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51.
In the present study, the cultural conditions for exopolysaccharide (EPS) production from probiotic yeast Lipomyces starkeyi VIT-MN03 were optimized using response surface methodology (RSM) to maximize the yield of EPS. Interactions among the various factors viz. sucrose concentration (1–3 g%), NaCl concentration (2–4 g%), pH (3–5), temperature (20–30 °C), and incubation period (20–40 days) during EPS production were studied using Box-Behnken design (BBD). The EPS was purified and characterized using various instrumental analyses. The properties like adhesion, antioxidant, biosurfactant, cholesterol removal, and binding ability to mutagens were also tested for EPS produced. Sixfold increase in EPS production (4.87 g L−1) by L. starkeyi VIT-MN03 was noted under optimized condition. EPS showed a high viscosity (1.8 Pa S−1) and good shear-thinning properties. Instrumental analysis showed that EPS was heteropolysaccharide composed of glucan, mannan, and rhamnan. Lipomyces starkeyi VIT-MN03 exhibited good self-adhesion (95%) and co-aggregation ability (93%). Adhesion efficiency for yeast inoculum containing 5.5 × 107 CFU mL−1 per 9.2 cm2 of Caco-2 cell (colorectal adenocarcinoma) was noted. The probiotic EPS displayed strong antioxidant ability to scavenge hydroxyl radical and DPPH by 58% and 71% respectively. In addition, biosurfactant activity (86%) and cholesterol removal (90%) ability of probiotic EPS was also tested. EPS bound cells of L. starkeyi VIT-MN03 showed good binding ability to mutagens. These results support the effectiveness of using RSM for maximum EPS production. To the best of our knowledge, this is the first report on optimization of EPS production by probiotic yeast. 相似文献
52.
Iskra Tuero Venkatramanan Mohanram Thomas Musich Leia Miller Diego A. Vargas-Inchaustegui Thorsten Demberg David Venzon Irene Kalisz V. S. Kalyanaraman Ranajit Pal Maria Grazia Ferrari Celia LaBranche David C. Montefiori Mangala Rao Monica Vaccari Genoveffa Franchini Susan W. Barnett Marjorie Robert-Guroff 《PLoS pathogens》2015,11(8)
Many viral infections, including HIV, exhibit sex-based pathogenic differences. However, few studies have examined vaccine-related sex differences. We compared immunogenicity and protective efficacy of monomeric SIV gp120 with oligomeric SIV gp140 in a pre-clinical rhesus macaque study and explored a subsequent sex bias in vaccine outcome. Each immunization group (16 females, 8 males) was primed twice mucosally with replication-competent Ad-recombinants encoding SIVsmH4
env/rev, SIV239
gag and SIV239
nefΔ1–13 and boosted twice intramuscularly with SIVmac239 monomeric gp120 or oligomeric gp140 in MF59 adjuvant. Controls (7 females, 5 males) received empty Ad and MF59. Up to 9 weekly intrarectal challenges with low-dose SIVmac251 were administered until macaques became infected. We assessed vaccine-induced binding, neutralizing, and non-neutralizing antibodies, Env-specific memory B cells and plasmablasts/plasma cells (PB/PC) in bone marrow and rectal tissue, mucosal Env-specific antibodies, and Env-specific T-cells. Post-challenge, only one macaque (gp140-immunized) remained uninfected. However, SIV acquisition was significantly delayed in vaccinated females but not males, correlated with Env-specific IgA in rectal secretions, rectal Env-specific memory B cells, and PC in rectal tissue. These results extend previous correlations of mucosal antibodies and memory B cells with protective efficacy. The gp140 regimen was more immunogenic, stimulating elevated gp140 and cyclic V2 binding antibodies, ADCC and ADCP activities, bone marrow Env-specific PB/PC, and rectal gp140-specific IgG. However, immunization with gp120, the form of envelope immunogen used in RV144, the only vaccine trial to show some efficacy, provided more significant acquisition delay. Further over 40 weeks of follow-up, no gp120 immunized macaques met euthanasia criteria in contrast to 7 gp140-immunized and 2 control animals. Although males had higher binding antibodies than females, ADCC and ADCP activities were similar. The complex challenge outcomes may reflect differences in IgG subtypes, Fc glycosylation, Fc-R polymorphisms, and/or the microbiome, key areas for future studies. This first demonstration of a sex-difference in SIV vaccine-induced protection emphasizes the need for sex-balancing in vaccine trials. Our results highlight the importance of mucosal immunity and memory B cells at the SIV exposure site for protection. 相似文献
53.
54.
