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We report here that nucleus accumbens–associated protein-1 (NAC1), a nuclear factor of the Broad-complex, Tramtrack, Bric-a-brac/poxvirus and zinc finger (BTB/POZ) gene family, is a negative regulator of FoxP3 in regulatory T cells (T regs ) and a critical determinant of immune tolerance. Phenotypically, NAC1 −/− mice showed substantial tolerance to the induction of autoimmunity and generated a larger amount of CD4 + T regs that exhibit a higher metabolic profile and immune-suppressive activity, increased acetylation and expression of FoxP3, and slower turnover of this transcription factor. Treatment of T regs with the proinflammatory cytokines interleukin-1β or tumor necrosis factor–α induced a robust up-regulation of NAC1 but evident down-regulation of FoxP3 as well as the acetylated FoxP3. These findings imply that NAC1 acts as a trigger of the immune response through destabilization of T regs and suppression of tolerance induction, and targeting of NAC1 warrants further exploration as a potential tolerogenic strategy for treatment of autoimmune disorders.

Immunotherapy has led to impressive advances in the treatment of autoimmune and pro-inflammatory disorders; yet, its clinical outcomes remain limited by a variety of factors including the pro-inflammatory microenvironment (IME). Discovering effective immunomodulatory agents, and the mechanisms by which they control disease, will lead to innovative strategies for enhancing the effectiveness of current immunotherapeutic approaches. We have metabolically engineered an attenuated bacterial strain (i.e., Brucella melitensis 16M ΔvjbR, BmΔvjbR) to produce indole, a tryptophan metabolite that controls the fate and function of regulatory T cells (Tregs). We demonstrated that treatment with this strain polarized M2 macrophages (Mφ) which produced anti-inflammatory cytokines (e.g., IL-10) and promoted Treg function; moreover, when combined with adoptive cell transfer (ACT) of Tregs, a single treatment with our engineered bacterial strain dramatically reduced the incidence and score of autoimmune arthritis and decreased joint damage. These findings show how a metabolically engineered bacterium can constitute a powerful vehicle for improving the efficacy of immunotherapy, defeating autoimmunity and reducing inflammation by remodeling the IME and augmenting Treg function.

eEF-2K has important roles in stress responses and cellular metabolism. We report here a previously unappreciated but critical role of eEF-2K in regulating the fate and cytocidal activity of CD8 + T cells. CD8 + T cells from eEF-2K KO mice were more proliferative but had lower survival than their wild-type counterparts after their activation, followed by occurrence of premature senescence and exhaustion. eEF-2K KO CD8 + T cells were more metabolically active and showed hyperactivation of the Akt-mTOR-S6K pathway. Loss of eEF-2K substantially impaired the activity of CD8 + T cells. Furthermore, the antitumor efficacy and tumor infiltration of the CAR-CD8 + T cells lacking eEF-2K were notably reduced as compared to the control CAR-CD8 + T cells. Thus, eEF-2K is critically required for sustaining the viability and function of cytotoxic CD8 + T cells, and therapeutic augmentation of this kinase may be exploited as a novel approach to reinforcing CAR-T therapy against cancer.

The viral antigen (Ag)-specific CD8+ cytotoxic T lymphocytes (CTLs) derived from pluripotent stem cells (PSCs), i.e., PSC-CTLs, have the ability to suppress the human immunodeficiency virus (HIV) infection. After adoptive transfer, PSC-CTLs can infiltrate into the local tissues to suppress HIV replication. Nevertheless, the mechanisms by which the viral Ag-specific PSC-CTLs elicit the antiviral response remain to be fully elucidated. In this study, we generated the functional HIV-1 Gag epitope SL9-specific CTLs from the induced PSC (iPSCs), i.e., iPSC-CTLs, and investigated the suppression of SL9-specific iPSC-CTLs on viral replication and the protection of CD4+ T cells. A chimeric HIV-1, i.e., EcoHIV, was used to produce HIV replication in mice. We show that adoptive transfer of SL9-specific iPSC-CTLs greatly suppressed EcoHIV replication in the peritoneal macrophages and spleen in the animal model. Furthermore, we demonstrate that the adoptive transfer significantly reduced expression of PD-1 on CD4+ T cells in the spleen and generated persistent anti-HIV memory T cells. These results indicate that stem cell-derived viral Ag-specific CTLs can robustly accumulate in the local tissues to suppress HIV replication and prevent CD4+ T cell exhaustion through reduction of PD-1 expression.

