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1
Tamaru, Yutaka, Sadaharu Ui, Koichiro Murashima, Akihiko Kosugi, Helen Chan, Roy H. Doi, and Bo Liu. "Formation of Protoplasts from Cultured Tobacco Cells and Arabidopsis thaliana by the Action of Cellulosomes and Pectate Lyase from Clostridium cellulovorans." Applied and Environmental Microbiology 68, no. 5 (May 2002): 2614–18. http://dx.doi.org/10.1128/aem.68.5.2614-2618.2002.
Повний текст джерелаАнотація:
ABSTRACT The crude culture supernatants from Clostridium cellulovorans were tested for their ability to convert plant cells to protoplasts. The supernatants readily released protoplasts from cultured tobacco cells and Arabidopsis thaliana. The crude culture supernatant from pectin-grown cells was more active than supernatants from glucose-, cellobiose-, xylan-, and locust bean gum-grown cells. After removal of cellulosomes, the crude culture supernatant lost its protoplast formation activity. The protoplast formation activity of the crude culture supernatant from C. cellulovorans was more effective than those of commercial enzymes based on protein content.
2
Zavizion, Boris, A. John Bramley, and Ioannis Politis. "Effects ofStaphylococcus aureusproducts on growth and function of bovine mammary myoepithelial cellsin vitro." Journal of Dairy Research 62, no. 4 (November 1995): 577–86. http://dx.doi.org/10.1017/s0022029900031307.
Повний текст джерелаАнотація:
SUMMARYThe effects of culture supernatants conditioned by the growth ofStaphylococcus aureusM60 onin vitrogrowth and functional properties of bovine mammary myoepithelial cells were examined. Myoepithelial cell proliferation was reduced byStaph, aurexisM60 culture supernatants. Exposure of myoepithelial cells to culture supernatants of isogeneic mutants ofStaph, aureusM60 that produced either α or β toxins reduced proliferation, but to a lesser extent than supernatants from the wild type strain. Thus, α and β toxins may play some role in affecting myoepithelial cell proliferation. Of the cells tested, 42% contracted following addition of oxytocin (10–7M) in the culture medium. Treatment of myoepithelial cells for 15 min withStaph, aureusM60 supernatants, prior to addition of oxytocin in the culture medium, increased the number of cells that contracted to 92%. Exposure of cells for 3 h to the same supernatant, prior to addition of oxytocin in the culture medium, abolished oxytocin responsiveness, had no effect on immunolocalization of actin and vimentin, but affected the localization of α-actinin within myoepithelial cells. Treatment of myoepithelial cells for 3 h with a combination of purified staphylococcal proteinases XVI and XVII-B abolished oxytocin responsiveness and mimicked the effect of theStaph, aureusculture supernatant. We conclude thatStaph, aureusM60 culture supernatant affected proliferation and functional properties of myoepithelial cells.
3
Stachowiak, R., M. Lyzniak, B. K. Budziszewska, K. Roeske, J. Bielecki, G. Hoser, and J. Kawiak. "Cytotoxicity of Bacterial Metabolic Products, including Listeriolysin O, on Leukocyte Targets." Journal of Biomedicine and Biotechnology 2012 (2012): 1–9. http://dx.doi.org/10.1155/2012/954375.
Повний текст джерелаАнотація:
Bacterial toxins can exhibit anticancer activities. Here we investigated the anticancer effects of the listeriolysin O toxin produced byListeria monocytogenes. We found that supernatants ofListeria monocytogenesstrains (wild type, 1189, and 1190) were cytotoxic to the Jurkat cell line and human peripheral blood mononuclear cells (PBMC) in a concentration-dependent manner. The supernatant of strain 1044, not producing listeriolysin O, was inactive. The supernatants ofListeriastrains were also cytotoxic toward B cells of chronic leukemia patients, with no significant differences in activities between strains. We also tested supernatants ofBacillus subtilisstrains BR1-90, BR1-S, and BR1-89 producing listeriolysin O. BR1-S and BR1-89 were cytotoxic to PBMC and to Jurkat cells, the latter being more sensitive to the supernatants. BR1-90 was not hemolytic or cytotoxic to PBMC, but was cytotoxic to Jurkat cells in the concentration range of 10–30%, suggesting that listeriolysin O is selectively effective against T cells. Overall, the response of human peripheral blood mononuclear and human leukemia cell lines to bacteria supernatants containing listeriolysin O depended on the bacteria strain, target cell type, and supernatant concentration.
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Yu, F., Y. Itoyama, J. Kira, K. Fujihara, T. Kobayashi, T. Kitamoto, A. Suzumura, N. Yamamoto, Y. Nakajima, and I. Goto. "TNF-beta produced by human T lymphotropic virus type I-infected cells influences the proliferation of human endothelial cells and fibroblasts." Journal of Immunology 152, no. 12 (June 15, 1994): 5930–38. http://dx.doi.org/10.4049/jimmunol.152.12.5930.
Повний текст джерелаАнотація:
Abstract Human T lymphotropic virus type I (HTLV-I) is linked to adult T cell leukemia as well as to HTLV-I-associated myelopathy/tropical spastic paraparesis. In this report, we studied the effects of HTLV-I-infected cell supernatants on HUVEC, fibroblasts, and glioma cells. The HTLV-I-infected cell supernatants (HUT102 and MT-2) strongly inhibited the proliferation of HUVEC, although they enhanced the proliferation of the fibroblasts. Regarding the glioma cells, only the MT-2 supernatant showed weak inhibitory effects on the proliferation. However, the HTLV-I-uninfected cell supernatants showed no effects on these target cells. The biologic activities of both HUT102 and MT-2 supernatants were found to be dose dependent and were reduced by heat treatment at 100 degrees C for 5 min, but not at 56 degrees C for 30 min. These activities were not dependent on the concentrations of HTLV-I viral particles and were only minimally affected by the presence of anti-HTLV-I Abs. A bioassay of various cytokines revealed that the activity of TNF was much higher in the HUT102 and MT-2 supernatants than in the HTLV-I-uninfected cell supernatants (MOLT-4, Jurkat, and K-562). rTNF-alpha and rTNF-beta also showed strong inhibitory effects on HUVEC as well as on the enhancement of the fibroblast growth. With the use of Sephadex G-100 column chromatography, we obtained the highest activities from the 60- through 70-kDa fractions of the HUT102 supernatant and some activities from the 20- through 30-kDa fractions. The biologic activities of both the whole HUT102 supernatant and its active fractions were completely blocked by anti-TNF-beta mAb, although they were not blocked by anti-TNF-alpha mAb. In a Western blot assay, the 25- and 27-kDa bands of TNF-beta were shown clearly in the HUT102 supernatant, although no TNF-alpha bands appeared. These findings suggest that TNF-beta is present in either its oligomeric or monomeric form in the HTLV-I-infected cell supernatants and is also mainly responsible for the supernatants' effects on HUVECs and fibroblasts.
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Berger, M., EM Wetzler, and RS Wallis. "Tumor necrosis factor is the major monocyte product that increases complement receptor expression on mature human neutrophils." Blood 71, no. 1 (January 1, 1988): 151–58. http://dx.doi.org/10.1182/blood.v71.1.151.151.
Повний текст джерелаАнотація:
Abstract Interleukin-1 (IL-1) and other monocyte products have several important effects on the systemic response to infection in addition to their roles in lymphocyte stimulation. The present studies were carried out to determine whether products of stimulated monocytes activated circulating neutrophils (PMN) to increase expression of receptors for C3b (CR1) and C3bi (CR3), which are necessary for optimal margination, migration, and phagocytosis. Supernatants of human mononuclear cells that had been stimulated with lipopolysaccharide (LPS) or purified protein derivative (PPD) contained both tumor necrosis factor (TNF) and IL-1 and increased CR1 and CR3 expression on isolated PMNs. Supernatants of unstimulated cultures, media alone, or LPS or PPD alone had little or no effect. Supernatant effects were detectable at 1:3,000 final dilution and appeared to have a characteristic slow time course. These supernatants also caused dose- and time-dependent secretion of PMN granular constituents, but maximal receptor expression was accompanied by secretion of less than 10% of the cells' content of lysozyme and less than 16% of the B12 binding protein. Immunoadsorption studies showed that the supernatant's activity could be removed by anti- TNF but not by anti-IL-1. Recombinant IL-1 had no effect on receptor expression, but recombinant TNF increased CR1 and CR3 expression with kinetics similar to the supernatants. These results thus indicate that TNF is the major monocyte product that increases CR1 and CR3 expression on mature blood neutrophils. This would result in increased margination and phagocytic activity and may be an important systemic effect that would help the host eradicate infection.
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Berger, M., EM Wetzler, and RS Wallis. "Tumor necrosis factor is the major monocyte product that increases complement receptor expression on mature human neutrophils." Blood 71, no. 1 (January 1, 1988): 151–58. http://dx.doi.org/10.1182/blood.v71.1.151.bloodjournal711151.
Повний текст джерелаАнотація:
Interleukin-1 (IL-1) and other monocyte products have several important effects on the systemic response to infection in addition to their roles in lymphocyte stimulation. The present studies were carried out to determine whether products of stimulated monocytes activated circulating neutrophils (PMN) to increase expression of receptors for C3b (CR1) and C3bi (CR3), which are necessary for optimal margination, migration, and phagocytosis. Supernatants of human mononuclear cells that had been stimulated with lipopolysaccharide (LPS) or purified protein derivative (PPD) contained both tumor necrosis factor (TNF) and IL-1 and increased CR1 and CR3 expression on isolated PMNs. Supernatants of unstimulated cultures, media alone, or LPS or PPD alone had little or no effect. Supernatant effects were detectable at 1:3,000 final dilution and appeared to have a characteristic slow time course. These supernatants also caused dose- and time-dependent secretion of PMN granular constituents, but maximal receptor expression was accompanied by secretion of less than 10% of the cells' content of lysozyme and less than 16% of the B12 binding protein. Immunoadsorption studies showed that the supernatant's activity could be removed by anti- TNF but not by anti-IL-1. Recombinant IL-1 had no effect on receptor expression, but recombinant TNF increased CR1 and CR3 expression with kinetics similar to the supernatants. These results thus indicate that TNF is the major monocyte product that increases CR1 and CR3 expression on mature blood neutrophils. This would result in increased margination and phagocytic activity and may be an important systemic effect that would help the host eradicate infection.
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Jiménez-Vargas, N. N., M. Guzman Rodriguez, Y. Yu, E. Neary, A. E. Lomax, D. E. Reed, and S. Vanner. "A242 STOOL FROM IBS-D PATIENT WITH A HISTORY OF A DYSBIOTIC AND NON-DYSBIOTIC ONSET MODULATE NEURONAL EXCITABILITY VIA DIFFERENT MECHANISMS." Journal of the Canadian Association of Gastroenterology 5, Supplement_1 (February 21, 2022): 134–35. http://dx.doi.org/10.1093/jcag/gwab049.241.