Beck Z Karasavvas N Tong J Matyas GR Rao M Alving CR 《Biochemical and biophysical research communications》2007,354(3):747-751
The binding characteristics of two monoclonal antibodies (mAb) to phosphatidylinositol-4-phosphate (PIP) were examined: a murine IgM mAb to PIP; and a human IgG mAb (4E10) that binds both to HIV-1 envelope protein and also to neutral and anionic phospholipids, including PIP. Binding of each mAb to pure PIP was inhibited by Ca(2+) as determined by ELISA. When studied by surface plasmon resonance, liposomes containing PIP could be stripped (i.e., removed) by either Ca(2+) or phosphorylated haptens after binding of the liposomes to the murine anti-PIP antibody attached to a BIAcore chip. In contrast, the binding of liposomal PIP to 4E10 was irreversible and could not be stripped. We therefore conclude that Ca(2+) and phosphate can modulate the initial binding of both types of antibodies to PIP. However, 4E10 binds to liposomal PIP in a two-stage process involving first Ca(2+)-modulated binding to the PIP polar headgroup, followed by irreversible binding to liposomal hydrophobic groups. 相似文献
55.
Immunization with DNA through the skin 总被引:8,自引:0,他引:8
The skin has evolved as a barrier to prevent external agents, including pathogens, from entering the body. It has a complex and efficient immune surveillance system, which includes Langerhans cells and dendritic cells. By targeting the body's natural defense system, skin-DNA immunization attempts to produce an efficient immune response. Nucleic acid vaccines provide DNA for protein expression in a variety of cells, including keratinocytes, Langerhans cells, and dendritic cells, which are located in the two main areas of the skin, the epidermis (the most superficial layer) and the dermis. After maturation, Langerhans cells and dermal dendritic cells can migrate to local lymph nodes where presentation of antigens to T cells can occur and thus start a variety of immunologic responses. Dermal immunization methods described in this article target the epidermis, the dermis, or both and include: (a) stripping; (b) chemical modification; (c) trans-epidermal immunization (transcutaneous immunization or non-invasive vaccination of the skin); (d) gene gun technology; (e) electroporation; (f) intradermal injections; and (g) microseeding. These techniques all require the removal of hair, the circumvention or modification of the stratum corneum layer of the epidermis, and the addition of DNA or amplification of DNA signal. As the biology of the skin and the mechanisms of DNA vaccination are elucidated, these skin immunization techniques will be optimized. With refinement, skin-DNA immunization will achieve the goal of producing a reliable and efficacious immune response to a variety of pathogens. 相似文献
56.
Morin Hydrate Mitigates Cisplatin‐Induced Renal and Hepatic Injury by Impeding Oxidative/Nitrosative Stress and Inflammation in Mice
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Athira K V Rajaram Mohanrao Madhana Eshvendar Reddy Kasala Pavan Kumar Samudrala Mangala Lahkar Ranadeep Gogoi 《Journal of biochemical and molecular toxicology》2016,30(12):571-579
Cisplatin is a widely used chemotherapeutic drug; however, it induces damage on kidney and liver at clinically effective higher doses. Morin hydrate possesses antioxidant, anti‐inflammatory, and anticancer properties. Therefore, we aimed to investigate the effects of morin hydrate (50 and 100 mg/kg, orally) against the renohepatic toxicity induced by a high dose of cisplatin (20 mg/kg, intraperitoneally). Renal and hepatic function, oxidative/nitrosative stress, and inflammatory markers along with histopathology were evaluated. Morin hydrate ameliorated cisplatin‐induced renohepatic toxicity significantly at 100 mg/kg as evidenced from the significant reversal of cisplatin‐induced body weight loss, mortality, functional and structural alterations of kidney, and liver. The protective role offered by morin hydrate against cisplatin‐induced renohepatic toxicity is by virtue of its free radical scavenging property, thereby abating the depletion of cellular antioxidant defense components and through modulation of inflammatory cytokines. We speculate morin hydrate as a protective candidate against renohepatic toxicity of cisplatin. 相似文献
57.
Plants produce a large number of cellulases that are either secreted or anchored in the plasma membrane where they likely function in various aspects of cellulose synthesis, modification and degradation during plant growth and development. Very few of these enzymes have been characterized in any detail, however. Here we attempted to produce two Arabidopsis modular cellulases, which contain a catalytic domain belonging to glycoside hydrolase family 9 (GH9) and a carbohydrate binding module (CBM), in the yeast Pichia pastoris. Neither of the intact modular enzymes was detectably produced, although the independently expressed GH9 catalytic domain of one enzyme was secreted when the protein was expressed at low temperature. Expression of intact and truncated cellulases at the standard temperature caused extensive cell lysis, with release of high concentrations of endogenous proteins into the culture medium. Cell lysis appeared to result from misfolding of cellulase proteins within the Pichia secretory pathway. The toxicity of these misfolded cellulases potentially could be exploited to derive host strains with enhanced capability to fold recombinant secretory proteins. 相似文献
58.