Chimeric antigen receptor (CAR) T (CAR-T) cell transfer has made great success in hematological malignancies, but only shown a limited effect on solid tumors. One of the major hurdles is the poor persistence of infused cells derived from ex vivo activation/expansion and repeated antigen encounter after re-infusion. Bcl-xL has been demonstrated to play an important role on normal T cell survival and function as well as genetically engineered cells. In the current study, we developed a retroviral CAR construct containing a second-generation carcinoembryonic antigen (CEA)-targeting CAR with the Bcl-xL gene and tested the anti-CEA CAR-T cell immunotherapy for colorectal cancer. In vitro, the anti-CEA CAR-T cells destroyed CEA-expressing tumor cells and sustained survival. In vivo, adoptive cell transfer of anti-CEA CAR-T cells significantly enhanced the ability of the CAR-T cells to accumulate in tumor tissues, suppress tumor growth and increase the overall survival rate of tumor-bearing mice in a murine model of colorectal cancer. These results demonstrate a novel CAR-T platform that has the ability to increase the persistence of CAR-T cells in solid tumors through exogenous expression of persistent genes. The data provide a potentially novel approach to augment CAR-T immunotherapy for solid tumors.

Summary In T cell-based cancer immunotherapy, tumor antigen (Ag)-specific CD8+ cytotoxic T lymphocytes (CTLs) can specifically target tumor Ags on malignant cells. This promising approach drove us to adopt this strategy of T cell transfer (ACT)-based immunotherapy for chronic viral infections. Here, we describe the generation of hepatitis B virus (HBV) Ag-specific CTLs from induced pluripotent stem cells (iPSCs), i.e., iPSC-CTLs. Ag-specific iPSC-CTLs can target HBV Ag+ cells and infiltrate into the liver to suppress HBV replication in a murine model. For complete details on the use and execution of this protocol, please refer to Haque et al. (2020)., Graphical Abstract, Highlights • Generation of functional viral Ag-specific CTLs from iPSCs, i.e., iPSC-CTLs • Viral Ag-specific iPSC-CTLs suppress HBV replication in a murine model • Adoptive transfer of viral Ag-specific iPSC-CTLs induces persistent α-HBV immunity • Adoptive transfer of viral Ag-specific iPSC-CTLs produces antiviral cytokines, In T cell-based cancer immunotherapy, tumor antigen (Ag)-specific CD8+ cytotoxic T lymphocytes (CTLs) can specifically target tumor Ags on malignant cells. This promising approach drove us to adopt this strategy of T cell transfer (ACT)-based immunotherapy for chronic viral infections. Here, we describe the generation of hepatitis B virus (HBV) Ag-specific CTLs from induced pluripotent stem cells (iPSCs), i.e., iPSC-CTLs. Ag-specific iPSC-CTLs can target HBV Ag+ cells and infiltrate into the liver to suppress HBV replication in a murine model.

Autoimmune diseases are modern diseases characterized by destruction of own cells by host immune cells. The initial trigger for both systemic autoimmune disorders and organ-specific autoimmune disorders may involve the recognition of self or foreign molecules by innate immune determinants, which in turn trigger inflammatory responses and drive the engagement of previously undeveloped autoreactive T cells and B cells. Dysfunctional cytokine signaling of IL-2, IL-10, or TGF-β may also compromise the formation or function of Treg cells leading to severe autoimmunity. Scientific progress has led to a variety of new treatment approaches. Most of the biological approaches in clinical use rely upon the interference with proinflammatory cytokine function and signal transduction, inhibition of T cell activation, or depletion of B cells. These treatment regimens showed limited efficacy with systemic side effects. Induced pluripotent stem cells (iPSCs) can be generated from any somatic cells and can be differentiated into all cell types, which provide potential benefit by the generation of autologous source to overcome immune rejection of therapeutic cells. iPSCs have been studied more extensively in neurodegenerative and autoimmune disorders and have shown encouraging outcomes. iPSC technology has been utilized with potential for regeneration of specific neural populations or for exerting an immunomodulatory effect. iPSC technology has created an excellent clinically safe treatment option for autoimmune diseases while also providing suitable models to study diseases and develop new therapies in vitro. Altogether, iPSCs have revolutionized the approach of personalized regenerative medicine and developmental biology and given an unparalleled opportunity in deciphering the etiology and designing a treatment regimen. Clinical trials with iPSC-derived therapeutic cells to treat severely debilitating degenerative diseases are underway to ignite the preliminary approach into reality for effective treatment without compromising patient safety.

