attributed to increased metabolite production and/or cell proliferation in the sludge. - Highlights: • Cyanobacteria in water treatment sludge significantly impact supernatant quality • Cyanobacteria can survive, and thrive, in sludge lagoon supernatant and in treatment sludge • Metabolite concentrations in cyanobacteria in sludge can increase up to 500% • The risk associated with supernatant recycling was assessed relative to available treatment barriers.

Development of a rapid agglutination latex test for diagnosis of enteropathogenic and enterohemorrhagic Escherichia coli infection in developing world: defining the biomarker, antibody and method.

Directory of Open Access Journals (Sweden)

Letícia B Rocha

2014-09-01

Full Text Available Enteropathogenic and enterohemorrhagic Escherichia coli (EPEC/EHEC are human intestinal pathogens responsible for diarrhea in both developing and industrialized countries. In research laboratories, EPEC and EHEC are defined on the basis of their pathogenic features; nevertheless, their identification in routine laboratories is expensive and laborious. Therefore, the aim of the present work was to develop a rapid and simple assay for EPEC/EHEC detection. Accordingly, the EPEC/EHEC-secreted proteins EspA and EspB were chosen as target antigens.First, we investigated the ideal conditions for EspA/EspB production/secretion by ELISA in a collection of EPEC/EHEC strains after cultivating bacterial isolates in Dulbecco's modified Eagle's medium (DMEM or DMEM containing 1% tryptone or HEp-2 cells-preconditioned DMEM, employing either anti-EspA/anti-EspB polyclonal or monoclonal antibodies developed and characterized herein. Subsequently, a rapid agglutination latex test (RALT was developed and tested with the same collection of bacterial isolates.EspB was defined as a biomarker and its corresponding monoclonal antibody as the tool for EPEC/EHEC diagnosis; the production of EspB was better in DMEM medium. RALT assay has the sensitivity and specificity required for high-impact diagnosis of neglected diseases in the developing world.RALT assay described herein can be considered an alternative assay for diarrhea diagnosis in low-income countries since it achieved 97% sensitivity, 98% specificity and 97% efficiency.

Fluorescence Spectrum and Decay Measurement for Hsil VS Normal Cytology Differentiation in Liquid Pap Smear Supernatant

Science.gov (United States)

Vaitkuviene, A.; Gegzna, V.; Juodkazis, S.; Jursenas, S.; Miasojedovas, S.; Kurtinaitiene, R.; Rimiene, J.; Vaitkus, J.

2009-06-01

Cervical smear material contains endo and exocervical cells, mucus and inflammative, immune cells in cases of pathology. Just not destroyed keratinocytes lay on the glass for microscopy. Liquid cytology supernatant apart other diagnostics could be used for photodiagnostic. The spectroscopic parameters suitable for Normal and HSIL cytology groups supernatant differentiation are demonstrated. The dried liquid PAP supernatant fractions—sediment and liquid were investigated. Excitation and emission matrices (EEM), supernatant fluorescence decay measured under 280 nm diode short pulse excitation and fluorescence spectroscopy by excitation with 355 nm laser light were analyzed. The differences between Normal and HSIL groups were statistically proven in the certain spectral regions. Fluorescence decay peculiarities show spectral regions consisting of few fluorophores. Obtained results on fluorescence differences in Normal and HSIL groups' supernatant shows the potency of photodiagnosis application in cervical screening.

Assessment of chemical composition and microbiological properties of titanium nickelide supernatant

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Uruzbaev R.M.

2018-03-01

Full Text Available According to the World Health Organization, in the structure of internal injuries, burns occur in 2.1 per 1000 adults. In practice, all victims after stabilization of the condition are treated surgically with the autodermoplasty, which in the late period is accompanied by a rough scarring, sometimes with the deformation of the damaged area. In addition, infectious complications worsen and aggravate the wound healing process, and also significantly prolong the epithelialization time. A wound is usually infected by opportunistic pathogenic microflora resistant to various groups of antibiotics and antiseptics, by which the affected areas are treated. Modern researches are aimed at finding ways to accelerate regenerative processes of the affected area, to avoid surgical treatment and prevent infection contamination. The object of this study was titanium nickelide supernatant. Currently, there are no published data on the chemical composition of the supernatant, as well as its microbiological safety. This alloy is actively and successfully used in traumatology, dentistry and other fields of medicine. The supernatant was prepared under sterile conditions at a rate of 10 g of titanium nickelide powder per 1 liter of distilled water. In the course of the studies it was proved that nickel and titanium ions are present in the solution, in addition, its bactericidal and bacteriostatic properties are reflected. Thus, the supernatant of nitinol can be used in biological environment and is safe for them.

Contrast-medium-induced nephropathy: is there a new consensus? A review of published guidelines

International Nuclear Information System (INIS)

Thomsen, Henrik S.; Morcos, Sameh K.

2006-01-01

The interest in contrast-medium-induced nephropathy has increased considerably during the last few years. Various guidelines regarding identifying patients at risk and measures to reduce the incidence of this complication have been proposed. The aim of this review was to analyse whether there is some consistency amongst these guidelines. A Medline search for the keyword ''contrast medium induced nephropathy'' during the period from the beginning of 2003 through the end of September 2005 was carried out. Only papers in English were reviewed. Thirteen guidelines were identified. Inconsistency was observed regarding advise on the prophylactic use of drugs and the isoosmolar dimer to reduce the incidence of contrast-medium-induced nephropathy. Consistency was found in relation to the importance of hydration, cessation of intake of nephrotoxic drugs and administration of the lowest possible dose of contrast medium. No new consensus has been observed in comparison to the European Society for Urogenital Radiology (ESUR) guidelines, which were published in 1999. (orig.)

Groomed jets in heavy-ion collisions: sensitivity to medium-induced bremsstrahlung

Energy Technology Data Exchange (ETDEWEB)

Mehtar-Tani, Yacine [Institute of Nuclear Theory, University of Washington,Seattle, WA 98195-1550 (United States); Tywoniuk, Konrad [Theoretical Physics Department, CERN,1211 Geneva 23 (Switzerland)

2017-04-21

We argue that contemporary jet substructure techniques might facilitate a more direct measurement of hard medium-induced gluon bremsstrahlung in heavy-ion collisions, and focus specifically on the “soft drop declustering” procedure that singles out the two leading jet substructures. Assuming coherent jet energy loss, we find an enhancement of the distribution of the energy fractions shared by the two substructures at small subjet energy caused by hard medium-induced gluon radiation. Departures from this approximation are discussed, in particular, the effects of colour decoherence and the contamination of the grooming procedure by soft background. Finally, we propose a complementary observable, that is the ratio of the two-pronged probability in Pb-Pb to proton-proton collisions and discuss its sensitivity to various energy loss mechanisms.

Groomed jets in heavy-ion collisions: sensitivity to medium-induced bremsstrahlung

CERN Document Server

Mehtar-Tani, Yacine

2017-04-21

We argue that contemporary jet substructure techniques might facilitate a more direct measurement of hard medium-induced gluon bremsstrahlung in heavy-ion collisions, and focus specifically on the "soft drop declustering" procedure that singles out the two leading jet substructures. Assuming coherent jet energy loss, we find an enhancement of the distribution of the energy fractions shared by the two substructures at small subjet energy caused by hard medium-induced gluon radiation. Departures from this approximation are discussed, in particular, the effects of colour decoherence and the contamination of the grooming procedure by soft background. Finally, we propose a complementary observable, that is the ratio of the two-pronged probability in Pb-Pb to proton-proton collisions and discuss its sensitivity to various energy loss mechanisms.

Groomed jets in heavy-ion collisions: sensitivity to medium-induced bremsstrahlung

Science.gov (United States)

Mehtar-Tani, Yacine; Tywoniuk, Konrad

2017-04-01

We argue that contemporary jet substructure techniques might facilitate a more direct measurement of hard medium-induced gluon bremsstrahlung in heavy-ion collisions, and focus specifically on the "soft drop declustering" procedure that singles out the two leading jet substructures. Assuming coherent jet energy loss, we find an enhancement of the distribution of the energy fractions shared by the two substructures at small subjet energy caused by hard medium-induced gluon radiation. Departures from this approximation are discussed, in particular, the effects of colour decoherence and the contamination of the grooming procedure by soft background. Finally, we propose a complementary observable, that is the ratio of the two-pronged probability in Pb-Pb to proton-proton collisions and discuss its sensitivity to various energy loss mechanisms.

Groomed jets in heavy-ion collisions: sensitivity to medium-induced bremsstrahlung

International Nuclear Information System (INIS)

Mehtar-Tani, Yacine; Tywoniuk, Konrad

2017-01-01

We argue that contemporary jet substructure techniques might facilitate a more direct measurement of hard medium-induced gluon bremsstrahlung in heavy-ion collisions, and focus specifically on the “soft drop declustering” procedure that singles out the two leading jet substructures. Assuming coherent jet energy loss, we find an enhancement of the distribution of the energy fractions shared by the two substructures at small subjet energy caused by hard medium-induced gluon radiation. Departures from this approximation are discussed, in particular, the effects of colour decoherence and the contamination of the grooming procedure by soft background. Finally, we propose a complementary observable, that is the ratio of the two-pronged probability in Pb-Pb to proton-proton collisions and discuss its sensitivity to various energy loss mechanisms.

Cellulose hydrolysis by fungi. 2. Cellulase production by Trichoderma harzianum in liquid medium fermentation

Energy Technology Data Exchange (ETDEWEB)

Roussos, S.; Raimbault, M. (Laboratoire de Microbiologie ORSTOM, Centre de Recherche IRCHA, 91 - Vert-le-Petit (France))

Microcrystalline cellulose (cellulose Avicel, Merck) supported growth of Trichoderma harzianum and induced production of cellulases in liquid cultures. After 50 h growth, the total cellulasic activities present in both the supernatant and the mycelium were 3,000 IU/l of carboxymethyl cellulose, 400 IU/l of filter paper activity, and 4 IU/l of cotton activity corresponding to 1.7 g/l of proteins. Cellulase production could be increased by a preliminary treatment of cellulose, and pH regulation during growth. The influence of inoculum concentration was studied and an optimum of 3 X 10/sup 7/ conidia/g dry weight of substrate was demonstrated. Using a synthetic culture medium, a soluble factor of germination was demonstrated which could be leached out by 3 successive washings of conidia.

Test procedures and instructions for Hanford complexant concentrate supernatant cesium removal using CST

Energy Technology Data Exchange (ETDEWEB)

Hendrickson, D.W.

1997-01-08

This document provides specific test procedures and instructions to implement the test plan for the preparation and conduct of a cesium removal test, using Hanford Complexant Concentrate supernatant liquor from tank 241-AN-107, in a bench-scale column. The cesium sorbent to be tested is crystalline silicotitanate. The test plan for which this provides instructions is WHC-SD-RE-TP-023, Hanford Complexant Concentrate Supernatant Cesium Removal Test Plan.

Event-by-event fluctuations in the medium-induced jet evolution

International Nuclear Information System (INIS)

Escobedo, Miguel A.; Iancu, Edmond

2016-01-01

We develop the event-by-event picture of the gluon distribution produced via medium-induced gluon branching by an energetic jet which propagates through a dense QCD medium. A typical event is characterized by the production of a large number of soft gluons which propagate at large angles with respect to the jet axis and which collectively carry a substantial amount of energy. By explicitly computing 2-gluon correlations, we demonstrate the existence of large event-by-event fluctuations, which reflect the stochastic nature of the branching process. For the two quantities that we have investigated — the energy loss at large angles and the soft gluon multiplicity —, the dispersion is parametrically as large as the respective expectation value. We identify interesting scaling laws, which suggest that the multiplicity distribution should exhibit KNO (Koba-Nielsen-Olesen) scaling. A similar scaling is known to hold for a jet branching in the vacuum, but the medium-induced distribution is found to be considerably broader. We predict that event-by-event measurements of the di-jet asymmetry in Pb+Pb collisions at the LHC should observe large fluctuations in the number of soft hadrons propagating at large angles and also in the total energy carried by these hadrons.

Cell-free culture supernatant of Bifidobacterium breve CNCM I-4035 decreases pro-inflammatory cytokines in human dendritic cells challenged with Salmonella typhi through TLR activation.

Science.gov (United States)

Bermudez-Brito, Miriam; Muñoz-Quezada, Sergio; Gomez-Llorente, Carolina; Matencio, Esther; Bernal, Maria J; Romero, Fernando; Gil, Angel

2013-01-01

Dendritic cells (DCs) constitute the first point of contact between gut commensals and our immune system. Despite growing evidence of the immunomodulatory effects of probiotics, the interactions between the cells of the intestinal immune system and bacteria remain largely unknown. Indeed,, the aim of this work was to determine whether the probiotic Bifidobacterium breve CNCM I-4035 and its cell-free culture supernatant (CFS) have immunomodulatory effects in human intestinal-like dendritic cells (DCs) and how they respond to the pathogenic bacterium Salmonella enterica serovar Typhi, and also to elucidate the molecular mechanisms involved in these interactions. Human DCs were directly challenged with B. breve/CFS, S. typhi or a combination of these stimuli for 4 h. The expression pattern of genes involved in Toll-like receptor (TLR) signaling pathway and cytokine secretion was analyzed. CFS decreased pro-inflammatory cytokines and chemokines in human intestinal DCs challenged with S. typhi. In contrast, the B. breve CNCM I-4035 probiotic strain was a potent inducer of the pro-inflammatory cytokines and chemokines tested, i.e., TNF-α, IL-8 and RANTES, as well as anti-inflammatory cytokines including IL-10. CFS restored TGF-β levels in the presence of Salmonella. Live B.breve and its supernatant enhanced innate immune responses by the activation of TLR signaling pathway. These treatments upregulated TLR9 gene transcription. In addition, CFS was a more potent inducer of TLR9 expression than the probiotic bacteria in the presence of S. typhi. Expression levels of CASP8 and IRAK4 were also increased by CFS, and both treatments induced TOLLIP gene expression. Our results indicate that the probiotic strain B. breve CNCM I-4035 affects the intestinal immune response, whereas its supernatant exerts anti-inflammatory effects mediated by DCs. This supernatant may protect immune system from highly infectious agents such as Salmonella typhi and can down-regulate pro

Cell-free culture supernatant of Bifidobacterium breve CNCM I-4035 decreases pro-inflammatory cytokines in human dendritic cells challenged with Salmonella typhi through TLR activation.

Directory of Open Access Journals (Sweden)

Miriam Bermudez-Brito

Full Text Available Dendritic cells (DCs constitute the first point of contact between gut commensals and our immune system. Despite growing evidence of the immunomodulatory effects of probiotics, the interactions between the cells of the intestinal immune system and bacteria remain largely unknown. Indeed,, the aim of this work was to determine whether the probiotic Bifidobacterium breve CNCM I-4035 and its cell-free culture supernatant (CFS have immunomodulatory effects in human intestinal-like dendritic cells (DCs and how they respond to the pathogenic bacterium Salmonella enterica serovar Typhi, and also to elucidate the molecular mechanisms involved in these interactions. Human DCs were directly challenged with B. breve/CFS, S. typhi or a combination of these stimuli for 4 h. The expression pattern of genes involved in Toll-like receptor (TLR signaling pathway and cytokine secretion was analyzed. CFS decreased pro-inflammatory cytokines and chemokines in human intestinal DCs challenged with S. typhi. In contrast, the B. breve CNCM I-4035 probiotic strain was a potent inducer of the pro-inflammatory cytokines and chemokines tested, i.e., TNF-α, IL-8 and RANTES, as well as anti-inflammatory cytokines including IL-10. CFS restored TGF-β levels in the presence of Salmonella. Live B.breve and its supernatant enhanced innate immune responses by the activation of TLR signaling pathway. These treatments upregulated TLR9 gene transcription. In addition, CFS was a more potent inducer of TLR9 expression than the probiotic bacteria in the presence of S. typhi. Expression levels of CASP8 and IRAK4 were also increased by CFS, and both treatments induced TOLLIP gene expression. Our results indicate that the probiotic strain B. breve CNCM I-4035 affects the intestinal immune response, whereas its supernatant exerts anti-inflammatory effects mediated by DCs. This supernatant may protect immune system from highly infectious agents such as Salmonella typhi and can down

Comparison of excretory-secretory antigen and positive faecal supernatant antigen in the detection of Echinococcus granulosus infection in dogs by CIEP

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P. R. Prathiush

Full Text Available Coproantigen detection of Echinococcosis in dogs by counter immunoelectrophoresis was standardized. Adult Echinococcus granulosus worms were obtained from intestine of a necropsied positive dog. Excretory-secretory antigen was prepared by culturing adult worms in Medium 199 (pH 7.4. Faeces of positive dog were collected and fecal supernatant was prepared and used for coproantigen detection. CIEP was carried out using tris-borate buffer (pH 8.0 at a constant current of 8mA/slide for 60 minutes. CIEP detected infection with both the antigens. [Vet World 2009; 2(11.000: 421-422

The effects of three types of macrophages culture supernatant on CFU-GM in irradiated mice

International Nuclear Information System (INIS)

Quan Hongxun; Fu Li; Zhao Fengchen; Han Fen

2008-01-01

Objective: To study the effects of peritional macrophyge(PM), alveolar macrophage (AM), and Kupffer cell (KC) on colony forming unite granulacyte/macrophage (CFU -GM) in irradiated mice. Methods: Using techniques of hemopoietic progenitors in vitro, the authors studied the effects of three types of macrophages culture supernatant on CFU - GM. Results: It is shown that three types of macrophages culture supernatant may stimulate proliferation and differentiation of CFU-GM in irradiated mice, and KC is the best one in comparison to others. Conclusion: three types of macrophages culture supernatant may protect CFU-GM irradiated mice with KC being the best method. (authors)

EspC, an Autotransporter Protein Secreted by Enteropathogenic Escherichia coli, Causes Apoptosis and Necrosis through Caspase and Calpain Activation, Including Direct Procaspase-3 Cleavage

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Antonio Serapio-Palacios

2016-06-01

Full Text Available Enteropathogenic Escherichia coli (EPEC has the ability to antagonize host apoptosis during infection through promotion and inhibition of effectors injected by the type III secretion system (T3SS, but the total number of these effectors and the overall functional relationships between these effectors during infection are poorly understood. EspC produced by EPEC cleaves fodrin, paxillin, and focal adhesion kinase (FAK, which are also cleaved by caspases and calpains during apoptosis. Here we show the role of EspC in cell death induced by EPEC. EspC is involved in EPEC-mediated cell death and induces both apoptosis and necrosis in epithelial cells. EspC induces apoptosis through the mitochondrial apoptotic pathway by provoking (i a decrease in the expression levels of antiapoptotic protein Bcl-2, (ii translocation of the proapoptotic protein Bax from cytosol to mitochondria, (iii cytochrome c release from mitochondria to the cytoplasm, (iv loss of mitochondrial membrane potential, (v caspase-9 activation, (vi cleavage of procaspase-3 and (vii an increase in caspase-3 activity, (viii PARP proteolysis, and (ix nuclear fragmentation and an increase in the sub-G1 population. Interestingly, EspC-induced apoptosis was triggered through a dual mechanism involving both independent and dependent functions of its EspC serine protease motif, the direct cleavage of procaspase-3 being dependent on this motif. This is the first report showing a shortcut for induction of apoptosis by the catalytic activity of an EPEC protein. Furthermore, this atypical intrinsic apoptosis appeared to induce necrosis through the activation of calpain and through the increase of intracellular calcium induced by EspC. Our data indicate that EspC plays a relevant role in cell death induced by EPEC.

Co-ordinate regulation of distinct host cell signalling pathways by multifunctional enteropathogenic Escherichia coli effector molecules.

Science.gov (United States)

Kenny, Brendan; Ellis, Sarah; Leard, Alan D; Warawa, Jonathan; Mellor, Harry; Jepson, Mark A

2002-05-01

Enteropathogenic Escherichia coli (EPEC) is a major cause of paediatric diarrhoea and a model for the family of attaching and effacing (A/E) pathogens. A/E pathogens encode a type III secretion system to transfer effector proteins into host cells. The EPEC Tir effector protein acts as a receptor for the bacterial surface protein intimin and is involved in the formation of Cdc42-independent, actin-rich pedestal structures beneath the adhered bacteria. In this paper, we demonstrate that EPEC binding to HeLa cells also induces Tir-independent, cytoskeletal rearrangement evidenced by the early, transient formation of filopodia-like structures at sites of infection. Filopodia formation is dependent on expression of the EPEC Map effector molecule - a protein that targets mitochondria and induces their dysfunction. We show that Map-induced filopodia formation is independent of mitochondrial targeting and is abolished by cellular expression of the Cdc42 inhibitory WASP-CRIB domain, demonstrating that Map has at least two distinct functions in host cells. The transient nature of the filopodia is related to an ability of EPEC to downregulate Map-induced cell signalling that, like pedestal formation, was dependent on both Tir and intimin proteins. The ability of Tir to downregulate filopodia was impaired by disrupting a putative GTPase-activating protein (GAP) motif, suggesting that Tir may possess such a function, with its interaction with intimin triggering this activity. Furthermore, we also found that Map-induced cell signalling inhibits pedestal formation, revealing that the cellular effects of Tir and Map must be co-ordinately regulated during infection. Possible implications of the multifunctional nature of EPEC effector molecules in pathogenesis are discussed.

Influence of Helicobacter pylori culture supernatant on the ecological balance of a dual-species oral biofilm.

Science.gov (United States)

Zhang, Wenling; Deng, Xiaohong; Zhou, Xuedong; Hao, Yuqing; Li, Yuqing

2018-01-01

Dental caries is a chronic progressive disease occurring in the tooth hard tissue due to multiple factors, in which bacteria are the initial cause. Both Streptococcus mutans and Streptococcus sanguinis are main members of oral biofilm. Helicobacter pylori may also be detected in dental plaque, playing an important role in the development of dental caries. The aim of this study was to investigate the effect of H. pylori culture supernatant on S. mutans and S. sanguinis dual-species biofilm and to evaluate its potential ability on affecting dental health. The effect of H. pylori supernatant on single-species and dual-species biofilm was measured by colony forming units counting and fluorescence in situ hybridization (FISH) assay, respectively. The effect of H. pylori supernatant on S. mutans and S. sanguinis extracellular polysaccharides (EPS) production was measured by both confocal laser scanning microscopy observation and anthrone-sulfuric acid method. The effect of H. pylori supernatant on S. mutans gene expression was measured by quantitative real-time PCR (qRT-PCR) assays. H. pylori supernatant could inhibit both S. mutans and S. sanguinis biofilm formation and EPS production. S. sanguinis inhibition rate was significantly higher than that of S. mutans. Finally, S. mutans bacteriocin and acidogenicity related genes expression were affected by H. pylori culture supernatant. Our results showed that H. pylori could destroy the balance between S. mutans and S. sanguinis in oral biofilm, creating an advantageous environment for S. mutans, which became the dominant bacteria, promoting the formation and development of dental caries.

Influence of Helicobacter pylori culture supernatant on the ecological balance of a dual-species oral biofilm

Directory of Open Access Journals (Sweden)

Wenling Zhang

2018-02-01

Full Text Available Abstract Dental caries is a chronic progressive disease occurring in the tooth hard tissue due to multiple factors, in which bacteria are the initial cause. Both Streptococcus mutans and Streptococcus sanguinis are main members of oral biofilm. Helicobacter pylori may also be detected in dental plaque, playing an important role in the development of dental caries. Objective The aim of this study was to investigate the effect of H. pylori culture supernatant on S. mutans and S. sanguinis dual-species biofilm and to evaluate its potential ability on affecting dental health. Material and methods The effect of H. pylori supernatant on single-species and dual-species biofilm was measured by colony forming units counting and fluorescence in situ hybridization (FISH assay, respectively. The effect of H. pylori supernatant on S. mutans and S. sanguinis extracellular polysaccharides (EPS production was measured by both confocal laser scanning microscopy observation and anthrone-sulfuric acid method. The effect of H. pylori supernatant on S. mutans gene expression was measured by quantitative real-time PCR (qRT-PCR assays. Results H. pylori supernatant could inhibit both S. mutans and S. sanguinis biofilm formation and EPS production. S. sanguinis inhibition rate was significantly higher than that of S. mutans. Finally, S. mutans bacteriocin and acidogenicity related genes expression were affected by H. pylori culture supernatant. Conclusion Our results showed that H. pylori could destroy the balance between S. mutans and S. sanguinis in oral biofilm, creating an advantageous environment for S. mutans, which became the dominant bacteria, promoting the formation and development of dental caries.

Nonlinear optical properties of an electromagnetically induced transparency medium interacting with two quantized fields

CERN Document Server

Kuang-Leman; Wu Yong Shi

2003-01-01

We study linear and nonlinear optical properties of an electromagnetically induced transparency (EIT) medium interacting with two quantized laser fields in the adiabatic EIT case. We show that the EIT medium exhibits normal dispersion. Kerr and higher-order nonlinear refractive index coefficients are also calculated in a completely analytical form. It is indicated that the EIT medium exhibits giant resonantly enhanced nonlinearities. We discuss the response of the EIT medium to nonclassical light fields and find that the polarization vanishes when the probe laser is initially in a nonclassical state of no single-photon coherence.

Patogénesis molecular, epidemiología y diagnóstico de Escherichia coli enteropatógena Molecular pathogenesis, epidemiology and diagnosis of enteropathogenic Escherichia coli

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Jorge E. Vidal

2007-10-01

Full Text Available Escherichia coli enteropatógena (EPEC es una de las principales causas de diarrea en niños menores de dos años en países en vías de desarrollo. La principal característica histopatológica de la infección es una lesión que induce la EPEC en el intestino conocida como la lesión A/E (adherencia y eliminación. Las bacterias se adhieren a los enterocitos y permiten la acumulación de la actina del citoesqueleto en la región apical de la célula, hasta formar una estructura de tipo "pedestal" y causar la eliminación de las microvellosidades intestinales. A pesar de que se conoce de modo detallado el proceso de formación de los pedestales de actina, aún no se ha esclarecido el mecanismo global de la diarrea que induce EPEC. La diarrea se ha vinculado con: a la destrucción de las microvellosidades del enterocito, b la salida masiva de iones hacia la luz intestinal y c la secreción de alguna enterotoxina. En estudios realizados en países en vías de desarrollo se ha demostrado que EPEC es uno de los principales agentes participantes en la diarrea infantil, con elevadas tasas de morbilidad y mortalidad. El diagnóstico microbiológico de la infección se realiza con metodologías adicionales a las utilizadas con regularidad en el laboratorio de microbiología clínica, entre ellas las siguientes: a serotipificación, b ensayo de adherencia, c prueba de FAS (tinción fluorescente para actina y d detección específica de genes que codifican a proteínas incluidas en la patogénesis, como el bfpA y eae. Un objetivo de esta revisión es actualizar los avances observados en la patogénesis molecular de la infección por EPEC, las metodologías para el diagnóstico microbiológico y la epidemiología en México y otros países en vías de desarrollo.Enteropathogenic Escherichia coli (EPEC is a leading cause of diarrhea in infants less than two years of age in developing countries. To induce diarrhea EPEC uses several virulence factors acting

The effect of fermented milk with Bifidobacterium infantis on ...

African Journals Online (AJOL)

STORAGESEVER

2008-11-19

Nov 19, 2008 ... In recent years, Bifidobacteria have attracted considerable attention due to their overall ... EPEC, the colonies were violet in colour with a metallic lustre on. EMB medium). ...... A new selective medium for. Bifidobacterium spp.

Aflatoxin B1 Degradation by Stenotrophomonas Maltophilia and Other Microbes Selected Using Coumarin Medium

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Tiangui Niu

2008-08-01

Full Text Available Aflatoxin B1 (AFB1 is one of the most harmful mycotoxins in animal production and food industry. A safe, effective and environmentally sound detoxification method is needed for controlling this toxin. In this study, 65 samples were screened from various sources with vast microbial populations using a newly developed medium containing coumarin as the sole carbon source. Twenty five single-colony bacterial isolates showing AFB1 reduction activity in a liquid culture medium were selected from the screen. Isolate 35-3, obtained from tapir feces and identified to be Stenotrophomonas maltophilia, reduced AFB1 by 82.5% after incubation in the liquid medium at 37 °C for 72 h. The culture supernatant of isolate 35-3 was able to degrade AFB1 effectively, whereas the viable cells and cell extracts were far less effective. Factors influencing AFB1 degradation by the culture supernatant were investigated. Activity was reduced to 60.8% and 63.5% at 20 °C and 30 °C, respectively, from 78.7% at 37 °C. The highest degradation rate was 84.8% at pH 8 and the lowest was only 14.3% at pH 4.0. Ions Mg2+ and Cu2+ were activators for AFB1 degradation, howeverï��Œion Zn2+ was a strong inhibitor. Treatments with proteinase K, proteinase K plus SDS and heating significantly reduced or eradicated the degradation activity of the culture supernatant. The results indicated that the degradation of AFB1 by S. maltophilia 35-3 was enzymatic and could have a great potential in industrial applications.

Use of QuantiFERON®-TB Gold in-tube culture supernatants for measurement of antibody responses.

Directory of Open Access Journals (Sweden)

Simon G Kimuda

Full Text Available QuantiFERON®-TB Gold in-tube (QFT-GIT supernatants may be important samples for use in assessment of anti-tuberculosis (TB antibodies when only limited volumes of blood can be collected and when a combination of antibody and cytokine measurements are required. These analytes, when used together, may also have the potential to differentiate active pulmonary TB (APTB from latent TB infection (LTBI. However, few studies have explored the use of QFT-GIT supernatants for investigations of antibody responses. This study determined the correlation and agreement between anti-CFP-10 and anti-ESAT-6 antibody concentrations in QFT-GIT nil supernatant and serum pairs from 68 TB household contacts. We also explored the ability of Mycobacterium tuberculosis (M.tb specific antibodies, or ratios of antibody to interferon gamma (IFN-γ in QFT-GIT supernatants, to differentiate 97 APTB cases from 58 individuals with LTBI. Sputum smear microscopy was used to define APTB, whereas the QFT-GIT and tuberculin skin test were used to define LTBI. There were strong and statistically significant correlations between anti-CFP-10 and anti-ESAT-6 antibodies in unstimulated QFT-GIT supernatants and sera (r = 0.89; p<0.0001 for both, and no significant differences in antibody concentration between them. Anti-CFP-10 & anti-ESAT-6 antibodies differentiated APTB from LTBI with sensitivities of 88.7% & 71.1% and specificities of 41.4% & 51.7% respectively. Anti-CFP-10 antibody/M.tb specific IFN-γ and anti-ESAT-6 antibody/M.tb specific IFN-γ ratios had sensitivities of 48.5% & 54.6% and specificities of 89.7% and 75.9% respectively. We conclude that QFT-GIT nil supernatants may be used in the place of sera when measuring antibody responses, reducing blood volumes needed for such investigations. Antibodies in QFT-GIT nil supernatants on their own discriminate APTB from LTBI with high sensitivity but have poor specificity, whereas the reverse is true when antibodies are used in

The Effect of Anaerobic and Aerobic Fish Sludge Supernatant on Hydroponic Lettuce

Directory of Open Access Journals (Sweden)

Simon Goddek

2016-06-01

Full Text Available The mobilization of nutrients from fish sludge (i.e., feces and uneaten feed plays a key role in optimizing the resource utilization and thus in improving the sustainability of aquaponic systems. While several studies have documented the aerobic and anaerobic digestion performance of aquaculture sludge, the impact of the digestate on plant growth has yet to be understood. The present study examines the impact of either an aerobic or an anaerobic digestion effluent on lettuce plant growth, by enriching a mixture of aquaculture and tap water with supernatants from both aerobic and anaerobic batch reactors. The lettuce plants grown in the hydroponic system supplied with supernatant from an anaerobic reactor had significantly better performance with respect to weight gain than both, those in the system where supernatant from the aerobic reactor was added, as well as the control system. It can be hypothesized that this effect was caused by the presence of NH4+ as well as dissolved organic matter, plant growth promoting rhizobacteria and fungi, and humic acid, which are predominantly present in anaerobic effluents. This study should therefore be of value to researchers and practitioners wishing to further develop sludge remineralization in aquaponic systems.

Ultrafiltration and Characterization of AW-101 Supernatant and Entrained Solids

International Nuclear Information System (INIS)

Brooks, K.P.; Bredt, P.R.; Golcar, G.R.; Hartley, S.A.; Urie, M.W.; Tingey, J.M.; Rappe, K.G.; Jagoda, L.K.

1999-01-01

The River Protection Project Waste Treatment Plant (RPP-WTP) (1996) flow sheet uses cross-flow filtration as the solid/liquid separation technique. Unlike traditional dead-end filtration, which has a declining rate caused by the growth of a filter cake on the surface of the filter medium, in cross-flow filtration, the filter cake is swept away by the fluid flowing across it. This filtration method is especially beneficial when there are very fine particles and when system simplicity is required. The objective of this work was to test cross-flow filtration using actual Envelope A Hanford tank waste. Similar to the Phase 1A study, they evaluated the permeability of an Envelope A feed through a single element filter as a function of transmembrane pressure, axial velocity, solids concentration, and time. In addition, the efficiency of back pulse and chemical cleaning on the filter performance was evaluated. The chemical and radiochemical composition of the filtrate and solids was measured to determine efficiency of the filtration process. This report describes the test apparatus, the experimental approach, the results of the tests, and the chemical and radiochemical analysis for supernatants taken from Hanford Tank AW-101. This report also provides a means of transmitting to British Nuclear Fuels, Limited (BNFL) the completed test instruction and raw filtration and analytical data

Conditioned medium from alternatively activated macrophages induce mesangial cell apoptosis via the effect of Fas

International Nuclear Information System (INIS)

Huang, Yuan; Luo, Fangjun; Li, Hui; Jiang, Tao; Zhang, Nong

2013-01-01

During inflammation in the glomerulus, the proliferation of myofiroblast-like mesangial cells is commonly associated with the pathological process. Macrophages play an important role in regulating the growth of resident mesangial cells in the glomeruli. Alternatively activated macrophage (M2 macrophage) is a subset of macrophages induced by IL-13/IL-4, which is shown to play a repair role in glomerulonephritis. Prompted by studies of development, we performed bone marrow derived macrophage and rat mesangial cell co-culture study. Conditioned medium from IL-4 primed M2 macrophages induced rat mesangial cell apoptosis. The pro-apoptotic effect of M2 macrophages was demonstrated by condensed nuclei stained with Hoechst 33258, increased apoptosis rates by flow cytometry analysis and enhanced caspase-3 activation by western blot. Fas protein was up-regulated in rat mesangial cells, and its neutralizing antibody ZB4 partly inhibited M2 macrophage-induced apoptosis. The up-regulated arginase-1 expression in M2 macrophage also contributed to this apoptotic effect. These results indicated that the process of apoptosis triggered by conditioned medium from M2 macrophages, at least is partly conducted through Fas in rat mesangial cells. Our findings provide compelling evidence that M2 macrophages control the growth of mesangial cells in renal inflammatory conditions. - Highlights: • Conditioned-medium from M2 macrophages induces rat mesangial cell (MsC) apoptosis. • M2 macrophage conditioned medium exerts its pro-apoptotic effects via Fas ligand. • Arginase-1 activity in M2 macrophages plays a role in inducing apoptosis in rat MsC

Conditioned medium from alternatively activated macrophages induce mesangial cell apoptosis via the effect of Fas

Energy Technology Data Exchange (ETDEWEB)

Huang, Yuan; Luo, Fangjun; Li, Hui; Jiang, Tao; Zhang, Nong, E-mail: nzhang@fudan.edu.cn

2013-11-15

During inflammation in the glomerulus, the proliferation of myofiroblast-like mesangial cells is commonly associated with the pathological process. Macrophages play an important role in regulating the growth of resident mesangial cells in the glomeruli. Alternatively activated macrophage (M2 macrophage) is a subset of macrophages induced by IL-13/IL-4, which is shown to play a repair role in glomerulonephritis. Prompted by studies of development, we performed bone marrow derived macrophage and rat mesangial cell co-culture study. Conditioned medium from IL-4 primed M2 macrophages induced rat mesangial cell apoptosis. The pro-apoptotic effect of M2 macrophages was demonstrated by condensed nuclei stained with Hoechst 33258, increased apoptosis rates by flow cytometry analysis and enhanced caspase-3 activation by western blot. Fas protein was up-regulated in rat mesangial cells, and its neutralizing antibody ZB4 partly inhibited M2 macrophage-induced apoptosis. The up-regulated arginase-1 expression in M2 macrophage also contributed to this apoptotic effect. These results indicated that the process of apoptosis triggered by conditioned medium from M2 macrophages, at least is partly conducted through Fas in rat mesangial cells. Our findings provide compelling evidence that M2 macrophages control the growth of mesangial cells in renal inflammatory conditions. - Highlights: • Conditioned-medium from M2 macrophages induces rat mesangial cell (MsC) apoptosis. • M2 macrophage conditioned medium exerts its pro-apoptotic effects via Fas ligand. • Arginase-1 activity in M2 macrophages plays a role in inducing apoptosis in rat MsC.

Medium-Throughput Screen of Microbially Produced Serotonin via a G-Protein-Coupled Receptor-Based Sensor.

Science.gov (United States)

Ehrenworth, Amy M; Claiborne, Tauris; Peralta-Yahya, Pamela

2017-10-17

Chemical biosensors, for which chemical detection triggers a fluorescent signal, have the potential to accelerate the screening of noncolorimetric chemicals produced by microbes, enabling the high-throughput engineering of enzymes and metabolic pathways. Here, we engineer a G-protein-coupled receptor (GPCR)-based sensor to detect serotonin produced by a producer microbe in the producer microbe's supernatant. Detecting a chemical in the producer microbe's supernatant is nontrivial because of the number of other metabolites and proteins present that could interfere with sensor performance. We validate the two-cell screening system for medium-throughput applications, opening the door to the rapid engineering of microbes for the increased production of serotonin. We focus on serotonin detection as serotonin levels limit the microbial production of hydroxystrictosidine, a modified alkaloid that could accelerate the semisynthesis of camptothecin-derived anticancer pharmaceuticals. This work shows the ease of generating GPCR-based chemical sensors and their ability to detect specific chemicals in complex aqueous solutions, such as microbial spent medium. In addition, this work sets the stage for the rapid engineering of serotonin-producing microbes.

Bacterial Population in Intestines of Litopenaeus vannamei Fed Different Probiotics or Probiotic Supernatant.

Science.gov (United States)

Sha, Yujie; Liu, Mei; Wang, Baojie; Jiang, Keyong; Qi, Cancan; Wang, Lei

2016-10-28

The interactions of microbiota in the gut play an important role in promoting or maintaining the health of hosts. In this study, in order to investigate and compare the effects of dietary supplementation with Lactobacillus pentosus HC-2 (HC-2), Enterococcus faecium NRW-2, or the bacteria-free supernatant of a HC-2 culture on the bacterial composition of Litopenaeus vannamei , Illumina sequencing of the V1-V2 region of the 16S rRNA gene was used. The results showed that unique species exclusively existed in specific dietary groups, and the abundance of Actinobacteria was significantly increased in the intestinal bacterial community of shrimp fed with the bacteria-free supernatant of an HC-2 culture compared with the control. In addition, the histology of intestines of the shrimp from the four dietary groups was also described, but no obvious improvements in the intestinal histology were observed. The findings in this work will help to promote the understanding of the roles of intestinal bacteria in shrimps when fed with probiotics or probiotic supernatant.

Experimental data developed to support the selection of a treatment process for West Valley alkaline supernatant

Energy Technology Data Exchange (ETDEWEB)

Bray, L.A.; Holton, L.K.; Myers, T.R.; Richardson, G.M.; Wise, B.M.

1984-01-01

At the request of West Valley Nuclear Services Co., Inc., the Pacific Northwest Laboratory (PNL) has studied alternative treatment processes for the alkaline PUREX waste presently being stored in Tank 8D2 at West Valley, New York. Five tasks were completed during FY 1983: (1) simulation and characterization of the alkaline supernatant and sludge from the tank. The radiochemical and chemical distributions between the aqueous and solid phase were determined, and the efficiency of washing sludge with water to remove ions such as Na/sup +/ and SO/sub 4//sup 2 -/ was investigated; (2) evaluation of a sodium tetraphenylboron (Na-TPB) precipitation process to recover cesium (Cs) and a sodium titanate (Na-TiA) sorption process to recover strontium (Sr) and plutonium (Pu) from the West Valley Alkaline supernatant. These processes were previously developed and tested at the US Department of Energy's Savannah River Plant; (3) evaluation of an organic cation-exchange resin (Duolite CS-100) to recover Cs and Pu from the alkaline supernatant followed by an organic macroreticular cation exchange resin (Amberlite IRC-718) to recover Sr; (4) evaluation of an inorganic ion exchanger (Linde Ionsiv IE-95) to recover Cs, Sr, and Pu from the alkaline supernatant; and (5) evaluation of Dowex-1,X8 organic anion exchange resin to recover technetium (Tc) from alkaline supernatant. The findings of these tasks are reported. 21 references, 36 figures, 34 tables.

Culture Supernatants of Lactobacillus gasseri and L. crispatus Inhibit Candida albicans Biofilm Formation and Adhesion to HeLa Cells.

Science.gov (United States)

Matsuda, Yuko; Cho, Otomi; Sugita, Takashi; Ogishima, Daiki; Takeda, Satoru

2018-03-30

Vulvovaginal candidiasis (VVC) is a common superficial infection of the vaginal mucous membranes caused by the fungus Candida albicans. The aim of this study was to assess the mechanisms underlying the inhibitory effects of the culture supernatants of Lactobacillus gasseri and L. crispatus, the predominant microbiota in Asian healthy women, on C. albicans biofilm formation. The inhibition of C. albicans adhesion to HeLa cells by Lactobacillus culture supernatant was also investigated. Candida albicans biofilm was formed on polystyrene flat-bottomed 96-well plates, and the inhibitory effects on the initial colonization and maturation phases were determined using the XTT reduction assay. The expression levels of biofilm formation-associated genes (HWP1, ECE1, ALS3, BCR1, EFG1, TEC1, and CPH1) were determined by reverse transcription quantitative polymerase chain reaction. The inhibition of C. albicans adhesion to HeLa cells by Lactobacillus culture supernatant was evaluated by enumerating viable C. albicans cells. The culture supernatants of both Lactobacillus species inhibited the initial colonization and maturation of C. albicans biofilm. The expression levels of all biofilm formation-related genes were downregulated in the presence of Lactobacillus culture supernatant. The culture supernatant also inhibited C. albicans adhesion to HeLa cells. The culture supernatants of L. gasseri and L. crispatus inhibited C. albicans biofilm formation by downregulating biofilm formation-related genes and C. albicans adhesion to HeLa cells. These findings support the notion that Lactobacillus metabolites may be useful alternatives to antifungal drugs for the management of VVC.

Cytokine profiles of tumor supernatants in invasive ductal cancer and fibroadenoma of the breast and its relationship with VEGF-A expression in the tumors.

Science.gov (United States)

Autenshlyus, Alexander I; Arkhipov, Sergey A; Kunts, Tatiana A; Marinkin, Igor O; Mikhailova, Elena S; Karpukhina, Xenia V; Varaksin, Nikolay A

2017-03-01

Interrelations between cytokines, produced by invasive ductal carcinoma (IDC) and fibroadenoma (FA) of the breast, and angiogenic growth factor VEGF-A, expressed in IDC and FA, were investigated. The analysis of the cytokine profiles of IDC and FA was performed by cultivation of tumor biopsy specimens in vitro. Testing of the cytokine-producing reserve of the tumors for production of VEGF-A was conducted by culturing samples of IDC and FA in a medium containing polyclonal activator (a complex of phytohemagglutinin, concanavalin A, and lipopolysaccharide). Levels of cytokines and growth factors (IL-2, IL-6, IL-8, IL-10, IL-17, IL-18, IL-1β, IL-1Ra, TNF-α, IFN-γ, G-CSF, GM-CSF, VEGF-A) and MCP-1 (monocyte chemoattractant protein-1) in tumor supernatants were determined by an ELISA. Expression of VEGF-A was analyzed in tumor biopsy specimens by immunohistochemical analysis. In the IDC supernatants, the concentrations of IL-17, IL-18, and IFN-γ were higher and the concentrations of IL-10 and MCP-1 were lower in comparison with the FA supernatants. We observed negative correlations between the macrophage infiltration and VEGF-A concentration in the IDC supernatants (r = -0.508; P = 0.011) and between VEGF-A expression and the IDC vascularization degree (r = -0.423, P = 0.039). Spontaneous expression of VEGF-A in samples of IDC significantly exceeded the VEGF-A expression in FA. There was no difference between IDC and FA in VEGF-A expression after treatment with the polyclonal activators. Our results indicate that greater malignancy may have a paradoxical effect that is controlled by cytokines and characterized by weakening of tumor angiogenesis during overproduction of VEGF-A. These findings point to complex mechanisms of positive and negative regulation of tumor angiogenesis by cytokines that are produced by the tumor and by cells in its microenvironment, whose cytokine profiles may change at different stages of tumor progression.

Aggravation of cold-induced injury in Vero-B4 cells by RPMI 1640 medium – Identification of the responsible medium components

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Pless-Petig Gesine

2012-10-01

Full Text Available Abstract Background In modern biotechnology, there is a need for pausing cell lines by cold storage to adapt large-scale cell cultures to the variable demand for their products. We compared various cell culture media/solutions for cold storage of Vero-B4 kidney cells, a cell line widely used in biotechnology. Results Cold storage in RPMI 1640 medium, a recommended cell culture medium for Vero-B4 cells, surprisingly, strongly enhanced cold-induced cell injury in these cells in comparison to cold storage in Krebs-Henseleit buffer or other cell culture media (DMEM, L-15 and M199. Manufacturer, batch, medium supplements and the most likely components with concentrations outside the range of the other media/solutions (vitamin B12, inositol, biotin, p-aminobenzoic acid did not cause this aggravation of cold-induced injury in RPMI 1640. However, a modified Krebs-Henseleit buffer with a low calcium concentration (0.42 mM, a high concentration of inorganic phosphate (5.6 mM, and glucose (11.1 mM; i.e. concentrations as in RPMI 1640 evoked a cell injury and loss of metabolic function corresponding to that observed in RPMI 1640. Deferoxamine improved cell survival and preserved metabolic function in modified Krebs-Henseleit buffer as well as in RPMI 1640. Similar Ca2+ and phosphate concentrations did not increase cold-induced cell injury in the kidney cell line LLC-PK1, porcine aortic endothelial cells or rat hepatocytes. However, more extreme conditions (Ca2+ was nominally absent and phosphate concentration raised to 25 mM as in the organ preservation solution University of Wisconsin solution also increased cold-induced injury in rat hepatocytes and porcine aortic endothelial cells. Conclusion These data suggest that the combination of low calcium and high phosphate concentrations in the presence of glucose enhances cold-induced, iron-dependent injury drastically in Vero-B4 cells, and that a tendency for this pathomechanism also exists in other cell types.

Neutrophil-induced transmigration of tumour cells treated with tumour-conditioned medium is facilitated by granulocyte-macrophage colony-stimulating factor.

LENUS (Irish Health Repository)

Wu, Q D

2012-02-03

OBJECTIVE: To investigate the effect of different cytokines that are present in tumour-conditioned medium on human neutrophil (PMN)-induced tumour cell transmigration. DESIGN: Laboratory study. SETTING: University hospital, Ireland. MATERIAL: Isolated human PMN and cultured human breast tumour cell line, MDA-MB-231. Interventions: Human PMN treated with either tumour-conditioned medium or different media neutralised with monoclonal antibodies (MoAb), and MDA-MB-231 cells were plated on macrovascular and microvascular endothelial monolayers in collagen-coated transwells to assess migration of tumour cells. MAIN OUTCOME MEASURES: Cytokines present in tumour-conditioned medium, PMN cytocidal function and receptor expression, and tumour cell transmigration. RESULTS: tumour-conditioned medium contained high concentrations of granulocyte-macrophage colony-stimulating factor (GM-CSF), vascular endothelial growth factor (VEGF), and interleukin 8 (IL-8), but not granulocyte colony-stimulating factor (G-CSF) and interleukin 3 (IL-3). Anti-GM-CSF MoAb significantly reduced PMN-induced transmigration of tumour cells treated with tumour-conditioned medium (p < 0.05), whereas anti-VEGF and anti-IL-8 MoAbs did not affect their migration. In addition, anti-GM-CSF MoAb, but not anti-VEGF or anti-IL-8 MoAb, reduced PMN CD11b and CD18 overexpression induced by tumour-conditioned medium (p < 0.05). CONCLUSION: These results indicate that the GM-CSF that is present in tumour-conditioned medium may be involved, at least in part, in alterations in PMN function mediated by the medium and subsequently PMN-induced transmigration of tumour cells.

Medium-induced gluon radiation and colour decoherence beyond the soft approximation

CERN Document Server

Apolinário, Liliana; Milhano, José Guilherme; Salgado, Carlos A

2015-01-01

We derive the in-medium gluon radiation spectrum off a quark within the path integral formalism at finite energies, including all next-to-eikonal corrections in the propagators of quarks and gluons. Results are computed for finite formation times, including interference with vacuum amplitudes. Rewriting the medium averages in a convenient manner we present the spectrum in terms of dipole cross sections and a colour decoherence parameter with the same physical origin as that found in previous studies of the antenna radiation. This factorisation allows us to present a simple physical picture of the medium-induced radiation for any value of the formation time, of interest for a probabilistic implementation of the modified parton shower. Interestingly -- and unexpectedly -- we also find a modification of the contribution from the hard vertex which cannot be factorized, at finite formation time, as the vacuum Altarelli-Parisi splitting function. Known results are recovered for the particular cases of soft radiatio...

Products of cells from gliomas: VIII. Multiple-well immunoperoxidase assay of immunoreactivity of primary hybridoma supernatants with human glioma and brain tissue and cultured glioma cells.

Science.gov (United States)

McKeever, P E; Wahl, R L; Shakui, P; Jackson, G A; Letica, L H; Liebert, M; Taren, J A; Beierwaltes, W H; Hoff, J T

1990-06-01

To test the feasibility of primary screening of hybridoma supernatants against human glioma tissue, over 5000 combinations of hybridoma supernatants with glioma tissue, cultured glioma cells, and normal central neural tissue were screened with a new multiple-well (M-well) screening system. This is an immunoperoxidase assay system with visual endpoints for screening 20-30 hybridoma supernatants per single microscope slide. There were extensive differences between specificities to tissue and to cultured glioma cells when both were screened with M-wells and when cultured cells were screened with standard semi-automated fluorescence. Primary M-well screening with glioma tissue detected seven hybridoma supernatants that specifically identified parenchymal cells of glioma tissue and that were not detected with cultured cells. Immunoreactivities of individual supernatants for vascular components (nine supernatants), necrosis (five supernatants), and nuclei (three supernatants) were detected. Other supernatants bound multiple sites on glioma tissue and/or subpopulations of neurons and glia of normal tissue. The results show that primary screening with glioma tissue detects a number of different specificities of hybridoma supernatants to gliomas not detected by conventional screening with cultured cells. These are potentially applicable to diagnosis and therapy.

Medium from X-rayed cultures induces DNA strand-breaks in non-irradiated HeLa cells

International Nuclear Information System (INIS)

Ikushima, T.; Okuyama, K.; Tanizaki, Y.

2002-01-01

There is growing evidence to indicate that several types of responses are induced by ionizing radiation in non-irradiated cells. Such bystander effects include the killing of non-irradiated cells, the induction of sister chromatid exchanges and chromosomal aberrations, and the induction of gene mutations and chromosomal instability and enhanced cell growth. In the present study, we assessed whether the medium from irradiated cultures can induce DNA strand-breaks in non-irradiated cells, using single-cell gel electrophoresis assay (comet assay). HeLa cells in culture were irradiated with 0.5 to 8 Gy of 140 kVp X-rays and one hour later, the medium was taken from the irradiated culture, passed through a filter and transferred to the parallel culture of non-irradiated HeLa cells as non-target cells. After incubation for 30 min, the comet assay was performed under alkaline and neutral conditions. Such treatments resulted in a dose-dependent increase in tail moment under either alkaline or neutral condition, indicating the induction of DNA single- or double-strand breaks, respectively. It was also shown that the clonogenic survival was reduced in the cells cultured in the medium from irradiated cultures. Such a change was not detected at all when medium alone was irradiated. These results provided disputed evidence that irradiated cells released certain genotoxic factor(s) into the culture medium that can induce DNA strand breaks leading to cell death. Our results suggest that physical contact between irradiated and non-irradiated cells may not be necessary for the bystander effects observed in this study. It appears that bystander responses may be mediated by multiple mechanisms

Cement waste form qualification report: WVDP [West Valley Demonstration Project] PUREX decontaminated supernatant

International Nuclear Information System (INIS)

McVay, C.W.; Stimmel, J.R.; Marchetti, S.

1988-08-01

This report provides a summary of work performed to develop a cement-based, low-level waste formulation suitable for the solidification of decontaminated high-level waste liquid produced as a by-product of PUREX spent fuel reprocessing. The resultant waste form is suitable for interim storage and is intended for ultimate disposal as low-level Class C waste; it also meets the stability requirements of the NRC Branch Technical Position on Waste Form Qualification, May 1983 and the requirements of 10 CFR 61. A recipe was developed utilizing only Portland Type I cement based on an inorganic salts simulant of the PUREX supernatant. The qualified recipe was tested full scale in the production facility and was observed to produce a product with entrained air, low density, and lower-than-expected compressive strength. Further laboratory scale testing with actual decontaminated supernatant revealed that set retarders were present in the supernatant, precluding setting of the product and allowing the production of ''bleed water.'' Calcium nitrate and sodium silicate were added to overcome the set retarding effect and produced a final product with improved performance when compared to the original formulation. This report describes the qualification process and qualification test results for the final product formulation. 7 refs., 38 figs., 21 tabs

Protease signaling through protease activated receptor 1 mediate nerve activation by mucosal supernatants from irritable bowel syndrome but not from ulcerative colitis patients.

Science.gov (United States)

Buhner, Sabine; Hahne, Hannes; Hartwig, Kerstin; Li, Qin; Vignali, Sheila; Ostertag, Daniela; Meng, Chen; Hörmannsperger, Gabriele; Braak, Breg; Pehl, Christian; Frieling, Thomas; Barbara, Giovanni; De Giorgio, Roberto; Demir, Ihsan Ekin; Ceyhan, Güralp Onur; Zeller, Florian; Boeckxstaens, Guy; Haller, Dirk; Kuster, Bernhard; Schemann, Michael

2018-01-01

The causes of gastrointestinal complaints in irritable bowel syndrome (IBS) remain poorly understood. Altered nerve function has emerged as an important pathogenic factor as IBS mucosal biopsy supernatants consistently activate enteric and sensory neurons. We investigated the neurally active molecular components of such supernatants from patients with IBS and quiescent ulcerative colitis (UC). Effects of supernatants from 7 healthy controls (HC), 20 IBS and 12 UC patients on human and guinea pig submucous neurons were studied with neuroimaging techniques. We identify differentially expressed proteins with proteome analysis. Nerve activation by IBS supernatants was prevented by the protease activated receptor 1 (PAR1) antagonist SCHE79797. UC supernatants also activated enteric neurons through protease dependent mechanisms but without PAR1 involvement. Proteome analysis of the supernatants identified 204 proteins, among them 17 proteases as differentially expressed between IBS, UC and HC. Of those the four proteases elastase 3a, chymotrypsin C, proteasome subunit type beta-2 and an unspecified isoform of complement C3 were significantly more abundant in IBS compared to HC and UC supernatants. Of eight proteases, which were upregulated in IBS, the combination of elastase 3a, cathepsin L and proteasome alpha subunit-4 showed the highest prediction accuracy of 98% to discriminate between IBS and HC groups. Elastase synergistically potentiated the effects of histamine and serotonin-the two other main neuroactive substances in the IBS supernatants. A serine protease inhibitor isolated from the probiotic Bifidobacterium longum NCC2705 (SERPINBL), known to inhibit elastase-like proteases, prevented nerve activation by IBS supernatants. Proteases in IBS and UC supernatants were responsible for nerve activation. Our data demonstrate that proteases, particularly those signalling through neuronal PAR1, are biomarker candidates for IBS, and protease profiling may be used to

Optical illusions induced by rotating medium

Science.gov (United States)

Zang, XiaoFei; Huang, PengCheng; Zhu, YiMing

2018-03-01

Different from the traditional single-function electromagnetic wave rotators (rotate the electromagnetic wavefronts), we propose that rotating medium can be extended to optical illusions such as breaking the diffraction limit and overlapping illusion. Furthermore, the homogeneous but anisotropic rotating medium is simplified by homogeneous and isotropic positive-index materials according to the effective medium theory, which is helpful for future device fabrication. Finite element simulations for the two-dimensional case are performed to demonstrate these properties.

Large Fizeau's light-dragging effect in a moving electromagnetically induced transparent medium.

Science.gov (United States)

Kuan, Pei-Chen; Huang, Chang; Chan, Wei Sheng; Kosen, Sandoko; Lan, Shau-Yu

2016-10-03

As one of the most influential experiments on the development of modern macroscopic theory from Newtonian mechanics to Einstein's special theory of relativity, the phenomenon of light dragging in a moving medium has been discussed and observed extensively in different types of systems. To have a significant dragging effect, the long duration of light travelling in the medium is preferred. Here we demonstrate a light-dragging experiment in an electromagnetically induced transparent cold atomic ensemble and enhance the dragging effect by at least three orders of magnitude compared with the previous experiments. With a large enhancement of the dragging effect, we realize an atom-based velocimeter that has a sensitivity two orders of magnitude higher than the velocity width of the atomic medium used. Such a demonstration could pave the way for motional sensing using the collective state of atoms in a room temperature vapour cell or solid state material.

The influence of medium elasticity on the prediction of histotripsy-induced bubble expansion and erythrocyte viability

Science.gov (United States)

Bader, Kenneth B.

2018-05-01

Histotripsy is a form of therapeutic ultrasound that liquefies tissue mechanically via acoustic cavitation. Bubble expansion is paramount in the efficacy of histotripsy therapy, and the cavitation dynamics are strongly influenced by the medium elasticity. In this study, an analytic model to predict histotripsy-induced bubble expansion in a fluid was extended to include the effects of medium elasticity. Good agreement was observed between the predictions of the analytic model and numerical computations utilizing highly nonlinear excitations (shock-scattering histotripsy) and purely tensile pulses (microtripsy). No bubble expansion was computed for either form of histotripsy when the elastic modulus was greater than 20 MPa and the peak negative pressure was less than 50 MPa. Strain in the medium due to the expansion of a single bubble was also tabulated. The viability of red blood cells was calculated as a function of distance from the bubble wall based on empirical data of impulsive stretching of erythrocytes. Red blood cells remained viable at distances further than 44 µm from the bubble wall. As the medium elasticity increased, the distance over which bubble expansion-induced strain influenced red blood cells was found to decrease sigmoidally. These results highlight the relationship between tissue elasticity and the efficacy of histotripsy. In addition, an upper medium elasticity limit was identified, above which histotripsy may not be effective for tissue liquefaction.

Medium Modifications of Hadrons in Photon Induced Reactions

International Nuclear Information System (INIS)

Schadmand, S.

2004-01-01

Indications for in-medium modifications of hadron properties are reported from photoabsorption and meson production experiments. Strong medium modifications are observed in inclusive photoabsorption experiments and theoretical models investigate the in-medium dynamics of baryon resonances and their coupling to mesons. Recent experiments study the in-medium behavior of scalar and vector mesons where theoretical models expect in-medium modifications of the meson spectral functions that might be connected to partial restoration of chiral symmetry

Properties of Two Novel Esterases Identified from Culture Supernatant of Penicillium purpurogenum Grown on Sugar Beet Pulp.

Science.gov (United States)

Oleas, Gabriela; Callegari, Eduardo; Sepulveda, Romina; Eyzaguirre, Jaime

2016-01-01

The filamentous fungus Penicillium purpurogenum grows on a variety of natural carbon sources, such as sugar beet pulp, and secretes to the medium a large number of enzymes that degrade the carbohydrate components of lignocellulose. Sugar beet pulp is rich in pectin, and the purpose of this work is to identify novel esterases produced by the fungus, which may participate in pectin degradation. Partially purified culture supernatants of the fungus grown on sugar beet pulp were subjected to mass spectrometry analysis. Peptides thus identified, which may be part of potential esterases were probed against the proteins deduced from the fungal genome sequence. The cDNAs of two putative esterases identified were expressed in Pichia pastoris and their properties studied. One of these enzymes, named FAET, is a feruloyl esterase, while the other, PE, is classified as a pectin methyl esterase. These findings add to our knowledge of the enzymology of pectin degradation by Penicillium purpurogenum, and define properties of two novel esterases acting on de-esterification of pectin. Their availability may be useful as tools for the study of pectin structure and degradation.

Design of an efficient medium for heterologous protein production in Yarrowia lipolytica: case of human interferon alpha 2b.

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Gasmi, Najla; Ayed, Atef; Nicaud, Jean-Marc; Kallel, Héla

2011-05-20

The non conventional yeast Yarrowia lipolytica has aroused a strong industrial interest for heterologous protein production. However most of the studies describing recombinant protein production by this yeast rely on the use of complex media, such media are not convenient for large scale production particularly for products intended for pharmaceutical applications. In addition medium composition can also affect the production yield. Hence it is necessary to design an efficient medium for therapeutic protein expression by this host. Five different media, including four minimal media and a complex medium, were assessed in shake flasks for the production of human interferon alpha 2b (hIFN α2b) by Y. lipolytica under the control of POX2 promoter inducible with oleic acid. The chemically defined medium SM4 formulated by Invitrogen for Pichia pastoris growth was the most suitable. Using statistical experimental design this medium was further optimized. The selected minimal medium consisting in SM4 supplemented with 10 mg/l FeCl₃, 1 g/l glutamate, 5 ml/l PTM1 (Pichia Trace Metals) solution and a vitamin solution composed of myo-inositol, thiamin and biotin was called GNY medium. Compared to shake flask, bioreactor culture in GNY medium resulted in 416-fold increase of hIFN α2b production and 2-fold increase of the biological activity. Furthermore, SM4 enrichment with 5 ml/l PTM1 solution contributed to protect hIFN α2b against the degradation by the 28 kDa protease identified by zymography gel in culture supernatant. The screening of the inhibitory effect of the trace elements present in PTM1 solution on the activity of this protease was achieved using a Box-Behnken design. Statistical data analysis showed that FeCl₃ and MnSO₄ had the most inhibitory effect. We have designed an efficient medium for large scale production of heterologous proteins by Y. lipolytica. The optimized medium GNY is suitable for the production of hIFN α2b with the advantage that no

Underestimation of phosphorus fraction change in the supernatant after phosphorus adsorption onto iron oxides and iron oxide-natural organic matter complexes.

Science.gov (United States)

Yan, Jinlong; Jiang, Tao; Yao, Ying; Wang, Jun; Cai, Yuanli; Green, Nelson W; Wei, Shiqiang

2017-05-01

The phosphorus (P) fraction distribution and formation mechanism in the supernatant after P adsorption onto iron oxides and iron oxide-humic acid (HA) complexes were analyzed using the ultrafiltration method in this study. With an initial P concentration of 20mg/L (I=0.01mol/L and pH=7), it was shown that the colloid (1kDa-0.45μm) component of P accounted for 10.6%, 11.6%, 6.5%, and 4.0% of remaining total P concentration in the supernatant after P adsorption onto ferrihydrite (FH), goethite (GE), ferrihydrite-humic acid complex (FH-HA), goethite-humic acid complex (GE-HA), respectively. The oxide aggregates was the main mechanism for the formation of the colloid P in the supernatant. And colloidal adsorbent particles co-existing in the supernatant were another important reason for it. Additionally, dissolve organic matter dissolved from iron oxide-HA complexes could occupy large adsorption sites of colloidal iron causing less colloid P in the supernatant. Ultimately, we believe that the findings can provide a new way to deeply interpret the geochemical cycling of P, even when considering other contaminants such as organic pollutants, heavy metal ions, and arsenate at the sediment/soil-water interface in the real environment. Copyright © 2016. Published by Elsevier B.V.

Biological nutrients removal from the supernatant originating from the anaerobic digestion of the organic fraction of municipal solid waste.

Science.gov (United States)

Malamis, S; Katsou, E; Di Fabio, S; Bolzonella, D; Fatone, F

2014-09-01

This study critically evaluates the biological processes and techniques applied to remove nitrogen and phosphorus from the anaerobic supernatant produced from the treatment of the organic fraction of municipal solid waste (OFMSW) and from its co-digestion with other biodegradable organic waste (BOW) streams. The wide application of anaerobic digestion for the treatment of several organic waste streams results in the production of high quantities of anaerobic effluents. Such effluents are characterized by high nutrient content, because organic and particulate nitrogen and phosphorus are hydrolyzed in the anaerobic digestion process. Consequently, adequate post-treatment is required in order to comply with the existing land application and discharge legislation in the European Union countries. This may include physicochemical and biological processes, with the latter being more advantageous due to their lower cost. Nitrogen removal is accomplished through the conventional nitrification/denitrification, nitritation/denitritation and the complete autotrophic nitrogen removal process; the latter is accomplished by nitritation coupled with the anoxic ammonium oxidation process. As anaerobic digestion effluents are characterized by low COD/TKN ratio, conventional denitrification/nitrification is not an attractive option; short-cut nitrogen removal processes are more promising. Both suspended and attached growth processes have been employed to treat the anaerobic supernatant. Specifically, the sequencing batch reactor, the membrane bioreactor, the conventional activated sludge and the moving bed biofilm reactor processes have been investigated. Physicochemical phosphorus removal via struvite precipitation has been extensively examined. Enhanced biological phosphorus removal from the anaerobic supernatant can take place through the sequencing anaerobic/aerobic process. More recently, denitrifying phosphorus removal via nitrite or nitrate has been explored. The removal of

Grain dust induces IL-8 production from bronchial epithelial cells: effect on neutrophil recruitment.

Science.gov (United States)

Park, H S; Suh, J H; Kim, S S; Kwon, O J

2000-06-01

There have been several investigations suggesting an involvement of activated neutrophils in the development of grain dust (GD)-induced occupational asthma. Interleukin-8 in the sputa from GD-induced asthmatic patients increased significantly after the exposure to GD. To confirm IL-8 production from bronchial epithelial cells when exposed to GD, and to evaluate the role of IL-8 on neutrophil recruitment. We cultured Beas-2B, a bronchial epithelial cell line. To observe GD-induced responses, four different concentrations ranging from 1 to 200 microg/mL of GD were incubated for 24 hours and compared with those without incubation of GD. To evaluate the effect of pro-inflammatory cytokines on IL-8 production and neutrophil chemotaxis, epithelial cells were incubated with peripheral blood mononuclear cell (PBMC) culture supernatant derived from subjects with GD-induced asthma exposed to 10 microg/mL of GD, and then compared with those without addition of PBMC supernatant. The level of released IL-8 in the supernatant was measured by enzyme-linked immunosorbent assay. Neutrophil chemotactic activity of the culture supernatant was determined by modified Boyden chamber method. Interleukin-8 production and neutrophil chemotactic activity from bronchial epithelial cells significantly increased with additions of GD in a dose-dependent manner (P < .05, respectively), and were significantly augmented with additions of PBMC supernatant (P < .05, respectively) at each concentration. Close correlation was noted between neutrophil chemotactic activity and IL-8 level (r = 0.87, P < .05). Compared with the untreated sample, pre-treatment of anti-IL-8 antibody induced a significant suppression (up to 67.2%) of neutrophil chemotactic activity in a dose-dependent manner. These results suggest that IL-8 produced from bronchial epithelial cells may be a major cytokine, which induces neutrophil migration into the airways when exposed to GD.

Establishment of an inducing medium for type III effector secretion in Xanthomonas campestris pv. campestris

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Guo-Feng Jiang

2013-09-01

Full Text Available It is well known that the type III secretion system (T3SS and type III (T3 effectors are essential for the pathogenicity of most bacterial phytopathogens and that the expression of T3SS and T3 effectors is suppressed in rich media but induced in minimal media and plants. To facilitate in-depth studies on T3SS and T3 effectors, it is crucial to establish a medium for T3 effector expression and secretion. Xanthomonas campestris pv. campestris (Xcc is a model bacterium for studying plant-pathogen interactions. To date no medium for Xcc T3 effector secretion has been defined. Here, we compared four minimal media (MME, MMX, XVM2, and XOM2 which are reported for T3 expression induction in Xanthomonas spp. and found that MME is most efficient for expression and secretion of Xcc T3 effectors. By optimization of carbon and nitrogen sources and pH value based on MME, we established XCM1 medium, which is about 3 times stronger than MME for Xcc T3 effectors secretion. We further optimized the concentration of phosphate, calcium, and magnesium in XCM1 and found that XCM1 with a lower concentration of magnesium (renamed as XCM2 is about 10 times as efficient as XCM1 (meanwhile, about 30 times stronger than MME. Thus, we established an inducing medium XCM2 which is preferred for T3 effector secretion in Xcc.

Parenteral medium-chain triglyceride-induced neutrophil activation is not mediated by a Pertussis Toxin sensitive receptor.

