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Cell polarization in response to chemical gradients is important in development and homeostasis across eukaryota. Chemosensing cells orient toward or away from gradient sources by polarizing along a front-rear axis. Using the mating response of budding yeast as a model of chemotropic cell polarization, we found that Dcv1, a member of the claudin superfamily, influences front-rear polarity. Although Dcv1 localized uniformly on the plasma membrane (PM) of vegetative cells, it was confined to the rear of cells responding to pheromone, away from the pheromone receptor. dcv1Δ conferred mislocalization of sensory, polarity and trafficking proteins, as well as PM lipids. These phenotypes correlated with defects in pheromone-gradient tracking and cell fusion. We propose that Dcv1 helps demarcate the mating-specific front domain primarily by restricting PM lipid distribution., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2023. Published by The Company of Biologists Ltd.)

Adoptive T cell therapies (ACT) have been curative for a limited number of cancer patients. The sensitization of cancer cells to T cell killing may expand the benefit of these therapies for more patients. To this end, we use a three-step approach to identify cancer genes that disfavor T cell immunity. First, we profile gene transcripts upregulated by cancer under selection pressure from T cell killing. Second, we identify potential tumor gene targets and pathways that disfavor T cell killing using signaling pathway activation libraries and genome-wide loss-of-function CRISPR-Cas9 screens. Finally, we implement pharmacological perturbation screens to validate these targets and identify BIRC2, ITGAV, DNPEP, BCL2, and ERRα as potential ACT-drug combination candidates. Here, we establish that BIRC2 limits antigen presentation and T cell recognition of tumor cells by suppressing IRF1 activity and provide evidence that BIRC2 inhibition in combination with ACT is an effective strategy to increase efficacy., Competing Interests: Declaration of interests R.J.K., S.K.V., Y.P., and N.P.R. hold equity in and are currently employed by Lyell Immunopharma, South San Francisco, CA, USA. S.J.P. has previously held positions at Boehringer Ingelheim and NextCure Inc. and currently holds equity in NextCure Inc. N.E.S. is a scientific advisor for Vertex and Qiagen., (Published by Elsevier Inc.)

Background: Oral disintegrating films (ODFs) are one of the forms of drug delivery system with better patient compliance. The advantage is that it disintegrates quickly when placed on the tongue and has better bioavailability., Aim: The aim of this study is to develop an ODF using Vaccinium oxycoccos and Plectranthus amboinicus targeting Streptococcus mutans ., Setting and Design: This in vitro study was conducted at an institutional laboratory., Materials and Methods: The chemical composition of aqueous extracts of Vaccinium oxycoccos and Plectranthus amboinicus was examined using gas chromatography (GC)-mass spectrometry (MS). Extracts were added at its minimal inhibitory concentration (MIC) into the hydroxy propylmethyl cellulose (HPMC) polymer matrix solution to develop active ODF. The study concentrated on assessing the physical properties such as thickness of film, PH of the film, folding endurance, swelling test, disintegration, and dissolution test. Color analysis, Fourier transform infrared (FTIR), spectroscopy, and scanning electron microscope (SEM) were the mechanical properties of the film assessed., Statistical Analysis: Data were analyzed statistically, one-way analysis of variance followed by post hoc analysis for the assessment of MIC. Descriptive statistics were performed for the analysis of film properties., Results: MIC was 25 μg/ml for Vaccinium oxycoccos and 50 μg/ml for Plectranthus amboinicus . Three percentage HPMC with 1% citric acid and 1% aspartame was used to develop a polymer matrix. Films pH was between 6 and 7. FTIR and SEM analysis were done to confirm the attachment of active compound in a polymer matrix., Conclusion: Vaccinium oxycoccos and Plectranthus amboinicus showed good antibacterial activity, therefore could be a potent source to minimize the incidence of S. mutans ., Competing Interests: There are no conflicts of interest., (Copyright: © 2022 Journal of Conservative Dentistry.)

Corona virus infection (COVID-19) is increasing exponentially globally. It is also affecting pregnant women among others. Complications arising during pregnancy because of COVID-19 must be considered a health issue. The objective of the study was to analyse symptoms of pregnant women affected with COVID-19 based on the available literature. The articles were searched from Medline/PubMed, Scopus which were published till June 2020 and reviewed for the determined outcomes. The review demonstrated that common symptoms were fever, cough, nausea and myalgia. Vertical transmission of Corona virus infection was not found in any of the articles reviewed. Multicenter studies are important to better understand the pathogenesis and treatment planning for COVID-19 affected pregnant women., Competing Interests: There are no conflicts of interest., (Copyright: © 2021 Journal of Family Medicine and Primary Care.)

T cells are central to all currently effective cancer immunotherapies, but the characteristics defining therapeutically effective anti-tumor T cells have not been comprehensively elucidated. Here, we delineate four phenotypic qualities of effective anti-tumor T cells: cell expansion, differentiation, oxidative stress, and genomic stress. Using a CRISPR-Cas9-based genetic screen of primary T cells we measured the multi-phenotypic impact of disrupting 25 T cell receptor-driven kinases. We identified p38 kinase as a central regulator of all four phenotypes and uncovered transcriptional and antioxidant pathways regulated by p38 in T cells. Pharmacological inhibition of p38 improved the efficacy of mouse anti-tumor T cells and enhanced the functionalities of human tumor-reactive and gene-engineered T cells, paving the way for clinically relevant interventions., Competing Interests: Declaration of Interests The work presented here was conducted entirely at NIH. S.K.V., R.J.K., and N.P.R. are employees of Lyell Immunopharma, and S.J.P. is an employee of NextCure Inc. A patent application has been filed based on some of the findings presented in this work. The authors declare no competing interests., (Published by Elsevier Inc.)