Повний текст джерелаАнотація:
Abstract Background We have shown that Irritable Bowel Syndrome diarrhea-predominant (IBS-D) patients with a history of a dysbiotic-like onset have distinct stool metabolomic profiles versus those with a non-dysbiotic-like onset. IBS stool supernatants can sensitize mouse colonic afferent nerves via both histamine and proteases. However, it is unknown if stool supernatants from the two IBS-D subgroups modulate the excitability of nociceptive neurons via different neuroactive mediators Aims To evaluate whether there are differences in neuroactive mediators within stool supernatants from subgroups of IBS-D patients that can modulate the excitability of nociceptive neurons. Methods Stool samples from healthy control (HC) (N=5) and IBS-D patients with a dysbiotic (N=7) or non-dysbiotic-like (N=7) onset, was homogenized with Krebs solution and filtered. Proteolytic activity was assessed using casein as a substrate with and without protease inhibitors. Histamine was quantified by ELISA. DRG neurons from C57BL/6 mice were incubated overnight or acutely (30 min) with stool supernatants. Some neurons were pre-incubated with PAR2 (GB83, 10 μM) or H1R (pyrilamine, 1μM) antagonists prior to supernatant incubation. Changes in neuronal excitability were assessed with perforated patch-clamp by measuring the rheobase (current that elicits an action potential). Results Proteolytic activity in dysbiotic-like (57.9 U/μg, p<0.05) but not non-dysbiotic like (37.4 U/μg) stool supernatant was increased compared to HC (25.2 U/μg). Serine inhibitor decreased proteolytic activity of dysbiotic (46.6%, p<0.05) and non-dysbiotic (34.2%, p<0.01) supernatant whereas cysteine, aspartic and metalloproteases inhibitors had no effect. Histamine was increased 78% (p<0.05) in IBS-D compared to HC. No differences in proteolytic activity and histamine concentration between IBS-D subtypes were found. In patch-clamp recordings, overnight incubation with dysbiotic (19%, p<0.05) and non-dysbiotic (22%, p< 0.01) stool supernatant decreased rheobase of DRG neurons compared to HC. No difference in rheobase was observed between IBS-D subtypes. GB83 blocked the overnight actions of supernatants from both subgroups, but pyrilamine had no effect. In contrast, following acute incubation, pyrilamine blocked the hyperexcitability evoked by dysbiotic like supernatant (60pA vs 48pA, p<0.001) but had no effect on non-dysbiotic like supernatants. Conclusions Proteases in stool supernatants from IBS-D patients increase neuronal excitability but only those with a history of dysbiotic like onset acutely increase neuronal excitability through H1 receptors. These findings suggest that stool supernatants from subgroups of IBS-D may modulate nociceptor excitability via different mechanisms. Funding Agencies CIHRAmerican Neurogastroenterology and Motility Society
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Kanof, M. E., W. Strober, W. C. Kwan, N. A. O'Connell, and S. P. James. "CD4+ Leu-8+ T cell supernatant activity that inhibits Ig production." Journal of Immunology 147, no. 1 (July 1, 1991): 155–61. http://dx.doi.org/10.4049/jimmunol.147.1.155.
Повний текст джерелаАнотація:
Abstract The subpopulation of CD4+ T cells that expresses the Leu-8 peripheral lymph node homing receptor suppresses PWM-stimulated Ig synthesis. To determine the mechanism of this suppression, the immunoregulatory activity of culture supernatants obtained from peripheral blood CD4+ Leu-8+ T cells cultured with anti-CD3 mAb and PMA (Leu-8+ supernatant) was determined. Leu-8+ supernatant suppressed PWM-stimulated Ig synthesis in cultures containing non-T cells and CD4+ Leu-8- T cells. In contrast, the supernatant from CD4+ Leu-8- T cells did not suppress Ig synthesis. The inhibitory activity of CD4+ Leu-8+ T cell supernatants could not be accounted for by a deficiency or excess of IL-2, IL-4, IFN-gamma, IL-6, or PGE2. In studies examining the effect of CD4+ Leu-8+ supernatant on T cells, the supernatant did not alter either mitogen-induced proliferation or the helper function of CD4+ Leu-8- T cells. In studies examining the effect of CD4+ Leu-8+ supernatant on B cells, the supernatant inhibited Staphylococcus aureus Cowan I strain-induced B cell Ig secretion but not B cell proliferation. The suppressor activity of Leu-8+ supernatant was eliminated by protease treatment and was eluted by HPLC in two main peaks, with molecular sizes of 44 and 12 kDa. In summary, these studies indicate that supernatants from activated CD4+ Leu-8+ T cells directly suppress B cell Ig production.
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Wang, Yuehong, Tao Wang, Shanshan Xiao, Ruifang Mao, and Rui Lin. "Identify driver mutants in lung cancer by supernatants of pleural effusion and monitor resistance to targeted therapy." Journal of Clinical Oncology 37, no. 15_suppl (May 20, 2019): e14554-e14554. http://dx.doi.org/10.1200/jco.2019.37.15_suppl.e14554.
Повний текст джерелаАнотація:
e14554 Background: Malignant pleural effusion (MPE) from lung cancer is an attractive alternative for molecular testing due to advantages of non-invasiveness and less heterogeneity. Previously, cell pellet blocks of MPE are widely used. This study is to further address whether supernatants of MPE is a suitable source to identify key oncogenic mutants in lung cancer and provide more information for clinical molecular testing. Methods: MPE samples from 12 lung cancer patients were centrifuged to obtain supernatants and cell pellets, and DNA were extracted. The DNA samples were analyzed by a targeted next generation sequencing (NGS) panel using Illumina HiSeq platform. Results: In a pilot study, paired cancer tissue and MPE samples were obtained from 3 lung cancer patients and analyzed using a comprehensive 500-gene cancer panel. Nine mutants were identified both in the tissue samples and MPE samples. We then analyzed another 9 lung cancer patient samples to detect oncogenic mutants using an 18-gene panel. In total, 8 mutants including EGFR L858R, exon 19 deletion, or T790M mutants were identified in MPE samples. For the total 17 mutants from the 12 MPE samples, 10 mutants were observed in both matched supernatant and cell pellet of MPE, of which more pairs (6 out of 10) had supernatants with higher abundance of mutants than cell pellets. More importantly, 7 of the 17 mutants were only detected in the supernatants of the matched MPE. Taken together, these results suggest that supernatant of MPE is a more suitable source to detect key driver mutants of lung cancer. Interestingly, 2 patients had both tyrosine kinase inhibitor (TKI) resistant mutant T790M and TKI sensitive mutants detected in supernatants of MPE; both patients had prior treatment of EGFR TKI, consistent with the development of TKI resistant mutant and supporting the utility of supernatants of MPE in monitoring TKI resistance. Conclusions: This study suggest supernatant of MPE is a more suitable source for identifying key driver mutants for lung cancer and can also be used to monitor response to targeted therapy. The study provides evidence of using supernatant of MPE as alternative source for molecular testing and thus direct precise targeted therapy and effect surveillance.
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Neary, E., N. N. Jiménez-Vargas, A. E. Lomax, D. E. Reed, and S. Vanner. "A243 HISTAMINE H1-RECEPTOR ANTAGONISTS SUPPRESS THE HYPEREXCITABILITY OF NOCICEPTIVE NEURONS EXPOSED TO IBS-C PATIENT STOOL SUPERNATANTS." Journal of the Canadian Association of Gastroenterology 5, Supplement_1 (February 21, 2022): 135–36. http://dx.doi.org/10.1093/jcag/gwab049.242.
Повний текст джерелаАнотація:
Abstract Background Abdominal pain is the primary cause of morbidity in many chronic GI disorders such as IBS, but the molecular mechanisms contributing to pain signaling are unclear. Stool supernatants from patients with diarrhea-predominant IBS increase the excitability of DRG neurons compared to supernatants from healthy controls, suggesting that mediators in the stool may sensitize nociceptors. Additionally, histamine has been implicated as a mediator of hypersensitivity in IBS patients. However, it is unclear if stool supernatants from constipation-predominant IBS (IBS-C) patients affect the excitability of DRG neurons. Furthermore, it is unknown if specific neuro-mediators in stool such as histamine impact the excitability of nociceptive neurons in this subgroup of IBS patients. Aims To evaluate whether IBS-C stool supernatants induce nociceptive signaling in DRG neurons compared to healthy control (HC) stool supernatants. If so, to evaluate the role of histamine 1 (H1) receptors in DRG neuron nociceptive signaling initiated by mediators in IBS-C stool supernatants. Methods IBS-C (n=5) and HC (n=2) patient stool was collected, filtered, and dissolved with Krebs solution in a 1:8 (g/v) dilution. Dorsal root ganglion (DRG) neurons from C57BL/6 mice were incubated with HC or IBS-C stool supernatant for 30 minutes. To evaluate whether histamine in stool supernatants can sensitize H1 receptors on nociceptors, DRG neurons were pre-incubated with the H1-receptor antagonist pyrilamine (1μM, 30 min) before stool supernatants. Changes in DRG neuronal excitability were recorded using perforated patch-clamp techniques to measure the rheobase (minimum input current needed to elicit an action potential) and the resting membrane potential (RMP). Results In neurons incubated with IBS-C stool supernatants (n=28) the rheobase decreased (63%) compared to healthy controls (78 ± 13.7 pA; n=6). This effect was reversed in DRG neurons pre-incubated with the H1 receptor antagonist, pyrilamine, (n=26) (77 ± 5.9 pA, p<0.001) compared to neurons incubated with IBS-C supernatant alone (50 ± 5.13 pA; n=28). No changes were found in the RMP. The data were analyzed with the non-parametric Kruskal-Wallis test. Conclusions IBS-C stool supernatants increase the excitability of DRG neurons compared to HC. Furthermore, the H1-receptor antagonist pyrilamine inhibits the neuronal hyperexcitability evoked by mediators in IBS-C patient stool. These findings suggest that the neuroactive metabolite histamine may contribute to visceral pain experienced by patients with IBS-C. Further studies are needed to examine whether similar signaling to nociceptive neurons by stool supernatants occurs in other subtypes of IBS. Funding Agencies CAG, CIHR
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Estes, D. M., and J. M. Teale. "Biochemical and functional analysis of extracellular stress proteins of Mesocestoides corti." Journal of Immunology 147, no. 11 (December 1, 1991): 3926–34. http://dx.doi.org/10.4049/jimmunol.147.11.3926.
Повний текст джерелаАнотація:
Abstract Previous studies of the serum antibody response in mice to Mesocestoides corti infection indicated that molecules released by the parasite influenced the production of IgM and IgG1 to the exclusion of other isotypes. Two proteins isolated from M. corti culture supernatants were found to be homologous to the 70-kDa heat shock proteins (hsp70) and Escherichia coli GroEL families of stress proteins. The proliferative responses of splenic lymphocytes from infected mice were assessed to unfractionated M. corti supernatants as well as the 70- and 60-kDa stress protein homologs isolated from supernatants. Lymphocytes from infected mice respond to complete supernatant and both of the isolated p70 and p60 stress protein homologs. In addition, supernatant from M. corti cultures stimulates an in vitro antibody response restricted to IgM and IgG1; the same isotypes induced during infection. These results suggest that stress proteins play an integral part in the immune response to M. corti and the associated isotype restriction.
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Neary, E., N. N. Jiménez-Vargas, S. Osman, D. E. Reed, S. Vanner, and A. E. Lomax. "A234 EVALUATING THE EFFECT OF STOOL SUPERNATANTS FROM IBS PATIENTS AND HEALTHY CONTROLS ON THE EXCITABILITY OF DRG NEURONS." Journal of the Canadian Association of Gastroenterology 4, Supplement_1 (March 1, 2021): 283–84. http://dx.doi.org/10.1093/jcag/gwab002.232.