Science.gov (United States)

Versleijen, Michelle W J; van Esterik, Joantine C J; Roelofs, Hennie M J; van Emst-de Vries, Sjenet E; Willems, Peter H G M; Wanten, Geert J A

2009-02-01

Lipid-induced immune modulation might contribute to the increased infection rate that is observed in patients using parenteral nutrition. We previously showed that emulsions containing medium-chain triglycerides (LCT/MCTs or pure MCTs), but not pure long-chain triglycerides (LCTs), impair neutrophil functions, modulate cell-signaling and induce neutrophil activation in vitro. It has recently been shown that medium-chain fatty acids are ligands for GPR84, a pertussis toxin (PT)-sensitive G-protein-coupled receptor (GPCR). This finding urged us to investigate whether MCT-induced neutrophil activation is mediated by PT-sensitive GPCRs. Neutrophils isolated from blood of healthy volunteers were pre-incubated with PT (0.5-1 microg/mL, 1.5 h) and analyzed for the effect of this pre-incubation on LCT/MCT (2.5 mmol/L)-dependent modulation of serum-treated zymosan (STZ)-induced intracellular Ca(2+) mobilization and on LCT/MCT (5 mmol/L)-induced expression of cell surface adhesion (CD11b) and degranulation (CD66b) markers and oxygen radical (ROS) production. PT did not inhibit the effects of LCT/MCT on the STZ-induced increase in cytosolic free Ca(2+) concentration. LCT/MCT increased ROS production to 146% of unstimulated cells. However, pre-incubation with PT did not inhibit the LCT/MCT-induced ROS production. Furthermore, the LCT/MCT-induced increase in CD11b and CD66b expression (196% and 235% of unstimulated cells, respectively) was not inhibited by pre-incubation with PT. LCT/MCT-induced neutrophil activation does not involve the action of a PT-sensitive G-protein-coupled receptor.

Morphology of primary human venous endothelial cell cultures before and after culture medium exchange.

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Krüger-Genge, A; Fuhrmann, R; Jung, F; Franke, R P

2015-01-01

The evaluation of the interaction of human, venous endothelial cells (HUVEC) with body foreign materials on the cellular level cannot be performed in vivo, but is investigated in vitro under standard culture conditions. To maintain the vitality, proliferation and morphology of HUVEC seeded on body foreign substrates over days, the cell culture medium is usually exchanged every second day. It is well known, that alterations in the microenvironment of cells bear the risk of influencing cell morphology and function. In the current study the influence of cell culture medium exchange on HUVEC cytoskeletal microfilament structure and function was investigated. HUVEC in the third passage were seeded on extracellular matrix (ECM) - which was secreted from bovine corneal endothelial cells on glass- until functional confluence was reached. The experiment started 11 days after HUVEC seeding with an exchange of the cell culture medium followed by a staining of the actin microfilaments with phalloidin-rhodamin 1.5 and 5 minutes after medium exchange. The microfilaments were documented by use of an Olympus microscope (IMT-2) equipped with a UV lamp and online connected to a TV chain (Sony XC 50 ST/monochrome) implying an OPTIMAS - Image analysis system. Prostacyclin was analysed in the cell culture supernatant. 1.5 min after culture medium exchange in the functionally confluent cultures a slight disturbance of the actin microfilament structure with a broadening of the marginal filament band, a partial disconnection of cell-cell contacts and the appearance of intercellular fenestrations were observed. 5 minutes after medium exchange a redevelopment of the slightly disturbed microfilament structure with a condensation and narrowing of the marginal filament band was seen. 12 h later a further consolidation of the microfilament structure occurred. In addition, a perturbation of the cultured HUVEC occurred after cell culture medium exchange. The prostacyclin concentration in the

Selection of the treatment method for the West Valley alkaline supernatant

International Nuclear Information System (INIS)

Carl, D.E.; Leonard, I.M.

1987-02-01

As part of the West Valley Demonstration Project (WVDP), th PUREX supernatant stored in Tank 8d-2 will be partially decontaminated before encapsulation in the final glass form. This report discusses selection of a method for removing Cs-137, the major radioactive ion in the supernatant. Methods considered were: (1) electrodialysis; (2) hyperfiltration; (3) precipitation with ferrocyanide, NaTPB, or PTA; (4) organic ion exchange using Cs-100 or a biologically derived media; (5) chelation using DeVoe/Holbein compostions; and (6) inorganic ion exchange using Durasil, natural zeolities, IE-95 or IE-96 media. Several different methods of using inorganic ion exchange media were also reviewed including (1) four columns with elution, and (2) two, three, or four columns without elution. After the careful evaluation of experimental data with all process constraints taken into account, the inorganic exchange media IE-96 (Linde Ionsiv IE-96 synthetic zeolite) was chosen for WVDP cesium recovery. IE-96 was chosen for the following reasons: high sorption rate, a decontamination factor (DF) over 1000, excellent exchange capacity at WVDP conditions, compatability with the glass formers used for borosilicate glass in direct melter feed applications, and a history of successful application in radio chemical seperation for waste streams. 34 refs., 29 figs., 27 tabs

Comparison of EBV DNA viral load in whole blood, plasma, B-cells and B-cell culture supernatant.

Science.gov (United States)

Ouedraogo, David Eric; Bollore, Karine; Viljoen, Johannes; Foulongne, Vincent; Reynes, Jacques; Cartron, Guillaume; Vendrell, Jean-Pierre; Van de Perre, Philippe; Tuaillon, Edouard

2014-05-01

Epstein-Barr virus (EBV) genome quantitation in whole blood is used widely for therapeutic monitoring of EBV-associated disorders in immunosuppressed individuals and in patients with EBV-associated lymphoma. However, the most appropriate biological material to be used for EBV DNA quantitation remains a subject of debate. This study compare the detection rate and levels of EBV DNA from whole blood, plasma, enriched B-cells, and B-cell short-term culture supernatant using quantitative real-time PCR. Samples were collected from 33 subjects with either HIV infection or B-cell lymphoma. Overall, EBV DNA was detected in 100% of enriched B-cell samples, in 82% of B-cell culture supernatants, in 57% of plasma, and 42% of whole blood samples. A significant correlation for EBV viral load was found between enriched B-cell and B-cell culture supernatant material (ρ = 0.92; P cells (ρ = -0.02; P = 0.89), whole blood and plasma (ρ = 0.24; P = 0.24), or enriched B-cells and plasma (ρ = 0.08; P = 0.77). Testing of enriched B-cells appeared to be the most sensitive method for detection of EBV DNA as well as for exploration of the cellular reservoir. Quantitation of EBV DNA in plasma and B-cell culture supernatant may be of interest to assess EBV reactivation dynamics and response to treatment as well as to decipher EBV host-pathogen interactions in various clinical scenarios. © 2013 Wiley Periodicals, Inc.

Levodopa-Induced Dyskinesia Is Related to Indirect Pathway Medium Spiny Neuron Excitotoxicity: A Hypothesis Based on an Unexpected Finding

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Svetlana A. Ivanova

2016-01-01

Full Text Available A serendipitous pharmacogenetic finding links the vulnerability to developing levodopa-induced dyskinesia to the age of onset of Huntington’s disease. Huntington’s disease is caused by a polyglutamate expansion of the protein huntingtin. Aberrant huntingtin is less capable of binding to a member of membrane-associated guanylate kinase family (MAGUKs: postsynaptic density- (PSD- 95. This leaves more PSD-95 available to stabilize NR2B subunit carrying NMDA receptors in the synaptic membrane. This results in increased excitotoxicity for which particularly striatal medium spiny neurons from the indirect extrapyramidal pathway are sensitive. In Parkinson’s disease the sensitivity for excitotoxicity is related to increased oxidative stress due to genetically determined abnormal metabolism of dopamine or related products. This probably also increases the sensitivity of medium spiny neurons for exogenous levodopa. Particularly the combination of increased oxidative stress due to aberrant dopamine metabolism, increased vulnerability to NMDA induced excitotoxicity, and the particular sensitivity of indirect pathway medium spiny neurons for this excitotoxicity may explain the observed increased prevalence of levodopa-induced dyskinesia.

Ultrastructural Alterations in Lepocinclis acus (Euglenophyta Induced by Medium with High Organic Matter Content

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Visitación T. Conforti

2017-11-01

Full Text Available Ultrastructural changes induced by exposure to excess of organic matter were studied in Lepocinclis acus (ex Euglena acus. The cells isolated from the Matanza River, Buenos Aires, Argentina, were grown in soil water medium (SWM. When transferred to medium enriched with Bacteriological Peptone OXOID®, marked body deformation and a significant shortening and widening of the cells was observed. These changes were unexpected in a species with quite rigid cells, a condition previously shown in studies of the pellicle fine structure. Transmission electron microscopy observations suggest that cellular deformation might be facilitated by an increase in strip number, whereas in the original strips normal ultrastructure was maintained. An increase in number and volume of paramylon grains and vacuoles, as well as the presence of membrane whorls in vacuoles was observed. The fine structure of organisms grown in medium with and without organic matter enrichment was compared, and the systematic and ecological importance of morphological changes triggered by cell deformation was discussed.

Antidiabetic activity of medium-polar extract from the leaves of Stevia rebaudiana Bert. (Bertoni) on alloxan-induced diabetic rats

Science.gov (United States)

Misra, Himanshu; Soni, Manish; Silawat, Narendra; Mehta, Darshana; Mehta, B. K.; Jain, D. C.

2011-01-01

Objective: To investigate the medicative effects of medium-polar (benzene:acetone, 1:1, v/v) extract of leaves from Stevia rebaudiana (family Asteraceae) on alloxan-induced diabetic rats. Materials and Methods: Diabetes was induced in adult albino Wistar rats by intraperitoneal (i.p.) injection of alloxan (180 mg/kg). Medium-polar extract was administered orally at daily dose of 200 and 400 mg/kg body wt. basis for 10 days. The control group received normal saline (0.9%) for the same duration. Glibenclamide was used as positive control reference drug against Stevia extract. Results: Medium-polar leaf extract of S. rebaudiana (200 and 400 mg/kg) produced a delayed but significant (P Stevia extract was found to antagonize the necrotic action of alloxan and thus had a re-vitalizing effect on β-cells of pancreas. PMID:21687353

Parenteral medium-chain triglyceride-induced neutrophil activation is not mediated by a Pertussis Toxin sensitive receptor.

NARCIS (Netherlands)

Versleijen, M.W.J.; Esterik, J.C. van; Schaap-Roelofs, H.M.J.; Emst-de Vries, S.E. van; Willems, P.H.G.M.; Wanten, G.J.A.

2009-01-01

BACKGROUND & AIMS: Lipid-induced immune modulation might contribute to the increased infection rate that is observed in patients using parenteral nutrition. We previously showed that emulsions containing medium-chain triglycerides (LCT/MCTs or pure MCTs), but not pure long-chain triglycerides

Leptospira interrogans activation of peripheral blood monocyte glycolipoprotein demonstrated in whole blood by the release of IL-6

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F. Dorigatti

2005-06-01

Full Text Available Glycolipoprotein (GLP from pathogenic serovars of Leptospira has been implicated in the pathogenesis of leptospirosis by its presence in tissues of experimental animals with leptospirosis, the inhibition of the Na,K-ATPase pump activity, and induced production of cytokines. The aims of the present study were to investigate the induction of IL-6 by GLP in peripheral blood mononuclear cells (PBMC and to demonstrate monocyte stimulation at the cellular level in whole blood from healthy volunteers. PBMC were stimulated with increasing concentrations (5 to 2500 ng/ml of GLP extracted from the pathogenic L. interrogans serovar Copenhageni, lipopolysaccharide (positive control or medium (negative control, and supernatants were collected after 6, 20/24, and 48 h, and kept at -80ºC until use. Whole blood was diluted 1:1 in RPMI medium and cultivated for 6 h, with medium, GLP and lipopolysaccharide as described above. Monensin was added after the first hour of culture. Supernatant cytokine levels from PBMC were measured by ELISA and intracellular IL-6 was detected in monocytes in whole blood cultures by flow-cytometry. Monocytes were identified in whole blood on the basis of forward versus side scatter parameters and positive reactions with CD45 and CD14 antibodies. GLP ( > or = 50 ng/ml-induced IL-6 levels in supernatants were detected after 6-h incubation, reaching a peak after 20/24 h. The percentage of monocytes staining for IL-6 increased with increasing GLP concentration. Thus, our findings show a GLP-induced cellular activation by demonstrating the ability of GLP to induce IL-6 and the occurrence of monocyte activation in whole blood at the cellular level.

Royal Society of Tropical Medicine and Hygiene meeting at Manson House, London, 14 December 1995. Enteropathogenic Escherichia coli--mucosal infection models.

Science.gov (United States)

Frankel, G; Phillips, A D; Hicks, S; Dougan, G

1996-01-01

The formation of attaching and effacing (A/E) lesions is central to the pathogenesis of enteropathogenic Escherichia coli (EPEC)-mediated disease in humans and Citrobacter rodentium-mediated transmissible colonic hyperplasia in mice. Closely related outer membrane proteins, known as intimins, are required for formation of the A/E lesion by both EPEC and C. rodentium. In this study we found similar ultrastructural damage in small intestinal biopsies from an EPEC-infected child and large bowel specimens from C. rodentium-infected mice. The C. rodentium-infected large bowel biopsies revealed massive hyperplastic reactions and the infected human small intestinal biopsies showed an increase in total crypt cell number and mitotic index. EPEC-infected small intestinal organ cultures revealed bacteria adhering in a localized pattern and evidence of A/E lesions. Covaspheres coated with a biologically active cell-binding domain of intimin also adhered to cells in a localized fashion but did not induce the characteristic A/E lesions.

Electrolytic destruction of oxalate ions in plutonium oxalate supernatant

International Nuclear Information System (INIS)

Michael, K.M.; Talnikar, S.G.; Jambunathan, U.; Kapoor, S.C.; Ramanujam, A.; Venkataraman, N.

1996-01-01

A simple and efficient electrolytic method is described for the destruction of the oxalate ions present in plutonium oxalate supernatant. Using platinum electrode and very little KMnO 4 , in situ generation of Mn 3+ ions is achieved which in turn destroys the oxalate. The use of lower current density helps in achieving maximum current efficiency. The end point is easily detectable by the pink colour of permanganate. By reversing the current, this slight excess of permanganate can be destroyed, thus avoiding the use of hydrogen peroxide. By this simple electrolytic method, the corrosive oxalate ion is completely destroyed and the salt content of the waste solution is considerably reduced. (author). 4 refs., 1 fig., 6 tabs

Effect of transplantation of olfactory ensheathing cell conditioned medium induced bone marrow stromal cells on rats with spinal cord injury

Science.gov (United States)

Feng, Linjie; Gan, Hongquan; Zhao, Wenguo; Liu, Yingjie

2017-01-01

Spinal cord injury is a serious threat to human health and various techniques have been deployed to ameliorate or cure its effects. Stem cells transplantation is one of the promising methods. The primary aim of the present study was to investigate the effect of the transplantation of olfactory ensheathing cell (OEC) conditioned medium-induced bone marrow stromal cells (BMSCs) on spinal cord injury. Rat spinal cord compression injury animal models were generated, and the rats divided into the following three groups: Group A, (control) Dulbecco's modified Eagle's medium-treated group; group B, normal BMSC-treated group; group C, OEC conditioned medium-induced BMSC-treated group. The animals were sacrificed at 2, 4 and 8 weeks following transplantation for hematoxylin and eosin staining, and fluorescence staining of neurofilament protein, growth associated protein-43 and neuron-specific nuclear protein. The cavity area of the spinal cord injury was significantly reduced at 2 and 4 weeks following transplantation in group C, and a significant difference between the Basso, Beattie and Bresnahan score in group C and groups A and B was observed. Regenerated nerve fibers were observed in groups B and C; however, a greater number of regenerated nerve fibers were observed in group C. BMSCs induced by OEC conditioned medium survived in vivo, significantly reduced the cavity area of spinal cord injury, promoted nerve fiber regeneration following spinal cord injury and facilitated recovery of motor function. The present study demonstrated a novel method to repair spinal cord injury by using induced BMSCs, with satisfactory results. PMID:28656221

Culture supernatants from V. cholerae O1 ElTor strains isolated from different geographic areas induce cell vacuolation and cytotoxicity Cepas de V. cholerae O1 biotipo ElTor aisladas de diferente origen geográfico inducen vacuolización celular y citotoxicidad

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Jorge E Vidal

2009-02-01

Full Text Available OBJECTIVE: To investigate whether the HlyA-induced vacuolating effect is produced by V. cholerae O1 ElTor strains isolated from different geographic origins, including Mexico. MATERIAL AND METHODS: Supernatant-induced haemolysis, vacuolating activity and cytotoxicity in Vero cells were recorded. PCR, RFLP analysis and molecular cloning were performed. RESULTS: All ElTor strains analyzed induced cellular vacuolation. Ribotype 2 strains isolates from the U.S. gulf coast yielded the highest titer of vacuolating activity. Eight of nine strains were haemolytic, while all strains were PCR positive for the hlyA gene. We cloned the hlyA gene from two ElTor strains, a toxigenic (2514-88, ctxAB+ and a non-toxigenic Mexican strain (CM 91-3, ctxAB-. Supernatant from those recombinant E. coli strains induced haemolysis, cell vacuolation and cytotoxicity. RFLP-PCR analysis revealed similarities in the hlyA gene from all strains tested. CONCLUSION: The HlyA-induced vacuolating effect is a widespread phenotype of epidemic V. cholerae O1 ElTor strains.OBJETIVO: Analizar el efecto vacuolizante de cepas de V. cholerae O1 ElTor aisladas de diferente origen geográfico, incluyendo México. MATERIAL Y MÉTODOS: Se realizaron pruebas de hemolisis, vacuolización y citotoxicidad en células Vero, así como PCR, análisis por RFLP y clonación molecular. RESULTADOS: Todas las cepas indujeron el efecto vacuolizante. Las cepas del ribotipo 2, aisladas de las costas del Golfo en Estados Unidos, presentaron títulos altos de vacuolización. El gen hlyA fue amplificado en las nueve cepas mediante PCR, aunque sólo ocho fueron hemolíticas. Se clonó el gen hlyA de una cepa toxigénica (2514-88, ctxAB+ y de una cepa no toxigénica aislada en México (CM 91-3, ctxAB-. El sobrenadante de las clonas recombinantes indujo hemólisis, efecto vacuolizante y citotoxicidad. El RFLP mostró alta similitud del gen hlyA de las cepas estudiadas. CONCLUSIÓN: El efecto vacuolizante es un

Sampling and analysis of high level waste tank supernatant: an overview

International Nuclear Information System (INIS)

Goergen, C.R.

1981-01-01

The Savannah River Plant routinely samples its high level radioactive waste tank supernatants for analysis of major components. These results are important in maintaining proper levels of corrosion inhibiters for protection of the tank walls. Because the tank ambient temperature is elevated, the sample is heated to 70 0 C prior to removing aliquots for use in a variety of analytical methods. Typical analyses include density, pH, OH - , NO 3 - , and NO 2 - , with occasional requests for Al(OH) 4 - , CO 3 /sup =/, PO 4 /sup =/, SO 4 /sup =/, and various radionuclides

Enteropathogenic Escherichia coli translocate Tir and form an intimin-Tir intimate attachment to red blood cell membranes.

Science.gov (United States)

Shaw, Robert K; Daniell, Sarah; Frankel, Gad; Knutton, Stuart

2002-05-01

Type III secretion allows bacteria to inject effector proteins into host cells. In enteropathogenic Escherichia coli (EPEC) the type III secreted protein, Tir, is translocated to the host-cell plasma membrane where it functions as a receptor for the bacterial adhesin intimin, leading to intimate bacterial attachment and "attaching and effacing" (A/E) lesion formation. To study EPEC type III secretion the interaction of EPEC with monolayers of red blood cells (RBCs) has been exploited and in a recent study [Shaw, R. K., Daniell, S., Ebel, F., Frankel, G. & Knutton, S. (2001 ). Cell Microbiol 3, 213-222] it was shown that EPEC induced haemolysis of RBCs and translocation of EspD, a putative pore-forming type III secreted protein in the RBC membrane. Here it is demonstrated that EPEC are able to translocate and correctly insert Tir into the RBC membrane and produce an intimin-Tir intimate bacterial attachment, identical to that seen in A/E lesions. Following translocation Tir did not undergo any change in apparent molecular mass or become tyrosine-phosphorylated and there was no focusing of RBC cytoskeletal actin beneath intimately adherent bacteria, and no pedestal formation. This study, employing an RBC model of infection, has demonstrated that Tir translocation can be separated from host-cell-mediated Tir modifications; the data show that the EPEC type III protein translocation apparatus is sufficient to deliver and correctly insert Tir into host-cell membranes independent of eukaryotic cell functions.

Brucella abortus-infected B cells induce osteoclastogenesis.

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Pesce Viglietti, Ayelén Ivana; Arriola Benitez, Paula Constanza; Giambartolomei, Guillermo Hernán; Delpino, María Victoria

2016-09-01

Brucella abortus is an intracellular bacterium that establishes lifelong infections in livestock and humans although the mechanisms of its chronicity are poorly understood. Activated B cells have long lifespan and B. abortus infection activates B cells. Our results indicate that the direct infection of B cells with B. abortus induced matrix metalloproteinase-9 (MMP-9), receptor activator for NF κB ligand (RANKL), tumor necrosis factor (TNF)-α and interleukin (IL)-6 secretion. In addition, supernatants from B. abortus-infected B cells induced bone marrow-derived monocytes to undergo osteoclastogenesis. Using osteoprotegerin, RANKL's decoy receptor, we determined that RANKL is involved in osteoclastogenesis induced by supernatants from B. abortus-infected B cells. The results presented here shed light on how the interactions of B. abortus with B cells may have a role in the pathogenesis of brucellar osteoarticular disease. Copyright © 2016 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

Influence of the mycelium growth conditions on the production of amylolytic, proteolytic and pectinolytic enzymes by Aspergillus niger C

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Fiedurek, J.; Ilczuk, Z.; Lobarzewski, J. (Uniwersytet Marii Curie-Sklodowskiej, Lublin (Poland). Inst. Mikrobiologii i Biochemii)

1989-01-01

Various nitrogen and carbon sources, as well as natural products, were examined as inducers of the production of amylases, proteases and pectinases by A. niger C. A. niger C grown on wheat bran extract medium provided culture supernatants with the highest enzymatic activities. Some culture conditions, e.g. pH, medium temperature and time period of cultivation, were optimalized to improve the growth and enzyme biosynthesis by A. niger C. (orig.).

Comparative genomics and transcriptomics of Escherichia coli isolates carrying virulence factors of both enteropathogenic and enterotoxigenic E. coli.

Science.gov (United States)

Hazen, Tracy H; Michalski, Jane; Luo, Qingwei; Shetty, Amol C; Daugherty, Sean C; Fleckenstein, James M; Rasko, David A

2017-06-14

Escherichia coli that are capable of causing human disease are often classified into pathogenic variants (pathovars) based on their virulence gene content. However, disease-associated hybrid E. coli, containing unique combinations of multiple canonical virulence factors have also been described. Such was the case of the E. coli O104:H4 outbreak in 2011, which caused significant morbidity and mortality. Among the pathovars of diarrheagenic E. coli that cause significant human disease are the enteropathogenic E. coli (EPEC) and enterotoxigenic E. coli (ETEC). In the current study we use comparative genomics, transcriptomics, and functional studies to characterize isolates that contain virulence factors of both EPEC and ETEC. Based on phylogenomic analysis, these hybrid isolates are more genomically-related to EPEC, but appear to have acquired ETEC virulence genes. Global transcriptional analysis using RNA sequencing, demonstrated that the EPEC and ETEC virulence genes of these hybrid isolates were differentially-expressed under virulence-inducing laboratory conditions, similar to reference isolates. Immunoblot assays further verified that the virulence gene products were produced and that the T3SS effector EspB of EPEC, and heat-labile toxin of ETEC were secreted. These findings document the existence and virulence potential of an E. coli pathovar hybrid that blurs the distinction between E. coli pathovars.

Fate of cyanobacteria in drinking water treatment plant lagoon supernatant and sludge.

Science.gov (United States)

Pestana, Carlos J; Reeve, Petra J; Sawade, Emma; Voldoire, Camille F; Newton, Kelly; Praptiwi, Radisti; Collingnon, Lea; Dreyfus, Jennifer; Hobson, Peter; Gaget, Virginie; Newcombe, Gayle

2016-09-15

In conventional water treatment processes, where the coagulation and flocculation steps are designed to remove particles from drinking water, cyanobacteria are also concentrated into the resultant sludge. As a consequence, cyanobacteria-laden sludge can act as a reservoir for metabolites such as taste and odour compounds and cyanotoxins. This can pose a significant risk to water quality where supernatant from the sludge treatment facility is returned to the inlet to the plant. In this study the complex processes that can take place in a sludge treatment lagoon were investigated. It was shown that cyanobacteria can proliferate in the conditions manifest in a sludge treatment lagoon, and that cyanobacteria can survive and produce metabolites for at least 10days in sludge. The major processes of metabolite release and degradation are very dependent on the physical, chemical and biological environment in the sludge treatment facility and it was not possible to accurately model the net effect. For the first time evidence is provided to suggest that there is a greater risk associated with recycling sludge supernatant than can be estimated from the raw water quality, as metabolite concentrations increased by up to 500% over several days after coagulation, attributed to increased metabolite production and/or cell proliferation in the sludge. Copyright © 2016 Elsevier B.V. All rights reserved.

Evaluation of the Antibody in Lymphocyte Supernatant Assay to Detect Active Tuberculosis.

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Margaretha Sariko

Full Text Available We aimed to evaluate the antibody in lymphocyte supernatant (ALS assay as a biomarker to diagnose tuberculosis among adults from Tanzania with and without HIV.Adults admitted with suspicion for tuberculosis had sputa obtained for GeneXpert MTB/RIF, acid-fast bacilli smear and mycobacterial culture; blood was obtained prior to treatment initiation and after 4 weeks. Adults hospitalized with non-infectious conditions served as controls. Peripheral blood mononuclear cells were cultured unstimulated for 72 hours. Anti-mycobacterial antibodies were measured from culture supernatants by ELISA, using BCG vaccine as the coating antigen. Median ALS responses were compared between cases and controls at baseline and between cases over time.Of 97 TB cases, 85 were microbiologically confirmed and 12 were clinically diagnosed. Median ALS responses from TB cases (0.366 OD from confirmed cases and 0.285 from clinical cases were higher compared to controls (0.085, p<0.001. ALS responses did not differ based on HIV status, CD4 count or sputum smear status. Over time, the median ALS values declined significantly (0.357 at baseline; 0.198 after 4-weeks, p<0.001.Robust ALS responses were mounted by patients with TB regardless of HIV status, CD4 count, or low sputum bacillary burden, potentially conferring a unique niche for this immunologic biomarker for TB.

Culture supernatants of oral cancer cells induce impaired IFN-α production of pDCs partly through the down-regulation of TLR-9 expression.

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Han, Nannan; Zhang, Zun; Jv, Houyu; Hu, Jingzhou; Ruan, Min; Zhang, Chenping

2018-06-05

The aim of the present study was to investigate whether tumor-derived supernatants down-regulate the immune function of plasmacytoid dendritic cells (pDCs) in oral cancer and the potential molecular mechanisms of this effect. Immunohistochemistry (IHC) and flow cytometry were used to detect tumor-infiltrating and peripheral blood pDCs. MTS and flow cytometry were employed to evaluate the immune response of CD4 + T cells. Real-time PCR and ELISA assays were used to identify TLR-7 and TLR-9 expression, IFN-α production and tumor-secreted soluble cytokines. The proportion of pDCs (0.121%±0.043%) was significantly higher in Oral squamous cell carcinoma (OSCC) samples than in normal tissue (0.023%±0.016%) (P = 0.021). TLR9 mRNA was significantly lower in tumor-infiltrating pDCs and positively correlated to low IFN-α production (r = 0.956; Poral cancer cells negatively regulated TLR9 mRNA expression and the subsequent IFN-α production of pDCs, which inhibited the immune response of CD4 + T cells. The neutralizing antibodies blocking assay showed that the specific inhibitory effect of pDC functionality was associated with the soluble fraction of the oral cancer environment, which is mainly mediated by IL-10 and TGF-β cooperation. Tumor-derived supernatants may impair the function of tumor-infiltrating pDCs, which subsequently decreases the immune response of CD4 + T cells in human oral cancer through TGF-β- and IL-10- dependent mechanisms. Careful manipulation of these impaired pDCs may help develop an important alternative immunotherapy for the treatment of oral cancer. Copyright © 2018 Elsevier Ltd. All rights reserved.

Chemically induced immunotoxicity in a medium-term multiorgan bioassay for carcinogenesis with Wistar rats

International Nuclear Information System (INIS)

Spinardi-Barbisan, Ana Lucia Tozzi; Kaneno, Ramon; Barbisan, Luis Fernando; Viana de Camargo, Joao Lauro; Rodrigues, Maria Aparecida Marchesan

2004-01-01

A variety of chemicals can adversely affect the immune system and influence tumor development. The modifying potential of chemical carcinogens on the lymphoid organs and cytokine production of rats submitted to a medium-term initiation-promotion bioassay for carcinogenesis was investigated. Male Wistar rats were sequentially initiated with N-nitrosodiethylamine (DEN), N-methyl-N-nitrosourea (MNU), N-butyl-N-(4hydroxybutyl)nitrosamine (BBN), dihydroxy-di-n-propylnitrosamine (DHPN), and 1,2-dimethylhydrazine (DMH) during 4 weeks. Two initiated groups received phenobarbital (PB) or 2-acetylaminofluorene (2-AAF) for 25 weeks and two noninitiated groups received only PB or 2-AAF. A nontreated group was used as control. Lymphohematopoietic organs, liver, kidneys, lung, intestines, and Zymbal's gland were removed for histological analysis. Interleukin (IL)-2, IL-12, interferon gamma (IFN-γ), tumor necrosis factor alpha (TNF-α), IL-10, and transforming growth factor beta1 (TGF-β1) levels were determined by ELISA in spleen cell culture supernatants. At the fourth week, exposure to the initiating carcinogens resulted in cell depletion of the thymus, spleen and bone marrow, and impairment of IL-2, IL-12, and IFN-γ production. However, at the 30th week, no important alterations were observed both in lymphoid organs and cytokine production in the different groups. The results indicate that the initiating carcinogens used in the present protocol exert toxic effects on the lymphoid organs and affect the production of cytokines at the initiation step of carcinogenesis. This early and reversible depression of the immune surveillance may contribute to the survival of initiated cells facilitating the development of future neoplasia

Neuronal activation by mucosal biopsy supernatants from irritable bowel syndrome patients is linked to visceral sensitivity

NARCIS (Netherlands)

Buhner, Sabine; Braak, Breg; Li, Qin; Kugler, Eva Maria; Klooker, Tamira; Wouters, Mira; Donovan, Jemma; Vignali, Sheila; Mazzuoli-Weber, Gemma; Grundy, David; Boeckxstaens, Guy; Schemann, Michael

2014-01-01

Based on the discomfort/pain threshold during rectal distension, irritable bowel syndrome (IBS) patients may be subtyped as normo- or hypersensitive. We previously showed that mucosal biopsy supernatants from IBS patients activated enteric and visceral afferent neurons. We tested the hypothesis that

Influence of viscosity of the medium on the disposition of carbon nanotubes anisotropic structures formation induced by electric field

International Nuclear Information System (INIS)

Yakovenko, O.S.; Matsuj, L.Yu.; Zhuravkov, O.V.; Vovchenko, L.D.

2014-01-01

To obtain carbon nanotubes (CNT)-polymer composites with anisotropic physical properties an electric field application can be used. This investigation considers factors of CNT anisotropic distribution formation induced by electric field and consideration is supported with experimental results where some factors were varied. In the article an influence of magnitude and type of electric field and time of processing by electric field on CNT anisotropic structures formation in polymer mediums of different viscosities (oil, epoxy resins) is investigated. The aim of this work was to examine the CNT structuration process induced by electric field in viscous mediums and to find out the most optimal conditions of preparation of polymer/carbon composite materials (CM) with specified distribution of carbon filler induced by electric field. Scoping on polymer/carbon CM structuration was conducted by optical microscopy method. It was found that the main factors during CNT network formation are the type and viscosity of polymer binder and applied electric field parameters. It was observed that for high viscous polymer CNT network formation is unfeasible even at high applied electric field strength. But also for low viscous medium at relatively low electric field strength the CNT network formation is complicated too. And it was seen from optical observation that a type of the polymer variation causes different response of network form under the same experimental conditions. These distinctions are considered in the article

Treatment of serum with supernatants from cultures of Candida albicans reduces its serum-dependent phagocytosis Tratamento de soro com sobrenadante de cultura de Candida albicans reduz a fagocitose soro-dependente

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Aderbal Antonio dos Santos

2002-01-01

Full Text Available Candida albicans is a potent activator of the complement system, and heat labile opsonins produced by activation of C3 (C3b and iC3b enhance phagocytosis of C. albicans mediated by complement receptors. In this study we treated mouse serum with supernatants from cultures of a protease producer strain of C. albicans and evaluated the ability of this serum to enhance phagocytosis of C. albicans. Cell-free supernatants from cultures of C. albicans were concentrated 5 fold and added to mouse serum for 30 min at 37ºC, before using this serum for opsonization of glutaraldehyde-fixed yeast cells. We observed that normal mouse serum increased about 3 fold the phagocytosis of C. albicans by mice peritoneal macrophages, whereas supernatant-treated serum did not increase phagocytosis. This effect of supernatants on serum was prevented by addition of pepstatin (5 µg/ ml; an inhibitor of C. albicans acid proteases to the medium. Serum treated with supernatants from cultures of a protease-deficient mutant of C. albicans also increased about 3 fold phagocytosis of the yeast. These results suggest that a protease produced by C. albicans causes proteolysis of serum opsonins, thereby reducing the phagocytosis of the yeast.Candida albicans é um potente ativador do sistema complemento, e opsoninas lábeis ao calor produzidas por ativação de C3 (C3b e iC3b aumentam a fagocitose de C. albicans mediada por receptores de complemento. Neste estudo, tratamos o soro de camundongo com sobrenadante de culturas de uma cepa de C. albicans produtora de proteases e avaliamos a capacidade deste soro reduzir a fagocitose de C. albicans. Sobrenadantes livres de células obtidos de cultura de C. albicans foram concentrados 5 vezes e adicionados ao soro de camundongo por 30 minutos a 37ºC, antes deste soro ser usado para opsonização de C. albicans na forma de levedura e fixadas em glutaraldeido. Nós observamos que soro normal aumentou 3 vezes a fagocitose de C. albicans por

Comparison of diet-induced thermogenesis of foods containing medium- versus long-chain triacylglycerols.

Science.gov (United States)

Kasai, Michio; Nosaka, Naohisa; Maki, Hideaki; Suzuki, Yoshie; Takeuchi, Hiroyuki; Aoyama, Toshiaki; Ohra, Atsushi; Harada, Youji; Okazaki, Mitsuko; Kondo, Kazuo

2002-12-01

The purpose of this study was to investigate the effect of 5-10 g of medium-chain triacylglycerols (MCT) on diet-induced thermogenesis in healthy humans. The study compared diet-induced thermogenesis after ingestion of test foods containing MCT and long-chain triacylglycerols (LCT), using a double-blind, crossover design. Eight male and eight female subjects participated in study 1 and study 2, respectively. In both studies, the LCT was a blend of rapeseed oil and soybean oil. In study 1, the liquid meals contained 10 g MCT (10M), a mixture of 5 g MCT and 5 g LCT (5M5L), and 10 g LCT (10L). In study 2, the subjects were given a meal (sandwich and clear soup) with the mayonnaise or margarine containing 5 g of MCT or LCT. Postprandial energy expenditure was measured by indirect calorimetry before and during the 6 h after ingestion of the test meals. Diet-induced thermogenesis was significantly greater after 5M5L and 10M Ingestion as compared to 10L ingestion. Ingestion of the mayonnaise or margarine containing 5 g MCT caused significantly larger diet-induced thermogenesis as compared to that of LCT. These results suggest that, in healthy humans, the intake of 5-10 g of MCT causes larger diet-induced thermogenesis than that of LCT, irrespective of the form of meal containing the MCT.

Rapid point-of-care testing for epidermal growth factor receptor gene mutations in patients with lung cancer using cell-free DNA from cytology specimen supernatants.

Science.gov (United States)

Asaka, Shiho; Yoshizawa, Akihiko; Saito, Kazusa; Kobayashi, Yukihiro; Yamamoto, Hiroshi; Negishi, Tatsuya; Nakata, Rie; Matsuda, Kazuyuki; Yamaguchi, Akemi; Honda, Takayuki

2018-06-01

Epidermal growth factor receptor (EGFR) mutations are associated with responses to EGFR tyrosine kinase inhibitors (EGFR-TKIs) in non-small-cell lung cancer (NSCLC). Our previous study revealed a rapid point-of-care system for detecting EGFR mutations. This system analyzes cell pellets from cytology specimens using droplet-polymerase chain reaction (d-PCR), and has a reaction time of 10 min. The present study aimed to validate the performance of the EGFR d-PCR assay using cell-free DNA (cfDNA) from supernatants obtained from cytology specimens. Assay results from cfDNA supernatant analyses were compared with those from cell pellets for 90 patients who were clinically diagnosed with, or suspected of having, lung cancer (80 bronchial lavage fluid samples, nine pleural effusion samples and one spinal fluid sample). EGFR mutations were identified in 12 and 15 cases using cfDNA supernatants and cell pellets, respectively. The concordance rates between cfDNA-supernatant and cell‑pellet assay results were 96.7% [kappa coefficient (K)=0.87], 98.9% (K=0.94), 98.9% (K=0.79) and 98.9% (K=0.79) for total EGFR mutations, L858R, E746_A750del and T790M, respectively. All 15 patients with EGFR mutation-positive results, as determined by EGFR d-PCR assay using cfDNA supernatants or cell pellets, also displayed positive results by conventional EGFR assays using tumor tissue or cytology specimens. Notably, EGFR mutations were even detected in five cfDNA supernatants for which the cytological diagnoses of the corresponding cell pellets were 'suspicious for malignancy', 'atypical' or 'negative for malignancy.' In conclusion, this rapid point-of-care system may be considered a promising novel screening method that may enable patients with NSCLC to receive EGFR-TKI therapy more rapidly, whilst also reserving cell pellets for additional morphological and molecular analyses.

Preirradiation of medium induces a subsequent stimulation or inhibition of growth according to the physiological state in Synechococcus lividus in culture

International Nuclear Information System (INIS)

Conter, A.

1987-01-01

The proliferation of Synechococcus lividus cells grown in preirradiated medium was compared with the proliferation of cells grown in a shielded or freshly prepared medium. Aging of medium in a shielded chamber resulted in a slight inhibiting effect on growth in every phase of the cell cycle which was used. Preirradiation of medium resulted in a stimulation of growth observed on Day 7 in cultures inoculated with cells selected in the deceleration phase and an inhibition of growth in cultures inoculated with exponentially growing cells. Addition of catalase (100 U X ml-1) counteracted the stimulating effect but did not modify the inhibiting effect induced by preirradiated medium. Results demonstrated the indirect effect of low doses of irradiation, implying the presence of hydrogen peroxide in radiostimulation and other radioproducts in the inhibitory effect

The effect of the external medium on the gravity-induced polarity of cytoplasmic streaming in Chara corallina (Characeae)

Science.gov (United States)

Staves, M. P.; Wayne, R.; Leopold, A. C.

1997-01-01

Gravity induces a polarity of cytoplasmic streaming in vertical internodal cells of Chara such that the downwardly directed stream moves faster than the upwardly directed stream. In order to determine whether the statolith theory (in which intracellular sedimenting particles are responsible for gravity sensing) or the gravitational pressure theory (in which the entire protoplast acts as the gravity sensor) best explain the gravity response in Chara internodal cells, we controlled the physical properties of the external medium, including density and osmolarity, with impermeant solutes and examined the effect on the polarity of cytoplasmic streaming. As the density of the external medium is increased, the polarity of cytoplasmic streaming decreases and finally disappears when the density of the external medium is equal to that of the cell (1015 kg/m3). A further increase in the density of the external medium causes a reversal of the gravity response. These results are consistent with the gravitational pressure theory of gravity sensing since the buoyancy of the protoplast is dependent on the difference between the density of the protoplast and the external medium, and are inconsistent with the statolith theory since the buoyancy of intracellular particles are unaffected by changes in the external medium.

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

OpenAIRE

Yao, Yongchang; Zhai, Zhichen; Wang, Yingjun

2014-01-01

Background. The ATDC5 cell line is regarded as an excellent cell model for chondrogenesis. In most studies with ATDC5 cells, insulin medium (IM) was used to induce chondrogenesis while chondrogenic medium (CM), which was usually applied in chondrogenesis of mesenchymal stem cells (MSCs), was rarely used for ATDC5 cells. This study was mainly designed to investigate the effect of IM, CM, and growth medium (GM) on chondrogenesis of ATDC5 cells. Methods. ATDC5 cells were, respectively, cultured ...

Biological Activities of Uric Acid in Infection Due to Enteropathogenic and Shiga-Toxigenic Escherichia coli

Science.gov (United States)

Broome, Jacqueline E.; Lis, Agnieszka

2016-01-01

In previous work, we identified xanthine oxidase (XO) as an important enzyme in the interaction between the host and enteropathogenic Escherichia coli (EPEC) and Shiga-toxigenic E. coli (STEC). Many of the biological effects of XO were due to the hydrogen peroxide produced by the enzyme. We wondered, however, if uric acid generated by XO also had biological effects in the gastrointestinal tract. Uric acid triggered inflammatory responses in the gut, including increased submucosal edema and release of extracellular DNA from host cells. While uric acid alone was unable to trigger a chloride secretory response in intestinal monolayers, it did potentiate the secretory response to cyclic AMP agonists. Uric acid crystals were formed in vivo in the lumen of the gut in response to EPEC and STEC infections. While trying to visualize uric acid crystals formed during EPEC and STEC infections, we noticed that uric acid crystals became enmeshed in the neutrophilic extracellular traps (NETs) produced from host cells in response to bacteria in cultured cell systems and in the intestine in vivo. Uric acid levels in the gut lumen increased in response to exogenous DNA, and these increases were enhanced by the actions of DNase I. Interestingly, addition of DNase I reduced the numbers of EPEC bacteria recovered after a 20-h infection and protected against EPEC-induced histologic damage. PMID:26787720

The doppler frequency shift caused by the inhomogeneities of a medium induced by pulses of intense laser radiation

Science.gov (United States)

Rozanov, N. N.; Kiselev, Al. S.; Kiselev, An. S.

2008-08-01

Self-reflection of pulses of intense laser radiation from an inhomogeneity induced by them in a medium with fast optical nonlinearity is analyzed. The reflected radiation is characterized by a considerable Doppler shift and by a signal magnitude that is sufficient for experimental detection.

Antidiabetic activity of medium-polar extract from the leaves of Stevia rebaudiana Bert. (Bertoni on alloxan-induced diabetic rats

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Himanshu Misra

2011-01-01

Full Text Available Objective: To investigate the medicative effects of medium-polar (benzene:acetone, 1:1, v/v extract of leaves from Stevia rebaudiana (family Asteraceae on alloxan-induced diabetic rats. Materials and Methods: Diabetes was induced in adult albino Wistar rats by intraperitoneal (i.p. injection of alloxan (180 mg/kg. Medium-polar extract was administered orally at daily dose of 200 and 400 mg/kg body wt. basis for 10 days. The control group received normal saline (0.9% for the same duration. Glibenclamide was used as positive control reference drug against Stevia extract. Results: Medium-polar leaf extract of S. rebaudiana (200 and 400 mg/kg produced a delayed but significant (P < 0.01 decrease in the blood glucose level, without producing condition of hypoglycemia after treatment, together with lesser loss in the body weight as compared with standard positive control drug glibenclamide. Conclusions: Treatment of diabetes with sulfonylurea drugs (glibenclamide causes hypoglycemia followed by greater reduction in body weight, which are the most worrisome effects of these drugs. Stevia extract was found to antagonize the necrotic action of alloxan and thus had a re-vitalizing effect on β-cells of pancreas.

The study of the peptide composition of the supernatants from mealworm Tenebrio molitor larvae and goldfish Carassius auratus during cold acclimation

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А. К. Гулевский

2016-11-01

Full Text Available The molecular-mass distribution of peptides from supernatants, obtained from the tissues of larvae Tenebrio molitor and goldfish Carassius auratus during cold acclimation, has been determined by chromatography. The results showed that peptide spectrum of the supernatants from larvae T. molitor and C. auratus varied during cold acclimation. The supernatants from non-acclimated larvae of T. molitor and deacclimated fish possessed the highest number of peptide fractions. Furthermore, the cold-acclimated larvae of T. molitor had the peptide fractions of the low molecular weight (ca. 5.4×102 ÷22.6×102 Da, and non-acclimated insects had the peptides of the high molecular weight (ca. 46.8×102÷66×102 Da. Next, the organ-specific changes of the peptide composition of the goldfish during winter deacclimation have been revealed. Specifically, the low molecular weight peptides (ca. (14.1 ± 0.3×102 and (6.75 ± 0.25×102 Da, have been detected in the C. auratus muscles, and both the high (ca. (67.83 ± 0.21×102 ( ca. 64.16 ± 0.26×102 Da and low (ca. (34.1 ± 1.0×102 and (14.29 ± 0.15×102 Da molecular weight peptides have been detected in the liver. Quantitative and qualitative changes in the peptide spectra from supernatants of the T. molitor and C. auratus during cold acclimation could be one of the mechanisms of their natural adaptation to low temperatures.

Comparative analysis of virulence determinants, antibiotic susceptibility patterns and serogrouping of atypical enteropathogenic Escherichia coli versus typical enteropathogenic E. coli in India.

Science.gov (United States)

Malvi, Supriya; Appannanavar, Suma; Mohan, Balvinder; Kaur, Harsimran; Gautam, Neha; Bharti, Bhavneet; Kumar, Yashwant; Taneja, Neelam

2015-10-01

The epidemiology of enteropathogenic Escherichia coli (EPEC) and the significance of isolation of atypical EPEC (aEPEC) in childhood diarrhoea have not been well studied in an Indian context. A comparative study was undertaken to investigate virulence determinants, antibiotic susceptibility patterns and serogrouping of typical EPEC (tEPEC) versus aEPEC causing diarrhoea in children. A total of 400 prospective and 500 retrospective E. coli isolates were included. PCR was performed for eae, bfpA, efa, nleB, nleE, cdt, ehxA and paa genes. The Clinical and Laboratory Standards Institute's disc diffusion test was used to determine the antimicrobial susceptibility. Phenotypic screening of extended spectrum β-lactamases (ESBLs), AmpC and Klebsiella pneumoniae carbapenemase (KPC) production, and molecular detection of bla(NDM-1), bla(VIM), bla(CTX-M-15), bla(IMP) and bla(KPC) were performed. aEPEC (57.6 %) were more common as compared with tEPEC (42.3 %). The occurrence of virulence genes was observed to be three times higher in aEPEC as compared with tEPEC, efa1 (14.7 % of aEPEC, 4 % of tEPEC) being the most common. Most of the isolates did not belong to the classical EPEC O-serogroups. The highest resistance was observed against amoxicillin (93.22 %) followed by quinolones (83 %), cephalosporins (37.28 %), cotrimoxazole (35.59 %) and carbapenems (30.5 %). Overall equal numbers of aEPEC (41.17 %) and tEPEC (40 %) were observed to be multidrug-resistant. Fifteen EPEC strains demonstrated presence of ESBLs, five produced AmpC and four each produced metallo-β-lactamases and KPC-type carbapenemases; eight, seven and one isolate(s) each were positive for bla(VIM), bla(CTX-M-15) and bla(NDM-1), respectively. Here, to the best of our knowledge, we report for the first time on carbapenem resistance and the presence of bla(NDM-1) and bla(CTX-M-15) in EPEC isolates from India.

Phosphate removal from digested sludge supernatant using modified fly ash.

Science.gov (United States)

Xu, Ke; Deng, Tong; Liu, Juntan; Peng, Weigong

2012-05-01

The removal of phosphate in digested sludge supernatant by modified coal fly ash was investigated in this study. Modification of the fly ash by the addition of sulfuric acid could significantly enhance its immobilization ability. The experimental results also showed that adsorption of phosphate by the modified fly ash was rapid with the removal percentage of phosphate reaching an equilibrium of 98.62% in less than 5 minutes. The optimum pH for phosphate removal was 9 and the removal percentage increased with increasing adsorbent dosage. The effect of temperature on phosphate removal efficiency was not significant from 20 to 40 degrees C. X-ray diffraction and scanning electron microscope analyses showed that phosphate formed an amorphous precipitate with water-soluble calcium, aluminum, and iron ions in the modified fly ash.

Interference between initial and final state radiation in a QCD medium

CERN Document Server

Armesto, Nestor; Martinez, Mauricio; Mehtar-Tani, Yacine; Salgado, Carlos A.

2012-01-01

We investigate the color coherence pattern between initial and final state radiation in the presence of a QCD medium. We derive the medium-induced gluon spectrum of an "asymptotic" parton which suffers a hard scattering and subsequently crosses the medium. The angular distribution of the induced gluon spectrum is modified when one includes interference terms between the incoming and the outgoing parton at finite angle between them. The coherent, incoherent and soft limits of the medium-induced gluon spectrum are studied. In the soft limit, we provide a simple and intuitive probabilistic picture which could be of interest for Monte Carlo implementations. The configuration studied here may have phenomenological consequences in high energy nuclear collisions.

Thrombomodulin and von Willebrand factor as markers of radiation-induced endothelial injury

International Nuclear Information System (INIS)

Zhou Quansheng; Zhao Yimin; Li Peixia; Bai Xia; Ruan Changgeng

1992-02-01

Cultured confluent human umbilical vein endothelial cells were irradiated in vitro by 60 Co-gamma ray at doses from 0 to 50 Gy. After irradiation Thrombomodulin in the supernatants of endothelial cell culture medium, on the surface of the cells and within the cells was measured at different times over six days. At twenty-four hours after irradiation, an increase in the release of Thrombomodulin and von Willebrand factor from irradiated endothelial cells and an increase in the number of molecules and the activity of Thrombomodulin on the surface of the cells were observed, which were radiation-dose dependent. The capacity of the cells to produce and release Thrombomodulin was decreased from two to six days after exposure to 60 Co-gamma ray. Our data indicate that radiation can injure endothelial cells and that Thrombomodulin may be as a marker of radiation-induced endothelial cell injury. The relationship between dysfunction of irradiated endothelial cells and the pathological mechanisms of acute radiation sickness are discussed

Proteomic profiling in incubation medium of mouse, rat and human precision-cut liver slices for biomarker detection regarding acute drug-induced liver injury

NARCIS (Netherlands)

van Swelm, Rachel P L; Hadi, Mackenzie; Laarakkers, Coby M M; Masereeuw, R.|info:eu-repo/dai/nl/155644033; Groothuis, Geny M M; Russel, Frans G M

Drug-induced liver injury is one of the leading causes of drug withdrawal from the market. In this study, we investigated the applicability of protein profiling of the incubation medium of human, mouse and rat precision-cut liver slices (PCLS) exposed to liver injury-inducing drugs for biomarker

Biological synthesis of very small silver nanoparticles by culture supernatant of Klebsiella pneumonia: The effects of visible-light irradiation and the liquid mixing process

International Nuclear Information System (INIS)

Mokhtari, Narges; Daneshpajouh, Shahram; Seyedbagheri, Seyedali; Atashdehghan, Reza; Abdi, Khosro; Sarkar, Saeed; Minaian, Sara; Shahverdi, Hamid Reza; Shahverdi, Ahmad Reza

2009-01-01

This study has investigated different visible-light irradiation's effect on the formation of silver nanoparticles from silver nitrate using the culture supernatant of Klebsiella pneumonia. Our study shows that visible-light emission can significantly prompt the synthesis of silver nanoparticles. Also, the study experimentally investigated the liquid mixing process effect on silver nanoparticle synthesis by visible-light irradiation. This study successfully synthesized uniformly dispersed silver nanoparticles with a uniform size and shape in the range of 1-6 nm with an average size of 3 nm. Furthermore, the study investigated the mechanism of the reduction of silver ions by culture supernatant of K. pneumonia, and used X-ray diffraction to characterize silver chloride as an intermediate compound. Silver chloride was prepared synthetically and used as a substrate for the synthesis of silver nanoparticles by culture supernatant of K. pneumonia. The silver nanoparticles have been prepared from silver chloride during this investigation for the first time.

Biological synthesis of very small silver nanoparticles by culture supernatant of Klebsiella pneumonia: The effects of visible-light irradiation and the liquid mixing process

Energy Technology Data Exchange (ETDEWEB)

Mokhtari, Narges [Department of Pharmaceutical Biotechnology and Biotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of); Daneshpajouh, Shahram; Seyedbagheri, Seyedali; Atashdehghan, Reza [Hydrometallurgy Research Unit, Research and Development Center, National Iranian Copper Industries Company, Sarcheshmeh, Rafsanjan (Iran, Islamic Republic of); Abdi, Khosro [Department of Pharmaceutical Biotechnology and Biotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of); Sarkar, Saeed [Research Center for Science and Technology in Medicine, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of); Minaian, Sara [Department of Pharmaceutical Biotechnology and Biotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of); Shahverdi, Hamid Reza [Department of Material Science, Faculty of Engineering, Tarbiat Modares University, Tehran (Iran, Islamic Republic of); Shahverdi, Ahmad Reza, E-mail: shahverd@sina.tums.ac.ir [Department of Pharmaceutical Biotechnology and Biotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of)

2009-06-03

This study has investigated different visible-light irradiation's effect on the formation of silver nanoparticles from silver nitrate using the culture supernatant of Klebsiella pneumonia. Our study shows that visible-light emission can significantly prompt the synthesis of silver nanoparticles. Also, the study experimentally investigated the liquid mixing process effect on silver nanoparticle synthesis by visible-light irradiation. This study successfully synthesized uniformly dispersed silver nanoparticles with a uniform size and shape in the range of 1-6 nm with an average size of 3 nm. Furthermore, the study investigated the mechanism of the reduction of silver ions by culture supernatant of K. pneumonia, and used X-ray diffraction to characterize silver chloride as an intermediate compound. Silver chloride was prepared synthetically and used as a substrate for the synthesis of silver nanoparticles by culture supernatant of K. pneumonia. The silver nanoparticles have been prepared from silver chloride during this investigation for the first time.

Enhancement in irradiated mononuclear cells in culture of mitogen-induced incorporation of [3H]thymidine by homologous conditioned medium

International Nuclear Information System (INIS)

Sandru, G.; Greiner, R.

1994-01-01

Incorporation of [ 3 H]thymidine in irradiated peripheral blood mononuclear cell cultures irradiated in vitro was stimulated significantly by either concanavalin A or phytohemagglutinin only in the presence of homologous conditioned medium. Production of this activity by mononuclear cells was enhanced by irradiation and/or pulsed exposure to puromycin but was abolished by actinomycin D. Addition of anti-interleukin 1 or anti-interleukin 2 monoclonal antibodies to the conditioned medium before assay did not influence the stimulatory action. A similar significant stimulation of mononuclear cell cultures irradiated with 6 Gy by concanavalin A was obtained when purified preparations of homologous conditioned medium were used in the assay. Purification was done by ultrafiltration and concentration, heparin agarose chromatography, ammonium sulfate precipitation, concanavalin A agarose chromatography, DEAE-ion exchange chromatography and HPLC gel filtration chromatography. With SDS-PAGE and silver staining, the active HPLC fraction gave one band of 50 kDa, suggesting that this protein is responsible for the co-stimulatory effect of homologous conditioned medium for both mitogen-induced irradiated and nonirradiated mononuclear cell cultures. 42 refs., 9 figs., 3 tabs

Food waste co-digestion with slaughterhouse waste and sewage sludge: Digestate conditioning and supernatant quality.

Science.gov (United States)

Borowski, Sebastian; Boniecki, Paweł; Kubacki, Przemysław; Czyżowska, Agata

2018-04-01

In this study, the anaerobic mesophilic co-digestion of food waste (FW) with municipal sewage sludge (MSS) and slaughterhouse waste (SHW) was undertaken in 3-dm 3 laboratory reactors as well as in 50-dm 3 reactors operated in semi-continuous conditions. The highest methane yield of around 0.63 m 3 CH 4 /kgVS fed was achieved for the mixture of FW and SHW treated in the laboratory digester operated at solids retention time (SRT) of 30 days, whereas the co-digestion of FW with MSS under similar operating conditions produced 0.46 m 3 of methane from 1 kgVS fed . No significant differences between methane yields from laboratory digesters and large-scale reactors were reported. The conditioning tests with the digestates from reactor experiments revealed the highest efficiency of inorganic coagulants among all investigated chemicals, which applied in a dose of 10 g/kg allowed to reduce capiliary suction time (CST) of the digestate below 20 s. The combined conditioning with coagulants and bentonite did not further reduce the CST value but improved the quality of the digestate supernatant. In particular, the concentrations of suspended solids, COD as well as metals in the supernatant were considerably lowered. Copyright © 2017. Published by Elsevier Ltd.

ALTERED EXPRESSION OF SURFACE RECEPTORS AT EA.HY926 ENDOTHELIAL CELL LINE INDUCED WITH PLACENTAL SECRETORY FACTORS

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O. I. Stepanova

2012-01-01

Full Text Available Abstract. Placental cell populations produce a great variety of angiogenic factors and cytokines than control angiogenesis in placenta. Functional regulation of endothelial cells proceeds via modulation of endothelial cell receptors for endogenous angiogenic and apoptotic signals. Endothelial phenotype alteration during normal pregnancy and in cases of preclampsia is not well understood. The goal of this investigation was to evaluate altered expression of angiogenic and cytokine receptors at EA.hy926 endothelial cells under the influence of placental tissue supernatants. Normal placental tissue supernatants from 1st and 3rd trimesters, and pre-eclamptic placental tissue supernatants (3rd trimester stimulated angiogenic and cytokine receptors expression by the cultured endothelial cells, as compared with their background expression. Tissue supernatants from placental samples of 3rd trimester caused a decreased expression of angiogenic and cytokine receptors by endothelial cells, thus reflecting maturation of placental vascular system at these terms. Supernatants from preeclamptic placental tissue induced an increase of CD119 expression, in comparison with normal placental supernatants from the 3rd trimester. This finding suggests that IFNγ may be a factor of endothelial activation in pre-eclampsia. The study was supported by grants ГК №02.740.11.0711, НШ-3594.2010.7., and МД-150.2011.7.

Extracellular Protease Activity of Enteropathogenic Escherechia coli on Mucin Substrate

Directory of Open Access Journals (Sweden)

SRI BUDIARTI

2007-03-01

Full Text Available Enteropathogenic Escherichia coli (EPEC causes gastrointestinal infections in human. EPEC invasion was initiated by attachment and aggressive colonization on intestinal surface. Attachment of EPEC alter the intestine mucosal cells. Despite this, the pathogenic mechanism of EPEC infectior has not been fully understood. This research hypothesizes that extracellular proteolytic enzymes is necessary for EPEC colonization. The enzyme is secreted into gastrointestinal milieu and presumably destroy mucus layer cover the gastrointestinal tract. The objective of this study was to assay EPEC extracellular protease enzyme by using mucin substrate. The activity of EPEC extracellular proteolytic enzyme on 1% mucin substrate was investigated. Non-pathogenic E. coli was used as a negative control. Positive and tentative controls were Yersinia enterocolitica and Salmonella. Ten EPEC strains were assayed, seven of them were able to degrade mucin, and the highest activity was produced by K1.1 strain. Both positive and tentative controls also showed the ability to digest 0.20% mucin.

Immunoproteomic analysis of antibody in lymphocyte supernatant in patients with typhoid fever in Bangladesh.

Science.gov (United States)

Charles, Richelle C; Liang, Li; Khanam, Farhana; Sayeed, M Abu; Hung, Chris; Leung, Daniel T; Baker, Stephen; Ludwig, Albrecht; Harris, Jason B; Larocque, Regina C; Calderwood, Stephen B; Qadri, Firdausi; Felgner, Philip L; Ryan, Edward T

2014-03-01

We have previously shown that an assay based on detection of anti-Salmonella enterica serotype Typhi antibodies in supernatant of lymphocytes harvested from patients presenting with typhoid fever (antibody in lymphocyte supernatant [ALS] assay) can identify 100% of patients with blood culture-confirmed typhoid fever in Bangladesh. In order to define immunodominant proteins within the S. Typhi membrane preparation used as antigen in these prior studies and to identify potential biomarkers unique to S. Typhi bacteremic patients, we probed microarrays containing 2,724 S. Typhi proteins with ALS collected at the time of clinical presentation from 10 Bangladeshis with acute typhoid fever. We identified 62 immunoreactive antigens when evaluating both the IgG and IgA responses. Immune responses to 10 of these antigens discriminated between individuals with acute typhoid infection and healthy control individuals from areas where typhoid infection is endemic, as well as Bangladeshi patients presenting with fever who were subsequently confirmed to have a nontyphoid illness. Using an ALS enzyme-linked immunosorbent assay (ELISA) format and purified antigen, we then confirmed that immune responses against the antigen with the highest immunoreactivity (hemolysin E [HlyE]) correctly identified individuals with acute typhoid or paratyphoid fever in Dhaka, Bangladesh. These observations suggest that purified antigens could be used with ALS and corresponding acute-phase activated B lymphocytes in diagnostic platforms to identify acutely infected patients, even in areas where enteric fever is endemic.

Enteropathogenic Escherichia coli: foe or innocent bystander?

Science.gov (United States)

Hu, Jia; Torres, Alfredo G.

2015-01-01

Enteropathogenic Escherichia coli (EPEC) remain one the most important pathogens infecting children and they are one of the main causes of persistent diarrhea worldwide. Historically, typical EPEC (tEPEC), defined as those isolates with the attaching and effacement (A/E) genotype (eae+), which possess bfpA+ and lack the stx- genes are found strongly associated with diarrheal cases. However, occurrence of atypical EPEC (aEPEC; eae+ bfpA- stx-) in diarrheal and asymptomatic hosts has made investigators question the role of these pathogens in human disease. Current epidemiological data is helping answering the question whether EPEC is mainly a foe or an innocent bystander during infection. PMID:25726041

Recovery of urinary nanovesicles from ultracentrifugation supernatants.

Science.gov (United States)

Musante, Luca; Saraswat, Mayank; Ravidà, Alessandra; Byrne, Barry; Holthofer, Harry

2013-06-01

Urinary vesicles represent a newly established source of biological material, widely considered to faithfully represent pathological events in the kidneys and the urogenital epithelium. The majority of currently applied isolation protocols involve cumbersome centrifugation steps to enrich vesicles from urine. To date, the efficiency of these approaches has not been investigated with respect to performing quantitative and qualitative analyses of vesicle populations in the pellet and supernatant (SN) fractions. After the series of differential centrifugations, the final SN was reduced to one-twentieth of the original volume by ammonium sulphate precipitation, with the precipitate pellet subjected to another round of differential centrifugations. Electron microscopy, dynamic light scattering and western blot analysis were used to characterize the vesicles present in individual fractions of interest. Pellets obtained after the second set of centrifugations at 200 000 g revealed the presence of vesicles which share a common marker profile, but with distinct differences from those seen in the initial 200 000 g pellet used as the reference. This suggests an enrichment of previously uncharacterized urinary vesicles still in solution after the initial centrifugation steps. Analysis of protein yields recovered post-ultracentrifugation revealed an additional 40% of vesicles retained from the SN. Moreover, these structures showed a formidable resistance to harsh treatments (e.g. 95% ammonium sulphate saturation, hypotonic dialysis, 0.3 M sodium hydroxide). Methods which employ differential centrifugations of native urine are remarkably ineffective and may lose a substantial population of biologically important vesicle species.

Recovery of plutonium from oxalate supernatant using 1-phenyl-3-methyl-4-benzoyl-5-pyrazolone

International Nuclear Information System (INIS)

Mohapatra, P.K.; Manchanda, V.K.; Gupta, K.K.; Singh, R.K.

1997-01-01

Extraction of Pu(IV) from oxalate supernatant was carried out employing varying concentrations of 1-phenyl-3-methyl-4-benzoyl-5-pyrazolone (PMBP). Near quantitative extraction of Pu(IV) from an aqueous solution of 0.2M oxalic acid and 3M HNO 3 was possible employing 0.05M PMBP solution in xylene. Extraction studies at different uranium loading conditions were carried out and conditions for quantitative stripping were arrived at. (author). 2 refs., 4 tabs

Medium modifications of vector mesons

International Nuclear Information System (INIS)

Pant, L.M.

2004-01-01

The omega photoproduction in nuclear medium with the ELSA facility at Bonn and the present status of the HADES collaboration to investigate the in-medium hadron properties in proton, heavy ions and hadron induced reactions at GSI, Darmstadt are presented. Efforts are under way to utilise the electron beam at Indore for experimental hadron physics in order to step into the intermediate energy nuclear physics regime. The skeletal outline of the high energy electron beam now available at CAT, Indore is discussed

Grain dust induces IL-8 production from bronchial epithelial cells: the effect of dexamethasone on IL-8 production.

Science.gov (United States)

Park, H S; Suh, J H; Kim, H Y; Kwon, O J; Choi, D C

1999-04-01

Recent publications have suggested an active participation of neutrophils to induce bronchoconstriction after inhalation of grain dust (GD). To further understand the role of neutrophils in the pathogenesis of GD-induced asthma, this investigation was designed to determine whether human bronchial epithelial cells could produce IL-8 production and to observe the effect of dexamethasone on IL-8 production. We cultured Beas-2B, a bronchial epithelial cell line. To observe GD-induced responses, four concentrations (1 to 200 microg/mL) of GD were incubated for 24 hours and compared with those without incubation of GD. To evaluate the effect of pro-inflammatory cytokines on IL-8 production, epithelial cells were incubated with peripheral blood mononuclear cell (PBMC) culture supernatant, which was derived from the culture of PBMC from a GD-induced asthmatic subject under the exposure to 10 microg/mL of GD, and compared with those cultured without addition of PBMC supernatant. The level of released IL-8 in the supernatant was measured by enzyme-linked immunosorbent assay. To evaluate the effect of dexamethasone on IL-8 production, four concentrations (5 to 5000 ng/mL) of dexamethasone were pre-incubated for 24 hours and the same experiments were repeated. There was significant production of IL-8 from bronchial epithelial cells with additions of GD in a dose-dependent manner (P < .05), which was significantly augmented with additions of PBMC supernatant (P < .05) at each concentration. Compared with the untreated sample, pretreatment of dexamethasone could induced a remarkable inhibitions (15% to 55%) of IL-8 production from bronchial epithelial cells in a dose-dependent manner. These results suggest that IL-8 production from bronchial epithelial cells may contribute to neutrophil recruitment occurring in GD-induced airway inflammation. The downregulation of IL-8 production by dexamethasone from bronchial epithelial cells may contribute to the efficacy of this compound in

Optically induced dielectropheresis sorting with automated medium exchange in an integrated optofluidic device resulting in higher cell viability.

Science.gov (United States)

Lee, Gwo-Bin; Wu, Huan-Chun; Yang, Po-Fu; Mai, John D

2014-08-07

We demonstrated the integration of a microfluidic device with an optically induced dielectrophoresis (ODEP) device such that the critical medium replacement process was performed automatically and the cells could be subsequently manipulated by using digitally projected optical images. ODEP has been demonstrated to generate sufficient forces for manipulating particles/cells by projecting a light pattern onto photoconductive materials which creates virtual electrodes. The production of the ODEP force usually requires a medium that has a suitable electrical conductivity and an appropriate dielectric constant. Therefore, a 0.2 M sucrose solution is commonly used. However, this requires a complicated medium replacement process before one is able to manipulate cells. Furthermore, the 0.2 M sucrose solution is not suitable for the long-term viability of cells. In comparison to conventional manual processes, our automated medium replacement process only took 25 minutes. Experimental data showed that there was up to a 96.2% recovery rate for the manipulated cells. More importantly, the survival rate of the cells was greatly enhanced due to this faster automated process. This newly developed microfluidic chip provided a promising platform for the rapid replacement of the cell medium and this was also the first time that an ODEP device was integrated with other active flow control components in a microfluidic device. By improving cell viability after cell manipulation, this design may contribute to the practical integration of ODEP modules into other lab-on-a-chip devices and biomedical applications in the future.

Characterization of culture supernatant proteins from Brucella abortus and its protection effects against murine brucellosis.

Science.gov (United States)

Lee, Jin Ju; Lim, Jeong Ju; Kim, Dae Geun; Simborio, Hannah Leah; Kim, Dong Hyeok; Reyes, Alisha Wehdnesday Bernardo; Min, WonGi; Lee, Hu Jang; Kim, Dong Hee; Chang, Hong Hee; Kim, Suk

2014-09-01

In this study, we characterized the secreted proteins of Brucella abortus into the enriched media under the bacterial laboratory growth condition and investigated the pathogenic importance of culture supernatant (CS) proteins to B. abortus infection. CS proteins from stationary phase were concentrated and analyzed using 2D electrophoresis. In MALDI TOF/TOF analysis, more than 27 proteins including CuZn SOD, Dps, Tat, OMPs, Adh, LivF, Tuf, SucC, GroEL and DnaK were identified. Cytotoxic effects of CS proteins were found to increase in a dose-dependent manner in RAW 264.7 cells. Upon B. abortus challenge into phagocytes, however, CS proteins pre-treated cells exhibited lower bacterial uptake and intracellular replication compared to untreated cells. Immunization with CS proteins induced a strong humoral and cell mediated immune responses and exhibited significant higher degree of protection against virulence of B. abortus infection compared to mice immunized with Brucella broth protein (BBP). Taken together, these results indicate that B. abortus secreted a number of soluble immunogenic proteins under laboratory culture condition, which can promote antibody production resulted in enhancing host defense against to subsequently bacterial infection. Moreover, further analysis of CS proteins may help to understand the pathogenic mechanism of B. abortus infection and host-pathogen interaction. Copyright © 2014 Elsevier Ltd. All rights reserved.