Host conditioning has emerged as an important component of effective adoptive cell transfer-based immunotherapy for cancer. High levels of IL-1β are induced by host conditioning, but its impact on the antitumor function of T cells remains unclear. We found that the administration of IL-1β increased the population size and functionality of adoptively transferred T cells within the tumor. Most importantly, IL-1β enhanced the ability of tumor-specific T cells to trigger the regression of large, established B16 melanoma tumors in mice. Mechanistically, we showed that the increase in T cell numbers was associated with superior tissue homing and survival abilities and was largely mediated by IL-1β-stimulated host cells. In addition, IL-1β enhanced T cell functionality indirectly via its actions on radio-resistant host cells in an IL-2- and IL-15-dependent manner. Our findings not only underscore the potential of provoking inflammation to enhance antitumor immunity but also uncover novel host regulations of T cell responses., (This is a work of the U.S. Government and is not subject to copyright protection in the United States. Foreign copyrights may apply.)

A paradox of tumor immunology is that tumor-infiltrating lymphocytes are dysfunctional in situ, yet are capable of stem cell-like behavior including self-renewal, expansion, and multipotency, resulting in the eradication of large metastatic tumors. We find that the overabundance of potassium in the tumor microenvironment underlies this dichotomy, triggering suppression of T cell effector function while preserving stemness. High levels of extracellular potassium constrain T cell effector programs by limiting nutrient uptake, thereby inducing autophagy and reduction of histone acetylation at effector and exhaustion loci, which in turn produces CD8 + T cells with improved in vivo persistence, multipotency, and tumor clearance. This mechanistic knowledge advances our understanding of T cell dysfunction and may lead to novel approaches that enable the development of enhanced T cell strategies for cancer immunotherapy., (Copyright © 2019 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.)

La₂CuO₄ (LCO) perovskite nanoparticle was synthesized by microwave assisted combustion method and characterized using X-ray diffraction (XRD), diffuse reflectance spectroscopy (DRS), Fourier transformed infrared spectroscopy (FT-IR), high resolution scanning electron microscopy (HR-SEM) and energy dispersive X-ray analysis (EDX), respectively. The XRD confirmed the formation of a single orthorhombic phase. The average crystallite size of LCO samples were 39.6 nm and 46.4 nm. This LCO perovskite nanoparticle was evaluated for the conversion of methanol to formaldehyde in liquid phase batch reactor at atmospheric conditions. The conversions were studied by using different solvents like dimethyl sulfoxide (DMSO), acetonitrile, n -hexane, and temperature varying from 60 °C to 90 °C and reaction time from 1 to 4 h duration. 50 mg of catalyst, 50 mmol of DMSO, 50 mmol of methanol, temperature at 70 °C and 4 h duration was the optimized condition to achieve 100% methanol conversion and 98% selectivity of formaldehyde. The reaction pathway of methanol to formaldehyde conversion is discussed.

The main aim of this work is to develop a transparent bioactive cellulose acetate (CA) film for food packaging applications. The aim of the study is also to optimize the production methods for development of bulk and surface immobilized bioactive CA films. The vacuum drying method was optimized for the production of transparent CA films. The direct infusion and UV-assisted surface immobilization methods were investigated for the development of bioactive CA films. Reduction in the crystalline nature of CA was observed to be responsible for the production of transparent CA films. Thymol, with high antioxidant and antimicrobial properties, was examined to be the major active compound (>40%) present in the polar fraction of oregano. Retention of active compounds analyzed by High Performance Liquid Chromatography (HPLC), surface morphology analyzed by Atomic Force Microscopy (AFM), and surface chemistry analyzed by X-ray Photoelectron Spectroscopy (XPS) proved the efficiency of UV-assisted surface immobilization method. Acetyl cellulose films treated with UV irradiation at 312 nm were examined to have higher retention of active compounds. The bioactive CA films produced by bulk and surface immobilization methods showed >90% and ∼65% thymol retention, respectively. The UV-assisted surface immobilization method was found to decrease the mechanical and barrier properties of CA film. The bioactive CA films produced by bulk and surface immobilization methods were found to have retained the antioxidant and antimicrobial properties of the thymol., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The principal aim of the present study is to develop a method for the production of cellulose nanofibers, from the banana peel (BP) and bract (BB). It is also the aim of this study to produce cellulose-based biopolymers through acetyl and lauroyl modifications. The microwave digestion method and ball milling assisted ultra-sonication method was optimized for sustainable extraction of micro and nano cellulose fibers, respectively. The microwave digestion method was found to be effective in the removal of hemicellulose and lignin. Micro and nano cellulose fibers of BP and BB were found to contain type I cellulose structure. Thermal stability and crystallinity index of cellulose nanofibers were examined to be higher than it's native micro cellulose. Nano cellulose fibers were examined to be a potential source for production of acetyl and lauroyl cellulose, with a high degree of substitution and thermal stability. Hence, microwave digestion and ball milling assisted ultra-sonication method was proven to be effective in the extraction of nano cellulose fiber for development of cellulose-based polymers., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The main aim of this study is to examine the different film preparation methods (Hot air oven method, Microwave oven method, and UV irradiation method) for development of Bio-thermoplastic films from agro waste polysaccharides. The rheological properties were also studied for different combinations of tamarind seed starch (T), okra mucilage polysaccharide (OK), Residual rice bran starch (R) and sugarcane bagasse cellulose (C). Increase in concentration of okra mucilage polysaccharide and sugarcane bagasse cellulose found to highly influence the rheological viscous properties of tamarind starch solution. Increase in concentration of okra mucilage (above 1.5%) in tamarind film matrix solution significantly increased the elongation properties of starch films. Increase in concentration of cellulose (above 1%) in tamarind film matrix solution significantly increased the tensile strength of starch films. The microwave oven method was found to be the quickest method (~10 to 20 min) for biopolymer film preparation. SEM and AFM analysis revealed that UV - irradiation method produces stable bio-thermoplastic films with low surface roughness and high barrier properties. The interaction of starch molecules with other natural polysaccharide produces stable thermoplastic biopolymer films, that can be developed under different processing conditions to eradicate environmental pollution caused by petrochemical plastics., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Competing Interests: There are no conflicts of interest.