Over the past decade, autophagy has emerged as a critical regulatory mechanism of the immune system through critically controlling various aspects of T cell biology and determining the fate of different T cell subsets. Autophagy maintains T cell development and survival by regulating the degradation of organelles and apoptotic proteins. The autophagic process also impacts the formation of memory T cells. Alteration of autophagy in T cells may lead to a variety of pathological conditions such as inflammation, autoimmune diseases and cancer. In this review, we discuss how autophagy impacts T cell differentiation, survival and memory, and its implication in immunotherapy for various diseases.

Summary The viral antigen (Ag)-specific CD8+ cytotoxic T lymphocytes (CTLs) derived from pluripotent stem cells (PSCs), i.e., PSC-CTLs, have the ability to suppress hepatitis B virus (HBV) infection. After adoptive transfer, PSC-CTLs can infiltrate into the liver to suppress HBV replication. Nevertheless, the mechanisms by which the viral Ag-specific PSC-CTLs provoke the antiviral response remain to be fully elucidated. In this study, we generated the functional HBV surface Ag-specific CTLs from the induced PSC (iPSCs), i.e., iPSC-CTLs, and investigated the underlying mechanisms of the CTL-mediated antiviral replication in a murine model. We show that adoptive transfer of HBV surface Ag-specific iPSC-CTLs greatly suppressed HBV replication and prevented HBV surface Ag expression. We further demonstrate that the adoptive transfer significantly increased T cell accumulation and production of antiviral cytokines. These results indicate that stem cell-derived viral Ag-specific CTLs can robustly accumulate in the liver and suppress HBV replication through producing antiviral cytokines., Graphical Abstract, Highlights • Generation of functional viral Ag-specific CTLs from iPSCs, i.e., iPSC-CTLs • Viral Ag-specific iPSC-CTLs suppress HBV replication in a murine model • Adoptive transfer of viral Ag-specific iPSC-CTLs generates persistent α-HBV T cells • Adoptive transfer of viral Ag-specific iPSC-CTLs produces antiviral IFN-γ and TNF-⍺, Immunology; Functional Aspects of Cell Biology

Pancreatic ductal adenocarcinoma (PDAC) is an incurable cancer resistant to traditional treatments, although a limited number of early-stage patients can undergo radical resection. Immunotherapies for the treatment of haematological malignancies as well as solid tumours have been substantially improved over the past decades, and impressive results have been obtained in recent preclinical and clinical trials. However, PDAC is likely the exception because of its unique tumour microenvironment (TME). In this review, we summarize the characteristics of the PDAC TME and focus on the network of various tumour-infiltrating immune cells, outlining the current advances in PDAC immunotherapy and addressing the effect of the PDAC TME on immunotherapy. This review further explores the combinations of different therapies used to enhance antitumour efficacy or reverse immunodeficiencies and describes optimizable immunotherapeutic strategies for PDAC. The concordant combination of various treatments, such as targeting cancer cells and the stroma, reversing suppressive immune reactions and enhancing antitumour reactivity, may be the most promising approach for the treatment of PDAC. Traditional treatments, especially chemotherapy, may also be optimized for individual patients to remodel the immunosuppressive microenvironment for enhanced therapy.