Ion exchange flowsheet for recovery of cesium from purex sludge supernatant at B Plant

International Nuclear Information System (INIS)

Carlstrom, R.F.

1977-01-01

Purex Sludge Supernatant (PSS) contains significant amounts of 137 Cs left after removal of strontium from fission product bearing Purex wastes. To remove cesium from PSS, an Ion Exchange Recovery system has been set up in Cells 17-21 at B Plant. The cesium that is recovered is stored within B Plant for eventual purification through the Cesium Purification process in Cell 38 and eventual encapsulation and storage in a powdered form at the Waste Encapsulation Storage Facility. Cesium depleted waste streams from the Ion Exchange processes are transferred to underground storage

Thrombin impairs human endometrial endothelial angiogenesis; implications for progestin-only contraceptive-induced abnormal uterine bleeding.

Science.gov (United States)

Shapiro, John P; Guzeloglu-Kayisli, Ozlem; Kayisli, Umit A; Semerci, Nihan; Huang, S Joseph; Arlier, Sefa; Larsen, Kellie; Fadda, Paolo; Schatz, Frederick; Lockwood, Charles J

2017-06-01

Progestin-only contraceptives induce abnormal uterine bleeding, accompanied by prothrombin leakage from dilated endometrial microvessels and increased thrombin generation by human endometrial stromal cell (HESC)-expressed tissue factor. Initial studies of the thrombin-treated HESC secretome identified elevated levels of cleaved chondroitin sulfate proteoglycan 4 (CSPG4), impairing pericyte-endothelial interactions. Thus, we investigated direct and CSPG4-mediated effects of thrombin in eliciting abnormal uterine bleeding by disrupting endometrial angiogenesis. Liquid chromatography/tandem mass spectrometry, enzyme-linked immunosorbent assay (ELISA) and quantitative real-time-polymerase chain reaction (PCR) evaluated conditioned medium supernatant and cell lysates from control versus thrombin-treated HESCs. Pre- and post-Depo medroxyprogesterone acetate (DMPA)-administered endometria were immunostained for CSPG4. Proliferation, apoptosis and tube formation were assessed in human endometrial endothelial cells (HEECs) incubated with recombinant human (rh)-CSPG4 or thrombin or both. Thrombin induced CSPG4 protein expression in cultured HESCs as detected by mass spectrometry and ELISA (pabnormal uterine bleeding in DMPA users. Mass spectrometry analysis identified several HESC-secreted proteins regulated by thrombin. Therapeutic agents blocking angiogenic effects of thrombin in HESCs can prevent or minimize progestin-only contraceptive-induced abnormal uterine bleeding. Copyright © 2017. Published by Elsevier Inc.

Isolation and Purification of an Early Pregnancy Factor–Like Molecule from Culture Supernatants Obtained from Lymphocytes of Pregnant Women

OpenAIRE

Aranha, Clara; Natraj, Usha; Iyer, K. S.; Shahani, Savitri

1998-01-01

Purpose:Our purpose was to determine whether lymphocytes synthesize proteins during pregnancy, to observe whether one of the proteins synthesized has early pregnancy factor (EPF)–like activity and to isolate and purify this molecule from culture supernatants obtained from stimulated lymphocytes of pregnant women.

Association of Atypical Enteropathogenic Escherichia coli with Diarrhea and Related Mortality in Kittens.

Science.gov (United States)

Watson, Victoria E; Jacob, Megan E; Flowers, James R; Strong, Sandra J; DebRoy, Chitrita; Gookin, Jody L

2017-09-01

Diarrhea is responsible for the death of approximately 900,000 children per year worldwide. In children, typical enteropathogenic Escherichia coli (EPEC) is a common cause of diarrhea and is associated with a higher hazard of death. Typical EPEC infection is rare in animals and poorly reproduced in experimental animal models. In contrast, atypical EPEC (aEPEC) infection is common in both children and animals, but its role in diarrhea is uncertain. Mortality in kittens is often attributed to diarrhea, and we previously identified enteroadherent EPEC in the intestines of deceased kittens. The purpose of this study was to determine the prevalence and type of EPEC in kittens and whether infection was associated with diarrhea, diarrhea-related mortality, gastrointestinal pathology, or other risk factors. Kittens with and without diarrhea were obtained from two shelter facilities and determined to shed atypical EPEC at a culture-based prevalence of 18%. In contrast, quantitative PCR detected the presence of the gene for intimin ( eae ) in feces from 42% of kittens. aEPEC was isolated from kittens with and without diarrhea. However, kittens with diarrhea harbored significantly larger quantities of aEPEC than kittens without diarrhea. Kittens with aEPEC had a significantly greater severity of small intestinal and colonic lesions and were significantly more likely to have required subcutaneous fluid administration. These findings identify aEPEC to be prevalent in kittens and a significant primary or contributing cause of intestinal inflammation, diarrhea, dehydration, and associated mortality in kittens. Copyright © 2017 American Society for Microbiology.

A Conditioned Medium of Umbilical Cord Mesenchymal Stem Cells Overexpressing Wnt7a Promotes Wound Repair and Regeneration of Hair Follicles in Mice

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Liang Dong

2017-01-01

Full Text Available Mesenchymal stem cells (MSCs can affect the microenvironment of a wound and thereby accelerate wound healing. Wnt proteins act as key mediators of skin development and participate in the formation of skin appendages such as hair. The mechanisms of action of MSCs and Wnt proteins on skin wounds are largely unknown. Here, we prepared a Wnt7a-containing conditioned medium (Wnt-CM from the supernatant of cultured human umbilical cord-MSCs (UC-MSCs overexpressing Wnt7a in order to examine the effects of this CM on cutaneous healing. Our results revealed that Wnt-CM can accelerate wound closure and induce regeneration of hair follicles. Meanwhile, Wnt-CM enhanced expression of extracellular matrix (ECM components and cell migration of fibroblasts but inhibited the migratory ability and expression of K6 and K16 in keratinocytes by enhancing expression of c-Myc. However, we found that the CM of fibroblasts treated with Wnt-CM (HFWnt-CM-CM can also promote wound repair and keratinocyte migration; but there was no increase in the number of hair follicles of regeneration. These data indicate that Wnt7a and UC-MSCs have synergistic effects: they can accelerate wound repair and induce hair regeneration via cellular communication in the wound microenvironment. Thus, this study opens up new avenues of research on the mechanisms underlying wound repair.

Carbon black nanoparticles induce type II epithelial cells to release chemotaxins for alveolar macrophages

Directory of Open Access Journals (Sweden)

Donaldson Ken

2005-12-01

Full Text Available Abstract Background Alveolar macrophages are a key cell in dealing with particles deposited in the lungs and in determining the subsequent response to that particle exposure. Nanoparticles are considered a potential threat to the lungs and the mechanism of pulmonary response to nanoparticles is currently under intense scrutiny. The type II alveolar epithelial cell has previously been shown to release chemoattractants which can recruit alveolar macrophages to sites of particle deposition. The aim of this study was to assess the responses of a type II epithelial cell line (L-2 to both fine and nanoparticle exposure in terms of secretion of chemotactic substances capable of inducing macrophage migration. Results Exposure of type II cells to carbon black nanoparticles resulted in significant release of macrophage chemoattractant compared to the negative control and to other dusts tested (fine carbon black and TiO2 and nanoparticle TiO2 as measured by macrophage migration towards type II cell conditioned medium. SDS-PAGE analysis of the conditioned medium from particle treated type II cells revealed that a higher number of protein bands were present in the conditioned medium obtained from type II cells treated with nanoparticle carbon black compared to other dusts tested. Size-fractionation of the chemotaxin-rich supernatant determined that the chemoattractants released from the epithelial cells were between 5 and 30 kDa in size. Conclusion The highly toxic nature and reactive surface chemistry of the carbon black nanoparticles has very likely induced the type II cell line to release pro-inflammatory mediators that can potentially induce migration of macrophages. This could aid in the rapid recruitment of inflammatory cells to sites of particle deposition and the subsequent removal of the particles by phagocytic cells such as macrophages and neutrophils. Future studies in this area could focus on the exact identity of the substance(s released by the

Study on bystander effect and associated mechanism mediated through culture medium

International Nuclear Information System (INIS)

Tu Xumin; Lei Suwen; Zhang Zhixing; Lv Huimin

2005-01-01

Objective: To study the bystander effect and associated mechanism mediated through the irradiated cell culture medium. Methods: Splenic natural killer (NK) cells were obtained from healthy male ICR strain mice. Culture medium irradiated with different doses of 60 Co γ-rays was used for culturing Yac-I lymphoma cells. The degree of injury of the latter by activated NK cells was observed. A part of the culture media were pretreated with 1% DMSO, a scavenger of reactive oxygen species (ROS), in order to investigate the possible mechanism of a radiation-induced bystander response. Results: Severer injury was induced in Yac-I cells cultured in the media pre-irradiated with different doses of γ-rays than that in Yac-I cells cultured in unirradiated medium, as shown by increased sensitivity to murine splenic NK cells (P<0.01). Culturing Yac-I cells in DMSO-pretreated medium considerably reduced the activation of NK cells, especially in 0.25 Gy and 0.5 Gy γ-irradiated media. Therefore, it can be expected that DMSO can partly suppress ROS-induced bystander effect. Conclusion: The irradiated culture medium of Yac-I cells can trigger bystander effect. ROS likely plays an important role in radiation-induced bystander effect that can be partly suppressed by pretreatment with DMSO. (authors)

Risk of contrast-medium-induced nephropathy in high-risk patients undergoing MDCT--a pooled analysis of two randomized trials

DEFF Research Database (Denmark)

Thomsen, Henrik S; Morcos, Sameh K

2009-01-01

of the LOCM were also observed in patients with GFR contrast agents in patients with moderate-to-severe renal insufficiency, the risk of significant CIN seems to be low. The IOCM iodixanol caused a higher rate of CIN than the LOCM......The incidence of contrast-medium-induced nephropathy (CIN) following intravenous (IV) CM administration of contrast media to renally impaired patients undergoing multidetector computed tomography (MDCT) is not well characterized. Our objective was to investigate the incidence of CIN in patients...... with glomerular filtration rate (GFR) contrast-enhanced MDCT examinations and to compare the rates of CIN following the IV administration of low-osmolar contrast media (LOCM, iopamidol and iomeprol) and an iso-osmolar contrast medium (IOCM, iodixanol). A total of 301 adult patients...

Medium-induced gluon radiation beyond the eikonal approximation

CERN Document Server

Apolinário, Liliana; Milhano, Guilherme; Salgado, Carlos A

2014-01-01

In this work we improve existing calculations of radiative energy loss by computing corrections that implement energy-momentum conservation, previously only implemented a posteriori, in a rigorous way. Using the path-integral formalism, we compute in-medium splittings allowing transverse motion of all particles in the emission process, thus relaxing the assumption that only the softest particle is permitted such movement. This work constitutes the extension of the computation carried out for x$\\rightarrow$1 in Phys. Lett. B718 (2012) 160-168, to all values of x, the momentum fraction of the energy of the parent parton carried by the emitted gluon. In order to accomplish a general description of the whole in-medium showering process, in this work we allow for arbitrary formation times for the emitted gluon. We provide general expressions and their realisation in the path integral formalism within the harmonic oscillator approximation.

Influence of Helicobacter pylori culture supernatant on the ecological balance of a dual-species oral biofilm

OpenAIRE

Wenling Zhang; Xiaohong Deng; Xuedong Zhou; Yuqing Hao; Yuqing Li

2018-01-01

Abstract Dental caries is a chronic progressive disease occurring in the tooth hard tissue due to multiple factors, in which bacteria are the initial cause. Both Streptococcus mutans and Streptococcus sanguinis are main members of oral biofilm. Helicobacter pylori may also be detected in dental plaque, playing an important role in the development of dental caries. Objective The aim of this study was to investigate the effect of H. pylori culture supernatant on S. mutans and S. sanguinis dual...

Mesenchymal stem cells and conditioned medium avert enteric neuropathy and colon dysfunction in guinea pig TNBS-induced colitis.

Science.gov (United States)

Robinson, Ainsley M; Sakkal, Samy; Park, Anthony; Jovanovska, Valentina; Payne, Natalie; Carbone, Simona E; Miller, Sarah; Bornstein, Joel C; Bernard, Claude; Boyd, Richard; Nurgali, Kulmira

2014-12-01

Damage to the enteric nervous system (ENS) associated with intestinal inflammation may underlie persistent alterations to gut functions, suggesting that enteric neurons are viable targets for novel therapies. Mesenchymal stem cells (MSCs) offer therapeutic benefits for attenuation of neurodegenerative diseases by homing to areas of inflammation and exhibiting neuroprotective, anti-inflammatory, and immunomodulatory properties. In culture, MSCs release soluble bioactive factors promoting neuronal survival and suppressing inflammation suggesting that MSC-conditioned medium (CM) provides essential factors to repair damaged tissues. We investigated whether MSC and CM treatments administered by enema attenuate 2,4,6-trinitrobenzene-sulfonic acid (TNBS)-induced enteric neuropathy and motility dysfunction in the guinea pig colon. Guinea pigs were randomly assigned to experimental groups and received a single application of TNBS (30 mg/kg) followed by 1 × 10(6) human bone marrow-derived MSCs, 300 μl CM, or 300 μl unconditioned medium 3 h later. After 7 days, the effect of these treatments on enteric neurons was assessed by histological, immunohistochemical, and motility analyses. MSC and CM treatments prevented inflammation-associated weight loss and gross morphological damage in the colon; decreased the quantity of immune infiltrate in the colonic wall (P ChAT, and nNOS immunoreactivity (P < 0.05); and alleviated inflammation-induced colonic dysmotility (contraction speed; P < 0.001, contractions/min; P < 0.05). These results provide strong evidence that both MSC and CM treatments can effectively prevent damage to the ENS and alleviate gut dysfunction caused by TNBS-induced colitis. Copyright © 2014 the American Physiological Society.

RNA-Seq analysis of isolate- and growth phase-specific differences in the global transcriptomes of enteropathogenic Escherichia coli prototype isolates

Science.gov (United States)

Hazen, Tracy H.; Daugherty, Sean C.; Shetty, Amol; Mahurkar, Anup A.; White, Owen; Kaper, James B.; Rasko, David A.

2015-01-01

Enteropathogenic Escherichia coli (EPEC) are a leading cause of diarrheal illness among infants in developing countries. E. coli isolates classified as typical EPEC are identified by the presence of the locus of enterocyte effacement (LEE) and the bundle-forming pilus (BFP), and absence of the Shiga-toxin genes, while the atypical EPEC also encode LEE but do not encode BFP or Shiga-toxin. Comparative genomic analyses have demonstrated that EPEC isolates belong to diverse evolutionary lineages and possess lineage- and isolate-specific genomic content. To investigate whether this genomic diversity results in significant differences in global gene expression, we used an RNA sequencing (RNA-Seq) approach to characterize the global transcriptomes of the prototype typical EPEC isolates E2348/69, B171, C581-05, and the prototype atypical EPEC isolate E110019. The global transcriptomes were characterized during laboratory growth in two different media and three different growth phases, as well as during adherence of the EPEC isolates to human cells using in vitro tissue culture assays. Comparison of the global transcriptomes during these conditions was used to identify isolate- and growth phase-specific differences in EPEC gene expression. These analyses resulted in the identification of genes that encode proteins involved in survival and metabolism that were coordinately expressed with virulence factors. These findings demonstrate there are isolate- and growth phase-specific differences in the global transcriptomes of EPEC prototype isolates, and highlight the utility of comparative transcriptomics for identifying additional factors that are directly or indirectly involved in EPEC pathogenesis. PMID:26124752

Comparison of DOT-ELISA and Standard-ELISA for Detection of the Vibrio cholerae Toxin in Culture Supernatants of Bacteria Isolated from Human and Environmental Samples.

Science.gov (United States)

Meza-Lucas, Antonio; Pérez-Villagómez, María-Fernanda; Martínez-López, José-Patricio; García-Rodea, Ricardo; Martínez-Castelán, María-Guadalupe; Escobar-Gutiérrez, Alejandro; de-la-Rosa-Arana, Jorge-Luis; Villanueva-Zamudio, Altagracia

2016-09-01

A comparison of DOT-ELISA and Standard-ELISA was made for detection of Vibrio cholerae toxin in culture supernatants of bacteria isolated from human and environmental samples. A total of 293 supernatants were tested in a double blind assay. A correlation of 100 % was obtained between both techniques. The cholera toxin was found in 20 Inaba and 3 Ogawa strains. Positive samples were from seafood (17 samples), potable water (1 sample) and sewage (5 samples). The DOT-ELISA was useful as the standard-ELISA to confirm the presence of cholera toxin in the environmental samples.

Macrophages are related to goblet cell hyperplasia and induce MUC5B but not MUC5AC in human bronchus epithelial cells.

Science.gov (United States)

Silva, Manuel A; Bercik, Premysl

2012-06-01

Airway goblet cell hyperplasia (GCH)--detectable by mucin staining--and abnormal macrophage infiltrate are pathological features present in many chronic respiratory disorders. However, it is unknown if both factors are associated. Using in-vivo and in-vitro models, we investigated whether macrophages are related with GCH and changes in mucin immunophenotypes. Lung sections from Sprague-Dawley rats treated for 48 h with one intra-tracheal dose of PBS or LPS (n=4-6 per group) were immunophenotyped for rat-goblet cells, immune, and proliferation markers. Human monocyte-derived macrophages (MDM) were pre-treated with or without LPS, immunophenotyped, and their supernatant, as well as cytokines at levels equivalent to supernatant were used to challenge primary culture of normal human bronchus epithelial cells (HBEC) in air-liquid interface, followed by MUC5B and MUC5AC mucin immunostaining. An association between increased bronchiolar goblet cells and terminal-bronchiolar proliferative epithelial cells confirmed the presence of GCH in our LPS rat model, which was related with augmented bronchiolar CD68 macrophage infiltration. The in-vitro experiments have shown that MUC5AC phenotype was inhibited when HBEC were challenged with supernatant from MDM pre-treated with or without LPS. In contrast, TNF-α and interleukin-1β at levels equivalent to supernatant from LPS-treated MDM increased MUC5AC. MUC5B was induced by LPS, supernatant from LPS-treated MDM, a mix of cytokines including TNF-α and TNF-α alone at levels present in supernatant from LPS-treated MDM. We demonstrated that macrophages are related with bronchiolar GCH, and that they induced MUC5B and inhibited MUC5AC in HBEC, suggesting a role for them in the pathogenesis of airway MUC5B-related GCH.

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

Science.gov (United States)

Yao, Yongchang; Zhai, Zhichen; Wang, Yingjun

2014-01-01

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

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

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Yongchang Yao

2014-01-01

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

Peroxynitrite decomposition catalyst prevents apoptotic cell death in a human astrocytoma cell line incubated with supernatants of HIV-infected macrophages

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Perno Carlo

2002-09-01

Full Text Available Abstract Background Oxidative stress has shown to contribute in the mechanisms underlying apoptotic cell death occuring in AIDS-dementia complex. Here we investigated the role of peroxynitrite in apoptosis occurring in astroglial cells incubated with supernatants of HIV-infected human primary macrophages (M/M. Results Flow cytometric analysis (FACS of human cultured astrocytes shortly incubated with HIV-1-infected M/M supernatants showed apoptotic cell death, an effect accompanied by pronounced staining for nitrotyrosine (footprint of peroxynitrite and by abnormal formation of malondialdehyde (MDA. Pretreatment of astrocytes with the peroxynitrite decomposition catalyst FeTMPS antagonized HIV-related astrocytic apoptosis, MDA formation and nitrotyrosine staining. Conclusions Taken together, our results suggest that inibition of peroxynitrite leads to protection against peroxidative stress accompanying HIV-related apoptosis of astrocytes. Overall results support the role of peroxynitrite in HIV-related programmed death of astrocytes and suggest the use of peroxynitrite decomposition catalyst to counteract HIV-1-related neurological disorders.

Quantitation, in vitro propagation, and characterization of preleukemic cells induced by radiation leukemia virus

International Nuclear Information System (INIS)

Yefenof, E.; Epszteyn, S.; Kotler, M.

1991-01-01

Intrathymic (i.t.) inoculation of radiation leukemia virus into C57BL/6 mice induces a population of preleukemic (PL) cells that can progress into mature thymic lymphomas upon transfer into syngeneic recipients. A minimum of 10(3) PL thymic cells are required to induce lymphomas in the recipient. Most of the individual lymphomas developed in mice which were inoculated with cells of a single PL thymus, derived from different T-cell precursors. PL thymic cells could be grown in vitro on a feeder layer consisting of splenic stromal cells. Growth medium was supplemented with supernatant harvested from an established radiation leukemia virus-induced lymphoma cell line (SR4). The in vitro-grown PL cells were characterized as Thy-1+, CD4+, CD8- T-cells, most of which expressed radiation leukemia virus antigens. Cultured PL cells were found to be nontumorigenic, based on their inability to form s.c. tumors. However, these cells could develop into thymic lymphomas if inoculated i.t. into syngeneic recipients. A culture of PL cells, maintained for 2 mo, showed clonal T-cell receptor arrangement. Lymphomas which developed in several recipient mice upon injection with these PL cells were found to possess the same T-cell receptor arrangement. These results indicate that PL cells can be adapted for in vitro growth while maintaining their preleukemic character

In vitro and in vivo antitumor effects of 50 to 100-KDa components from B16 melanoma culture supernatant.

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Qin, Ying-Song; Zhang, X U; Zhang, Xiang-Yu

2015-07-01

The development of immunological therapies for melanoma has been of considerable concern in recent years. Whole tumor cell lysates have been used to develop antitumor vaccines, but the effective components of the lysates have not been identified. In the present study, protein elements were purified from the B16 supernatant to analyze the in vitro chemotaxis towards mouse spleen lymphocytes using a Boyden chamber. Prior to establishing a B16 melanoma model, C57BL/6 mice were vaccinated with these proteins, and melanoma growth, tumor appearance time and behavioral changes were observed. Next, the cytotoxicity and subsets of the tumor infiltrating lymphocytes, and the histological characteristics of the melanoma were analyzed. The isolated purified fragments of B16 melanoma culture supernatant had strong antitumor effects. The possible antitumor mechanism was delineated, and was identified to possibly be through the activation of cluster of differentiation 8-positive T cells and the promotion of B16 cell differentiation. These methods will provide a novel insight into understanding antitumor immunological mechanisms and provide a potential avenue for immunotherapy.

Performance of hemicellulolytic enzymes in culture supernatants from a wide range of fungi on insoluble wheat straw and corn fiber fractions

NARCIS (Netherlands)

Gool, van M.P.; Toth, K.; Schols, H.A.; Szakacs, G.; Gruppen, H.

2012-01-01

Filamentous fungi are a good source of hemicellulolytic enzymes for biomass degradation. Enzyme preparations were obtained as culture supernatants from 78 fungal isolates grown on wheat straw as carbon source. These enzyme preparations were utilized in the hydrolysis of insoluble wheat straw and

Protective Effects of Decay-Accelerating Factor on Blast-Induced Neurotrauma in Rats

Science.gov (United States)

2013-08-16

gelatin -Veronal buffer (pH 7.3). After centrifugation, absorbance of the supernatant was determined at 405 nm, and the serum concentration inducing 50...Houston, TX 78234, USA. 2Department of Trauma and Resuscitation Medicine, Naval Medical Research Center, 503 Robert Grant Avenue, Silver Spring, MD

Structure of nitrogen-converting communities induced by hydraulic retention time and COD/N ratio in constantly aerated granular sludge reactors treating digester supernatant.

Science.gov (United States)

Cydzik-Kwiatkowska, Agnieszka; Rusanowska, Paulina; Zielińska, Magdalena; Bernat, Katarzyna; Wojnowska-Baryła, Irena

2014-02-01

This study investigated how hydraulic retention time (HRT) and COD/N ratio affect nitrogen-converting consortia in constantly aerated granules treating high-ammonium digester supernatant. Three HRTs (10, 13, 19 h) were tested at COD/N ratios of 4.5 and 2.3. Denaturing gradient gel electrophoresis and relative real-time PCR were used to characterize the microbial communities. When changes in HRT and COD/N increased nitrogen loading, the ratio of the relative abundance of aerobic to anaerobic ammonium-oxidizers decreased. The COD/N ratio determined the species composition of the denitrifiers; however, Thiobacillus denitrificans, Pseudomonas denitrificans and Azoarcus sp. showed a high tolerance to the environmental conditions and occurred in the granules from all reactors. Denitrifier genera that support granule formation were identified, such as Pseudomonas, Shinella, and Flavobacterium. In aerated granules, nirK-possessing bacteria were more diverse than nirS-possessing bacteria. At a low COD/N ratio, N2O-reducer diversity increased because of the presence of bacteria known as aerobic denitrifiers. Copyright © 2013 Elsevier Ltd. All rights reserved.

Attaching and effacing Escherichia coli isolates from Danish children: clinical significance and microbiological characteristics

DEFF Research Database (Denmark)

Jensen, C; Ethelberg, S; Olesen, B

2007-01-01

This study describes the prevalence, clinical manifestations and microbiological characteristics of attaching and effacing Escherichia coli isolates, i.e., enteropathogenic E. coli (EPEC) belonging to the classical EPEC serotypes, non-EPEC attaching and effacing E. coli (A/EEC) and verocytotoxin...

Effects of mesenchymal stem cells conditioned medium on behavioral aspects of inflammatory arthritic pain induced by CFA adjuvant

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Vida Nazemian

2016-07-01

Full Text Available Background: Rheumatoid arthritis is a type of inflammatory pain and is an autoimmune and chronic inflammatory disease which can lead to hyperalgesia, edema and decreased motor activity in affected area. Mesenchymal stem cells conditioned medium (MSC-CM has anti-inflammatory mediators which can regulate the immune responses, alleviate inflammatory symptoms and has a paracrine effects too. The aim of this study was to evaluate the effects of mesenchymal stem cells conditioned medium on behavioral aspects of inflammatory arthritic pain which induced by CFA adjuvant.Materials and Methods: Complete Freund’s adjuvant (CFA-induced arthritis (AA was caused by single subcutaneous injection of CFA into the rats hind paw on day zero. MSC-CM was administered daily and intraperitoneal during the 21 days of the study after CFA injection. Hyperalgesia and edema were assessed on days 0, 7, 14 and 21 of the study respectively with radian heat and plethysmometer instrument.Results: The results of this study indicated the significant roles of MSC-CM in betterment of inflammatory symptoms such as hyperalgesia and edema during different stages of inflammation caused by CFA. The continuing injection of MSC-CM could reduce the inflammatory symptoms.Conclusion: Long term treatment by MSC-CM can alleviate hyperalgesia and edema and decrease those to the level of the time before induction of inflammation.   

Putative new heat-stable cytotoxic and enterotoxic factors in culture supernatant of Escherichia coli isolated from drinking water

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DA Ribeiro

2011-01-01

Full Text Available Enteric infections caused by the ingestion of contaminated water, especially by Escherichia coli, are important to define the virulence properties of these bacteria. Due to frequent infantile diarrhea in the city of Ouro Preto, Minas Gerais state, Brazil, the phenotypic and genotypic diarrheagenic properties of E. coli isolated from drinking water were studied. The culture supernatants of 39 (40% among a total of 97 E. coli isolates from drinking water were positive by suckling mouse assay and induced cytotoxic effects on Vero cells. The enterotoxic and cytotoxic activities were present in the fraction with less than 10 kDa and were not lost when heated up to 60°C and 100°C for 30 minutes. PCR assays showed that among these 39 Vero cytotoxigenic E. coli, four (10.2% were positive for ST II (estB and two (5% positive for αHly (hlyA. Gene amplification of SLT (stx 1, stx 2, ST I (estA, LT (eltI, eltII, EAST1 (astA, EHly (enhly and plasmid-encoded enterotoxin (pet were not observed. This heat-stable cytotoxic enterotoxin of E. coli is probably a new putative diarrheagenic virulence factor, as a toxin presenting these characteristics has not yet been described.

Risk of application of contrast medium in computed tomography

Energy Technology Data Exchange (ETDEWEB)

Wende, S.; Speck, U.

1981-06-01

The paper deals with the risk associated with the application of contrast medium in CT. The risk is to be seen in intolerance reactions, such as allergic or circulatory reactions and neurotoxic side-effects. In this paper the problems of renal failure caused by the injection of contrast medium are given special attention. Furthermore an iodine-induced hyperthyreosis might result. Especially the possible disturbance of the kidney function means that contrast medium should not be applied arbitrarily and that the examination should be done only by experienced staff. Furthermore the indication for the application of contrast medium in CT should be strictly qualified.

Methodology for monitoring gold nanoparticles and dissolved gold species in culture medium and cells used for nanotoxicity tests by liquid chromatography hyphenated to inductively coupled plasma-mass spectrometry.

Science.gov (United States)

López-Sanz, Sara; Fariñas, Nuria Rodríguez; Vargas, Rosario Serrano; Martín-Doimeadios, Rosa Del Carmen Rodríguez; Ríos, Ángel

2017-03-01

An analytical methodology based on coupling reversed-phase liquid chromatography (HPLC) to an inductively coupled plasma mass spectrometry (ICP-MS) has been developed for the characterization and identification of gold nanoparticles (AuNPs) and gold dissolved species (Au 3+ ) in culture medium (Dulbecco's Modified Eagle Medium, DMEM) and HeLa cells (a human cervical adenocarcinoma cell line) used in nanotoxicity tests. The influence of the culture medium was also studied and the method applied for nanotoxicity tests. It was also observed that AuNPs can undergo an oxidation process in the supernatants and only a small amount of AuNPs and dissolved Au 3+ was associated with cells. To evaluate the biological impact of AuNPs, a classical viability assay onto HeLa cells was performed using cellular media DMEM in the presence of increasing dosage of 10nm AuNPs. The results showed that 10nm AuNPs exhibit a slight toxic effect. Copyright © 2016 Elsevier B.V. All rights reserved.

Cultivation of Scenedesmus dimorphus using anaerobic digestate as a nutrient medium.

Science.gov (United States)

Abu Hajar, Husam A; Riefler, R Guy; Stuart, Ben J

2017-08-01

In this study, the microalga Scenedesmus dimorphus was cultivated phototrophically using unsterilized anaerobic digestate as a nutrient medium. A bench-scale experiment was conducted by inoculating the microalga S. dimorphus with 0.05-10% dilutions of the anaerobic digestate supernatant. It was found that 1.25-2.5% dilutions, which is equivalent to 50-100 mg N/L total nitrogen concentrations and 6-12 mg P/L total phosphorus concentrations, provided sufficient nutrients to maximize the growth rate along with achieving high concentrations of algal biomass. The microalgae cultivation was scaled up to 100 L open raceway ponds, where the effect of paddlewheel mixing on the growth was investigated. It was concluded that 0.3 m/s water surface velocity yielded the highest specific growth rate and biomass concentration compared to 0.1 and 0.2 m/s. The microalga S. dimorphus was then cultivated in the raceway ponds using 2.5% diluted anaerobic digestate at 317 and 454 μmol/(m 2  × s) average incident light intensities and 1.25% diluted anaerobic digestate at 234 and 384 μmol/(m 2  × s) average incident light intensities. The maximum biomass concentration was 446 mg/L which was achieved in the 2.5% dilution and 454 μmol/(m 2  × s) light intensity culture. Moreover, nitrogen, phosphorus, and COD removal efficiencies from the nutrient media were 65-72, 63-100, and 78-82%, respectively, whereas ammonia was completely removed from all cultures. For a successful and effective cultivation in open raceway ponds, light intensity has to be increased considerably to overcome the attenuation caused by the algal biomass as well as the suspended solids from the digestate supernatant.

Aspirin Inhibits Platelet-Derived Sphingosine-1-Phosphate Induced Endothelial Cell Migration.

Science.gov (United States)

Polzin, Amin; Knoop, Betül; Böhm, Andreas; Dannenberg, Lisa; Zurek, Mark; Zeus, Tobias; Kelm, Malte; Levkau, Bodo; Rauch, Bernhard H

2018-01-01

Aspirin plays a crucial role in the prevention of cardiovascular diseases. We previously described that aspirin has effects beyond inhibition of platelet aggregation, as it inhibited thrombin-mediated release of sphingosine-1-phosphate (S1P) from human platelets. S1P is a bioactive lipid with important functions on inflammation and apoptosis. In endothelial cells (EC), S1P is a key regulator of cell migration. In this study, we aimed to analyze the effects of aspirin on platelet-induced EC migration. Human umbilical EC migration was measured by Boyden chamber assay. EC migration was induced by platelet supernatants of thrombin receptor-activating peptide-1 (AP1) stimulated platelets. To investigate the S1P receptor subtype that promotes EC migration, specific inhibitors of S1P receptor subtypes were applied. S1P induced EC migration in a concentration-dependent manner. EC migration induced by AP1-stimulated platelet supernatants was reduced by aspirin. S1P1 receptor inhibition almost completely abolished EC migration induced by activated platelets. The inhibition of S1P2 or S1P3 receptor had no effect. Aspirin inhibits EC migration induced by activated platelets that is in part due to S1P and mediated by the endothelial S1P1 receptor. The clinical significance of this novel mechanism of aspirin action has to be investigated in future studies. © 2017 S. Karger AG, Basel.

Dynamics of E.coli virulence factors in dairy cow herds

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Background. Dairy farms are known reservoirs of entero-pathogenic E. coli (EPEC). EPEC, or the virulence factors associated with pathogenicity, have been detected in manure, milk, and the farm environment. However, it is unclear which farm compartments are reservoirs contributing to EPEC persistence...

In Vitro Dialysis of Cytokine-Rich Plasma With High and Medium Cut-Off Membranes Reduces Its Procalcific Activity.

Science.gov (United States)

Willy, Kevin; Hulko, Michael; Storr, Markus; Speidel, Rose; Gauss, Julia; Schindler, Ralf; Zickler, Daniel

2017-09-01

Recently developed high-flux (HF) dialysis membranes with extended permeability provide better clearance of middle-sized molecules such as interleukins (ILs). Whether this modulation of inflammation influences the procalcific effects of septic plasma on vascular smooth muscle cells (VSMCs) is not known. To assess the effects of high cut-off (HCO) and medium cut-off (MCO) membranes on microinflammation and in vitro vascular calcification we developed a miniature dialysis model. Plasma samples from lipopolysaccharide-spiked blood were dialyzed with HF, HCO, and MCO membranes in an in vitro miniature dialysis model. Afterwards, IL-6 concentrations were determined in dialysate and plasma. Calcifying VSMCs were incubated with dialyzed plasma samples and vascular calcification was assessed. Osteopontin (OPN) and matrix Gla protein (MGP) were measured in VSMC supernatants. IL-6 plasma concentrations were markedly lower with HCO and MCO dialysis. VSMC calcification was significantly lower after incubation with MCO- and HCO-serum compared to HF plasma. MGP and OPN levels in supernatants were significantly lower in the MCO but not in the HCO group compared to HF. In vitro dialysis of cytokine-enriched plasma samples with MCO and HCO membranes reduces IL-6 levels. The induction of vascular calcification by cytokine-enriched plasma is reduced after HCO and MCO dialysis. © 2017 International Center for Artificial Organs and Transplantation and Wiley Periodicals, Inc.

Evaluation of Synergistic Interactions Between Cell-Free Supernatant of Lactobacillus Strains and Amikacin and Genetamicin Against Pseudomonas aeruginosa

OpenAIRE

Aminnezhad, Sargol; Kermanshahi, Rouha Kasra; Ranjbar, Reza

2015-01-01

Background: The indiscriminate use of antibiotics in the treatment of infectious diseases can increase the development of antibiotic resistance. Therefore, there is a big demand for new sources of antimicrobial agents and alternative treatments for reduction of antibiotic dosage required to decrease the associated side effects. Objectives: In this study, the synergistic action of aminoglycoside antibiotics and cell-free supernatant (CFS) of probiotic (Lactobacillus rahmnosus and L. casei) aga...

Transformation of carbon tetrachloride and chloroform by trichloroethene respiring anaerobic mixed cultures and supernatant.

Science.gov (United States)

Vickstrom, Kyle E; Azizian, Mohammad F; Semprini, Lewis

2017-09-01

Carbon tetrachloride (CT) and chloroform (CF) were transformed in batch reactor experiments conducted with anaerobic dechlorinating cultures and supernatant (ADC + S) harvested from continuous flow reactors. The Evanite (EV) and Victoria/Stanford (VS) cultures, capable of respiring trichloroethene (TCE), 1,2-cis-dichloroethene (cDCE), and vinyl chloride (VC) to ethene (ETH), were grown in continuous flow reactors receiving an influent feed of saturated TCE (10 mM; 60 mEq) and formate (45 mM; 90 mEq) but no CT or CF. Cells and supernatant were harvested from the chemostats and inoculated into batch reactors at the onset of each experiment. CT transformation was complete following first order kinetics with CF, DCM and CS 2 as the measurable transformation products, representing 20-40% of the original mass of CT, with CO 2 likely the unknown transformation product. CF was transformed to DCM and likely CO 2 at an order of magnitude rate lower than CT, while DCM was not further transformed. An analytical first order model including multiple key reactions effectively simulated CT transformation, product formation and transformation, and provided reasonable estimates of transformation rate coefficients. Biotic and abiotic treatments indicated that CT was mainly transformed via abiotic processes. However, the presence of live cells was associated with the transformation of CF to DCM. In biotic tests both TCE and CT were simultaneously transformed, with TCE transformed to ETH and approximately 15-53% less CF formed via CT transformation. A 14-day exposure to CF (CF max  = 1.4 μM) reduced all rates of chlorinated ethene respiration by a factor of 10 or greater. Copyright © 2017 Elsevier Ltd. All rights reserved.

Host markers in Quantiferon supernatants differentiate active TB from latent TB infection: preliminary report

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Walzl Gerhard

2009-05-01

Full Text Available Abstract Background Interferon gamma release assays, including the QuantiFERON® TB Gold In Tube (QFT have been shown to be accurate in diagnosing Mycobacterium tuberculosis infection. These assays however, do not discriminate between latent TB infection (LTBI and active TB disease. Methods We recruited twenty-three pulmonary TB patients and 34 household contacts from Cape Town, South Africa and performed the QFT test. To investigate the ability of new host markers to differentiate between LTBI and active TB, levels of 29 biomarkers in QFT supernatants were evaluated using a Luminex multiplex cytokine assay. Results Eight out of 29 biomarkers distinguished active TB from LTBI in a pilot study. Baseline levels of epidermal growth factor (EGF soluble CD40 ligand (sCD40L, antigen stimulated levels of EGF, and the background corrected antigen stimulated levels of EGF and macrophage inflammatory protein (MIP-1β were the most informative single markers for differentiation between TB disease and LTBI, with AUCs of 0.88, 0.84, 0.87, 0.90 and 0.79 respectively. The combination of EGF and MIP-1β predicted 96% of active TB cases and 92% of LTBIs. Combinations between EGF, sCD40L, VEGF, TGF-α and IL-1α also showed potential to differentiate between TB infection states. EGF, VEGF, TGF-α and sCD40L levels were higher in TB patients. Conclusion These preliminary data suggest that active TB may be accurately differentiated from LTBI utilizing adaptations of the commercial QFT test that includes measurement of EGF, sCD40L, MIP-1β, VEGF, TGF-α or IL-1α in supernatants from QFT assays. This approach holds promise for development as a rapid diagnostic test for active TB.

Diabetes-Induced Oxidative Stress in Endothelial Progenitor Cells May Be Sustained by a Positive Feedback Loop Involving High Mobility Group Box-1

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Han Wu

2016-01-01

Full Text Available Oxidative stress is considered to be a critical factor in diabetes-induced endothelial progenitor cell (EPC dysfunction, although the underlying mechanisms are not fully understood. In this study, we investigated the role of high mobility group box-1 (HMGB-1 in diabetes-induced oxidative stress. HMGB-1 was upregulated in both serum and bone marrow-derived monocytes from diabetic mice compared with control mice. In vitro, advanced glycation end productions (AGEs induced, expression of HMGB-1 in EPCs and in cell culture supernatants in a dose-dependent manner. However, inhibition of oxidative stress with N-acetylcysteine (NAC partially inhibited the induction of HMGB-1 induced by AGEs. Furthermore, p66shc expression in EPCs induced by AGEs was abrogated by incubation with glycyrrhizin (Gly, while increased superoxide dismutase (SOD activity in cell culture supernatants was observed in the Gly treated group. Thus, HMGB-1 may play an important role in diabetes-induced oxidative stress in EPCs via a positive feedback loop involving the AGE/reactive oxygen species/HMGB-1 pathway.

Comparison of renal toxicity after injection of CT contrast medium and MR contrast medium: change of renal function in acute renal failure rat models

International Nuclear Information System (INIS)

Han, Young min; Lee, Young Hwan; Kim, Sang Won; Jin, Kong Young; Kim, Won; Chung, Gyung Ho

2002-01-01

To determine renal toxicity through changes in renal function after the injection of CT and MRI contrast media into rats in which acute renal failure (ARF) was induced. To cause acute renal failure, the abdominal cavity of 110 male rats each weighing 250-300 gm was opened via a midline incision under anesthesia. Microvascular clamps were placed on both renal arteries and veins to completely block renal blood flow for 45 minutes, and were then removed, allowing blood flow to return to the kidneys. ARF, defined as a two-fold difference in the creatinine level before ARF and 48 hours after, was successfully induced in 60 of the rats. These were divided into two groups: one was injected with CT contrast medium and the other with MRI contrast medium. Each CT and MRI group was divided into a low dose (0.5 cc/kg, 0.2 ml/kg), standard dose (2 cc/kg, 0.8 ml/kg), and high dose (8 cc/kg, 3.2 ml/kg) sub-group; thus, there was a total of six groups with ten rats in each. Blood samples were obtained before ARF, 48 hours after, and 48 hours after contrast injection, and CT scanning and MRI were performed after blood sampling at 48 hours. In each group, creatinine levels 48 hours after contrast injection were compared by means of the ANOVA test. There were no significant differences in creatinine levels between the CT and MRI contrast medium groups (p=0.116), nor between the animals to which different doses of CT and MRI contrast medium, were administered. After both standard and high doses, CT and MRI provided good images. In rats in which acute renal failure was induced, renal function did not change according to whether CT or MRI contrast medium was injected. Thus, the two media induce similar levels of toxicity

Epithelial cells detect functional type III secretion system of enteropathogenic Escherichia coli through a novel NF-κB signaling pathway.

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Yael Litvak

2017-07-01

Full Text Available Enteropathogenic Escherichia coli (EPEC, a common cause of infant diarrhea, is associated with high risk of mortality in developing countries. The primary niche of infecting EPEC is the apical surface of intestinal epithelial cells. EPEC employs a type three secretion system (TTSS to inject the host cells with dozens of effector proteins, which facilitate attachment to these cells and successful colonization. Here we show that EPEC elicit strong NF-κB activation in infected host cells. Furthermore, the data indicate that active, pore-forming TTSS per se is necessary and sufficient for this NF-κB activation, regardless of any specific effector or protein translocation. Importantly, upon infection with wild type EPEC this NF-κB activation is antagonized by anti-NF-κB effectors, including NleB, NleC and NleE. Accordingly, this NF-κB activation is evident only in cells infected with EPEC mutants deleted of nleB, nleC, and nleE. The TTSS-dependent NF-κB activation involves a unique pathway, which is independent of TLRs and Nod1/2 and converges with other pathways at the level of TAK1 activation. Taken together, our results imply that epithelial cells have the capacity to sense the EPEC TTSS and activate NF-κB in response. Notably, EPEC antagonizes this capacity by delivering anti-NF-κB effectors into the infected cells.

Allergen-stimulated T lymphocytes from allergic patients induce vascular cell adhesion molecule-1 (VCAM-1) expression and IL-6 production by endothelial cells.

Science.gov (United States)

Delneste, Y; Jeannin, P; Gosset, P; Lassalle, P; Cardot, E; Tillie-Leblond, I; Joseph, M; Pestel, J; Tonnel, A B

1995-01-01

Adhesion of inflammatory cells to endothelium is a critical step for their transvascular migration to inflammatory sites. To evaluate the relationship between T lymphocytes (TL) and vascular endothelium, supernatants from allergen-stimulated TL obtained from patients sensitive to Dermatophagoides pteronyssinus (Dpt) versus healthy subjects were added to endothelial cell (EC) cultures. TL were stimulated by autologous-activated antigen-presenting cells (APC) previously fixed in paraformaldehyde to prevent monokine secretion. Two parameters were measured: the expression of adhesion molecule and the production of IL-6. Related allergen-stimulated TL supernatants from allergic patients induced an increase of VCAM-1 and intercellular adhesion molecule-1 (ICAM-1) expression when supernatants of the control groups (TL exposed to an unrelated allergen or not stimulated or TL obtained from healthy subjects) did not. E-selectin expression was not modulated whatever the supernatant added to EC culture. IL-6 production by EC was significantly enhanced after activation with related allergen-stimulated TL supernatants from allergics compared with control supernatants. Induction of VCAM-1 expression was inhibited by adding neutralizing antibodies against IL-4, whereas IL-6 production and ICAM-1 expression were inhibited by anti-interferon-gamma (IFN-gamma) antibodies. Enhanced production of IL-4 and IFN-gamma was detected in related allergen-stimulated TL supernatants from allergic subjects compared with the different supernatants. These data suggest that allergen-specific TL present in the peripheral blood of allergic patients are of Th1 and Th2 subtypes. Their stimulation in allergic patients may lead to the activation of endothelial cells and thereby participate in leucocyte recruitment towards the inflammatory site. PMID:7542574

(EPEC) from pig

African Journals Online (AJOL)

SAM

2014-03-26

Mar 26, 2014 ... 16 belonged to O60 (94.1%) and 1(5.88%) was untypeable. Virulence genes ..... Figure 4. a) Confluent monolayer of healthy vero cells; b) CPE in verocells after 12 h of incubation with filtrate extracts .... about 5 to 10 pigs near their residence and share the ... of Stx to vero cells remains the 'gold standard' for.

Effect of Mg2+ and Fe2+ Concentrations in Culture Medium on CGF Formation from Microalgae Chlorella Pyrenoidosa Ink and Analysis of Amino Acids by Liquid Chromatography-Mass Spectrometry

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Kusmiati Kusmiati

2015-06-01

Full Text Available Chlorella pyrenoidosa (C. pyrenoidosa contains Chlorella Growth Factor (CGF, which consists of proteins and polysaccharides. CGF is located inside the nucleus of cells and is beneficial to humans as a food supplement, an immunity booster, and an antioxidant. CGF formation of C. pyrenoidosa is influenced by medium composition. C. pyrenoidosa INK was cultured in a modified basal medium (MBM with various concentrations of Mg2+ (0.5, 1.0, and 1.5 g/L and Fe2+ (3.5×10-4 and 5.0×10-4 g/L. The experiments were performed and analyzed under a completely randomized design using a 2-L bottle with three replications. The results showed that MBM with 1.0 g/L of Mg2+ and 3.5×10-4 g/L of Fe2+ yielded the optimal growth curve for C. pyrenoidosa. Analysis of protein content was carried out using the Lowry method with a spectrophotometer at λ=750 nm, and the obtained results were 0.0974 mg/mL (extract and 6.4097 mg/ml (supernatant. Furthermore, analysis of glucose content was carried out using the phenol sulfate method (λ = 490 nm, and the obtained results were 49.331 ppm (extract and 1566.911 ppm (supernatant. Analysis of amino acids in CGF using liquid chromatography-mass spectrometry (LC-MS indicated the presence of tyrosine, proline, glutamate, alanine, valine, tryptopan, phenylalanine, methionine, and leucine-isoleucine.

Risk of application of contrast medium in computed tomography

International Nuclear Information System (INIS)

Wende, S.; Speck, U.; Schering A.G., Berlin

1981-01-01

The paper deals with the risk associated with the application of contrast medium in CT. The risk is to be seen in intolerance reactions, such as allergic or circulatory reactions and neurotoxic side-effects. In this paper the problems of renal failure caused by the injection of contrast medium are given special attention. Furthermore an iodine-induced hyperthyreosis might result. Especially the possible disturbance of the kidney function means that contrast medium should not be applied arbitrarily and that the examination should be done only by experienced staff. Furthermore the indication for the application of contrast medium in CT should be strictly qualified. (orig.) [de

In-medium effects around the Fermi energy

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Lopez O.

2015-01-01

Full Text Available We study nuclear stopping in central collisions for heavy-ion induced reactions in the Fermi energy domain (15-100 AMeV. Using the large dataset provided by the 4π array INDRA, we determine that stopping can be directly related to the transport properties in the nuclear medium. By looking specifically at protons, we present a comprehensive body of experimental results concerning the mean free path, the nucleon-nucleon cross-section and in-medium effects in nuclear matter.

Isotope inequilibrium of glucose metabolites in intact cells and particlefree supernatants of Ehrlich ascites tumor

International Nuclear Information System (INIS)

Daehnfeldt, J.L.; Winge, P.

1975-01-01

With an enzyme degradative technique, isotope inequilibrium of glucose metabolites was demonstrated in intact cells and particle-free supernatants of Ehrlich ascites tumor using I- 14 C-glucose as tracer. Inequilibrium was found between glucose and glucose-6-phosphate, glucose and fructose-6-phosphate, glucose and 6-phosphogluconate, while glucose-6-phosphate and fructose-6-phosphate were found to be in near equilibrium within the incubation time investigated. Glucose and lactate were found to be in near equilibrium after 8 min in intact cells. Calculations based on the equilibrium levels found, showed that these inequilibria could not be explained by the effects of the pentose cycle. (U.S.)

Use of Gifu Anaerobic Medium for culturing 32 dominant species of human gut microbes and its evaluation based on short-chain fatty acids fermentation profiles.

Science.gov (United States)

Gotoh, Aina; Nara, Misaki; Sugiyama, Yuta; Sakanaka, Mikiyasu; Yachi, Hiroyuki; Kitakata, Aya; Nakagawa, Akira; Minami, Hiromichi; Okuda, Shujiro; Katoh, Toshihiko; Katayama, Takane; Kurihara, Shin

2017-10-01

Recently, a "human gut microbial gene catalogue," which ranks the dominance of microbe genus/species in human fecal samples, was published. Most of the bacteria ranked in the catalog are currently publicly available; however, the growth media recommended by the distributors vary among species, hampering physiological comparisons among the bacteria. To address this problem, we evaluated Gifu anaerobic medium (GAM) as a standard medium. Forty-four publicly available species of the top 56 species listed in the "human gut microbial gene catalogue" were cultured in GAM, and out of these, 32 (72%) were successfully cultured. Short-chain fatty acids from the bacterial culture supernatants were then quantified, and bacterial metabolic pathways were predicted based on in silico genomic sequence analysis. Our system provides a useful platform for assessing growth properties and analyzing metabolites of dominant human gut bacteria grown in GAM and supplemented with compounds of interest.

Fibroblast and T cells conditioned media induce maturation dendritic cell and promote T helper immune response

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Masoumeh Asadi

2012-06-01

Full Text Available Dendritic cells (DCs induce pathogen-specific T cell responses. We comprehensively studied the effects of addition of maturation stimulus, fibroblasts (fibroblast conditioned medium, PHA activated T cells (T cell conditioned medium, and mixture of fibroblast & PHA activated T cells (FCM-TCCM conditioned media on maturation of DCs. Monocytes were cultured with GM-CSF and IL-4 for five days. Maturation factors included MCM and TNF-α as control group. FCM and TCCM, or FCM-TCCM supernatant were considered as the treatment group. Tumor antigens were added at day five. Matured DCs were harvested at day seven. Phenotypic and functional analyses were carried out using anti (CD14, CD80, CD86, CD83 and HLA-DR monoclonal antibodies. Phagocytic activity, mixed lymphocyte reaction (MLR and cytokine production were also evaluated. At the end of culturing period, significantly fully matured DCs with large amount cytoplasm and copious dendritic projections were found in the presence of MCM, TNF-α with or without FCM, TCCM, FCM as well as TCCM. Flow cytometric analysis revealed that expression of CD14 decreased in particular in treated DCs, at the 5th day and expression of CD80, CD86 and HLA-DR was higher when FCM, TCCM, FCM plus TCCM were added to maturation factor. This study demonstrated that DCs matured with these methods had optimum function in comparison with either factor alone.

Ultrastructural Histopathology of Vervet Monkey Colonic Epithelium After In Vitro Exposure to Cell-free Supernatants of Shigella Cultures

OpenAIRE

Hill, R. R.; Collins, N. E.; Cowley, H. M.

2011-01-01

The full dysentery syndrome of human shigellosis is often preceded by a transient diarrhoea that may be induced by bacterial extracellular products before invasion of the colonic mucosa and development of subsequent pathology. To examine this hypothesis, we studied the effects of cell-free cultures of Shigella sp. on the ultrastructure of monkey colonic epithelium in vitro. Clinical isolates of shigella strains were grown in a niche-simulating medium. Sheets of colon wall collected from verve...

Supernatants from culture of type I collagen-stimulated PBMC from patients with cutaneous systemic sclerosis versus localized scleroderma demonstrate suppression of MMP-1 by fibroblasts.

Science.gov (United States)

Brown, Monica; Postlethwaite, Arnold E; Myers, Linda K; Hasty, Karen A

2012-06-01

Systemic sclerosis (SSc) is a chronic fibrosing disease characterized by vasculopathy, autoimmunity, and an accumulation of collagen in tissues. Numerous studies have shown that compared to healthy or diseased controls, the peripheral blood mononuclear cells (PBMC) from patients with SSc produce a variety of cytokines or proliferate when cultured with solubilized type I collagen (CI) or constituent α1(II) and α2(I) polypeptide chains. The purpose of this study was to determine whether PBMC isolated from patients with SSc and cultured in vitro with soluble CI elaborated soluble mediators that inhibit the production of collagenase (i.e., matrix metalloproteinase, MMP-1) by fibroblasts. Supernatants of CI-stimulated PBMC from juvenile and adult diffuse cutaneous (dc)SSc patients significantly reduced MMP-1 production by SSc dermal fibroblasts, while supernatants of CI-stimulated PBMC from patients with localized scleroderma (LS) did not. CI-stimulated PBMC culture supernatants from patients with dcSSc in contrast to patients with LS exhibited increased levels of platelet-derived growth factor (PDGF)-AA, PDGF-BB, TNF-α, IL-13, and EGF. Prolonged culture of SSc dermal fibroblasts with recombinant PDGF-BB or IL-13 inhibited the induction of MMP-1 in response to subsequent TNF-α stimulation. These data suggest that therapies aimed at reducing these cytokines may decrease collagen accumulation in SSc, preventing the development of chronic fibrosis.

Distribution of non-LEE-encoded type 3 secretion system dependent effectors in enteropathogenic Escherichia coli

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Fábia A. Salvador

2014-09-01

Full Text Available Enteropathogenic Escherichia coli (EPEC are important human gastroenteritis agents. The prevalence of six non-LEE genes encoding type 3 translocated effectors was investigated. The nleC, cif and nleB genes were more prevalent in typical than in atypical EPEC, although a higher diversity of genes combinations was observed in atypical EPEC.

Antifungal performance of extracellular chitinases and culture supernatants of Streptomyces galilaeus CFFSUR-B12 against Mycosphaerella fijiensis Morelet.

Science.gov (United States)

Castillo, Benjamín Moreno; Dunn, Michael F; Navarro, Karina Guillén; Meléndez, Francisco Holguín; Ortiz, Magdalena Hernández; Guevara, Sergio Encarnación; Palacios, Graciela Huerta

2016-03-01

The tropical and mycoparasite strain Streptomyces galilaeus CFFSUR-B12 was evaluated as an antagonist of Mycosphaerella fijiensis Morelet, causal agent of the Black Sigatoka Disease (BSD) of banana. On zymograms of CFFSUR-B12 culture supernatants, we detected four chitinases of approximately 32 kDa (Chi32), 20 kDa (Chi20), and two with masses well over 170 kDa (ChiU) that showed little migration during denaturing electrophoresis at different concentrations of polyacrylamide. The thymol-sulphuric acid assay showed that the ChiU were glycosylated chitinases. Moreover, matrix assisted laser desorption ionization time-of-flight MS analysis revealed that the ChiU are the same protein and identical to a family 18 chitinase from Streptomyces sp. S4 (gi|498328075). Chi32 was similar to an extracellular protein from Streptomyces albus J1074 (gi|478687481) and Chi20 was non-significantly similar to chitinases from five different strains of Streptomyces (P > 0.05). Subsequently, Chi32 and Chi20 were partially purified by anion exchange and hydrophobic interaction chromatography and tested against M. fijiensis. Chitinases failed to inhibit ascospore germination, but inhibited up to 35 and 62% of germ tube elongation and mycelial growth, respectively. We found that crude culture supernatant and living cells of S. galilaeus CFFSUR-B12 were the most effective in inhibiting M. fijiensis and are potential biocontrol agents of BSD.

SalB inactivation modulates culture supernatant exoproteins and affects autolysis and viability in Enterococcus faecalis OG1RF.

Science.gov (United States)

Shankar, Jayendra; Walker, Rachel G; Wilkinson, Mark C; Ward, Deborah; Horsburgh, Malcolm J

2012-07-01

The culture supernatant fraction of an Enterococcus faecalis gelE mutant of strain OG1RF contained elevated levels of the secreted antigen SalB. Using differential fluorescence gel electrophoresis (DIGE) the salB mutant was shown to possess a unique complement of exoproteins. Differentially abundant exoproteins were identified using matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry. Stress-related proteins including DnaK, Dps family protein, SOD, and NADH peroxidase were present in greater quantity in the OG1RF salB mutant culture supernatant. Moreover, several proteins involved in cell wall synthesis and cell division, including d-Ala-d-Lac ligase and EzrA, were present in reduced quantity in OG1RF salB relative to the parent strain. The salB mutant displayed reduced viability and anomalous cell division, and these phenotypes were exacerbated in a gelE salB double mutant. An epistatic relationship between gelE and salB was not identified with respect to increased autolysis and cell morphological changes observed in the salB mutant. SalB was purified as a six-histidine-tagged protein to investigate peptidoglycan hydrolytic activity; however, activity was not evident. High-pressure liquid chromatography (HPLC) analysis of reduced muropeptides from peptidoglycan digested with mutanolysin revealed that the salB mutant and OG1RF were indistinguishable.

Analysis of Induced Gas Releases During Retrieval of Hanford Double-Shell Tank Waste

International Nuclear Information System (INIS)

Wells, Beric E.; Cuta, Judith M.; Hartley, Stacey A.; Mahoney, Lenna A.; Meyer, Perry A.; Stewart, Charles W.

2002-01-01

Radioactive waste is scheduled to be retrieved from Hanford double-shell tanks AN-103, AN-104, AN-105, and AW-101 to the vitrification plant beginning about 2009. Retrieval may involve decanting the supernatant liquid and/or mixing the waste with jet pumps. In these four tanks, which contain relatively large volumes of retained gas, both of these operations are expected to induce buoyant displacement gas releases that can potentially raise the tank headspace hydrogen concentration to very near the lower flammability limit. This report describes the theory and detailed physical models for both the supernatant decant and jet mixing processes and presents the results from applying the models to these operations in the four tanks. The technical bases for input parameter distributions are elucidated

Validity of antibodies in lymphocyte supernatant in diagnosing tuberculosis in severely malnourished children presenting with pneumonia.

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Mohammod Jobayer Chisti

Full Text Available The diagnosis of tuberculosis (TB in young children can be challenging, especially in severely malnourished children. There is a critical need for improved diagnostics for children. Thus, we sought to evaluate the performance of a technique that measures antibodies in lymphocyte supernatant (ALS for the diagnosis of TB in severely malnourished children presenting with suspected pneumonia.Children less than 5 years with severe acute malnutrition and radiological features of pneumonia admitted to the Dhaka Hospital of International Centre for Diarrhoeal Disease Research, Bangladesh, were enrolled consecutively following informed written consent. In addition to clinical and radiological assessment, samples taken for TB diagnosis included gastric lavage fluid and induced sputum for microbiological confirmation. ALS was measured from venous blood, and results were evaluated in children classified as "confirmed", "non-confirmed TB" or "not TB".Among 224 children who had ALS analysis, 12 (5.4% children had microbiologically "confirmed TB", a further 41 (18% had clinically diagnosed "non-confirmed TB" and the remaining 168 (75% were considered not to have TB. ALS was positive in 89 (40% and negative in 85 (39% of children, with a large number (47 or 21% reported as "borderline". These proportions were similar between the three diagnostic groups. The sensitivity and specificity of ALS when comparing "Confirmed TB" to "Not TB" was only 67% (95% CI: 31-91% and 51% (95% CI: 42-60%, respectively.Our data suggest that ALS is not sufficiently accurate to improve the diagnosis of TB in children with severe malnutrition.

Precompound decay models for medium energy nuclear reactions

International Nuclear Information System (INIS)

Blann, M.

1989-11-01

The formulations used for precompound decay models are presented and explained in terms of the physics of the intranuclear cascade model. Several features of spectra of medium energy (10--1000 MeV) reactions are summarized. Results of precompound plus evaporation calculations from the code ALICE are compared with a wide body of proton, alpha, and heavy ion induced reaction data to illustrate both the power and deficiencies of predicting yield of these reactions in the medium energy regime. 23 refs., 13 figs

Batch test equilibration studies examining the removal of Cs, Sr, and Tc from supernatants from ORNL underground storage tanks by selected ion exchangers

International Nuclear Information System (INIS)

Collins, J.L.; Egan, B.Z.; Anderson, K.K.; Chase, C.W.; Bell, J.T.

1995-01-01

Bench-scale batch equilibration tests have been conducted with supernatants from two underground tanks at the Melton Valley Storage Tank (MVST) Facility at Oak Ridge National Laboratory (ORNL) to determine the effectiveness of selected ion exchangers in removing cesium, strontium, and technetium. Seven sorbents were evaluated for cesium removal, nine for strontium removal, and four for technetium removal. The results indicate that granular potassium cobalt hexacyanoferrate was the most effective of the exchangers evaluated for removing cesium from the supernatants. The powdered forms of sodium titanate (NaTiO) and cystalline silicotitanate (CST) were superior in removing the strontium; however, for the sorbents of suitable particle size for column use, titanium monohydrogen phosphate (TiHP φ), sodium titanate/polyacrylonitrile (NaTiO-PAN), and titanium monohydrogen phosphate/polyacrylonitrile (TiP-PAN) gave the best results and were about equally effective. Reillex trademark 402 was the most effective exchanger in removing the technetium; however, it was only slightly more satisfactory than Reillex trademark HPQ

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

Science.gov (United States)

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

2016-01-01

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

HIV-1-infected macrophages induce astrogliosis by SDF-1α and matrix metalloproteinases

International Nuclear Information System (INIS)

Okamoto, Mika; Wang, Xin; Baba, Masanori

2005-01-01

Brain macrophages/microglia and astrocytes are known to be involved in the pathogenesis of HIV-1-associated dementia (HAD). To clarify their interaction and contribution to the pathogenesis, HIV-1-infected or uninfected macrophages were used as a model of brain macrophages/microglia, and their effects on human astrocytes in vitro were examined. The culture supernatants of HIV-1-infected or uninfected macrophages induced significant astrocyte proliferation, which was annihilated with a neutralizing antibody to stromal cell-derived factor (SDF)-1α or a matrix metalloproteinase (MMP) inhibitor. In these astrocytes, CXCR4, MMP, and tissue inhibitors of matrix metalloproteinase mRNA expression and SDF-1α production were significantly up-regulated. The supernatants of infected macrophages were always more effective than those of uninfected cells. Moreover, the enhanced production of SDF-1α was suppressed by the MMP inhibitor. These results indicate that the activated and HIV-1-infected macrophages can indirectly induce astrocyte proliferation through up-regulating SDF-1α and MMP production, which implies a mechanism of astrogliosis in HAD

Medium-chain triglyceride ameliorates insulin resistance and inflammation in high fat diet-induced obese mice.

Science.gov (United States)

Geng, Shanshan; Zhu, Weiwei; Xie, Chunfeng; Li, Xiaoting; Wu, Jieshu; Liang, Zhaofeng; Xie, Wei; Zhu, Jianyun; Huang, Cong; Zhu, Mingming; Wu, Rui; Zhong, Caiyun

2016-04-01

The aim of the present study was to investigate the in vivo effects of dietary medium-chain triglyceride (MCT) on inflammation and insulin resistance as well as the underlying potential molecular mechanisms in high fat diet-induced obese mice. Male C57BL/6J mice (n = 24) were fed one of the following three diets for a period of 12 weeks: (1) a modified AIN-76 diet with 5 % corn oil (normal diet); (2) a high-fat control diet (17 % w/w lard and 3 % w/w corn oil, HFC); (3) an isocaloric high-fat diet supplemented with MCT (17 % w/w MCT and 3 % w/w corn oil, HF-MCT). Glucose metabolism was evaluated by fasting blood glucose levels and intraperitoneal glucose tolerance test. Insulin sensitivity was evaluated by fasting serum insulin levels and the index of homeostasis model assessment-insulin resistance. The levels of serum interleukin-6 (IL-6), interleukin-10 (IL-10), and tumor necrosis factor-α were measured by ELISA, and hepatic activation of nuclear factor κB (NF-κB) and mitogen-activated protein kinase (MAPK) pathways was determined using western blot analysis. Compared to HFC diet, consumption of HF-MCT did not induce body weight gain and white adipose tissue accumulation in mice. HFC-induced increases in serum fasting glucose and insulin levels as well as glucose intolerance were prevented by HF-MCT diet. Meanwhile, HF-MCT resulted in significantly lower serum IL-6 level and higher IL-10 level, and lower expression levels of inducible nitric oxide synthase and cyclooxygenase-2 protein in liver tissues when compared to HFC. In addition, HF-MCT attenuated HFC-triggered hepatic activation of NF-κB and p38 MAPK. Our study demonstrated that MCT was efficacious in suppressing body fat accumulation, insulin resistance, inflammatory response, and NF-κB and p38 MAPK activation in high fat diet-fed mice. These data suggest that MCT may exert beneficial effects against high fat diet-induced insulin resistance and inflammation.

Lactobacillus reuteri Inhibition of Enteropathogenic Escherichia coli Adherence to Human Intestinal Epithelium

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Alistair eWalsham

2016-03-01

Full Text Available Enteropathogenic E. coli (EPEC is a major cause of diarrheal infant death in developing countries, and probiotic bacteria have been shown to provide health benefits in gastrointestinal infections. In this study, we have investigated the influence of the gut symbiont Lactobacillus reuteri on EPEC adherence to the human intestinal epithelium. Different host cell model systems including non-mucus-producing HT-29 and mucus-producing LS174T intestinal epithelial cell lines as well as human small intestinal biopsies were used. Adherence of L. reuteri to HT-29 cells was strain-specific, and the mucus-binding proteins CmbA and MUB increased binding to both HT-29 and LS174T cells. L. reuteri ATCC PTA 6475 and ATCC 53608 significantly inhibited EPEC binding to HT-29 but not LS174T cells. While pre-incubation of LS174T cells with ATCC PTA 6475 did not affect EPEC A/E lesion formation, it increased the size of EPEC microcolonies. ATCC PTA 6475 and ATCC 53608 binding to the mucus layer resulted in decreased EPEC adherence to small intestinal biopsy epithelium. Our findings show that L. reuteri reduction of EPEC adhesion is strain-specific and has the potential to target either the epithelium or the mucus layer, providing further rationale for the selection of probiotic strains.

Lactobacillus reuteri Inhibition of Enteropathogenic Escherichia coli Adherence to Human Intestinal Epithelium.

Science.gov (United States)

Walsham, Alistair D S; MacKenzie, Donald A; Cook, Vivienne; Wemyss-Holden, Simon; Hews, Claire L; Juge, Nathalie; Schüller, Stephanie

2016-01-01

Enteropathogenic Escherichia coli (EPEC) is a major cause of diarrheal infant death in developing countries, and probiotic bacteria have been shown to provide health benefits in gastrointestinal infections. In this study, we have investigated the influence of the gut symbiont Lactobacillus reuteri on EPEC adherence to the human intestinal epithelium. Different host cell model systems including non-mucus-producing HT-29 and mucus-producing LS174T intestinal epithelial cell lines as well as human small intestinal biopsies were used. Adherence of L. reuteri to HT-29 cells was strain-specific, and the mucus-binding proteins CmbA and MUB increased binding to both HT-29 and LS174T cells. L. reuteri ATCC PTA 6475 and ATCC 53608 significantly inhibited EPEC binding to HT-29 but not LS174T cells. While pre-incubation of LS174T cells with ATCC PTA 6475 did not affect EPEC attaching/effacing (A/E) lesion formation, it increased the size of EPEC microcolonies. ATCC PTA 6475 and ATCC 53608 binding to the mucus layer resulted in decreased EPEC adherence to small intestinal biopsy epithelium. Our findings show that L. reuteri reduction of EPEC adhesion is strain-specific and has the potential to target either the epithelium or the mucus layer, providing further rationale for the selection of probiotic strains.