The aim of the present study is to extract potential thermoplastic polysaccharides from agricultural industrial wastes. Polysaccharides were extracted from renewable agro industrial wastes such as tamarind seeds [rich in starch (TSS)], okra head waste [rich in mucilage polysaccharide (OMP)], sugarcane bagasse [rich in cellulose (SBC)] and residual rice mill wastes [rich in starch and fiber (RS)]. Urigam variety of tamarind seed starch found to be an amylose rich starch. Different polysaccharides extracted from agro wastes were found to be having high thermal stability, except okra polysaccharide (comparatively low). X-ray diffraction pattern of tamarind seed starch proved its high crystallinity index. Crystallinity index of investigated polysaccharides were found to be in the order of SBC > TSS > RS > OMP. Chemical nature of extracted polysaccharides was confirmed by Fourier transform infrared spectroscopic analysis. Residual rice bran starch granules and tamarind seed starch globules were found to be having comparatively reduced particle size than sugarcane bagasse cellulose and okra mucilage. Scanning electron microscopic analysis revealed the cluster formations of RS granules and TSS globules. Residual rice bran starch found to be associated with other fibers (present in outer coat of rice). Okra mucilage and SBC were examined to be having linear sheets and linear bundles structures, respectively., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The main aim of the present study is to extract and characterize cashew nut shell (CNS) starch and walnut shell cellulose (WNC) for development of cellulose reinforced starch films. Moreover, the extraction and characterization of pomegranate peel extract, for incorporation with CNS-WNC films, was investigated. CNS starch was examined to be a moderate amylose starch with 26.32 ± 0.43% amylose content. Thermal degradation temperature of CNS starch was found to be 310 °C. Walnut shell cellulose was found to have high crystallinity index of 72%, with two thermal degradation temperatures of 319 °C and 461 °C. 2% WN cellulose reinforced CNS starch films were examined to have good oxygen transfer rate, mechanical and physical properties. Thermal degradation temperature of CNS-WNC starch films were found to be at the range of 298-302 °C. Surface roughness of CNS-WNC starch films were found to be increasing with increase in concentration of cellulose in films. Hydroxymethylfurfurole, Benzene, 2-methoxy-1,3,4-trimethyl and 1,2,3-Propanetriol, 1-acetate were found to be major active compounds present in hydrophilic extracts of Punica granatum peels. 2% WN cellulose reinforced starch films infused with hydrophilic active compounds of pomegranate peel was examined to be having good active package properties., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Adoptive immunotherapies using T cells genetically redirected with a chimeric antigen receptor (CAR) or T cell receptor (TCR) are entering mainstream clinical practice. Despite encouraging results, some patients do not respond to current therapies. In part, this phenomenon has been associated with infusion of reduced numbers of early memory T cells. Herein, we report that AKT signaling inhibition is compatible with CAR and TCR retroviral transduction of human T cells while promoting a CD62L-expressing central memory phenotype. Critically, this intervention did not compromise cell yield. Mechanistically, disruption of AKT signaling preserved MAPK activation and promoted the intranuclear localization of FOXO1, a transcriptional regulator of T cell memory. Consequently, AKT signaling inhibition synchronized the transcriptional profile for FOXO1-dependent target genes across multiple donors. Expression of an AKT-resistant FOXO1 mutant phenocopied the influence of AKT signaling inhibition, while addition of AKT signaling inhibition to T cells expressing mutant FOXO1 failed to further augment the frequency of CD62L-expressing cells. Finally, treatment of established B cell acute lymphoblastic leukemia was superior using anti-CD19 CAR-modified T cells transduced and expanded in the presence of an AKT inhibitor compared with conventionally grown T cells. Thus, inhibition of signaling along the PI3K/AKT axis represents a generalizable strategy to generate large numbers of receptor-modified T cells with an early memory phenotype and superior antitumor efficacy.

Brain-derived neurotrophic factor (BDNF) is an activity-dependent neurotrophin critical for neuronal plasticity in the hippocampus. BDNF is encoded by multiple transcripts with alternative 5' untranslated regions (5'UTRS) that display activity-induced targeting to distinct subcellular compartments. While individual Bdnf 5'UTR transcripts influence dendrite morphology in cultured hippocampal neurons, it is unknown whether Bdnf splice variants impact dendrite arborization in functional classes of neurons in the intact hippocampus. Moreover, the contribution of Bdnf 5'UTR splice variants to dendritic spine density and shape has not been explored. We analyzed the structure of CA1 and CA3 dendrite arbors in transgenic mice lacking BDNF production from exon (Ex) 1, 2, 4, or 6 splice variants (Bdnf-e1, -e2, -e4, and -e6 -/- mice) and found that loss of BDNF from individual Bdnf mRNA variants differentially impacts the complexity of apical and basal arbors in vivo. Consistent with the subcellular localization studies, Bdnf Ex2 and Ex6 transcripts significantly contributed to dendrite morphology in both CA1 and CA3 neurons. While Bdnf-e2 -/- mice showed increased branching proximal to the soma in CA1 and CA3 apical arbors, Bdnf-e6 -/- mice showed decreased apical and basal dendrite complexity. Analysis of spine morphology on Bdnf-e6 -/- CA1 dendrites revealed changes in the percentage of differently sized spines on apical, but not basal, branches. These results provide further evidence that Bdnf splice variants generate a spatial code that mediates the local actions of BDNF in distinct dendritic compartments on structural and functional plasticity.

Somatic gene mutations can alter the vulnerability of cancer cells to T-cell-based immunotherapies. Here we perturbed genes in human melanoma cells to mimic loss-of-function mutations involved in resistance to these therapies, by using a genome-scale CRISPR-Cas9 library that consisted of around 123,000 single-guide RNAs, and profiled genes whose loss in tumour cells impaired the effector function of CD8 + T cells. The genes that were most enriched in the screen have key roles in antigen presentation and interferon-γ signalling, and correlate with cytolytic activity in patient tumours from The Cancer Genome Atlas. Among the genes validated using different cancer cell lines and antigens, we identified multiple loss-of-function mutations in APLNR, encoding the apelin receptor, in patient tumours that were refractory to immunotherapy. We show that APLNR interacts with JAK1, modulating interferon-γ responses in tumours, and that its functional loss reduces the efficacy of adoptive cell transfer and checkpoint blockade immunotherapies in mouse models. Our results link the loss of essential genes for the effector function of CD8 + T cells with the resistance or non-responsiveness of cancer to immunotherapies.