Повний текст джерелаАнотація:
Abstract Background Abdominal pain is commonly described in chronic disorders such as irritable bowel syndrome (IBS), but the underlying mechanisms are currently unclear. The stool metabolomic and microbiota profiles of IBS and healthy patients have shown distinct differences. Additionally, IBS stool supernatants have previously been demonstrated to induce hypersensitivity of nociceptive nerves in the ex vivo mouse colon, suggesting that mediators in the stool can sensitize nociceptors. However, the effects of healthy control (HC) or IBS patient stool supernatants on the excitability of DRG neurons have not been clarified. Aims To evaluate the effect of HC and IBS supernatant on DRG neurons. Methods HC (n=8 patients) or IBS (n=10 patients) stool was collected, dissolved and homogenized with bicarbonate-buffered Krebs solution at 37°C in a 1/10 dilution. DRG neurons from C57BL/6 mice were dissociated and incubated overnight with HC or IBS supernatant in a Krebs dissolution. Changes in DRG neuronal excitability were recorded using perforated patch-clamp techniques to measure the rheobase (amount of current needed to elicit an action potential). The effect of the IBS and HC stool supernatants on the resting membrane potential (RMP) was also recorded. Results Overnight incubations with supernatant of HC stool diluted in Krebs solution (n=28 neurons) did not significantly decrease the rheobase compared to control neurons (n=22) (62.7 ± 3.9 pA vs 64.2 ± 2.7 pA). In a parallel experiment, we evaluated the effect of IBS stool supernatants diluted in Krebs (n=52 neurons) and found that they significantly decreased the rheobase compared to the supernatant of HC diluted in Krebs and control neurons (52.3 ± 2.3; p<0.05). The data were analyzed with a one-way ANOVA and Tukey’s test. Incubations with IBS supernatant decreased the RMP compared to HC supernatant (-42.6 ± 0.6 mV vs. -46.0 ± 0.9 mV; p<0.01), which was calculated with an unpaired t-test. Conclusions These findings suggest that mediators in IBS stool increase the excitability of DRG neurons compared to HC stool supernatant, and thus may contribute to pain signaling in IBS patients. Funding Agencies CIHR
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Zampronio, A. R., M. C. C. Melo, C. A. A. Silva, I. R. Pelá, S. J. Hopkins, and G. E. P. Souza. "A pre-formed Pyrogenic Factor Released by Lipopolysaccharide Stimulated Macrophages." Mediators of Inflammation 3, no. 5 (1994): 365–73. http://dx.doi.org/10.1155/s0962935194000517.
Повний текст джерелаАнотація:
The aim of this study was to investigate the pyrogenic activity of factor(s) released by rat peritoneal macrophages following a brief stimulation with LPS. The effect of this factor on the number of circulating leukocytes and serum Fe, Cu and Zn levels, was also evaluated. The possibility that the content of interleukin (IL)-1β, IL-6 and tumour necrosis factor (TNF) in the supernatant could explain the observations was investigated. Supernatant produced over a period of 1 h by peritoneal macrophages, following a 30 min incubation with LPS at 37°C, was ultrafiltered through a 10 000 MW cut-off Amicon membrane, sterilized, and concentrated 2.5, 5, 10 and 20 times. The intravenous (i.v.) injection of this supernatant induced a concentration-dependent fever in rats with a maximal response at 2 h. The pyrogenic activity was produced by macrophages elicited with thioglycollate and by resident cells. The supernatants also induced neutrophilia and reduction in Fe and Zn 6 h after the injection. Absence of activity in boiled supernatants, or supernatants from macrophages incubated at 4°C with LPS, indicates that LPS was not responsible for the activity.In vitrotreatment with indomethacin (Indo), dexamethasone (Dex), or cycloheximide (Chx) did not modify the release of pyrogenic activity into the supernatant or its effects on the reduction in serum metal levels. Although Chx abolished the production of mediator(s) inducing neutrophilia, and Dex reduced the induction of IL-1β, TNF and IL-6, injection of the highest concentration of these cytokines detected in the supernatants did not induce fever.In vivotreatment with Dex, but not Indo, abolished the fever induced by the supernatant. These results suggest that macrophages contain pre-formed pyrogenic mediator(s), not related to IL-1β, IL-6 or TNF, that acts indirectly and independently of prostaglandtn. It also seems likely that the pyrogenic activity is related to the factor responsible for the reduction of serum Fe and Zn levels, but not the neutrophilia.
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Martins, Margarida, Mariana Henriques, Joana Azeredo, Sílvia M. Rocha, Manuel A. Coimbra, and Rosário Oliveira. "Morphogenesis Control in Candida albicans and Candida dubliniensis through Signaling Molecules Produced by Planktonic and Biofilm Cells." Eukaryotic Cell 6, no. 12 (November 2, 2007): 2429–36. http://dx.doi.org/10.1128/ec.00252-07.
Повний текст джерелаАнотація:
ABSTRACT Morphogenesis control by chemical signaling molecules is beginning to be highlighted in Candida biology. The present study focuses on morphogenic compounds produced in situ by Candida albicans and Candida dubliniensis during planktonic and biofilm growth that may at least partially substantiate the effect promoted by supernatants in morphogenesis. For both species, planktonic versus biofilm supernatants were analyzed by headspace-solid-phase microextraction and gas chromatography-mass spectrometry. Both planktonic cells and biofilm supernatants of C. albicans and C. dubliniensis contained isoamyl alcohol, 2-phenylethanol, 1-dodecanol, E-nerolidol, and E,E-farnesol. Alcohol secretion profiles were species, culture mode, and growth time specific. The addition of exogenous alcohols to the cultures of both species inhibited the morphological transition from the yeast to the filamentous form by up to 50%. The physiological role of these alcohols was put to evidence by comparing the effects of a 96-h cultured supernatant with synthetic mixtures containing isoamyl alcohol, 2-phenylethanol, E-nerolidol, and E,E-farnesol at concentrations determined herein. All synthetic mixtures elicited a morphological effect similar to that observed for the corresponding supernatants when used to treat C. albicans and C. dubliniensis cultures, except for the effect of the 96-h C. dubliniensis planktonic supernatant culture on C. albicans. Overall, these results reveal a group of alcohol extracellular signaling molecules that are biologically active with C. albicans and C. dubliniensis morphogenesis.
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Sayeed, Sameera, M. E. Fernandez-Miyakawa, Derek J. Fisher, Vicki Adams, Rachael Poon, Julian I. Rood, Francisco A. Uzal, and Bruce A. McClane. "Epsilon-Toxin Is Required for Most Clostridium perfringens Type D Vegetative Culture Supernatants To Cause Lethality in the Mouse Intravenous Injection Model." Infection and Immunity 73, no. 11 (November 2005): 7413–21. http://dx.doi.org/10.1128/iai.73.11.7413-7421.2005.
Повний текст джерелаАнотація:
ABSTRACT Clostridium perfringens type D enterotoxemias have significant economic impact by causing rapid death of several domestic animal species. Consequently, domestic animals are commonly vaccinated, at varying efficacy, with inactivated type D vegetative supernatants. Improved type D vaccines might become possible if the lethal toxins produced by type D isolates were characterized and the contributions of those toxins to supernatant-induced lethality were established. Therefore, the current study evaluated the presence of lethal toxins in supernatants prepared from late-log-phase vegetative cultures of a large collection of genotype D isolates. Under this growth condition, most genotype D isolates produced variable levels of at least three different lethal toxins, including epsilon-toxin (ETX). To model the rapid lethality of type D enterotoxemias, studies were conducted involving intravenous (i.v.) injection of genotype D vegetative supernatants into mice, which were then observed for neurotoxic distress. Those experiments demonstrated a correlation between ETX (but not alpha-toxin or perfringolysin O) levels in late-log-phase genotype D supernatants and lethality. Consistent with the known proteolytic activation requirement for ETX toxicity, trypsin pretreatment was required for, or substantially increased, the lethality of nearly all of the tested genotype D vegetative supernatants. Finally, the lethality of these trypsin-pretreated genotype D supernatants could be completely neutralized by an ETX-specific monoclonal antibody but not by an alpha-toxin-specific monoclonal antibody. Collectively, these results indicate that, under the experimental conditions used in the present study, ETX is necessary for the lethal properties of most genotype D vegetative supernatants in the mouse i.v. injection model.
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Sherris, D. I., and C. L. Sidman. "Distinction of B cell maturation factors from lymphokines affecting B cell growth and viability." Journal of Immunology 136, no. 3 (February 1, 1986): 994–98. http://dx.doi.org/10.4049/jimmunol.136.3.994.
Повний текст джерелаАнотація:
Abstract Supernatants from S26.5 helper T cells, autoimmune viable motheaten (mev/mev) mouse spleen cells, EL4 lymphoma cells, and recombinant DNA-derived interferon gamma (IFN-gamma), all of which display B cell maturation factor (BMF) activity, were assayed for effects on B cell proliferation alone and with Dextran Sulfate (DxS) and anti-immunoglobulin antibodies (alpha-Ig). Both EL4 and S26.5 supernatants showed BCGF-II (DxS co-stimulator) activity, whereas only EL4 supernatant had BCGF-I (alpha-Ig co-stimulator or BSF-I) activity. Supernatants from mev/mev spleen cells and recombinant DNA-derived IFN-gamma showed no activity in either assay. Fractionation of S26.5 supernatant by chromatofocusing showed a divergence of BMF activity (BMF-T, pIa of 6.0) from BCGF-II activity (pIa of 5.4), providing evidence for their physical nonidentity. IFN-gamma, which decreases B cell viability in culture, was separable from BMF-T by phenyl-Sepharose chromatography. BMF-T from S26.5 supernatant was separated from IFN-gamma and BCGF-II and was shown to induce B cell maturation without affecting B cell proliferation. The molecular characteristics of the purified BMF-T were pIa 6.0, Mr 55,000 by G-75 gel filtration, and Mr 16,000 by SDS-PAGE. These data demonstrate that several lymphokines (BMF) exist that mediate the maturation of B cells to active Ig secretion without stimulating B cell proliferation.
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Below, Sabine, Anne Konkel, Cathrin Zeeck, Christian Müller, Christian Kohler, Susanne Engelmann, and Jan-Peter Hildebrandt. "Virulence factors ofStaphylococcus aureusinduce Erk-MAP kinase activation and c-Fos expression in S9 and 16HBE14o- human airway epithelial cells." American Journal of Physiology-Lung Cellular and Molecular Physiology 296, no. 3 (March 2009): L470—L479. http://dx.doi.org/10.1152/ajplung.90498.2008.
Повний текст джерелаАнотація:
Part of the innate defense of bronchial epithelia against bacterial colonization is regulated secretion of salt, water, and mucus as well as defensins and cytokines involving MAP kinase activation and alterations in early gene expression. We tested two different types of immortalized human airway epithelial cells (S9, 16HBE14o-) for activation of Erk-type MAP kinases and for expression of c-Fos on treatment with Staphylococcus aureus culture supernatants from the stationary growth phase [optical density (OD)540nm= 10] or with recombinant S. aureus hemolysins A and B (Hla, Hlb). OD10 supernatants activated Erk-type MAP kinases and c-Fos expression in a concentration-dependent manner. Hla induced Erk-type kinase phosphorylation in S9 but not in 16HBE14o- cells. Hlb induced Erk activation in either cell type. Basal and stimulated levels of Erk-type MAP kinase phosphorylation were sensitive to the Mek1 inhibitor PD-98059, indicating that the bacterial products activated the entire signaling cascade that coregulates IL-8 induction and secretion. While c-Fos expression was enhanced by OD10 supernatants, Hla, and Hlb in S9 cells, 16HBE14o- cells responded to OD10 supernatant and Hlb but not to Hla. In S9 cells, PD-98059 suppressed c-Fos upregulation by OD10 supernatant, Hla, or Hlb, indicating that c-Fos expression requires activation of Erk-type MAP kinases. In 16HBE14o- cells, however, c-Fos expression by OD10 supernatant was sensitive to PD-98059, while that induced by Hlb was not. This indicates that ingredients of OD10 supernatants other than Hla or Hlb are activating Erk-type MAP kinases in 16HBE14o- cells and that other intracellular signaling systems apart from Erk-type MAP kinases contribute to Hlb-mediated regulation of c-Fos. Thus interaction of bacterial factors with airway epithelial cells may be highly cell type specific.
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Gaskin, F., B. S. Kingsley, and S. M. Fu. "Autoantibodies to neurofibrillary tangles and brain tissue in Alzheimer's disease. Establishment of Epstein-Barr virus-transformed antibody-producing cell lines." Journal of Experimental Medicine 165, no. 1 (January 1, 1987): 245–50. http://dx.doi.org/10.1084/jem.165.1.245.