Optimization of marine waste based-growth media for microbial lipase production using mixture design methodology.

Science.gov (United States)

Sellami, Mohamed; Kedachi, Samiha; Frikha, Fakher; Miled, Nabil; Ben Rebah, Faouzi

2013-01-01

Lipase production by Staphylococcus xylosus and Rhizopus oryzae was investigated using a culture medium based on a mixture of synthetic medium and supernatants generated from tuna by-products and Ulva rigida biomass. The proportion of the three medium components was optimized using the simplex-centroid mixture design method (SCMD). Results indicated that the experimental data were in good agreement with predicted values, indicating that SCMD was a reliable method for determining the optimum mixture proportion of the growth medium. Maximal lipase activities of 12.5 and 23.5 IU/mL were obtained with a 50:50 (v:v) mixture of synthetic medium and tuna by-product supernatant for Staphylococcus xylosus and Rhizopus oryzae, respectively. The predicted responses from these mixture proportions were also validated experimentally.

Enhanced cocaine-induced locomotor sensitization and intrinsic excitability of NAc medium spiny neurons in adult but not adolescent rats susceptible to diet-induced obesity

Science.gov (United States)

Oginsky, Max F.; Maust, Joel D.; Corthell, John T.; Ferrario, Carrie R.

2015-01-01

Rationale Basal and diet-induced differences in mesolimbic function, particularly within the nucleus accumbens (NAc), may contribute to human obesity; these differences may be more pronounced in susceptible populations. Objectives We determined whether there are differences in cocaine-induced behavioral plasticity in rats that are susceptible vs. resistant to diet-induced obesity, and basal differences in the striatal neuron function in adult and adolescent obesity-prone and obesity-resistant rats. Methods Susceptible and resistant outbred rats were identified based on “junk-food” diet-induced obesity. Then, the induction and expression of cocaine-induced locomotor sensitization, which is mediated by enhanced striatal function and is associated with increased motivation for rewards and reward-paired cues, were evaluated. Basal differences in mesolimbic function were examined in selectively bred obesity-prone and obesity-resistant rats (P70-80 and P30-40) using both cocaine induced locomotion and whole-cell patch clamping approaches in NAc core medium spiny neurons (MSNs). Results In rats that became obese after eating “junk-food”, the expression of locomotor sensitization was enhanced compared to non-obese rats, with similarly strong responses to 7.5 and 15 mg/kg cocaine. Without diet manipulation, obesity-prone rats were hyper-responsive to the acute locomotor-activating effects of cocaine, and the intrinsic excitability of NAc core MSNs was enhanced by ~60% at positive and negative potentials. These differences were present in adult, but not adolescent rats. Post-synaptic glutamatergic transmission was similar between groups. Conclusions Mesolimbic systems, particularly NAc MSNs, are hyper-responsive in obesity-prone individuals; and interactions between predisposition and experience influence neurobehavioral plasticity in ways that may promote weight gain and hamper weight loss in susceptible rats. PMID:26612617

Metabolomics Investigation of an Association of Induced Features and Corresponding Fungus during the Co-culture of Trametes versicolor and Ganoderma applanatum

Science.gov (United States)

Xu, Xiao-Yan; Shen, Xiao-Ting; Yuan, Xiao-Jie; Zhou, Yuan-Ming; Fan, Huan; Zhu, Li-Ping; Du, Feng-Yu; Sadilek, Martin; Yang, Jie; Qiao, Bin; Yang, Song

2018-01-01

The co-culture of Trametes versicolor and Ganoderma applanatum is a model of intense basidiomycete interaction, which induces many newly synthesized or highly produced features. Currently, one of the major challenges is an identification of the origin of induced features during the co-culture. Herein, we report a 13C-dynamic labeling analysis used to determine an association of induced features and corresponding fungus even if the identities of metabolites were not available or almost nothing was known of biochemical aspects. After the co-culture of T. versicolor and G. applanatum for 10 days, the mycelium pellets of T. versicolor and G. applanatum were sterilely harvested and then mono-cultured in the liquid medium containing half fresh medium with 13C-labeled glucose as carbon source and half co-cultured supernatants collected on day 10. 13C-labeled metabolome analyzed by LC-MS revealed that 31 induced features including 3-phenyllactic acid and orsellinic acid were isotopically labeled in the mono-culture after the co-culture stimulation. Twenty features were derived from T. versicolor, 6 from G. applanatum, and 5 features were synthesized by both T. versicolor and G. applanatum. 13C-labeling further suggested that 12 features such as previously identified novel xyloside [N-(4-methoxyphenyl)formamide 2-O-beta-D-xyloside] were likely induced through the direct physical interaction of mycelia. Use of molecular network analysis combined with 13C-labeling provided an insight into the link between the generation of structural analogs and producing fungus. Compound 1 with m/z 309.0757, increased 15.4-fold in the co-culture and observed 13C incorporation in the mono-culture of both T. versicolor and G. applanatum, was purified and identified as a phenyl polyketide, 2,5,6-trihydroxy-4, 6-diphenylcyclohex-4-ene-1,3-dione. The biological activity study indicated that this compound has a potential to inhibit cell viability of leukemic cell line U937. The current work sets

Cigarette smoke modulates expression of human rhinovirus-induced airway epithelial host defense genes.

Directory of Open Access Journals (Sweden)

David Proud

Full Text Available Human rhinovirus (HRV infections trigger acute exacerbations of chronic obstructive pulmonary disease (COPD and asthma. The human airway epithelial cell is the primary site of HRV infection and responds to infection with altered expression of multiple genes, the products of which could regulate the outcome to infection. Cigarette smoking aggravates asthma symptoms, and is also the predominant risk factor for the development and progression of COPD. We, therefore, examined whether cigarette smoke extract (CSE modulates viral responses by altering HRV-induced epithelial gene expression. Primary cultures of human bronchial epithelial cells were exposed to medium alone, CSE alone, purified HRV-16 alone or to HRV-16+ CSE. After 24 h, supernatants were collected and total cellular RNA was isolated. Gene array analysis was performed to examine mRNA expression. Additional experiments, using real-time RT-PCR, ELISA and/or western blotting, validated altered expression of selected gene products. CSE and HRV-16 each induced groups of genes that were largely independent of each other. When compared to gene expression in response to CSE alone, cells treated with HRV+CSE showed no obvious differences in CSE-induced gene expression. By contrast, compared to gene induction in response to HRV-16 alone, cells exposed to HRV+CSE showed marked suppression of expression of a number of HRV-induced genes associated with various functions, including antiviral defenses, inflammation, viral signaling and airway remodeling. These changes were not associated with altered expression of type I or type III interferons. Thus, CSE alters epithelial responses to HRV infection in a manner that may negatively impact antiviral and host defense outcomes.

Dissolved oxygen concentration in the medium during cell culture: Defects and improvements.

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Zhang, Kuan; Zhao, Tong; Huang, Xin; He, Yunlin; Zhou, Yanzhao; Wu, Liying; Wu, Kuiwu; Fan, Ming; Zhu, Lingling

2016-03-01

In vitro cell culture has provided a useful model to study the effects of oxygen on cellular behavior. However, it remains unknown whether the in vitro operations themselves affect the medium oxygen levels and the living states of cells. In addition, a prevailing controversy is whether reactive oxygen species (ROS) production is induced by continuous hypoxia or reoxygenation. In this study, we have measured the effects of different types of cell culture containers and the oxygen environment where medium replacement takes place on the actual oxygen tension in the medium. We found that the deviations of oxygen concentrations in the medium are much greater in 25-cm(2) flasks than in 24-well plates and 35-mm dishes. The dissolved oxygen concentrations in the medium were increased after medium replacement in normoxia, but remained unchanged in glove boxes in which the oxygen tension remained at a low level (11.4, 5.7, and 0.5% O2 ). We also found that medium replacement in normoxia increased the number of ROS-positive cells and reduced the cell viability; meanwhile, medium replacement in a glove box did not produce the above effects. Therefore, we conclude that the use of 25-cm(2) flasks should be avoided and demonstrate that continuous hypoxia does not produce ROS, whereas the reoxygenation that occurs during the harvesting of cells leads to ROS and induces cell death. © 2015 International Federation for Cell Biology.

Enteropathogenic Escherichia coli Uses NleA to Inhibit NLRP3 Inflammasome Activation.

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Hilo Yen

2015-09-01

Full Text Available Enteropathogenic and enterohemorrhagic Escherichia coli (EPEC and EHEC are related strains capable of inducing severe gastrointestinal disease. For optimal infection, these pathogens actively modulate cellular functions through the deployment of effector proteins in a type three secretion system (T3SS-dependent manner. In response to enteric pathogen invasion, the Nod-like receptor pyrin domain containing (NLRP inflammasome has been increasingly recognized as an important cytoplasmic sensor against microbial infection by activating caspase-1 and releasing IL-1β. EPEC and EHEC are known to elicit inflammasome activation in macrophages and epithelial cells; however, whether the pathogens actively counteract such innate immune responses is unknown. Using a series of compound effector-gene deletion strains of EPEC, we screened and identified NleA, which could subdue host IL-1β secretion. It was found that the reduction is not because of blocked NF-κB activity; instead, the reduction results from inhibited caspase-1 activation by NleA. Immunostaining of human macrophage-like cells following infection revealed limited formation of inflammasome foci with constituents of total caspase-1, ASC and NLRP3 in the presence of NleA. Pulldown of PMA-induced differentiated THP-1 lysate with purified MBP-NleA reveals that NLRP3 is a target of NleA. The interaction was verified by an immunoprecipitation assay and direct interaction assay in which purified MBP-NleA and GST-NLRP3 were used. We further showed that the effector interacts with regions of NLRP3 containing the PYD and LRR domains. Additionally, NleA was found to associate with non-ubiquitinated and ubiquitinated NLRP3 and to interrupt de-ubiquitination of NLRP3, which is a required process for inflammasome activation. Cumulatively, our findings provide the first example of EPEC-mediated suppression of inflammasome activity in which NieA plays a novel role in controlling the host immune response

Arterial and venous blood pressure and blood flow following femoral angiography with a new non-ionic contrast medium

International Nuclear Information System (INIS)

Nyman, U.; Almen, T.

1978-01-01

At femoral angiography in dogs the effects of a new non-ionic contrast medium (C29) were compared with those of one non-ionic medium (metrizamide) and one ionic medium (meglumine/sodium diatrizoate) in current use. In the leg subjected to angiography the pressure gradient over the peripheral vessels decreased and the femoral blood flow increased. The changes induced by the ionic medium were significantly greater than those induced by metrizamide and C29, whereas no significant difference between the two non-ionic media was recorded. (Auth.)

Mesenchymal stem cell-conditioned medium prevents radiation-induced liver injury by inhibiting inflammation and protecting sinusoidal endothelial cells

International Nuclear Information System (INIS)

Chen Yixing; Zeng Zhaochong; Sun Jing; Huang Yan; Zhang Zhenyu; Zeng Haiying

2015-01-01

Current management of radiation-induced liver injury is limited. Sinusoidal endothelial cell (SEC) apoptosis and inflammation are considered to be initiating events in hepatic damage. We hypothesized that mesenchymal stem cells (MSCs) possess anti-apoptotic and anti-inflammatory actions during hepatic irradiation, acting via paracrine mechanisms. This study aims to examine whether MSC-derived bioactive components are protective against radiation-induced liver injury in rats. MSC-conditioned medium (MSC-CM) was generated from rat bone marrow–derived MSCs. The effect of MSC-CM on the viability of irradiated SECs was examined by flow cytometric analysis. Activation of the Akt and ERK pathways was analyzed by western blot. MSC-CM was also delivered to Sprague–Dawley rats immediately before receiving liver irradiation, followed by testing for pathological features, changes in serum hyaluronic acid, ALT, and inflammatory cytokine levels, and liver cell apoptosis. MSC-CM enhanced the viability of irradiated SECs in vitro and induced Akt and ERK phosphorylation in these cells. Infusion of MSC-CM immediately before liver irradiation provided a significant anti-apoptotic effect on SECs and improved the histopathological features of injury in the irradiated liver. MSC-CM also reduced the secretion and expression of inflammatory cytokines and increased the expression of anti-inflammatory cytokines. MSC-derived bioactive components could be a novel therapeutic approach for treating radiation-induced liver injury. (author)

The down-regulation of the mitogenic fibrinogen receptor (MFR) in serum-containing medium does not occur in defined medium.

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Levesque, J P; Hatzfeld, A; Domart, I; Hatzfeld, J

1990-02-01

Normal human hemopoietic cells such as early bone marrow progenitors, or lymphoma-derived cell lines such as Raji or JM cells, possess a low-affinity receptor specific for fibrinogen. This receptor triggers a mitogenic effect. It differs from the glycoprotein IIb-IIIa which is involved in fibrinogen-induced platelet aggregation. We demonstrate here that this mitogenic fibrinogen receptor (MFR) can be internalized or reexpressed, depending on culture conditions. Internalization was temperature-dependent. At 37 degrees C in the presence of cycloheximide or actinomycin D, the half-life of cell surface MFRs was 2 h, independent of receptor occupancy. Binding of fibrinogen to the MFR resulted in a down-regulation which was fibrinogen dose-dependent. This occurred in serum-supplemented medium but not in defined medium supplemented with fatty acids. Reexpression of MFRs could be induced in 28 to 42 h by serum removal. The down-regulation of mitogenic receptors in plasma or serum could explain why normal cells do not proliferate in the peripheral blood.

Membrane damage induced in cultured human skin fibroblasts by UVA irradiation

International Nuclear Information System (INIS)

Gaboriau, F.; Morliere, P.; Marquis, I.; Moysan, A.; Geze, M.; Dubertret, L.

1993-01-01

Irradiation of cultured human skin fibroblasts with ultraviolet light from 320 to 400 nm (UVA) leads to a decrease in the membrane fluidity exemplified by an enhanced fluorescence anisotropy of the lipophilic fluorescent probe 1-[4-trimethylamino)-phenyl]-6-phenylhexa-1,3,5-triene. This UVA-induced decrease in fluidity is associated with lactate dehydrogenase leakage in the supernatant. Vitamin E, an inhibitor of lipid peroxidation, exerts a protective effect on both phenomena. Therefore, this UVA-induced damage in membrane properties may be related to lipid peroxidation processes. Moreover, exponentially growing cells are more sensitive to these UVA-induced alterations than confluent cells. (Author)

Optimization of high-rate TN removal in a novel constructed wetland integrated with microelectrolysis system treating high-strength digestate supernatant.

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Guo, Luchen; He, Keli; Wu, Shubiao; Sun, Hao; Wang, Yanfei; Huang, Xu; Dong, Renjie

2016-08-01

The potential of high-rate TN removal in three aerated horizontal subsurface-flow constructed wetlands to treat high-strength anaerobic digestate supernatant was evaluated. Different strategies of intermittent aeration and effluent recirculation were applied to compare their effect on nitrogen depuration performance. Additional glucose supply and iron-activated carbon based post-treatment systems were established and examined, respectively, to further remove nitrate that accumulated in the effluents from aerated wetlands. The results showed that intermittent aeration (1 h on:1 h off) significantly improved nitrification with ammonium removal efficiency of 90% (18.1 g/(m(2)·d)), but limited TN removal efficiency (53%). Even though effluent recirculation (a ratio of 1:1) increased TN removal from 53% to 71%, the effluent nitrate concentration was still high. Additional glucose was used as a post-treatment option and further increased the TN removal to 82%; however, this implementation caused additional organic pollution. Furthermore, the iron-activated carbon system stimulated with a microelectrolysis process achieved greater than 85% effluent nitrate removal and resulted in 86% TN removal. Considering the high TN removal rate, aerated constructed wetlands integrated with a microelectrolysis-driven system show great potential for treating high-strength digestate supernatant. Copyright © 2016 Elsevier Ltd. All rights reserved.

High-harmonic generation in a dense medium

International Nuclear Information System (INIS)

Strelkov, V.V.; Platonenko, V.T.; Becker, A.

2005-01-01

The high-harmonic generation in a plasma or gas under conditions when the single-atom response is affected by neighboring ions or atoms of the medium is studied theoretically. We solve numerically the three-dimensional Schroedinger equation for a single-electron atom in the combined fields of the neighboring particles and the laser, and average the results over different random positions of the particles using the Monte Carlo method. Harmonic spectra are calculated for different medium densities and laser intensities. We observe a change of the harmonic properties due to a random variation of the harmonic phase induced by the field of the medium, when the medium density exceeds a certain transition density. The transition density is found to depend on the harmonic order, but it is almost independent of the fundamental intensity. It also differs for the two (shorter and longer) quantum paths. The latter effect leads for ionic densities in the transition regime to a narrowing of the harmonic lines and a shortening of the attosecond pulses generated using a group of harmonics

Multipole electromagnetic moments of neutrino in dispersive medium

International Nuclear Information System (INIS)

Semikov, V.B.; Smorodinskij, Ya.A.; Gosudarstvennyj Komitet po Ispol'zovaniyu Atomnoj Ehnergii SSSR, Moscow

1989-01-01

Four multipole moments for a Dirac and Majorana neutrino in a dispersive medium are calculated viz., the electric monopole (charge), electric dipole, magnetic dipole and anapole dipole moment. For comparison the same quantities are presented in the case of vacuum. The neutrino does not possess an (induced) anapole moment in an isotropic medium; however, in a ferromagnetic such a moment exists and for the Majorana neutrino it is the only electromagnetic cjaracteristic. As an example the cross section for elastic scattering of a Majorana neutrino by nuclei in an isotropic plasma is calculated

Blackbody emission from light interacting with an effective moving dispersive medium.

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Petev, M; Westerberg, N; Moss, D; Rubino, E; Rimoldi, C; Cacciatori, S L; Belgiorno, F; Faccio, D

2013-07-26

Intense laser pulses excite a nonlinear polarization response that may create an effective flowing medium and, under appropriate conditions, a blocking horizon for light. Here, we analyze in detail the interaction of light with such laser-induced flowing media, fully accounting for the medium dispersion properties. An analytical model based on a first Born approximation is found to be in excellent agreement with numerical simulations based on Maxwell's equations and shows that when a blocking horizon is formed, the stimulated medium scatters light with a blackbody emission spectrum. Based on these results, diamond is proposed as a promising candidate medium for future studies of Hawking emission from artificial, dispersive horizons.

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

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Minoru Tomizawa

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

Injection of embryo culture supernatant to the endometrial cavity does not affect outcomes in IVF/ICSI or oocyte donation cycles: a randomized clinical trial.

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Prapas, Yannis; Petousis, Stamatios; Panagiotidis, Yannis; Gullo, Giuseppe; Kasapi, Lia; Papadeothodorou, Achilleas; Prapas, Nikos

2012-06-01

To evaluate whether intrauterine injection of embryo culture supernatant before embryo transfer has any impact on pregnancy and implantation rates. A total of 400 cycles, of which 200 IVF/ICSI and 200 oocyte donor (OD), were randomly assigned to have their uterine cavity injected (group I) or not (group II). Primary endpoints to be studied were pregnancy and implantation rates. Clinical pregnancy rate per transfer (47.87%, 90/188 versus 48.45%, 94/194) based on transvaginal scan findings at 7 weeks of gestation and implantation rate (25.6% versus 26.5%) were similar in the two groups. The day of embryo transfer, day 3 or day 5, did not affect the final outcome. Injection of embryo culture supernatant into the uterine cavity, 30 min before the embryo transfer on either day 3 or 5, neither improves nor adversely affects the pregnancy rate in IVF/ICSI or oocyte donation cycles. Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.

Propionic acid secreted from propionibacteria induces NKG2D ligand expression on human-activated T lymphocytes and cancer cells

DEFF Research Database (Denmark)

Andresen, Lars; Hansen, Karen Aagaard; Jensen, Helle

2009-01-01

We found that propionic acid secreted from propionibacteria induces expression of the NKG2D ligands MICA/B on activated T lymphocytes and different cancer cells, without affecting MICA/B expression on resting peripheral blood cells. Growth supernatant from propionibacteria or propionate alone cou...

Campylobacter jejuni induces diverse kinetics and profiles of cytokine genes in INT-407 cells

International Nuclear Information System (INIS)

Al-Amri, Ahlam I.; Bakhiet, Moiz O.; Botta, Giuseppe A.; Tabbara, Khaled S.; Ismaeel, Abdelrahman Y.; Al-Mahmeed, Ali E.; Bin Danya, Khalid M.

2008-01-01

Objective was to examine the kinetic ability of embryonic human epithelial INT-407 cells to express messenger ribonucleic acid (mRNA) for various cytokines and chemokines in response to Campylobacter jejuni (C. jejuni) stimulation. In an experimental single-blind study, cultured embryonic human epithelial INT-407 cells were treated with different concentrations of viable C. jejuni, its sonicated and filtered supernatant. A modified non-radioactive in situ hybridization using probe cocktails was used to measure mRNA levels for the pro-inflammatory cytokines interleukin (IL)-1beta, IL-6, interferon-gamma, tumor necrosis factor (TNF)-alpha, transforming growth factor (TGF)-beta1 and IL-8 and the anti-inflammatory cytokines, IL-4 and IL-10. The study was carried out from September 2005 to March 2007 at the Department of Microbiology, Immunology and Infectious Diseases, College of Medicine and Medical Sciences, Arabian Gulf University, Bahrain. Viable C. jejuni sonicated bacteria and filtered supernatant induced high mRNA expression for the pro-inflammatory cytokines IL-1 beta, IL-6, IFN-gama, TNF-alpha, TGF-beta and IL-8 which peaked at the 12 hours post stimulation. Anti-inflammatory cytokine IL-4 and IL-10 mNRA expression were induced maximally at 3 hours post stimulation mainly by sonicated bacteria and filtrated supernatant, however, not with living bacteria and filtrated supernatant, however, not with living bacteria. Untreated embryonic human epithelial INT-407 cells expressed low amount of mNRA for the various cytokines and chemokines at all time points. For each cytokine, 4 samples were used per time hour. This study demonstrated that embryonic human epithelial INT-407 cells in response to viable C. jejuni or its cytotxins can alter cytokine and chemokine mNRA expression patterns and kinetics suggesting a potential role for these mediators in the immunopathogenesis of the infection caused by this pathogen, which might be relevant for future immunotherapeutic

Th1-Induced CD106 Expression Mediates Leukocytes Adhesion on Synovial Fibroblasts from Juvenile Idiopathic Arthritis Patients.

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Maggi, Laura; Margheri, Francesca; Luciani, Cristina; Capone, Manuela; Rossi, Maria Caterina; Chillà, Anastasia; Santarlasci, Veronica; Mazzoni, Alessio; Cimaz, Rolando; Liotta, Francesco; Maggi, Enrico; Cosmi, Lorenzo; Del Rosso, Mario; Annunziato, Francesco

2016-01-01

This study tested the hypothesis that subsets of human T helper cells can orchestrate leukocyte adhesion to synovial fibroblasts (SFbs), thus regulating the retention of leukocytes in the joints of juvenile idiopathic arthritis (JIA) patients. Several cell types, such as monocytes/macrophages, granulocytes, T and B lymphocytes, SFbs and osteoclasts participate in joint tissue damage JIA. Among T cells, an enrichment of classic and non-classic Th1 subsets, has been found in JIA synovial fluid (SF), compared to peripheral blood (PB). Moreover, it has been shown that IL-12 in the SF of inflamed joints mediates the shift of Th17 lymphocytes towards the non-classic Th1 subset. Culture supernatants of Th17, classic and non-classic Th1 clones, have been tested for their ability to stimulate proliferation, and to induce expression of adhesion molecules on SFbs, obtained from healthy donors. Culture supernatants of both classic and non-classic Th1, but not of Th17, clones, were able to induce CD106 (VCAM-1) up-regulation on SFbs. This effect, mediated by tumor necrosis factor (TNF)-α, was crucial for the adhesion of circulating leukocytes on SFbs. Finally, we found that SFbs derived from SF of JIA patients expressed higher levels of CD106 than those from healthy donors, resembling the phenotype of SFbs activated in vitro with Th1-clones supernatants. On the basis of these findings, we conclude that classic and non-classic Th1 cells induce CD106 expression on SFbs through TNF-α, an effect that could play a role in leukocytes retention in inflamed joints.

Conversion of Squid Pen to Homogentisic Acid via Paenibacillus sp. TKU036 and the Antioxidant and Anti-Inflammatory Activities of Homogentisic Acid

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San-Lang Wang

2016-10-01

Full Text Available The culture supernatant of Paenibacillus sp. TKU036, a bacterium isolated from Taiwanese soils, showed high antioxidant activity (85% when cultured in a squid pen powder (SPP-containing medium at 37 °C for three days. Homogentisic acid (2,5-dihydroxyphenylacetic acid, HGA was isolated and found to be the major antioxidant in the culture supernatant of the SPP-containing medium fermented by Paenibacillus sp. TKU036. Tryptophan was also present in the culture supernatant. The results of high-performance liquid chromatography (HPLC fingerprinting showed that HGA and tryptophan were produced via fermentation but did not pre-exist in the unfermented SPP-containing medium. Neither HGA nor tryptophan was found in the culture supernatants obtained from the fermentation of nutrient broth or other chitinous material, i.e., medium containing shrimp head powder, by Paenibacillus sp. TKU036. The production of HGA via microorganisms has rarely been reported. In this study, we found that squid pen was a potential carbon and nitrogen source for Paenibacillus sp. Tryptophan (105 mg/L and HGA (60 mg/L were recovered from the culture supernatant. The isolated HGA was found to have higher antioxidant activity (IC50 = 6.9 μg/mL than α-tocopherol (IC50 = 17.6 μg/mL. The anti-inflammatory activity of the isolated HGA (IC50 = 10.14 μg/mL was lower than that of quercetin (IC50 = 1.14 μg/mL. As a result, squid pen, a fishery processing byproduct, is a valuable material for the production of tryptophan and the antioxidant and anti-inflammatory HGA via microbial conversion.

Characterization of a Large Antibiotic Resistance Plasmid Found in Enteropathogenic Escherichia coli Strain B171 and Its Relatedness to Plasmids of Diverse E. coli and Shigella Strains.

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Hazen, Tracy H; Michalski, Jane; Nagaraj, Sushma; Okeke, Iruka N; Rasko, David A

2017-09-01

Enteropathogenic Escherichia coli (EPEC) is a leading cause of severe infantile diarrhea in developing countries. Previous research has focused on the diversity of the EPEC virulence plasmid, whereas less is known regarding the genetic content and distribution of antibiotic resistance plasmids carried by EPEC. A previous study demonstrated that in addition to the virulence plasmid, reference EPEC strain B171 harbors a second, larger plasmid that confers antibiotic resistance. To further understand the genetic diversity and dissemination of antibiotic resistance plasmids among EPEC strains, we describe the complete sequence of an antibiotic resistance plasmid from EPEC strain B171. The resistance plasmid, pB171_90, has a completed sequence length of 90,229 bp, a GC content of 54.55%, and carries protein-encoding genes involved in conjugative transfer, resistance to tetracycline ( tetA ), sulfonamides ( sulI ), and mercury, as well as several virulence-associated genes, including the transcriptional regulator hha and the putative calcium sequestration inhibitor ( csi ). In silico detection of the pB171_90 genes among 4,798 publicly available E. coli genome assemblies indicates that the unique genes of pB171_90 ( csi and traI ) are primarily restricted to genomes identified as EPEC or enterotoxigenic E. coli However, conserved regions of the pB171_90 plasmid containing genes involved in replication, stability, and antibiotic resistance were identified among diverse E. coli pathotypes. Interestingly, pB171_90 also exhibited significant similarity with a sequenced plasmid from Shigella dysenteriae type I. Our findings demonstrate the mosaic nature of EPEC antibiotic resistance plasmids and highlight the need for additional sequence-based characterization of antibiotic resistance plasmids harbored by pathogenic E. coli . Copyright © 2017 American Society for Microbiology.

Hydration for the prevention of contrast medium-induced nephropathy. An update

International Nuclear Information System (INIS)

Heinrich, M.; Uder, M.

2006-01-01

Contrast medium-induced nephropathy (CIN) continues to be one of the most common causes of hospital-acquired acute renal failure. Since most of the clinical studies on the prophylactic use of different drugs to prevent CIN produced disappointing results, hydration remains the mainstay of prophylaxis. A number of recent prospective randomized trials provided further evidence of the effectiveness of hydration and relevant information regarding the optimization of hydration protocols. It was shown that a bolus hydration solely during examination is not sufficient to prevent CIN. In addition, isotonic 0.9% saline was superior to the commonly used halfisotonic 0.45% saline in another trial. An outpatient hydration protocol including oral hydration before the examination followed by forced intravenous hydration over 6 hrs. beginning 30 to 60 min. prior to examination seems to be comparable to the usual hydration over 24 hrs. Another hydration protocol, which could also be very attractive especially for outpatients, included the infusion of sodium bicarbonate. In a recent trial, hydration with sodium bicarbonate, given as a bolus for 1 hr. prior to examination followed by an infusion for 6 hrs. after examination, was more effective than hydration with sodium chloride for the prophylaxis of CIN. However, there is still a lack of large-scale, multi-center trials comparing different hydration protocols and investigating their influence on clinically relevant endpoints such as mortality or the need for dialysis. (orig.)

Lactobacillus rhamnosus GG supernatant enhance neonatal resistance to systemic Escherichia coli K1 infection by accelerating development of intestinal defense

OpenAIRE

Xiaolong He; Qing Zeng; Santhosh Puthiyakunnon; Zhijie Zeng; Weijun Yang; Jiawen Qiu; Lei Du; Swapna Boddu; Tongwei Wu; Danxian Cai; Sheng-He Huang; Hong Cao

2017-01-01

The objective of this study was to determine whether Lactobacillus rhamnosus GG culture supernatant (LCS) has a preventive effect against gut-derived systemic neonatal Escherichia coli (E. coli) K1 infection. The preventive effects were evaluated in human colonic carcinoma cell line Caco-2 and neonatal rat models. Our in vitro results showed that LCS could block adhesion, invasion and translocation of E. coli K1 to Caco-2 monolayer via up-regulating mucin production and maintaining intestinal...

UVB-irradiated keratinocytes induce melanoma-associated ganglioside GD3 synthase gene in melanocytes via secretion of tumor necrosis factor α and interleukin 6

Energy Technology Data Exchange (ETDEWEB)

Miyata, Maiko [Department of Life and Medical Sciences, Chubu University Faculty of Life and Health Sciences, Matsumoto, Kasugai 487-8501 (Japan); Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya 466-0065 (Japan); Ichihara, Masatoshi; Tajima, Orie; Sobue, Sayaka; Kambe, Mariko [Department of Life and Medical Sciences, Chubu University Faculty of Life and Health Sciences, Matsumoto, Kasugai 487-8501 (Japan); Sugiura, Kazumitsu [Department of Dermatology, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya 466-0065 (Japan); Furukawa, Koichi, E-mail: koichi@med.nagoya-u.ac.jp [Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya 466-0065 (Japan); Furukawa, Keiko [Department of Life and Medical Sciences, Chubu University Faculty of Life and Health Sciences, Matsumoto, Kasugai 487-8501 (Japan); Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya 466-0065 (Japan)

2014-03-07

Highlights: • Melanocytes showed low ST8SIA1 and high B3GALT4 levels in contrast with melanomas. • Direct UVB irradiation of melanocytes did not induce ganglioside synthase genes. • Culture supernatants of UVB-irradiated keratinocytes induced ST8SIA1 in melanocytes. • TNFα and IL-6 secreted from keratinocytes enhanced ST8SIA1 expression in melanocytes. • Inflammatory cytokines induced melanoma-related ST8SIA1 in melanocytes. - Abstract: Although expression of gangliosides and their synthetic enzyme genes in malignant melanomas has been well studied, that in normal melanocytes has been scarcely analyzed. In particular, changes in expression levels of glycosyltransferase genes responsible for ganglioside synthesis during evolution of melanomas from melanocytes are very important to understand roles of gangliosides in melanomas. Here, expression of glycosyltransferase genes related to the ganglioside synthesis was analyzed using RNAs from cultured melanocytes and melanoma cell lines. Quantitative RT-PCR revealed that melanomas expressed high levels of mRNA of GD3 synthase and GM2/GD2 synthase genes and low levels of GM1/GD1b synthase genes compared with melanocytes. As a representative exogenous stimulation, effects of ultraviolet B (UVB) on the expression levels of 3 major ganglioside synthase genes in melanocytes were analyzed. Although direct UVB irradiation of melanocytes caused no marked changes, culture supernatants of UVB-irradiated keratinocytes (HaCaT cells) induced definite up-regulation of GD3 synthase and GM2/GD2 synthase genes. Detailed examination of the supernatants revealed that inflammatory cytokines such as TNFα and IL-6 enhanced GD3 synthase gene expression. These results suggest that inflammatory cytokines secreted from UVB-irradiated keratinocytes induced melanoma-associated ganglioside synthase genes, proposing roles of skin microenvironment in the promotion of melanoma-like ganglioside profiles in melanocytes.

UVB-irradiated keratinocytes induce melanoma-associated ganglioside GD3 synthase gene in melanocytes via secretion of tumor necrosis factor α and interleukin 6

International Nuclear Information System (INIS)

Miyata, Maiko; Ichihara, Masatoshi; Tajima, Orie; Sobue, Sayaka; Kambe, Mariko; Sugiura, Kazumitsu; Furukawa, Koichi; Furukawa, Keiko

2014-01-01

Highlights: • Melanocytes showed low ST8SIA1 and high B3GALT4 levels in contrast with melanomas. • Direct UVB irradiation of melanocytes did not induce ganglioside synthase genes. • Culture supernatants of UVB-irradiated keratinocytes induced ST8SIA1 in melanocytes. • TNFα and IL-6 secreted from keratinocytes enhanced ST8SIA1 expression in melanocytes. • Inflammatory cytokines induced melanoma-related ST8SIA1 in melanocytes. - Abstract: Although expression of gangliosides and their synthetic enzyme genes in malignant melanomas has been well studied, that in normal melanocytes has been scarcely analyzed. In particular, changes in expression levels of glycosyltransferase genes responsible for ganglioside synthesis during evolution of melanomas from melanocytes are very important to understand roles of gangliosides in melanomas. Here, expression of glycosyltransferase genes related to the ganglioside synthesis was analyzed using RNAs from cultured melanocytes and melanoma cell lines. Quantitative RT-PCR revealed that melanomas expressed high levels of mRNA of GD3 synthase and GM2/GD2 synthase genes and low levels of GM1/GD1b synthase genes compared with melanocytes. As a representative exogenous stimulation, effects of ultraviolet B (UVB) on the expression levels of 3 major ganglioside synthase genes in melanocytes were analyzed. Although direct UVB irradiation of melanocytes caused no marked changes, culture supernatants of UVB-irradiated keratinocytes (HaCaT cells) induced definite up-regulation of GD3 synthase and GM2/GD2 synthase genes. Detailed examination of the supernatants revealed that inflammatory cytokines such as TNFα and IL-6 enhanced GD3 synthase gene expression. These results suggest that inflammatory cytokines secreted from UVB-irradiated keratinocytes induced melanoma-associated ganglioside synthase genes, proposing roles of skin microenvironment in the promotion of melanoma-like ganglioside profiles in melanocytes

Typing of O26 enterohaemorrhagic and enteropathogenic Escherichia coli isolated from humans and cattle with IS621 multiplex PCR-based fingerprinting.

Science.gov (United States)

Mainil, J G; Bardiau, M; Ooka, T; Ogura, Y; Murase, K; Etoh, Y; Ichihara, S; Horikawa, K; Buvens, G; Piérard, D; Itoh, T; Hayashi, T

2011-09-01

This study evaluated a typing method of O26:H11 enterohaemorrhagic and enteropathogenic Escherichia coli (EHEC and EPEC) based on the variation in genomic location and copy numbers of IS621. Two multiplex PCRs, targeting either the left (5') or right (3') IS/chromosome junction of 12 IS621 insertion sites and one PCR specific of another truncated copy, were developed. Thirty-eight amplification profiles were observed amongst a collection of 69 human and bovine O26:H11 EHEC and EPEC. Seventy-one per cent of the 45 EHEC and EPEC with identical IS621 fingerprints within groups of two, three or four isolates had >85% pulsed field gel electrophoresis (PFGE) profile similarity, including four groups of epidemiologically related EHEC or EPEC, while most of the groups had identical IS621 fingerprints and PFGE profiles. The IS621 fingerprinting and the PFGE are complementary typing assays of EHEC and EPEC; though, the former is less discriminatory. The IS621 printing method represents a rapid (24 h) first-line surveillance and typing assay, to compare and trace back O26:H11 EHEC and EPEC during surveys in farms, multiple human cases and outbreaks. Journal of Applied Microbiology © 2011 The Society for Applied Microbiology. No claim to Belgian Government works.

Isolation and serological identification of enteropathogenic Escherichia coli in pasteurized milk in Brazil

Directory of Open Access Journals (Sweden)

Zoraide N da Silva

2001-08-01

Full Text Available OBJECTIVE: To evaluate the microbiological quality of pasteurized milk commercialized in Rio de Janeiro, Brazil, and determine serologically enteropathogenic Escherichia coli (EPEC strains in E. coli isolates obtained from milk samples. METHODS: Ninety samples of pasteurized milk -- types B and C -- of three different commercial brands, purchased in supermarkets and bakeries in Rio de Janeiro, were examined. The amount of total and fecal coliform bacteria was estimated using the Most Probable Number technique. Mesophilic, psychrotrophic, and thermoduric microorganism counts were determined by the Standard Plate Count technique. Isolation and identification of E. coli were carried out using conventional physiological tests. Commercial antisera were used for serological characterization of EPEC. RESULTS: The three milk brands analyzed revealed bacterial counts above the regulated values of the Brazilian government. It was found that among 208 strains of E. coli isolated, 46 (22.1% were serologically classified as EPEC. The most common EPEC serogroup was O55 (15.2%. CONCLUSIONS: Though recent studies on virulence factors indicate that not all strains serologically classified as EPEC are able to attaching/effacing lesion, it is believed that the isolation of EPEC serogroups from pasteurized milk represent a potential risk for children, as well as an indicative of the presence of other enteropathogens.

Atypical Enteropathogenic Escherichia coli Strains form Biofilm on Abiotic Surfaces Regardless of Their Adherence Pattern on Cultured Epithelial Cells

Directory of Open Access Journals (Sweden)

Hebert F. Culler

2014-01-01

Full Text Available The aim of this study was to determine the capacity of biofilm formation of atypical enteropathogenic Escherichia coli (aEPEC strains on abiotic and biotic surfaces. Ninety-one aEPEC strains, isolated from feces of children with diarrhea, were analyzed by the crystal violet (CV assay on an abiotic surface after 24 h of incubation. aEPEC strains representing each HEp-2 cell type of adherence were analyzed after 24 h and 6, 12, and 18 days of incubation at 37°C on abiotic and cell surfaces by CFU/cm2 counting and confocal laser scanning microscopy (CLSM. Biofilm formation on abiotic surfaces occurred in 55 (60.4% of the aEPEC strains. There was no significant difference in biofilm biomass formation on an abiotic versus prefixed cell surface. The biofilms could be visualized by CLSM at various developmental stages. aEPEC strains are able to form biofilm on an abiotic surface with no association with their adherence pattern on HEp-2 cells with the exception of the strains expressing UND (undetermined adherence. This study revealed the capacity of adhesion and biofilm formation by aEPEC strains on abiotic and biotic surfaces, possibly playing a role in pathogenesis, mainly in cases of persistent diarrhea.

1,25-dihydroxyvitamin D3 induces LL-37 and HBD-2 production in keratinocytes from diabetic foot ulcers promoting wound healing: an in vitro model.

Science.gov (United States)

Gonzalez-Curiel, Irma; Trujillo, Valentin; Montoya-Rosales, Alejandra; Rincon, Kublai; Rivas-Calderon, Bruno; deHaro-Acosta, Jeny; Marin-Luevano, Paulina; Lozano-Lopez, Daniel; Enciso-Moreno, Jose A; Rivas-Santiago, Bruno

2014-01-01

Diabetic foot ulcers (DFU) are one of the most common diabetes-related cause of hospitalization and often lead to severe infections and poor healing. It has been recently reported that patients with DFU have lower levels of antimicrobial peptides (AMPs) at the lesion area, which contributes with the impairment of wound healing. The aim of this study was to determine whether 1,25-dihydroxyvitamin D3 (1,25 (OH)2 D3) and L-isoleucine induced HBD-2 and LL-37 in primary cultures from DFU. We developed primary cell cultures from skin biopsies from 15 patients with DFU and 15 from healthy donors. Cultures were treated with 1,25 (OH)2D3 or L-isoleucine for 18 h. Keratinocytes phenotype was identified by western blot and flow cytometry. Real time qPCR for DEFB4, CAMP and VDR gene expression was performed as well as an ELISA to measure HBD-2 and LL-37 in supernatant. Antimicrobial activity, in vitro, wound healing and proliferation assays were performed with conditioned supernatant. The results show that primary culture from DFU treated with 1,25(OH)2D3, increased DEFB4 and CAMP gene expression and increased the production of HBD-2 and LL-37 in the culture supernatant. These supernatants had antimicrobial activity over E. coli and induced remarkable keratinocyte migration. In conclusion the 1,25(OH)2D3 restored the production of AMPs in primary cell from DFU which were capable to improve the in vitro wound healing assays, suggesting their potential therapeutic use on the treatment of DFU.

Assessment of real-time PCR method for detection of EGFR mutation using both supernatant and cell pellet of malignant pleural effusion samples from non-small-cell lung cancer patients.

Science.gov (United States)

Shin, Saeam; Kim, Juwon; Kim, Yoonjung; Cho, Sun-Mi; Lee, Kyung-A

2017-10-26

EGFR mutation is an emerging biomarker for treatment selection in non-small-cell lung cancer (NSCLC) patients. However, optimal mutation detection is hindered by complications associated with the biopsy procedure, tumor heterogeneity and limited sensitivity of test methodology. In this study, we evaluated the diagnostic utility of real-time PCR using malignant pleural effusion samples. A total of 77 pleural fluid samples from 77 NSCLC patients were tested using the cobas EGFR mutation test (Roche Molecular Systems). Pleural fluid was centrifuged, and separated cell pellets and supernatants were tested in parallel. Results were compared with Sanger sequencing and/or peptide nucleic acid (PNA)-mediated PCR clamping of matched tumor tissue or pleural fluid samples. All samples showed valid real-time PCR results in one or more DNA samples extracted from cell pellets and supernatants. Compared with other molecular methods, the sensitivity of real-time PCR method was 100%. Concordance rate of real-time PCR and Sanger sequencing plus PNA-mediated PCR clamping was 98.7%. We have confirmed that real-time PCR using pleural fluid had a high concordance rate compared to conventional methods, with no failed samples. Our data demonstrated that the parallel real-time PCR testing using supernatant and cell pellet could offer reliable and robust surrogate strategy when tissue is not available.

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

Science.gov (United States)

Sun, Dajun D; Lee, Ping I

2015-08-10

same ASD system induced by changes in the drug release mechanism in dissolution medium of a different pH. Copyright © 2015 Elsevier B.V. All rights reserved.

Blockade by fenspiride of endotoxin-induced neutrophil migration in the rat.

Science.gov (United States)

Cunha, F Q; Boukili, M A; da Motta, J I; Vargaftig, B B; Ferreira, S H

1993-07-06

Fenspiride, an antiinflammatory drug with low anti-cyclooxygenase activity, administered orally at 60-200 mg/kg inhibited neutrophil migration into peritoneal and air pouches cavities as well as exudation into peritoneal cavities induced by endotoxin but not induced by carrageenin. Up to 100 microM, fenspiride failed to inhibit the in vitro release of a neutrophil chemotactic activity by endotoxin-stimulated macrophages and the in vivo migration into the peritoneal cavities induced by the supernatant of those macrophages. The release of tumour necrosis factor by stimulated macrophages was inhibited by fenspiride in a dose-dependent manner. These results suggest that the antiinflammatory effects of fenspiride are associated with the inhibition of the tumour necrosis factor release by resident macrophages.

Propagation of frequency-chirped laser pulses in a medium of atoms with a Λ-level scheme

International Nuclear Information System (INIS)

Demeter, G.; Dzsotjan, D.; Djotyan, G. P.

2007-01-01

We study the propagation of frequency-chirped laser pulses in optically thick media. We consider a medium of atoms with a Λ level-scheme (Lambda atoms) and also, for comparison, a medium of two-level atoms. Frequency-chirped laser pulses that induce adiabatic population transfer between the atomic levels are considered. They induce transitions between the two lower (metastable) levels of the Λ-atoms and between the ground and excited states of the two-level atoms. We show that associated with this adiabatic population transfer in Λ-atoms, there is a regime of enhanced transparency of the medium--the pulses are distorted much less than in the medium of two-level atoms and retain their ability to transfer the atomic population much longer during propagation

Tumour necrosis factor and interleukin-6 production induced by components associated with merozoite proteins of Plasmodium falciparum

DEFF Research Database (Denmark)

Jakobsen, P H; Moon, R; Ridley, R G

1993-01-01

purified from culture supernatants, using immobilized monoclonal antibodies specific for RAP-1 or MSP-1, stimulated normal human mononuclear cells to produce TNF and IL-6 in vitro. However, stimulation of TNF was absent, and that of IL-6 was reduced, when the antigens were purified from detergent extracts...... of infected erythrocytes. These results indicate that the RAP-1 and MSP-1 proteins themselves do not stimulate the production of TNF. Instead, other components associating with these exoantigens may be responsible for the TNF production. Mouse antisera blocking TNF production stimulated by P. yoelii...... exoantigens also blocked TNF production stimulated by material affinity purified from P. falciparum culture supernatants using RAP-1 specific monoclonal antibody, indicating the conserved structure of the TNF inducing component....

Maturation of human dendritic cells by monocyte-conditioned medium is dependent upon trace amounts of lipopolysaccharide inducing tumour necrosis factor alpha

DEFF Research Database (Denmark)

Nersting, Jacob; Svenson, Morten; Andersen, Vagn

2003-01-01

We investigated the ability of monocyte-conditioned medium (MCM), generated by monocytes cultured on plastic-immobilised immunoglobulin, to stimulate maturation of human monocyte-derived dendritic cells (DC). Earlier reports suggest that MCM is a strong inducer of irreversible DC maturation......, whereas we find, that adding a small amount of lipopolysaccharide (LPS) to the MCM-generating cultures is required for the production of a DC-stimulatory MCM. Moreover, compared with addition of LPS directly to the DC cultures, stimulation via MCM cultures increases by several fold the DC...

Effect of Lactobacillus sp. isolates supernatant on Escherichia coli O157:H7 enhances the role of organic acids production as a factor for pathogen control

Directory of Open Access Journals (Sweden)

Larissa B. Poppi

2015-04-01

Full Text Available Many attempts have been made to establish the control of foodborne pathogens through Lactobacillus isolates and their metabolism products with success being obtained in several situations. The aim of this study was to investigate the antagonistic effect of eight Lactobacillus isolates, including L. casei subsp. pseudoplantarum, L. plantarum, L. reuteri and L. delbrueckii subsp. delbrueckii, on the pathogenic Escherichia colistrain O157:H7. The inhibitory effect of pure cultures and two pooled cultures supernatants of Lactobacillus on the growth of pathogenic bacteria was evaluated by the spot agar method and by monitoring turbidity. Antimicrobial activity was confirmed for L. reuteri and L. delbrueckii subsp. delbrueckii and for a pool of lactic acid bacteria. The neutralized supernatant of the pool exerted a higher antimicrobial activity than that of the individual strains. Furthermore, D-lactic acid and acetic acid were produced during growth of the Lactobacillus isolates studied.

Multi-particle correlations and KNO scaling in the medium-induced jet evolution

Energy Technology Data Exchange (ETDEWEB)

Escobedo, Miguel A.; Iancu, Edmond [Institut de physique théorique, Université Paris Saclay, CNRS, CEA,F-91191 Gif-sur-Yvette (France)

2016-12-20

We study the gluon distribution produced via successive medium-induced branchings by an energetic jet propagating through a weakly-coupled quark-gluon plasma. We show that under suitable approximations the evolution of the jet can be described as a classical stochastic process, which is exactly solvable. For this process, we construct exact analytic solutions for all the n-point correlation functions (the n-body densities in the space of energy). The corresponding results for the one-point and the two-point functions were already known, but those for the higher-point functions are new. These results demonstrate strong correlations associated with the existence of common ancestors in the branching process. By integrating these n-point functions over the gluon energies, we deduce the mean gluon multiplicity 〈N〉 as well as the higher moments 〈N{sup p}〉 with p≥2. We find that the multiplicities of the soft gluons are parametrically large and show a remarkable regularity, known as Koba-Nielsen-Olesen (KNO) scaling: the reduced moments 〈N{sup p}〉/〈N〉{sup p} are pure numbers, independent of any of the physical parameters of the problem. We recognize a special negative binomial distribution which is characterized by large statistical fluctuations. These predictions can be tested in Pb+Pb collisions at the LHC, via event-by-event measurements of the di-jet asymmetry.

Oral contrast medium in PET/CT: should you or shouldn't you?

International Nuclear Information System (INIS)

Groves, Ashley M.; Kayani, Irfan; Dickson, John C.; Townsend, Caroline; Croasdale, Ian; Syed, Rizwan; Nagabushan, Nagesh; Hain, Sharon F.; Ell, Peter J.; Bomanji, Jamshed B.

2005-01-01

It has been suggested that the use of computed tomography (CT) positive contrast agents has led to attenuation-induced artefacts on 18 F-fluorodeoxyglucose positron emission tomography ( 18 F-FDG PET/CT) systems. Consequently, centres may withhold the use of such agents. Whilst there is theoretical evidence to support the aforementioned claim, the clinical relevance of the induced artefacts has not been widely established. Moreover, the potential benefits of bowel enhancement on PET/CT have yet to be formally evaluated. We therefore prospectively examined PET/CT studies to assess whether the use of oral contrast medium induces clinically relevant artefacts and whether the use of these agents is diagnostically helpful. Over a 2-month period, 18 F-FDG PET/CT images were prospectively reviewed from 200 patients following Gastrografin administration 2 h prior to examination. Both a radiologist and a nuclear medicine physician reviewed the images for contrast medium-mediated clinically relevant artefacts. Artefacts were sought on the CT attenuation-corrected images and were compared with the appearance on non-attenuated-corrected images. The number of examinations in which the oral contrast aided image interpretation was also noted. There were no oral contrast medium-induced clinically significant artefacts. In 38 of the 200 patients, oral contrast aided image interpretation (owing to differentiation of mass/node from bowel, discrimination of intestinal wall from lumen or definition of the anatomy of a relevant site). In 33 of these 38 patients, the anatomical site of interest was the abdomen/pelvis. The use of oral contrast medium in 18 F-FDG PET studies should not be withheld as it improves image interpretation and does not produce clinically significant artefacts. (orig.)

Evaluation of the effects of a plasma activated medium on cancer cells

Energy Technology Data Exchange (ETDEWEB)

Mohades, S.; Laroussi, M., E-mail: mlarouss@odu.edu; Sears, J.; Barekzi, N.; Razavi, H. [Plasma Engineering and Medicine Institute, Old Dominion University, Norfolk, Virginia 23529 (United States)

2015-12-15

The interaction of low temperature plasma with liquids is a relevant topic of study to the field of plasma medicine. This is because cells and tissues are normally surrounded or covered by biological fluids. Therefore, the chemistry induced by the plasma in the aqueous state becomes crucial and usually dictates the biological outcomes. This process became even more important after the discovery that plasma activated media can be useful in killing various cancer cell lines. Here, we report on the measurements of concentrations of hydrogen peroxide, a species known to have strong biological effects, produced by application of plasma to a minimum essential culture medium. The activated medium is then used to treat SCaBER cancer cells. Results indicate that the plasma activated medium can kill the cancer cells in a dose dependent manner, retain its killing effect for several hours, and is as effective as apoptosis inducing drugs.

Proteomic analysis of post-nuclear supernatant fraction and percoll-purified membranes prepared from brain cortex of rats exposed to increasing doses of morphine

Czech Academy of Sciences Publication Activity Database

Ujčíková, Hana; Eckhardt, Adam; Kagan, Dmytro; Roubalová, Lenka; Svoboda, Petr

2014-01-01

Roč. 12, Feb 14 (2014), s. 11 ISSN 1477-5956 R&D Projects: GA ČR(CZ) GAP207/12/0919; GA ČR(CZ) GBP304/12/G069 Institutional support: RVO:67985823 Keywords : morphine * long-term exposure * rat brain cortex * isolated plasma membranes * post-nuclear supernatant * 2D electrophoresis Subject RIV: CE - Biochemistry Impact factor: 1.725, year: 2014

Effect of bacterial components of mixed culture supernatants of planktonic and biofilm Pseudomonas aeruginosa with commensal Escherichia coli on the neutrophil response in vitro.

Science.gov (United States)

Maslennikova, Irina L; Kuznetsova, Marina V; Nekrasova, Irina V; Shirshev, Sergei V

2017-11-30

Pseudomonas aeruginosa (PA) responsible for acute and chronic infections often forms a well-organized bacterial population with different microbial species including commensal strains of Escherichia coli. Bacterial extracellular components of mixed culture can modulate the influence of bacteria on the neutrophil functions. The objective of this study was to compare the effect of pyocyanin, pyoverdine, LPS, exopolysaccharide of single species and mixed culture supernatants of PA strains and E. coli K12 on microbicidal, secretory activity of human neutrophils in vitro. Bacterial components of E. coli K12 in mixed supernatants with 'biofilm' PA strains (PA ATCC, PA BALG) enhanced short-term microbicidal mechanisms and inhibited neutrophil secretion delayed in time. The influence of 'planktonic' PA (PA 9-3) exometabolites in mixed culture is almost mimicked by E. coli K12 effect on functional neutrophil changes. This investigation may help to understand some of the mechanisms of neutrophil response to mixed infections of different PA with other bacteria species. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

The Multi-center Evaluation of the Accuracy of the Contrast MEdium INduced Pd/Pa RaTiO in Predicting FFR (MEMENTO-FFR) Study.

Science.gov (United States)

Leone, Antonio Maria; Martin-Reyes, Roberto; Baptista, Sergio B; Amabile, Nicolas; Raposo, Luis; Franco Pelaez, Juan Antonio; Trani, Carlo; Cialdella, Pio; Basile, Eloisa; Zimbardo, Giuseppe; Burzotta, Francesco; Porto, Italo; Aurigemma, Cristina; Rebuzzi, Antonio G; Faustino, Mariana; Niccoli, Giampaolo; Abreu, Pedro F; Slama, Michel S; Spagnoli, Vincent; Telleria Arrieta, Miren; Amat Santos, Ignacio J; de la Torre Hernandez, Jose M; Lopez Palop, Ramon; Crea, Filippo

2016-08-20

Adenosine administration is needed for the achievement of maximal hyperaemia fractional flow reserve (FFR) assessment. The objective was to test the accuracy of Pd/Pa ratio registered during submaximal hyperaemia induced by non-ionic contrast medium (contrast FFR [cFFR]) in predicting FFR and comparing it to the performance of resting Pd/Pa in a collaborative registry of 926 patients enrolled in 10 hospitals from four European countries (Italy, Spain, France and Portugal). Resting Pd/Pa, cFFR and FFR were measured in 1,026 coronary stenoses functionally evaluated using commercially available pressure wires. cFFR was obtained after intracoronary injection of contrast medium, while FFR was measured after administration of adenosine. Resting Pd/Pa and cFFR were significantly higher than FFR (0.93±0.05 vs. 0.87±0.08 vs. 0.84±0.08, ptime and costs.

Jet evolution in a dense medium: event-by-event fluctuations and multi-particle correlations

Science.gov (United States)

Escobedo, Miguel A.; Iancu, Edmond

2017-11-01

We study the gluon distribution produced via successive medium-induced branchings by an energetic jet propagating through a weakly-coupled quark-gluon plasma. We show that under suitable approximations, the jet evolution is a Markovian stochastic process, which is exactly solvable. For this process, we construct exact analytic solutions for all the n-point correlation functions describing the gluon distribution in the space of energy [M. A. Escobedo, E. Iancu, Event-by-event fluctuations in the medium-induced jet evolution, JHEP 05 (2016) 008. arXiv:arxiv:arXiv:1601.03629, doi:http://dx.doi.org/10.1007/JHEP05(2016)008, M. A. Escobedo, E. Iancu, Multi-particle correlations and KNO scaling in the medium-induced jet evolution, JHEP 12 (2016) 104. arXiv:arxiv:arXiv:1609.06104, doi:http://dx.doi.org/10.1007/JHEP12(2016)104]. Using these results, we study the event-by-event distribution of the energy lost by the jet at large angles and of the multiplicities of the soft particles which carry this energy. We find that the event-by-event fluctuations are huge: the standard deviation in the energy loss is parametrically as large as its mean value [M. A. Escobedo, E. Iancu, Event-by-event fluctuations in the medium-induced jet evolution, JHEP 05 (2016) 008. arXiv:arxiv:arXiv:1601.03629, doi:http://dx.doi.org/10.1007/JHEP05(2016)008]. This has important consequences for the phenomenology of di-jet asymmetry in Pb+Pb collisions at the LHC: it implies that the fluctuations in the branching process can contribute to the measured asymmetry on an equal footing with the geometry of the di-jet event (i.e. as the difference between the in-medium path lengths of the two jets). We compute the higher moments of the multiplicity distribution and identify a remarkable regularity known as Koba-Nielsen-Olesen (KNO) scaling [M. A. Escobedo, E. Iancu, Multi-particle correlations and KNO scaling in the medium-induced jet evolution, JHEP 12 (2016) 104. arXiv:arxiv:arXiv:1609.06104, doi

Jet evolution in a dense medium: event-by-event fluctuations and multi-particle correlations

International Nuclear Information System (INIS)

Escobedo, Miguel A.; Iancu, Edmond

2017-01-01

We study the gluon distribution produced via successive medium-induced branchings by an energetic jet propagating through a weakly-coupled quark-gluon plasma. We show that under suitable approximations, the jet evolution is a Markovian stochastic process, which is exactly solvable. For this process, we construct exact analytic solutions for all the n-point correlation functions describing the gluon distribution in the space of energy [M. A. Escobedo, E. Iancu, Event-by-event fluctuations in the medium-induced jet evolution, JHEP 05 (2016) 008. arXiv: (arXiv:1601.03629), doi: (http://dx.doi.org/10.1007/JHEP05(2016)008), M. A. Escobedo, E. Iancu, Multi-particle correlations and KNO scaling in the medium-induced jet evolution, JHEP 12 (2016) 104. arXiv: (arXiv:1609.06104), doi: (http://dx.doi.org/10.1007/JHEP12(2016)104)]. Using these results, we study the event-by-event distribution of the energy lost by the jet at large angles and of the multiplicities of the soft particles which carry this energy. We find that the event-by-event fluctuations are huge: the standard deviation in the energy loss is parametrically as large as its mean value [M. A. Escobedo, E. Iancu, Event-by-event fluctuations in the medium-induced jet evolution, JHEP 05 (2016) 008. arXiv: (arXiv:1601.03629), doi: (http://dx.doi.org/10.1007/JHEP05(2016)008)]. This has important consequences for the phenomenology of di-jet asymmetry in Pb+Pb collisions at the LHC: it implies that the fluctuations in the branching process can contribute to the measured asymmetry on an equal footing with the geometry of the di-jet event (i.e. as the difference between the in-medium path lengths of the two jets). We compute the higher moments of the multiplicity distribution and identify a remarkable regularity known as Koba-Nielsen-Olesen (KNO) scaling [M. A. Escobedo, E. Iancu, Multi-particle correlations and KNO scaling in the medium-induced jet evolution, JHEP 12 (2016) 104. arXiv: (arXiv:1609.06104), doi: (http

Evaluation of Synergistic Interactions Between Cell-Free Supernatant of Lactobacillus Strains and Amikacin and Genetamicin Against Pseudomonas aeruginosa.

Science.gov (United States)

Aminnezhad, Sargol; Kermanshahi, Rouha Kasra; Ranjbar, Reza

2015-04-01

The indiscriminate use of antibiotics in the treatment of infectious diseases can increase the development of antibiotic resistance. Therefore, there is a big demand for new sources of antimicrobial agents and alternative treatments for reduction of antibiotic dosage required to decrease the associated side effects. In this study, the synergistic action of aminoglycoside antibiotics and cell-free supernatant (CFS) of probiotic (Lactobacillus rahmnosus and L. casei) against Pseudomonas aeruginosa PTCC 1430 was evaluated. A growth medium for culturing of probiotic bacteria was separated by centrifugation. The antimicrobial effects of CFS of probiotic bacteria were evaluated using the agar well diffusion assay. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were evaluated using the micro dilution method. Finally, an interaction between CFS and amikacin or gentamicin against P. aeruginosa PTCC 1430 was examined through the checkerboard method and fractional inhibitory concentration (FIC). Furthermore, CFSs from Lactobacillus strains were analyzed by reversed phase HPLC (RP-HPLC) for antimicrobial compounds. The results showed a significant effect of CFS on the growth of P. aeruginosa. The MIC and MBC of CFS from L. casei were 62.5 µL⁄mL while the MIC and MBC of CFS from L. rhamnosus were 62.5 μL⁄mL and 125 μL⁄mL, respectively. Using the FIC indices, synergistic interactions were observed in combination of CFS and antibiotics. Fractional Inhibitory Concentration indices of CFS from L. casei and aminoglycoside antibiotics were 0.124 and 0.312 while FIC indices of CFS from L. rhamnosus and aminoglycoside antibiotics were 0.124 and 0.56, respectively showing a synergism effect. The results of RP-HPLC showed that CFS of Lactobacillus strains contained acetic acid, lactic acid and hydrogen peroxide (H2O2). Our findings indicate that probiotic bacterial strains of Lactobacillus have a significant inhibitory effect on the

Yoghurt Sinbiotik Berbasis Probiotik Lokal Dapat Mencegah Diare dan Mengubah Status Hematologi Tikus (SYNBIOTIC YOGHURT BASED ON INDIGENOUS PROBIOTIC: IT’S EFFECT ON DIARRHEA AND HEMATOLOGICAL STATUS IN RATS

Directory of Open Access Journals (Sweden)

Made Astawan

2014-08-01

Full Text Available The objective of this study was to observe the effect of functional synbiotics yoghurt (made ofindigenous probiotic + oligofructosaccharides in rats that were infected with Enteropathogenic Escherichiacoli (EPEC, and to detect its effect on hematological status. A total of 25 male Sprague Dawley rats wereused in this study and divided into five treatment groups: (i negative-control; (ii positive-control; (iiisynbiotic yoghurt; (iv synbiotic yoghurt + EPEC; and (v conventional prebiotic yoghurt. Yoghurt (109 cfu/mL lactic acid bacteria was given orally using feeding tube at day one until day 4. At day 8 – day 14animals were infected with EPEC (107 cfu/mL orally using feeding tube; at day 22 blood samples werecollected for hematology analysis (erythrocytes, hematocrit (Hct, hemoglobulin (Hb, platelets, andleucocytes. Post infection with the EPEC diarrhea was developed in rats both in the control positivegroups and in the synbiotic yoghurt + EPEC group. Animals in the positive-control group had a significantlyhigher thrombocytes and leucocytes counts and Hct compared to that in animals in the other groups(P<0.01. Whereas there were no significant effect on the erythrocytes counts and Hb (P>0.05. The treatmentwith synbiotic yoghurt, in both synbiotic yoghurt only and synbiotic yoghurt + EPEC groups significantlyreduced the platelets and leucocytes counts and Hct of rats

Photon mass drag and the momentum of light in a medium

DEFF Research Database (Denmark)

Partanen, Mikko; Häyrynen, Teppo; Oksanen, Jani

2017-01-01

Conventional theories of electromagnetic waves in a medium assume that the energy propagating with the light pulse in the medium is entirely carried by the field. Thus, the possibility that the optical force field of the light pulse would drive forward an atomic mass density wave (MDW...... field. We show that a light pulse having a total electromagnetic energy (h) over bar omega propagating in a nondispersive medium transfers a mass equal to delta m = (n(2) - 1) (h) over bar omega/c(2), where n is the refractive index. MDW, which carries this mass, consists of atoms, which are more......) and the related kinetic and elastic energies is neglected. In this work, we present foundations of a covariant theory of light propagation in a medium by considering a light wave simultaneously with the dynamics of the medium atoms driven by optoelastic forces between the induced dipoles and the electromagnetic...

Out of Medium Fragmentation from Long-Lived Jet Showers

CERN Document Server

Casalderrey-Solana, Jorge; Quiroga-Arias, Paloma

2012-01-01

We study the time structure of vacuum jet evolution via a simple uncertainty principle estimate in the kinematic range explored by current heavy ion collisions at the LHC. We observe that a large fraction of the partonic splittings occur at large times, of the order of several fm. We compare the time distribution of vacuum splittings with the distribution of path lengths traversed by jets in a heavy ion collision. We find that if no medium induced modification of the jet dynamics were present, a very large fraction (larger than 80% for inclusive jets) of the jet splittings would occur outside of the medium. We confront this observation with current available data on jet properties in heavy ion collisions and discuss its implications for the dynamics of jet-medium interactions.

Out of medium fragmentation from long-lived jet showers

Energy Technology Data Exchange (ETDEWEB)

Casalderrey-Solana, Jorge, E-mail: jorge.casalderrey@cern.ch [Departament d' Estructura i Constituents de la Materia, Universitat de Barcelona, Marti i Franques 1, 08028 Barcelona (Spain); Physics Department, Theory Unit, CERN, CH-1211 Geneve 23 (Switzerland); Milhano, Jose Guilherme, E-mail: guilherme.milhano@ist.utl.pt [CENTRA, Instituto Superior Tecnico, Universidade Tecnica de Lisboa, Av. Rovisco Pais, P-1049-001 Lisboa (Portugal); Physics Department, Theory Unit, CERN, CH-1211 Geneve 23 (Switzerland); Quiroga-Arias, Paloma, E-mail: pquiroga@lpthe.jussieu.fr [LPTHE, UPMC Univ. Paris 6 and CNRR UMR7589, Paris (France)

2012-03-29

We study the time structure of vacuum jet evolution via a simple uncertainty principle estimate in the kinematic range explored by current heavy ion collisions at the LHC. We observe that a large fraction of the partonic splittings occur at large times, of the order of several fm. We compare the time distribution of vacuum splittings with the distribution of path lengths traversed by jets in a heavy ion collision. We find that if no medium induced modification of the jet dynamics were present, a very large fraction (larger than 80% for inclusive jets) of the jet splittings would occur outside of the medium. We confront this observation with current available data on jet properties in heavy ion collisions and discuss its implications for the dynamics of jet-medium interactions.

Out of medium fragmentation from long-lived jet showers

International Nuclear Information System (INIS)

Casalderrey-Solana, Jorge; Milhano, José Guilherme; Quiroga-Arias, Paloma

2012-01-01

We study the time structure of vacuum jet evolution via a simple uncertainty principle estimate in the kinematic range explored by current heavy ion collisions at the LHC. We observe that a large fraction of the partonic splittings occur at large times, of the order of several fm. We compare the time distribution of vacuum splittings with the distribution of path lengths traversed by jets in a heavy ion collision. We find that if no medium induced modification of the jet dynamics were present, a very large fraction (larger than 80% for inclusive jets) of the jet splittings would occur outside of the medium. We confront this observation with current available data on jet properties in heavy ion collisions and discuss its implications for the dynamics of jet-medium interactions.

Blackbody Emission from Light Interacting with an Effective Moving Dispersive Medium

OpenAIRE

Petev, M.; Westerberg, N.; Moss, D.; Rubino, E.; Rimoldi, C.; Cacciatori, S. L.; Belgiorno, F.; Faccio, D.

2013-01-01

Intense laser pulses excite a nonlinear polarisation response that may create an effective flowing medium and, under appropriate conditions, a blocking horizon for light. Here we analyse in detail the interaction of light with such laser-induced flowing media, fully accounting for the medium dispersion properties. An analytical model based on a first Born-approximation is found to be in excellent agreement with numerical simulations based on Maxwell's equations and shows that when a blocking ...

Indigenous lactobacilli strains of food and human sources reverse enteropathogenic E. coli O26:H11-induced damage in intestinal epithelial cell lines: effect on redistribution of tight junction proteins.