Susan Zolla-Pazner Allan C. deCamp Timothy Cardozo Nicos Karasavvas Raphael Gottardo Constance Williams Daryl E. Morris Georgia Tomaras Mangala Rao Erik Billings Phillip Berman Xiaoying Shen Charla Andrews Robert J. O'Connell Viseth Ngauy Sorachai Nitayaphan Mark de Souza Bette Korber Richard Koup Robert T. Bailer John R. Mascola Abraham Pinter David Montefiori Barton F. Haynes Merlin L. Robb Supachai Rerks-Ngarm Nelson L. Michael Peter B. Gilbert Jerome H. Kim 《PloS one》2013,8(1)
The RV144 clinical trial of a prime/boost immunizing regimen using recombinant canary pox (ALVAC-HIV) and two gp120 proteins (AIDSVAX B and E) was previously shown to have a 31.2% efficacy rate. Plasma specimens from vaccine and placebo recipients were used in an extensive set of assays to identify correlates of HIV-1 infection risk. Of six primary variables that were studied, only one displayed a significant inverse correlation with risk of infection: the antibody (Ab) response to a fusion protein containing the V1 and V2 regions of gp120 (gp70-V1V2). This finding prompted a thorough examination of the results generated with the complete panel of 13 assays measuring various V2 Abs in the stored plasma used in the initial pilot studies and those used in the subsequent case-control study. The studies revealed that the ALVAC-HIV/AIDSVAX vaccine induced V2-specific Abs that cross-react with multiple HIV-1 subgroups and recognize both conformational and linear epitopes. The conformational epitope was present on gp70-V1V2, while the predominant linear V2 epitope mapped to residues 165–178, immediately N-terminal to the putative α4β7 binding motif in the mid-loop region of V2. Odds ratios (ORs) were calculated to compare the risk of infection with data from 12 V2 assays, and in 11 of these, the ORs were ≤1, reaching statistical significance for two of the variables: Ab responses to gp70-V1V2 and to overlapping V2 linear peptides. It remains to be determined whether anti-V2 Ab responses were directly responsible for the reduced infection rate in RV144 and whether anti-V2 Abs will prove to be important with other candidate HIV vaccines that show efficacy, however, the results support continued dissection of Ab responses to the V2 region which may illuminate mechanisms of protection from HIV-1 infection and may facilitate the development of an effective HIV-1 vaccine. 相似文献
59.
Mangala Srinivas Philipp Boehm-Sturm Markus Aswendt Eberhard D. Pracht Carl G. Figdor I. Jolanda de Vries Mathias Hoehn 《Journal of visualized experiments : JoVE》2013,(81)
In vivo19F MRI allows quantitative cell tracking without the use of ionizing radiation. It is a noninvasive technique that can be applied to humans. Here, we describe a general protocol for cell labeling, imaging, and image processing. The technique is applicable to various cell types and animal models, although here we focus on a typical mouse model for tracking murine immune cells. The most important issues for cell labeling are described, as these are relevant to all models. Similarly, key imaging parameters are listed, although the details will vary depending on the MRI system and the individual setup. Finally, we include an image processing protocol for quantification. Variations for this, and other parts of the protocol, are assessed in the Discussion section. Based on the detailed procedure described here, the user will need to adapt the protocol for each specific cell type, cell label, animal model, and imaging setup. Note that the protocol can also be adapted for human use, as long as clinical restrictions are met. 相似文献
60.
Metabolic shifts toward glutamine regulate tumor growth,invasion and bioenergetics in ovarian cancer
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Lifeng Yang Tyler Moss Lingegowda S Mangala Juan Marini Hongyun Zhao Stephen Wahlig Guillermo Armaiz‐Pena Dahai Jiang Abhinav Achreja Julia Win Rajesha Roopaimoole Cristian Rodriguez‐Aguayo Imelda Mercado‐Uribe Gabriel Lopez‐Berestein Jinsong Liu Takashi Tsukamoto Anil K. Sood Prahlad T Ram Deepak Nagrath 《Molecular systems biology》2014,10(5)
Glutamine can play a critical role in cellular growth in multiple cancers. Glutamine‐addicted cancer cells are dependent on glutamine for viability, and their metabolism is reprogrammed for glutamine utilization through the tricarboxylic acid (TCA) cycle. Here, we have uncovered a missing link between cancer invasiveness and glutamine dependence. Using isotope tracer and bioenergetic analysis, we found that low‐invasive ovarian cancer (OVCA) cells are glutamine independent, whereas high‐invasive OVCA cells are markedly glutamine dependent. Consistent with our findings, OVCA patients’ microarray data suggest that glutaminolysis correlates with poor survival. Notably, the ratio of gene expression associated with glutamine anabolism versus catabolism has emerged as a novel biomarker for patient prognosis. Significantly, we found that glutamine regulates the activation of STAT3, a mediator of signaling pathways which regulates cancer hallmarks in invasive OVCA cells. Our findings suggest that a combined approach of targeting high‐invasive OVCA cells by blocking glutamine's entry into the TCA cycle, along with targeting low‐invasive OVCA cells by inhibiting glutamine synthesis and STAT3 may lead to potential therapeutic approaches for treating OVCAs. 相似文献