Повний текст джерелаАнотація:
Multiple EBV-transformed B cell lines were established from five patients with a clinical diagnosis of Alzheimer's disease (AD) and six age-matched controls. The supernatants were screened for antibody activity against SDS-treated isolated neurofibrillary tangles (NFT). Reactive supernatants were identified from both the AD and control group. The frequencies of anti-NFT antibody-secreting lines were 6.3 and 1.6% for the AD and the control groups, respectively. A proportion of these supernatants also stained NFT in situ and neurons and/or glia in sections of the frontal and the temporal cortexes of autopsied AD and normal brains, as well as cells from three cell lines (HeLa, fibroblast, and neuroblastoma). Several patterns of staining were revealed by these supernatants, indicating different reactive antigens. One supernatant stained NFT and astrocytes in sections from AD brains. It did not stain sections from two normal brains. This cell line is the result of the immortalization of a circulating B cell making antibody specific for an antigen in AD. The present approach may provide new insights in the pathogenesis of AD.
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Akimoto, Tetsu, and Marc R. Hammerman. "Microvessel formation from mouse aorta is stimulated in vitro by secreted VEGF and extracts from metanephroi." American Journal of Physiology-Cell Physiology 284, no. 6 (June 1, 2003): C1625—C1632. http://dx.doi.org/10.1152/ajpcell.00436.2002.
Повний текст джерелаАнотація:
We have demonstrated that during culture under 5% O2, the addition of recombinant human VEGF or FGF2 to mouse embryonic aorta explants (thoracic level to lateral vessels supplying the mesonephros and metanephros) stimulates microvessel formation. Here we show that microvessel formation is also stimulated by addition to explants of supernatants obtained from metanephroi grown in serum-free organ culture or of metanephroi extracts. Supernatants and extracts from metanephroi grown under hypoxic conditions are more stimulatory than supernatants/extracts from metanephroi grown in room air. VEGF and FGF2 can be detected by using immunohistochemistry in developing nephrons in the cultured renal anlagen. Metanephroi supernatants contain more VEGF if renal anlagen are grown under hypoxic conditions than if they are grown in room air. Metanephros supernatant-stimulated microvessel formation is completely inhibited by soluble sFlt-1 fusion protein or anti-VEGF antibodies (αVEGF). Extract-stimulated microvessel formation is inhibited by αVEGF or anti-FGF2 antibodies, or both. We conclude that metanephroi produce growth factors including VEGF and FGF that enhance microvessel formation from embryonic thoracic aorta in vitro.
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Kubelka, C. F., A. Ruppel, P. H. Krammer, and D. Gemsa. "Killing of schistosomula of Schistosoma mansoni by macrophages: induction by T-cell clone-derived lymphokines and interferon-gamma." Parasitology 92, no. 2 (April 1986): 325–36. http://dx.doi.org/10.1017/s003118200006409x.
Повний текст джерелаАнотація:
SUMMARYThe induction of schistosomulicidal activity of peritoneal macrophages by concanavalin A-stimulated supernatants from long-term T-cell clones and by interferon-gamma (IFN-γ) was investigated in detail. Optimal conditions of in vitro macrophage activation by T-cell clone supernatants were established. Macrophages from 13-week S. mansoni-infected mice responded to lymphokine activation as well as resident mnacrophages from uninfecteci mice. IFN-γ was shown to play an essential role in induction of schistosomulicidal macrophage activity: recombinant IFN-γ at high concentration could induce schistosomula killing, and an anti-IFN-γ antiserum inhibited the induction ofschistosomulicidal activity by T-cell clone supernatants. Our data also indicate that macrophage activation could be obtained by IFN-γ in synergy with other lymphokines in the supernatant of long-term T-cell clones. Macrophages from mice injected with T-cell clone supernatants were primed in vivo and triggered to kill schistosomula in vitro in the presence of lipopolysaccharide (LPS). The data demonstrate that lymphokines produced by T-cell clones and, in particular, IFN-γ can participate in the activation of schistosomulicidal macrophages.
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Subbiah, Murugan, Shannon M. Mitchell, Jeffrey L. Ullman та Douglas R. Call. "β-Lactams and Florfenicol Antibiotics Remain Bioactive in Soils while Ciprofloxacin, Neomycin, and Tetracycline Are Neutralized". Applied and Environmental Microbiology 77, № 20 (19 серпня 2011): 7255–60. http://dx.doi.org/10.1128/aem.05352-11.
Повний текст джерелаАнотація:
ABSTRACTIt is generally assumed that antibiotic residues in soils select for antibiotic-resistant bacteria. This assumption was tested by separately adding 10 different antibiotics (≥200 ppm) to three soil-water slurries (silt-loam, sand-loam, and sand; 20% soil [wt/vol]) and incubating mixtures for 24 h at room temperature. The antibiotic activity of the resultant supernatant was assessed by culturing a sensitiveEscherichia colistrain in the filter-sterilized supernatant augmented with Luria-Bertani broth. We found striking differences in the abilities of supernatants to suppress growth of the indicatorE. coli. Ampicillin, cephalothin, cefoxitin, ceftiofur, and florfenicol supernatants completely inhibited growth while bacterial growth was uninhibited in the presence of neomycin, tetracycline, and ciprofloxacin supernatants. High-performance liquid chromatography (HPLC) analysis demonstrated that cefoxitin and florfenicol were almost completely retained in the supernatants, whereas tetracycline and ciprofloxacin were mostly removed. Antibiotic dissipation in soil, presumably dominated by adsorption mechanisms, was sufficient to neutralize 200 ppm of tetracycline; this concentration is considerably higher than reported contamination levels. Soil pellets from the tetracycline slurries were resuspended in a minimal volume of medium to maximize the interaction between bacteria and soil particles, but sensitive bacteria were still unaffected by tetracycline (P= 0.6). Thus, residual antibiotics in soil do not necessarily exert a selective pressure, and the degree to which the pharmaceutical remains bioactive depends on the antibiotic. Efforts to control antibiotic contamination would be better directed toward compounds that retain biological activity in soils (e.g., cephalosporins and florfenicol) because these are the antibiotics that could exert a selective pressure in the environment.
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Motta, Juliana, Aline Sperandio, Morgana Castelo-Branco, Massimo Locati, and Vivian Rumjanek. "Evaluation of myeloid cell suppression related to cancer cell released products (TUM4P.913)." Journal of Immunology 192, no. 1_Supplement (May 1, 2014): 138.14. http://dx.doi.org/10.4049/jimmunol.192.supp.138.14.
Повний текст джерелаАнотація:
Abstract Dendritic cells (DCs) and macrophages may recognize tumor antigens and initiate an antitumoral response before and after the formation of tumor mass. However, the secretion of cytokines and factors that inhibit myeloid cells is a mechanism used by tumor cells to escape from immune attack. Our aim was to understand how tumor cells modulate the generation of murine macrophages and human DCs. For this, we collected bone marrow cells from C57Bl/6 mice and differentiated with M-CSF in the presence of 3LL (Lewis lung carcinoma) or MN/MCA (fibrosarcoma) supernatants. Some cells were activated with IFN-γ and LPS to obtain M1 macrophages or IL-4 to M2. Data showed increased expression of arginase-1 mRNA in macrophages differentiated with MN/MCA supernatant and it happened both in M1 and M2. About 20% of cells proliferate in the presence of supernatants, even in the absence of M-CSF and they express higher level of CD16. Moreover, we differentiated human DCs from monocytes with IL-4 and GM-CSF in the presence of K562 (chronic myeloid leukemia) supernatants. TNF-α was added to activate the cells. It was observed higher percentage of cells expressing CD80 when the differentiation was done with K562 supernatant. Higher secretion of IL-10 and lower production of IL-12 were found in cells differentiated with K562 supernatant and later activated. Therefore, the development and function of macrophages and DCs are affected by tumor products and both cells present some suppressive features.
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Freedman, D. O., S. Parker-Cook, M. C. Maia e Silva, C. Braga, and A. Maciel. "Very late antigen-4/vascular cell adhesion molecule-1 (VLA-4/VCAM-1) pathway is involved in the transendothelial migration of lymphocytes in bancroftian filariasis." Journal of Immunology 156, no. 8 (April 15, 1996): 2901–8. http://dx.doi.org/10.4049/jimmunol.156.8.2901.
Повний текст джерелаАнотація:
Abstract The role of endothelial vascular cell adhesion molecule-1 (VCAM-1) in the trafficking of lymphocytes from the vascular circulation through endothelium and into sites of filarial inflammation was investigated in asymptomatic microfilaremic (MF; n = 16) and in patients with filaria-associated lymphatic pathology (LP; n = 23). When compared, by human umbilical vein endothelial cell ELISA, with PBMC supernatants generated from MF patients, filarial Ag (Brugia malayi adult Ag (BmA))-stimulated supernatants from LP patients stimulated over 50% more VCAM-1 (lymphatic pathology (LP) = 0.377 relative ELISA units vs MF = 0.246; p = 0.02). No difference was seen between patient groups with nonfilarial Ag (streptolysin O)-stimulated supernatant (LP = 0.160 relative ELISA units vs MF = 0.166). BmA-stimulated supernatants from LP patients stimulated significantly more T cell migration (percentage of relative migration LP = 79.9% vs MF = 25.8%; p = 0.01) through tightly confluent human umbilical vein endothelial cell monolayers cultured on collagen gels. Anti-VCAM-1 inhibited the increased T cell migration induced by BmA-stimulated supernatants by 97.7%, anti-VLA-4 by 74.7%, and blockade of NF-kappa B-dependent VCAM-1 transcription with 50 microM pyrrolidine dithiocarbamate resulted in 87.7% inhibition. Biopsies from 87.5% of the LP patients, but only 38.5% of the MF patients, demonstrated VCAM-1 on vascular endothelium. BmA-stimulated supernatants pretreated with anti-IL-1 alone resulted in VCAM-1 expression that was 23.7% of that with untreated supernatants while anti-IL-4, anti-IFN-gamma, or anti-TNF alone had essentially no effect on VCAM-1 expression.
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Cannon, Jennifer S., Dolores Ciufo, Anita L. Hawkins, Constance A. Griffin, Michael J. Borowitz, Gary S. Hayward, and Richard F. Ambinder. "A New Primary Effusion Lymphoma-Derived Cell Line Yields a Highly Infectious Kaposi's Sarcoma Herpesvirus-Containing Supernatant." Journal of Virology 74, no. 21 (November 1, 2000): 10187–93. http://dx.doi.org/10.1128/jvi.74.21.10187-10193.2000.
Повний текст джерелаАнотація:
ABSTRACT A primary effusion lymphoma (PEL) cell line, JSC-1, that yields highly infectious Kaposi's sarcoma herpesvirus (KSHV) supernatants was established from the ascitic fluid of a human immunodeficiency virus-positive patient. Flow cytometry showed strong expression of CD45 and lambda light-chain restriction. Southern blot hybridization showed immunoglobulin heavy-chain gene rearrangements in the tumor and the resultant cell line consistent with B-cell lineage. Expression of viral genes was assessed by reverse transcription-PCR and immunohistochemistry. Only latent Epstein-Barr virus (EBV) gene expression was detected, and this was at a low level. In contrast, lytic and latent KSHV gene expression were detected. Tetradecanoyl phorbol acetate and butyrate upregulated KSHV lytic expression, but not EBV lytic expression. Viral supernatant from JSC-1 was much more efficient at infecting primary human dermal microvascular endothelial cells (DMVECs) with KSHV than supernatants from BC-3 or BCP-1 PEL cell lines. Quantitation of viral yields produced by the PEL lines showed at least 2 orders of magnitude more DNase I-resistant KSHV DNA in the JSC-1 supernatant compared to BC-3 or BCP-1 supernatants. KSHV infection in DMVECs was associated with a change from a cobblestone to a spindle shape, LANA expression, and an increased number of mitoses.