Science.gov (United States)

Jariwala, Ruchi; Mandal, Hemanti; Bagchi, Tamishraha

2017-09-01

The aim of the study was to investigate the neutralizing effect of lactobacilli isolated from indigenous food and human sources on enteropathogenic Escherichia coli (EPEC) O26 : H11-induced epithelial barrier dysfunction in vitro. This was assessed by transepithelial electrical resistance (TEER) and permeability assays using intestinal cell lines, HT-29 and Caco-2. Furthermore, the expression and distribution of tight junction (TJ) proteins were analysed by qRT-PCR and immunofluorescence assay, respectively. The nine strains used in the study were from different species viz. Lactobacillus fermentum, Lactobacillushelveticus, Lactobacillus salivarius and Lactobacillus plantarum. All strains were able to reverse the decrease in TEER and corresponding increase in permeability across E. coli-infected monolayers. Maximum reversal was observed after 18 h [up to 93.8±2.0 % by L. rhamnosus GG followed by L. fermentum IIs11.2 (92.6±2.2 %) and L. plantarum GRI-2 (91.9±0.9 %)] of lactobacilli exposure following EPEC O26 : H11 infection. All strains were able to redistribute the TJ proteins to the cell periphery either partially or completely. Moreover, L. helveticus FA-7 was also able to significantly increase the mRNA expression of ZO-1 and claudin-1 (2.5-fold and 3.0-fold, respectively; PGRI-2 were good in all the aspects studied, and the other strains were good in some aspects. L. helveticus FA-7, L. fermentum FA-1 and L. plantarum GRI-2 can therefore be used for potential therapeutic purpose against intestinal epithelial dysfunction.

Contrast medium-induced nephropathy: Aspects on incidence ...

African Journals Online (AJOL)

Contrast media-induced nephropathy (CIN) is a well-known complication of radiological examinations employing iodine contrast media (I-CM). The rapid development and frequent use of coronary interventions and multi-channel detector computed tomography with concomitant administration of relatively large doses of ...

1,25-dihydroxyvitamin D3 induces LL-37 and HBD-2 production in keratinocytes from diabetic foot ulcers promoting wound healing: an in vitro model.

Directory of Open Access Journals (Sweden)

Irma Gonzalez-Curiel

Full Text Available Diabetic foot ulcers (DFU are one of the most common diabetes-related cause of hospitalization and often lead to severe infections and poor healing. It has been recently reported that patients with DFU have lower levels of antimicrobial peptides (AMPs at the lesion area, which contributes with the impairment of wound healing. The aim of this study was to determine whether 1,25-dihydroxyvitamin D3 (1,25 (OH2 D3 and L-isoleucine induced HBD-2 and LL-37 in primary cultures from DFU. We developed primary cell cultures from skin biopsies from 15 patients with DFU and 15 from healthy donors. Cultures were treated with 1,25 (OH2D3 or L-isoleucine for 18 h. Keratinocytes phenotype was identified by western blot and flow cytometry. Real time qPCR for DEFB4, CAMP and VDR gene expression was performed as well as an ELISA to measure HBD-2 and LL-37 in supernatant. Antimicrobial activity, in vitro, wound healing and proliferation assays were performed with conditioned supernatant. The results show that primary culture from DFU treated with 1,25(OH2D3, increased DEFB4 and CAMP gene expression and increased the production of HBD-2 and LL-37 in the culture supernatant. These supernatants had antimicrobial activity over E. coli and induced remarkable keratinocyte migration. In conclusion the 1,25(OH2D3 restored the production of AMPs in primary cell from DFU which were capable to improve the in vitro wound healing assays, suggesting their potential therapeutic use on the treatment of DFU.

Co-administration of plasmid expressing IL-12 with 14-kDa Schistosoma mansoni fatty acid-binding protein cDNA alters immune response profiles and fails to enhance protection induced by Sm14 DNA vaccine alone.

Science.gov (United States)

Fonseca, Cristina T; Pacífico, Lucila G G; Barsante, Michele M; Rassi, Tatiana; Cassali, Geovanni D; Oliveira, Sérgio C

2006-08-01

Schistosomiasis is an endemic disease that affects 200 million people worldwide. DNA-based vaccine is a promising strategy to induce protective immunity against schistosomiasis, since both humoral and cellular immune responses are involved in parasite elimination. In this study, we evaluated the ability of Sm14 cDNA alone or in association with a plasmid expressing murine interleukin (IL)-12 to induce protection against challenge infection. Mice were immunized with four doses of the DNA vaccine and the levels of protection were determined by worm burden recovery after challenge infection. Specific antibody production to rSm14 was determined by ELISA, and cytokine production was measured in splenocyte culture supernatants stimulated with rSm14 and in bronchoalveolar lavage of vaccinated mice after challenge infection. DNA immunization with pCI/Sm14 alone induced 40.5% of worm reduction. However, the use of pCI/IL-12 as adjuvant to pCI/Sm14 immunization failed to enhance protection against challenge infection. Protection induced by pCI/Sm14 immunization correlates with specific IgG antibody production against Sm14, Th1 type of immune response with high levels of interferon (IFN)-gamma and low levels of IL-4 in splenocyte culture supernatants and in bronchoalveolar lavage after challenge infection. IL-12 co-administration with pCI/Sm14 induced a significant production of nitric oxide in splenocyte culture supernatants and also lymphocyte suppression, with reduced percentage of T cells producing IFN-gamma and tumor necrosis factor-alpha.

Biosynthesis, characterization and antibacterial activity of silver nanoparticles using an endophytic fungal supernatant of Raphanus sativus

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Tej Singh

2017-06-01

Full Text Available In this study, biological synthesis of silver nanoparticles (AgNPs from supernatant of endophytic fungus Alternaria sp. isolated from the healthy leaves of Raphanus sativus is studied. The synthesized AgNPs are characterized using UV-vis spectroscopy and Fourier transform-infrared spectroscopy (FTIR. The structural analysis is done by powder X-ray diffraction (XRD method. The stability of AgNPs is studied by dynamic light scattering (DLS method. The size and shape of AgNPs are observed by transmission electron microscopy (TEM and atomic force microscopy (AFM and found to be spherical with an average particles size of 4–30 nm. Further, these AgNPs have been found to be highly toxic against human pathogenic bacteria, suggesting the possibility of using AgNPs as efficient antibacterial agents.

Radiation-induced apoptosis in different pH environments in vitro

International Nuclear Information System (INIS)

Lee, Hyung-Sik; Park, Heon J.; Lyons, John C.; Griffin, Robert J.; Auger, Elizabeth A.; Song, Chang W.

1997-01-01

Purpose: The effect of environmental pH on the radiation-induced apoptosis in tumor cells in vitro was investigated. Methods and Materials: Mammary adenocarcinoma cells of A/J mice (SCK cells) were irradiated with γ-rays using a 137 Cs irradiator and incubated in media of different pHs. After incubation at 37 deg. C for 24-120 h the extent of apoptosis was determined using agarose gel electrophoresis, TdT-mediated dUTP-biotin nick end labeling (TUNEL) staining, flow cytometry, and release of 3 H from 3 H-thymidine labeled cells. The clonogenicity of the cells irradiated in different pH medium was determined, and the progression of cells through the cell cycle after irradiation in different pHs was also determined with flow cytometry. Results: Irradiation with 2-12 Gy of γ-rays induced apoptosis in SCK cells in pH 7.5 medium within 48 h as judged from the results of four different assays mentioned. Radiation-induced apoptosis declined as the medium pH was lowered from 7.5 to 6.4. Specifically, the radiation-induced degradation of DNA including the early DNA breaks, as determined with the TUNEL method, progressively declined as the medium pH was lowered so that little DNA fragmentation occurred 48 h after irradiation with 12 Gy in pH 6.6 medium. When the cells were irradiated and incubated for 48 h in pH 6.6 medium and the medium was replaced with pH 7.5 medium, DNA fragmentation promptly occurred. DNA fragmentation also occurred even in pH 6.6 medium when the cells were irradiated and maintained in pH 7.5 medium for 8 h or longer post-irradiation before incubation in pH 6.6 medium. The radiation-induced G 2 arrest in pH 6.6 medium lasted markedly longer than that in pH 7.5 medium. Conclusion: Radiation-induced apoptosis in SCK cells in vitro is reversibly suppressed in an acidic environment. Taking the results of four different assays together, it was concluded that early step(s) in the apoptotic pathway, probably the DNA break or upstream of DNA break, is

Contrast induced nephropathy

DEFF Research Database (Denmark)

Stacul, Fulvio; van der Molen, Aart J; Reimer, Peter

2011-01-01

PURPOSE: The Contrast Media Safety Committee (CMSC) of the European Society of Urogenital Radiology (ESUR) has updated its 1999 guidelines on contrast medium-induced nephropathy (CIN). AREAS COVERED: Topics reviewed include the definition of CIN, the choice of contrast medium, the prophylactic me...

Changes in the ammonia-oxidizing bacteria community in response to operational parameters during the treatment of anaerobic sludge digester supernatant.

Science.gov (United States)

Cydzik-Kwiatkowska, Agnieszka; Zielińska, Magdalena; Bernat, Katarzyna; Kulikowska, Dorota; Wojnowska-Baryła, Irena

2012-07-01

The understanding of the relationship between ammoniaoxidizing bacteria (AOB) communities in activated sludge and the operational treatment parameters supports the control of the treatment of ammonia-rich wastewater. The modifications of treatment parameters by alteration of the number and length of aerobic and anaerobic stages in the sequencing batch reactor (SBR) working cycle may influence the efficiency of ammonium oxidation and induce changes in the AOB community. Therefore, in the research, the impact of an SBR cycle mode with alternating aeration/ mixing conditions (7 h/1 h vs. 4 h/5.5 h) and volumetric exchange rate (n) on AOB abundance and diversity in activated sludge during the treatment of anaerobic sludge digester supernatant at limited oxygen concentration in the aeration stage (0.7 mg O2/l) was assessed. AOB diversity expressed by the Shannon-Wiener index (H') was determined by the cycle mode. At aeration/mixing stage lengths of 7 h/1 h, H' averaged 2.48 +/- 0.17, while at 4 h/ 5.5 h it was 2.35 +/- 0.16. At the given mode, AOB diversity decreased with increasing n. The cycle mode did not affect AOB abundance; however, a higher AOB abundance in activated sludge was promoted by decreasing the volumetric exchange rate. The sequences clustering with Nitrosospira sp. NpAV revealed the uniqueness of the AOB community and the simultaneously lower ability of adaptation of Nitrosospira sp. to the operational parameters applied in comparison with Nitrosomonas sp.

Effect of medium replenishment or composition on [3H] thymidine incorporation in uv-irradiated CHO-K1 cells

International Nuclear Information System (INIS)

Newman, C.N.; Miller, J.H.

1985-03-01

Because culture medium contains uv-absorbing material, it is usually removed just before uv-irradiation of tissue culture monolayers. However, medium removal and replenishment with fresh medium alone (sham-irradiation) causes up to a 10-fold reduction in the rate of [ 3 H]TdR incorporation in CHO-K1 cells which persists for several hours. This reduction, which is much smaller ( 3 H]TdR pulse-label in conditioned (spent) and in fresh medium; TdR in the former is converted by cells to thymine. When responses of uv-irradiated cells are normalized to responses of corresponding sham-irradiated cultures, considerable variation is observed in replicate experiments because fresh medium appears to induce transient metabolic imbalances in irradiated cells which are not readily controlled. This problem can, in part, be circumvented by replenishing treated cultures with the original spent medium; however, the presence of CdR in the growth medium still causes an anomalous 2-3-fold greater uv-induced reduction in [ 3 H]TdR incorporation than is observed in the absence of CdR. 17 refs., 3 figs., 1 tab

Contrast induced nephropathy

DEFF Research Database (Denmark)

Stacul, Fulvio; van der Molen, Aart J; Reimer, Peter

2011-01-01

PURPOSE: The Contrast Media Safety Committee (CMSC) of the European Society of Urogenital Radiology (ESUR) has updated its 1999 guidelines on contrast medium-induced nephropathy (CIN). AREAS COVERED: Topics reviewed include the definition of CIN, the choice of contrast medium, the prophylactic me....../min/1.73 m (2) is CIN risk threshold for intravenous contrast medium. • Hydration with either saline or sodium bicarbonate reduces CIN incidence. • Patients with eGFR = 60 ml/min/1.73 m (2) receiving contrast medium can continue metformin normally....

Enhancement of alpha particles-induced cell transformation by oxygen free radicals and tumor necrosis factor released from phagocytes

International Nuclear Information System (INIS)

Gong Yifen; Guo Renfeng; Zhu Maoxiang; Shou Jiang; Ge Guixiu; Yang Zhihua; Hieber, L.; Peters, K.; Schippel, C.

1997-01-01

To illustrate the role of several endogenous factors released from phagocytes under chronic inflammation in radiation-induced cancer. C 3 T 10 T 1/2 and SHE cells were used as targets, and 238 Pu alpha source was used in alpha irradiation. The enhancement of TF in alpha particles-induced cell transformation by PMA-stimulated human blood and zymosan-stimulated U-937 cells was studied using formation of transformed foci. Transformation frequency (TF) of C 3 H 10 T 1/2 cells exposed to alpha particles of 0.5 Gy increased 2.1 and 2.8 fold by PMA-and PMA-stimulated neutrophils, respectively. TF of irradiated SHE cells at a dose of 0.5 Gy increased 12 fold by the addition of the supernatant of macrophage-like U-937 cell line. It was shown that TF of irradiated SHE cells at above dose increased 8 fold by the supernatant treated with anti-TNF-α could be subcultured continuously in vitro. The cells at 40 th passage and two lines of monoclone cells have the ability to develop malignant tumors in nude mice. The overdose of free radicals and TNF-α released from neutrophils and macrophages have played an important role in low dose radiation-induced cancer

Effects of Saccharomyces cerevisiae or boulardii yeasts on acute stress induced intestinal dysmotility.

Science.gov (United States)

West, Christine; Stanisz, Andrew M; Wong, Annette; Kunze, Wolfgang A

2016-12-28

To investigate the capacity of Saccharomyces cerevisiae ( S. cerevisiae ) and Saccharomyces boulardii ( S. boulardii ) yeasts to reverse or to treat acute stress-related intestinal dysmotility. Adult Swiss Webster mice were stressed for 1 h in a wire-mesh restraint to induce symptoms of intestinal dysmotility and were subsequently killed by cervical dislocation. Jejunal and colon tissue were excised and placed within a tissue perfusion bath in which S. cerevisiae , S. boulardii , or their supernatants were administered into the lumen. Video recordings of contractility and gut diameter changes were converted to spatiotemporal maps and the velocity, frequency, and amplitude of propagating contractile clusters (PCC) were measured. Motility pre- and post-treatment was compared between stressed animals and unstressed controls. S. boulardii and S. cerevisiae helped to mediate the effects of stress on the small and large intestine. Restraint stress reduced jejunal transit velocity (mm/s) from 2.635 ± 0.316 to 1.644 ± 0.238, P boulardii helped to restore jejunal and colonic velocity towards the unstressed controls; 1.833 ± 0.688 to 2.627 ± 0.664, P boulardii or S. cerevisiae supernatants also helped to restore motility to unstressed values in similar capacity. There is a potential therapeutic role for S. cerevisiae and S. boulardii yeasts and their supernatants in the treatment of acute stress-related gut dysmotility.

Distribution of espM and espT among enteropathogenic and enterohaemorrhagic Escherichia coli

Science.gov (United States)

Arbeloa, Ana; Blanco, Miguel; Moreira, Fabiana C.; Bulgin, Richard; López, Cecilia; Dahbi, Ghizlane; Blanco, Jesús E.; Mora, Azucena; Alonso, María Pilar; Mamani, Rosalia Ceferina; Gomes, Tânia A. T.; Blanco, Jorge; Frankel, Gad

2009-01-01

Enterohaemorrhagic Escherichia coli (EHEC) and enteropathogenic E. coli (EPEC) translocate dozens of type III secretion system effectors, including the WxxxE effectors Map, EspM and EspT that activate Rho GTPases. While map, which is carried on the LEE pathogenicity island, is absolutely conserved among EPEC and EHEC strains, the prevalence of espM and espT is not known. Here we report the results of a large screen aimed at determining the prevalence of espM and espT among clinical EPEC and EHEC isolates. The results suggest that espM, detected in 51 % of the tested strains, is more commonly found in EPEC and EHEC serogroups that are linked to severe human infections. In contrast, espT was absent from all the EHEC isolates and was found in only 1.8 % of the tested EPEC strains. Further characterization of the virulence gene repertoire of the espT-positive strains led to the identification of a new ζ2 intimin variant. All the espT-positive strains but two contained the tccP gene. espT was first found in Citrobacter rodentium and later in silico in EPEC E110019, which is of particular interest as this strain was responsible for a particularly severe diarrhoeal outbreak in Finland in 1987 that affected 650 individuals in a school complex and an additional 137 associated household members. Comparing the protein sequences of EspT to that of E110019 showed a high level of conservation, with only three strains encoding EspT that differed in 6 amino acids. At present, it is not clear why espT is so rare, and what impact EspM and EspT have on EPEC and EHEC infection. PMID:19528152

Characterization of a human antigen specific helper factor

International Nuclear Information System (INIS)

Richardson, B.

1986-01-01

While antigen (Ag) specific helper factors have been characterized in mice, similar molecules have not been identified in humans. To characterize human antigen specific helper molecules, an IL-2 dependent tetanus toxoid (T.T.) reactive T cell line was fused with a 6-thioguanine resistant CEM line, and hybrids selected in medium containing hypoxanthine and azaserine. Hybrids were screened by culturing the cells with 35 S-Met then reacting the supernatants with T.T. or hepatitis vaccine immobilized on nitrocellulose. One hybrid, TT6BA-O, was identified which secreted a Met-containing molecule which bound T.T. but not hepatitis vaccine. Supernatants from TT6BA-O, but not the parent CEM line, when added to autologous peripheral blood mononuclear cells (PBMC's) stimulated secretion of T.T. specific antibodies (Abs). Specificity controls demonstrated that TT6BA-O supernatant did not induce antibodies to diphtheria toxoid, hepatitis vaccine or pneumococcal polysaccharide, and total immunoglobulin (lg) synthesis was minimally increased. In contrast, pokeweed mitogen stimulated significant lg synthesis as well as Ab's to pneumococcal polysaccharide and T.T. TT6BA-O supernatant induced anti-T.T.Ab's in autologous PBMC's but not PBMC's from 3 unrelated donors, suggesting that the activity of the helper factor is restricted, possibly by the MHC. The molecular weight of the helper factor was estimated at 100,000-150,000 by Sephacryl S-300 chromatography. Finally, the helper factor could be demonstrated to bind and elute from sephorose-immobilized T.T. and anti-DR antisera, but not anti-lg antisera or the T40/25 monoclonal antibody, which binds a nonpolymorphic determinant on the human T cell receptor. These results demonstrate that human Ag specific helper factors exist, bind antigen and bear class II MHC determinants

Analysis of glutathione in supernatants and lysates of a human proximal tubular cell line from perfusion culture upon intoxication with cadmium chloride by HPLC and LC-ESI-MS

NARCIS (Netherlands)

Hahn, Hans; Huck, Christian W; Rainer, Matthias; Najam-ul-Haq, Muhammad; Bakry, Rania; Abberger, Thomas; Jennings, Paul; Pfaller, Walter; Bonn, Günther K

A simple and highly effective reversed-phase (RP) high-performance liquid chromatography (HPLC) method is described for analysing glutathione (GSH) and glutathione disulfide (GSSG) in out-flowing supernatants and lysates of perfusion cell cultures of human kidney cells (HK-2 cells) continuously

Filarial excretory-secretory products induce human monocytes to produce lymphangiogenic mediators.

Directory of Open Access Journals (Sweden)

Tiffany Weinkopff

2014-07-01

Full Text Available The nematodes Wuchereria bancrofti and Brugia spp. infect over 120 million people worldwide, causing lymphedema, elephantiasis and hydrocele, collectively known as lymphatic filariasis. Most infected individuals appear to be asymptomatic, but many exhibit sub-clinical manifestations including the lymphangiectasia that likely contributes to the development of lymphedema and elephantiasis. As adult worm excretory-secretory products (ES do not directly activate lymphatic endothelial cells (LEC, we investigated the role of monocyte/macrophage-derived soluble factors in the development of filarial lymphatic pathology. We analyzed the production of IL-8, IL-6 and VEGF-A by peripheral blood mononuclear cells (PBMC from naïve donors following stimulation with filarial ES products. ES-stimulated PBMCs produced significantly more IL-8, IL-6 and VEGF-A compared to cells cultured in medium alone; CD14(+ monocytes appear to be the primary producers of IL-8 and VEGF-A, but not IL-6. Furthermore, IL-8, IL-6 and VEGF-A induced in vitro tubule formation in LEC Matrigel cultures. Matrigel plugs supplemented with IL-8, IL-6, VEGF-A, or with supernatants from ES-stimulated PBMCs and implanted in vivo stimulated lymphangiogenesis. Collectively, these data support the hypothesis that monocytes/macrophages exposed to filarial ES products may modulate lymphatic function through the secretion of soluble factors that stimulate the vessel growth associated with the pathogenesis of filarial disease.

Kinetics of phototoxicity of Fischer's medium for L5178Y leukemic cells

International Nuclear Information System (INIS)

Griffin, F.M.; Ashland, G.; Capizzi, R.L.

1981-01-01

The uncontrolled exposure of Fischer's medium to cool white fluorescent (CWF) light or other sources emitting near-ultraviolet or visible light absorbance by riboflavin is a crucial random variable in experiments which utilize L5178Y cells and this medium. The radiation effects of CWF light result in the rapid development of toxic photoproducts in the medium which are cytostatic at lower doses of radiation and cytotoxic at higher doses. After a 24-hr suspension in medium irradiated for 3 or 48 hr, the cloning efficiencies of cells subsequently plated in light-protected medium were 87 and 3%, respectively. The corresponding near-ultraviolet doses for these periods of exposure to CWF light were 0.22 x 10(4) for a 3-hr exposure and 3.47 x 10(4) J/sq m for a 48-hr exposure. Cells incubated in lightly irradiated medium resumed growth at nearly normal rates following a 24- to 48-hr period in which no increase in cell numbers occurred. Exposure of medium containing riboflavin, but not tryptophan or tyrosine, to CWF light also produces toxic medium. Tryptophan enhances riboflavin-induced phototoxicity, whereas tyrosine diminishes this effect. As photosusceptibility of this system is very high, Fischer's medium must be fully protected from all sources of light absorbable by riboflavin

Chaotic wave trains in an oscillatory/excitable medium

International Nuclear Information System (INIS)

Rabinovitch, A.; Gutman, M.; Biton, Y.; Aviram, I.

2006-01-01

We study the chaotic dynamics of a heterogeneous reaction-diffusion medium composed of two uniform regions: one oscillatory, and the other excitable. It is shown that, by altering the diffusion coefficient, local chaotic oscillations can be induced at the interface between regions, which in turn, generate different chaotic sequences of pulses traveling in the excitable region. We analyze the properties of the local chaotic driver, as well as the diffusion-induced transitions. A procedure based on the abnormal frequency-locking phenomenon is proposed for controlling such sequences. Relevance of the obtained results to cardiac dynamics is briefly discussed

Exosomes Derived from Dendritic Cells Treated with Schistosoma japonicum Soluble Egg Antigen Attenuate DSS-Induced Colitis

Directory of Open Access Journals (Sweden)

Lifu Wang

2017-09-01

Full Text Available Exosomes are 30–150 nm small membrane vesicles that are released into the extracellular medium via cells that function as a mode of intercellular communication. Dendritic cell (DC-derived exosomes modulate immune responses and prevent the development of autoimmune diseases. Moreover, Schistosoma japonicum eggs show modulatory effects in a mouse model of colitis. Therefore, we hypothesized that exosomes derived from DCs treated with S. japonicum soluble eggs antigen (SEA; SEA-treated DC exosomes would be useful for treating inflammatory bowel disease (IBD. Exosomes were purified from the supernatant of DCs treated or untreated with SEA and identified via transmission electron microscopy, western blotting and NanoSight. Acute colitis was induced via the administration of dextran sulfate sodium (DSS in drinking water (5.0%, wt/vol. Treatment with exosomes was conducted via intraperitoneal injection (i.p.; 50 μg per mouse from day 0 to day 6. Clinical scores were calculated based on weight loss, stool type, and bleeding. Colon length was measured as an indirect marker of inflammation, and colon macroscopic characteristics were determined. Body weight loss and the disease activity index of DSS-induced colitis mice decreased significantly following treatment with SEA-treated DC exosomes. Moreover, the colon lengths of SEA-treated DC exosomes treated colitis mice improved, and their mean colon macroscopic scores decreased. In addition, histologic examinations and histological scores showed that SEA-treated DC exosomes prevented colon damage in acute DSS-induced colitis mice. These results indicate that SEA-treated DC exosomes attenuate the severity of acute DSS-induced colitis mice more effectively than DC exosomes. The current work suggests that SEA-treated DC exosomes may be useful as a new approach to treat IBD.

Hemolytic activity of Fusobacterium necrophorum culture supernatants due to presence of phospholipase A and lysophospholipase.

Science.gov (United States)

Abe, P M; Kendall, C J; Stauffer, L R; Holland, J W

1979-01-01

Culture supernatants of Fusobacterium necrophorum demonstrated hemolytic activity. The hemolysin(s), which was partially purified by ammonium sulfate precipitation, was temperature-dependent and heat labile. The spectrum of hemolytic activity against various erythrocytes included rabbit, human, and dog erythrocytes. Goats, sheep, and bovine erythrocytes showed only trace hemolysis. According to results of thin-layer chromatography, the hemolysin hydrolyzed rabbit erythrocyte phosphatidyl choline, phosphatidyl ethanolamine, lysophosphatidyl choline, and bovine phosphatidyl choline. Hydrolysis of egg yolk phosphatidyl choline, bovine phosphatidyl ethanolamine, cholesterol, 1,2-dipalmitin, 1,3-dipalmitin, sphingomyelin, or triolein was not detected by thin layer chromatography. A more sensitive procedure utilizing gas-liquid chromatography revealed that, of the substrates tested, the following were bein hydrolyzed: bovine and egg yolk phosphatidyl choline, lysophosphatidyl choline, alpha-palmito-beta-eleoyl-L-alpha lecithin and alpha-oleoyl-betal-palmitoyl-L-alpha lecithin. Substrates which were weakly hydrolyzed were bovine phosphatidyl ethanolamine, DL-alpha-hosphatidyl ethanolamine dipalmitoyl, 1,2-dipalmitin, 1,3-dipalmitin, and triolein.

Ionizing radiation-induced bystander mutagenesis and adaptation: Quantitative and temporal aspects

International Nuclear Information System (INIS)

Zhang Ying; Zhou Junqing; Baldwin, Joseph; Held, Kathryn D.; Prise, Kevin M.; Redmond, Robert W.; Liber, Howard L.

2009-01-01

This work explores several quantitative aspects of radiation-induced bystander mutagenesis in WTK1 human lymphoblast cells. Gamma-irradiation of cells was used to generate conditioned medium containing bystander signals, and that medium was transferred onto naive recipient cells. Kinetic studies revealed that it required up to 1 h to generate sufficient signal to induce the maximal level of mutations at the thymidine kinase locus in the bystander cells receiving the conditioned medium. Furthermore, it required at least 1 h of exposure to the signal in the bystander cells to induce mutations. Bystander signal was fairly stable in the medium, requiring 12-24 h to diminish. Medium that contained bystander signal was rendered ineffective by a 4-fold dilution; in contrast a greater than 20-fold decrease in the cell number irradiated to generate a bystander signal was needed to eliminate bystander-induced mutagenesis. This suggested some sort of feedback inhibition by bystander signal that prevented the signaling cells from releasing more signal. Finally, an ionizing radiation-induced adaptive response was shown to be effective in reducing bystander mutagenesis; in addition, low levels of exposure to bystander signal in the transferred medium induced adaptation that was effective in reducing mutations induced by subsequent γ-ray exposures.

A noncognate interaction with anti-receptor antibody-activated helper T cells induces small resting murine B cells to proliferate and to secrete antibody

DEFF Research Database (Denmark)

Owens, T

1988-01-01

on resting B cells (even in the presence of intact F23.1 antibody), but could induce antibody secretion by anti-Ig-preactivated B cells. Both F23.1+ clones (E9.D4 and 4.35F2) and one F23.1- clone (D2.2) could synergize with supernatants from activated E9.D4 T cells to induce B cell activation. F(ab')2......Culture of small resting allogeneic B cells (of an irrelevant haplotype) with two clones of T helper (Th) cells that were activated by the F23.1 anti-T cell receptor antibody led to the activation of B cells to proliferate and to secrete antibody. Th cell supernatants by themselves had no effect...... fragments of F23.1 induced E9.D4 to activate B cells as efficiently as intact F23.1 and B cell populations that had been incubated with F23.1 were not activated when cultured with E9.D4, although T cells recognized cell-presented F23.1 and were weakly activated. Reduction of the density of F23.1 adsorbed...

Potential of Pseudoshikonin I Isolated from Lithospermi Radix as Inhibitors of MMPs in IL-1β-Induced SW1353 Cells

Directory of Open Access Journals (Sweden)

Dae Young Lee

2016-08-01

Full Text Available Pseudoshikonin I, the new bioactive constituent of Lithospermi radix, was isolated from this methanol extract by employing reverse-phase medium-pressure liquid chromatography (MPLC using acetonitrile/water solvent system as eluents. The chemical structure was determined based on spectroscopic techniques, including 1D NMR (1H, 13C, DEPT, 2D NMR (gCOSY, gHMBC, gHMQC, and QTOF/MS data. In this study, we demonstrated the effect of pseudoshikonin I on matrix-metalloproteinase (MMPs activation and expression in interleukin (IL-1β-induced SW1353 chondrosarcoma cells. MMPs are considered important for the maintenance of the extracellular matrix. Following treatment with PS, active MMP-1, -2, -3, -9, -13 and TIMP-2 were quantified in the SW1353 cell culture supernatants using a commercially available ELISA kit. The mRNA expression of MMPs in SW1353 cells was measured by RT-PCR. Pseudoshikonin I treatment effectively protected the activation on all tested MMPs in a dose-dependent manner. TIMP-2 mRNA expression was significantly upregulated by pseudoshikonin I treatment. Overall, we elucidated the inhibitory effect of pseudoshikonin on MMPs, and we suggest its use as a potential novel anti-osteoarthritis agent.

Antimicrobial Activity of Cell Free Supernatant of Irradiated Lactic Acid Bacteria Isolates

International Nuclear Information System (INIS)

Abdelaleem, M.A.; AL-Hagar, O.E.Aa.

2015-01-01

Attempts were made to isolate bio preservatives using food wastes with no value and low cost. Whey is the raw material achieved that value. Whey and many other food wastes are used in our study to isolate Lactic acid bacteria (LAB). Cell free supernatants (CFS) of isolates are used to evaluate their antimicrobial activity against indicator pathogenic bacterial strains. CFS-9 isolate from whey has the highest inhibitory activity compared to all other isolates. The inhibitory activity of CFS-9, Nisin (400 IU / ml) and the standard Lactococcus Lactis Subsp. Lactis ATCC 11454 (Lacto) were determined. Furthermore, isolate-9 and Lacto strains were exposed to irradiation at different doses. The inhibition zones of; control isolate-9 (non-irradiated) showed the highest values against all indicator strains, CFS of irradiated Lacto at dose 250 Gy was the highest value against Bacillus cereus and Escherichia coli compared to other irradiation treatments, CFS of irradiated Lacto at dose 100 Gy was the highest value against Staph aureus, while the inhibition zone was in the highest value in CFS of irradiated Lacto at dose 500 Gy against Salmonella typhimurium. Nisin (400 IU / ml) was significantly higher than all CFS of irradiated isolate-9 while, the inhibition zones of all CFS-Lacto (irradiated and nonirradiated) are better and higher than nisin-400

Mechanisms and Consequences of Dopamine Depletion-Induced Attenuation of the Spinophilin/Neurofilament Medium Interaction

Directory of Open Access Journals (Sweden)

Andrew C. Hiday

2017-01-01

Full Text Available Signaling changes that occur in the striatum following the loss of dopamine neurons in the Parkinson disease (PD are poorly understood. While increases in the activity of kinases and decreases in the activity of phosphatases have been observed, the specific consequences of these changes are less well understood. Phosphatases, such as protein phosphatase 1 (PP1, are highly promiscuous and obtain substrate selectivity via targeting proteins. Spinophilin is the major PP1-targeting protein enriched in the postsynaptic density of striatal dendritic spines. Spinophilin association with PP1 is increased concurrent with decreases in PP1 activity in an animal model of PD. Using proteomic-based approaches, we observed dopamine depletion-induced decreases in spinophilin binding to multiple protein classes in the striatum. Specifically, there was a decrease in the association of spinophilin with neurofilament medium (NF-M in dopamine-depleted striatum. Using a heterologous cell line, we determined that spinophilin binding to NF-M required overexpression of the catalytic subunit of protein kinase A and was decreased by cyclin-dependent protein kinase 5. Functionally, we demonstrate that spinophilin can decrease NF-M phosphorylation. Our data determine mechanisms that regulate, and putative consequences of, pathological changes in the association of spinophilin with NF-M that are observed in animal models of PD.

Laser ablation of liquid surface in air induced by laser irradiation through liquid medium

Science.gov (United States)

Utsunomiya, Yuji; Kajiwara, Takashi; Nishiyama, Takashi; Nagayama, Kunihito; Kubota, Shiro; Nakahara, Motonao

2010-10-01

The pulse laser ablation of a liquid surface in air when induced by laser irradiation through a liquid medium has been experimentally investigated. A supersonic liquid jet is observed at the liquid-air interface. The liquid surface layer is driven by a plasma plume that is produced by laser ablation at the layer, resulting in a liquid jet. This phenomenon occurs only when an Nd:YAG laser pulse (wavelength: 1064 nm) is focused from the liquid onto air at a low fluence of 20 J/cm2. In this case, as Fresnel’s law shows, the incident and reflected electric fields near the liquid surface layer are superposed constructively. In contrast, when the incident laser is focused from air onto the liquid, a liquid jet is produced only at an extremely high fluence, several times larger than that in the former case. The similarities and differences in the liquid jets and atomization processes are studied for several liquid samples, including water, ethanol, and vacuum oil. The laser ablation of the liquid surface is found to depend on the incident laser energy and laser fluence. A pulse laser light source and high-resolution film are required to observe the detailed structure of a liquid jet.

Inhibition of the early stage of Salmonella enterica serovar Enteritidis biofilm development on stainless steel by cell-free supernatant of a Hafnia alvei culture.

Science.gov (United States)

Chorianopoulos, Nikos G; Giaouris, Efstathios D; Kourkoutas, Yiannis; Nychas, George-John E

2010-03-01

Compounds present in Hafnia alvei cell-free culture supernatant cumulatively negatively influence the early stage of biofilm development by Salmonella enterica serovar Enteritidis on stainless steel while they also reduce the overall metabolic activity of S. Enteritidis planktonic cells. Although acylhomoserine lactones (AHLs) were detected among these compounds, the use of several synthetic AHLs was not able to affect the initial stage of biofilm formation by this pathogen.

Effects of acetylcysteine and probucol on contrast medium-induced depression of intrinsic renal glutathione peroxidase activity in diabetic rats.

Science.gov (United States)

Yen, Hsueh-Wei; Lee, Hsiang-Chun; Lai, Wen-Te; Sheu, Sheng-Hsiung

2007-04-01

Antioxidants such as N-acetylcysteine and probucol have been used to protect patients from contrast media-induced nephrotoxicity. The mechanisms underlying these protective effects are not well understood. We hypothesized that acetylcysteine and probucol alter the activity of endogenous antioxidant enzyme activity. Four weeks after induction of diabetes with streptozotocin, diabetic and nondiabetic rats were divided into three groups. Group 1 rats did not receive any antioxidant agents. Group 2 rats were treated with acetylcysteine and group 3 rats with probucol for 1 week before injection of the contrast medium diatrizoate (DTZ). We found that diabetic rats had higher renal glutathione peroxidase (GPx) activity than normal rats. DTZ suppressed renal GPx activity significantly in both group 1 diabetic and normal rats. Interestingly, renal GPx activity in both diabetic and normal rats pretreated with acetylcysteine or probucol was not inhibited by DTZ. Renal superoxide dismutase (SOD) increased significantly in normal rats after DTZ injection, but not in diabetic rats. Finally, acetylcysteine or probucol did not significantly influence renal SOD. These findings suggest that the renal protective effects of acetylcysteine and probucol against contrast-induced oxidative stress and nephrotoxicity may be mediated by altering endogenous GPx activity.

Medium-Energy Nuclear Data Library (MENDLIB): Phase 1

International Nuclear Information System (INIS)

Siciliano, E.R.; Arthur, E.D.

1987-10-01

This document describes an initial step towards the formation of a computerized on-line data library, which would contain published medium-energy experimental data, and which would serve the basic and applied needs of the medium-energy nuclear physics community. The data emphasized in this project will be from measured charged-particle and meson induced nuclear scattering and reactions; an area for which no such data base presently exists. Access to the data will be through a menu-driven program in a user-friendly environment. The project is divided into three phases: Phase 1 involves compilation of Clinton P. Anderson Meson Physics Facility (LAMPF) data from nucleon and pion induced reactions, Phase 2 includes nucleon and pion data from other medium-energy facilities, and Phase 3 includes electron, light-ion, and possibly kaon and anti-nucleon data. The initial goals, the manner in which they would be pursued, and the resources needed to implement Phase 1 (the pilot phase) are discussed in detail. Possible expansion of Phase 1 to attain the envisioned goals of Phase 2 and 3 are briefly outlined. During all stages of the project, input from the community will be sought via the various facility user groups and the American Physical Society Division of Nuclear Physics. It is proposed that the Applied Nuclear Science Group (T-2) of the Los Alamos National Laboratory oversees the development and implementation of this project, and the LAMPF VAX computers be used as the host computers for on-line access

Quantitative Real-time Polymerase Chain Reaction for Enteropathogenic Escherichia coli: A Tool for Investigation of Asymptomatic Versus Symptomatic Infections

Science.gov (United States)

Barletta, Francesca; Mercado, Erik; Ruiz, Joaquim; Ecker, Lucie; Lopez, Giovanni; Mispireta, Monica; Gil, Ana I.; Lanata, Claudio F.; Cleary, Thomas G.

2011-01-01

Background. Enteropathogenic Escherichia coli (EPEC) strains are pediatric pathogens commonly isolated from both healthy and sick children with diarrhea in areas of endemicity. The aim of this study was to compare the bacterial load of EPEC isolated from stool samples from children with and without diarrhea to determine whether bacterial load might be a useful tool for further study of this phenomenon. Methods. EPEC was detected by polymerase chain reaction (PCR) of colonies isolated on MacConkey plates from 53 diarrheal and 90 healthy children aged <2 years. DNA was isolated from stool samples by cetyltrimethylammonium bromide extraction. To standardize quantification by quantitative real-time PCR (qRT-PCR), the correlation between fluorescence threshold cycle and copy number of the intimin gene of EPEC E2348/69 was determined. Results. The detection limit of qRT-PCR was 5 bacteria/mg stool. The geometric mean load in diarrhea was 299 bacteria/mg (95% confidence interval [CI], 77–1164 bacteria/mg), compared with 29 bacteria/mg (95% CI, 10–87 bacteria/mg) in control subjects (P = .016). Bacterial load was significantly higher in children with diarrhea than in control subjects among children <12 months of age (178 vs 5 bacteria/mg; P = .006) and among children with EPEC as the sole pathogen (463 vs 24 bacteria/mg; P = .006). Conclusions. EPEC load measured by qRT-PCR is higher in diarrheal than in healthy children. qRT-PCR may be useful to study the relationship between disease and colonization in settings of endemicity. PMID:22028433

Whey permeate fermented with kefir grains shows antifungal effect against Fusarium graminearum.

Science.gov (United States)

Gamba, Raúl Ricardo; De Antoni, Graciela; Peláez, Angela León

2016-05-01

The objective of the work reported here was to study the antifungal capability of cell-free supernatants obtained from whey permeates after fermentation by the kefir grains CIDCA AGK1 against Fusarium graminearum growth and zearalenone (ZEA) production. The assays were performed in order to study the conidial germination inhibition -in liquid media- and the effect on fungal growth rate and the Latency phase -in solid media. We observed that fermented supernatants of pH 3·5 produced the highest percentages of inhibition of conidial germination. The dilution and, particularly, alkalinisation of them led to the gradual loss of antifungal activity. In the fungal inhibition assays on plates we found that only the highest proportion of supernatant within solid medium had significant antifungal activity, which was determined as fungicidal. There was no ZEA biosynthesis in the medium with the highest proportion of supernatant, whereas at lower concentrations, the mycotoxin production was strain-dependent. From the results obtained we concluded that kefir supernatants had antifungal activity on the F. graminearum strains investigated and inhibited mycotoxin production as well, but in a strain-dependent fashion. The present work constitutes the first report of the effect of the products obtained from the kefir-grain fermentation of whey permeates - a readily available by-product of the dairy industry - on F. graminearum germination, growth, and toxin production.

Onset of color decoherence for soft gluon radiation in a medium

Science.gov (United States)

Mehtar-Tani, Y.; Salgado, C. A.; Tywoniuk, K.

2011-12-01

We report on recent studies of the phenomenon of color decoherence in jets in QCD media. The effect is most clearly observed in the radiation pattern of a quark-antiquark antenna, created in the same quantum state, traversing a dense color deconfined plasma. Multiple scattering with the medium color charges gradually destroys the coherence of the antenna. In the limit of opaque media, this ultimately leads to independent radiation off the antenna constituents. Accordingly, radiation off the total charge vanishes implying a memory loss effect induced by the medium.

Recommendation to use iso-osmotic contrast medium in interventional treatment

International Nuclear Information System (INIS)

Zhou Bing; Cheng Yongde

2012-01-01

With the rapid development of imaging diagnostic and interventional therapeutic techniques, the contrast medium (CM) has been used more and more common in clinical practice, and meanwhile more and more attention has been paid to the CM-related adverse events. Contrast induced nephropathy (CN) is the most common CM-related adverse event, and CM-related neurotoxicity has already attracted the physicians' attention. The osmotic pressure of the iso-osmotic contrast medium (IOCM) is quite the same as that of the plasma, and therefore its safety is higher than that of low-osmotic contrast medium (LOCM), the patient's tolerance to IOCM is better than that to LOCM. For this reason, the use of IOCM should be strongly recommended in interventional procedures, which is of great significance to the reduction of the occurrence of CM-related adverse events. (authors)

Impact induced splash and spill in a quasi-confided granular medium

Science.gov (United States)

Ogale, S. B.

2005-03-01

Dissipation of the energy of impact in a granular medium and its effects has been a subject of considerable scientific for quite some time. In this work we have explored and analyzed the splash and spill effects caused by the impact of a ball dropped from a height into a granular medium in a open container. Three different granular media, namely rice, mustard seeds, and cream of wheat were used. The amount of spilled-over granular matter was measured as a function of the ball-drop height. Digital pictures of the splash process were also recorded. The quantity of spilled granular matter varies linearly with the impact energy. However additional step like structures are also noted. Specifically, a distinct and large jump is seen in the spilled quantity at a specific impact energy in the case of mustard seeds, which also exhibit obvious charging effects and repulsion. Although the parameters such as mass per grain and packing density for the case of mustard seeds are intermediate between those for rice and cream of wheat, the spill quantity for comparable impact energy is considerably higher. These data will be presented and discussed.

Tir Is Essential for the Recruitment of Tks5 to Enteropathogenic Escherichia coli Pedestals

DEFF Research Database (Denmark)

Jensen, Helene Halkjær; Pedersen, Hans Nymand; Stenkjær, Eva

2015-01-01

Abstract Enteropathogenic Escherichia coli (EPEC) is a bacterial pathogen that infects the epithelial lining of the small intestine and causes diarrhea. Upon attachment to the intestinal epithe- lium, EPEC uses a Type III Secretion System to inject its own high affinity receptor Translo- cated in...

The affects of contrast medium on renal function in selective coronary angiography and intervention

International Nuclear Information System (INIS)

Chen Yueguang; Lv Baojing

2006-01-01

Selective coronary angiography and intervention with injection of contrast medium into the coronary arteries has become very common in dealing with coronary cardiac diseases. The excretion of contrast medium through kidneys may lead to acute renal functional insufficiency, especially for those suffering from chronic nephropathy, diabetes and cardiac functional disorder to form the so called 'contrast medium nephropathy' which is considered as the number second drug induced acute renal functional failure. Although routine preventive measure including low osmotic contrast medium and fine hydrotherapy have been taken, 14% incidences still occur with renal functional damage. The majority could be reversible but the minority needs emergent hemodialysis or even with persistent renal functional damage in a few ones. (authors)

El Naschie's coherence on the subquantum medium

International Nuclear Information System (INIS)

Agop, M.; Ioannou, P.D.; Nica, P.; Galusca, G.; Stefan, M.

2005-01-01

In the hydrodynamic formulation of the Scale Relativity theory one shows that a stable vortices distribution of bipolaron type induces superconducting pairs by means of the quantum potential. One builds the superconducting fractal by an iterated map and demonstrates that the superconducting pairs results as projections of this fractal. Thus, usual mechanisms (as example the exchange interaction used in the bipolaron theory) are reduced to the coherence on the subquantum medium in a ε (∞) space (El Naschie's coherence)

Effect of phenobarbital pretreatment on benzene biotransformation in the rat. Pt. 2. 9. 000 g supernatant and isolated perfused liver versus living rat

Energy Technology Data Exchange (ETDEWEB)

Gut, I.; Hatle, K.; Zizkova, L.

1981-03-01

Factors responsible for different quantitative effect of phenobarbital (PB) pretreatment on benzene metabolism to phenol in vivo and in vitro were studied in male Wistar rats. A more than 4-fold increase of benzene metabolism was observed with 9,000 g supernatant of liver homogenate, 2.8- to 4-fold increase with isolated perfused liver; phenol formation in vivo after oral benzene was increased by PB 2-fold, but only shortly following benzene administration and the enhancement rapidly diminished to 1.15-fold increase in the total excreted phenol. Benzene concentrations in 9,000 g supernatant incubations were 2 mM, those with isolated perfused livers were up to 4 mM, but those in blood in vivo were below 0.3 mM; the effect of PB induction in vivo disappeared along with decreasing benzene and increasing phenol blood concentrations which surpassed benzene 2-3 h after oral benzene administration. The effect of benzene concentration on the manifestation of PB induction is also supported by almost a 2-fold increased phenol formation in PB rats over controls in vivo after repeated administration of benzene. The elimination of radioactive metabolites of orally administered benzene-/sup 14/C, in urine was markedly inhibited by intraperitoneal administration of phenol, but not by pyrocatechol, resorcinol or hydroquinol suggesting that phenol might inhibit benzene metabolism in vivo especially when its concentration exceeds that of benzene.

Live and Heat-Killed Lactobacillus rhamnosus ATCC 7469 May Induce Modulatory Cytokines Profiles on Macrophages RAW 264.7.

Science.gov (United States)

Jorjão, Adeline Lacerda; de Oliveira, Felipe Eduardo; Leão, Mariella Vieira Pereira; Carvalho, Cláudio Antonio Talge; Jorge, Antonio Olavo Cardoso; de Oliveira, Luciane Dias

2015-01-01

This study aimed to evaluate the capacity of Lactobacillus rhamnosus and/or its products to induce the synthesis of cytokines (TNF-α, IL-1β, IL-4, IL-6, IL-10, and IL-12) by mouse macrophages (RAW 264.7). Three microorganism preparations were used: live L. rhamnosus (LLR) suspension, heat-killed L. rhamnosus (HKLR) suspension, and the supernatant of a heat-killed L. rhamnosus (SHKLR) suspension, which were cultured with macrophages (37°C, 5% CO2) for 2 h and 30 min. After that, cells were cultured for 16 h. The supernatants were used for the quantitation of cytokines, by ELISA. The results were compared with the synthesis induced by lipopolysaccharide (LPS) and analysed, using ANOVA and Tukey test, 5%. LLR and HKLR groups were able to significantly increase the production of TNF-α, IL-6, and IL-10 (P 0.05). All the L. rhamnosus suspensions were not able to produce detectable levels of IL-1β or significant levels of IL-4 and IL-12 (P > 0.05). In conclusion, live and heat-killed L. rhamnosus suspensions were able to induce the synthesis of different cytokines with proinflammatory (TNF-α and IL-6) or regulatory (IL-10) functions, suggesting the role of strain L. rhamnosus ATCC 7469 in the modulation or in the stimulation of immune responses.

Live and Heat-Killed Lactobacillus rhamnosus ATCC 7469 May Induce Modulatory Cytokines Profiles on Macrophages RAW 264.7

Directory of Open Access Journals (Sweden)

Adeline Lacerda Jorjão

2015-01-01

Full Text Available This study aimed to evaluate the capacity of Lactobacillus rhamnosus and/or its products to induce the synthesis of cytokines (TNF-α, IL-1β, IL-4, IL-6, IL-10, and IL-12 by mouse macrophages (RAW 264.7. Three microorganism preparations were used: live L. rhamnosus (LLR suspension, heat-killed L. rhamnosus (HKLR suspension, and the supernatant of a heat-killed L. rhamnosus (SHKLR suspension, which were cultured with macrophages (37°C, 5% CO2 for 2 h and 30 min. After that, cells were cultured for 16 h. The supernatants were used for the quantitation of cytokines, by ELISA. The results were compared with the synthesis induced by lipopolysaccharide (LPS and analysed, using ANOVA and Tukey test, 5%. LLR and HKLR groups were able to significantly increase the production of TNF-α, IL-6, and IL-10 (P0.05. All the L. rhamnosus suspensions were not able to produce detectable levels of IL-1β or significant levels of IL-4 and IL-12 (P>0.05. In conclusion, live and heat-killed L. rhamnosus suspensions were able to induce the synthesis of different cytokines with proinflammatory (TNF-α and IL-6 or regulatory (IL-10 functions, suggesting the role of strain L. rhamnosus ATCC 7469 in the modulation or in the stimulation of immune responses.

Construction of recombinant plasmid pIRESEgr-IFN γ and its expression in Lewis lung carcinoma induced by irradiation

International Nuclear Information System (INIS)

Yang Wei; Li Xiuyi; Gong Shouliang; Sun Ting; Gong Pingsheng

2007-01-01

Objective: To construct the recombinant plasmid pIRESEgr-IFN γ and detect its expression in Lewis lung carcinoma induced by irradiation in vitro. Methods: The recombinant plasmid pIRESEgr-IFN γ containing Egr-1 promoter and IFN γ gene was constructed with gene recombinant technique. The plasmid was transferred into Lewis lung carcinoma by liposome in vitro. The correlations of dose- and time-effects in the expression of IFN γ gene induced by X-ray were detected by ELISA. Results: The identification with enzymes proved that Egr-1 promoter and IFN γ gene were inserted into vector pIRESlneo correctly. After X-ray irradiation with different doses, the expression of IFN γ in the supernatant of Lewis lung carcinoma transfected by pIRESEgr-IFN γ was significantly higher than that in 0 Gy group (P<0.001). After 5 Gy X-ray irradiation, the expression of IFN γ was the highest, being 4.39 times as much as that in 0 Gy group. The expression of IFN γ in the supernatant increased after 5 Gy X-ray irradiation, being 6.27 times as much as that in 0 h group 36 h after irradiation. Conclusion: The recombinant plasmid pIRESEgr-IFN γ is constructed successfully, and it has the property of enhancing the expression of IFN γ gene induced by irradiation. (authors)

Effect of pH on radiation-induced apoptosis

International Nuclear Information System (INIS)

Chang, W. Song; Park, Heon J.; Lyons, John C.; Auger, Elizabeth A.; Lee, Hyung-Sik

1996-01-01

Purpose/Objective: The effect of environmental pH on the radiation-induced apoptosis in tumor cells in vitro was investigated. Materials and Methods: SCK mammary adenocarcinoma cells of A/J mice were irradiated with γ-rays using a 137 Cs irradiator and incubated in media of different pHs. After incubation at 37 degree sign C for 24-120 hrs., the extent of apoptosis was determined using agarose gel electrophoresis of DNA, in situ TUNEL staining, flow cytometry, and release of 3 H from 3 H-thymidine labeled cells. The membrane integrity, using the trypan blue exclusion method, and the clonogenicity of the cells were also determined. Results: Irradiation with 2-12 Gy of γ-rays induced apoptosis in pH 7.5 medium within 48 hrs. The radiation-induced apoptosis progressively declined as the medium pH was lowered so that little apoptosis occurred in 48 hrs. after irradiation with 12 Gy in pH 6.6 medium. However, when the cells were irradiated and incubated for 48 hrs. in pH 6.6 medium and then medium was replaced with pH 7.5 medium, apoptosis promptly occurred. Apoptosis also occurred even in pH 6.6 medium when the cells were irradiated and maintained in pH 7.5 medium for 8 hrs. or longer post-irradiation before incubation in pH 6.6 medium. Conclusion: An acidic environment markedly suppresses radiation-induced apoptosis probably by suppressing the expression of initial signals responsible for irradiation-induced apoptosis. Indications are that the signals persist in an acidic environment and trigger apoptosis when the environmental acidity is eased. Our results suggest that the acidic environment in human tumors may inhibit the apoptosis after irradiation. However, apoptosis may be triggered when reoxygenation occurs after irradiation, and thus, the intratumor environment becomes less acidic after irradiation. Not only the change in pO 2 but the change in pH during the course of fractionated radiotherapy may greatly influence the outcome of the treatment

Contrast medium-induced nephropathy: the pathophysiology

DEFF Research Database (Denmark)

Persson, P B; Tepel, Martin

2006-01-01

A widespread, rather general, definition of contrast-induced nephropathy (CIN) is an impairment in renal function occurring within 3 days following the intravascular administration of contrast media (CM) and the absence of an alternative aetiology. In spite of the vast clinical importance of CIN...... haemodynamics, regional hypoxia, auto-, and paracrine factors (adenosine, endothelin, reactive oxygen species) to direct cytotoxic effects. Although these potential mediators of CIN will be discussed separately, several factors may act in concert to perturb kidney function after exposure to contrast media. From...... the current knowledge of the mechanisms causing CIN, it is not possible to recommend a certain class of contrast media, except to avoid large doses of CM of the first generation. From a pathophysiological perspective, volume expansion is effective in avoiding CIN, since water permeability of the collecting...

L-Sorbose but not D-tagatose induces hemolysis of dog erythrocytes in vitro.

Science.gov (United States)

Bär, A; Leeman, W R

1999-04-01

Previous investigations have demonstrated that L-sorbose induces hemolysis of dog erythrocytes. This effect is probably the consequence of an ATP depletion of the red blood cells subsequent to inhibition of hexokinase, and thus the glycolytic pathway, by sorbose 1-phosphate. In the present study, the susceptibility of dog erythrocytes to D-tagatose, a stereoisomer of L-sorbose, was examined. Washed dog erythrocytes were suspended in Hanks' balanced salt solution (HBSS, containing 5.6 mM glucose) with or without the addition of 0.6, 6, and 60 mM L-sorbose or D-tagatose, or in HBSS with total glucose concentrations of 5.6, 6 and 60 mM D-glucose. After incubation for 24 h at 34 degrees C, the suspensions were centrifuged, and the percentage of hemolysis was determined by measuring the hemoglobin in the sediment and the supernatant. The amount of hemoglobin released in the medium did not differ significantly between the control (HBSS) and the test incubations with glucose or D-tagatose supplementation. In contrast, the addition of 6 and 60 mM L-sorbose resulted in significant hemolysis. At the low dose (0.6 mM), L-sorbose did not have an adverse effect. It is concluded that D-tagatose, unlike L-sorbose, does not have a hemolytic effect on canine erythrocytes. Copyright 1999 Academic Press.

Recovering and recycling uranium used for production of molybdenum-99

Science.gov (United States)

Reilly, Sean Douglas; May, Iain; Copping, Roy; Dale, Gregory Edward

2017-12-12

A processes for recycling uranium that has been used for the production of molybdenum-99 involves irradiating a solution of uranium suitable for forming fission products including molybdenum-99, conditioning the irradiated solution to one suitable for inducing the formation of crystals of uranyl nitrate hydrates, then forming the crystals and a supernatant and then separating the crystals from the supernatant, thus using the crystals as a source of uranium for recycle. Molybdenum-99 is recovered from the supernatant using an adsorbent such as alumina. Another process involves irradiation of a solid target comprising uranium, forming an acidic solution from the irradiated target suitable for inducing the formation of crystals of uranyl nitrate hydrates, then forming the crystals and a supernatant and then separating the crystals from the supernatant, thus using the crystals as a source of uranium for recycle. Molybdenum-99 is recovered from the supernatant using an adsorbent such as alumina.

Comparison of Growth Kinetics of Various Pathogenic E. coli on Fresh Perilla Leaf

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Juhui Kim

2013-08-01

Full Text Available Growth kinetics for Escherichia coli O157:H7 in perilla leaves were compared to those of pathogenic E. coli strains, including enteropathogenic (EPEC, enterotoxigenic (ETEC, enteroinvasive (EIEC and other enterohemorrhagic (EHEC at 13, 17, 24, 30 and 36 °C. Models for lag time (LT, specific growth rate (SGR and maximum population density (MPD as a function of temperature were developed. The performance of the models was quantified using the ratio method and an acceptable prediction zone method. Significant differences in SGR and LT among the strains were observed at all temperatures. Overall, the shortest LT was observed with E. coli O157:H7, followed by EPEC, other EHEC, EIEC and ETEC, while the fastest growth rates were noted in EPEC, followed by E. coli O157:H7, ETEC, other EHEC and EIEC. The models for E. coli O157:H7 in perilla leaves was suitable for use in making predictions for EPEC and other EHEC strains.

Genomic comparison of Escherichia coli serotype O103:H2 isolates with and without verotoxin genes: implications for risk assessment of strains commonly found in ruminant reservoirs

Directory of Open Access Journals (Sweden)

Robert Söderlund

2016-02-01

Full Text Available Introduction: Escherichia coli O103:H2 occurs as verotoxigenic E. coli (VTEC carrying only vtx1 or vtx2 or both variants, but also as vtx-negative atypical enteropathogenic E. coli (aEPEC. The majority of E. coli O103:H2 identified from cases of human disease are caused by the VTEC form. If aEPEC strains frequently acquire verotoxin genes and become VTEC, they must be considered a significant public health concern. In this study, we have characterized and compared aEPEC and VTEC isolates of E. coli O103:H2 from Swedish cattle. Methods: Fourteen isolates of E. coli O103:H2 with and without verotoxin genes were collected from samples of cattle feces taken during a nationwide cattle prevalence study 2011–2012. Isolates were sequenced with a 2×100 bp setup on a HiSeq2500 instrument producing >100× coverage per isolate. Single-nucleotide polymorphism (SNP typing was performed using the genome analysis tool kit (GATK. Virulence genes and other regions of interest were detected. Susceptibility to transduction by two verotoxin-encoding phages was investigated for one representative aEPEC O103:H2 isolate. Results and Discussion: This study shows that aEPEC O103:H2 is more commonly found (64% than VTEC O103:H2 (36% in the Swedish cattle reservoir. The only verotoxin gene variant identified was vtx1a. Phylogenetic comparison by SNP analysis indicates that while certain subgroups of aEPEC and VTEC are closely related and have otherwise near identical virulence gene repertoires, they belong to separate lineages. This indicates that the uptake or loss of verotoxin genes is a rare event in the natural cattle environment of these bacteria. However, a representative of a VTEC-like aEPEC O103:H2 subgroup could be stably lysogenized by a vtx-encoding phage in vitro.

Characteristics of diarrheagenic Escherichia coli among children under 5 years of age with acute diarrhea: a hospital based study.

Science.gov (United States)

Zhou, Yu; Zhu, Xuhui; Hou, Hongyan; Lu, Yanfang; Yu, Jing; Mao, Lie; Mao, Liyan; Sun, Ziyong

2018-02-01

Diarrhea is the leading infectious cause of childhood morbidity and mortality. Among bacterial agents, diarrheagenic Escherichia coli (DEC) is the major causal agent of childhood diarrhea in developing countries, particularly in children under the age of 5 years. Here, we performed a hospital-based prospective study to explore the pathotype distribution, epidemiological characteristics and antibiotic resistance patterns of DEC from coli (EPEC) isolates were identified simultaneously with serology. Furthermore, antimicrobial sensitivity tests and sequencing of antibiotic resistance-related genes were conducted. DEC strains were identified in 7.9% of the 684 stool samples. Among them, the most commonly detected pathotype was EPEC (50.0% of DEC), of which 77.8% were classified as atypical EPEC (aEPEC). Age and seasonal distribution revealed that DEC tended to infect younger children and to occur in summer/autumn periods. Multidrug-resistant DEC isolates were 66.7%; resistance rates to ampicillin, co-trimoxazole, cefazolin, cefuroxime, cefotaxime, and ciprofloxacin were ≥ 50%. Among 5 carbapenem-resistant DEC, 60.0% were positive for carbapenemase genes (2 blaNDM-1 and 1 blaKPC-2). Among 30 cephalosporin-resistant DEC, 93.3% were positive for extended-spectrum β-lactamase (ESBL) genes, with blaTEM-1 and blaCTX-M-55 being the most common types. However, no gyrA or gyrB genes were detected in 16 quinolone-resistant isolates. Notably, aEPEC, which has not received much attention before, also exhibited high rates of drug resistance (81.0%, 66.7%, and 14.3% for ampicillin, co-trimoxazole , and carbapenem resistance, respectively). EPEC was the most frequent DEC pathotype in acute diarrheal children, with aEPEC emerging as a dominant diarrheal agent in central China. Most DEC strains were multidrug-resistant, making even ciprofloxacin unsuitable for empiric treatment against DEC infection. Among carbapenem-resistant DEC strains, those harboring blaNDM-1 and blaKPC-2

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

Science.gov (United States)

Rick, Jochen; Lamberty, K. K.

2005-01-01

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

Study on MCP-1 related to inflammation induced by biomaterials

International Nuclear Information System (INIS)

Ding Tingting; Sun Jiao; Zhang Ping

2009-01-01

The study of inflammation is important for understanding the reaction between biomaterials and the human body, in particular, the interaction between biomaterials and immune system. In the current study, rat macrophages were induced by multiple biomaterials with different biocompatibilities, including polyvinyl chloride (PVC) containing 8% of organic tin, a positive control material with cellular toxicity. Human umbilical vein endothelial cells (ECV-304), cultured with PRMI-1640, were detached from cells cultured with the supernatant of macrophages containing TNF-α and IL-1β because of stimulation by biomaterials. The cells were then treated with different biomaterials. Then both TNF-α and IL-1β in macrophages were detected by ELISA. Levels of monocyte chemoattractant protein-1 (MCP-1) were measured by RT-PCR. The results suggested that the expression of TNF-α and IL-1β was elevated by polytetrafluoroethylene (PTFE), polylactic-co-glycolic acid (PLGA) and American NPG alloy (p < 0.001). The level of MCP-1 cultured in supernatant of macrophages was higher than in PRMI-1640 with the same biomaterials. And the exposure to PTFE, PLGA and NPG resulted in the high expression of MCP-1 (p < 0.001) following cytokine stimulation. MCP-1 was also significantly expressed in β-tricalcium phosphate (β-TCP) and calcium phosphate cement samples (CPC) (p < 0.01). Thus, TNF-α, IL-1β and MCP-1 had played an important role in the immune reaction induced by biomaterials and there was a close relationship between the expression of cytokines and biomcompatibility of biomaterials. Furthermore, these data suggested that MCP-1 was regulated by TNF-α and IL-1β, and activated by both cytokines and biomaterials. The data further suggested that the expression of MCP-1 could be used as a marker to indicate the degree of immune reaction induced by biomaterials.

Linear and nonlinear light propagations in a Doppler-broadened medium via electromagnetically induced transparency

International Nuclear Information System (INIS)

Li Liang; Huang Guoxiang

2010-01-01

We present a systematic theoretical study to deal with linear and nonlinear light propagations in a Doppler-broadened three-level Λ system via electromagnetically induced transparency (EIT), with incoherent population exchange between two lower energy levels taken into account. Through a careful analysis of base state and linear excitation, we show that the EIT condition of the system is given by |Ω c | 2 γ 31 >>2γ 21 Δω D 2 , where Ω c is half the Rabi frequency of the control field, Δω D is the Doppler width, and γ jl is the decay rate of the coherence between states |j> and |l>. Under this condition, the effect of incoherent population exchange is insignificant, while dephasing dominates the decoherence of the system. This condition also ensures the validity of the weak nonlinear perturbation theory used in this work for solving the Maxwell-Bloch equations with inhomogeneous broadening. We then investigate the nonlinear propagation of the probe field and show that it is possible to form temporal optical solitons in the Doppler-broadened medium. Such solitons have ultraslow propagating velocity and can be generated in very low light power. The possibility of realizing (1+1)-dimensional and (2+1)-dimensional spatial optical solitons in the adiabatic regime of the system is also discussed.

Studies of the small bowel surface by scanning electron microscopy in infants with persistent diarrhea

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U. Fagundes-Neto

2000-12-01

Full Text Available We describe the ultrastructural abnormalities of the small bowel surface in 16 infants with persistent diarrhea. The age range of the patients was 2 to 10 months, mean 4.8 months. All patients had diarrhea lasting 14 or more days. Bacterial overgrowth of the colonic microflora in the jejunal secretion, at concentrations above 10(4 colonies/ml, was present in 11 (68.7% patients. The stool culture was positive for an enteropathogenic agent in 8 (50.0% patients: for EPEC O111 in 2, EPEC O119 in 1, EAEC in 1, and Shigella flexneri in 1; mixed infections due to EPEC O111 and EAEC in 1 patient, EPEC O119 and EAEC in 1 and EPEC O55, EPEC O111, EAEC and Shigella sonnei in 1. Morphological abnormalities in the small bowel mucosa were observed in all 16 patients, varying in intensity from moderate 9 (56.3% to severe 7 (43.7%. The scanning electron microscopic study of small bowel biopsies from these subjects showed several surface abnormalities. At low magnification (100X most of the villi showed mild to moderate stunting, but on several occasions there was subtotal villus atrophy. At higher magnification (7,500X photomicrographs showed derangement of the enterocytes; on several occasions the cell borders were not clearly defined and very often microvilli were decreased in number and height; in some areas there was a total disappearance of the microvilli. In half of the patients a mucus-fibrinoid pseudomembrane was seen partially coating the enterocytes, a finding that provides additional information on the pathophysiology of persistent diarrhea.

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

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Shahrul Hisham Zainal Ariffin

2013-01-01

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

Optimization of laccase production by two strains of Ganoderma lucidum using phenolic and metallic inducers

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Francisco Kuhar

Full Text Available Ganoderma lucidum (Curtis P. Karst is a white rot fungus that is able to degrade the lignin component in wood. The ability of two strains of this species to produce the ligninolytic enzyme laccase was assessed. After the evaluation of induction with heavy metals and phenolic compounds, it was found that among the tested substances, copper and ferulic acid are the best laccase inducers. It was also observed that the two types of inducers (phenolic and metallic produce different electrophoretic patterns of laccase activity. Optimized concentrations of inducers were obtained through a factorial design and the thermal stability of optimized supernatants was studied at a wide range of acidic pH. We found that the enzyme is more thermostable at higher pH values.

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

Science.gov (United States)

Enoksen, Anette Einan; Dickerson, Paul

2018-04-01

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

Dicty_cDB: Contig-U14921-1 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available 498 e-139 AF399172_1( AF399172 |pid:none) Eutintinnus pectinis clone Epec99a... 497 e-139 AF399156_1( AF399...AF399175_1( AF399175 |pid:none) Eutintinnus pectinis clone Epec99a... 496 e-138 AF399137_1( AF399137 |pid:no

Prototypical antipsychotic drugs protect hippocampal neuronal cultures against cell death induced by growth medium deprivation

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Williams Sylvain

2006-03-01

Full Text Available Abstract Background Several clinical studies suggested that antipsychotic-based medications could ameliorate cognitive functions impaired in certain schizophrenic patients. Accordingly, we investigated the effects of various dopaminergic receptor antagonists – including atypical antipsychotics that are prescribed for the treatment of schizophrenia – in a model of toxicity using cultured hippocampal neurons, the hippocampus being a region of particular relevance to cognition. Results Hippocampal cell death induced by deprivation of growth medium constituents was strongly blocked by drugs including antipsychotics (10-10-10-6 M that display nM affinities for D2 and/or D4 receptors (clozapine, haloperidol, (±-sulpiride, domperidone, clozapine, risperidone, chlorpromazine, (+-butaclamol and L-741,742. These effects were shared by some caspases inhibitors and were not accompanied by inhibition of reactive oxygen species. In contrast, (--raclopride and remoxipride, two drugs that preferentially bind D2 over D4 receptors were ineffective, as well as the selective D3 receptor antagonist U 99194. Interestingly, (--raclopride (10-6 M was able to block the neuroprotective effect of the atypical antipsychotic clozapine (10-6 M. Conclusion Taken together, these data suggest that D2-like receptors, particularly the D4 subtype, mediate the neuroprotective effects of antipsychotic drugs possibly through a ROS-independent, caspase-dependent mechanism.

Do medium heavy fragments give evidence for a liquid-gas phase transition

International Nuclear Information System (INIS)

Trockel, R.; Hildenbrand, K.D.; Lynen, U.; Mueller, W.F.J.; Rabe, H.J.; Sann, H.; Stelzer, H.; Wada, R.; Brummund, N.; Glasow, R.; Kampert, K.H.; Santo, R.; Pelte, D.; Pochodzalla, J.; Eckert, E.

1985-09-01

Light and medium heavy fragments have been measured in light ion induced reactions at intermediate energies. The energy spectra have been parametrized with moving source fits. The resulting temperatures and yields do not confirm the expectations of a liquid-gas phase transition. (orig.)