Heat shock proteins (HSPs) are highly conserved molecular chaperones with divergent roles in various cellular processes. The HSPs are classified according to their molecular size as HSP27, HSP40, HSP60, HSP70, and HSP90. The HSPs prevent nonspecific cellular aggregation of proteins by maintaining their native folding energetics. The disruption of this vital cellular process, driven by the aberrant expression of HSPs, is implicated in the progression of several different carcinomas. Many HSPs are also actively involved in promoting the proliferation and differentiation of tumor cells, contributing to their metastatic phenotype. Upregulation of these HSPs is associated with the poor outcome of anticancer therapy in clinical settings. On the other hand, these highly expressed HSPs may be exploited as viable immunotherapeutic targets for different types of cancers. This review discusses recent advances and perspectives on the research of HSP-based cancer immunotherapy.

In the present report, composite membranes (CM) were prepared from a resin, derived from phenol and formaldehyde as matrix and mesoporous silica SBA-15 as filler on the clay-alumina tubular support. Membrane matrix was formed by in-situ cross-linking of phenol (P) and formaldehyde (F) at room temperature. The structural changes of membrane matrix with curing time influence the final properties of the membranes. The surface morphology and the filler-polymer interaction i.e. surface silanol group of SBA-15 and –CH 2 OH/–OH group of P–F resin of composite membrane were established by FESEM and Fourier transformation of infrared spectroscopy (FT-IR) and X-ray Photoelectron Spectroscopy (XPS) respectively. Thermal stability of the membranes was characterized by Thermogravimetric analysis (TGA) and Differential Thermal Analysis (DTA). Finally, the permeability and the separation efficiencies of the developed composite membranes were studied in details with varying curing time of resin at room temperature, feed pressure and different gas compositions for CO 2 /H 2 and CO 2 /CH 4 mixture gases. For CO 2 /H 2 ‘reverse selectivity’ was observed with the membrane. The highest separation factor of CO 2 /H 2 and CO 2 /CH 4 were achieved upto 16.6 and 62.4 for 67:33 (CO 2 /H 2 ) and 45:55 (CO 2 /CH 4 ) feed composition respectively. The room temperature cross-linking of P–F resin modified the interfacial defect of membrane and its effect on the CO 2 separation efficiencies is the unique observation of the present work. The obtained permeability and selectivity values of different gases were surpassing the 2008 Robeson upper bound line.

Compromise in adherens junctions (AJs) is associated with several chronic inflammatory diseases. We reported previously that Janus kinase 3, a non-receptor tyrosine kinase, plays a crucial role in AJ formation through its interaction with β-catenin. In this report, we characterize the structural determinants responsible for Jak3 interactions with β-catenin and determine the functional implications of previously unknown tyrosine residues on β-catenin phosphorylated by Jak3. We demonstrate that Jak3 autophosphorylation was the rate-limiting step during Jak3 trans-phosphorylation of β-catenin, where Jak3 directly phosphorylated three tyrosine residues, viz. Tyr30, Tyr64, and Tyr86 in the N-terminal domain (NTD) of β-catenin. However, prior phosphorylation of β-catenin at Tyr654 was essential for further phosphorylation of β-catenin by Jak3. Interaction studies indicated that phosphorylated Jak3 bound to phosphorylated β-catenin with a dissociation constant of 0.28 μm, and although both the kinase and FERM (Band 4.1, ezrin, radixin, and moesin) domains of Jak3 interacted with β-catenin, the NTD domain of β-catenin facilitated its interactions with Jak3. Physiologically, Jak3-mediated phosphorylation of β-catenin suppressed EGF-mediated epithelial–mesenchymal transition and facilitated epithelial barrier functions by AJ localization of phosphorylated β-catenin through its interactions with α-catenin. Moreover, loss of Jak3-mediated phosphorylation sites in β-catenin abrogated its AJ localization and compromised epithelial barrier functions. Thus, we not only characterize Jak3 interaction with β-catenin but also demonstrate the mechanism of molecular interplay between AJ dynamics and EMT by Jak3-mediated NTD phosphorylation of β-catenin.