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Rubinstein, Israel, Xiao-Pei Gao, Sergei Pakhlevaniants, and Dolphine Oda. "Smokeless tobacco-exposed oral keratinocytes increase macromolecular efflux from the in situ oral mucosa." American Journal of Physiology-Regulatory, Integrative and Comparative Physiology 274, no. 1 (January 1, 1998): R104—R111. http://dx.doi.org/10.1152/ajpregu.1998.274.1.r104.
Повний текст джерелаАнотація:
The purpose of this study was to determine whether supernatants of cultured human oral keratinocytes (HOK) exposed to an aqueous extract of smokeless tobacco (STE) increase macromolecular efflux from the oral mucosa in vivo and, if so, whether bradykinin mediates in part this response. Subconfluent monolayers of HOK were incubated with STE or media, and supernatants were collected 24, 48, and 72 h thereafter. Using intravital microscopy, we found that suffusion of supernatants of STE- but not media-exposed HOK elicited significant concentration- and time-dependent increases in efflux of fluorescein isothiocyanate-labeled dextran (mol mass 70 kDa) from the in situ hamster cheek pouch ( P < 0.05). These effects were significantly attenuated by HOE-140 and NPC-17647 but not by des-Arg9,[Leu8]-bradykinin. Proteolytic activity was increased in supernatants of STE- but not media-exposed HOK. However, a mixture of leupeptin, Bestatin, anddl-2-mercaptomethyl-3-guanidinoethylthiopropanoic acid had no significant effects on HOK supernatant-induced responses. Collectively, these data suggest that oral keratinocytes modulate smokeless tobacco-induced increase in macromolecular efflux from the in situ oral mucosa in part by elaborating proteases that may account for local bradykinin production.
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Coffman, R. L., and J. Carty. "A T cell activity that enhances polyclonal IgE production and its inhibition by interferon-gamma." Journal of Immunology 136, no. 3 (February 1, 1986): 949–54. http://dx.doi.org/10.4049/jimmunol.136.3.949.
Повний текст джерелаАнотація:
Abstract The addition of concanavalin A-stimulated supernatants of the helper T cell clone, D9.1, to cultures of lipopolysaccharide (LPS)-stimulated T-depleted mouse spleen cells caused more than a 100-fold increase in immunoglobulin (Ig) E production. These supernatants cause a 10-fold to 15-fold increase in IgG1, a fivefold to 10-fold increase in IgA, and a fivefold to 10-fold decrease in IgG3. These effects are optimal when the supernatants are added 1 to 2 days after stimulation with LPS. Cells from mouse strains that normally give little or no IgE response in vivo give normal IgE levels in response to LPS plus the supernatant of Concanavalin A-stimulated D9.1 cells in vitro. The enhancement of both IgE and IgG1 can be completely inhibited by relatively low concentrations of interferon-gamma (IFN-gamma). Both the IgE-enhancing activity and IFN-gamma act directly upon purified B cells.
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Barreto, Marliton R., Leandro L. Loguercio, Fernando H. Valicente, and Edilson Paiva. "Insecticidal activity of culture supernatants from Bacillus thuringiensis Berliner strains against Spodoptera frugiperda Smith (Lepidoptera: Noctuidae) larvae." Anais da Sociedade Entomológica do Brasil 28, no. 4 (December 1999): 675–85. http://dx.doi.org/10.1590/s0301-80591999000400010.
Повний текст джерелаАнотація:
Novel vegetative insecticidal proteins (Vips) identified in the supernatant of Bacillus thuringiensis (B.t.) cultures have shown to provide adequate control over a wide spectrum of economically important crop pests. To evaluate the potential applicability of these proteins against fall armyworm (Spodoptera frugiperda Smith) larvae, the most important insect pest for tropical maize, the characteristics and mortality effects of culture supernatants from five B.t. strains were investigated. Striking differences among strains were detected, not only in terms of efficiency in killing the insect, but also regarding to mortality effects of heated and non-heated supernatants, which were used to distinguish the heat-sensitive protein-derived insecticidal fraction from a thermostable one, with a non-protein nature (b-exotoxinas). The qualitative, quantitative and temporal patterns of total protein secretion in the medium (supernatant) were assessed through spectrophotometry and polyacrylamide gel electrophoresis. The strains showed remarkably distinct rates of growth and timing for protein secretion relative to cell density in culture. Moreover, the electrophoretic-banding patterns also varied in a strain-specific manner, both in denaturing and non denaturing conditions. Polypeptides displaying a molecular weight that is very close to the expected for previously identified Vip3A proteins were found for the strains with high supernatant-mortality ratios. The data suggest the feasibility and usefulness of searching for protein-derived (Vip-like) insecticidal fractions in B.t. supernatants as a mean of developing especific and efficient alternatives of biological control to be employed in integrated pest management programs of S. frugiperda in tropical maize.
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Yuan, Fahu, Yufei Liu, Qian Gui, Qiuyi Huang, Qianyu Li, Xuping Yang, Lixin Qiu, Jinmei Feng, and Xiji Shu. "Efficacy of Bifidobacterium animalis subsp. lactis BB-12 against Giardia duodenalis trophozoites: an experimental study." E3S Web of Conferences 233 (2021): 02048. http://dx.doi.org/10.1051/e3sconf/202123302048.
Повний текст джерелаАнотація:
Giardia duodenalis, formerly known as Giardia lamblia, is an important zoonotic protozoan parasite. It mainly infects the intestines of humans, dogs, cats and domestic animals, causing diarrhea, abdominal pain, indigestion and weight loss. At present, all the clinical drugs for the treatment of Giardia have problems such as side effects and drug resistance to varying degrees, and the development of new drugs for the treatment of Giardia is still a hot issue. There is growing interest in using probiotics as an anti-intestinal parasite strategy. The present study aimed to assess the effect of supernatants of Bifidobacterium Animalis Subsp. lactis BB-12 on giardia the growth of giardia trophozoites. In this study, the Bifidobacterium animalis subsp. lactis BB-12 were cultured in BBL liquid medium, and the effects of the supernatants on the growth and adhesion of trophozoites of Giardia were observed. The results showed that the growth of Giardia flagellate was significantly inhibited by the supernatant. The influence of the supernatant on the morphology of the trophozoites was observed by microscope, and it was found that the surface of the trophozoites was uneven, the shape was atrophied, the surface cell membrane was broken to some extent, and the contents were spilt. In summary, the results of this study suggest that the fresh-cultured supernatants of the probiotic Bifidobacterium Animalis subsp. lactis BB-12 have anti-Giardia effects.
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Gamba, Raúl Ricardo, Graciela De Antoni, and Angela León Peláez. "Whey permeate fermented with kefir grains shows antifungal effect against Fusarium graminearum." Journal of Dairy Research 83, no. 2 (May 2016): 249–55. http://dx.doi.org/10.1017/s0022029916000121.
Повний текст джерелаАнотація:
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.
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Finkelman, F. D., J. Ohara, D. K. Goroff, J. Smith, N. Villacreses, J. J. Mond, and W. E. Paul. "Production of BSF-1 during an in vivo, T-dependent immune response." Journal of Immunology 137, no. 9 (November 1, 1986): 2878–85. http://dx.doi.org/10.4049/jimmunol.137.9.2878.
Повний текст джерелаАнотація:
Abstract BSF-1, a cytokine produced by some T lymphocyte tumors, has been shown to act with anti-Ig antibodies to stimulate B lymphocyte proliferation, to independently induce resting B lymphocytes to increase their expression of surface Ia antigen, and to induce some activated B lymphocytes to differentiate into IgG1- or IgE-secreting cells. To determine whether BSF-1 might be secreted by normal lymphoid cells in the course of a physiologic immune response, BALB/c mice were injected with an affinity-purified goat antibody to mouse IgD (GaM delta), which induces the generation of a large, polyclonal T-dependent IgG1 response; 4-hr culture supernatants of spleen cells from these mice were prepared, and these supernatants were assayed for BSF-1 activity by analyzing their ability to induce BALB/c nu/nu spleen cells to increase their expression of cell surface Ia in vitro. Culture supernatants of unfractionated spleen cells removed from mice 4 to 8 days after GaM delta antibody injection induced substantial increases in B lymphocyte surface Ia expression; these increases were blocked by a monoclonal anti-BSF-1 antibody. Culture supernatants of spleen cells from untreated BALB/c mice or from untreated or GaM delta antibody-treated BALB/c nu/nu mice induced small to moderate increases in B cell surface Ia expression, and GaM delta antibody itself induced large increases in B cell surface Ia expression; however, these increases were not significantly blocked by a monoclonal anti-BSF-1 antibody. A culture supernatant of T cell-enriched spleen cells from untreated mice induced small increases in B cell surface Ia expression that were inhibited by anti-BSF-1 antibody, as was the larger increase in B cell Ia expression induced by a culture supernatant of T cell-enriched spleen cells from mice sacrificed 3 days after GaM delta injection. On the other hand, T cell-depleted spleen cells from BALB/c mice injected with GaM delta antibody 7 days before sacrifice failed to generate culture supernatants with BSF-1 activity. Supernatants prepared from spleen cells taken from untreated mice or mice treated with GaM delta antibody 1 to 3 days before sacrifice did not block the ability of purified BSF-1 to induce an increase in B cell surface Ia expression, and thus did not contain inhibitors of BSF-1 activity.(ABSTRACT TRUNCATED AT 400 WORDS)
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Pène, J., F. Rousset, F. Brière, I. Chrétien, X. Paliard, J. Banchereau, H. Spits, and J. E. De Vries. "IgE production by normal human B cells induced by alloreactive T cell clones is mediated by IL-4 and suppressed by IFN-gamma." Journal of Immunology 141, no. 4 (August 15, 1988): 1218–24. http://dx.doi.org/10.4049/jimmunol.141.4.1218.
Повний текст джерелаАнотація:
Abstract Seven T cell clones were established from mixed leukocyte cultures in which PBMC from two healthy donors and from one patient suffering from the hyper-IgE syndrome were stimulated by the irradiated EBV-transformed B cell lines JY or UD53. Five of seven T cell clones, after activation by co-cultivation with JY or UD53 cells, induced a low degree of IgE production by normal blood B cells. In one experiment in which the normal B cells could activate the T cell clones directly, IgE production was also observed in the absence of the specific stimulator cells. IgE production was also obtained with supernatants of the T cell clones collected 4 to 5 days after activation by their specific stimulator cells. In addition, the supernatants induced IgG, IgA, and IgM synthesis. All seven clones produced variable concentrations of IL-4 and IFN-gamma. The clones FA-28 and BG-39, which failed to induce IgE synthesis, produced, compared with the other clones tested, relatively high quantities of IFN-gamma (4700 and 2500 pg/ml, respectively). These high levels of IFN-gamma accounted for the lack of induction of IgE synthesis, because in the presence of a polyclonal anti-IFN-gamma antiserum, supernatants of FA-10 and BG-39 induced significant IgE production. In addition, the low degree of IgE production induced by supernatants of two other T cell clones (FA28 and BG24) was 15- and 3-fold enhanced, respectively, in the presence of the anti-IFN-gamma antiserum. IgE synthesis by normal B cells was also induced by rIL-4, indicating that IL-4 present in T cell clone supernatants was responsible for induction of IgE production. This notion was supported by the finding that IgE production induced by supernatant of BG-24 was strongly inhibited by a polyclonal anti-IL-4 antiserum. In contrast, IgG and IgA production induced by supernatant of BG-24 were not significantly affected by the anti-IL-4 antiserum. Only a slight inhibition of IgM synthesis was observed. Collectively, our results indicate that both recombinant and naturally produced IL-4 induce normal human B cells to synthesize IgE. However, final IgE production induced by T cell clone supernatants is the net result of the inducing and suppressive effects of IL-4 and IFN-gamma respectively, that are secreted simultaneously by the T cell clones upon activation.