Vascular endothelial growth factor (VEGF), produced by feline infectious peritonitis (FIP) virus-infected monocytes and macrophages, induces vascular permeability and effusion in cats with FIP.

Science.gov (United States)

Takano, Tomomi; Ohyama, Taku; Kokumoto, Aiko; Satoh, Ryoichi; Hohdatsu, Tsutomu

2011-06-01

Feline infectious peritonitis virus (FIPV) causes a fatal disease called FIP in Felidae. The effusion in body cavity is commonly associated with FIP. However, the exact mechanism of accumulation of effusion remains unclear. We investigated vascular endothelial growth factor (VEGF) to examine the relationship between VEGF levels and the amounts of effusion in cats with FIP. Furthermore, we examined VEGF production in FIPV-infected monocytes/macrophages, and we used feline vascular endothelial cells to examine vascular permeability induced by the culture supernatant of FIPV-infected macrophages. In cats with FIP, the production of effusion was related with increasing plasma VEGF levels. In FIPV-infected monocytes/macrophages, the production of VEGF was associated with proliferation of virus. Furthermore, the culture supernatant of FIPV-infected macrophages induced hyperpermeability of feline vascular endothelial cells. It was suggested that vascular permeability factors, including VEGF, produced by FIPV-infected monocytes/macrophages might increase the vascular permeability and the amounts of effusion in cats with FIP. Copyright © 2011 Elsevier B.V. All rights reserved.

Borel sum rules for octet baryons in nuclear medium

International Nuclear Information System (INIS)

Kondo, Y.; Morimatsu, O.

1992-06-01

Borel sum rules are examined for octet baryons in the nuclear medium. First, it is noticed that in the medium the dispersion relation is realized for the retarded correlation Π R (ω, q 2 ) in the energy ω. Then, Π R (ω, q 2 ) is split into even and odd parts of ω in order to apply the Borel transformation. The obtained Borel sum rules differ from those of previous works. The mass shifts of octet baryons are calculated in the leading order of the operator product expansion with linear density approximation for the condensates. It is found that both scalar and vector condensates of the quark field, and + q>, induce attraction to the octet baryons in the medium in contrast to the results of previous works. It is also found that |δM N | > |δM Λ | > |δM Σ | ∼ |δM Ξ |. The absolute values, however, turn out to be one order of magnitude larger than those empirically known if a Borel mass of around 1 GeV is used in the present approximation. (author)

Jet formation and interference in a thin QCD medium

CERN Document Server

Casalderrey-Solana, Jorge; Tywoniuk, Konrad

2016-11-29

In heavy-ion collisions, an abundant production of high-energy QCD jets allows to study how these multiparticle sprays are modified as they pass through the quark-gluon plasma. In order to shed new light on this process, we compute the inclusive two-gluon rate off a hard quark propagating through a color deconfined medium at first order in medium opacity. We explicitly impose an energy ordering of the two emitted gluons, such that the "hard" gluon can be thought of as belonging to the jet substructure while the other is a "soft" emission (which can be collinear or medium-induced). Our analysis focusses on two specific limits that clarify the modification of the additional angle- and formation time-ordering of splittings. In one limit, the formation time of the "hard" gluon is short compared to the "soft" gluon formation time, leading to a probabilistic formula for production of and subsequent radiation off a quark-gluon antenna. In the other limit, the ordering of formation is reverted, which automatically le...

[Effects of non-saccharomyces albicans metabolic products on the proliferation of human umbilical vein endothelial cell ECV304].

Science.gov (United States)

Chen, Bin; Che, Tuanjie; Bai, Decheng; He, Xiangyi

2013-04-01

To evaluate the effects of non-Saccharomyces albicans metabolic products on the cell cycle distribution and proliferation of human umbilical vein endothelial cell ECV304 cells in vitro. The parallel dilution supernatant of Saccharomyces tropicalis, Saccharomyces krusei and Saccharomyces glabrata were prepared, and 1, 4, 16-fold(s) diluted concentration and control group were set up. The line of human umbilical vein endothelial cell ECV304 was cultured in vitro and treated by non-Saccharomyces albicans supernatant. The proliferous effect of ECV304 induced by non-Saccharomyces albicans supernatant after 24, 48, 72 h was detected by the methods of MTT, and the changes of cell density and cycle after 48 h were investigated by inverted microscope and flow cytometry. At the 24th hour, all of the higher concentration (1-fold) of non-Saccharomyces albicans supernatant and the 4-folds diluted Saccharomyces krusei could promote ECV304 proliferation(P Saccharomyces albicans supernatant at 48h and 72th hour, Saccharomyces krusei supernatant and Saccharomyces glabrata supernatant significantly increased proliferation rate of ECV304, while Saccharomyces tropicalis supernatant group showed no significant change no matter which concentration was tested. At 48th hour after adding the non-Saccharomyces albicans supernatant, the ECV304 cells density treated by Saccharomyces krusei supernatant and Saccharomyces glabrata supernatant were significantly higher under the inverted microscope. The G0/G1 population of ECV304 cells decreased while cell proliferation index (PI) increased after incubated with Saccharomyces krusei supernatant and Saccharomyces glabrata supernatant for 48 hours (P Saccharomyces tropicalis group showed no significant change (P > 0.05). The metabolic products of Sacharoymces krusei and Saccharomyces glabrata could induce proliferation of ECV304 cell, which suggests non-Saccharomyces albicans should be undergone more attention clinically in detection and treatment.

Sulfite induces release of lipid mediators by alveolar macrophages

Energy Technology Data Exchange (ETDEWEB)

Beck-Speier, I.; Dayal, N.; Maier, L. [GSF - National Research Center for Environment and Health, Neuherberg (Germany). Inst. for Inhalation Biology; Denzlinger, C. [Tuebingen Univ. (Germany). Dept. II, Medical Clinic; Haberl, C. [Tuebingen Univ. (Germany). Dept. III, Medical Clinic

1998-03-01

Air pollutants are supposed to modulate physiological responses of alveolar macrophages (AM). This study was addressed to the question whether at neutral pH sulfur(IV) species in comparison to sulfur(VI) species cause AM to release proinflammatory mediators and which pathways are involved in their generation. Supernatants obtained from canine AM treated with sulfite (0.1 mM to 2 mM) enhanced the respiratory burst of canine neutrophils, measured by lucigenin-dependent chemiluminescence, whereas supernatants derived from AM treated with sulfate (1 mM) did not. The neutrophil-stimulating activity released by sulfite-treated AM consisted of platelet-activating factor (PAF) and leukotriene B{sub 4} (LTB{sub 4}) as shown by desensitization of the platelet-activating factor (PAF) and leukotriene B{sub 4} (LTB{sub 4}) as shown by desensitization of the corresponding receptors. Inhibitors of phospholipase A{sub 2} substantially suppressed release of neutrophil-stimulating activity by sulfite-treated AM. Inhibition of 5-lipoxygenase in sulfite-treated AM also reduced neutrophil-stimulating activity, while inhibition of cyclooxygenase had no effect. In conclusion, sulfite induces AM to release lipid mediators via phospholipase A{sub 2}- and 5-lipoxygenase-dependent pathways. These mediators activate neutrophils via the receptors for PAF and LTB{sub 4}. (orig.)

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

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Hiroyuki Sanjo

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

Cutinase production by Fusarium oxysporum in liquid medium using central composite design.

Science.gov (United States)

Pio, Tatiana Fontes; Macedo, Gabriela Alves

2008-01-01

The objective of the present study was to measure the production of cutinase by Fusarium oxysporum in the presence of several carbon and nitrogen sources (glycides, fatty acids and oils, and several organic and inorganic nitrogen sources), trying to find a cost-effective substitute for cutin in the culture medium as an inducer of cutinase production. The results were evaluated by the Tukey test, and flaxseed oil was found to give the best results as a cutinase inducer. The authors optimized the composition of the growth medium employing response surface methodology. The experimental results were fitted to a second-order polynomial model at a 95% level of significance (p < 0.05). The greatest cutinolytic activity was obtained in a liquid mineral medium supplemented with flaxseed oil, showing an increase in enzymatic activity from 11 to 22.68 U/mL after 48 h of fermentation. A CCD study of the fermentation conditions was carried out, and the best production of cutinase was registered with the use of 30 degrees C and 100 rpm. These results support the use of flaxseed oil as a substitute for cutin, which is difficult and expensive to obtain, for the production of cutinase in a larger scale.

Lymphocyte-conditioned medium in combination with interleukin-2 effectively induces antitumour autoimmunity by adoptive transfer of short activated killer (SHAK) cells.

Science.gov (United States)

Buer, J; Hilse, R; Dallmann, I; Grosse, J; Kirchner, H; Zorn, U; Hänninen, E L; Franzke, A; Duensing, S; Poliwoda, H

1995-03-01

In this study, effective antitumour immunity was transferred by autologous short activated killer (SHAK) cells induced over four hours with lymphocyte conditioned medium (LCM) and recombinant interleukin-2 (rIL-2). Among eight patients with progressive metastatic renal cell carcinoma refractory to standard therapy, there were six objective tumour responses to SHAKs. Progression-free survival ranged from 0 to 8+ months, and overall survival ranged from 2 to 14+ months, with a median of 9+ months. Systemic toxicity of SHAKs was limited to flulike symptoms. Patient SHAKs provided a tumour-specific immunity, both cellular and humoral (expression and secretion of secondary cytokines, including IL-2, GM-CSF, INF-gamma and TNF-alpha), far superior to rIL-2 activated killer cells.

Nitric oxide-dependent vasorelaxation induced by extractive solutions and fractions of Maytenus ilicifolia Mart ex Reissek (Celastraceae) leaves.

Science.gov (United States)

Rattmann, Yanna D; Cipriani, Thales R; Sassaki, Guilherme L; Iacomini, Marcello; Rieck, Lia; Marques, Maria C A; da Silva-Santos, José E

2006-04-06

This study reveals that an ethanolic supernatant obtained from an aqueous extractive solution prepared from residues of methanolic extracts of ground leaves of Maytenus ilicifolia is able to cause a concentration- and endothelium-dependent relaxation in pre-contract rat aorta rings, with EC(50) of 199.7 (190-210) microg/ml. The non-selective nitric oxide synthase inhibitors l-NAME and l-NMMA abolished this effect, while superoxide dismutase and MnTBAP (a non-enzymatic superoxide dismutase mimetic) enhanced it. Further, relaxation induced by this ethanolic supernatant have been strongly inhibited by the guanylate cyclase inhibitors methylene blue and ODQ, as well as by the potassium channel blockers 4-aminopyridine and tetraethylammonium, but was unchanged by the cyclooxygenase inhibitor indomethacin and the membrane receptor antagonists atropine, HOE-140 and pirilamine. Partition of the ethanolic supernatant between H(2)O and EtOAc generated a fraction several times more potent, able to fully relax endothelium-intact aorta rings with an EC(50) of 4.3 (3.9-4.8) microg/ml. (13)C NMR spectrum of this fraction showed signals typical of catechin. This study reveals that the leaves of M. ilicifolia possess one or more potent substances able to relax endothelium-intact rat aorta rings, an event that appears to involve nitric oxide production, guanylate cyclase activation and potassium channel opening.

Optimized exosome isolation protocol for cell culture supernatant and human plasma

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Richard J. Lobb

2015-07-01

Full Text Available Extracellular vesicles represent a rich source of novel biomarkers in the diagnosis and prognosis of disease. However, there is currently limited information elucidating the most efficient methods for obtaining high yields of pure exosomes, a subset of extracellular vesicles, from cell culture supernatant and complex biological fluids such as plasma. To this end, we comprehensively characterize a variety of exosome isolation protocols for their efficiency, yield and purity of isolated exosomes. Repeated ultracentrifugation steps can reduce the quality of exosome preparations leading to lower exosome yield. We show that concentration of cell culture conditioned media using ultrafiltration devices results in increased vesicle isolation when compared to traditional ultracentrifugation protocols. However, our data on using conditioned media isolated from the Non-Small-Cell Lung Cancer (NSCLC SK-MES-1 cell line demonstrates that the choice of concentrating device can greatly impact the yield of isolated exosomes. We find that centrifuge-based concentrating methods are more appropriate than pressure-driven concentrating devices and allow the rapid isolation of exosomes from both NSCLC cell culture conditioned media and complex biological fluids. In fact to date, no protocol detailing exosome isolation utilizing current commercial methods from both cells and patient samples has been described. Utilizing tunable resistive pulse sensing and protein analysis, we provide a comparative analysis of 4 exosome isolation techniques, indicating their efficacy and preparation purity. Our results demonstrate that current precipitation protocols for the isolation of exosomes from cell culture conditioned media and plasma provide the least pure preparations of exosomes, whereas size exclusion isolation is comparable to density gradient purification of exosomes. We have identified current shortcomings in common extracellular vesicle isolation methods and provide a

[Prognostic estimation in critical patients. Validation of a new and very simple system of prognostic estimation of survival in an intensive care unit].

Science.gov (United States)

Abizanda, R; Padron, A; Vidal, B; Mas, S; Belenguer, A; Madero, J; Heras, A

2006-04-01

To make the validation of a new system of prognostic estimation of survival in critical patients (EPEC) seen in a multidisciplinar Intensive care unit (ICU). Prospective analysis of a patient cohort seen in the ICU of a multidisciplinar Intensive Medicine Service of a reference teaching hospital with 19 beds. Four hundred eighty four patients admitted consecutively over 6 months in 2003. Data collection of a basic minimum data set that includes patient identification data (gender, age), reason for admission and their origin, prognostic estimation of survival by EPEC, MPM II 0 and SAPS II (the latter two considered as gold standard). Mortality was evaluated on hospital discharge. EPEC validation was done with analysis of its discriminating capacity (ROC curve), calibration of its prognostic capacity (Hosmer Lemeshow C test), resolution of the 2 x 2 Contingency tables around different probability values (20, 50, 70 and mean value of prognostic estimation). The standardized mortality rate (SMR) for each one of the methods was calculated. Linear regression of the EPEC regarding the MPM II 0 and SAPS II was established and concordance analyses were done (Bland-Altman test) of the prediction of mortality by the three systems. In spite of an apparently good linear correlation, similar accuracy of prediction and discrimination capacity, EPEC is not well-calibrated (no likelihood of death greater than 50%) and the concordance analyses show that more than 10% of the pairs were outside the 95% confidence interval. In spite of its ease of application and calculation and of incorporating delay of admission in ICU as a variable, EPEC does not offer any predictive advantage on MPM II 0 or SAPS II, and its predictions adapt to reality worse.

Contrast media induced acute renal failure in diabetics

International Nuclear Information System (INIS)

Rambausek, M.

1985-01-01

Dehydration, preexisting renal insufficiency, multiple myeloma and insulin-dependent diabetes mellitus are known risk factors for a radiocontrast medium induced acute renal failure. In 90% of patients with insulin-dependent diabetes mellitus, renal insufficiency and proteinuria, a further detoriation of renal function can be expected after i.v. administration of radiocontrast medium. Recent concepts on the genesis of acute renal failure after radiocontrast medium in multiple myeloma emphasize the role of tubular blocade (tubular precipitation of myeloma protein with contrast medium). In insulin-dependent diabetic patients we found altered carbohydrate composition of urinary Tamm Horsfall Protein (THP), with increased glucose and diminished N-acetyl-neuraminicacid content. This was paralleled by a difference in an in-vitro system of coprecipitation where THP of diabetes triggered more pronounced calcium dependent coprecipitation of contrast medium and albumin. These in-vitro findings might be important for the explanation of the genesis of radiocontrast medium-induced acute renal failure in insulin-dependent diabetes mellitus. (orig.) [de

Dynamical Model about Rumor Spreading with Medium

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Xiaxia Zhao

2013-01-01

Full Text Available Rumor is a kind of social remark, that is untrue, and not be confirmed, and spreads on a large scale in a short time. Usually, it can induce a cloud of pressure, anxiety, and panic. Traditionally, it is propagated by word of mouth. Nowadays, with the emergence of the internet, rumors can be spread by instant messengers, emails, or publishing. With this new pattern of spreading, an ISRW dynamical model considering the medium as a subclass is established. Beside the dynamical analysis of the model, we mainly explore the mechanism of spreading of individuals-to-individuals and medium-to-individual. By numerical simulation, we find that if we want to control the rumor spreading, it will not only need to control the rate of change of the spreader subclass, but also need to control the change of the information about rumor in medium which has larger influence. Moreover, to control the effusion of rumor is more important than deleting existing information about rumor. On the one hand, government should enhance the management of internet. On the other hand, relevant legal institutions for punishing the rumor creator and spreader on internet who can be tracked should be established. Using this way, involved authorities can propose efficient measures to control the rumor spreading to keep the stabilization of society and development of economy.

Development program for magnetically assisted chemical separation: Evaluation of cesium removal from Hanford tank supernatant

International Nuclear Information System (INIS)

Nunez, L.; Buchholz, B.A.; Ziemer, M.; Dyrkacz, G.; Kaminski, M.; Vandegrift, G.F.; Atkins, K.J.; Bos, F.M.; Elder, G.R.; Swift, C.A.

1994-12-01

Magnetic particles (MAG*SEP SM ) coated with various absorbents were evaluated for the separation and recovery of low concentrations of cesium from nuclear waste solutions. The MAG*SEP SM particles were coated with (1) clinoptilolite, (2) transylvanian volcanic tuff, (3) resorcinol formaldehyde, and (4) crystalline silico-titanate, and then were contacted with a Hanford supernatant simulant. Particles coated with the crystalline silico-titanate were identified by Bradtec as having the highest capacity for cesium removal under the conditions tested (variation of pH, ionic strength, cesium concentration, and absorbent/solution ratio). The MAG*SEP SM particles coated with resorcinol formaldehyde had high distribution ratios values and could also be used to remove cesium from Hanford supernant simulant. Gamma irradiation studies were performed on the MAG*SEP SM particles with a gamma dose equivalent to 100 cycles of use. This irradiation decreased the loading capacity and distribution ratios for the particles by greater than 75%. The particles demonstrated high sensitivity to radiolytic damage due to the degradation of the polymeric regions. These results were supported by optical microscopy measurements. Overall, use of magnetic particles for cesium separation under nuclear waste conditions was found to be marginally effective

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

Science.gov (United States)

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

2016-01-01

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

Caffeine and D2O medium interact in affecting the expression of radiation-induced potentially lethal damage

International Nuclear Information System (INIS)

Utsumi, H.; Elkind, M.M.

1991-01-01

Earlier work has been extended to compare the killing of long-phase V79 Chinese hamster cells by ionizing radiation when they are treated immediately after irradiation with medium containing either caffeine or 90% D 2 O. The object was to determine if the enhanced killing due to post-treatment with caffeine, or D 2 O, resulted from action on the same sector of potentially lethal damage as appeared to be the case for hypertonic shock and D 2 O medium. The treatments by themselves were not toxic to unirradiated cells. We found that the enhanced expression of potentially lethal damage by post-treatment with caffeine or D 2 O medium is similar. For example, the kinetic of the repair of the potentially lethal damage expressible by either post-treatment was similar, and an additive enhancement of potentially lethal damage occurred when the two treatments were administered sequentially. These findings suggest that caffeine and D 2 O medium affect the same sector of potentially lethal damage. When the two treatments were combined, however, they competed with each other. Thus, although caffeine and D 2 O medium act on the same sector of potentially lethal damage they do so differently, suggesting that more than one pathway of the expression of radiation damage can result in the same phenotypic effect. (author)

Antimicrobial activity of Bacillus amyloliquefaciens LBM 5006 is enhanced in the presence of Escherichia coli.

Science.gov (United States)

Benitez, Lisianne; Correa, AnaPaula; Daroit, Daniel; Brandelli, Adriano

2011-03-01

Increased antimicrobial activity was observed when Bacillus amyloliquefaciens LBM 5006 strain was cultivated in the presence of thermally inactivated cells of Escherichia coli, but not with Staphylococcus aureus, Listeria monocytogenes, or Bacillus cereus. E. coli also enhanced the antimicrobial activity when it was added to the medium in the form of living cells or as cell debris after cellular fractionation. No inducing activity was observed with addition of cell-free supernatant of E. coli cultures, suggesting that inducing factor is associated to the cells. Polyacrylamide gel electrophoresis revealed that additional peptide bands are secreted when B. amyloliquefaciens was cultivated in the presence of cell debris of E. coli. These results suggest that the presence of intact or inactivated E. coli enhanced the synthesis of antimicrobial peptides by B. amyloliquefaciens LBM 5006.

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

NARCIS (Netherlands)

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

2010-01-01

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

Patterns of Adherence of Helicobacter pylori Clinical Isolates to Epithelial Cells, and its Association with Disease and with Virulence Factors.

Science.gov (United States)

Vázquez-Jiménez, Flor Elizabeth; Torres, Javier; Flores-Luna, Lourdes; Cerezo, Silvia Giono; Camorlinga-Ponce, Margarita

2016-02-01

Adherence to the gastric epithelium is one of the most important steps of Helicobacter pylori to remain and cause disease. The aim of this study was to analyze whether H. pylori isolates from patients with different gastroduodenal diseases present differences in the pattern of adherence to gastric epithelial cells (AGS), in the ability to induce IL-8, and in the presence of virulence genes. We tested 75 H. pylori strains isolated from nonatrophic gastritis, gastric cancer, and duodenal ulcer patients. The adhesion pattern and IL-8 induction were determined in AGS cells, and invasion of AGS cells was studied using a gentamicin protection assay. The IL-8 levels induced were determined by ELISA. Helicobacter pylori strains presented diffuse adherence (DA) and localized (LA) adherence patterns, similar to those described for enteropathogenic E. coli (EPEC), were observed in AGS cells. A DA pattern was observed in 57% and LA in 43% of the strains, and DA was more frequent in isolates from patients with gastric cancer (p = 0.044). Strains with a LA pattern induced higher levels of IL-8 (p = 0.042) in AGS cells. The adherence pattern was not associated with neither invasiveness nor with the presence of virulence genes. Our study shows that H. pylori strains present adherence patterns to AGS cells resembling those observed in EPEC and that these patterns may be associated with disease and with activity on AGS cells. © 2015 John Wiley & Sons Ltd.

Association of vitamin D status with incidence of enterotoxigenic, enteropathogenic and enteroaggregative Escherichia coli diarrhoea in children of urban Bangladesh.

Science.gov (United States)

Ahmed, A M S; Soares Magalhaes, R J; Long, K Z; Ahmed, T; Alam, Md A; Hossain, Md I; Islam, Md M; Mahfuz, M; Mondal, D; Haque, R; Mamun, A A

2016-08-01

To evaluate the association between vitamin D status and diarrhoeal episodes by enterotoxigenic (ETEC), enteropathogenic (EPEC) and enteroaggregative (EAEC) E. coli in underweight and normal-weight children aged 6-24 months in urban Bangladesh. Cohorts of 446 normal-weight and 466 underweight children were tested separately for ETEC, EPEC and EAEC from diarrhoeal stool samples collected during 5 months of follow-up while considering vitamin D status at enrolment as the exposure. Cox proportional hazards models with unordered failure events of the same type were used to determine diarrhoeal risk factors after adjusting for sociodemographic and concurrent micronutrient status. Vitamin D status was not independently associated with the risk of incidence of ETEC, EPEC and EAEC diarrhoea in underweight children, but moderate-to-severe retinol deficiency was associated with reduced risk for EPEC diarrhoea upon adjustment. Among normal-weight children, insufficient vitamin D status and moderate-to-severe retinol deficiency were independently associated with 44% and 38% reduced risk of incidence of EAEC diarrhoea, respectively. These children were at higher risk of ETEC diarrhoea with vitamin D deficiency status when adjusted for micronutrient status only. This study demonstrates for the first time that normal-weight children with insufficient vitamin D status have a reduced risk of EAEC diarrhoea than children with sufficient status. Moderate-to-severe deficiency of serum retinol is associated with reduced risk of EPEC and EAEC diarrhoea in underweight and normal-weight children. © 2016 John Wiley & Sons Ltd.

Etiology of Diarrhea in Young Children in Denmark: a Case-Control Study

Science.gov (United States)

Olesen, Bente; Neimann, Jacob; Böttiger, Blenda; Ethelberg, Steen; Schiellerup, Peter; Jensen, Charlotte; Helms, Morten; Scheutz, Flemming; Olsen, Katharina E. P.; Krogfelt, Karen; Petersen, Eskild; Mølbak, Kåre; Gerner-Smidt, Peter

2005-01-01

Infectious gastroenteritis is one of the most common diseases in young children. To clarify the infectious etiology of diarrhea in Danish children less than 5 years of age, we conducted a 2-year prospective case-control study. Stools from 424 children with diarrhea and 870 asymptomatic age-matched controls were examined, and their parents were interviewed concerning symptoms. Rotavirus, adenovirus, and astrovirus were detected by enzyme-linked immunosorbent assay, and norovirus and sapovirus were detected by PCR. Salmonella, thermotolerant Campylobacter, Yersinia, Shigella, and Vibrio spp. were detected by standard methods. Shiga toxin-producing (STEC), attaching-and-effacing (A/EEC), enteropathogenic (EPEC), enterotoxigenic, enteroinvasive, and enteroaggregative Escherichia coli were detected by using colony hybridization with virulence gene probes and serotyping. Parasites were detected by microscopy. Overall, a potential pathogen was found in 54% of cases. More cases than controls were infected with rotavirus, Salmonella, norovirus, adenovirus, Campylobacter, sapovirus, STEC, classical EPEC, Yersinia, and Cryptosporidium strains, whereas A/EEC, although common, was not associated with illness. The single most important cause of diarrhea was rotavirus, which points toward the need for a childhood vaccine for this pathogen, but norovirus, adenovirus, and sapovirus were also major etiologies. Salmonella sp. was the most common bacterial pathogen, followed by Campylobacter, STEC, Yersinia, and classical EPEC strains. A/EEC not belonging to the classical EPEC serotypes was not associated with diarrhea, underscoring the importance of serotyping for the definition of EPEC. PMID:16081890

Proliferation of Prostate Stromal Cell Induced by Benign Prostatic Hyperplasia Epithelial Cell Stimulated With Trichomonas vaginalis via Crosstalk With Mast Cell.

Science.gov (United States)

Kim, Jung-Hyun; Kim, Sang-Su; Han, Ik-Hwan; Sim, Seobo; Ahn, Myoung-Hee; Ryu, Jae-Sook

2016-11-01

Chronic inflammation has a role in the pathogenesis of benign prostatic hyperplasia (BPH) and prostate cancer. Mast cells have been detected in chronic inflammatory infiltrate of the prostate, and it is possible that the interaction between prostate epithelial cells and Trichomonas vaginalis influences the activity of mast cells in the prostate stroma. Activated mast cells might influence the biological functions of nearby tissues and cells. In this study, we investigated whether mast cells reacted with the culture supernatant of BPH epithelial cells infected with T. vaginalis may induce the proliferation of prostate stromal cells. To measure the proliferation of prostate stromal cells in response to chronic inflammation caused by the infection of BPH-1 cells with T. vaginalis, the CCK-8 assay and wound healing assay were used. ELISAs, quantitative real-time PCR, western blotting and immunofluorescence were used to measure the production and expression of inflammatory cytokine and cytokine receptor. BPH-1 cells incubated with live trichomonads produced increased levels of CCL2, IL-1β, IL-6, and CXCL8, and induced the migration of mast cells and monocytes. When the culture supernatant of BPH-1 cells stimulated with trichomonads (TCM) was added to mast cells, they became activated, as confirmed by release of β-hexosaminidase and CXCL8. Prostate stromal cells incubated with the culture supernatant of mast cells activated with TCM (M-TCM) proliferated and expressed increased levels of CXCL8, CCL2, and the cytokine receptors CXCR1 and CCR2. Blocking the chemokine receptors reduced the proliferation of stromal cells and also decreased the production of CXCL8 and CCL2. Moreover, the expression of FGF2, cyclin D1, and Bcl-2 was increased in the proliferated stromal cells stimulated with M-TCM. Additionally, the M-TCM-treated stromal cells were more invasive than control cells. The inflammatory mediators released by BPH epithelial cells in response to infection by

A mutation (radA100) in Escherichia coli that selectively sensitizes cells grown in rich medium to X- or U.V.-radiation, or methyl methanesulphonate

International Nuclear Information System (INIS)

Diver, W.P.; Sargentini, N.J.; Smith, K.C.

1982-01-01

The radA100 mutant was isolated from Escherichia coli K-12 after mutagenesis with N-methyl-N'-nitro-N-nitrosoguanidine and selection for gamma radiation sensitivity. The radA100 mutation sensitized stationary phase cells to X-rays if they had been grown in glucose-supplemented rich medium, but not if they had been grown in nonsupplemented rich medium (indicating a defect in glucose-induced resistance). Similarly, logarithmic phase cells were sensitized to X-rays, U.V. radiation and methyl methanesulphonate if they had been grown in rich medium, but not if they had been grown in minimal medium (indicating a defect in medium-dependent resistance). Relative to the wild-type strain, the radA100 mutant was deficient in the repair of X-ray-induced DNA single-strand breaks when grown to logarithmic phase in rich medium, but not when grown in minimal medium. This is a novel mutation amongst the known DNA repair defects in that it did not sensitize logarithmic phase cells, grown in minimal medium, to either X- or U.V.-radiation. (author)

Modeling and analysis of strategic forward contracting in transmission constrained power markets

International Nuclear Information System (INIS)

Yu, C.W.; Chung, T.S.; Zhang, S.H.; Wang, X.

2010-01-01

Taking the effects of transmission network into account, strategic forward contracting induced by the interaction of generation firms' strategies in the spot and forward markets is investigated. A two-stage game model is proposed to describe generation firms' strategic forward contracting and spot market competition. In the spot market, generation firms behave strategically by submitting bids at their nodes in a form of linear supply function (LSF) and there are arbitrageurs who buy and resell power at different nodes where price differences exceed the costs of transmission. The owner of the grid is assumed to ration limited transmission line capacity to maximize the value of the transmission services in the spot market. The Cournot-type competition is assumed for the strategic forward contract market. This two-stage model is formulated as an equilibrium problem with equilibrium constraints (EPEC); in which each firm's optimization problem in the forward market is a mathematical program with equilibrium constraints (MPEC) and parameter-dependent spot market equilibrium as the inner problem. A nonlinear complementarity method is employed to solve this EPEC model. (author)

Complement protein C1q induces maturation of human dendritic cells

DEFF Research Database (Denmark)

Csomor, Eszter; Bajtay, Zsuzsa; Sándor, Noémi

2007-01-01

Maturation of dendritic cells (DCs) is known to be induced by several stimuli, including microbial products, inflammatory cytokines and immobilized IgG, as demonstrated recently. Since immune complexes formed in vivo also contain C1q, moreover apoptotic cells and several pathogens fix C1q...... activity of the cells was assessed by measuring cytokine secretion and their ability to activate allogeneic T lymphocytes. Cytokine production by T cells co-cultured with C1q-matured DCs was also investigated. C1q, but not the structurally related mannose-binding lectin was found to bind to imMDC in a dose......-dependent manner and induced NF-kappaB translocation to the nucleus. Immobilized C1q induced maturation of MDCs and enhanced secretion of IL-12 and TNF-alpha, moreover, elevated their T-cell stimulating capacity. As IFN-gamma levels were increased in supernatants of MDC-T cell co-cultures, our data suggest that C1...

Fluid-Driven Deformation of a Soft Porous Medium

Science.gov (United States)

Lutz, Tyler; Wilen, Larry; Wettlaufer, John

2017-11-01

Viscous drag forces resisting the flow of fluid through a soft porous medium are maintained by restoring forces associated with deformations in the solid matrix. We describe experimental measurements of the deformation of foam under a pressure-driven flow of water along a single axis. Image analysis techniques allow tracking of the foam displacement while pressure sensors allow measurement of the fluid pressure. Experiments are performed for a series of different pressure heads ranging from 10 to 90 psi, and the results are compared to theory. This work builds on previous measurements of the fluid-induced deformation of a bed of soft hydrogel spheres. Compared to the hydrogel system, foams have the advantage that the constituents of the porous medium do not rearrange during an experiment, but they have the disadvantage of having a high friction coefficient with any boundaries. We detail strategies to characterize and mitigate the effects of friction on the observed foam deformations.

Carboxymethyl cellulase and cellobiase production by Clostridium acetobutylicum in an industrial fermentation medium.

OpenAIRE

Allcock, E R; Woods, D R

1981-01-01

The production of a carboxymethyl cellulase and a cellobiase by Clostridium acetobutylicum was demonstrated. In liquid medium the carboxymethyl cellulase was induced by molasses, and it was not repressed by glucose. Optimum carboxymethyl cellulase activity occurred at pH 4.6 and 37 degrees C.

Analysis of interleukin 1 mediated radioprotection

International Nuclear Information System (INIS)

Manori, I.; Kushilevsky, A.; Weinstein, Y.

1986-01-01

The potential value of interleukin 1(IL-1) containing supernatants as a radioprotective agent was evaluated. It was found that the response of irradiated thymocytes to mitogens was partially restored if IL-1 containing supernatants was included in the culture medium immediately after irradiation. A delay of 24 h in the addition of IL-1 and mitogen abrogated the radioprotection effect. Under the same conditions IL-2 containing supernatants were effective, suggesting that the dose modifying effect of IL-1 acts through induction of IL-2 elaboration. The results of the present study may be important in cases where it is necessary to restore depressed immune response resulting from irradiation accidents or radiotherapy. (author)

A medium energy neutron deep penetration experiment

International Nuclear Information System (INIS)

Amian, W.; Cloth, P.; Druecke, V.; Filges, D.; Paul, N.; Schaal, H.

1986-11-01

A deep penetration experiment conducted at the Los Alamos WNR facility's Spallation Neutron Target is compared with calculations using intra-nuclear-cascade and S N -transport codes installed at KFA-IRE. In the experiment medium energy reactions induced by neutrons between 15 MeV and about 150 MeV inside a quasi infinite slab of iron have been measured using copper foil monitors. Details of the experimental procedure and the theoretical methods are described. A comparison of absolute reaction rates for both experimentally and theoretically derived reactions is given. The present knowledge of the corresponding monitor reaction cross sections is discussed. (orig.)

Photon induced reactions

International Nuclear Information System (INIS)

Mecking, B.A.

1982-04-01

Various aspects of medium energy nuclear reactions induced by real photons are reviewed. Special emphasis is put on high accuracy experiments that will become possible with the next generation of electron accelerators. (orig.)

Using induced pluripotent stem cell-derived conditional medium to attenuate the light-induced photodamaged retina of rats

Directory of Open Access Journals (Sweden)

Hua-Ming Chang

2015-03-01

Conclusion: The conditional medium of iPSCs contains plenty of cytoprotective, immune-modulative and rescue chemicals, contributing to the maintenance of neuronal function and retinal layers in light-damaged retina compared with apoptotic iPSC-CM and PBS. The antiapoptotic effect of iPSC-CM also shows promise in restoring damaged neurons. This result demonstrates that iPSC-CM may serve as an alternative to cell therapy alone to treat retinal light damage and maintain functional and structural integrity of the retina.

Microwave activation of palladium nanoparticles for enhanced ethanol electrocatalytic oxidation reaction in alkaline medium

CSIR Research Space (South Africa)

Rohwer, MB

2015-02-01

Full Text Available in alkaline medium (in terms of high mass activity stability and fast reaction kinetics). The remarkable microwave-induced properties on the Pd catalyst promise to revolutionize the use of microwave for catalyst activation for enhanced heterogeneous catalysis...

Antagonistic activities of some Bifidobacterium sp. strains isolated from resident infant gastrointestinal microbiota on Gram-negative enteric pathogens.

Science.gov (United States)

Delcaru, Cristina; Alexandru, Ionela; Podgoreanu, Paulina; Cristea, Violeta Corina; Bleotu, Coralia; Chifiriuc, Mariana Carmen; Bezirtzoglou, Eugenia; Lazar, Veronica

2016-06-01

The gastrointestinal microbiota contributes to the consolidation of the anti-infectious barrier against enteric pathogens. The purpose of this study was to investigate the influence of Bifidobacterium sp. strains, recently isolated from infant gastrointestinal microbiota on the in vitro growth and virulence features expression of enteropathogenic bacterial strains. The antibacterial activity of twelve Bifidobacterium sp. strains isolated from human feces was examined in vitro against a wide range of Gram negative pathogenic strains isolated from 30 infant patients (3 days to 5 years old) with diarrhea. Both potential probiotic strains (Bifidobacterium longum, Bifidobacterium pseudocatenulatum, Bifidobacterium catenulatum, Bifidobacterium breve, Bifidobacterium ruminantium) and enteropathogenic strains (EPEC, EIEC, Klebsiella pneumoniae, Salmonella sp., Yersinia enterocolitica, Pseudomonas aeruginosa) were identified by MALDI-TOF and confirmed serologically when needed. The bactericidal activity, growth curve, adherence to the cellular HEp-2 substratum and production of soluble virulence factors have been assessed in the presence of different Bifidobacterium sp. cultures and fractions (whole culture and free-cell supernatants). Among the twelve Bifidobacterium sp. strains, the largest spectrum of antimicrobial activity against 9 of the 18 enteropathogenic strains was revealed for a B. breve strain recently isolated from infant intestinal feces. The whole culture and free-cell supernatant of B. breve culture decreased the multiplication rate, shortened the log phase and the total duration of the growth curve, with an earlier entrance in the decline phase and inhibited the adherence capacity to a cellular substratum and the swimming/swarming motility too. These results indicate the significant probiotic potential of the B. breve strain. Copyright © 2016 Elsevier Ltd. All rights reserved.

Hypoxia-inducible transcription factor-1α promotes hypoxia-induced A549 apoptosis via a mechanism that involves the glycolysis pathway

International Nuclear Information System (INIS)

Luo, FengMing; Liu, XiaoJing; Yan, NaiHong; Li, ShuangQing; Cao, GuiQun; Cheng, QingYing; Xia, QingJie; Wang, HongJing

2006-01-01

Hypoxia-inducible transcription factor-1α (HIF-1α), which plays an important role in controlling the hypoxia-induced glycolysis pathway, is a 'master' gene in the tissue hypoxia response during tumor development. However, its role in the apoptosis of non-small cell lung cancer remains unknown. Here, we have studied the effects of HIF-1α on apoptosis by modulating HIF-1α gene expression in A549 cells through both siRNA knock-down and over-expression. A549 cells were transfected with a HIF-1α siRNA plasmid or a HIF-1α expression vector. Transfected cells were exposed to a normoxic or hypoxic environment in the presence or absence of 25 mM HEPES and 2-deoxyglucose (2-DG) (5 mM). The expression of three key genes of the glycolysis pathway, glucose transporter type 1(GLUT1), phosphoglycerate kinase 1(PGK1), and hexokinase 1(HK1), were measured using real-time RT-PCR. Glycolysis was monitored by measuring changes of pH and lactate concentration in the culture medium. Apoptosis was detected by TUNEL assay and flow cytometry. Knocking down expression of HIF-1α inhibited the glycolysis pathway, increased the pH of the culture medium, and protected the cells from hypoxia-induced apoptosis. In contrast, over-expression of HIF-1α accelerated glycolysis in A549 cells, decreased the pH of the culture medium, and enhanced hypoxia-induced apoptosis. These effects of HIF-1α on glycolysis, pH of the medium, and apoptosis were reversed by treatment with the glycolytic inhibitor, 2-DG. Apoptosis induced by HIF-1α over-expression was partially inhibited by increasing the buffering capacity of the culture medium by adding HEPES. During hypoxia in A549 cells, HIF-1α promotes activity of the glycolysis pathway and decreases the pH of the culture medium, resulting in increased cellular apoptosis

Contribution of radiation-induced, nitric oxide-mediated bystander effect to radiation-induced adaptive response.

Science.gov (United States)

Matsumoto, H.; Ohnishi, T.

There has been a recent upsurge of interest in radiation-induced adaptive response and bystander effect which are specific modes in stress response to low-dose low-dose rate radiation Recently we found that the accumulation of inducible nitric oxide NO synthase iNOS in wt p53 cells was induced by chronic irradiation with gamma rays followed by acute irradiation with X-rays but not by each one resulting in an increase in nitrite concentrations of medium It is suggested that the accumulation of iNOS may be due to the depression of acute irradiation-induced p53 functions by pre-chronic irradiation In addition we found that the radiosensitivity of wt p53 cells against acute irradiation with X-rays was reduced after chronic irradiation with gamma rays This reduction of radiosensitivity of wt p53 cells was nearly completely suppressed by the addition of NO scavenger carboxy-PTIO to the medium This reduction of radiosensitivity of wt p53 cells is just radiation-induced adaptive response suggesting that NO-mediated bystander effect may considerably contribute to adaptive response induced by radiation

Effect of amino acids on tannase biosynthesis by Bacillus licheniformis KBR6.

Science.gov (United States)

Mohapatra, Pradeep K Das; Pati, Bikas R; Mondal, Keshab C

2009-04-01

Microbial tannase (tannin acyl hydrolase, EC 3.1.1.20), a hydrolysable tannin-degrading enzyme, has gained importance in various industrial processes, and is used extensively in the manufacture of instant tea, beer, wine, and gallic acid. Tannase is an inducible enzyme, and hydrolysable tannin, especially tannic acid, is the sole inducer. This study is of the effect of various amino acids and their analogues on tannase biosynthesis by Bacillus licheniformis KBR6 to ascertain the mode of action of these growth factors on tannase biosynthesis from microbial origin. Enzyme production was carried out in enriched tannic acid medium through submerged fermentation for 20 h at 35 degrees C. Different amino acids at a concentration of 0.05 g% (w/v) were added to the culture medium immediately after sterilization. Culture supernatant was used as the source of the enzyme and the quantity of tannase was estimated by the colorimetric assay method. Growth of the organism was estimated according to biomass dry weight. Maximum tannase (2.87-fold that of the control) was synthesized by B. licheniformis KBR6 when alanine was added to the culture medium. Other amino acids, such as DL-serine, L-cystine, glycine, L-ornithine, aspartic acid, L-glutamic acid, DL-valine, L-leucine and L-lysine, also induced tannase synthesis. L-Cysteine monohydrochloride and DL-threonine were the most potent inhibitors. Regulation of tannase biosynthesis by B. licheniformis in the presence of various amino acids is shown. This information will be helpful for formulating an enriched culture medium for industrial-scale tannase production.

STEREOCHEMICAL ASPECTS OF HYDRATION OF CARBOHYDRATES IN AQUEOUS-SOLUTIONS .2. KINETIC MEDIUM EFFECTS

NARCIS (Netherlands)

GALEMA, SA; BLANDAMER, MJ; ENGBERTS, JBFN

1992-01-01

Rate constants for the hydrolysis of 1-benzoyl-3-phenyl-1,2,4-triazole in aqueous solutions of carbohydrates have been measured as a function of molality and nature of added mono- and disaccharides. The kinetic medium effects induced by the carbohydrates originate from hydration sphere overlap

Increased hydrazine during partial nitritation process in upflow air-lift reactor fed with supernatant of anaerobic digester effluent

Energy Technology Data Exchange (ETDEWEB)

Choi, Jeongdong [University of Alberta, Alberta (Canada); Jung, Sokhee [Samsung SDS, Seoul (Korea, Republic of); Ahn, Young-Ho [Yeungnam University, Gyungsan (Korea, Republic of)

2013-06-15

The optimal balance of ammonium and nitrite is essential for successful operation of the subsequent anammox process. We conducted a partial nitritation experiment using an upflow air-lift reactor to provide operational parameters for achieving the optimal ratio of ammonium to nitrite, by feeding supernatant of anaerobic digester effluent, high-nitrogen containing rejection water. Semi-continuous operation results show that HRT should be set between 15 and 17 hours to achieve the optimum ration of 1.3 of NO{sub 2}-N/NH{sub 4}-N. In the UAR, nitritation was the dominant reaction due to high concentration of ammonia and low biodegradable organics. The influent contained low concentrations of hydroxylamine and hydrazine. However, hydrazine increased during partial nitritation by ⁓60-130% although there was no potential anammox activity in the reactor. The partial nitritation process successfully provided the ratio of nitrogen species for the anammox reaction, and relived the nitrite restraint on the anammox activity by increasing hydrazine concentration.

Inflammation promotes oral squamous carcinoma immune evasion via induced programmed death ligand-1 surface expression.

Science.gov (United States)

Lu, Wanlu; Lu, Libing; Feng, Yun; Chen, Jiao; Li, Yan; Kong, Xiangli; Chen, Sixiu; Li, Xiaoyu; Chen, Qianming; Zhang, Ping

2013-05-01

The association between inflammation and cancer provides a new target for tumor biotherapy. The inflammatory cells and molecules within the tumor microenvironment have decisive dual roles in antitumor immunity and immune evasion. In the present study, phytohemagglutinin (PHA) was used to stimulate peripheral blood mononuclear cells (PBMCs) to simulate the tumor inflammatory microenvironment. The effect of immune cells and inflammatory cytokines on the surface expression of programmed cell death-1 ligand 1 (PD-L1) and tumor immune evasion was investigated using flow cytometry (FCM) and an in vivo xenotransplantation model. Based on the data, PHA-activated, but not resting, immune cells were able to promote the surface expression of PD-L1 in Tca8113 oral squamous carcinoma cells via the secretion of inflammatory cytokines, but not by cell-cell contact. The majority of the inflammatory cytokines had no significant effect on the proliferation, cell cycle progression and apoptosis of the Tca8113 cells, although they each induced the expression of PD-L1 in a dose-dependent manner. In total, 99% of the Tca8113 cells expressed PD-L1 following treatment with the supernatant of PHA-stimulated PBMCs. The PHA-supernatant pretreated Tca8113 cells unusually induced Tca8113 antigen-specific CD8 + T cell apoptosis in vitro and the evasion of antigen-specific T cell attraction in a nude mouse tumor-bearing model. These results indicate a new mechanism for the promotion of tumor immune evasion by the tumor inflammatory microenvironment.

Medium-induced change of the optical response of metal clusters in rare-gas matrices

Science.gov (United States)

Xuan, Fengyuan; Guet, Claude

2017-10-01

Interaction with the surrounding medium modifies the optical response of embedded metal clusters. For clusters from about ten to a few hundreds of silver atoms, embedded in rare-gas matrices, we study the environment effect within the matrix random phase approximation with exact exchange (RPAE) quantum approach, which has proved successful for free silver clusters. The polarizable surrounding medium screens the residual two-body RPAE interaction, adds a polarization term to the one-body potential, and shifts the vacuum energy of the active delocalized valence electrons. Within this model, we calculate the dipole oscillator strength distribution for Ag clusters embedded in helium droplets, neon, argon, krypton, and xenon matrices. The main contribution to the dipole surface plasmon red shift originates from the rare-gas polarization screening of the two-body interaction. The large size limit of the dipole surface plasmon agrees well with the classical prediction.

MicroRNA Expression in Salivary Supernatant of Patients with Pancreatic Cancer and Its Relationship with ZHENG

Directory of Open Access Journals (Sweden)

Song Gao

2014-01-01

Full Text Available In traditional Chinese medicine (TCM, diagnosis and prescriptions are based on the signs and symptoms which are recognized as ZHENG. The cornerstone of TCM is to differentially treat one ZHENG from others, which is also known as syndrome differentiation, and this relies on the gathering of clinical information through inspection, auscultation and olfaction, inquiry, and palpation. However, the biomolecular basis of the ZHENG remains unclear. In this study, the expressions of 384 cancer-related miRNAs in salivary supernatant of patients with pancreatic cancer were assessed by miRNA polymerase chain reaction (PCR array, and the different expression patterns of miRNA in three different groups of ZHENG were studied with use of real-time quantitative PCR (qRT-PCR. Some miRNAs were found to be specifically expressed in some ZHENGs, for instance, miR-17, miR-21, and miR-181b in Shi-Re ZHENG and miR-196a in Pi-Xu ZHENG. This indicates that these miRNAs may play important roles in different ZHENG condition. Therefore, this study to some extent revealed the molecular basis of TCM ZHENG in pancreatic cancer.

Bench-scale cross flow filtration of Tank S-107 sludge slurries and Tank C-107 supernatant

International Nuclear Information System (INIS)

Geeting, J.G.H.; Reynolds, B.A.

1996-10-01

Hanford tank waste filtration experiments were conducted using a bench-scale cross flow filter on 8 wt%, 1.5 wt%, and 0.05 wt% Tank S- 107 sludge slurries and on Tank C-107 supernatant. For comparison, two simulants each with solids loadings of 8 wt% and 0.05 wt% were also tested. The purpose of the tests was to determine the efficacy of cross flow filtration on slurries of various solids loadings. -In addition, filtrate flux dependency on axial velocity and transmembrane pressure was sought so that conditions for future experiments might be better selected. The data gathered are compared to the simulants and three cross flow filtration models. A two- parameter central composite design which tested. transmembrane pressure from 5 to 40 psig and axial Velocity from 3 to 9 ft/s was used for all feeds. The cross flow filter effectively removed solids from the liquid, as 19 of 20 filtrate samples had particle concentrations below the resolution limit of the photon correlation spectrometer used in the Hanford Radiocolloid Laboratory. Radiochemical analysis indicate that all filtrate samples were below Class A waste classification standards for 9OSr and transuranics

Determining antioxidant activities of lactobacilli cell-free supernatants by cellular antioxidant assay: a comparison with traditional methods.

Directory of Open Access Journals (Sweden)

Jiali Xing

Full Text Available Antioxidant activity of lactic acid bacteria is associated with multiple health-protective effects. Traditional indexes of chemical antioxidant activities poorly reflect the antioxidant effects of these bacteria in vivo. Cellular antioxidant activity (CAA assay was used in this study to determine the antioxidant activity of cell-free supernatants (CFSs of 10 Lactobacillus strains. The performance of the CAA assay was compared with that of four chemical antioxidant activity assays, namely, DPPH radical scavenging, hydroxyl radical scavenging (HRS, reducing power (RP, and inhibition of linoleic acid peroxidation (ILAP. Results of the CAA assay were associated with those of DPPH and ILAP assays, but not with those of RP and HRS assays. The inter- and intra-specific antioxidant activities of CFS were characterized by chemical and CAA assays. L. rhamnosus CCFM 1107 displayed a high antioxidative effect similar to positive control L. rhamnosus GG ATCC 53103 in all of the assays. The CAA assay is a potential method for the detection of antioxidant activities of lactobacilli CFSs.

Tryptase potentiates enteric nerve activation by histamine and serotonin: Relevance for the effects of mucosal biopsy supernatants from irritable bowel syndrome patients.

Science.gov (United States)

Ostertag, D; Annahazi, A; Krueger, D; Michel, K; Demir, I E; Ceyhan, G O; Zeller, F; Schemann, M

2017-09-01

We previously showed that mucosal biopsy supernatants from irritable bowel syndrome patients activated neurons despite low concentrations of tryptase, histamine, and serotonin which individually would not cause spike discharge. We studied the potentiating responses between these mediators on excitability of enteric neurons. Calcium-imaging was performed using the calcium-sensitive dye Fluo-4 AM in human submucous plexus preparations from 45 individuals. Histamine, serotonin, and tryptase were applied alone and in combinations to evaluate nerve activation which was assessed by analyzing increase in intracellular Ca 2+ ([Ca 2+ ] i ), the proportion of responding neurons and the product of both defined as Ca-neuroindex (NI). Protease activated receptor (PAR) 2 activating peptide, PAR2 antagonist and the serine protease-inhibitor FUT-175 were used to particularly investigate the role of proteases. Histamine or serotonin (1 μmol/L each) evoked only few small responses (median NI [25%/75%]: 0 [0/148]; 85 [0/705] respectively). Their combined application evoked statistically similar responses (216 [21/651]). Addition of the PAR2 activator tryptase induced a significantly higher Ca-NI (1401 [867/4075]) compared to individual application of tryptase or to coapplied histamine and serotonin. This synergistic potentiation was neither mimicked by PAR2 activating peptide nor reversed by the PAR2 antagonist GB83, but abolished by FUT-175. We observed synergistic potentiation between histamine, serotonin, and tryptase in enteric neurons, which is mediated by proteolytic activity rather than PAR2 activation. This explained neuronal activation by a cocktail of these mediators despite their low concentrations and despite a relatively small PAR2-mediated response in human submucous neurons. © 2017 John Wiley & Sons Ltd.

Optical bistability induced by quantum coherence in a negative index atomic medium

International Nuclear Information System (INIS)

Zhang Hong-Jun; Sun Hui; Li Jin-Ping; Yin Bao-Yin; Guo Hong-Ju

2013-01-01

Bistability behaviors in an optical ring cavity filled with a dense V-type four-level atomic medium are theoretically investigated. It is found that the optical bistability can appear in the negative refraction frequency band, while both the bistability and multi-stability can occur in the positive refraction frequency bands. Therefore, optical bistability can be realized from conventional material to negative index material due to quantum coherence in our scheme. (electromagnetism, optics, acoustics, heat transfer, classical mechanics, and fluid dynamics)

Caffeine and D sub 2 O medium interact in affecting the expression of radiation-induced potentially lethal damage

Energy Technology Data Exchange (ETDEWEB)

Utsumi, H. (Kyoto Univ. (Japan). Radiation Biology Center); Elkind, M.M. (Colorado State Univ., Fort Collins, CO (United States). Dept. of Radiological Health Sciences)

1991-10-01

Earlier work has been extended to compare the killing of long-phase V79 Chinese hamster cells by ionizing radiation when they are treated immediately after irradiation with medium containing either caffeine or 90% D{sub 2}O. The object was to determine if the enhanced killing due to post-treatment with caffeine, or D{sub 2}O, resulted from action on the same sector of potentially lethal damage as appeared to be the case for hypertonic shock and D{sub 2}O medium. The treatments by themselves were not toxic to unirradiated cells. We found that the enhanced expression of potentially lethal damage by post-treatment with caffeine or D{sub 2}O medium is similar. For example, the kinetic of the repair of the potentially lethal damage expressible by either post-treatment was similar, and an additive enhancement of potentially lethal damage occurred when the two treatments were administered sequentially. These findings suggest that caffeine and D{sub 2}O medium affect the same sector of potentially lethal damage. When the two treatments were combined, however, they competed with each other. Thus, although caffeine and D{sub 2}O medium act on the same sector of potentially lethal damage they do so differently, suggesting that more than one pathway of the expression of radiation damage can result in the same phenotypic effect. (author).

Boildown Study on Supernatant Liquid Retrieved from AP-107 in May 2010

Energy Technology Data Exchange (ETDEWEB)

Callaway, W. S. [Washington River Protection Solutions, LLC, Richland, WA (United States); Page, J. S. [Washington River Protection Solutions, LLC, Richland, WA (United States)

2013-02-12

A boildown study was completed on a composite prepared from supernatant liquid grab samples retrieved from tank 241-AP-107 in May of 2010. The composite was a clear, yellow liquid containing no visible solids at hot cell ambient temperatures (25-27 °C). The density of the test composite was 1.216 g/mL at 26.8 °C. The boiling temperature curves generated at three reduced pressures—40-, 60-, and 80 Torr—displayed steadily increasing boiling temperatures with increasing volume reduction with no significant discontinuities. Only minimal foaming was observed after the volume reduction proceeded beyond 50 %WVR (percent waste volume reduction). The bulk densities (D{sub Bulk}{sup 18 °C}) and quantities of settled and centrifuged solids present were measured on samples of the boildown concentrates that were kept at 18 °C for 7-8 days. Estimated values of the bulk densities of the concentrates at 60-Torr boiling temperatures (D{sub Bulk}{sup 60 Torr}) were also calculated. Solids were observed in all boildown concentrates at process temperatures, at hot cell ambient temperatures (25-27 °C), and at 18 °C. The quantity of solids found in the cooled concentrates increased slowly through 50.2 %WVR. The quantity of solids found in concentrates after 54.0 %WVR was noticeably greater. Beyond 54.0 %WVR, the quantity of solids found in cooled concentrates increased dramatically. Analysis of boildown test samples indicated that sodium oxalate and sodium carbonate solids form in cooled concentrates after volume reduction of 8.4 %WVR or less. The major contributors to the large increase in the quantity of solids found in concentrates after 54 %WVR were sodium nitrate and sodium carbonate.

Spironolactone induces apoptosis in human mononuclear cells. Association between apoptosis and cytokine suppression

DEFF Research Database (Denmark)

Mikkelsen, Martin; Sønder, S U; Nersting, J

2006-01-01

Spironolactone (SPIR) has been described to suppress accumulation of pro-inflammatory cytokines. Here, the suppression of TNF-alpha in lipopolysaccharide (LPS)-stimulated mononuclear cell cultures was confirmed. However, SPIR was also found to induce apoptosis, prompting the investigations...... of a possible association between the two effects: The apoptosis-inducing and the cytokine-suppressive effects of SPIR correlated with regard to the effective concentration range. Also, pre-incubation experiments demonstrated a temporal separation of the two effects of ... preceding apoptosis. An association between the two effects was also seen when testing several SPIR analogues. Contrary to TNF-alpha, the levels of IL-1beta increased in SPIR-treated cultures. However, the amount of IL-1beta in the supernatants depended upon the order of SPIR and LPS addition, as IL-1beta...

Genetic characterization of antibiotic resistance in enteropathogenic Escherichia coli carrying extended-spectrum beta-lactamases recovered from diarrhoeic rabbits.

Science.gov (United States)

Poeta, P; Radhouani, H; Gonçalves, A; Figueiredo, N; Carvalho, C; Rodrigues, J; Igrejas, G

2010-05-01

A total of 52 Escherichia coli strains isolated from diarrhoeic rabbits were investigated for their enteropathogenic E. coli (EPEC) pathotype by PCR amplification of eae and bfp virulence genes. A total of 22 EPEC isolates were identified, serotyped and studied for antibiotic resistance and screened for the detection of extended-spectrum beta-lactamases (ESBLs). The EPEC isolates belonged to three serogroups (O26, O92 and O103). The most common serogroup (O103:K-:H2) was observed among 17 EPEC strains, the O92:K-serogroup in three isolates (the antibiotic sensitive ones) and the remaining O26:K-serogroup in two isolates (the ESBLs isolates). Resistances to ampicillin and tetracycline were the most frequent and detected followed by resistance to nalidixic acid, streptomycin, trimethoprim-sulphamethoxazole, cefoxitin, gentamicin and ciprofloxacin. All the isolates were sensitive for amikacin, ceftazidime, aztreonam, imipenem, chloramphenicol, tobramycin and amoxicillin + clavulanic acid. Two isolates recovered from two adult animals showed an intermediate susceptibility to cefotaxime, and a positive screening test for ESBL was demonstrated in both. The bla(TEM) gene was demonstrated in the majority of ampicillin-resistant isolates. The aac(3)-II or aac(3)-IV genes were detected in the four gentamicin-resistant isolates. In addition, the aadA gene was detected in 60% of streptomycin-resistant isolates. The tet(A) or tet(B) genes were identified in all tetracycline-resistant isolates. A total of nine EPEC isolates showed the phenotype SXT-resistant, and the sul1 and/or sul2 and/or sul3 genes were detected in all of them. Our findings showed that the molecular detection by the eae and bfp genes by PCR followed by serotyping is useful for monitoring trends in EPEC infections of rabbits allowing the identification of their possible reservoirs. The detection of genes involved in the resistance to antibiotics of different families in a relatively high proportion of faecal E

Evaluation at the medium energy region for Pb-208 and Bi-209

International Nuclear Information System (INIS)

Fukahori, Tokio; Pearlstein, S.

1991-01-01

Medium energy nuclear data in the 1--1000 MeV range is necessary to accelerator applications which include spallation neutron sources for radioactive waste treatment and accelerator shielding design, medical applications which include isotopes production and radiation therapy, and space applications. For the design of fission and fusion reactors, the nuclear data file for neutrons below 20 MeV is available and well evaluated. Evaluated nuclear data for protons and data in the medium energy region, however, have not been prepared completely. Evaluation in the medium energy region was performed using the theoretical calculation code ALICE-P or experimental data. In this paper, the evaluation of neutron and proton induced nuclear data for Pb-208 and Bi-209 has been performed using ALICE-P, empirical calculations and new systematics for the fission cross section. The evaluated data are compiled for possible inclusion in the ENDF/B-VI High Energy File. 204 refs., 51 figs., 9 tabs

Radioadaptive response to the medium-mediated bystander induction of DNA strand breaks in HeLa cells

International Nuclear Information System (INIS)

Ikushima, T.; Okuyama, M.

2003-01-01

Full text: Numerous investigators have reported two cellular responses of importance at low doses that have a potential impact on the risk estimation of ionizing radiation. The radioadaptive response confers resistance to a subsequent dose by a low priming dose, while the bystander effect exaggerates the effect of small doses. The present study was conducted to examine the interaction of the radioadaptive response with the bystander effect in HeLa cells. The culture was irradiated with 0.5 to 8 Gy of 140 kVp X-rays and one hour later, the medium was taken, passed through a filter and transferred to the parallel culture of non-irradiated HeLa cells as non-targeted cells. After incubation for 30 min, the induced DNA damage was analyzed by the single cell gel-electrophoresis assay under alkaline or neutral conditions. The treatments resulted in a dose-dependent increase in tail moment under either conditions, indicating the induction of DNA single- and double-strand breaks. The clonogenic survival of non-irradiated cells was also reduced after they were cultured in the medium that was taken from irradiated cultures. Any change was not observed when the medium alone was irradiated. These results give the disputed evidence that certain genotoxic factor(s) released from irradiated cells into the culture medium can induce DNA strand breaks leading to cell death. It is also suggested that physical contact between irradiated and non-irradiated cells may not be required for the bystander effect. In adapted cells that were pre-exposed to 5 cGy of X-rays and cultured for 4 h beforehand, the yield of DNA strand breaks induced by X-rayed medium was reduced by about 50 %. The results, in conjunction with our early finding (Ikushima et al., 1996) suggest that the radioadaptive response resulting from such a low dose may diminish the bystander effect through an enhanced DNA repair function

PERFIL DE SENSIBILIDADE MICROBIANA IN VITRO DE LINHAGENS PATOGÊNICAS DE Escherichia coli ISOLADAS DE CARNE BOVINA

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Samira Pirola Santos Mantilla

2012-06-01

Full Text Available This study aimed to analyze the antimicrobial resistance of Escherichia coli strains (EPEC A, EPEC B, EPEC C, EIEC A e EIEC B isolated from bovine meat. The antimicrobial susceptibility test was evaluated using the technique described by the National Committee for Clinical Laboratory Standards. The strains were resistant to most antibiotics tested, and gentamicin showed the best efficiency, with 84.9% of the strains showing sensitivity. In addition, cefoxitin was the least effective antimicrobial agent, have a higher percentage of resistant strains. The multidrug resistance to antimicrobials is a finding of great importance to public health, as resistant pathogens may be conveyed to consumers by the ingestion of animal products, making difficult the treatment of bacterial infections and increasing the occurrence of bacteria resistant to antibiotics.

Q-PYTHIA: a medium-modified implementation of final state radiation

Science.gov (United States)

Armesto, Néstor; Cunqueiro, Leticia; Salgado, Carlos A.

2009-10-01

We present a Monte Carlo implementation, within PYTHIA, of medium-induced gluon radiation in the final state branching process. Medium effects are introduced through an additive term in the splitting functions computed in the multiple-soft scattering approximation. The observable effects of this modification are studied for different quantities as fragmentation functions and the hump-backed plateau, and transverse momentum and angular distributions. The anticipated increase of intra-jet multiplicities, energy loss of the leading particle and jet broadening are observed as well as modifications of naive expectations based solely on analytical calculations. This shows the adequacy of a Monte Carlo simulator for jet analyses. Effects of hadronization are found to wash out medium effects in the soft region, while the main features remain. To show the performance of the implementation and the feasibility of our approach in realistic experimental situations we provide some examples: fragmentation functions, nuclear suppression factors, jet shapes and jet multiplicities. The package containing the modified routines is available for public use. This code, which is not an official PYTHIA release, is called Q-PYTHIA. We also include a short manual to perform the simulations of jet quenching.

Method and apparatus for induced nuclear beta decay

International Nuclear Information System (INIS)

Reiss, H.

1986-01-01

This invention relates to a method and apparatus for inducing beta decay transition that are normally inhibited by angular momentum or parity considerations. According to one aspect of this invention a method of inducing nuclear beta decay transition comprises providing a medium which includes atomic nuclei that have forbidden beta decay transition in which the initial and final nuclear states do not have the same intrinsic parity or have total angular momenta which differ by more than one quantum unit of angular momentum, and applying to the medium an electromagnetic field which has an intensity sufficient to provide the angular momentum or intrinsic parity necessary to overcome the forbiddenness of the beta decay transition of the atomic nuclei, thereby to induce the beta decay transitions. According to another aspect of this invention an apparatus for inducing beta decay transition comprises a medium which includes atomic nuclei that have forbidden beta decay transitions in which the initial and final nuclear states do not have the same intrinsic parity or have total angular momenta which differ by more than one quantum unit of angular momentum, field producing means for producing an electromagnetic field in the medium and means for energising the field producing means to establish the field at an intensity sufficient to provide the angular momentum or intrinsic parity necessary to overcome the forbiddenness of the beta decay transitions of the atomic nuclei. The energy released in these induced nuclear transition is useful for the controlled production of power. The induced beta dacay transitions are also useful to reduce the halflives of long-lived fission product wastes from conventional nuclear fission power plants

Release of Periplasmic Nucleotidase Induced by Human Antimicrobial Peptide in E. coli Causes Accumulation of the Immunomodulator Adenosine.

Directory of Open Access Journals (Sweden)

Andreia Bergamo Estrela

Full Text Available Previous work by our group described that human β-defensin-2 induces accumulation of extracellular adenosine (Ado in E. coli cultures through a non-lytic mechanism causing severe plasmolysis. Here, we investigate the presence of AMP as a direct precursor and the involvement of a bacterial enzyme in the generation of extracellular Ado by treated bacteria. Following hBD-2 treatment, metabolites were quantified in the supernatants using targeted HPLC-MS/MS analysis. Microbial growth was monitored by optical density and cell viability was determined by colony forming units counts. Phosphatase activity was measured using chromogenic substrate pNPP. The results demonstrate that defensin-treated E. coli strain W releases AMP in the extracellular space, where it is converted to Ado by a bacterial soluble factor. An increase in phosphatase activity in the supernatant was observed after peptide treatment, similar to the effect of sucrose-induced osmotic stress, suggesting that the periplasmic 5'nucleotidase (5'-NT is released following the plasmolysis event triggered by the peptide. Ado accumulation was enhanced in the presence of Co2+ ion and inhibited by EDTA, further supporting the involvement of a metallo-phosphatase such as 5'-NT in extracellular AMP conversion into Ado. The comparative analysis of hBD-induced Ado accumulation in different E. coli strains and in Pseudomonas aeruginosa revealed that the response is not correlated to the peptide's effect on cell viability, but indicates it might be dependent on the subcellular distribution of the nucleotidase. Taken together, these data shed light on a yet undescribed mechanism of host-microbial interaction: a human antimicrobial peptide inducing selective release of a bacterial enzyme (E. coli 5'-NT, leading to the formation of a potent immunomodulator metabolite (Ado.

The innate defense antimicrobial peptides hBD3 and RNase7 are induced in human umbilical vein endothelial cells by classical inflammatory cytokines but not Th17 cytokines.