This study investigates the integrated approach of spice extracts and modified atmospheric packaging (MAP) chicken meat preservation. Specifically, extracts from clove (CL), cinnamon (CI) individually and in combination (3% w/w) along with MAP (30% CO2/70% N2 and 10% O2/30% CO2/60% N2) were used to increase the shelf life of fresh chicken meat stored at 4°C. The parameters evaluated as shelf life indications are microbiological (total viable count, Pseudomonas spp., lactic acid bacteria (LAB), and Enterobacteriaceae), physicochemical (pH, Lipid oxidation, color changes) and Sensory attributes. Microbial population were reduced by 2.5 to 5 log cfu/g, with the greater impact being accomplished by the blend of clove and cinnamon extract with 30% CO2/70% N2 MAP. Thiobarbituric values for all treated and MAP packed samples remained lower than 1 mg malondialdehyde (MDA)/kg all through the 24 day storage period. pH values varied from 5.5 for fresh sample on day 0 to 7.11 (day 25) on combined extract treated and MAP packaged samples. The estimations of the color parameters L*, a*, and b* were well maintained in oxygen deficient MAP. Finally, sensory investigation demonstrated that combined clove and cinnamon extract of 3% conferred acceptable sensory attributes to the samples on day 24 of storage. These results indicate the extended shelf life of chicken meat from 4 days to 24 days for samples when coated with 3% of combined clove and cinnamon extract and packaged under MAP without oxygen. These pooled extracts along with MAP displayed expanded the usability and the organoleptic qualities of chicken meat., (© 2017 Poultry Science Association Inc.)

Cancer immunotherapy is an increasingly successful strategy for the treatment of patients who have advanced or conventional therapy-resistant cancers. T cells are key mediators of tumor destruction and their specificity for tumor-expressed antigens is of paramount importance, but other T cell-intrinsic qualities, such as durability, longevity, and functionality also play important roles in determining the efficacy of immunotherapy. The cellular energetic pathways that are utilized by T cells play a key role in regulating each of these qualities. Metabolic activity, which both regulates and is regulated by cellular signaling pathways and epigenetics, also profoundly influences the trajectories of T cell differentiation and fate. In this Review, we discuss how cell metabolism influences T cell anti-tumor activity, the metabolic qualities of highly-functional T cells, and strategies to modulate metabolism for improving the immune response to tumors., (Published by Elsevier Inc.)

Competing Interests: There are no conflicts of interest.

In the present study, mathematical models were used to examine the effect of active compound diffusion from edible film (supplemented with S. aromaticum and C. cassia) on the microbial, physical and chemical quality of mutton stored at 4 and 10°C. Cinnamaldehyde and eugenol release from edible film into liquid was found to be 80% and 75% of the equilibrium concentration. Active compound release into meat was 42-51% for cinnamaldehyde and 38-48% for eugenol, in storage temperatures of 4-15°C. Developed mathematical models showed the diffusivity of cinnamaldehyde (0.45×10 -15 ±0.04×10 -15 ) and eugenol (0.63×10 -10 ±0.01×10 -10 ) into meat, which was about 40% of that in liquid medium. On comparing physical, chemical and microbial results, shelf life of mutton meat was found to be increased by 1week at storage temperature of 10°C and 3weeks at storage temperature of 4°C., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Immunotherapies designed to trigger T cell destruction of tumor cells can result in sustained and complete responses in patients whose cancers were resistant to available treatment options. Evidence suggests that powering the T cell response - how T cells generate energy - plays an important role in their effectiveness. Furthermore the metabolism of T cells can be modulated to improve their anti-cancer activities. In this review, we will discuss the key metabolic properties of anti-cancer T cells, along with potential strategies to enhance immunotherapy through the modulation of T cell metabolism., (Published by Elsevier Ltd.)

A clear role for cellular metabolism in regulating immune cell function has recently been established. In a recent issue of Cell, Geiger et al. (2016) demonstrate a role for intracellular arginine abundance in T cells for the promotion of oxidative metabolism, increased cell viability, persistence, and in vivo antitumor response., (Published by Elsevier Inc.)

Tumours progress despite being infiltrated by tumour-specific effector T cells. Tumours contain areas of cellular necrosis, which are associated with poor survival in a variety of cancers. Here, we show that necrosis releases intracellular potassium ions into the extracellular fluid of mouse and human tumours, causing profound suppression of T cell effector function. Elevation of the extracellular potassium concentration ([K + ] e ) impairs T cell receptor (TCR)-driven Akt-mTOR phosphorylation and effector programmes. Potassium-mediated suppression of Akt-mTOR signalling and T cell function is dependent upon the activity of the serine/threonine phosphatase PP2A. Although the suppressive effect mediated by elevated [K + ] e is independent of changes in plasma membrane potential (V m ), it requires an increase in intracellular potassium ([K + ] i ). Accordingly, augmenting potassium efflux in tumour-specific T cells by overexpressing the potassium channel K v 1.3 lowers [K + ] i and improves effector functions in vitro and in vivo and enhances tumour clearance and survival in melanoma-bearing mice. These results uncover an ionic checkpoint that blocks T cell function in tumours and identify potential new strategies for cancer immunotherapy., Competing Interests: The authors declare no competing financial interests.