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Francis, Frédéric, Florent Druart, José Diana Di Mavungu, Marthe De Boevre, Sarah De Saeger, and Frank Delvigne. "Biofilm Mode of Cultivation Leads to an Improvement of the Entomotoxic Patterns of Two Aspergillus Species." Microorganisms 8, no. 5 (May 11, 2020): 705. http://dx.doi.org/10.3390/microorganisms8050705.
Повний текст джерелаАнотація:
Two fungi, i.e., Aspergillus flavus Link and Aspergillus oryzae (Ahlb.) E. Cohn, were cultivated according to two methodologies, namely submerged and biofilm cultures with the primary aim to use their secondary metabolites the supernatant CL50, and CL90 varied between 1.3% (v/v) to 12.7% (v/v) for incubation times from 24 to 72 h. While the A. flavus supernatant entomotoxicity was higher than this of A. oryzae, the biofilm culture application increased the efficiency of the former. Proteomic analysis of the supernatants revealed discrepancies among the two species and modes of cultivation. Furthermore, the secondary metabolite profiles of both Aspergillus cultures were verified. Aspergillic acid, beta-cyclopiazonic acid, cyclopiazonic acid, ferrineospergillin, flavacol, and spermadin A were most predominant. Generally, these secondary metabolites were present in higher concentrations in the supernatants of A. flavus and biofilm cultures. These molecular identifications correlated positively with entomotoxic activity. Noteworthy, the absence of carcinogenic aflatoxins was remarkable, and it will allow further valorization to produce A. flavus to develop potential biopesticides.
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Abdelsalam, Karim, Mrigendra Rajput, Gamal Elmowalid, Jacob Sobraske, Neelu Thakur, Hossam Abdallah, Ahmed A. H. Ali, and Christopher C. L. Chase. "The Effect of Bovine Viral Diarrhea Virus (BVDV) Strains and the Corresponding Infected-Macrophages’ Supernatant on Macrophage Inflammatory Function and Lymphocyte Apoptosis." Viruses 12, no. 7 (June 29, 2020): 701. http://dx.doi.org/10.3390/v12070701.
Повний текст джерелаАнотація:
Bovine viral diarrhea virus (BVDV) is an important viral disease of cattle that causes immune dysfunction. Macrophages are the key cells for the initiation of the innate immunity and play an important role in viral pathogenesis. In this in vitro study, we studied the effect of the supernatant of BVDV-infected macrophage on immune dysfunction. We infected bovine monocyte-derived macrophages (MDM) with high or low virulence strains of BVDV. The supernatant recovered from BVDV-infected MDM was used to examine the functional activity and surface marker expression of normal macrophages as well as lymphocyte apoptosis. Supernatants from the highly virulent 1373-infected MDM reduced phagocytosis, bactericidal activity and downregulated MHC II and CD14 expression of macrophages. Supernatants from 1373-infected MDM induced apoptosis in MDBK cells, lymphocytes or BL-3 cells. By protein electrophoresis, several protein bands were unique for high-virulence, 1373-infected MDM supernatant. There was no significant difference in the apoptosis-related cytokine mRNA (IL-1beta, IL-6 and TNF-a) of infected MDM. These data suggest that BVDV has an indirect negative effect on macrophage functions that is strain-specific. Further studies are required to determine the identity and mechanism of action of these virulence factors present in the supernatant of the infected macrophages.
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Ríos, Diana L., Catalina López, and Jorge U. Carmona. "Platelet-Rich Gel Supernatants Stimulate the Release of Anti-Inflammatory Proteins on Culture Media of Normal Equine Synovial Membrane Explants." Veterinary Medicine International 2015 (2015): 1–9. http://dx.doi.org/10.1155/2015/547052.
Повний текст джерелаАнотація:
The aims were as follows: (1) to evaluate the effects at 48 and 96 h of two concentrations (25 and 50%) of leukocyte and platelet-rich gel (L-PRG) and pure PRG (P-PRG) supernatants on the production/degradation in normal equine synovial membrane explants (SME) of platelet derived growth factor isoform BB, transforming growth factor beta-1, tumor necrosis factor alpha, interleukin (IL-) 4 (IL-4), IL-1 receptor antagonist (IL-1ra), and hyaluronan (HA) synthesis and (2) to correlate these molecules with their respective PRG supernatant treatments. SME from 6 horses were cultured for 96 h with L-PRG and P-PRG supernatants at 25 and 50% concentrations, respectively. SME culture media were changed each 48 h and used for determination by ELISA of the molecules, which were also determined in synovial fluid. 25% L-PRG supernatant produced a sustained release over time of IL-1ra and a gradual release of HA, whereas 50% L-PRG supernatant produced a sustained increase over time of IL-4 and HA. 50% P-PRG supernatant produced an increased and sustained production of IL-1ra and IL-4. The cellular composition and the articular concentration (volume) of a platelet-rich plasma preparation could affect the anti-inflammatory and anabolic joint responses in horses with osteoarthritis.
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Abbood, Wasan, and Alice Krekor. "Study of Hydrophobicity and Autoaggregation of Lactobacillus acidophilus Isolated from vagina." Journal of Biotechnology Research Center 3, no. 2 (June 1, 2009): 29–40. http://dx.doi.org/10.24126/jobrc.2009.3.2.64.
Повний текст джерелаАнотація:
The Hydrophobicity of the seven isolates of L.acidophilus were detected by applying BATH test (Bacterial Adherence To Hydrocarbons) using xylene. The percentage of Hydrophobicity of the isolates ranged between 28-88% and the differences between the rates were significant (P<0.05). There was three hydrophobic isolates. The autoaggregation ability of the three isolates was tested by using the supernatant of MRS and LAPTg media in which the bacteria was cultivated for 24 hr. The results revealed that the three isolates were more aggregative in LAPTg supernatant. The percentage of the aggregation ranged between 70- 83.3% .On the other hand the percentage of the autoaggregation using MRS supernatant ranged between 30-70%. The nature of the surface and secreted factors which are responsible for the autoaggregation were determined by treatment of the bacterial cells and their LAPTg supernatants by proteinase K, lipase and sodium periodate. Results obtained indicated that the two types of these factors were proteins because of the inhibition of the aggregation after either the treatment of cells or the supernatants by proteinase K, and it's resistance to treatment with lipase or sodium periodate.
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Maidana, L. G., J. Gerez, F. Pinho, S. Garcia, and A. P. Bracarense. "Histopathological and ultrastructural findings induced by heat-inactivated Lactobacillus plantarum and the culture supernatant on the intestinal mucosa of piglets: an ex vivo approach." Arquivo Brasileiro de Medicina Veterinária e Zootecnia 71, no. 1 (February 2019): 11–20. http://dx.doi.org/10.1590/1678-4162-10216.
Повний текст джерелаАнотація:
ABSTRACT In the present study, histological, morphometrical and ultrastructural analysis were performed to investigate intestinal mucosa changes in piglets jejunal explants exposed to two concentration of heat-inactivated Lactobacillus plantarum and their respective culture supernatants. Jejunal explants were incubated for 4 hours in DMEM culture medium with a) only culture medium (control group), b) heat-inactivated Lactobacillus plantarum strain1 - LP1 (1.1 x 108CFU/ml), c) heat-inactivated Lactobacillus plantarum strain2 - LP2 (2.0 x 109CFU/ml), d) heat-inactivated Lactobacillus plantarum strain1 culture supernatant (CS1), and e) heat-inactivated Lactobacillus plantarum strain2 culture supernatant (CS2). Explants exposed to heat-inactivated L. plantarum strain 1 and 2 showed multifocal to difuse villi atrophy, villi apical necrosis and enterocyte flattening. Morphological assessment revealed similar results with bacterial adhesion to mucus and intestinal epithelial cells and, morphometric analysis showed a decreased villi height compared to the control group. Alterations in explants treated with the culture supernatant of both strains include mild villi atrophy and mild enterocyte apical necrosis. Morphological assesment reveled numerous well delineated villi and, morphometric analysis showed a significant increase in villi height compared to the control group. In general, exposure to the culture supernatants improved the intestinal morphology.
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Lopez Lopez, C. D., J. O. Jaramillo Polanco, S. Vanner, and D. E. Reed. "A304 FOOD ANTIGEN-STRESS INTERACTION INCREASES PERIPHERAL PAIN SIGNALING IN A MOUSE MODEL OF IBS." Journal of the Canadian Association of Gastroenterology 1, suppl_1 (February 2018): 529. http://dx.doi.org/10.1093/jcag/gwy008.305.
Повний текст джерелаАнотація:
Abstract Background Food and stress are common triggers of symptoms in irritable bowel syndrome (IBS). However, it is unknown whether a specific food antigen alone can induce symptoms such as abdominal pain or if a second trigger, such as stress, is required for a food antigen to increase pain signaling. Aims To determine if a food antigen (ovalbumin) can induce hyperexcitabiity in nociceptive neurons alone and/or in combination with stress. Methods Mice were exposed to water avoidance stress (WAS; 1 hr for 6 days). On day 2, mice were gavaged ovalbumin (20mg) daily after completing WAS (WAS/OVA). These were compared to 3 groups: no WAS and no ovalbumin (control); WAS but no ovalbumin (WAS); and ovalbumin (OVA) alone. On day 8, WAS, OVA and WAS/OVA mice were given ovalbumin (50 μg) subcutaneously; 3 hours later euthanized. Colons were removed and incubated with ovalbumin (100 μg/ml) for 4 hrs. Supernatants were collected and DRG neurons from control mice were incubated overnight with supernatants from each of the four groups. Changes in neuronal excitability were examined by measuring the rheobase (minimal current to evoke an action potential) using perforated patch clamp recording techniques. Stress effects on intestinal permeability in the ileum and colon were assessed in Ussing chambers. One way ANOVA and Bonferroni post hoc test or unpaired t test were used to analyze the data. Results Incubation with supernatants obtained from WAS/OVA mice evoked hyperexcitability in DRG neurons compared to incubation with control supernatant (rheobase: control = 82 ± 10 pA vs WAS/OVA = 56 ± 4 pA, p < 0.05). Similarly, WAS/OVA supernatant decreased the rheobase compared to both OVA mice (OVA = 80 ± 8 pA, p<0.05) and WAS mice (WAS = 81 ± 9 pA, p<0.05). The effect of supernatants from OVA mice and WAS mice did not differ from controls. In a separate series of experiments, the PAR2 antagonist GB83 inhibited the effect of WAS/OVA supernatant (p < 0.01). There was no effect of GB83 on WAS or OVA supernatants. Stress decreased tissue resistance in ileum (p<0.05) but not in colon. Conclusions The food antigen ovalbumin induced hyperexcitability in DRG nociceptive neurons only when combined with stress. This action was PAR2 dependent suggesting a role of tissue proteases. Stress may cause a loss of oral tolerance to ovalbumin by increasing mucosal permeability in the small intestine. The interaction of food antigens and stress may be a mechanism of meal induced increase in abdominal pain in IBS. Funding Agencies CIHR
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Young, Judy C., Xin Yu, Allen Nguyen, Catherine Kung, Wai Lee Wong, Laura DeForge, and Jane Grogan. "Lymphotoxin-alpha beta heterotrimers are shed from the surface of activated human lymphocytes, circulate in serum, and are elevated in synovial fluid of rheumatoid arthritis patients (38.22)." Journal of Immunology 182, no. 1_Supplement (April 1, 2009): 38.22. http://dx.doi.org/10.4049/jimmunol.182.supp.38.22.