PURPOSE OF REVIEW: Type 1 diabetes (T1D) is an autoimmune disease in which the immune cells selectively destroy the pancreatic beta (β) cells and results in the deficiency of insulin production. The optimal treatment strategy for T1D should be preventing of β-cell destruction in the pancreas. The purpose of this review is to discuss the immunological therapeutic mechanisms that will help to understand the development and control of β-cell destruction. The review also presents a novel method for development of autoantigen (Ag)-specific regulatory T cells (Tregs) for T1D immunotherapy. RECENT FINDINGS: Pancreatic-resident Tregs have the ability to dramatically suppress hyperactive immune cells. Islet cell transplantation is another attractive approach to replace the failed β cells. Due to the limited source of islet cells, research is going on in the use of animal cells and adult stem cells that may be derived from the patient’s own body to produce β cells for transplantation. SUMMARY: The mechanism behind the pancreatic β-cell destruction is largely unknown. In this review, a novel approach for the generation of tissue-associated Tregs from stem cells is considered. The stem cell-derived tissue-associated Tregs have the ability to home to the damaged pancreas to prevent the destruction. The review also provides new insights on the mechanism on how these suppressive immune cells protect the pancreas from the destruction of autoimmune cells. A novel method to develop functional auto Ag-specific Tregs that are derived from induced pluripotent stem cells (iPSCs), i.e., iPSC-Tregs, is discussed. Adoptive transfer of the iPSC-Tregs can substantially suppress T1D development in a murine model.

Intestinal microbiota harbors trillions of microbes that differ in composition among species and individuals. It serves as a checkpoint for pathogenic infections and has a synergistic role in influencing overall human health. Dietary habits, pathogen infection, lifestyle, and host's genotype-associated disorders disrupt the nature of host–commensal interaction called gut dysbiosis. The intestinal blood content and pathogens are received by liver from gut triggering liver activity. Thus, alterations in intestinal microflora play a key role in impairing the gut-liver axis, thereby affecting hepatic health. This chapter will provide an overview of gut microbiome, gut barrier function, and the potential role of gut microbes and pathogens in acute and chronic liver disease. Ultimately, understanding of these mechanisms will provide an insight into existing intervention strategies for treatment and prognosis of liver pathobiology.

Salmonella enterica serovars Enteritidis (S. Enteritidis) is one of the most common causes of food borne illness. Bacterial growth environment plays an important role in regulating gene expression thereby affecting the virulence profile of the bacteria. Different foods present diverse growth conditions which may affect the pathogenic potential of the bacteria. In the present study, the effect of food environments on the pathogenic potential of S. Enteritidis has been evaluated. S. Enteritidis was grown in different foods e.g. egg white, peanut butter and milk, and virulent phenotypes were compared to those grown in Luria Bertani broth. In-vivo experiments in C57BL/6 mice revealed S. Enteritidis grown in egg white did not induce significant (p0.001) production of proinflammatory cytokines in mice and were unable to cause colitis despite efficient colonization in cecum, mesenteric lymph node, spleen and liver. Further studies revealed that bacteria grown in LB activated MAP Kinase and NFκB pathways efficiently, while those grown in egg white poorly activated the above pathways which can account for the decreased production of proinflammatory cytokines. qRT PCR analysis revealed SPI-1 effectors were downregulated in bacteria grown in egg white. Interestingly, bacteria grown in egg white showed reversal of phenotype upon change in growth media to LB. Additionally, bacteria grown in milk and peanut butter showed different degrees of virulence in mice as compared to those grown in LB media. Thus, the present study demonstrates that, S. Enteritidis grown in egg white colonizes systemic sites without causing colitis in a mouse model, while bacteria grown in milk and peanut butter show different pathogenicity profiles suggesting that food environments significantly affect the pathogenicity of S. Enteritidis.

In this work, the high-quality ilmenite (FeTiO3) membranes were grown on the clay-Al2O3 support tube from microemulsion-derived FeTiO3 powders by dip-coating method. FeTiO3 powders were synthesized by reverse microemulsion method, using cyclohexane, Tween-80, Brij 30, Triton X-100, and n-butanol at different composition. The membranes as well as powders were finally characterized by removal of red dye from a simulated wastewater of textile industry in same conditions. The membrane separation process demonstrates the removal of dye coupled with permeation of water simultaneously, and the overall process may have great potential for wide industrial application.