Cancer cells must evade immune responses at distant sites to establish metastases. The lung is a frequent site for metastasis. We hypothesized that lung-specific immunoregulatory mechanisms create an immunologically permissive environment for tumor colonization. We found that T-cell-intrinsic expression of the oxygen-sensing prolyl-hydroxylase (PHD) proteins is required to maintain local tolerance against innocuous antigens in the lung but powerfully licenses colonization by circulating tumor cells. PHD proteins limit pulmonary type helper (Th)-1 responses, promote CD4(+)-regulatory T (Treg) cell induction, and restrain CD8(+) T cell effector function. Tumor colonization is accompanied by PHD-protein-dependent induction of pulmonary Treg cells and suppression of IFN-γ-dependent tumor clearance. T-cell-intrinsic deletion or pharmacological inhibition of PHD proteins limits tumor colonization of the lung and improves the efficacy of adoptive cell transfer immunotherapy. Collectively, PHD proteins function in T cells to coordinate distinct immunoregulatory programs within the lung that are permissive to cancer metastasis. PAPERCLIP., (Published by Elsevier Inc.)

T cell antigen receptor (TCR) signaling drives distinct responses depending on the differentiation state and context of CD8(+) T cells. We hypothesized that access of signal-dependent transcription factors (TFs) to enhancers is dynamically regulated to shape transcriptional responses to TCR signaling. We found that the TF BACH2 restrains terminal differentiation to enable generation of long-lived memory cells and protective immunity after viral infection. BACH2 was recruited to enhancers, where it limited expression of TCR-driven genes by attenuating the availability of activator protein-1 (AP-1) sites to Jun family signal-dependent TFs. In naive cells, this prevented TCR-driven induction of genes associated with terminal differentiation. Upon effector differentiation, reduced expression of BACH2 and its phosphorylation enabled unrestrained induction of TCR-driven effector programs., Competing Interests: The authors declare no competing financial interests.

To better elucidate epigenetic mechanisms that correlate with the dynamic gene expression program observed upon T-cell differentiation, we investigated the genomic landscape of histone modifications in naive and memory CD8(+) T cells. Using a ChIP-Seq approach coupled with global gene expression profiling, we generated genome-wide histone H3 lysine 4 (H3K4me3) and H3 lysine 27 (H3K27me3) trimethylation maps in naive, T memory stem cells, central memory cells, and effector memory cells in order to gain insight into how histone architecture is remodeled during T cell differentiation. We show that H3K4me3 histone modifications are associated with activation of genes, while H3K27me3 is negatively correlated with gene expression at canonical loci and enhancers associated with T-cell metabolism, effector function, and memory. Our results also reveal histone modifications and gene expression signatures that distinguish the recently identified T memory stem cells from other CD8(+) T-cell subsets. Taken together, our results suggest that CD8(+) lymphocytes undergo chromatin remodeling in a progressive fashion. These findings have major implications for our understanding of peripheral T-cell ontogeny and the formation of immunological memory.

Brain-derived neurotrophic factor (BDNF) regulates diverse biological functions ranging from neuronal survival and differentiation during development to synaptic plasticity and cognitive behavior in the adult. BDNF disruption in both rodents and humans is associated with neurobehavioral alterations and psychiatric disorders. A unique feature of Bdnf transcription is regulation by nine individual promoters, which drive expression of variants that encode an identical protein. It is hypothesized that this unique genomic structure may provide flexibility that allows different factors to regulate BDNF signaling in distinct cell types and circuits. This has led to the suggestion that isoforms may regulate specific BDNF-dependent functions; however, little scientific support for this idea exists. We generated four novel mutant mouse lines in which BDNF production from one of the four major promoters (I, II, IV, or VI) is selectively disrupted (Bdnf-e1, -e2, -e4, and -e6 mice) and used a comprehensive comparator approach to determine whether different Bdnf transcripts are associated with specific BDNF-dependent molecular, cellular, and behavioral phenotypes. Bdnf-e1 and -e2 mutant males displayed heightened aggression accompanied by convergent expression changes in specific genes associated with serotonin signaling. In contrast, BDNF-e4 and -e6 mutants were not aggressive but displayed impairments associated with GABAergic gene expression. Moreover, quantifications of BDNF protein in the hypothalamus, prefrontal cortex, and hippocampus revealed that individual Bdnf transcripts make differential, region-specific contributions to total BDNF levels. The results highlight the biological significance of alternative Bdnf transcripts and provide evidence that individual isoforms serve distinct molecular and behavioral functions.

The immune system has a powerful ability to recognize and kill cancer cells, but its function is often suppressed within tumors, preventing clearance of disease. Functionally diverse innate and adaptive cellular lineages either drive or constrain immune reactions within tumors. The transcription factor (TF) BACH2 regulates the differentiation of multiple innate and adaptive cellular lineages, but its role in controlling tumor immunity has not been elucidated. Here, we demonstrate that BACH2 is required to establish immunosuppression within tumors. Tumor growth was markedly impaired in Bach2-deficient mice and coincided with intratumoral activation of both innate and adaptive immunity. However, augmented tumor clearance in the absence of Bach2 was dependent upon the adaptive immune system. Analysis of tumor-infiltrating lymphocytes from Bach2-deficient mice revealed high frequencies of rapidly proliferating effector CD4+ and CD8+ T cells that expressed the inflammatory cytokine IFN-γ. Effector T cell activation coincided with a reduction in the frequency of intratumoral Foxp3+ Tregs. Mechanistically, BACH2 promoted tumor immunosuppression through Treg-mediated inhibition of intratumoral CD8+ T cells and IFN-γ. These findings demonstrate that BACH2 is a key component of the molecular program of tumor immunosuppression and identify therapeutic targets for the reversal of immunosuppression in cancer.