Science.gov (United States)

Burgey, Christine; Kern, Winfried V; Römer, Winfried; Sakinc, Türkan; Rieg, Siegbert

2015-05-01

Antimicrobial peptides are multifunctional effector molecules of innate immunity. In this study we investigated whether endothelial cells actively contribute to innate defense mechanisms by expression of antimicrobial peptides. We therefore stimulated human umbilical vein endothelial cells (HUVEC) with inflammatory cytokines, Th17 cytokines, heat-inactivated bacteria, bacterial conditioned medium (BCM) of Staphylococcus aureus and Streptococcus sanguinis, and lipoteichoic acid (LTA). Stimulation with single cytokines induced discrete expression of human β-defensin 3 (hBD3) by IFN-γ or IL-1β and of ribonuclease 7 (RNase7) by TNF-α without any effects on LL-37 gene expression. Stronger hBD3 and RNase7 induction was observed after combined stimulation with IL-1β, TNF-α and IFN-γ and was confirmed by high hBD3 and RNase7 peptide levels in cell culture supernatants. In contrast, Th17 cytokines or stimulation with LTA did not result in AMP production. Moreover, only BCM of an invasive S. aureus bacteremia isolate induced hBD3 in HUVEC. We conclude that endothelial cells actively contribute to prevent dissemination of pathogens at the blood-tissue-barrier by production of AMPs that exhibit microbicidal and immunomodulatory functions. Further investigations should focus on tissue-specific AMP induction in different endothelial cell types, on pathogen-specific induction patterns and potentially involved pattern-recognition receptors of endothelial cells. Copyright © 2015 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

Spiral–pacemaker interactions in a mathematical model of excitable medium

International Nuclear Information System (INIS)

Shajahan, T K; Borek, Bartłomiej; Shrier, Alvin; Glass, Leon

2013-01-01

Interactions of a spiral wave with a pacemaker is studied in a mathematical model of two dimensional excitable medium. Faster pacemakers emitting target waves can abolish spirals by driving them to the border of the medium. Our study shows that a slower pacemaker can modify spiral wave behavior by changing the motion of the spiral core. We analyze the dynamics of the spiral wave near the spiral core and away from the core as a function of size and period of the pacemaker. The pacemaker can cause the spiral wave to drift towards it, and either speed up or slow down the reentrant activity. Furthermore, the drift induced by the pacemaker can result in irregular or quasiperiodic dynamics even at sites away from the pacemaker. These results highlight the influence of pacemakers on complex spiral wave dynamics. (paper)

Risk of contrast-medium-induced nephropathy in high-risk patients undergoing MDCT - A pooled analysis of two randomized trials

Energy Technology Data Exchange (ETDEWEB)

Thomsen, Henrik S. [University of Copenhagen, Department of Diagnostic Radiology, Copenhagen University Hospital Herlev, and Department of Diagnostic Sciences, Faculty of Health Sciences, Herlev (Denmark); Morcos, Sameh K. [University of Sheffield, Department of Diagnostic Imaging, Northern General Hospital, Sheffield (United Kingdom)

2009-04-15

The incidence of contrast-medium-induced nephropathy (CIN) following intravenous (IV) CM administration of contrast media to renally impaired patients undergoing multidetector computed tomography (MDCT) is not well characterized. Our objective was to investigate the incidence of CIN in patients with glomerular filtration rate (GFR) <60 ml/min undergoing contrast-enhanced MDCT examinations and to compare the rates of CIN following the IV administration of low-osmolar contrast media (LOCM, iopamidol and iomeprol) and an iso-osmolar contrast medium (IOCM, iodixanol). A total of 301 adult patients with moderate-to-severe renal failure received a similar IV contrast dose (40 gI). Serum creatinine (SCr) was measured at screening, baseline and 48-72 {+-} 6 h after the MDCT examination. Primary CIN outcome was an increase in SCr {>=}0.5 mg/dl ({>=}44.2 {mu}mol/l) from baseline. The CIN rates were 2.3% in the total population, 0.6% when GFR >40 ml/min, 4.6% when GFR <40 ml/min and 7.8% in patients with GFR <30 ml/min. The incidence of CIN was significantly higher after iodixanol than after LOCM (seven patients, 4.7% following IOCM, no CIN cases following the LOCM; p = 0.007). Significant differences in favor of the LOCM were also observed in patients with GFR <40 ml/min and GFR <30 ml/min. Following the IV administration of nonionic contrast agents in patients with moderate-to-severe renal insufficiency, the risk of significant CIN seems to be low. The IOCM iodixanol caused a higher rate of CIN than the LOCM iopamidol and iomeprol, especially in high-risk patients. Differences in osmolality between these LOCM and iodixanol do not play a role in the genesis of CIN. (orig.)

Prevotella intermedia induces severe bacteremic pneumococcal pneumonia in mice with upregulated platelet-activating factor receptor expression.

Science.gov (United States)

Nagaoka, Kentaro; Yanagihara, Katsunori; Morinaga, Yoshitomo; Nakamura, Shigeki; Harada, Tatsuhiko; Hasegawa, Hiroo; Izumikawa, Koichi; Ishimatsu, Yuji; Kakeya, Hiroshi; Nishimura, Masaharu; Kohno, Shigeru

2014-02-01

Streptococcus pneumoniae is the leading cause of respiratory infection worldwide. Although oral hygiene has been considered a risk factor for developing pneumonia, the relationship between oral bacteria and pneumococcal infection is unknown. In this study, we examined the synergic effects of Prevotella intermedia, a major periodontopathic bacterium, on pneumococcal pneumonia. The synergic effects of the supernatant of P. intermedia (PiSup) on pneumococcal pneumonia were investigated in mice, and the stimulation of pneumococcal adhesion to human alveolar (A549) cells by PiSup was assessed. The effects of PiSup on platelet-activating factor receptor (PAFR) transcript levels in vitro and in vivo were analyzed by quantitative real-time PCR, and the differences between the effects of pneumococcal infection induced by various periodontopathic bacterial species were verified in mice. Mice inoculated with S. pneumoniae plus PiSup exhibited a significantly lower survival rate, higher bacterial loads in the lungs, spleen, and blood, and higher inflammatory cytokine levels in the bronchoalveolar lavage fluid (macrophage inflammatory protein 2 and tumor necrosis factor alpha) than those infected without PiSup. In A549 cells, PiSup increased pneumococcal adhesion and PAFR transcript levels. PiSup also increased lung PAFR transcript levels in mice. Similar effects were not observed in the supernatants of Porphyromonas gingivalis or Fusobacterium nucleatum. Thus, P. intermedia has the potential to induce severe bacteremic pneumococcal pneumonia with enhanced pneumococcal adhesion to lower airway cells.

[Detection of fps tumor antigen with mono-specific anti-fps serum in tumors induced by acute transforming ALV].

Science.gov (United States)

Wang, Yixin; Chen, Hao; Zhao, Peng; Li, Jianliang; Cui, Zhizhong

2013-03-04

To prepare anti-fps mono-specific serum, and detect the fps antigen in tumors induced by acute transforming avian leukosis/sarcoma virus containing v-fps oncogene. Two part of v-fps gene was amplified by RT-PCR using the Fu-J viral RNA as the template. Mono-specific serum was prepared by immuning Kunming white mouse with both two recombinant infusion proteins expressed by the prokaryotic expression system. Indirect immunofluorescent assay was used to detect fps antigen in tumor tissue suspension cells and CEF infected by sarcoma supernatant. Immunohistochemical method was used to detect fps antigen in tumor tissue. The mouse mono-specific serum was specific as it had no cross reaction with classical ALV-J strains. The result reveals that the tumor tissue suspension cells, the CEF infected by sarcoma supernatant, and the slice immunohistochemistry of the sarcoma showed positive results. The anti-fps mono-specific serum was prepared, and the detection method was established, which laid the foundation for the study of viral biological characteristics and mechanism of tumourgenesis of acute transforming avian leukosis/sarcoma virus containing v-fps oncogene.

Influence of medium components on the expression of recombinant lipoproteins in Escherichia coli.

Science.gov (United States)

Tseng, Chi-Ling; Leng, Chih-Hsiang

2012-02-01

Bacterial lipoproteins are crucial antigens for protective immunity against bacterial pathogens. Expression of exogenous lipoproteins in Escherichia coli at high levels is thought to be an extremely difficult endeavor because it frequently results in incomplete or absent lipid modification. Previously, we identified a fusion sequence (D1) from a Neisseria meningitidis lipoprotein that induced a non-lipidated protein, E3 (the domain III of the dengue virus envelope protein), to become lipidated. However, without optimizing the growth conditions, some of the D1-fusion proteins were not lipidated. Here, we report the influence of medium components on the expression of recombinant lipoproteins in E. coli. For high-level expression of mature lipoproteins in the C43 (DE3) strain, M9 medium was better than M63 and the rich medium. Furthermore, we analyzed the influence of other media factors (including nitrogen and carbon sources, phosphate, ferrous ions, calcium, magnesium, and pH) on the levels of lipoprotein expression. The results showed that excess nitrogen sources and phosphate in M9 medium could increase the amount of immature lipoproteins, and glucose was a better carbon source than glycerol for expressing mature lipoproteins. We also found that lipoproteins tended to be completely processed in the alkaline environment, even in the nutrient-rich medium. Additional constructs expressing different immunogens or lipid signal peptides as targets were also utilized, demonstrating that these targets could be expressed as completely mature lipoproteins in the M9 medium but not in the rich medium. Our results provide the useful information for expressing mature exogenous lipoproteins in E. coli.

Risk of contrast-medium-induced nephropathy in high-risk patients undergoing MDCT - A pooled analysis of two randomized trials

International Nuclear Information System (INIS)

Thomsen, Henrik S.; Morcos, Sameh K.

2009-01-01

The incidence of contrast-medium-induced nephropathy (CIN) following intravenous (IV) CM administration of contrast media to renally impaired patients undergoing multidetector computed tomography (MDCT) is not well characterized. Our objective was to investigate the incidence of CIN in patients with glomerular filtration rate (GFR) 40 ml/min, 4.6% when GFR <40 ml/min and 7.8% in patients with GFR <30 ml/min. The incidence of CIN was significantly higher after iodixanol than after LOCM (seven patients, 4.7% following IOCM, no CIN cases following the LOCM; p = 0.007). Significant differences in favor of the LOCM were also observed in patients with GFR <40 ml/min and GFR <30 ml/min. Following the IV administration of nonionic contrast agents in patients with moderate-to-severe renal insufficiency, the risk of significant CIN seems to be low. The IOCM iodixanol caused a higher rate of CIN than the LOCM iopamidol and iomeprol, especially in high-risk patients. Differences in osmolality between these LOCM and iodixanol do not play a role in the genesis of CIN. (orig.)

Contrast Medium Induced Nephropathy after Endovascular Stent Graft Placement: An Examination of Its Prevalence and Risk Factors

International Nuclear Information System (INIS)

Kawatani, Yohei; Nakamura, Yoshitsugu; Mochida, Yoshihiko; Yamauchi, Naoya; Hayashi, Yujiro; Taneichi, Tetsuyoshi; Ito, Yujiro; Kurobe, Hirotsugu; Suda, Yuji; Hori, Takaki

2016-01-01

Endovascular stent graft placement has become a major treatment for thoracic and abdominal aneurysms. While endovascular therapy is less invasive than open surgery, it involves the use of a contrast medium. Contrast media can cause renal impairment, a condition termed as contrast-induced nephropathy (CIN). This study sought to evaluate the incidence and risk factors of CIN following endovascular stent graft placement for aortic aneurysm repair. The study included 167 consecutive patients who underwent endovascular stent graft placement in our hospital from October 2013 to June 2014. CIN was diagnosed using the European Society of Urogenital Radiology criteria. Patients with and without CIN were compared. Chi-squared tests, t-tests, and multivariate logistic regression analyses were performed. Thirteen patients (7.8%) developed CIN. Left ventricular dysfunction and intraoperative blood transfusion were significantly more frequent in the CIN group (P = 0.017 and P = 0.032, resp.). Multivariate analysis showed that left ventricular dysfunction had the strongest influence on CIN development (odds ratio 9.34, P = 0.018, and 95% CI = 1.46–59.7). Patients with CIN also experienced longer ICU and hospital stays. Measures to improve renal perfusion flow should be considered for patients with left ventricular dysfunction who are undergoing endovascular stent graft placement

Pseudomonas aeruginosa toxin ExoU induces a PAF-dependent impairment of alveolar fibrin turnover secondary to enhanced activation of coagulation and increased expression of plasminogen activator inhibitor-1 in the course of mice pneumosepsis

Directory of Open Access Journals (Sweden)

Suassuna José HR

2011-08-01

Full Text Available Abstract Background ExoU, a Pseudomonas aeruginosa cytotoxin with phospholipase A2 activity, was shown to induce vascular hyperpermeability and thrombus formation in a murine model of pneumosepsis. In this study, we investigated the toxin ability to induce alterations in pulmonary fibrinolysis and the contribution of the platelet activating factor (PAF in the ExoU-induced overexpression of plasminogen activator inhibitor-1 (PAI-1. Methods Mice were intratracheally instilled with the ExoU producing PA103 P. aeruginosa or its mutant with deletion of the exoU gene. After 24 h, animal bronchoalveolar lavage fluids (BALF were analyzed and lung sections were submitted to fibrin and PAI-1 immunohistochemical localization. Supernatants from A549 airway epithelial cells and THP-1 macrophage cultures infected with both bacterial strains were also analyzed at 24 h post-infection. Results In PA103-infected mice, but not in control animals or in mice infected with the bacterial mutant, extensive fibrin deposition was detected in lung parenchyma and microvasculature whereas mice BALF exhibited elevated tissue factor-dependent procoagulant activity and PAI-1 concentration. ExoU-triggered PAI-1 overexpression was confirmed by immunohistochemistry. In in vitro assays, PA103-infected A549 cells exhibited overexpression of PAI-1 mRNA. Increased concentration of PAI-1 protein was detected in both A549 and THP-1 culture supernatants. Mice treatment with a PAF antagonist prior to PA103 infection reduced significantly PAI-1 concentrations in mice BALF. Similarly, A549 cell treatment with an antibody against PAF receptor significantly reduced PAI-1 mRNA expression and PAI-1 concentrations in cell supernatants, respectively. Conclusion ExoU was shown to induce disturbed fibrin turnover, secondary to enhanced procoagulant and antifibrinolytic activity during P. aeruginosa pneumosepsis, by a PAF-dependent mechanism. Besides its possible pathophysiological relevance, in

Cross-Kerr nonlinearities in an optically dressed periodic medium

Energy Technology Data Exchange (ETDEWEB)

Slowik, K; Raczynski, A; Zaremba, J [Instytut Fizyki, Uniwersytet Mikolaja Kopernika, ulica Grudziadzka 5, 87-100 Torun (Poland); Zielinska-Kaniasty, S [Instytut Matematyki i Fizyki, Uniwersytet Technologiczno-Przyrodniczy, Aleja Prof. S Kaliskiego 7, 85-789 Bydgoszcz (Poland); Artoni, M [Department of Physics and Chemistry of Materials, CNR-INFM Sensor Lab, Brescia University and European Laboratory for Nonlinear Spectroscopy, Firenze (Italy); La Rocca, G C, E-mail: karolina@fizyka.umk.pl [Scuola Normale Superiore and CNISM, Pisa (Italy)

2011-02-15

Cross-Kerr nonlinearities are analyzed for two light beams propagating in an atomic medium in the tripod configuration, dressed by a strong standing-wave laser field that induces periodic optical properties. The reflection and transmission spectra as well as the phases of both the reflected and transmitted components of the two beams are analyzed theoretically with nonlinearities up to third order being taken into account. Ranges of parameters are sought in which the cross-Kerr effect can be used as the basis of the phase gate.

Impact of external medium conductivity on cell membrane electropermeabilization by microsecond and nanosecond electric pulses

Science.gov (United States)

Silve, Aude; Leray, Isabelle; Poignard, Clair; Mir, Lluis M.

2016-01-01

The impact of external medium conductivity on the efficiency of the reversible permeabilisation caused by pulsed electric fields was investigated. Pulses of 12 ns, 102 ns or 100 μs were investigated. Whenever permeabilisation could be detected after the delivery of one single pulse, media of lower conductivity induced more efficient reversible permeabilisation and thus independently of the medium composition. Effect of medium conductivity can however be hidden by some saturation effects, for example when pulses are cumulated (use of trains of 8 pulses) or when the detection method is not sensitive enough. This explains the contradicting results that can be found in the literature. The new data are complementary to those of one of our previous study in which an opposite effect of the conductivity was highlighted. It stresses that the conductivity of the medium influences the reversible permeabilization by several ways. Moreover, these results clearly indicate that electropermeabilisation does not linearly depend on the energy delivered to the cells. PMID:26829153

Protective effect of lycopene for oxidative damage in human lens epithelial cells induced by UV

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Jing-Wen Sun

2016-05-01

Full Text Available AIM:To investigate the protective effect and possible mechanisms of lycopene for oxidative damage induced by ultraviolet in cultured human lens epithelial cells(HLEC. METHODS:HLEC was subcultured and divided into negative control group, oxidative injury group, lycopene low dose group and lycopene high dose group. Cell viability was assayed by MTT colorimetric. Cell morphological changes were detected by electron microscope. Reactive oxygen species(ROSlevels were detected with DCFH-DA fluorescent probe. Content of superoxide dismutase(SOD, glutathione peroxidase(GSHand malondialdehyde(MDAin supernatants were detected by spectrophotometer. RESULTS:Lycopene could obviously inhibited UV-induced decline in cell activity, reduce UV-induced ROS generation within HLEC, cause SOD, GSH-Px levels increased and MDA levels decreased.CONCLUSION:Lycopene plays its strong antioxidant role in increasing the intracellular SOD and GSH-Px content levels and decreasing MDA levels, which provide reliable experimental basis for prevent and treatment of cataracts.

Pencapaian Fase Embriosomatik Manggis (Garcinia mangostana L. dengan Penambahan Thidiazuron dalam Medium Setengah MS Cair

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Innaka Ageng Rineksane

2016-02-01

Full Text Available One of the problems related to the establishment of mangosteen plantation is to obtain seedlings throughout the year, which can be solved by micropropagation.   The propagation of Mangosteen was done through somatic embryogenesis. The objective of this study was to determine the effect of Thidiazuron concentration in ½ MS medium to achieve somatic embryo stages of mangosteen seed. The study consisted of two experiments. Experiment 1: The embryogenic callus c.a.1 g derived from MS solid medium containing of Thidiazuron (0,1; 0,5; dan 1 mg/l and 2,4-Dichlorophenoxy acetic acid (4,6,8 dan 10 mg/l was subcultured into ½ MS0 liquid medium. 2. The embryogenic callus from the previous medium were subcultured into ½ MS liquid medium containing Thidiazuron (0, 1, 2, 4 dan 8 mg/L and Casein hydrolysate 500 mg/L. The results showed that cell suspension were developed after mangosteen embryogenic callus subcultured into ½ MS0 liquid medium.  Somatic embryo stages such as globular, heart and torpedo were formed after the calli derived from the previous ½ MS solid medium containing 10 mg/L 2,4-D and 1 mg/L Thidiazuron were subcultured into ½ MS0 liquid medium in 8 weeks.  Cell suspension in ½ MS liquid medium with or without the addition of  Thidiazuron and Casein hydrolysate has induced the formation of embryo somatic stages such as globular, heart and torpedo after 6 months of incubation.

Near infrared spectroscopy combined with multivariate analysis for monitoring the ethanol precipitation process of fraction I + II + III supernatant in human albumin separation

Science.gov (United States)

Li, Can; Wang, Fei; Zang, Lixuan; Zang, Hengchang; Alcalà, Manel; Nie, Lei; Wang, Mingyu; Li, Lian

2017-03-01

Nowadays, as a powerful process analytical tool, near infrared spectroscopy (NIRS) has been widely applied in process monitoring. In present work, NIRS combined with multivariate analysis was used to monitor the ethanol precipitation process of fraction I + II + III (FI + II + III) supernatant in human albumin (HA) separation to achieve qualitative and quantitative monitoring at the same time and assure the product's quality. First, a qualitative model was established by using principal component analysis (PCA) with 6 of 8 normal batches samples, and evaluated by the remaining 2 normal batches and 3 abnormal batches. The results showed that the first principal component (PC1) score chart could be successfully used for fault detection and diagnosis. Then, two quantitative models were built with 6 of 8 normal batches to determine the content of the total protein (TP) and HA separately by using partial least squares regression (PLS-R) strategy, and the models were validated by 2 remaining normal batches. The determination coefficient of validation (Rp2), root mean square error of cross validation (RMSECV), root mean square error of prediction (RMSEP) and ratio of performance deviation (RPD) were 0.975, 0.501 g/L, 0.465 g/L and 5.57 for TP, and 0.969, 0.530 g/L, 0.341 g/L and 5.47 for HA, respectively. The results showed that the established models could give a rapid and accurate measurement of the content of TP and HA. The results of this study indicated that NIRS is an effective tool and could be successfully used for qualitative and quantitative monitoring the ethanol precipitation process of FI + II + III supernatant simultaneously. This research has significant reference value for assuring the quality and improving the recovery ratio of HA in industrialization scale by using NIRS.

Near infrared spectroscopy combined with multivariate analysis for monitoring the ethanol precipitation process of fraction I+II+III supernatant in human albumin separation.

Science.gov (United States)

Li, Can; Wang, Fei; Zang, Lixuan; Zang, Hengchang; Alcalà, Manel; Nie, Lei; Wang, Mingyu; Li, Lian

2017-03-15

Nowadays, as a powerful process analytical tool, near infrared spectroscopy (NIRS) has been widely applied in process monitoring. In present work, NIRS combined with multivariate analysis was used to monitor the ethanol precipitation process of fraction I+II+III (FI+II+III) supernatant in human albumin (HA) separation to achieve qualitative and quantitative monitoring at the same time and assure the product's quality. First, a qualitative model was established by using principal component analysis (PCA) with 6 of 8 normal batches samples, and evaluated by the remaining 2 normal batches and 3 abnormal batches. The results showed that the first principal component (PC1) score chart could be successfully used for fault detection and diagnosis. Then, two quantitative models were built with 6 of 8 normal batches to determine the content of the total protein (TP) and HA separately by using partial least squares regression (PLS-R) strategy, and the models were validated by 2 remaining normal batches. The determination coefficient of validation (R p 2 ), root mean square error of cross validation (RMSECV), root mean square error of prediction (RMSEP) and ratio of performance deviation (RPD) were 0.975, 0.501g/L, 0.465g/L and 5.57 for TP, and 0.969, 0.530g/L, 0.341g/L and 5.47 for HA, respectively. The results showed that the established models could give a rapid and accurate measurement of the content of TP and HA. The results of this study indicated that NIRS is an effective tool and could be successfully used for qualitative and quantitative monitoring the ethanol precipitation process of FI+II+III supernatant simultaneously. This research has significant reference value for assuring the quality and improving the recovery ratio of HA in industrialization scale by using NIRS. Copyright © 2016 Elsevier B.V. All rights reserved.

The difference in noise property between the Autler—Townes splitting medium and the electromagnetically induced transparent medium

International Nuclear Information System (INIS)

Li Zhong-Hua; Li Yuan; Dou Ya-Fang; Zhang Jun-Xiang

2012-01-01

The quantum noise of squeezed probe light passing through an atomic system with different electromagnetically induced transparency and Autler—Townes splitting effects is investigated theoretically. It is found that the optimal squeezing preservation of the outgoing probe beam occurs in the strong-coupling-field regime rather than in the weak-coupling-field regime. In the weak-coupling-field regime, which was recently recognized as the electromagnetically induced transparency regime (Abi-Salloum T Y 2010 Phys. Rev. A 81 053836), the output amplitude noise is affected mainly by the atomic noise originating from the random decay process of atoms. While in the strong-coupling-field regime, defined as the Autler—Townes splitting regime, the output amplitude noise is affected mainly by the phase-to-amplitude conversion noise. This is useful in improving the quality of the experiment for efficient quantum memory, and hence has an application in quantum information processing. (electromagnetism, optics, acoustics, heat transfer, classical mechanics, and fluid dynamics)

Medium-chain fatty acids as ligands for orphan G protein-coupled receptor GPR84.

Science.gov (United States)

Wang, Jinghong; Wu, Xiaosu; Simonavicius, Nicole; Tian, Hui; Ling, Lei

2006-11-10

Free fatty acids (FFAs) play important physiological roles in many tissues as an energy source and as signaling molecules in various cellular processes. Elevated levels of circulating FFAs are associated with obesity, dyslipidemia, and diabetes. Here we show that GPR84, a previously orphan G protein-coupled receptor, functions as a receptor for medium-chain FFAs with carbon chain lengths of 9-14. Medium-chain FFAs elicit calcium mobilization, inhibit 3',5'-cyclic AMP production, and stimulate [35S]guanosine 5'-O-(3-thiotriphosphate) binding in a GPR84-dependent manner. The activation of GPR84 by medium-chain FFAs couples primarily to a pertussis toxin-sensitive G(i/o) pathway. In addition, we show that GPR84 is selectively expressed in leukocytes and markedly induced in monocytes/macrophages upon activation by lipopolysaccharide. Furthermore, we demonstrate that medium-chain FFAs amplify lipopolysaccharide-stimulated production of the proinflammatory cytokine interleukin-12 p40 through GPR84. Our results indicate a role for GPR84 in directly linking fatty acid metabolism to immunological regulation.

Live and Heat-Killed Lactobacillus rhamnosus ATCC 7469 May Induce Modulatory Cytokines Profiles on Macrophages RAW 264.7

OpenAIRE

Jorj?o, Adeline Lacerda; de Oliveira, Felipe Eduardo; Le?o, Mariella Vieira Pereira; Carvalho, Cl?udio Antonio Talge; Jorge, Antonio Olavo Cardoso; de Oliveira, Luciane Dias

2015-01-01

This study aimed to evaluate the capacity of Lactobacillus rhamnosus and/or its products to induce the synthesis of cytokines (TNF-?, IL-1?, IL-4, IL-6, IL-10, and IL-12) by mouse macrophages (RAW 264.7). Three microorganism preparations were used: live L. rhamnosus (LLR) suspension, heat-killed L. rhamnosus (HKLR) suspension, and the supernatant of a heat-killed L. rhamnosus (SHKLR) suspension, which were cultured with macrophages (37?C, 5% CO2) for 2?h and 30?min. After that, cells were cul...

Plasma generated in culture medium induces damages of HeLa cells due to flow phenomena

Science.gov (United States)

Sato, Yusuke; Sato, Takehiko; Yoshino, Daisuke

2018-03-01

Plasma in a liquid has been anticipated as an effective tool for medical applications, however, few reports have described cellular responses to plasma generated in a liquid similar to biological fluids. Herein we report the effects of plasma generated in a culture medium on HeLa cells. The plasma in the culture medium produced not only heat, shock waves, and reactive chemical species but also a jet flow with sub millimeter-sized bubbles. Cells exposed to the plasma exhibited detachment, morphological changes, and changes in the actin cytoskeletal structure. The experimental results suggest that wall shear stress over 160 Pa was generated on the surface of the cells by the plasma. It is one of the main factors that cause those cellular responses. We believe that our findings would provide valuable insight into advancements in medical applications of plasma in a liquid.

Prospective study of pathogens in asymptomatic travellers and those with diarrhoea: aetiological agents revisited.

Science.gov (United States)

Lääveri, T; Antikainen, J; Pakkanen, S H; Kirveskari, J; Kantele, A

2016-06-01

Travellers' diarrhoea (TD) remains the most frequent health problem encountered by visitors to the (sub)tropics. Traditional stool culture identifies the pathogen in only 15% of cases. Exploiting PCR-based methods, we investigated TD pathogens with a focus on asymptomatic travellers and severity of symptoms. Pre- and post-travel stools of 382 travellers with no history of antibiotic use during travel were analysed with a multiplex quantitative PCR for Salmonella, Yersinia, Campylobacter, Shigella, Vibrio cholerae and five diarrhoeagenic Escherichia coli: enteroaggregative (EAEC), enteropathogenic (EPEC), enterotoxigenic (ETEC), enterohaemorrhagic (EHEC) and enteroinvasive (EIEC). The participants were categorized by presence/absence of TD during travel and on return, and by severity of symptoms. A pathogen was indentified in 61% of the asymptomatic travellers, 83% of those with resolved TD, and 83% of those with ongoing TD; 25%, 43% and 53% had multiple pathogens, respectively. EPEC, EAEC, ETEC and Campylobacter associated especially with ongoing TD symptoms. EAEC and EPEC proved more common than ETEC. To conclude, modern methodology challenges our perception of stool pathogens: all pathogens were common both in asymptomatic and symptomatic travellers. TD has a multibacterial nature, but diarrhoeal symptoms mostly associate with EAEC, EPEC, ETEC and Campylobacter. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.

Effect of Lactobacillus rhamnosus GG supernatant on serotonin transporter expression in rats with post-infectious irritable bowel syndrome

Science.gov (United States)

Cao, Ya-Nan; Feng, Li-Juan; Liu, Yuan-Yuan; Jiang, Kui; Zhang, Mao-Jun; Gu, Yi-Xin; Wang, Bang-Mao; Gao, Jia; Wang, Ze-Lan; Wang, Yu-Ming

2018-01-01

AIM To evaluate the effect of Lactobacillus rhamnosus GG supernatant (LGG-s) on the expression of serotonin transporter (SERT) in rats with post-infectious irritable bowel syndrome (PI-IBS). METHODS Campylobacter jejuni 81-176 (1010 CFU/mL) was used to induce intestinal infection to develop a PI-IBS model. After evaluation of the post-infectious phase by biochemical tests, DNA agarose gel electrophoresis, abdominal withdrawal reflex (AWR) test, and the intestinal motility test, four PI-IBS groups received different concentrations of LGG-s for 4 wk. The treatments were maintained for 1.0, 2.0, 3.0 or 4.0 wk during the experiment, and the colons and brains were removed for later use each week. SERT mRNA and protein levels were detected by real-time PCR and Western blot, respectively. RESULTS The levels of SERT mRNA and protein in intestinal tissue were higher in rats treated with LGG-s than in control rats and PI-IBS rats gavaged with PBS during the whole study. Undiluted LGG-s up-regulated SERT mRNA level by 2.67 times compared with the control group by week 2, and SERT mRNA expression kept increasing later. Double-diluted LGG-s was similar to undiluted-LGG-s, resulting in high levels of SERT mRNA. Triple-diluted LGG-s up-regulated SERT mRNA expression level by 6.9-times compared with the control group, but SERT mRNA expression decreased rapidly at the end of the second week. At the first week, SERT protein levels were basically comparable in rats treated with undiluted LGG-s, double-diluted LGG-s, and triple-diluted LGG-s, which were higher than those in the control group and PBS-treated PI-IBS group. SERT protein levels in the intestine were also comparable in rats treated with undiluted LGG-s, double-diluted LGG-s, and triple-diluted LGG-s by the second and third weeks. SERT mRNA and protein levels in the brain had no statistical difference in the groups during the experiment. CONCLUSION LGG-s can up-regulate SERT mRNA and protein levels in intestinal tissue but

Determinants of Plant Growth-promoting Ochrobactrum lupini KUDC1013 Involved in Induction of Systemic Resistance against Pectobacterium carotovorum subsp. carotovorum in Tobacco Leaves

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Marilyn Sumayo

2013-06-01

Full Text Available The plant growth-promoting rhizobacterium Ochrobactrum lupini KUDC1013 elicited induced systemic resistance (ISR in tobacco against soft rot disease caused by Pectobacterium carotovorum subsp. carotovorum. We investigated of its factors involved in ISR elicitation. To characterize the ISR determinants, KUDC1013 cell suspension, heat-treated cells, supernatant from a culture medium, crude bacterial lipopolysaccharide (LPS and flagella were tested for their ISR activities. Both LPS and flagella from KUDC1013 were effective in ISR elicitation. Crude cell free supernatant elicited ISR and factors with the highest ISR activity were retained in the n-butanol fraction. Analysis of the ISR-active fraction revealed the metabolites, phenylacetic acid (PAA, 1-hexadecene and linoleic acid (LA, as elicitors of ISR. Treatment of tobacco with these compounds significantly decreased the soft rot disease symptoms. This is the first report on the ISR determinants by plant growth-promoting rhizobacteria (PGPR KUDC1013 and identifying PAA, 1-hexadecene and LA as ISR-related compounds. This study shows that KUDC1013 has a great potential as biological control agent because of its multiple factors involved in induction of systemic resistance against phytopathogens.

Nonlinear radiative peristaltic flow of hydromagnetic fluid through porous medium

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Q. Hussain

2018-06-01

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

Narrowing of electromagnetically induced transparency resonance in a Doppler-broadened medium

International Nuclear Information System (INIS)

Javan, Ali; Kocharovskaya, Olga; Lee Hwang; Scully, Marlan O.

2002-01-01

We derive an analytic expression for the linewidth of electromagnetically induced transparency (EIT) resonance in a Doppler-broadened system. It is shown here that for relatively low intensity of the driving field the EIT linewidth is proportional to the square root of intensity and is independent of the Doppler width, similar to the laser-induced line narrowing effect described by Feld and Javan. In the limit of high intensity we recover the usual power-broadening case where the EIT linewidth is proportional to the intensity and inversely proportional to the Doppler width

Spectral Induced Polarization Signatures of Ethanol in Sand-Clay Medium

Science.gov (United States)

The spectral Induced Polarization (SIP) method has previously been investigated as a tool for detecting physicochemical changes occurring as result of clay-organic interactions in porous media. We performed SIP measurements with a dynamic signal analyzer (NI-4551) on laboratory ...

The effect of aprotinin on hypoxia-reoxygenation-induced changes in neutrophil and endothelial function.

LENUS (Irish Health Repository)

Harmon, D

2012-02-03

BACKGROUND AND OBJECTIVE: An acute inflammatory response associated with cerebral ischaemia-reperfusion contributes to the development of brain injury. Aprotinin has potential, though unexplained, neuroprotective effects in patients undergoing cardiac surgery. METHODS: Human neutrophil CD11 b\\/CD18, endothelial cell intercellular adhesion molecule-1 (ICAM-1) expression and endothelial interleukin (IL)-1beta supernatant concentrations in response to in vitro hypoxia-reoxygenation was studied in the presence or absence of aprotinin (1600 KIU mL(-1)). Adhesion molecule expression was quantified using flow cytometry and IL-1beta concentrations by enzyme-linked immunosorbent assay. Data were analysed using ANOVA and post hoc Student-Newman-Keuls test as appropriate. RESULTS: Exposure to 60-min hypoxia increased neutrophil CD11b expression compared to normoxia (170+\\/-46% vs. 91+\\/-27%, P = 0.001) (percent intensity of fluorescence compared to time 0) (n = 8). Hypoxia (60 min) produced greater upregulation of CD11b expression in controls compared to aprotinin-treated neutrophils [(170+\\/-46% vs. 129+\\/-40%) (P = 0.04)] (n = 8). Hypoxia-reoxygenation increased endothelial cell ICAM-1 expression (155+\\/-3.7 vs. 43+\\/-21 mean channel fluorescence, P = 0.0003) and IL-1beta supernatant concentrations compared to normoxia (3.4+\\/-0.4 vs. 2.6+\\/-0.2, P = 0.02) (n = 3). Hypoxia-reoxygenation produced greater upregulation of ICAM- 1 expression [(155+\\/-3.3 vs. 116+\\/-0.7) (P = 0.001)] and IL-1beta supernatant concentrations [(3.4+\\/-0.3 vs. 2.6+\\/-0.1) (P = 0.01)] in controls compared to aprotinin-treated endothelial cell preparation (n = 3). CONCLUSIONS: Hypoxia-reoxygenation-induced upregulation of neutrophil CD11b, endothelial cell ICAM-1 expression and IL-1beta concentrations is decreased by aprotinin at clinically relevant concentrations.

Effects and mechanisms of total glucosides of paeony on joint damage in rat collagen-induced arthritis.

Science.gov (United States)

Zhu, L; Wei, W; Zheng, Y-Q; Jia, X-Y

2005-05-01

To investigate the therapeutic effects and mechanisms of total glucosides of paeony (TGP), an effective compound of Chinese traditional herbal medicine (CTM), on collagen -induced arthritis (CIA) in rats. CIA was induced in male Sprague-Dawley rats immunized with chicken type II collagen in Freund's complete adjuvant. TGP (25, 50, 100 mg/kg/d) was orally administered to rats from day 14 to 28 after immunization. Arthritis was evaluated by hind paw swelling, polyarthritis index, and histological examination. Activities of interleukin-1 (IL-1) and tumor necrosis factor alpha (TNFalpha) were determined and the ultrastructure of synoviocytes was observed. The proliferation and the production of vascular epidermal growth factor (VEGF), basic fibroblast growth factor (bFGF), matrix metalloproteinase 1 (MMP-1) and MMP-3 in fibroblast-like synoviocytes (FLS) were detected. The administration of TGP (25, 50, 100 mg/kg, ig x 14 days) suppressed secondary inflammatory reactions and histological changes in CIA model. The ultrastructure of synoviocytes from CIA rats was changed, and the level of IL-1 and TNF alpha produced by macrophage-like synoviocytes (MLS) from CIA rats was elevated. TGP (50, 100 mg/kg, ig x 14 days) inhibited above changes significantly. The MLS supernatants of CIA rats induced more cell proliferation and more production of VEGF, bFGF, MMP-1 and MMP-3 in FLS of CIA than those supernatants from CIA rats treated with TGP (50, 100 mg/kg, ig x 14 days). These results indicate that TGP exerts a suppressive effect on joint destruction in rat CIA. The therapeutic effect of TGP could be associated with its ability to ameliorate the secretion and metabolism of synoviocytes and to inhibit the abnormal proliferation and VEGF, bFGF, MMP-1 and MMP-3 production by FLS.

NF-κB dependent and independent mechanisms of quartz-induced proinflammatory activation of lung epithelial cells

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Schins Roel PF

2010-05-01

Full Text Available Abstract In the initiation and progression of pulmonary inflammation, macrophages have classically been considered as a crucial cell type. However, evidence for the role of epithelial type II cells in pulmonary inflammation has been accumulating. In the current study, a combined in vivo and in vitro approach has been employed to investigate the mechanisms of quartz-induced proinflammatory activation of lung epithelial cells. In vivo, enhanced expression of the inflammation- and oxidative stress-related genes HO-1 and iNOS was found on the mRNA level in rat lungs after instillation with DQ12 respirable quartz. Activation of the classical NF-κB pathway in macrophages and type II pneumocytes was indicated by enhanced immunostaining of phospho-IκBα in these specific lung cell types. In vitro, the direct, particle-mediated effect on proinflammatory signalling in a rat lung epithelial (RLE cell line was compared to the indirect, macrophage product-mediated effect. Treatment with quartz particles induced HO-1 and COX-2 mRNA expression in RLE cells in an NF-κB independent manner. Supernatant from quartz-treated macrophages rapidly activated the NF-κB signalling pathway in RLE cells and markedly induced iNOS mRNA expression up to 2000-fold compared to non-treated control cells. Neutralisation of TNFα and IL-1β in macrophage supernatant did not reduce its ability to elicit NF-κB activation of RLE cells. In addition the effect was not modified by depletion or supplementation of intracellular glutathione. The results from the current work suggest that although both oxidative stress and NF-κB are likely involved in the inflammatory effects of toxic respirable particles, these phenomena can operate independently on the cellular level. This might have consequences for in vitro particle hazard testing, since by focusing on NF-κB signalling one might neglect alternative inflammatory pathways.

Low dose radiation induced protein and its effect on expression of CD25 molecule in lymphocytes

International Nuclear Information System (INIS)

Lu Duicai; Su Liaoyuan

2001-01-01

Objective: To find the substantial basis for effects of low dose radiation, on development, extraction, and the biogical activity of the low-dose radiation-induced proteins, and the effects of LDR induced proteins on CD25 molecule expression of human lymphocytes. Methods: 1. Healthy Kumning male mice exposed to radiation of 226 Ra γ-rays at 5, 10 and 15 cGy respectively. The mice were killed 2 hours after exposure, the spleen cells were broken with ultrasonic energy and then ultra-centrifugalized at low temperature (4 degree C). The LDR-induced proteins were obtained in the supernatant solution. Then the changes of CD25 molecule was measured by flow cytometry (FCM) with immunofluorescence technique, which was used to reflect the effect of LDR induced proteins on CD25 molecule expression of human lymphocytes. Results: LDR induced proteins were obtained from spleen cells in mice exposed to 5-15 cGy whole body radiation. Conclusion: The expression of CD25 molecule of lymphocytes was increased significantly after use of LDR induced proteins. LDR induced proteins can enhance expression of CD25 molecule of lymphocytes slightly

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

International Nuclear Information System (INIS)

Dalla Costa, C.

2007-07-01

We try to identify and model physical and chemical mechanisms governing the water flow and the solute transport in fractured consolidated porous medium. An original experimental device was built. The 'cube' consists of an idealized fractured medium reproduced by piling up consolidated porous cubes of 5 cm edge. Meanwhile, columns of the homogeneous consolidated porous medium are studied. The same anionic tracing technique is used in both cases. Using a system analysis approach, we inject concentration pulses in the device to obtain breakthrough curves. After identifying the mass balance and the residence time, we fit the CD and the MIM models to the experimental data. The MIM model is able to reproduce experimental curves of the homogeneous consolidated porous medium better than the CD model. The mobile water fraction is in accordance with the porous medium geometry. The study of the flow rate influence highlights an interference dispersion regime. It was not possible to highlight the observation length influence in this case. On the contrary, we highlight the effect of the observation scale on the fractured and porous medium, comparing the results obtained on a small 'cube' and a big 'cube'. The CD model is not satisfactory in this case. Even if the MIM model can fit the experimental breakthrough curves, it was not possible to obtain unique parameters for the set of experiments. (author)

Randomized clinical trial of mast cell inhibition in patients with a medium-sized abdominal aortic aneurysm

DEFF Research Database (Denmark)

Sillesen, H; Eldrup, N; Hultgren, R

2015-01-01

the growth of medium-sized AAAs. In preclinical and clinical trials, pemirolast has been shown to inhibit antigen-induced allergic reactions. METHODS: Inclusion criteria for the trial were patients with an AAA of 39-49 mm in diameter on ultrasound imaging. Among exclusion criteria were previous aortic....... There was no statistically significant difference in growth between patients receiving placebo and those in the three dose groups of pemirolast. Similarly, there were no differences in adverse events. CONCLUSION: Treatment with pemirolast did not retard the growth of medium-sized AAAs. REGISTRATION NUMBER: NCT01354184...

Process and apparatus for reacting laser radiation with a reactive medium

International Nuclear Information System (INIS)

Vanderleeden, J.C.

1980-01-01

The invention is based on the concept of irradiating the reaction medium with laser radiation in a reaction zone bounded by two longitudinally spaced reflecting surfaces, the beam of laser radiation being reflected back and forth between the surfaces which are contoured in such a way that the radiation flux density profile at substantially all transverse cross sections of the reaction zone is matched to the transverse distribution profile, that is the transverse distribution of the availability of a reactive species in the medium. A necessary condition for achieving this is that the beam be successively reflected between the surfaces along successive paths of progressively changing cross-sectional area intersected by respective, contiguous, non-overlapping areas of these surfaces. This process may be applied in particular to the selective laser-induced decomposition of HDCO to yield HD and CO

POTENSI PROBIOTIK INDIGENUS Lactobacillus plantarum Dad 13 PADA YOGURT DENGAN SUPLEMENTASI EKSTRAK UBI JALAR UNGU UNTUK PENURUN DIARE DAN RADIKAL BEBAS

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Agustina Intan Niken Tari

2016-04-01

Full Text Available The purpose of this study was studying the effectiveness of selected indigenous probiotic strains (Lactobacillus plantarum Dad 13 in yoghurt with purple sweet potato extract suplementation as reducing diarrhea and free radicals on white rats albino Norway rats (Rattus novergicus Sprague dawley strain. The study was designed using factorial completely randomized design, with treatment of purple sweet potato extract yogurt without probiotics (P0, purple sweet potato extract yogurt with probiotic (P1 to 2 groups of male Sprague dawley rats were treated without Enteropathogenik Escherichia coli (EPEC ATCC 35218 (E0 and with Enteropathogenik Escherichia coli (EPEC ATCC 35218 (E1. Probiotic treatment was conducted using the sonde at day 1st to 21st at a dose of 1 ml / 120 g weight or average 109 CFU/ ml. While the treatment of EPEC ATCC 35218 was conducted using the sonde at dose of 106 CFU/ml on day 7th to 14th. The observed parameters include fecal water content, water content of cecum, malonaldehide levels (MDA blood and liver. The results showed that (1 There was interaction between the treatment of indigenous probiotic yogurt purple sweet potato extract and EPEC ATCC 35218 on water content of faecal, water content of cecum, MDA levels blood and liver (2.Culture of Lactobacillus plantarum Dad 13 was able to provide health effects as reducing of diarrhea and free radicals. Keywords: Reducing of diarrhea, free radicals, purple sweet potato extract yogurt, probiotic bacteria   ABSTRAK Penelitian bertujuan mempelajari efektivitas strain probiotik indigenus terpilih (Lactobacillus plantarum Dad 13 pada yogurt dengan suplementasi ekstrak ubi jalar ungu sebagai penurun diare dan radikal bebas pada tikus putih albino Norway rats (Rattus novergicus galur Sprague dawley. Penelitian dirancang menggunakan rancangan acak lengkap faktorial, dengan perlakuan yogurt ekstrak ubi jalar ungu tanpa probiotik (P0, yogurt ekstrak ubi jalar ungu dengan probiotik (P1

The Interstellar Medium

CERN Document Server

Lequeux, James

2005-01-01

Describing interstellar matter in our galaxy in all of its various forms, this book also considers the physical and chemical processes that are occurring within this matter. The first seven chapters present the various components making up the interstellar matter and detail the ways that we are able to study them. The following seven chapters are devoted to the physical, chemical and dynamical processes that control the behaviour of interstellar matter. These include the instabilities and cloud collapse processes that lead to the formation of stars. The last chapter summarizes the transformations that can occur between the different phases of the interstellar medium. Emphasizing methods over results, "The Interstellar Medium" is written for graduate students, for young astronomers, and also for any researchers who have developed an interest in the interstellar medium.

Computer modelling of radiation-induced bystander effect

International Nuclear Information System (INIS)

Khvostunov, Igor K.; Nikjoo, Hooshang

2002-01-01

Radiation-induced genomic instability and bystander effects are now well established consequences of exposure of living cells to ionising radiation. It has been observed that cells not directly hit by radiation tracks may still exhibit radiation effects. We present a quantitative modelling of the radiation-induced bystander effect based on a diffusion model of spreading the bystander signal. The model assumes the bystander factor to be a protein of low molecular weight, given out by the hit cell, diffusing in the medium and reacting with non-hit cells. The model calculations successfully predict the results of cell survival in an irradiated conditioned medium. The model predicts the shape of dose-effect relationship for cell survival and oncogenic transformation induced by broad-beam and micro-beam irradiation by alpha-particles. (author)

Concept medium programme

DEFF Research Database (Denmark)

Bjerrum, Peter

2005-01-01

The present essay is an attempt to determine the architectural project of the 21st century in relation to a modern conception of space as the medium of architecture, and of sociality as its program......The present essay is an attempt to determine the architectural project of the 21st century in relation to a modern conception of space as the medium of architecture, and of sociality as its program...

Rapid Induction of Cerebral Organoids From Human Induced Pluripotent Stem Cells Using a Chemically Defined Hydrogel and Defined Cell Culture Medium.

Science.gov (United States)

Lindborg, Beth A; Brekke, John H; Vegoe, Amanda L; Ulrich, Connor B; Haider, Kerri T; Subramaniam, Sandhya; Venhuizen, Scott L; Eide, Cindy R; Orchard, Paul J; Chen, Weili; Wang, Qi; Pelaez, Francisco; Scott, Carolyn M; Kokkoli, Efrosini; Keirstead, Susan A; Dutton, James R; Tolar, Jakub; O'Brien, Timothy D

2016-07-01

Tissue organoids are a promising technology that may accelerate development of the societal and NIH mandate for precision medicine. Here we describe a robust and simple method for generating cerebral organoids (cOrgs) from human pluripotent stem cells by using a chemically defined hydrogel material and chemically defined culture medium. By using no additional neural induction components, cOrgs appeared on the hydrogel surface within 10-14 days, and under static culture conditions, they attained sizes up to 3 mm in greatest dimension by day 28. Histologically, the organoids showed neural rosette and neural tube-like structures and evidence of early corticogenesis. Immunostaining and quantitative reverse-transcription polymerase chain reaction demonstrated protein and gene expression representative of forebrain, midbrain, and hindbrain development. Physiologic studies showed responses to glutamate and depolarization in many cells, consistent with neural behavior. The method of cerebral organoid generation described here facilitates access to this technology, enables scalable applications, and provides a potential pathway to translational applications where defined components are desirable. Tissue organoids are a promising technology with many potential applications, such as pharmaceutical screens and development of in vitro disease models, particularly for human polygenic conditions where animal models are insufficient. This work describes a robust and simple method for generating cerebral organoids from human induced pluripotent stem cells by using a chemically defined hydrogel material and chemically defined culture medium. This method, by virtue of its simplicity and use of defined materials, greatly facilitates access to cerebral organoid technology, enables scalable applications, and provides a potential pathway to translational applications where defined components are desirable. ©AlphaMed Press.

The protective effect of DNA on the rat cell membrane damage induced by ultraviolet radiation

International Nuclear Information System (INIS)

Ma Shouxiang; Zhong Jinyan

1988-01-01

The protective effect of DNA on the cell membrane damage induced by ultra-violet radiation was studied. Rat erythrocytes were used as experimental materials. Blood samples were taken from the rat, and centrifuged to separate the plasma. The cells were washed twice with isotonic saline, resuspended in normal saline solution and then irradiated by ultra-violet radiation. The DNA was added before or after irradiation. THe cell suspensions were kept at 5 deg C for 20 hours after irradiation, and then centrifuged. The supernatants were used for hemoglobin determination. The main results obtained may summarized as follows: the cell suspension of erythrocytes were irradiated for 5, 10 and 20 min. The amount of hemolysis induced by irradiation dosage revealed a direct proportional relationship. If DNA (20-40μg/ml) was applied before irradiation, the amount of hemolysis induced apparently decreased. The differences between the control and DNA treated were statistically significant, P<0.01, but insignificant for DNA added after irradiation

Generation of a restriction minus enteropathogenic Escherichia coli E2348/69 strain that is efficiently transformed with large, low copy plasmids

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Ward Jordan D

2008-08-01

Full Text Available Abstract Background Many microbes possess restriction-modification systems that protect them from parasitic DNA molecules. Unfortunately, the presence of a restriction-modification system in a given microbe also hampers genetic analysis. Although plasmids can be successfully conjugated into the enteropathogenic Escherichia coli strain E2348/69 and optimized protocols for competent cell preparation have been developed, we found that a large, low copy (~15 bioluminescent reporter plasmid, pJW15, that we modified for use in EPEC, was exceedingly difficult to transform into E2348/69. We reasoned that a restriction-modification system could be responsible for the low transformation efficiency of E2348/69 and sought to identify and inactivate the responsible gene(s, with the goal of creating an easily transformable strain of EPEC that could complement existing protocols for genetic manipulation of this important pathogen. Results Using bioinformatics, we identified genes in the unfinished enteropathogenic Escherichia coli (EPEC strain E2348/69 genome whose predicted products bear homology to the HsdM methyltransferases, HsdS specificity subunits, and HsdR restriction endonucleases of type I restriction-modification systems. We constructed a strain carrying a deletion of the conserved enzymatic domain of the EPEC HsdR homologue, NH4, and showed that its transformation efficiency was up to four orders of magnitude higher than that of the parent strain. Further, the modification capacity of NH4 remained intact, since plasmids that were normally recalcitrant to transformation into E2348/69 could be transformed upon passage through NH4. NH4 was unaffected in virulence factor production, since bundle forming pilus (BFP subunits and type III secreted (T3S proteins were present at equivalent levels to those seen in E2348/69. Further, NH4 was indistinguishable from E2348/69 in tissue culture infection model assays of localized adherence and T3S. Conclusion We

On the changes in optical interferometry induced by the relativistic motion of an optical medium

International Nuclear Information System (INIS)

Rakshit, Supantho

2016-01-01

We investigate the effects of the relativistic movement of an optical medium on the conditions of constructive and destructive interference, reflection and transmission pattern, and performance of spectroscopes. First, we consider the case of two beams reflected from a relativistically moving thin film and derive the conditions of constructive and destructive interference. Then we broaden the idea to multiple beam interference and formulate a new modified equation of reflection and transmission pattern of light from a relativistically moving plane parallel dielectric film. Finally, we determine the new effective resolving power of a Fabry–Perot spectroscope, which has a moving dielectric medium in its etalon. Here, we consider the case where the optical media move parallel to the plane of incidence of light. Throughout this paper, we use the basic Lorentz transformation of space and time co-ordinates and electromagnetic fields to conduct these investigations. This paper can strongly motivate undergraduate physics students to associate the concepts of special relativity and optical interference. (paper)

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

Science.gov (United States)

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

2012-05-01

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

Effect of oxidative stress induced by Brevibacterium sp. BS01 on a HAB causing species--Alexandrium tamarense.

Directory of Open Access Journals (Sweden)

Huajun Zhang

Full Text Available Harmful algal blooms occur all over the world, destroying aquatic ecosystems and threatening other organisms. The culture supernatant of the marine algicidal actinomycete BS01 was able to lysis dinoflagellate Alexandrium tamarense ATGD98-006. Physiological and biochemical responses to oxidative stress in A. tamarense were investigated to elucidate the mechanism involved in BS01 inhibition of algal growth. Transmission electron microscope analysis revealed that there were some chloroplast abnormalities in response to BS01 supernatant. The decrease in cellular-soluble protein content suggested that cell growth was greatly inhibited at high concentration of BS01 supernatant. The increase in the levels of reactive oxygen species (ROS and malondialdehyde contents following exposure to BS01 supernatant indicated that algal cells suffered from oxidative damage. The content of pigment was significantly decreased after 12 h treatment, which indicated that the accumulation of ROS destroyed pigment synthesis. Moreover, the decrease of Fv/Fm ratio suggested that in the photosynthetic system, the dominant sites producing ROS were destroyed by the supernatant of the BS01 culture. The activities of the antioxidant enzymes including superoxide dismutase and peroxidase increased in a short time and decreased slightly with increasing exposure time. A real-time PCR assay showed changes in the transcript abundances of two photosynthetic genes, psbA and psbD. The results showed that BS01 supernatant reduced the expression of the psbA gene after 2 h exposure, but the expression of the psbD gene was increased at concentrations of 1.0 and 1.5%. Our results demonstrated that the expression of the psbA gene was inhibited by the BS01 supernatant, which might block the electron transport chain, significantly enhancing ROS level and excess activity of the antioxidant system. The accumulation of ROS destoryed pigment synthesis and membrane integrity, and inhibited or

Plant Explants Grown on Medium Supplemented with Fe3O4 Nanoparticles Have a Significant Increase in Embryogenesis

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Inese Kokina

2017-01-01

Full Text Available Development of nanotechnology leads to the increasing release of nanoparticles in the environment that results in accumulation of different NPs in living organisms including plants. This can lead to serious changes in plant cultures which leads to genotoxicity. The aims of the present study were to detect if iron oxide NPs pass through the flax cell wall, to compare callus morphology, and to estimate the genotoxicity in Linum usitatissimum L. callus cultures induced by different concentrations of Fe3O4 nanoparticles. Two parallel experiments were performed: experiment A, where flax explants were grown on medium supplemented with 0.5 mg/l, 1 mg/l, and 1.5 mg/l Fe3O4 NPs for callus culture obtaining, and experiment B, where calluses obtained from basal MS medium were transported into medium supplemented with concentrations of NPs identical to experiment A. Obtained results demonstrate similarly in both experiments that 25 nm Fe3O4 NPs pass into callus cells and induce low toxicity level in the callus cultures. Nevertheless, calluses from experiment A showed 100% embryogenesis in comparison with experiment B where 100% rhizogenesis was noticed. It could be associated with different stress levels and adaptation time for explants and calluses that were transported into medium with Fe3O4 NPs supplementation.

The effect of bystander medium on the apoptotic process in HPV-G cells

International Nuclear Information System (INIS)

Maguire, P.; Lyng, F.; Seymour, C.; Mothersill, C.

2003-01-01

Full text: It has been shown in recent years that in both in vivo and in vitro situations irradiated cells cause what is known as the bystander effect. This presently unknown factor causes chromosomal aberrations, initiation of apoptosis and reduced clonogenic survival. Using the medium transfer method to study the bystander effect, this study investigated early events in the apoptotic cascade, which leads to cell death in cells receiving medium from irradiated cells but which were not themselves irradiated. Medium from irradiated ( 0.005Gy to 5Gy) human HPV G keratinocytes was harvested one hour after irradiation, sterile filtered and transferred on to unirradiated HPV-G cells. The appearance of apoptotic markers in the apoptotic cascade was monitored over a period of 48 hours following medium transfer. These apoptotic markers include loss of mitochondrial membrane potential, cytochrome c release and the activity of the death inducing caspase 3. Clonogenic survival of HPV-G cells over a nine day period was also monitored to assess the final survival of the cells. A TUNEL assay, which indicated the level of apoptosis over a 72 hour period after exposure to bystander medium was also performed. Data collected in this study indicates that for very low doses (0.005Gy) the appearance of well-characterised early 'apoptotic' markers such as changes in mitochondrial membrane potential doesn't mean the cell has committed to the apoptotic cascade leading to cell death. This has been illustrated for bystander medium from 0.005Gy irradiated cells, which causes mitochondrial membrane potential depolarisation after six-hour exposure but little difference has been noted for clonogenic survival for exposure to 0.005Gy bystander medium from that of the control. The results may help clarify how cells sector to death or survival following receipt of a signal from a radiation event

Antithymocyte Globulin Induces a Tolerogenic Phenotype in Human Dendritic Cells

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Tobias Roider

2016-12-01

Full Text Available Antithymocyte globulin (ATG is used in the prevention of graft-versus-host disease during allogeneic hematopoietic stem cell transplantation. It is generally accepted that ATG mediates its immunosuppressive effect primarily via depletion of T cells. Here, we analyzed the impact of ATG-Fresenius (now Grafalon® on human monocyte-derived dendritic cells (DC. ATG induced a semi-mature phenotype in DC with significantly reduced expression of CD14, increased expression of HLA-DR, and intermediate expression of CD54, CD80, CD83, and CD86. ATG-DC showed an increase in IL-10 secretion but no IL-12 production. In line with this tolerogenic phenotype, ATG caused a significant induction of indoleamine 2,3-dioxygenase expression and a concomitant increase in levels of tryptophan metabolites in the supernatants of DC. Further, ATG-DC did not induce the proliferation of allogeneic T cells in a mixed lymphocyte reaction but actively suppressed the T cell proliferation induced by mature DC. These data suggest that besides its well-known effect on T cells, ATG modulates the phenotype of DC in a tolerogenic way, which might constitute an essential part of its immunosuppressive action in vivo.

Aspiration of barium contrast medium in an elderly man with disordered swallowing

OpenAIRE

Bağcı Ceyhan, B.; Çelikel, T.; Koç, M.; Ahıskalı, R.; Biren, T.; Ataizi Çelikel, Ç.

1995-01-01

The aspiration of contrast medium during the investigation of gastrointestinal diseases is a well recognized hazard, particularly in patients with swallowing disorders. A case is reported in which accidental aspiration of contrast barium occurred owing to disordered swallowing in an elderly man. The infiltration on chest x-ray persisted 2 years after barium contrast aspiration. Inflammatory reaction and retractile, granular material observed in lung biopsy specimens suggested barium-induced p...

Virulent and avirulent strains of equine arteritis virus induce different quantities of TNF-α and other proinflammatory cytokines in alveolar and blood-derived equine macrophages

International Nuclear Information System (INIS)

Moore, Brian D.; Balasuriya, Udeni B.R.; Watson, Johanna L.; Bosio, Catharine M.; MacKay, Robert J.; MacLachlan, N. James

2003-01-01

Equine arteritis virus (EAV) infects endothelial cells (ECs) and macrophages in horses, and many of the clinical manifestations of equine viral arteritis (EVA) reflect vascular injury. To further evaluate the potential role of EAV-induced, macrophage-derived cytokines in the pathogenesis of EVA, we infected cultured equine alveolar macrophages (AMphi), blood monocyte-derived macrophages (BMphi), and pulmonary artery ECs with either a virulent (KY84) or an avirulent (CA95) strain of EAV. EAV infection of equine AMphi, BMphi, and ECs resulted in their activation with increased transcription of genes encoding proinflammatory mediators, including interleukin (IL)-1β, IL-6, IL-8, and tumor necrosis factor (TNF)-α. Furthermore, the virulent KY84 strain of EAV induced significantly higher levels of mRNA encoding proinflammatory cytokines in infected AMphi and BMphi than did the avirulent CA95 strain. Treatment of equine ECs with the culture supernatants of EAV-infected AMphi and BMphi also resulted in EC activation with cell surface expression of E-selectin, whereas infection of ECs with purified EAV alone caused only minimal expression of E-selectin. The presence of TNF-α in the culture supernatants of EAV-infected equine AMphi, BMphi, and ECs was confirmed by bioassay, and the virulent KY84 strain of EAV induced significantly more TNF-α in all cell types than did the avirulent CA95 strain. Thus, the data indicate that EAV-induced, macrophage-derived cytokines may contribute to the pathogenesis of EVA in horses, and that the magnitude of the cytokine response of equine AMphi, BMphi, and ECs to EAV infection reflects the virulence of the infecting virus strain

Tattoo: a multifaceted medium of communication

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Christian Wymann

2010-11-01

Full Text Available This article suggests the systems theoretical distinction of form/medium as a useful tool for distinguishing social phenomena that might look as if they stem from the same process. This is shown to be the case for the tattoo and tattooing. The tattoo is conceived as a medium of communication through which different forms of communication emerge. Tattooing is one of these forms of communication that shapes the medium in a particular way. The current article sheds a special light on its intricate, communicational constellation, for which the concept of parallax is suggested. Law, medicine and cosmetics as other forms of communication use the medium of tattoo in their own way as well. The form/medium distinction allows us to grasp these different forms of communication, while it shows that they share the tattoo as medium. The article’s ultimate goal is to illustrate that the tattoo figures as a multifaceted medium of communication.

Purification and characterisation of Cyclodextrin glycosyltransferase ...

African Journals Online (AJOL)

EXPER

2012-06-05

Jun 5, 2012 ... require higher temperatures for the liquefaction of starch at which CGTases ... growth medium by centrifugation at 12000 x g for 15 min. Cell-free supernatant ..... 95°C, thermal inactivation was only 13% (Figure 3). Enzyme ...

Medium-term dynamics of a middle Adriatic barred beach

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M. Postacchini

2017-09-01

Full Text Available In recent years, attention has been paid to beach protection by means of soft and hard defenses. Along the Italian coast of the Adriatic Sea, sandy beaches are the most common landscape feature and around 70 % of the Marche region's coast (central Adriatic is protected by defense structures. The longest free-from-obstacle nearshore area in the region includes the beach of Senigallia, frequently monitored in the last decades and characterized by a multiple bar system, which represents a natural beach defense. The bathymetries surveyed in 2006, 2010, 2011, 2012 and 2013 show long-term stability, confirmed by a good adaptation of an analyzed stretch of the beach to the Dean-type equilibrium profile, though a strong short- to medium-term variability of the wave climate has been observed during the monitored periods. The medium-term dynamics of the beach, which deal with the evolution of submerged bars and are of the order of years or seasons, have been related to the wave climate collected, during the analyzed temporal windows, by a wave buoy located about 40 km off Senigallia. An overall interpretation of the hydrodynamics, sediment characteristics and seabed morphology suggests that the wave climate is fundamental for the morphodynamic changes of the beach in the medium term. These medium-term time ranges during which waves mainly come from NNE/ESE are characterized by a larger/smaller steepness and by a larger/smaller relative wave height, and seem to induce seaward/shoreward bar migration as well as bar smoothing/steepening. Moving southeastward, the bar dimension increases, while the equilibrium profile shape suggests the adaptation to a decreasing sediment size in the submerged beach. This is probably due to the presence of both the harbor jetty and river mouth north of the investigated area.

Genistein suppresses Prevotella intermedia lipopolysaccharide-induced inflammatory response in macrophages and attenuates alveolar bone loss in ligature-induced periodontitis.

Science.gov (United States)

Choi, Eun-Young; Bae, Seung Han; Ha, Min Hee; Choe, So-Hui; Hyeon, Jin-Yi; Choi, Jeom-Il; Choi, In Soon; Kim, Sung-Jo

2016-02-01

Genistein is a major isoflavone subclass of flavonoids found in soybean and a potent tyrosine kinase inhibitor. The present study aimed to assess the effect of genistein on the production of proinflammatory mediators in murine macrophages stimulated with lipopolysaccharide (LPS) isolated from Prevotella intermedia, a pathogen associated with different forms of periodontal disease, and to evaluate its possible influence on alveolar bone loss in ligature-induced periodontitis using micro-computed tomography (micro-CT) analysis as well. LPS was isolated from P. intermedia ATCC 25611 by using the standard hot phenol-water method. Culture supernatants were analyzed for nitric oxide (NO) and interleukin-6 (IL-6). Inducible NO synthase (iNOS) protein expression was evaluated by immunoblot analysis. Real-time PCR was carried out to measure iNOS and IL-6 mRNA expression. In addition, effect of genistein on alveolar bone loss was evaluated in a rat model of experimental periodontitis using micro-CT analysis. Genistein significantly attenuated P. intermedia LPS-induced production of iNOS-derived NO and IL-6 with attendant decrease in their mRNA expression in RAW264.7 cells. In addition, when genistein was administered to rats, decreases in alveolar bone height and bone volume fraction induced by ligature placement were significantly inhibited. Genistein administration also prevented ligature-induced alterations in the microstructural parameters of trabecular bone, including trabecular thickness, trabecular separation, bone mineral density and structure model index. While additional studies are required, we suggest that genistein could be utilized for the therapy of human periodontitis in the future. Copyright © 2015 Elsevier Ltd. All rights reserved.

Rain-induced bistatic scattering at 60 GHz

NARCIS (Netherlands)

Zanden, van der H.T.; Watson, R.J.; Herben, M.H.A.J.

2007-01-01

This paper presents the results of a study into the modeling and prediction of rain-induced bistatic scattering at 60 GHz. The bistatic radar equation together withMie theory is applied as the basis for calculating the scattering. Together with the attenuation induced by the medium before and after

The assessment of cytokines in Quantiferon supernatants for the diagnosis of latent TB infection in a tribal population of Melghat, India.

Science.gov (United States)

Bapat, Prachi R; Husain, Aliabbas A; Daginawala, Hatim F; Agrawal, Neha P; Panchbhai, Milind S; Satav, Ashish R; Taori, Girdhar M; Kashyap, Rajpal S

2015-01-01

The tuberculin skin test (TST) and interferon-gamma release assays (IGRA), namely, the QuantiFERON-TB Gold test (QFT), remain the standard immunological diagnostic tools for latent tuberculosis (TB) infection (LTBI). However, the sub-optimal detection rates of both of these tests are major impediments in recognizing the population at risk. This study was aimed at evaluating additional cytokines besides interferon-gamma (IFN-γ) as biomarkers for improving LTBI diagnosis in the tribal population of Melghat, India. Seventy-four close TB contacts were stratified by QFT and TST results into: (i) QFT+/TST+ (n = 26), (ii) QFT+/TST- (n = 12), (iii) QFT-/TST- (n = 35) and (iv) QFT-/TST+ (n = 1) groups. A panel of cytokines (IL-6, IL-10, TNF-α and IL-2R) was then evaluated in antigen-stimulated QFT cell-free culture supernatants using IMMULITE-1000, an automated immunoassay analyzer. Cytokine estimation showed significantly higher levels of IL-6 in the QFT+/TST+ group, while significantly higher levels of IL-10 were found in the QFT-/TST- group. Correlation analysis identified a positive correlation between IL-6 and the QFT response (r = 0.6723, P < 0.0001), while a negative correlation was seen between QFT and IL-10 expression (r = -0.3271, P = 0.0044). Similarly, IL-6 was positively correlated with TST levels (r = 0.6631, P <0 .0001), and conversely, a negative correlation was found between TST and IL-10 expression (r = -0.5698, P < 0.0001). The positive and negative predictive values of IL-6 were found to be 92.59 and 93.33%, respectively, and the positive and negative predictive values of IL-10 were 96.55 and 91.18%, respectively. No significant impact of the demographic characteristics on cytokine positivity was observed. Our preliminary results suggest that the evaluation of additional cytokines in QFT cell-free culture supernatants may be valuable for the identification of LTBI. Combining IL-6 and IL-10 with QFT and/or TST could markedly improve the detection

Characterisation of STEC and other diarrheic E. coli isolated on CHROMagar™STEC at a tertiary referral hospital, Cape Town.

Science.gov (United States)

Kalule, John Bosco; Keddy, Karen H; Nicol, Mark P

2018-06-08

Shiga toxin producing E. coli (STEC) is an emerging zoonotic pathogen that can cause acute renal failure, especially in children. Clinical microbiology laboratories may fail to detect STEC and other diarrhoeic E. coli unless purposive rigorous screening procedures are followed using appropriate diagnostic technology; CHROMagar™STEC has rarely been used for isolation of African diarrhoeic E. coli hence characteristics of isolates on this medium are not yet fully understood. This study aimed to determine the prevalence and characteristics of STEC and other diarrhoeic E. coli isolated on CHROMagar™STEC from stool samples submitted to the microbiology laboratory of a South African public sector tertiary care hospital. In total, 733 stool samples were tested. Of these, 4.5% (33/733) possessed diarrhoeic E. coli. Of the diarrheic E. coli, 5/33 (15.2%) were STEC, 15/33 (45.5%) EAggEC, 6/33 (18.2%) atypical EPEC, 5/33 (15.2%) typical EPEC, and 1/33 (3%) DAEC. None of the STEC isolates had been identified by routine testing (based on using sorbitol media to test for E. coli O157: H7 strains and not the other STEC) in the laboratory. Of the 33 strains, 55% (95% CI = 40.8-72.7) showed resistance to ampicillin. CHROMagar™STEC enabled detection of tellurite - resistant diarrhoeic E. coli that would be missed using routine methods. Further studies are needed to determine the proportion and characteristics of those which might have been missed using this approach.

Fermented dairy products modulate Citrobacter rodentium-induced colonic hyperplasia.

Science.gov (United States)

Collins, James W; Chervaux, Christian; Raymond, Benoit; Derrien, Muriel; Brazeilles, Rémi; Kosta, Artemis; Chambaud, Isabelle; Crepin, Valerie F; Frankel, Gad

2014-10-01

We evaluated the protective effects of fermented dairy products (FDPs) in an infection model, using the mouse pathogen Citrobacter rodentium (CR). Treatment of mice with FDP formulas A, B, and C or a control product did not affect CR colonization, organ specificity, or attaching and effacing lesion formation. Fermented dairy product A (FDP-A), but neither the supernatant from FDP-A nor β-irradiated (IR) FDP-A, caused a significant reduction in colonic crypt hyperplasia and CR-associated pathology. Profiling the gut microbiota revealed that IR-FDP-A promoted higher levels of phylotypes belonging to Alcaligenaceae and a decrease in Lachnospiraceae (Ruminococcus) during CR infection. Conversely, FDP-A prevented a decrease in Ruminococcus and increased Turicibacteraceae (Turicibacter). Importantly, loss of Ruminococcus and Turicibacter has been associated with susceptibility to dextran sodium sulfate-induced colitis. Our results demonstrate that viable bacteria in FDP-A reduced CR-induced colonic crypt hyperplasia and prevented the loss of key bacterial genera that may contribute to disease pathology. © The Author 2014. Published by Oxford University Press on behalf of the Infectious Diseases Society of America.