Повний текст джерелаАнотація:
Abstract Lymphotoxin, a trimeric cytokine in the tumor necrosis family, is expressed by activated T, B, and NK cells and is involved in inflammatory response signaling and secondary lymphoid organ architecture. Using a specific assay for human LTαβ which detects both LTα1β2 and LTα2β1 trimers but not LTα3, we show here that LTαβ is detected in culture supernatants of 293 cells transfected with human LTα and LTβ constructs, and in supernatants of primary activated human T lymphocytes. The levels of soluble LTαβ detected in culture supernatants of human CD4+ Th1 polarized lymphocytes treated with an inhibitor of ADAM17 (TACE) protease, were reduced 3-6 fold compared to supernatants from untreated cells. This suggests that LTαβ cleavage is carried out by TACE. A 26 KDa MW form of LTβ was detected in activated human T cell culture supernatant by Western blot analysis using an anti-LTβ-specific antibody. Soluble LTαβ was detected in serum from normal human donors and rheumatoid arthritis (RA) patients at levels of approximately 100 pg/mL. Soluble LTαβ was also detected in synovial fluid from RA patients at significantly higher levels than in synovial fluid from osteoarthritis patients. Cleavage of LTαβ trimers from the cell membrane and release into circulation has not been previously described. Soluble LTαβ may play a role as an inflammatory mediator of autoimmune disease. Research supported by Genentech Inc., S. San Francisco, CA
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Cheng, Ling, Weibiao Cao, Claudio Fiocchi, Jose Behar, Piero Biancani, and Karen M. Harnett. "In vitro model of acute esophagitis in the cat." American Journal of Physiology-Gastrointestinal and Liver Physiology 289, no. 5 (November 2005): G860—G869. http://dx.doi.org/10.1152/ajpgi.00260.2005.
Повний текст джерелаАнотація:
We have shown that IL-1β and IL-6, possibly originating from the mucosa in response to injury, inhibit neurally mediated contraction of esophageal circular muscle but do not affect ACh-induced contraction, reproducing the effect of experimental esophagitis on esophageal contraction. To examine the interaction of mucosa and circular muscle in inflammation, we examined the effect of HCl on in vitro esophageal mucosa and circular muscle. Circular muscle strips, when directly exposed to HCl, contracted normally. However, when circular muscle strips were exposed to supernatants of mucosa incubated in HCl (2–3 h, pH 5.8), contraction decreased, and the inhibition was partially reversed by an IL-6 antibody. Supernatants from the mucosa of animals with in vivo-induced acute esophagitis (AE) similarly reduced contraction. IL-6 levels were higher in mucosal tissue from AE animals than in control mucosa and in AE mucosa supernatants than in normal mucosa supernatants. IL-6 levels increased significantly in normal mucosa and supernatants in response to HCl, suggesting increased production and release of IL-6 by the mucosa. IL-6 increased H2O2 levels in the circular muscle layer but not in mucosa. Exposure of the mucosa to HCl caused IL-1β to increase only in the mucosa and not in the supernatant. These data suggest that HCl-induced damage occurs first in the mucosa, leading to the production of IL-1β and IL-6 but not H2O2. IL-1β appears to remain in the mucosa. In contrast, IL-6 is produced and released by the mucosa, eventually resulting in the production of H2O2 by the circular muscle, with this affecting circular muscle contraction.
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Glogowski, J., I. Babiak, K. Goryczko, and S. Dobosz. "Activity of aspartate aminotransferase and acid phosphatase in cryopreserved trout sperm." Reproduction, Fertility and Development 8, no. 8 (1996): 1179. http://dx.doi.org/10.1071/rd9961179.
Повний текст джерелаАнотація:
Milt of brown, rainbow and brook trout was cryopreserved. Activity of aspartate aminotransferase (AspAT) and acid phosphatase was assayed both in supernatants and in spermatozoa obtained from thawed sperm samples; additionally, post-thaw motility was evaluated. Enzyme activities differed according to fish species and were strongly affected by the type of cryoprotectant used. The activity in supernatants was usually higher than that in spermatozoa because of protein leakage from injured cells. AspAT activity in cryopreserved spermatozoa correlated positively with fertilization success in all three species. There was a negative correlation between activity of extracellular (supernatant) AspAT and fertilization rates in variants with dimethyl sulfoxide and dimethylacetamide-based extenders. The motility of thawed sperm, determined microscopically, provided some information on the cryopreservation efficiency of trout milt.
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Arbizu, Shirley, Giuliana Noratto, and Susanne Talcott. "Assessment of Whey Functional Ingredients in the Modulation of Fecal Bacteria from Donors with Chronic Gastrointestinal Disease In Vitro." Current Developments in Nutrition 4, Supplement_2 (May 29, 2020): 736. http://dx.doi.org/10.1093/cdn/nzaa052_005.
Повний текст джерелаАнотація:
Abstract Objectives To evaluate whey functional ingredients (WFI) as source of non-digestible nutrients for fecal bacteria from donors with intestinal bowel disease (IBD) in vitro. Methods The WFI whey protein isolate (WPI), glycomacropeptide (GMP) and a galacto-oligosaccharide rich whey protein concentrate (GOS-W), and peptone (control) were subjected to in vitro digestion (IVD) and freeze dried to be used in fecal culture medium. Fecal de-identified samples from 10 healthy and 9 mild-moderate IBD subjects were subjected to in vitro fecal fermentation for 24 h. Fecal bacteria and culture supernatants were analyzed using standard analytical procedures to quantify bacteria relative abundance, and metabolites in culture supernatants. Short chain fatty acids were quantified in fecal supernatants by HPLC analysis. HT29-MTX intestinal cells were treated with sterile-filtered fecal culture supernatants (2.5% v/v) to assess production of reactive oxygen species (ROS) using 10 μM of 2′7′ dichlorodihydrofluorescein diacetate (H2DCFDA) reagent. Results Among the WFI tested, WPI tended to modulate the relative abundance of bacteria that have been reported to decrease during IBD conditions such as R. hominis, R. intestinalis and R. torques. Metabolites in fecal culture supernatants showed that propionic acid concentrations in IBD controls were higher than healthy controls and WFI fermentations decreased those levels making them similar to the healthy controls. In contrast, the concentration of lactic acid tended to be higher in the GOS-W fecal culture supernatant, but only reached significance (P < 0.05) when compared to GMP-supplemented medium. No differences in acetic acid and butyric acid concentrations were found between IBD controls and WFI treatments. Results also showed that WPI, GOS-W and GMP fecal culture supernatants prevented ROS production in HT29-MTX cells when compared to their respective IBD-controls. Conclusions WPI favorably modulated the relative abundance of bacteria relevant in IBD while all WFI metabolites produced after in vitro fecal fermentation mitigated oxidative stress. These findings suggest the potential of WFI to moderate adverse conditions associated with IBD. Funding Sources Build Dairy Program.
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Sixt, S. U., R. Alami, J. Hakenbeck, M. Adamzik, A. Kloß, U. Costabel, P. R. Jungblut, B. Dahlmann, and J. Peters. "Distinct Proteasome Subpopulations in the Alveolar Space of Patients with the Acute Respiratory Distress Syndrome." Mediators of Inflammation 2012 (2012): 1–9. http://dx.doi.org/10.1155/2012/204250.
Повний текст джерелаАнотація:
There is increasing evidence that proteasomes have a biological role in the extracellular alveolar space, but inflammation could change their composition. We tested whether immunoproteasome protein-containing subpopulations are present in the alveolar space of patients with lung inflammation evoking the acute respiratory distress syndrome (ARDS). Bronchoalveolar lavage (BAL) supernatants and cell pellet lysate from ARDS patients (n=28) and healthy subjects (n=10) were analyzed for the presence of immunoproteasome proteins (LMP2 and LMP7) and proteasome subtypes by western blot, chromatographic purification, and 2D-dimensional gelelectrophoresis. In all ARDS patients but not in healthy subjects LMP7 and LMP2 were observed in BAL supernatants. Proteasomes purified from pooled ARDS BAL supernatant showed an altered enzyme activity ratio. Chromatography revealed a distinct pattern with 7 proteasome subtype peaks in BAL supernatant of ARDS patients that differed from healthy subjects. Total proteasome concentration in BAL supernatant was increased in ARDS (971 ng/mL±1116 versus59±25;P<0.001), and all fluorogenic substrates were hydrolyzed, albeit to a lesser extent, with inhibition by epoxomicin (P=0.0001). Thus, we identified for the first time immunoproteasome proteins and a distinct proteasomal subtype pattern in the alveolar space of ARDS patients, presumably in response to inflammation.
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Holcombe, Lucy J., Gordon McAlester, Carol A. Munro, Brice Enjalbert, Alistair J. P. Brown, Neil A. R. Gow, Chen Ding, Geraldine Butler, Fergal O'Gara, and John P. Morrissey. "Pseudomonas aeruginosa secreted factors impair biofilm development in Candida albicans." Microbiology 156, no. 5 (May 1, 2010): 1476–86. http://dx.doi.org/10.1099/mic.0.037549-0.
Повний текст джерелаАнотація:
Signal-mediated interactions between the human opportunistic pathogens Pseudomonas aeruginosa and Candida albicans affect virulence traits in both organisms. Phenotypic studies revealed that bacterial supernatant from four P. aeruginosa strains strongly reduced the ability of C. albicans to form biofilms on silicone. This was largely a consequence of inhibition of biofilm maturation, a phenomenon also observed with supernatant prepared from non-clinical bacterial species. The effects of supernatant on biofilm formation were not mediated via interference with the yeast–hyphal morphological switch and occurred regardless of the level of homoserine lactone (HSL) produced, indicating that the effect is HSL-independent. A transcriptome analysis to dissect the effects of the P. aeruginosa supernatants on gene expression in the early stages of C. albicans biofilm formation identified 238 genes that exhibited reproducible changes in expression in response to all four supernatants. In particular, there was a strong increase in the expression of genes related to drug or toxin efflux and a decrease in expression of genes associated with adhesion and biofilm formation. Furthermore, expression of YWP1, which encodes a protein known to inhibit biofilm formation, was significantly increased. Biofilm formation is a key aspect of C. albicans infections, therefore the capacity of P. aeruginosa to antagonize this has clear biomedical implications.
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Menu, E., A. Bensussan, V. David, and G. Chaouat. "Effects of placental supernatants (human) and choriocarcinoma supernatants on human T cell clones." Journal of Reproductive Immunology 15 (July 1989): 141. http://dx.doi.org/10.1016/0165-0378(89)90288-x.
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Rivas, J. M., and S. E. Ullrich. "Systemic suppression of delayed-type hypersensitivity by supernatants from UV-irradiated keratinocytes. An essential role for keratinocyte-derived IL-10." Journal of Immunology 149, no. 12 (December 15, 1992): 3865–71. http://dx.doi.org/10.4049/jimmunol.149.12.3865.
Повний текст джерелаАнотація:
Abstract Exposing murine keratinocyte cultures to UV radiation causes the release of a suppressive cytokine that mimics the immunosuppressive effects of total-body UV exposure. Injecting supernatants from UV-irradiated keratinocyte cultures into mice inhibits their ability to generate a delayed-type hypersensitivity reaction against allogeneic histocompatibility Ag, and spleen cells from mice injected with supernatant do not respond to alloantigen in the in vitro MLR. A unique feature of the immunosuppression induced by either total-body UV-exposure or injecting the suppressive cytokine from UV-irradiated keratinocytes is the selectivity of suppression. Although cellular immune reactions such as delayed-type hypersensitivity are suppressed antibody production is unaffected. Because the selective nature to the UV-induced immunosuppression is similar to the biologic activity of IL-10, we examined the hypothesis that UV exposure of keratinocytes causes the release of IL-10. Keratinocyte monolayers were exposed to UV radiation and at specific times after exposure mRNA was isolated or the culture supernatant from the cells was collected. IL-10 mRNA expression was enhanced in UV-irradiated keratinocytes. The secretion of IL-10 by the irradiated keratinocytes was determined by Western blot analysis. A band reactive with anti-IL-10 mAb was found in supernatants from the UV-irradiated but not the mock-irradiated cells. IL-10 biologic activity was determined by the ability of the supernatants from the UV-irradiated keratinocytes to suppress IFN-gamma production by Ag-activated Th 1 cell clones. Anti-IL-10 mAb neutralized the ability of supernatants from UV-irradiated keratinocytes to suppress the induction of delayed-type hypersensitivity in vivo. Furthermore, injecting UV-irradiated mice with antibodies against IL-10 partially inhibited in vivo immunosuppression. These data indicate that activated keratinocytes are capable of secreting IL-10 and suggest that the release of IL-10 by UV-irradiated keratinocytes plays an essential role in the induction of systemic immunosuppression after total-body UV exposure.