We report a unique alloy–dealloy phenomenon of Au–Ag nanoparticles inside SiO2 films with clear plasmonic modifications between the absorptions of Ag (∼415 nm) and Au (∼524 nm). An Au–Ag (1 : 1) alloy nanoparticles (average size: 4.5 nm)-incorporated transparent SiO2 film is prepared on a glass substrate using mercaptosuccinic acid capped Au nanoparticles and Ag+ co-doped hybrid sol. The Au–Ag (1 : 1) alloy-originated plasmon band (465 ± 1 nm) is gradually red-shifted with increasing temperature (50 to 400 °C) due to the partial oxidation of Ag, causing a systematic modification of the alloy composition. The 1 : 1 alloy, however, reverted very quickly, showing its original plasmon band in the presence of a small amount of H2 due to re-reduction of the oxidized Ag and instantaneous re-alloying. During the Ag oxidation, the Si–OH groups associated with the embedded SiO2 matrix exchange Ag+ to form Ag–O–Si linkages; they subsequently release Ag very quickly in H2 and dissolve again into the parent alloy. As a result, the films exhibit reversible and rapid optical changes while cycling in 0.1% to 1% H2 (balance Ar) and air in the temperature range from 50 to 400 °C. This unique reversible alloy–dealloy phenomenon clearly demonstrates the mechanism of plasmonic modification associated with Au–Ag nanoparticles embedded in the sol–gel SiO2 film matrix.

The autoantigen-specific Tregs from pluripotent stem cells (PSCs), i.e., PSC-Tregs, have the ability to suppress autoimmunity. PSC-Tregs can be programmed to be tissue associated and to infiltrate into local inflamed tissues to suppress autoimmune responses after adoptive transfer. Nevertheless, the mechanisms by which the autoantigen-specific PSC-Tregs suppress the autoimmune response remain to be fully elucidated. In this study, we generated functional autoantigen-specific Tregs from the induced PSC (iPSCs), i.e., iPSC-Tregs, and investigated the underlying mechanisms of autoimmunity suppression by these Tregs in a type 1 diabetes (T1D) murine model. A double-Tg mouse model of T1D was established in F(1) mice, in which the first generation of RIP-mOVA Tg mice that were crossed with OT-I T cell receptor (TCR) Tg mice was challenged with vaccinia viruses expressing OVA (VACV-OVA). We show that adoptive transfer of OVA-specific iPSC-Tregs greatly suppressed autoimmunity in the animal model and prevented the insulin-secreting pancreatic β cells from destruction. Further, we demonstrate that the adoptive transfer significantly reduced the expression of ICAM-1 in the diabetic pancreas and inhibited the migration of pathogenic CD8(+) T cells and the production of the proinflammatory IFN-γ in the pancreas. These results indicate that the stem cell–derived tissue-associated Tregs can robustly accumulate in the diabetic pancreas, and, through downregulating the expression of ICAM-1 in the local inflamed tissues and inhibiting the production of proinflammatory cytokine IFN-γ, suppress the migration and activity of the pathogenic immune cells that cause T1D.

The Mur enzymes of the peptidoglycan biosynthesis pathway constitute ideal targets for the design of new classes of antimicrobial inhibitors in Gram-negative bacteria. We built a homology model of MurD of Salmonella typhimurium LT2 using MODELLER (9v12) software. 'The homology model was subjected to energy minimization by molecular dynamics (MD) simulation study with GROMACS software for a simulation time of 20 ns in water environment. The model was subjected for virtual screening study from the Zinc Database using Dockblaster software. Inhibition assay for the best inhibitor, 3-(amino methyl)-n-(4-methoxyphenyl) aniline, by flow cytometric analysis revealed the effective inhibition of peptidoglycan biosynthesis. Results from this study provide new insights for the molecular understanding and development of new antibacterial drugs against the pathogen.