Starch from Tamarind seed is considered to be a nonedible and inexpensive component, with many industrial applications. Extraction and characterization of tamarind seed starch was carried out for the synthesis of biopolymer. Tamarind seeds were collected, cleaned and further roasted, decorticated, and pulverized to get starch powder. Total starch content present in each tamarind seed is estimated to be around 65-70%. About 84.68% purified starch can be recovered from the tamarind seed. Defatted Tamarind seed starch has an amylose content of 27.55 wt.% and 72.45 wt.% of amylopectin. Morphological (SEM) and X-ray diffraction were used to evaluate crystallinity. Likewise, TGA and DSC of starch have also been analyzed. Thermal properties of starch obtained from tamarind seeds showed good thermal stability when compared to other starch sources such as Mesquite seed and Mango kernel. This study proved that the tamarind seed starch can be used as a potential biopolymer material. Thermo-stable biofilms were produced through initial optimization studies. Predictive response surface quadratic models were constructed for prediction and optimization of biofilm mechanical properties. Correlation coefficient values were calculated to me more than 0.90 for mechanical responses which implies the fitness of constructed model with experimental data., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Long-term survival and antitumor immunity of adoptively transferred CD8(+) T cells is dependent on their metabolic fitness, but approaches to isolate therapeutic T cells based on metabolic features are not well established. Here we utilized a lipophilic cationic dye tetramethylrhodamine methyl ester (TMRM) to identify and isolate metabolically robust T cells based on their mitochondrial membrane potential (ΔΨm). Comprehensive metabolomic and gene expression profiling demonstrated global features of improved metabolic fitness in low-ΔΨm-sorted CD8(+) T cells. Transfer of these low-ΔΨm T cells was associated with superior long-term in vivo persistence and an enhanced capacity to eradicate established tumors compared with high-ΔΨm cells. Use of ΔΨm-based sorting to enrich for cells with superior metabolic features was observed in CD8(+), CD4(+) T cell subsets, and long-term hematopoietic stem cells. This metabolism-based approach to cell selection may be broadly applicable to therapies involving the transfer of HSC or lymphocytes for the treatment of viral-associated illnesses and cancer., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Adoptive cell transfer (ACT) of purified naive, stem cell memory, and central memory T cell subsets results in superior persistence and antitumor immunity compared with ACT of populations containing more-differentiated effector memory and effector T cells. Despite a clear advantage of the less-differentiated populations, the majority of ACT trials utilize unfractionated T cell subsets. Here, we have challenged the notion that the mere presence of less-differentiated T cells in starting populations used to generate therapeutic T cells is sufficient to convey their desirable attributes. Using both mouse and human cells, we identified a T cell-T cell interaction whereby antigen-experienced subsets directly promote the phenotypic, functional, and metabolic differentiation of naive T cells. This process led to the loss of less-differentiated T cell subsets and resulted in impaired cellular persistence and tumor regression in mouse models following ACT. The T memory-induced conversion of naive T cells was mediated by a nonapoptotic Fas signal, resulting in Akt-driven cellular differentiation. Thus, induction of Fas signaling enhanced T cell differentiation and impaired antitumor immunity, while Fas signaling blockade preserved the antitumor efficacy of naive cells within mixed populations. These findings reveal that T cell subsets can synchronize their differentiation state in a process similar to quorum sensing in unicellular organisms and suggest that disruption of this quorum-like behavior among T cells has potential to enhance T cell-based immunotherapies.

Pharmacologic inhibitors of the serine/threonine kinase Akt, initially aimed at deranged oncogenic pathways in tumors, have recently been shown to act as immunomodulators that markedly enhance the antitumor properties of T cells. Repurposing Akt inhibitors to improve antitumor immunity may be viewed as a manifestation of a larger paradigmatic shift in which hallmark characteristics of cancer (e.g., immune evasion), rather than merely causal features (e.g., somatic mutations) can be exploited for therapeutic benefit.

Aim: To investigate the effect of corn starch (CS) edible films with spice oils on the stability of raw beef during refrigerated storage., Methods and Results: The antimicrobial properties of corn starch films containing 0-4·0% (w/v) ratios of clove (CL) and cinnamon (CI) essential oils (EOs) were tested against seven meat spoilage organisms by zone inhibition test. Effects of CS films containing 3% CL or CI or a mixture of CL + CI were also tested in raw beef stored at 4°C. Meat samples were analysed for pH, microbial counts, colour values and Thiobarbituric acid reactive substances (TBARS) values for a period of 15 days. CS films with CL + CI resulted in a significant reduction in microbial populations in the meat samples and also improved meat colour stability at the end of storage period., Conclusions: The incorporation of spice EOs in CS films may provide antimicrobial and antioxidant activities that could improve the stability of raw meat., Significance and Impact of the Study: Results from this study may be applied in meat industries as an additional barrier to control microbial spoilage as well as lipid oxidation in meat products., (© 2015 The Society for Applied Microbiology.)

Improving the functional avidity of effector T cells is critical in overcoming inhibitory factors within the tumor microenvironment and eliciting tumor regression. We have found that Cish, a member of the suppressor of cytokine signaling (SOCS) family, is induced by TCR stimulation in CD8(+) T cells and inhibits their functional avidity against tumors. Genetic deletion of Cish in CD8(+) T cells enhances their expansion, functional avidity, and cytokine polyfunctionality, resulting in pronounced and durable regression of established tumors. Although Cish is commonly thought to block STAT5 activation, we found that the primary molecular basis of Cish suppression is through inhibition of TCR signaling. Cish physically interacts with the TCR intermediate PLC-γ1, targeting it for proteasomal degradation after TCR stimulation. These findings establish a novel targetable interaction that regulates the functional avidity of tumor-specific CD8(+) T cells and can be manipulated to improve adoptive cancer immunotherapy.

It is thought that cancer cells engage in Warburg metabolism to meet intrinsic biosynthetic requirements of cell growth and proliferation. Papers by Chang et al. and Ho et al. show that Warburg metabolism enables tumor cells to restrict glucose availability to T cells, suppressing anti-tumor immunity., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Background: Neurogenesis continues throughout life in the hippocampal dentate gyrus. Chronic treatment with monoaminergic antidepressant drugs stimulates hippocampal neurogenesis, and new neurons are required for some antidepressant-like behaviors. Electroconvulsive seizures (ECS), a laboratory model of electroconvulsive therapy (ECT), robustly stimulate hippocampal neurogenesis., Hypothesis: ECS requires newborn neurons to improve behavioral deficits in a mouse neuroendocrine model of depression., Methods: We utilized immunohistochemistry for doublecortin (DCX), a marker of migrating neuroblasts, to assess the impact of Sham or ECS treatments (1 treatment per day, 7 treatments over 15 days) on hippocampal neurogenesis in animals receiving 6 weeks of either vehicle or chronic corticosterone (CORT) treatment in the drinking water. We conducted tests of anxiety- and depressive-like behavior to investigate the ability of ECS to reverse CORT-induced behavioral deficits. We also determined whether adult neurons are required for the effects of ECS. For these studies we utilized a pharmacogenetic model (hGFAPtk) to conditionally ablate adult born neurons. We then evaluated behavioral indices of depression after Sham or ECS treatments in CORT-treated wild-type animals and CORT-treated animals lacking neurogenesis., Results: ECS is able to rescue CORT-induced behavioral deficits in indices of anxiety- and depressive-like behavior. ECS increases both the number and dendritic complexity of adult-born migrating neuroblasts. The ability of ECS to promote antidepressant-like behavior is blocked in mice lacking adult neurogenesis., Conclusion: ECS ameliorates a number of anxiety- and depressive-like behaviors caused by chronic exposure to CORT. ECS requires intact hippocampal neurogenesis for its efficacy in these behavioral indices., (Copyright © 2015 Elsevier Inc. All rights reserved.)