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Gajewski, T. F., E. Goldwasser, and F. W. Fitch. "Anti-proliferative effect of IFN-gamma in immune regulation. II. IFN-gamma inhibits the proliferation of murine bone marrow cells stimulated with IL-3, IL-4, or granulocyte-macrophage colony-stimulating factor." Journal of Immunology 141, no. 8 (October 15, 1988): 2635–42. http://dx.doi.org/10.4049/jimmunol.141.8.2635.
Повний текст джерелаАнотація:
Abstract A biphasic dose response curve was observed when the bone marrow-derived cell line FDCP1, used as an indicator line for IL-3 bioassays, was exposed to supernatants from some activated T cell clones but not others. The active component which inhibited proliferation at the higher supernatant concentrations appeared to be IFN-gamma, based on the following observations. 1) Only those culture supernatants which contained IFN-gamma gave a biphasic dose response curve; 2) with these supernatants, an anti-IFN-gamma mAb augmented the proliferation of FDCP1 cells at the higher supernatant concentrations; and 3) rIFN-gamma profoundly inhibited the proliferation of FDCP1 cells stimulated with rIL-3 or rIL-4. rTNF-alpha inhibited FDCP1 proliferation only to a modest extent, yet the combination of rTNF-alpha + rIFN-gamma provided greater inhibition than each agent alone. The proliferation of a second bone marrow-derived cell line, DA1, was not inhibited by rIFN-gamma or rIFN-gamma + rTNF-alpha when stimulated with rIL-3 or recombinant granulocyte-macrophage colony-stimulating factor (rGM-CSF). Fresh bone marrow cells also showed a suboptimal proliferative response when stimulated with T cell supernatants containing IFN-gamma, and this response was augmented considerably upon the addition of anti-IFN-gamma mAb. Bone marrow cell proliferation was observed upon exposure to rIL-3, rIL-4, or rGM-CSF, and these responses were inhibited by rIFN-gamma; rTNF-alpha also produced a synergistic effect with these cells. Bone marrow cell colony formation stimulated by rIL-3 or rGM-CSF also was inhibited by rIFN-gamma. Colony formation in bone marrow cell cultures was not observed in response to rIL-4. Collectively, these results suggest that Th1 cells, which in addition to IL-3 and GM-CSF also produce IFN-gamma, may regulate hemopoietic cell proliferation and colony formation differently from the way Th2 cells do, which do not produce IFN-gamma.
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Santoli, D., D. J. Tweardy, D. Ferrero, B. L. Kreider, and G. Rovera. "A suppressor lymphokine produced by human T leukemia cell lines. Partial characterization and spectrum of activity against normal and malignant hemopoietic cells." Journal of Experimental Medicine 163, no. 1 (January 1, 1986): 18–40. http://dx.doi.org/10.1084/jem.163.1.18.
Повний текст джерелаАнотація:
Human T leukemia cell lines spontaneously release into their medium a suppressor lymphokine, T leukemia-derived suppressor lymphokine (TLSL), able to inhibit proliferation, DNA synthesis, and colony formation in a variety of malignant hemopoietic cell lines, as well as in normal myelomonocytic progenitor cells from bone marrow and peripheral blood. Titration curves indicated that the inhibitory activity in the crude supernatant preparations ranged from 10(-3)-10(-9): the supernatants from CCRF/CEM, HUT-78, and MOLT-4 cell lines were the most active, those from HPB-ALL, JM, and CCRF/HSB2 displayed an intermediate activity, and the Jurkat supernatant was the least active. Target cell lines of B cell origin (Burkitt lymphomas) were more sensitive than granulocytic, monocytic, erythroid, and T cell lines. Partial purification by ammonium sulfate precipitation and column chromatography demonstrated that TLSL is a protein with an Mr of 88,000, as determined by gel filtration. A high Mr form (greater than 300,000) was produced in serum-free medium by one of the most active producer cell lines (CCRF/CEM), and appeared to be an aggregate of the 88,000 Mr form. Neither the partially purified fractions obtained nor the crude supernatant preparations displayed antiviral activity or contained interleukin 2. Unlike lymphotoxin and tumor necrosis factor, TLSL is cytostatic: maximal inhibition of proliferation was observed 4-5 d after addition of crude supernatant to the target cells, and was not accompanied by a significant loss in cell viability. The antiproliferative capacity of TLSL was manifested both in suspension and methylcellulose cultures. Treated target cells accumulated either in the G1 or in the S phase of the cell cycle. The effect of TLSL on the target cells is irreversible: even brief (1 h) incubation of sensitive cells with TLSL resulted in inhibition of proliferation measured 5 d later. Although TLSL is produced by leukemic T cell lines, this lymphokine inhibits proliferation of normal peripheral blood T cells in response to mitogens or alloantigens: T lymphocyte activation was inhibited by all of the T cell supernatants tested. In contrast, when T cell lines were used as targets, no inhibition of proliferation was detected with two exceptions: the low producer Jurkat cell line was sensitive to all the T cell-derived supernatants, and the intermediate producer CCRF/HSB2 cell line was sensitive only to the three most active supernatants, CCRF/CEM, MOLT-4, and HUT-78. The possible significance of TLSL and its relationship with other suppressor lymphokines previously described in other systems is discussed.
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Yoshimori, Mayumi, Miwako Nishio, Ayaka Ohashi, Megumi Tateishi, Ayaka Mimura, Naomi Wada, Minori Saito, Norio Shimizu, Ken-Ichi Imadome та Ayako Arai. "Interferon-γ Produced by EBV-Positive Neoplastic NK-Cells Induces Differentiation into Macrophages and Procoagulant Activity of Monocytes, Which Leads to HLH". Cancers 13, № 20 (12 жовтня 2021): 5097. http://dx.doi.org/10.3390/cancers13205097.
Повний текст джерелаАнотація:
Epstein–Barr virus (EBV)-positive T- or NK-cell neoplasms show progressive systemic inflammation and abnormal blood coagulation causing hemophagocytic lymphohistiocytosis (HLH). It was reported that inflammatory cytokines were produced and secreted by EBV-positive neoplastic T- or NK-cells. These cytokines can induce the differentiation of monocytes into macrophages leading to HLH. To clarify which products of EBV-positive neoplastic T- or NK-cells have effects on monocytes, we performed a co-culture assay of monocytes with the supernatants of EBV-positive T- or NK-cell lines. The expression of differentiation markers, the phagocytosis ability, and the mRNA expression of the inflammatory cytokines of THP-1, a monocytic cell line, clearly increased after culturing with the supernatants from EBV-NK-cell lines. Co-culturing with the supernatants promoted the expression of CD80 and CD206 as well as M1 and M2 macrophage markers in human monocytes. Co-culturing with the supernatants of EBV-NK-cell lines significantly enhanced the procoagulant activity and the tissue factor expression of monocytes. Interferon (IFN)-γ was elevated extremely not only in the supernatant of EBV-NK-cell lines but also in the plasma of EBV-positive NK-cell neoplasms patients accompanying HLH. Finally, we confirmed that IFN-γ directly enhanced the differentiation into M1-like macrophages and the procoagulant activity of monocytes. Our findings suggest that IFN-γ may potentially serve as a therapeutic target to regulate HLH in EBV-positive NK-cell neoplasms.
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Chen, Jianming, and Bruce A. McClane. "Role of the Agr-Like Quorum-Sensing System in Regulating Toxin Production by Clostridium perfringens Type B Strains CN1793 and CN1795." Infection and Immunity 80, no. 9 (June 11, 2012): 3008–17. http://dx.doi.org/10.1128/iai.00438-12.
Повний текст джерелаАнотація:
ABSTRACTClostridium perfringenstype B causes enteritis and enterotoxemia in domestic animals. By definition, these bacteria must produce alpha toxin (CPA), beta toxin (CPB) and epsilon toxin (ETX) although most type B strains also produce perfringolysin O (PFO) and beta2 toxin (CPB2). A recently identified Agr-like quorum-sensing (QS) system inC. perfringenscontrols all toxin production by surveyed type A, C, and D strains, but whether this QS is involved in regulating toxin production by type B strains has not been explored. Therefore, the current study introducedagrBnull mutations into type B strains CN1795 and CN1793. Both type BagrBnull mutants exhibited reduced levels of CPB, PFO, and CPA in their culture supernatants, and this effect was reversible by complementation. The reduced presence of CPB in culture supernatant involved decreasedcpbtranscription. In contrast, theagrBnull mutants of both type B strains retained wild-type production levels of ETX and CPB2. In a Caco-2 cell model of enteritis, culture supernatants of the type BagrBnull mutants were less cytotoxic than supernatants of their wild-type parents. However, in an MDCK cellin vitromodel for enterotoxemic effects, supernatants from theagrBnull mutants or wild-type parents were equally cytotoxic after trypsin activation. Coupling these and previous results, it is now evident that strain-dependent variations exist in Agr-like QS system regulation ofC. perfringenstoxin production. The cell culture results further support a role for trypsin in determining which toxins contribute to disease involving type B strains.
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Varina, Maureen, Steven M. Denkin, Andrew M. Staroscik, and David R. Nelson. "Identification and Characterization of Epp, the Secreted Processing Protease for the Vibrio anguillarum EmpA Metalloprotease." Journal of Bacteriology 190, no. 20 (August 8, 2008): 6589–97. http://dx.doi.org/10.1128/jb.00535-08.
Повний текст джерелаАнотація:
ABSTRACT The zinc metalloprotease EmpA is a virulence factor for the fish pathogen Vibrio anguillarum. Previous studies demonstrated that EmpA is secreted as a 46-kDa proenzyme that is activated extracellularly by the removal of an ∼10-kDa propeptide. We hypothesized that a specific protease is responsible for processing secreted pro-EmpA into mature EmpA. To identify the protease responsible for processing pro-EmpA, a minitransposon mutagenesis (using mini-Tn10Km) clone bank of V. anguillarum was screened for reduced protease activity due to insertions in undescribed genes. One mutant with reduced protease activity was identified. The region containing the mini-Tn10Km was cloned, sequenced, and found to contain epp, an open reading frame encoding a putative protease. Further characterization of epp was done using strain M101, created by single-crossover insertional mutagenesis. Protease activity was absent in M101 cultures even when empA protease activity was induced by salmon gastrointestinal mucus. When the epp mutation was complemented with a wild-type copy of epp (M102), protease activity was restored. Western blot analysis of sterile filtered culture supernatants from wild-type (M93Sm) cells, M101 cells, and M102 cells revealed that only pro-EmpA was present in M101supernatants; both pro-EmpA and mature EmpA were detected in M93Sm and M102 supernatants. When sterile filtered culture supernatants from the empA mutant strain (M99) and M101 were mixed, protease activity was restored. Western blot analysis revealed that pro-EmpA in M101 culture supernatant was processed to mature EmpA only after mixing with M99 culture supernatant. These data show that Epp is the EmpA-processing protease.