A simple method of pore modification coupled with the removal of surface defects of a deca-dodecasil-rhombohedral (DDR) zeolite membrane has been developed via carbon (C) deposition. Carbon deposition was achieved by controlled decomposition of the structure-directing agent (SDA), 1-adamantanamine (1-ADA), into the membrane pore surface. Membranes that were synthesized with and without pore modification were characterized by thermal analysis, X-ray diffraction analysis, infrared spectroscopy, transmission electron microscopy, and Raman spectroscopy. Finally, the performance of the membranes was evaluated by permeance studies. The non-wetting characteristics of C had imparted hydrophobicity to the membrane pores, leading to enhanced permeability of the gas mixture. Modified membranes were shown to offer a relatively high hydrogen permeance of 13.47 × 10−7 mol m−2 s−1 Pa−1. The selectivity of H2/CO2 was 4.9 based on single gas permeation and the separation factor increased to 8.5 for a H2–CO2 gas mixture at room temperature. In the light of these findings, the current technique is proposed to be useful for making a defect-free C-loaded membrane in a single step with high separation selectivity and permeability in tandem. This is the novelty that was achieved.

Aim: To investigate molecular aspects of the antibacterial effect of size-dependent core-shell intrinsic defects of nanocrystalline ZnO synthesized through high energy ball milling technique. Materials & methods: Mechanically synthesized and characterized 7, 10 and 15 h milled ZnO nanoparticles were evaluated for antibacterial activity with molecular investigation by computational molecular docking. Results: Synthesized ZnO nanoparticles displayed shrinkage of core and increase of shell with reduction in size of bulk ZnO particles from 250 to 80, 40 and 20 nm and increase in zeta potential up to -19 mV in 7, 10 and 15 h nano ZnO. Antibacterial activity was found increased with decrease in size due to increased reactive oxygen species and membrane damage in bacteria. Conclusion: Synthesized nano ZnO exhibit size-dependent antibacterial action as consequences of interactions with cell membrane proteins via hydrogen bond interaction with amino acid residues followed by internalization, membrane depolarization and induction of reactive oxygen species generation.

OBJECTIVE Prevention or induction of metabolic disorders and obesity depend on estrogen signaling and/or exogenous factors, such as mineral content in diet. The protective effects of a Portuguese natural mineral-rich water against the induction of metabolic syndrome in fructose-fed male Sprague-Dawley rats have been reported. The present study was designed to assess the impact of this mineral-rich water on fructose-fed estrogen-deficient female Sprague-Dawley rats. METHODS Ovariectomized rats had access to tap (TWO) or mineral-rich (MWO) waters, with and without 10% fructose (10-wk treatment). A sham-operated (tap water supplied) group was included and each of the five groups included six rats. Plasma biochemical and metabolic parameters were evaluated by routine clinical measurements. Western blotting was used to assess hepatic protein expression of sirtuins (Sirt) 1 and 3, phosphorylated AMP-activated protein kinase-α (p-AMPKα), peroxisome proliferator-activated receptor gamma coactivator-1-α (PGC1α), glucocorticoid receptor, and 11beta-hydroxysteroid dehydrogenase type 1 (11βHSD1). RESULTS Ovariectomy increased plasma total cholesterol (46%/P

In this work, the high quality Pd/SAPO 34 membranes were grown on the support using a secondary (seeded) growth hydrothermal technique followed by insertion of 11-mercaptoundecanoic acid capped palladium (MUA-Pd) nanoparticles (NPs) to the membrane surface. For this, first, the indigenous low cost clay–alumina support was treated with poly diallyldimethylammonium chloride (PolyDADMAC) polymer, and subsequently, a seed layer of SAPO 34 crystals was deposited homogeneously in a regular orientation. Since PolyDADMAC is a high charge density cationic polymer, it assisted in reversing the charge of the support surface and produced an attractive electrostatic interaction between the support and zeolite crystals. This may facilitate the zeolite grain orientation in the synthesized membrane layer. Here, the Pd NPs were deposited in the membrane matrix by a simple dip-coating method. After thermal treatment of the Pd/SAPO 34 membrane, the defects were formed because of the removal of the structure-directing agent ...