The present study evaluates the oxidative stability in chikkis enriched with omega 3 fatty acids using natural antioxidant from Rosmarinus officinalis. The best condition for the extraction of phenolic compounds from Rosmarinus officinalis L. (rosemary) was established, and the antioxidant activity was demonstrated based on inhibition of DPPH free radical formation. Nannochloropsis oculata and Isochrysis galbana are rich sources of Eicosapentanoic acid (EPA) and Docosahexaenoic acid (DHA). Biomass of these microalgae were incorporated in chikkis as omega 3 fatty acid source. Effects of addition of natural and synthetic antioxidants (BHA) on oxidative stability of chikkis were analyzed for storage period of 2 months. Evaluation of peroxide value (PV) and fatty acid profile showed that the process of oxidation slowed down. Natural antioxidant was found to be more effective when compared to synthetic antioxidant (BHA). Omega-3 PUFA levels (EPA+DHA) of 75 and 240 mg/100 g chikkis were observed if enriched with 1 and 3 % Nannochloropsis oculata biomass respectively. Similarly, Omega-3 PUFA levels (EPA+DHA) of 102 and 320 mg/100 g chikkis were observed if enriched with 1 and 3 % Isochrysis galbana biomass respectively. The effects of microalgae and antioxidant incorporation on the chikkis showed that color values remained stable during storage period of 2 months with no significant change (P < 0.05) in texture. Sensory evaluation revealed that up to 3 % microalgal biomass incorporation was positively classified and accepted.

A high-throughput method is described, consisting of a colourimetric microtitration for screening phenol-degrading microorganisms, using a mixture of 4-aminoantipyrine and potassium ferricyanide as the colour indicator. This contemporary study summarizes a new method to determine phenol-degrading bacteria isolated from different areas. The method was used for testing a total of 72 bacteria collected from the natural environment and five known strains obtained from diagnostic and research laboratories employing 200 mg/L phenol (the linear range saturation concentration). Depending on the change in colour indicator, the degradation profiles of 11 strains of bacteria are shown, of which seven strains were able to degrade more than 80 % of phenol within 6-8 h, while the other four strains took 12-24 h. Two of the environmentally isolated strains showed high efficiency of phenol degradation and were confirmed by the high-performance liquid chromatography analysis. These strains were identified by 16S rRNA sequencing as unique (Escherichia coli moh1 and Bacillus cereus moh2) and were deposited in the GenBank of NCBI. Two pathogenic strains (Uropathogenic E. coli and Salmonella sp.) were found to be the fast degraders of phenol, which is of medical concern, as phenol is generally used as a disinfectant in hospitals. This method can be used for the estimation and screening of phenol degraders in a single step, for its application in bioremediation as well as in hospitals for screening the phenol resistance of pathogens.

Adoptive cell therapy (ACT) using autologous tumor-infiltrating lymphocytes (TIL) results in complete regression of advanced cancer in some patients, but the efficacy of this potentially curative therapy may be limited by poor persistence of TIL after adoptive transfer. Pharmacologic inhibition of the serine/threonine kinase Akt has recently been shown to promote immunologic memory in virus-specific murine models, but whether this approach enhances features of memory (e.g., long-term persistence) in TIL that are characteristically exhausted and senescent is not established. Here, we show that pharmacologic inhibition of Akt enables expansion of TIL with the transcriptional, metabolic, and functional properties characteristic of memory T cells. Consequently, Akt inhibition results in enhanced persistence of TIL after adoptive transfer into an immunodeficient animal model and augments antitumor immunity of CD8 T cells in a mouse model of cell-based immunotherapy. Pharmacologic inhibition of Akt represents a novel immunometabolomic approach to enhance the persistence of antitumor T cells and improve the efficacy of cell-based immunotherapy for metastatic cancer., (©2014 American Association for Cancer Research.)

Lymphodepleting regimens are used before adoptive immunotherapy to augment the antitumor efficacy of transferred T cells by removing endogenous homeostatic "cytokine sinks." These conditioning modalities, however, are often associated with severe toxicities. We found that microRNA-155 (miR-155) enabled tumor-specific CD8(+) T cells to mediate profound antitumor responses in lymphoreplete hosts that were not potentiated by immune-ablation. miR-155 enhanced T-cell responsiveness to limited amounts of homeostatic γc cytokines, resulting in delayed cellular contraction and sustained cytokine production. miR-155 restrained the expression of the inositol 5-phosphatase Ship1, an inhibitor of the serine-threonine protein kinase Akt, and multiple negative regulators of signal transducer and activator of transcription 5 (Stat5), including suppressor of cytokine signaling 1 (Socs1) and the protein tyrosine phosphatase Ptpn2. Expression of constitutively active Stat5a recapitulated the survival advantages conferred by miR-155, whereas constitutive Akt activation promoted sustained effector functions. Our results indicate that overexpression of miR-155 in tumor-specific T cells can be used to increase the effectiveness of adoptive immunotherapies in a cell-intrinsic manner without the need for life-threatening, lymphodepleting maneuvers.