A novel cell growth-promoting factor identified in a B cell leukemia cell line, BALL-1

International Nuclear Information System (INIS)

Dao, T.; Holan, V.; Minowada, J.

1993-01-01

A novel leukemia cell growth-promoting activity has been identified in the culture supernatant from a human B cell leukemia cell line, BALL-1. The supernatant from unstimulated cultures of the BALL-1 cells significantly promoted the growth of 16 out of 24 leukemia/lymphoma cell lines of different lineages (T, B and non-lymphoid) in a minimal concentration of fetal bovine serum (FBS), and 5 out of 12 cases of fresh leukemia cells in FBS-free medium. The growth-promoting sieve filtration and dialysis. The MW of the factor was less than 10 kDa. The growth-promoting activity was heat and acid stable and resistant to trypsin treatment. The factor isolated from the BALL-1 supernatant was distinct from known polypeptide growth factors with MW below 10 kDa, such as epidermal growth factor, transforming growth factor α, insulin-like growth factor I (IGF-I), IGF-II and insulin, as determine by specific antibodies and by cell-growth-promoting tests. The factor is the BALL-1 supernatant did not promote the proliferation of normal human fresh peripheral blood lymphocytes or mouse fibroblast cell line, BALB/C 3T3. In addition to the BALL-1 supernatant, a similar growth-promoting activity was found in the culture supernatant from 13 of 17 leukemia/lymphoma cell lines tested. The activity in these culture supernatant promoted the growth of leukemia/lymphoma cell lines in autocrine and/or paracrine fashions. These observations suggest that the low MW cell growth-promoting activity found in the BALL-1 culture supernatant is mediated by a novel factor which may be responsible for the clonal expansion of particular leukemic clones. (author)

Transepithelial activation of human leukocytes by probiotics and commensal bacteria: role of Enterobacteriaceae-type endotoxin

DEFF Research Database (Denmark)

Bäuerlein, A.; Ackermann, S.; Parlesak, Alexandr

2009-01-01

The goal of the current study was to clarify whether commercially available probiotics induce greater trans-epithelial activation of human leukocytes than do commensal, food-derived and pathogenic bacteria and to identify the compounds responsible for this activation. Eleven different bacterial...... Escherichia coli K12, probiotic E. coli Nissle, EPEC) induced basolateral production of TNF-alpha, IFN-gamma, IL 6, 8, and 10. Gram-positive probiotics (Lactobacillus spp. and Bifidobacterium spp.) had virtually no effect. In addition, commensals (Enterococcus faecalis, Bacteroides vulgatus) and food...... (polymyxin, colistin) completely abrogated transepithelial activation of leukocytes. Enterobacteriaceae-type endotoxin is a crucial factor in transepithelial stimulation of leukocytes, regardless of whether it is produced by probiotics or other bacteria. Hence, transepithelial stimulation ofleukocytes...

Medium-scale traveling ionospheric disturbances triggered by Super Typhoon Nepartak (2016)

Science.gov (United States)

Chou, M. Y.; Lin, C. C. H.; Yue, J.; Chang, L. C.; Tsai, H. F.; Chen, C. H.

2017-12-01

Two remarkable typhoon-induced traveling ionospheric disturbances (TIDs) with concentric andnorthwest-southeast (NW-SE) alignments, respectively, associated with concentric gravity waves (CGWs) andionospheric instabilities possibly seeded by CGWs, were observed in total electron content (TEC) derived fromground-based Global Navigation Satellite System networks in Taiwan and Japan when the Category 5 SuperTyphoon Nepartak approached Taiwan on 7 July 2016. The concentric TIDs (CTIDs) first appear withhorizontal phase velocities of 161-200 m/s, horizontal wavelengths of 160-270 km, and periods of 15-22 min during 08:00-11:20 UT. Following the CTIDs, the NW-SE aligned nighttime medium-scale TIDs(MSTIDs) are formed on the west edge of the CTIDs over the Taiwan Strait during 11:30-14:00 UT. It issuggested that the MSTIDs are produced by the electrodynamical coupling of Perkins instability andCGW-induced polarization electric fields. This study proposes connections of typhoon-induced CTIDs andsubsequently occurring MSTIDs in the low-latitude ionosphere.

Chemical inducible promotor used to obtain transgenic plants with a silent marker

Science.gov (United States)

Chua, Nam-Hai; Aoyama, Takashi

2000-01-01

A chemically inducible promoter is described which may be used to transform plants with genes which are easily regulatable by adding plants or plant cells to a medium containing an inducer of the promoter or by removing the plants or plant cells from such medium. The promoter described is one which is inducible by a glucocorticoid which is not endogenous to plants. Such promoters may be used with a variety of genes such as ipt or knotted1 to induce shoot formation in the presence of a glucocorticoid. The promoter may also be used with antibiotic or herbicide resistance genes which are then regulatable by the presence or absence of inducer rather than being constitutive. Other examples of genes which may be placed under the control of the inducible promoter are also presented.

Effective medium theory for anisotropic metamaterials

KAUST Repository

Zhang, Xiujuan

2015-01-20

Materials with anisotropic material parameters can be utilized to fabricate many fascinating devices, such as hyperlenses, metasolids, and one-way waveguides. In this study, we analyze the effects of geometric anisotropy on a two-dimensional metamaterial composed of a rectangular array of elliptic cylinders and derive an effective medium theory for such a metamaterial. We find that it is possible to obtain a closed-form analytical solution for the anisotropic effective medium parameters, provided the aspect ratio of the lattice and the eccentricity of the elliptic cylinder satisfy certain conditions. The derived effective medium theory not only recovers the well-known Maxwell-Garnett results in the quasi-static regime, but is also valid beyond the long-wavelength limit, where the wavelength in the host medium is comparable to the size of the lattice so that previous anisotropic effective medium theories fail. Such an advance greatly broadens the applicable realm of the effective medium theory and introduces many possibilities in the design of structures with desired anisotropic material characteristics. A real sample of a recently theoretically proposed anisotropic medium, with a near-zero index to control the flux, is achieved using the derived effective medium theory, and control of the electromagnetic waves in the sample is clearly demonstrated.

The Spread of a Noise Field in a Dispersive Medium

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Cohen Leon

2010-01-01

Full Text Available We discuss the production of induced noise by a pulse and the propagation of the noise in a dispersive medium. We present a simple model where the noise is the sum of pulses and where the mean of each pulse is random. We obtain explicit expressions for the standard deviation of the spatial noise as a function of time. We also formulate the problem in terms of a time-frequency phase space approach and in particular we use the Wigner distribution to define the spatial/spatial-frequency distribution.

Simultaneous determination and stability studies of linezolid, meropenem and vancomycin in bacterial growth medium by high-performance liquid chromatography.

Science.gov (United States)

Wicha, Sebastian G; Kloft, Charlotte

2016-08-15

For pharmacokinetic/pharmacodynamic (PK/PD) assessment of antibiotics combinations in in vitro infection models, accurate and precise quantification of drug concentrations in bacterial growth medium is crucial for derivation of valid PK/PD relationships. We aimed to (i) develop a high-performance liquid chromatography (HPLC) assay to simultaneously quantify linezolid (LZD), vancomycin (VAN) and meropenem (MER), as typical components of broad-spectrum antibiotic combination therapy, in bacterial growth medium cation-adjusted Mueller-Hinton broth (CaMHB) and (ii) determine the stability profiles of LZD, VAN and MER under conditions in in vitro infection models. To separate sample matrix components, the final method comprised the pretreatment of 100μL sample with 400μL methanol, the evaporation of supernatant and its reconstitution in water. A low sample volume of 2μL processed sample was injected onto an Accucore C-18 column (2.6μm, 100×2.1mm) coupled to a Dionex Ultimate 3000 HPLC+ system. UV detection at 251, 240 and 302nm allowed quantification limits of 0.5, 2 and 0.5μg/mL for LZD, VAN and MER, respectively. The assay was successfully validated according to the relevant EMA guideline. The rapid method (14min) was successfully applied to quantify significant degradation of LZD, VAN and MER in in vitro infection models: LZD was stable, VAN degraded to 90.6% and MER to 62.9% within 24h compared to t=0 in CaMHB at 37°C, which should be considered when deriving PK/PD relationships in in vitro infection models. Inclusion of further antibiotics into the flexible gradient-based HPLC assay seems promising. Copyright © 2016 Elsevier B.V. All rights reserved.

Molecular Characterization of the Elaeis guineensis Medium-Chain Fatty Acid Diacylglycerol Acyltransferase DGAT1-1 by Heterologous Expression in Yarrowia lipolytica.

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Laure Aymé

Full Text Available Diacylglycerol acyltransferases (DGAT are involved in the acylation of sn-1,2-diacylglycerol. Palm kernel oil, extracted from Elaeis guineensis (oil palm seeds, has a high content of medium-chain fatty acids mainly lauric acid (C12:0. A putative E. guineensis diacylglycerol acyltransferase gene (EgDGAT1-1 is expressed at the onset of lauric acid accumulation in the seed endosperm suggesting that it is a determinant of medium-chain triacylglycerol storage. To test this hypothesis, we thoroughly characterized EgDGAT1-1 activity through functional complementation of a Yarrowia lipolytica mutant strain devoid of neutral lipids. EgDGAT1-1 expression is sufficient to restore triacylglycerol accumulation in neosynthesized lipid droplets. A comparative functional study with Arabidopsis thaliana DGAT1 highlighted contrasting substrate specificities when the recombinant yeast was cultured in lauric acid supplemented medium. The EgDGAT1-1 expressing strain preferentially accumulated medium-chain triacylglycerols whereas AtDGAT1 expression induced long-chain triacylglycerol storage in Y. lipolytica. EgDGAT1-1 localized to the endoplasmic reticulum where TAG biosynthesis takes place. Reestablishing neutral lipid accumulation in the Y. lipolytica mutant strain did not induce major reorganization of the yeast microsomal proteome. Overall, our findings demonstrate that EgDGAT1-1 is an endoplasmic reticulum DGAT with preference for medium-chain fatty acid substrates, in line with its physiological role in palm kernel. The characterized EgDGAT1-1 could be used to promote medium-chain triacylglycerol accumulation in microbial-produced oil for industrial chemicals and cosmetics.

Fibrinogen-Induced Streptococcus mutans Biofilm Formation and Adherence to Endothelial Cells

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Telma Blanca Lombardo Bedran

2013-01-01

Full Text Available Streptococcus mutans, the predominant bacterial species associated with dental caries, can enter the bloodstream and cause infective endocarditis. The aim of this study was to investigate S. mutans biofilm formation and adherence to endothelial cells induced by human fibrinogen. The putative mechanism by which biofilm formation is induced as well as the impact of fibrinogen on S. mutans resistance to penicillin was also evaluated. Bovine plasma dose dependently induced biofilm formation by S. mutans. Of the various plasma proteins tested, only fibrinogen promoted the formation of biofilm in a dose-dependent manner. Scanning electron microscopy observations revealed the presence of complex aggregates of bacterial cells firmly attached to the polystyrene support. S. mutans in biofilms induced by the presence of fibrinogen was markedly resistant to the bactericidal effect of penicillin. Fibrinogen also significantly increased the adherence of S. mutans to endothelial cells. Neither S. mutans cells nor culture supernatants converted fibrinogen into fibrin. However, fibrinogen is specifically bound to the cell surface of S. mutans and may act as a bridging molecule to mediate biofilm formation. In conclusion, our study identified a new mechanism promoting S. mutans biofilm formation and adherence to endothelial cells which may contribute to infective endocarditis.

Induction of chagasic-like arrhythmias in the isolated beating hearts of healthy rats perfused with Trypanosoma cruzi-conditioned medium

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H. Rodriguez-Angulo

2013-01-01

Full Text Available Chagas' myocardiopathy, caused by the intracellular protozoan Trypanosoma cruzi, is characterized by microvascular alterations, heart failure and arrhythmias. Ischemia and arrythmogenesis have been attributed to proteins shed by the parasite, although this has not been fully demonstrated. The aim of the present investigation was to study the effect of substances shed by T. cruzi on ischemia/reperfusion-induced arrhythmias. We performed a triple ischemia-reperfusion (I/R protocol whereby the isolated beating rat hearts were perfused with either Vero-control or Vero T. cruzi-infected conditioned medium during the different stages of ischemia and subsequently reperfused with Tyrode's solution. ECG and heart rate were recorded during the entire experiment. We observed that triple I/R-induced bradycardia was associated with the generation of auricular-ventricular blockade during ischemia and non-sustained nodal and ventricular tachycardia during reperfusion. Interestingly, perfusion with Vero-infected medium produced a delay in the reperfusion-induced recovery of heart rate, increased the frequency of tachycardic events and induced ventricular fibrillation. These results suggest that the presence of parasite-shed substances in conditioned media enhances the arrhythmogenic effects that occur during the I/R protocol.

Two-gluon emission and interference in a thin QCD medium: insights into jet formation

International Nuclear Information System (INIS)

Casalderrey-Solana, Jorge; Pablos, Daniel; Tywoniuk, Konrad

2016-01-01

In heavy-ion collisions, an abundant production of high-energy QCD jets allows to study how these multiparticle sprays are modified as they pass through the quark-gluon plasma. In order to shed new light on this process, we compute the inclusive two-gluon rate off a hard quark propagating through a color deconfined medium at first order in medium opacity. We explicitly impose an energy ordering of the two emitted gluons, such that the “hard” gluon can be thought of as belonging to the jet substructure while the other is a “soft” emission (which can be collinear or medium-induced). Our analysis focusses on two specific limits that clarify the modification of the additional angle- and formation time-ordering of splittings. In one limit, the formation time of the “hard” gluon is short compared to the “soft” gluon formation time, leading to a probabilistic formula for production of and subsequent radiation off a quark-gluon antenna. In the other limit, the ordering of formation is reverted, which automatically leads to the fact that the jet substructure is resolved by the medium. We observe in this case a characteristic delay: the jet radiates as one color current (quark) up to the formation of the “hard” gluon, at which point we observe the onset of radiation of the new color current (gluon). Within our kinematic constraints, our computation supports a picture in which the in-medium jet dynamics are described as a collection of subsequent antennas which are resolved by the medium according to their transverse extent.

Enhancement of excision-repair efficiency by conditioned medium from density-inhibited cultures in V79 Chinese hamster cells

International Nuclear Information System (INIS)

Nakano, S.

1979-01-01

Conditioned medium from density-inhibited V79 Chinese hamster cell cultures, given as a post-treatment to UV-irradiated homologous cells, was demonstrated to reduce the lethal action of ultraviolet light by temporarily blocking DNA replication. Since the increased survival was not affected by various nontoxic concentrations of caffeine, such protective effect would be attributable to the prolonged intervention of excision repair before DNA replication during the post-treatment period. The influence of conditioned medium on the UV-induced mutation at the ouabain-resistance locus was also examined and a significant decrease in mutation frequecy was noted. The observed reduction in killing and mutation as a result of post-incubation in conditioned medium, which delays DNA replication, would be interpreted as evidence that conditioned medium provides a longer period of time for an error-free excision-repair process, leaving lesion in DNA available for error-prone post-replication repair. (Auth.)

Isolation of Lactobacillus salivarius 1077 (NRRL B-50053) and characterization of its bacteriocin and spectra of antimicrobial activity

Science.gov (United States)

Lactobacillus salivarius 1077 (NRRL B-50053) was isolated from poultry intestinal materials after demonstrating in-vitro anti-Campylobacter jejuni activity. The isolate was then used for in-vitro fermentation. The protein content of the cell-free supernatant from the spent medium was precipitated ...

Lactobacillus salivarius 1077 (NRRL B-50053) bacteriocin

Science.gov (United States)

Lactobacillus salivarius 1077 (NRRL B-50053) was isolated from poultry intestinal materials after demonstrating in-vitro anti-Campylobacter jejuni activity. The isolate was then used for in-vitro fermentation. The protein content of the cell-free supernatant from the spent medium was precipitated ...

Interrelation between striction forces in dielectrics and optically induced forces in transparent media

International Nuclear Information System (INIS)

Torchigin, V P; Torchigin, A V

2012-01-01

Optically induced forces applied to a transparent optical medium, which is inserted in a closed plane optical resonator, are calculated by means of an analysis of the changes in the eigenfrequency and energy stored in the resonator at various positions of the medium. These forces are compared with striction forces applied to the medium considered as a dielectric placed in an alternate electrical field within the resonator. It is shown that the optically induced forces are equal to the striction forces. The results of using the classical formula for striction forces in electrostatics are considered. (paper)

The predictive value of the product of contrast medium volume and urinary albumin/creatinine ratio in contrast-induced acute kidney injury.

Science.gov (United States)

Wang, Chunrui; Ma, Shuai; Deng, Bo; Lu, Jianxin; Shen, Wei; Jin, Bo; Shi, Haiming; Ding, Feng

2017-11-01

Preexisting renal impairment and the amount of contrast media are the most important risk factors for contrast-induced acute kidney injury (CI-AKI). We aimed to investigate whether the product of contrast medium volume and urinary albumin/creatinine ratio (CMV × UACR) would be a better predictor of CI-AKI in patients undergoing nonemergency coronary interventions. This was a prospective single-center observational study, and 912 consecutive patients who were exposed to contrast media during coronary interventions were investigated prospectively. CI-AKI is defined as a 44.2 μmol/L rise in serum creatinine or a 25% increase, assessed within 48 h after administration of contrast media in the absence of other causes. Fifty patients (5.48%) developed CI-AKI. The urinary albumin/creatinine ratio (UACR) (OR = 1.002, 95% CI = 1.000-1.003, p = .012) and contrast medium volume (CMV) (OR = 1.008, 95% CI = 1.001-1.014, p = .017) were independent risk factors for the development of CI-AKI. The area under the ROC curve of CMV, UACR and CMV × UACR were 0.662 (95% CI = 0.584-0.741, p < .001), 0.761 (95% CI = 0.674-0.847, p < .001) and 0.808 (95% CI = 0.747-0.896, p < .001), respectively. The cutoff value of CMV × UACR to predict CI-AKI was 1186.2, with 80.0% sensitivity and 62.2% specificity. The product of CMV and UACR (CMV × UACR) might be a predictor of CI-AKI in patients undergoing nonemergency coronary interventions, which was superior to CMV or UACR alone.

Surface plasmon enhancement in gold nanoparticles in the presence of an optical gain medium: an analysis

Energy Technology Data Exchange (ETDEWEB)

Sathiyamoorthy, K; Sreekanth, K V; Sidharthan, R; Murukeshan, V M [School of Mechanical and Aerospace Engineering, Nanyang Technological University, 50 Nanyang Avenue, Singapore 639798 (Singapore); Xing Bengang, E-mail: mmurukeshan@ntu.edu.sg [Division of Chemistry and Biological Chemistry, School of Physical and Mathematical Sciences, Nanyang Technological University, 21 Nanyang Link, Singapore 637371 (Singapore)

2011-10-26

The localized surface plasmon (LSP) enhancement in a gold nanoparticle is demonstrated in this paper. The enhancement of LSP is influenced by both size and the dielectric gain medium surrounding the nanoparticles. The nanoparticle is found to induce plasmonic enhancement of varying degrees depending on its size, and it is inferred that a gold nanoparticle of size 60 nm exhibits the maximum LSP for 532 nm excitation. Singularity due to cancellation of SP loss by an infinite gain medium and LSP enhancement are studied using a pump-probe Rayleigh scattering experiment. Gold nanoparticles of average size 60 nm exhibit the lowest threshold power to observe Rayleigh scattering. Furthermore, compared with the bare nanoparticles, a 12.5 fold enhancement of LSP is observed when the nanoparticle of average size 60 nm is kept in the gain medium.

Activation of mGluR5 induces spike afterdepolarization and enhanced excitability in medium spiny neurons of the nucleus accumbens by modulating persistent Na+ currents

Science.gov (United States)

D’Ascenzo, Marcello; Podda, Maria Vittoria; Fellin, Tommaso; Azzena, Gian Battista; Haydon, Philip; Grassi, Claudio

2009-01-01

The involvement of metabotropic glutamate receptors type 5 (mGluR5) in drug-induced behaviours is well-established but limited information is available on their functional roles in addiction-relevant brain areas like the nucleus accumbens (NAc). This study demonstrates that pharmacological and synaptic activation of mGluR5 increases the spike discharge of medium spiny neurons (MSNs) in the NAc. This effect was associated with the appearance of a slow afterdepolarization (ADP) which, in voltage-clamp experiments, was recorded as a slowly inactivating inward current. Pharmacological studies showed that ADP was elicited by mGluR5 stimulation via G-protein-dependent activation of phospholipase C and elevation of intracellular Ca2+ levels. Both ADP and spike aftercurrents were significantly inhibited by the Na+ channel-blocker, tetrodotoxin (TTX). Moreover, the selective blockade of persistent Na+ currents (INaP), achieved by NAc slice pre-incubation with 20 nm TTX or 10 μm riluzole, significantly reduced the ADP amplitude, indicating that this type of Na+ current is responsible for the mGluR5-dependent ADP. mGluR5 activation also produced significant increases in INaP, and the pharmacological blockade of this current prevented the mGluR5-induced enhancement of spike discharge. Collectively, these data suggest that mGluR5 activation upregulates INaP in MSNs of the NAc, thereby inducing an ADP that results in enhanced MSN excitability. Activation of mGluR5 will significantly alter spike firing in MSNs in vivo, and this effect could be an important mechanism by which these receptors mediate certain aspects of drug-induced behaviours. PMID:19433572

Calculations of lightning-induced voltages in medium voltage distribution lines

Energy Technology Data Exchange (ETDEWEB)

Munhoz Rojas, Patricio E. [Instituto de Tecnologia para o Desenvolvimento (LACTEC/UFPR), Curitiba, PR (Brazil)], E-mail: patricio@lactec.org.br; Pinto, Cleverson Luiz da Silva [Companhia Paranaense de Energia (COPEL), Curitiba, PR (Brazil)], E-mail: cleverson@copel.com

2007-07-01

The objective of this paper is to show the results of a new computer program, written in Mat Lab environment, that is intended to calculate the lightning induced voltages in multi-conductor non-homogeneous lines, in order to be able to evaluate the impact of the usual protective measures implemented against lightning-induced disturbances. The main new features of this program are: a coupling model in terms of the scalar potentials referred to a remote ground was adopted; the coupling to the vertical conductors was considered in a manner similar to rest of the line; the describing equations were converted into a system of coupled ordinary differential equations, by a discretization only in space, which was subsequently solved using the powerful ODE solvers existing in Mat Lab. The validation of the new program has been performed by comparing its predicted results with other theoretical and computational results available in the literature and also with other experimental results published in the literature. It is shown that the agreement with other well established theoretical results is very good and, also, that the agreement between the newly calculated results and some published experimental results is better than previously obtained results. (author)

Ion-induced particle desorption in time-of-flight medium energy ion scattering

Science.gov (United States)

Lohmann, S.; Primetzhofer, D.

2018-05-01

Secondary ions emitted from solids upon ion impact are studied in a time-of-flight medium energy ion scattering (ToF-MEIS) set-up. In order to investigate characteristics of the emission processes and to evaluate the potential for surface and thin film analysis, experiments employing TiN and Al samples were conducted. The ejected ions exhibit a low initial kinetic energy of a few eV, thus, requiring a sufficiently high acceleration voltage for detection. Molecular and atomic ions of different charge states originating both from surface contaminations and the sample material are found, and relative yields of several species were determined. Experimental evidence that points towards a predominantly electronic sputtering process is presented. For emitted Ti target atoms an additional nuclear sputtering component is suggested.

NleB/SseK effectors from Citrobacter rodentium, Escherichia coli, and Salmonella enterica display distinct differences in host substrate specificity

DEFF Research Database (Denmark)

El Qaidi, Samir; Chen, Kangming; Halim, Adnan

2017-01-01

proteins with N-acetyl-D-glucosamine to inhibit antibacterial and inflammatory host responses. NleB is conserved among the attaching/effacing pathogens enterohemorrhagic E. coli (EHEC), enteropathogenic E. coli (EPEC), and Citrobacter rodentium. Moreover, Salmonella enterica strains encode up to three Nle......B orthologs named SseK1, SseK2, and SseK3. However, there are conflicting reports regarding the activities and host protein targets among the NleB/SseK orthologs. Therefore, here we performed in vitro glycosylation assays and cell culture experiments to compare the activities and substrate specificities...... of these effectors. SseK1, SseK3, EHEC NleB1, EPEC NleB1, and C. rodentium NleB blocked TNF-mediated NF-κB pathway activation, whereas SseK2 and NleB2 did not. C. rodentium NleB, EHEC NleB1, and SseK1 glycosylated host glyceraldehyde 3-phosphate dehydrogenase (GAPDH). C. rodentium NleB, EHEC NleB1, EPEC NleB1...

Effects of Yizhi Capsule (益智胶囊) on Learning and Memory Disorder and β-amyloid Peptide Induced Neurotoxicity in Rats

Institute of Scientific and Technical Information of China (English)

WU Hang-yu; XU Jiang-ping; LI Lin; ZHU Bai-hua

2006-01-01

Objective: To explore the effects of Yizhi Capsule (益智胶囊, YZC) on learning and memory disorder and β-amyloid peptide induced neurotoxicity in rats. Methods: Various doses of YZC were administered to Sprague-Dawley (SD) rats for 8 consecutive days, twice a day. On the 8th day of the experiment,scopolamine hydrobromide was intraperitoneally injected to every rat and Morris water maze test and shuttle dark avoidance test were carried out respectively to explore the changes of learning and memory capacities in the rats. Besides, after the cerebral cortical neurons of newborn SD rats aged within 3 days were cultured in vitro for 7 days, drug serum containing YZC was added to the cultured neurons before or after β amyloid peptide25-35 (Aβ25-35) intoxication to observe the protective effect of YZC on neurotoxicity by MTT assay and to determine the LDH content in the supernatant. Results: Compared with those untreated with YZC, the rats having received YZC treatment got superiority in shorter time of platform seeking in Morris water maze test,as well as elongated latent period and less times of error in shuttle dark avoidance test. On the cultured neurons, YZC drug serum could effectively increase the survival rate of Aβ25-35 intoxicated neurons and reduce the LDH contents in cultured supernatant. Conclusion: YZC has an action of improving learning and memory disorder, and good protective effect on Aβ25-35 induced neurotoxicity in SD rats.

The domain-specific and temperature-dependent protein misfolding phenotype of variant medium-chain acyl-CoA dehydrogenase

NARCIS (Netherlands)

Jank, Johanna M.; Maier, Esther M.; Reiβ, Dunja D.; Haslbeck, Martin; Kemter, Kristina F.; Truger, Marietta S.; Sommerhoff, Christian P.; Ferdinandusse, Sacha; Wanders, Ronald J.; Gersting, Søren W.; Muntau, Ania C.

2014-01-01

The implementation of expanded newborn screening programs reduced mortality and morbidity in medium-chain acyl-CoA dehydrogenase deficiency (MCADD) caused by mutations in the ACADM gene. However, the disease is still potentially fatal. Missense induced MCADD is a protein misfolding disease with a

The cholinergic pathway alleviates acute oxygen and glucose deprivation induced renal tubular cell injury by reducing the secretion of inflammatory medium of macrophages

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Ming WU

2017-10-01

Full Text Available Objective To investigate the effects of cholinergic pathway on acute renal tubular cell injury induced by acute oxygen and glucose deprivation. Methods Rat kidney macrophages were isolated and cultured for constructing macrophages and renal epithelial cells co-cultivating model of oxygen-glucose deprivation (OGD, and the model cells were divided into three groups: OGD alone group, acetylcholine (ACh 100μmol/L+OGD group and ACh + galantamine (Gal 10μmol/L+OGD group. The cells underwent OGD treatment for 1 hour, and normally cultured for 24 hours. The expressions of TNF alpha, IL-1 beta, and IL-10 in supernatant fluid were detected by ELISA, the renal tubular cell viability was determined by MTT assay, the expression of acetylcholine esterase (AChE mRNA and protein were determined by RT-qPCR and Western blotting. The activity of AChE was determined by colorimetric method. Results The expressions of TNF alpha (pg/ml in OGD, Ach+OGD group, Ach+Gal+OGD groups were 140.2±44.81, 119.46±4.42 and 103.31±1.62 respectively (P0.05; The values of renal tubular cell proliferation were 55.02%±6.28%, 66.65%±6.47%, and 79.75%±4.22% respectively (P0.05; those of AchE protein were 0.66±0.07, 0.74±0.04 and 0.67±0.06 respectively (P>0.05; The activity of AChE (kU/L was 0.51±0.02, 0.35±0.05 and 0.32±0.04 respectively (P=0.001, 0.001 and 0.368. Conclusions ACh and Gal could inhibit the secretion of inflammatory mediators and cholinesterase activity and can reduce the acute hypoxic renal tubular cell injury. The modulation of the cholinergic pathway in macrophages may be the important treatment method for acute renal injury in the future. DOI: 10.11855/j.issn.0577-7402.2017.08.01

Exosome-mediated microRNA transfer plays a role in radiation-induced bystander effect.

Science.gov (United States)

Xu, Shuai; Wang, Jufang; Ding, Nan; Hu, Wentao; Zhang, Xurui; Wang, Bing; Hua, Junrui; Wei, Wenjun; Zhu, Qiyun

2015-01-01

Bystander effects can be induced through cellular communication between irradiated cells and non-irradiated cells. The signals that mediate this cellular communication, such as cytokines, reactive oxygen species, nitric oxide and even microRNAs, can be transferred between cells via gap junctions or extracellular medium. We have previously reported that miR-21, a well described DDR (DNA damage response) microRNA, is involved in radiation-induced bystander effects through a medium-mediated way. However, the mechanisms of the microRNA transfer have not been elucidated in details. In the present study, it was found that exosomes isolated from irradiated conditioned medium could induce bystander effects. Furthermore, we demonstrated plenty of evidences that miR-21, which is up-regulated as a result of mimic transfection or irradiation, can be transferred from donor or irradiated cells into extracellular medium and subsequently get access to the recipient or bystander cells through exosomes to induce bystander effects. Inhibiting the miR-21 expression in advance can offset the bystander effects to some extent. From all of these results, it can be concluded that the exosome-mediated microRNA transfer plays an important role in the radiation-induced bystander effects. These findings provide new insights into the functions of microRNAs and the cellular communication between the directly irradiated cells and the non-irradiated cells.

PENGIMBASAN KETAHANAN BIBIT PISANG AMBON KUNING TERHADAP PENYAKIT LAYU FUSARIUM DENGAN BEBERAPA JAMUR ANTAGONIS

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Loekas Soesanto dan Ruth Feti Rahayunia .

2011-11-01

Full Text Available Induced resistance of Ambon Kuning cultivar banana seedling to fusarium wilt with antagonistic fungi.  A research aiming at knowing the effect of antagonistic fungi supernatant on banana induced resistance, Fusarium wilt development, and banana growth was carried out from July up to December 2008. Randomized Block Design was used with four replicates. Treatments tested were control, with supernatant of Gliocladium virens, Trichoderma harzianum isolated from banana, ginger, and ginseng, Trichoderma koningii, and Fusarium equiseti, applied by injection to banana seedling corm and soaked for five minutes. Variables observed were incubation period, disease severity, Foc population density, germination inhibition, growth component, phenolic compound content, and supporting component. Result of the research indicated that the supernatant of G. virens, T. harzianum, and T. koningii could significantly induce resistance of the seedling showed by increasing the phenolic content such as glycoside, saponin, and tannin. The supernatant of all antagonistic fungi could effectively control the disease showed by lengthening incubation period as 48.71%, decreasing the disease severity as 53.57%, decreasing infection rate as 61.48%, increasing the antagonistic effectivity as 51.26%, decreasing the late population density as 45.35%, and decreasing the inoculum inside the plant as 60-100%. The extract could improve the seedling growth.

Chemical inducible promoter used to obtain transgenic plants with a silent marker

Science.gov (United States)

Aoyama, Takashi; Zuo, Jianru; Chua, Nam-Hai

2004-08-31

A chemically inducible promoter is described that may be used to transform plants, including tobacco and lettuce, with genes which are easily regulatable by adding the plants or plant cells to a medium containing an inducer of the promoter or by removing the plants or plant cells from such medium. The promoter described is one that is inducible by a glucocorticoid which is not endogenous to plants. Such promoters may be used with a variety of genes such as ipt or knotted1 to induce shoot formation in the presence of a glucocorticoid. The promoter may also be used with antibiotic or herbicide resistance genes which are then regulatable by the presence or absence of inducer rather than being constitutive. Other examples of genes which may be placed under the control of the inducible promoter are also presented.

Artemesia annua extract prevents glyoxal-induced cell injury in ...

African Journals Online (AJOL)

Purpose: To investigate the effect of Artemesia annua extract on glyoxal-induced injury in retinal microvascular endothelial cells (HRECs). Methods: HRECs were cultured in a medium containing 500 μM glyoxal or glyoxal plus 50μM Artemesia annua extract, or in the medium alone for 24 h. Apoptosis was analysed by flow ...

Photodynamic processes in LiCaAlF6:Ce3+ UV active medium

International Nuclear Information System (INIS)

Galiev, A I; Semashko, V V; Akhtyamov, O R; Shnaidman, S A; Marisov, M A; Shavelev, A A

2014-01-01

The objectives of this paper are experimental studies of pump-induced effects in LiCaAlF 6 : Ce 3+ single crystals and computer model elaboration and appropriated software package engineering. The elaborated experimental technique and software allow either to calculate the nonlinear absorption/gain characteristics of the active medium on the basis of known parameters, or to find its previously unknown parameters from pump-probe experimental dependences

Modelling antagonic effect of lactic acid eacteria supernatants on some pathogenic bacteria

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Augustus Caeser Franke Portella

2009-11-01

Full Text Available This work presents a statistical model of survival analysis for three pathogenic bacterial strains (Escherichia coli, Listeria monocytogenes and Staphylococcus aureus, when treated with neutralized and non-neutralized filtered supernatants broth from cultures of Lactobacillus acidhophilus, Lactobacillus rhamnosus and Lactobacillus sake. Survival analysis is a method employed to determine the period of time from an initial stage up to the occurrence of a particular event of interest, as death or a particular culture growth failure. In order to evaluate the potential efficacy of the ahead mentioned lactic acid bacteria when used as bioprotective starters in foods, experimental data were statistically treated and expressed by simple representative curves. Following the methodology of Cox and Kaplan-Meier, it was possible to make the selection of the best bioprotective lactic starter, as a predictive tool for evaluation of shelf life and prevention of eventual risks in fresh sausages and other similar food products.Este trabalho apresenta um modelo estatístico de análise de sobrevivência para três bactérias patogénicas (Escherichia coli, Listeria monocytogenes e Staphylococcus aureus, quando tratados com sobrenadantes filtrados neutralizado e não neutralizado provenientes de culturas de Lactobacillus acidhophilus, Lactobacillus rhamnosus e Lactobacillus sake. A Análise de sobrevivência é um método utilizado para determinar o período de tempo a partir de uma fase inicial até a ocorrência de um determinado evento de interesse, como a morte ou a inibição de uma particular cultura, a fim de avaliar a eficácia potencial das referidas bactérias lácticas quando usadas como bioproteção em alimentos. Os dados experimentais foram tratados estatisticamente, seguindo a metodologia de Cox e Kaplan-Meier e foi possível fazer a seleção dos melhores fermentos láticos bioprotectivos, como uma ferramenta para avaliação preditiva, vida de

ANKE, a new facility for medium energy hadron physics at COSY-Juelich

International Nuclear Information System (INIS)

Barsov, S.; Bechstedt, U.; Bothe, W.; Bongers, N.; Borchert, G.; Borgs, W.; Braeutigam, W.; Buescher, M.; Cassing, W.; Chernyshev, V.; Chiladze, B.; Dietrich, J.; Drochner, M.; Dymov, S.; Erven, W.; Esser, R.; Franzen, A.; Golubeva, Ye.; Gotta, D.; Grande, T.; Grzonka, D.; Hardt, A.; Hartmann, M.; Hejny, V.; Horn, L. van; Jarczyk, L.; Junghans, H.; Kacharava, A.; Kamys, B.; Khoukaz, A.; Kirchner, T.; Klehr, F.; Klein, W.; Koch, H.R.; Komarov, V.I.; Kondratyuk, L.; Koptev, V.; Kopyto, S.; Krause, R.; Kravtsov, P.; Kruglov, V.; Kulessa, P.; Kulikov, A.; Lang, N.; Langenhagen, N.; Lepges, A.; Ley, J.; Maier, R.; Martin, S.; Macharashvili, G.; Merzliakov, S.; Meyer, K.; Mikirtychiants, S.; Mueller, H.; Munhofen, P.; Mussgiller, A.; Nekipelov, M.; Nelyubin, V.; Nioradze, M.; Ohm, H.; Petrus, A.; Prasuhn, D.; Prietzschk, B.; Probst, H.J.; Pysz, K.; Rathmann, F.; Rimarzig, B.; Rudy, Z.; Santo, R.; Paetz Schieck, H.; Schleichert, R.; Schneider, A.; Schneider, Chr.; Schneider, H.; Schwarz, U.; Seyfarth, H.; Sibirtsev, A.; Sieling, U.; Sistemich, K.; Selikov, A.; Stechemesser, H.; Stein, H.J.; Strzalkowski, A.; Watzlawik, K.-H.; Wuestner, P.; Yashenko, S.; Zalikhanov, B.; Zhuravlev, N.; Zwoll, K.; Zychor, I.; Schult, O.W.B.; Stroeher, H.

2001-01-01

ANKE is a new experimental facility for the spectroscopy of products from proton-induced reactions on internal targets. It has recently been implemented in the accelerator ring of the cooler synchrotron COSY of the Forschungszentrum Juelich (FZ-Juelich), Germany. The device consists of three dipole magnets, various target installations and dedicated detection systems. It will enable a variety of hadron-physics experiments like meson production in elementary proton-nucleon processes and studies of medium modifications in proton-nucleus interactions

Stop Band Gap in Periodic Layers of Confined Atomic Vapor/Dielectric Medium

International Nuclear Information System (INIS)

Li Yuan-Yuan; Li Li; Lu Yi-Xin; Zhang Yan-Peng; Xu Ke-Wei

2013-01-01

A stop band gap is predicted in periodic layers of a confined atomic vapor/dielectric medium. Reflection and transmission profile of the layers over the band gap can be dramatically modified by the confined atoms and the number of layer periods. These gap and line features can be ascribed to the enhanced contribution of slow atoms induced by atom-wall collision, transient behavior of atom-light interaction and Fabry—Pérot effects in a thermal confined atomic system

27 CFR 19.914 - Medium plants.

Science.gov (United States)

2010-04-01

... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Medium plants. 19.914... OF THE TREASURY LIQUORS DISTILLED SPIRITS PLANTS Distilled Spirits For Fuel Use Permits § 19.914 Medium plants. Any person wishing to establish a medium plant shall make application for and obtain in...

Oral vaccines against diarrhea associated with enteropathogenic Escherichia coli strains based on genetically modified Bacillus subtilis strains

OpenAIRE

Wilson Barros Luiz

2010-01-01

O objetivo deste trabalho foi a construção de linhagens geneticamente modificadas de B. subtilis capazes de expressar porções de intimina, principal componente envolvido na capacidade de colonização de linhagens enteropatogênicas de Escherichia coli (EPEC), como estratégia vacinal de administração oral contra diarréias infecciosas. As vacinas desenvolvidas empregaram cinco regiões da intimina de EPEC e linhagens de B. subtilis capazes de expressar e acumular proteínas recombinantes no citopla...

The effects of L-arginine, D-arginine, L-name and methylene blue on channa striatus-induced peripheral antinociception in mice.

Science.gov (United States)

Zakaria, Zainul Amiruddin; Sulaiman, Mohd Rosian; Somchit, Muhammad Nazrul; Jais, Abdul Manan Mat; Ali, Daud Israf

2005-08-03

To determine the involvement of nitric oxide/cyclic guanosine monophosphate (NO/cGMP) pathway in aqueous supernatant of haruan (Channa striatus) fillet (ASH) antinociception using the acetic acid-induced abdominal constriction test. The ASH was prepared by soaking fresh haruan fillet in chloroform:methanol (CM) (2/1 (v/v)) for 72 h followed by evaporation of the upper layer supernatant to remove any solvent residues. The supernatant was then subjected to a freeze-drying process (48 h) followed by doses preparation. Subcutaneous (SC) administration of ASH alone (0.170, 0.426 and 1.704 mg/kg) exhibited a dose-dependent antinociception. On the other hand, 20 mg/kg (SC) of L-arginine and MB exhibited a significant nociception and antinociception, while D-arginine and L-NAME did not produce any effect at all. Pre-treatment with L-arginine was found to significantly reverse the three respective doses of ASH antinociception; pre-treatment with D-arginine did not produce any significant change in the ASH activity; pre-treatment with L-NAME only significantly increased the 0.170 and 0.426 mg/kg ASH antinociception; and pre-treatment with MB significantly enhanced the respective doses of ASH antinociception, respectively. Furthermore, co-treatment with L-NAME significantly enhanced the L-arginine reversal effect on 0.426 mg/kg ASH antinociception. In addition, MB significantly reversed the L-arginine nociception on 0.426 mg/kg ASH. These finding suggest ASH antinociception involves the nitric oxide (NO)/cyclic guanosine monophosphate (cGMP) pathway. The presence of NO was found to reverse ASH antinociceptive activity while blocking of cGMP system enhanced it.

The anomalous depolarization anisotropy in the central backscattering area for turbid medium with Mie scatterers

Science.gov (United States)

Wang, Xuezhen; Lai, Jiancheng; Song, Yang; Li, Zhenhua

2018-05-01

It is generally recognized that circularly polarized light is preferentially maintained over linearly polarized light in turbid medium with Mie scatterers. However, in this work, the anomalous depolarization anisotropy is reported in the backscattering area near the point of illumination. Both experimental and Monte Carlo simulations show preferential retention of linear polarization states compared to circular polarization states in a specific backscattering area. Further analysis indicates that the anomalous depolarization behavior in the specific area is induced by lateral scattering events, which own low circular polarization memory. In addition, it is also found that the size of the anomalous depolarization area is related to the transport mean free path of the turbid medium.

Cleavage of host cytokeratin-6 by lysine-specific gingipain induces gingival inflammation in periodontitis patients.

Directory of Open Access Journals (Sweden)

Salunya Tancharoen

Full Text Available Lysine-specific gingipain (Kgp is a virulence factor secreted from Porphyromonas gingivalis (P. gingivalis, a major etiological bacterium of periodontal disease. Keratin intermediate filaments maintain the structural integrity of gingival epithelial cells, but are targeted by Kgp to produce a novel cytokeratin 6 fragment (K6F. We investigated the release of K6F and its induction of cytokine secretion.K6F present in the gingival crevicular fluid of periodontal disease patients and in gingipain-treated rat gingival epithelial cell culture supernatants was measured by matrix-assisted laser desorption/ionization time-of-flight mass spectrometer-based rapid quantitative peptide analysis using BLOTCHIP. K6F in gingival tissues was immunostained, and cytokeratin 6 protein was analyzed by immunofluorescence staining and flow cytometry. Activation of MAPK in gingival epithelial cells was evaluated by immunoblotting. ELISA was used to measure K6F and the cytokines release induced by K6F. Human gingival fibroblast migration was assessed using a Matrigel invasion chamber assay.We identified K6F, corresponding to the C-terminus region of human cytokeratin 6 (amino acids 359-378, in the gingival crevicular fluid of periodontal disease patients and in the supernatant from gingival epithelial cells cultured with Kgp. K6F antigen was distributed from the basal to the spinous epithelial layers in gingivae from periodontal disease patients. Cytokeratin 6 on gingival epithelial cells was degraded by Kgp, but not by Arg-gingipain, P. gingivalis lipopolysaccharide or Actinobacillus actinomycetemcomitans lipopolysaccharide. K6F, but not a scrambled K6F peptide, induced human gingival fibroblast migration and secretion of interleukin (IL-6, IL-8 and monocyte chemoattractant protein-1. These effects of K6F were mediated by activation of p38 MAPK and Jun N-terminal kinase, but not p42/44 MAPK or p-Akt.Kgp degrades gingival epithelial cell cytokeratin 6 to K6F that, on

Neutron fluctuations in a medium randomly varying in time

International Nuclear Information System (INIS)

Lenard, Pal; Imre, Pazsit

2005-01-01

The master equation approach, which has traditionally been used for the calculation of neutron fluctuations in zero power systems with constant parameters, is extended to a case when the parameters of the system change randomly in time. We consider a forward type master equation for the probability distribution of the number of particles in a multiplying system whose properties jump randomly between two discrete states, both with and without an external source. The first two factorial moments are calculated, including the covariance. This model can be considered the unification of stochastic methods that were used either in a constant multiplying medium via the master equation technique, or in a fluctuating medium via the Langevin technique. In contrast to these methods, the one presented here can calculate the inherent noise in time-varying systems. The results obtained show a much richer characteristics of the zero power noise than that in constant systems. Even the concept of criticality has to be given a probabilistic interpretation. The asymptotic behaviour of the variance will be also qualitatively different from that in constant systems. The covariance of the neutron number in a subcritical system with a source, and the corresponding power spectrum, shows both the inherent and parametrically induced noise components. The results are relevant in medium power subcritical systems where the zero power noise is still significant, but they also have a bearing on all types of branching processes, such as evolution of biological systems, spreading of epidemics etc., which are set in a time-varying environment. (authors)

Neutron fluctuations in a medium randomly varying in time

Energy Technology Data Exchange (ETDEWEB)

Lenard, Pal [KFKI Atomic Energy Research Institute, Budapest (Hungary); Imre, Pazsit [Chalmers Univ. of Technology, Dept. of Nuclear Engineering, SE, Goteborg (Sweden)

2005-07-01

The master equation approach, which has traditionally been used for the calculation of neutron fluctuations in zero power systems with constant parameters, is extended to a case when the parameters of the system change randomly in time. We consider a forward type master equation for the probability distribution of the number of particles in a multiplying system whose properties jump randomly between two discrete states, both with and without an external source. The first two factorial moments are calculated, including the covariance. This model can be considered the unification of stochastic methods that were used either in a constant multiplying medium via the master equation technique, or in a fluctuating medium via the Langevin technique. In contrast to these methods, the one presented here can calculate the inherent noise in time-varying systems. The results obtained show a much richer characteristics of the zero power noise than that in constant systems. Even the concept of criticality has to be given a probabilistic interpretation. The asymptotic behaviour of the variance will be also qualitatively different from that in constant systems. The covariance of the neutron number in a subcritical system with a source, and the corresponding power spectrum, shows both the inherent and parametrically induced noise components. The results are relevant in medium power subcritical systems where the zero power noise is still significant, but they also have a bearing on all types of branching processes, such as evolution of biological systems, spreading of epidemics etc., which are set in a time-varying environment. (authors)

Effects of propofol on lipopolysaccharide-induced expression and release of HMGB1 in macrophages

Energy Technology Data Exchange (ETDEWEB)

Wang, T.; Wei, X.Y.; Liu, B.; Wang, L.J.; Jiang, L.H. [Department of Anesthesiology, the Third Affiliated Hospital, Zhengzhou University, Zhengzhou (China)

2015-02-24

This study aimed to determine the effects of different concentrations of propofol (2,6-diisopropylphenol) on lipopolysaccharide (LPS)-induced expression and release of high-mobility group box 1 protein (HMGB1) in mouse macrophages. Mouse macrophage cell line RAW264.7 cells were randomly divided into 5 treatment groups. Expression levels of HMGB1 mRNA were detected using RT-PCR, and cell culture supernatant HMGB1 protein levels were detected using enzyme-linked immunosorbent assay (ELISA). Translocation of HMGB1 from the nucleus to the cytoplasm in macrophages was observed by Western blotting and activity of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) in the nucleus was detected using ELISA. HMGB1 mRNA expression levels increased significantly in the cell culture supernatant and in cells after 24 h of stimulating RAW264.7 cells with LPS (500 ng/mL). However, HMGB1 mRNA expression levels in the P2 and P3 groups, which received 500 ng/mL LPS with 25 or 50 μmol/mL propofol, respectively, were significantly lower than those in the group receiving LPS stimulation (P<0.05). After stimulation by LPS, HMGB1 protein levels were reduced significantly in the nucleus but were increased in the cytoplasm (P<0.05). Simultaneously, the activity of NF-κB was enhanced significantly (P<0.05). After propofol intervention, HMGB1 translocation from the nucleus to the cytoplasm and NF-κB activity were inhibited significantly (each P<0.05). Thus, propofol can inhibit the LPS-induced expression and release of HMGB1 by inhibiting HMGB1 translocation and NF-κB activity in RAW264.7 cells, suggesting propofol may be protective in patients with sepsis.

Pre-equilibrium emission of nucleons from reactions induced by medium-energy heavy ions

International Nuclear Information System (INIS)

Korolija, M.; Holuh, E.; Cindro, N.; Hilscher, D.

1984-01-01

Recent data on fast-nucleon emission in heavy-ion-induced reactions are analysed successfully in terms of pre-equilibrium models; it is shown that the relevant parameters of those models preserve the physical meaning they have in light-ion-induced reactions. The initial exciton number obtained from a Griffin-plot analysis and the initial number of degrees of freedom, which is the relevant parameter of the modified HMB model, appear to be approximately equal for a given reaction at a given energy. It is inferred that, for heavy-ion reactions, the determination of such a parameter is substantially dominated by the centre-of-mass energy per nucleon above the Coulomb barrier, in contrast with the results of nucleon-induced reactions

Pions in the nuclear medium

International Nuclear Information System (INIS)

Chanfray, G.

1996-07-01

We discuss various aspects of pion physics in the nuclear medium. We first study s-wave pion-nucleus interaction in connection with chiral symmetry restoration and quark condensate in the nuclear medium. We then address the question of p-wave pion-nucleus interaction and collective pionic modes in nuclei and draw the consequences for in medium ππ correlations especially in the scalar-isoscalar channel. We finally discuss the modification of the rho meson mass spectrum at finite density and/or temperature in connection with relativistic heavy ion collisions

Anti-adherence potential of Enterococcus durans cells and its cell-free supernatant on plastic and stainless steel against foodborne pathogens.

Science.gov (United States)

Amel, Ait Meddour; Farida, Bendali; Djamila, Sadoun

2015-07-01

It is demonstrated that numerous bacteria are able to attach to surfaces of equipment used for food handling or processing. In this study, a strain of Enterococcus durans, originally isolated from a milking machine surface, was firstly studied for its biofilm formation potential on plastic and stainless steel supports. The strain was found to be a biofilm producer either at 25, 30 or 37 °C on polystyrene microtitre plates, with a best adherence level observed at 25 °C. En. durans showed a strong adhesion to stainless steel AISI-304. Antibacterial and anti-adherence activities of En. durans were tested against four foodborne pathogens (Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 25923, Pseudomonas aeruginosa ATCC 27853 and Listeria innocua CLIP 74915) which were shown as biofilm producers on both plastic and stainless steel. En. durans cells and cell-free culture supernatant showed a significant (P < 0.05) inhibition potential of the pathogens either on solid media or in broth co-cultures. Characterization of the antibacterial substances indicated their proteinaceous nature which assigned them most probably to bacteriocins group.

Optical recording medium

International Nuclear Information System (INIS)

Andriech, A.; Bivol, V.; Tridukh, G.; Tsiuleanu, D.

2002-01-01

The invention relates of the micro- and optoelectronics, computer engineering ,in particular, to tjhe optical information media and may be used in hilography. Summary of the invention consists in that the optical image recording medium, containing a dielectric substrates, onto one surface of which there are placed in series a transparent electricity conducting layer, a photo sensitive recording layer of chalcogenic glass and a thin film electrode of aluminium, is provided with an optically transparent protective layer, applied into the thin film electrode. The result of the invention consists in excluding the dependence of chemical processes course into the medium upon environmental conditions

[Differentiation of human umbilical cord derived mesenchymal stem cells into low immunogenic and functional hepatocyte-like cells in vitro].

Science.gov (United States)

Ren, Hong-ying; Zhao, Qin-jun; Xing, Wen; Yang, Shao-guang; Lu, Shi-hong; Ren, Qian; Zhang, Lei; Han, Zhong-chao

2010-04-01

To investigate the biological function of hepatocyte-like cells derived from mesenchymal stem cells that isolated from human umbilical cord UC-MSCs in vitro, and to detect the changes in the immunogenicity of the differentiated hepatocyte-like cells (DHC). Transdifferentiation of UC-MSCs into hepatic lineage in vitro was induced in modified two-step induction medium. The expressions of hepatic specific markers were detected by RT-PCR analysis and immunofluorescence staining at different time points after induction. The levels of albumin and urea in the supernatants of cultures were measured by enzyme-linked immunosorbent assay. Furthermore, the immunosuppressive property of DHC was detected by one-way mixed lymphocyte culture. The mRNA and proteins of alpha fetoprotein (AFP), albumin (ALB),and cytokeratin-19 (CK-19) were expressed in naive UC-MSCs at low levels. DHC highly expressed hepatic markers AFP, ALB, CK-19, and tryptophan 2, 3-dioxygenase 14 and 28 days after hepatic differentiation and were accompanied by an increased production of ALB and urea in supernatant in a time-dependent manner. DHC did not express human leukocyte antigen DR antigen and significantly decreased the lymphocyte proliferation. UC-MSCs are able to differentiate into functional hepatocyte-like cells in vitro, while the immunogenicity of DHC remains low.

A leukocyte activation test identifies food items which induce release of DNA by innate immune peripheral blood leucocytes.

Science.gov (United States)

Garcia-Martinez, Irma; Weiss, Theresa R; Yousaf, Muhammad N; Ali, Ather; Mehal, Wajahat Z

2018-01-01

Leukocyte activation (LA) testing identifies food items that induce a patient specific cellular response in the immune system, and has recently been shown in a randomized double blinded prospective study to reduce symptoms in patients with irritable bowel syndrome (IBS). We hypothesized that test reactivity to particular food items, and the systemic immune response initiated by these food items, is due to the release of cellular DNA from blood immune cells. We tested this by quantifying total DNA concentration in the cellular supernatant of immune cells exposed to positive and negative foods from 20 healthy volunteers. To establish if the DNA release by positive samples is a specific phenomenon, we quantified myeloperoxidase (MPO) in cellular supernatants. We further assessed if a particular immune cell population (neutrophils, eosinophils, and basophils) was activated by the positive food items by flow cytometry analysis. To identify the signaling pathways that are required for DNA release we tested if specific inhibitors of key signaling pathways could block DNA release. Foods with a positive LA test result gave a higher supernatant DNA content when compared to foods with a negative result. This was specific as MPO levels were not increased by foods with a positive LA test. Protein kinase C (PKC) inhibitors resulted in inhibition of positive food stimulated DNA release. Positive foods resulted in CD63 levels greater than negative foods in eosinophils in 76.5% of tests. LA test identifies food items that result in release of DNA and activation of peripheral blood innate immune cells in a PKC dependent manner, suggesting that this LA test identifies food items that result in release of inflammatory markers and activation of innate immune cells. This may be the basis for the improvement in symptoms in IBS patients who followed an LA test guided diet.

Identification of enteropathogenic Escherichia coli in children with acute diarrheic syndrome from Sucre State, Venezuela.

Science.gov (United States)

Michelli, Elvia; Millán, Adriana; Rodulfo, Hectorina; Michelli, Mirian; Luiggi, Jesús; Carreño, Numirin; De Donato, Marcos

2016-03-28

Diarrheagenic Escherichia coli is an important causative agent of acute diarrheic syndrome.  To identify clonal groups of enteropathogenic E. coli (EPEC), in 485 children with acute diarrhea aged 0 to 10 years attending health care centers in Arismendi, Benítez and Sucre municipalities, Sucre state, Venezuela, from March to December, 2011.  After obtaining the informed consent, stool samples were collected. Escherichia coli was identified using standard coproculture methods and serology with polyvalent and monovalent antisera. DNA was isolated, and eae (intimin) and bfpA (bundlin) genes were amplified through two multiplex polymerase chain reactions (PCR).  The presence of bacterial infection was determined in 39.6% of coprocultures. The prevalence of E. coli was 54.7%; 82.9% of these isolates were positive by serology for the evaluated serogroups and serotypes, which were mostly identified in children between 0 and 2 years (37.9%); 48.6% of E. coli strains amplified the eae gene; of these, 58.8% were classified as typical EPEC (eae+ y bfp+). EPEC II was the most common serogroup (38.7%), with predominance of typical EPEC (60%). In positive strains for eae gene, the β intimin allele was the most frequently identified (74.5%). Only four strains with O157:H7 serotype were identified, which showed no PCR amplification of the eae and bfpA genes.  This study showed the importance of molecular tests to identify diarrheagenic E. coli strains causing clinical conditions of varying severity.

Numerical simulations of mechanical and ignition-deflagration responses for PBXs under low-to-medium-level velocity impact loading.

Science.gov (United States)

Yang, Kun; Wu, Yanqing; Huang, Fenglei; Li, Ming

2017-09-05

An effective computational model is required to accurately predict the dynamic responses in accidental initiations of explosives. The present work uses a series of two-dimensional mechanical-chemical simulations performed via a hydrodynamic-code, DREXH-2D, to efficiently describe the mechanical and ignition-deflagration responses of cased cylindrical polymer-bonded explosives (PBXs) undergoing a low-to-medium-level impact (70-350m/s) in longitudinal direction. The ignition response was predicted based on an ignition criterion of effective plastic work. Slow burning and its growth to deflagration were described through a pressure-dependent reaction rate equation. The extreme value of effective plastic work was found to be useful to determine the ignition threshold velocity for PBXs. For low-level velocity impact, the incident stress wave reflection from lateral surfaces contributed to the formation of ignition regions. After the ignition, the deflagration was induced in the medium-level impact, and its violence was related to the shock strength. However, the low-strength stress wave only induced reaction at local regions, and sequent burning was no longer sensitive to the strength of incident wave. The predicted pressure and temperature results of PBXs were consistent with the medium-level impact tests performed by China Academy of Engineering Physics. Copyright © 2017 Elsevier B.V. All rights reserved.

Collaborative Manufacturing for Small-Medium Enterprises

Science.gov (United States)

Irianto, D.

2016-02-01

Manufacturing systems involve decisions concerning production processes, capacity, planning, and control. In a MTO manufacturing systems, strategic decisions concerning fulfilment of customer requirement, manufacturing cost, and due date of delivery are the most important. In order to accelerate the decision making process, research on decision making structure when receiving order and sequencing activities under limited capacity is required. An effective decision making process is typically required by small-medium components and tools maker as supporting industries to large industries. On one side, metal small-medium enterprises are expected to produce parts, components or tools (i.e. jigs, fixture, mold, and dies) with high precision, low cost, and exact delivery time. On the other side, a metal small- medium enterprise may have weak bargaining position due to aspects such as low production capacity, limited budget for material procurement, and limited high precision machine and equipment. Instead of receiving order exclusively, a small-medium enterprise can collaborate with other small-medium enterprise in order to fulfill requirements high quality, low manufacturing cost, and just in time delivery. Small-medium enterprises can share their best capabilities to form effective supporting industries. Independent body such as community service at university can take a role as a collaboration manager. The Laboratory of Production Systems at Bandung Institute of Technology has implemented shared manufacturing systems for small-medium enterprise collaboration.

Properties of the nuclear medium

International Nuclear Information System (INIS)

Baldo, M; Burgio, G F

2012-01-01

We review our knowledge on the properties of the nuclear medium that have been studied, over many years, on the basis of many-body theory, laboratory experiments and astrophysical observations. Throughout the presentation particular emphasis is placed on the possible relationship and links between the nuclear medium and the structure of nuclei, including the limitations of such an approach. First we consider the realm of phenomenological laboratory data and astrophysical observations and the hints they can give on the characteristics that the nuclear medium should possess. The analysis is based on phenomenological models, that however have a strong basis on physical intuition and an impressive success. More microscopic models are also considered, and it is shown that they are able to give invaluable information on the nuclear medium, in particular on its equation of state. The interplay between laboratory experiments and astrophysical observations is particularly stressed, and it is shown how their complementarity enormously enriches our insights into the structure of the nuclear medium. We then introduce the nucleon–nucleon interaction and the microscopic many-body theory of nuclear matter, with a critical discussion about the different approaches and their results. The Landau–Fermi liquid theory is introduced and briefly discussed, and it is shown how fruitful it can be in discussing the macroscopic and low-energy properties of the nuclear medium. As an illustrative example, we discuss neutron matter at very low density, and it is shown how it can be treated within the many-body theory. The general bulk properties of the nuclear medium are reviewed to indicate at which stage of our knowledge we stand, taking into account the most recent developments both in theory and experiments. A section is dedicated to the pairing problem. The connection with nuclear structure is then discussed, on the basis of the energy density functional method. The possibility of

Probing the intergalactic medium with fast radio bursts

International Nuclear Information System (INIS)

Zheng, Z.; Ofek, E. O.; Kulkarni, S. R.; Neill, J. D.; Juric, M.

2014-01-01

The recently discovered fast radio bursts (FRBs), presumably of extragalactic origin, have the potential to become a powerful probe of the intergalactic medium (IGM). We point out a few such potential applications. We provide expressions for the dispersion measure and rotation measure as a function of redshift, and we discuss the sensitivity of these measures to the He II reionization and the IGM magnetic field. Finally, we calculate the microlensing effect from an isolated, extragalactic stellar-mass compact object on the FRB spectrum. The time delays between the two lensing images will induce constructive and destructive interference, leaving a specific imprint on the spectra of FRBs. With a high all-sky rate, a large statistical sample of FRBs is expected to make these applications feasible.

The assessment of cytokines in Quantiferon supernatants for the diagnosis of latent TB infection in a tribal population of Melghat, India

Directory of Open Access Journals (Sweden)

Prachi R. Bapat

2015-07-01

Full Text Available Summary: The tuberculin skin test (TST and interferon-gamma release assays (IGRA, namely, the QuantiFERON-TB Gold test (QFT, remain the standard immunological diagnostic tools for latent tuberculosis (TB infection (LTBI. However, the sub-optimal detection rates of both of these tests are major impediments in recognizing the population at risk. This study was aimed at evaluating additional cytokines besides interferon-gamma (IFN-γ as biomarkers for improving LTBI diagnosis in the tribal population of Melghat, India. Seventy-four close TB contacts were stratified by QFT and TST results into: (i QFT+/TST+ (n = 26, (ii QFT+/TST− (n = 12, (iii QFT−/TST− (n = 35 and (iv QFT−/TST+ (n = 1 groups. A panel of cytokines (IL-6, IL-10, TNF-α and IL-2R was then evaluated in antigen-stimulated QFT cell-free culture supernatants using IMMULITE-1000, an automated immunoassay analyzer. Cytokine estimation showed significantly higher levels of IL-6 in the QFT+/TST+ group, while significantly higher levels of IL-10 were found in the QFT−/TST− group. Correlation analysis identified a positive correlation between IL-6 and the QFT response (r = 0.6723, P < 0.0001, while a negative correlation was seen between QFT and IL-10 expression (r = −0.3271, P = 0.0044. Similarly, IL-6 was positively correlated with TST levels (r = 0.6631, P < 0.0001, and conversely, a negative correlation was found between TST and IL-10 expression (r = −0.5698, P < 0.0001. The positive and negative predictive values of IL-6 were found to be 92.59 and 93.33%, respectively, and the positive and negative predictive values of IL-10 were 96.55 and 91.18%, respectively. No significant impact of the demographic characteristics on cytokine positivity was observed. Our preliminary results suggest that the evaluation of additional cytokines in QFT cell-free culture supernatants may be valuable for the identification of LTBI. Combining IL-6 and IL-10 with QFT and

Dynamics of the Content of Lactobacilli, Microbial Metabolites and Antimicrobial Activity of Growing Culture of Lactobacillus Plantarum 8P-A3

Directory of Open Access Journals (Sweden)

I. Yu. Chicherin

2013-01-01

Full Text Available The dynamics of the content of lactobacilli, microbial metabolites and antimicrobial activity of growing cultures of Lactobacillus plantarum 8Р-А3 was studied. Lactobacilli L. plantarum 8Р-А3 and test microorganisms isolated from the intestinal contents of patients with dysbacteriosis were used in experiments. Study of the component composition of growing culture supernatant of lactobacilli was carried out by gas liquid chromatography with mass selective detection. By 54 h of cultivation the content of viable microbial cells in the native culture of Lactobacillus achieves 3,0·109 in 1 mL without further increase during the cultivation. The principal component of lactobacilli culture medium possessing antibacterial activity is lactic acid. In addition to lactic acid, which accounts for 70% of the total metabolites, the culture medium and the supernatant contain salts of phosphoric acid (14% as well as amino acids, carboxylic acids, fatty acids, sugars and polyhydric alcohol constituting of up to 16% of the total metabolites. It is found that during the cultivation in liquid medium lactobacilli produce metabolites which possess antibacterial activity against pathogenic bacteria that cause intestinal infections.

Bacterial antagonists of fungal pathogens also control root-knot nematodes by induced systemic resistance of tomato plants.

Directory of Open Access Journals (Sweden)

Mohamed Adam

Full Text Available The potential of bacterial antagonists of fungal pathogens to control the root-knot nematode Meloidogyne incognita was investigated under greenhouse conditions. Treatment of tomato seeds with several strains significantly reduced the numbers of galls and egg masses compared with the untreated control. Best performed Bacillus subtilis isolates Sb4-23, Mc5-Re2, and Mc2-Re2, which were further studied for their mode of action with regard to direct effects by bacterial metabolites or repellents, and plant mediated effects. Drenching of soil with culture supernatants significantly reduced the number of egg masses produced by M. incognita on tomato by up to 62% compared to the control without culture supernatant. Repellence of juveniles by the antagonists was shown in a linked twin-pot set-up, where a majority of juveniles penetrated roots on the side without inoculated antagonists. All tested biocontrol strains induced systemic resistance against M. incognita in tomato, as revealed in a split-root system where the bacteria and the nematodes were inoculated at spatially separated roots of the same plant. This reduced the production of egg masses by up to 51%, while inoculation of bacteria and nematodes in the same pot had only a minor additive effect on suppression of M. incognita compared to induced systemic resistance alone. Therefore, the plant mediated effect was the major reason for antagonism rather than direct mechanisms. In conclusion, the bacteria known for their antagonistic potential against fungal pathogens also suppressed M. incognita. Such "multi-purpose" bacteria might provide new options for control strategies, especially with respect to nematode-fungus disease complexes that cause synergistic yield losses.

Bacterial antagonists of fungal pathogens also control root-knot nematodes by induced systemic resistance of tomato plants.

Science.gov (United States)

Adam, Mohamed; Heuer, Holger; Hallmann, Johannes

2014-01-01

The potential of bacterial antagonists of fungal pathogens to control the root-knot nematode Meloidogyne incognita was investigated under greenhouse conditions. Treatment of tomato seeds with several strains significantly reduced the numbers of galls and egg masses compared with the untreated control. Best performed Bacillus subtilis isolates Sb4-23, Mc5-Re2, and Mc2-Re2, which were further studied for their mode of action with regard to direct effects by bacterial metabolites or repellents, and plant mediated effects. Drenching of soil with culture supernatants significantly reduced the number of egg masses produced by M. incognita on tomato by up to 62% compared to the control without culture supernatant. Repellence of juveniles by the antagonists was shown in a linked twin-pot set-up, where a majority of juveniles penetrated roots on the side without inoculated antagonists. All tested biocontrol strains induced systemic resistance against M. incognita in tomato, as revealed in a split-root system where the bacteria and the nematodes were inoculated at spatially separated roots of the same plant. This reduced the production of egg masses by up to 51%, while inoculation of bacteria and nematodes in the same pot had only a minor additive effect on suppression of M. incognita compared to induced systemic resistance alone. Therefore, the plant mediated effect was the major reason for antagonism rather than direct mechanisms. In conclusion, the bacteria known for their antagonistic potential against fungal pathogens also suppressed M. incognita. Such "multi-purpose" bacteria might provide new options for control strategies, especially with respect to nematode-fungus disease complexes that cause synergistic yield losses.

BIFIDUM-MEDIUM FOR ISOLATION AND CULTIVATION OF BIFIDOBACTERIA

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L. V. Domotenko

2014-01-01

Full Text Available The comparative evaluation of Bifidum-medium quality (the nutrient medium for cultivation and selection bifidobacteria, dry and Blaurock medium has been performed. It was shown that Bifidum-medium supports the typical growth of the main types of bifidobacteria: Bifidobacterium bifidum, B. breve, B. adolescentis, B. infantis, B. longum. The comparable results for the efficacy and the activity accumulation of acid were obtained in the study of probiotic «bifidumbacterin » using both media. In studies of faecal material the medium concentration of bifidobacteria was found 8.15 lg CFU/g on Bifidum-medium and 6.68 lg CFU/g on Blaurock medium.