Both targeted inhibition of oncogenic driver mutations and immune-based therapies show efficacy in treatment of patients with metastatic cancer, but responses can be either short lived or incompletely effective. Oncogene inhibition can augment the efficacy of immune-based therapy, but mechanisms by which these two interventions might cooperate are incompletely resolved. Using a novel transplantable BRAF(V600E)-mutant murine melanoma model (SB-3123), we explored potential mechanisms of synergy between the selective BRAF(V600E) inhibitor vemurafenib and adoptive cell transfer (ACT)-based immunotherapy. We found that vemurafenib cooperated with ACT to delay melanoma progression without significantly affecting tumor infiltration or effector function of endogenous or adoptively transferred CD8(+) T cells, as previously observed. Instead, we found that the T-cell cytokines IFNγ and TNFα synergized with vemurafenib to induce cell-cycle arrest of tumor cells in vitro. This combinatorial effect was recapitulated in human melanoma-derived cell lines and was restricted to cancers bearing a BRAF(V600E) mutation. Molecular profiling of treated SB-3123 indicated that the provision of vemurafenib promoted the sensitization of SB-3123 to the antiproliferative effects of T-cell effector cytokines. The unexpected finding that immune cytokines synergize with oncogene inhibitors to induce growth arrest has major implications for understanding cancer biology at the intersection of oncogenic and immune signaling and provides a basis for design of combinatorial therapeutic approaches for patients with metastatic cancer., (©2014 American Association for Cancer Research.)

Microbial desalination cell (MDC) is a bioelectrochemical system developed recently from microbial fuel cells (MFCs), for producing green energy from organic wastes along with desalination of saltwater. MDC is proved to be a better performer than MFC in terms of power output and chemical oxygen demand removal, with desalination as an additional feature. This study investigates the application potential of MDC for integrated biodegradation of waste engine oil. This study showed, for the first time, that waste engine oil could be used as an organic substrate in MDC, achieving biodegradation of engine oil along with considerable desalination and power production. Utilization of these wastes in MDC can protect the environment from waste engine oil contamination. Indigenous oil-degrading bacteria were isolated and identified from engine oil contaminated sludge. Degradation of waste engine oil by these novel isolates was studied in batch cultures and optimized the growth conditions. The same cultures when used in MDC, gave enhanced biodegradation (70.1 +/- 0.5%) along with desalination (68.3 +/- 0.6%) and power production (3.1 +/- 0.3 mW/m2). Fourier transform-infrared spectroscopy and gas chromatography-mass spectrometry analyses were performed to characterize the degradation metabolites in the anolyte of MDC which clearly indicated the biodegradation of long chain, branched and cyclic hydrocarbons present in waste engine oil.

Background: This study examined the effectiveness of three individual spice (clove, cinnamon and oregano) extracts and their combinations in raw beef meat during refrigerated storage. Meat samples were monitored for microbiological (total viable count, Enterobacteriaceae, lactic acid bacteria, Brochothrix thermosphacta and Pseudomonas spp.) and physicochemical (pH, colour and thiobarbituric acid-reactive substances (TBARS)) attributes., Results: Samples treated with the combination of all three spice extracts showed lower bacterial counts and better L*, a* and b* values among treated samples during the storage period. Positive and negative control samples had the highest TBARS values at the end of the storage period. With the addition of spice extracts, TBARS values in raw beef samples were retarded effectively (P < 0.05) compared with control samples, especially when the combination of all three spice extracts was used., Conclusion: The results of this study show that spice extracts were effective in inhibiting the growth of microbial populations and retarding lipid oxidation during refrigerated storage (4 °C) of raw beef meat. They also suggest that combinations of these extracts may have potential as natural preservatives in raw meat products., (© 2014 Society of Chemical Industry.)

Biomass and lipid productivities of Isochrysis galbana were optimized using nutrients of molasses (4, 8, 12 g l(-1)), glucose (4, 8, 12 g l(-1)), glycerol (4, 8, 12 g l(-1)) and yeast extract (2 g l(-1)). Combinations of carbon sources at different ratios were evaluated in which the alga was grown at three different light intensities (50, 100 and 150 μmol m(-2) s(-1)) under the influence of three different photoperiod cycles (12/12, 18/6 and 24/0 h light/dark). A maximum cell density of 8.35 g l(-1) with 32 % (w/w) lipid was achieved for mixotrophic growth at 100 μmol m(-2) s(-1) and 18/6 h light/dark with molasses/glucose (20:80 w/w). Mixotrophic cultivation using molasses, glucose and glycerol was thus effective for the cultivation of I. galbana.

The antimicrobial and antioxidant effects of different spice extracts in raw chicken meat during storage for 15 days at 4 °C were studied. Raw chicken meat was treated with BHT (positive control), Syzygium aromaticum (SA), Cinnmomum cassia (CC), Origanum vulgare (OV), and Brassica nigra (BN) extracts and the different combinations as well as the results were compared to raw chicken meat without any additive (negative control). The antioxidant and antimicrobial activities of spice extracts were determined. Total phenolic contents and flavonoid contents were ranged from 14.09 ± 0.78 to 24.65 ± 0.83 mg of GAE/g and 7.07 ± 0.15 to 12.13 ± 0.24 mg of quercetin/g, respectively. The pH, instrumental color (CIE L*, a*, b*), total viable counts (TVC), Lactic Acid Bacteria (LAB) counts, Enterobacteriaceae counts, Pseudomonas spp. counts and 2-thiobarbituric acid reactive substances (TBARS) were determined at a gap of 3 days interval for a period of 15 days. The bacterial counts of T-W-SA+T-W-CC+T-W-OV samples were lower than control samples during storage. T-W-SA+T-W-CC+T-W-OV samples maintained significantly (P<0.05) higher L*, a* and b* values while storing. The TBARS values of T-W-SA+T-W-CC+T-W-OV samples were lowest among the samples. These results show that spice extracts are very effective against microbial growth, lipid oxidation and has potential as a natural antioxidant in raw chicken meats., (Copyright © 2013 Elsevier B.V. All rights reserved.)