Eukaryotic elongation factor-2 kinase (eEF-2K) is a conserved protein kinase which associates multiple upstream signals with protein synthesis and cellular metabolism. Reprogramming of the cellular metabolic microenvironment in T-lymphocytes is a hallmark of several immune-pathophysiological conditions. However, the role of eEF-2K in T-cell metabolism and signaling remain unclear. The aim of the present study was to unravel the critical role of eEF-2K in regulation of cytotoxic CD8+T-lymphocytes (CTLs). Primary CTLs from wild type (WT) and eEF-2K knock-out (KO) C57BL/6 mice were stimulated with anti-CD3 and CD28 antibodies and examined for their survival, proliferation and cytokine production. The cellular metabolism was analyzed using Seahorse Extracellular Flux analyzer. A combination of western blotting and flowcytometric analysis was used to assess the key mediators of the central metabolic Akt-PI3k-mTOR pathway. Following activation, eEF-2K KO CTLs had significantly reduced survival on Day 4 (4-fold change; p Collectively, our results identify that eEF-2K not only plays a crucial role in CTL activation and signaling, but also regulates senescence. These findings may be useful in designing novel strategies by targeting eEF-2K-based immunotherapy.

In this work, DDR zeolites were synthesized by sonochemical method without the application of any heat energy at room temperature. The synthesized zeolites were characterized by X-ray diffraction (XRD), infrared (IR) spectral analysis, and field-emission scanning electron microscopy (FESEM). XRD and IR results showed that phase pure DDR zeolite was started to form at room temperature after 24 h of aging and completed the formation after 5 d of aging. The Brunauer–Emmett–Teller (BET) surface area of the powder was found to be 202 m 2 /g. The FESEM micrograph and elemental analysis showed that desired atomic ratio of the DDR zeolite was obtained after 5 days of synthesis.

NaP zeolite nano crystals were synthesized by sonochemical method at room temperature with crystallization time of 3 h. For comparison, to insure the effect of sonochemical method, the hydrothermal method at conventional synthesis condition, with same initial sol composition was studied. NaP zeolites are directly formed by ultrasonic treatment without the application of autogenous pressure and also hydrothermal treatment. The effect of ultrasonic energy and irradiation time showed that with increasing sonication energy, the crystallinity of the powders decreased but phase purity remain unchanged. The synthesized powders were characterized by XRD, IR, DTA TGA, FESEM, and TEM analysis. FESEM images revealed that 50 nm zeolite crystals were formed at room temperature by using sonochemical method. However, agglomerated particles having cactus/cabbage like structure was obtained by sonochemical method followed by hydrothermal treatment. In sonochemical process, formation of cavitation and the collapsing of bubbles produced huge energy which is sufficient for crystallization of zeolite compared to that supplied by hydrothermal process for conventional synthesis. With increasing irradiation energy and time, the crystallinity of the synthesized zeolite samples increased slightly.

The formation and growth of Silicoaluminophosphate (SAPO 34) zeolite membrane was developed on the low cost indigenous clay–alumina tubular substrate modified by cationic polymer as an intermediate layer. The amino (–NH2) and hydroxyl (–OH) groups on the backbone of polymer are responsible for hydrogen bonding between zeolite surface and support surface. Seeding of the support surface accelerates the zeolite crystallization and enhances the formation of homogenous SAPO 34 membrane layer. The seeds were synthesized by hydrothermal process and used to provide nucleation for the membrane growth. The synthesized membranes along with seed crystals were characterized by XRD, FTIR, FESEM, TEM and EDAX analysis. The performance of the membrane formed was evaluated by single gas as well as mixture gas permeation measurement for H2 and CO2. The H2/CO2 separation selectivity of the membrane increased upto 5.88 at room temperature which is more than reported values and the reproducibility towards gas permeation and selectivity of SAPO 34 membrane shows excellent result.

SAPO 34 zeolite membranes were prepared on a tubular mullite support. Before membrane preparation, the support surfaces were coated with seed crystals. Seeds particles were prepared by hydrothermal synthesis. Before seeding, the substrates were treated with polyvinylpyrrolidone (PVP) to orient the seeds. Both the treated and untreated supports were seeded, and membranes were synthesized on those support tubes by ex situ hydrothermal method. The PVP molecule exists in the two resonance structures. Hence the acylamino bond –N+= C–O-– acts as intermediate linker between support surface and seed surface. Due to charge interaction, the seed crystals were anchored strongly to support surface. The synthesized membranes along with seed crystals were characterized by XRD, FESEM, and EDAX analysis. The single-gas permeation with CO2and H2was investigated. Up to 5 bar of feed pressure, the permselectivity of CO2and H2was as high as 4.2.

The formation stage of SAPO 34 zeolite membrane on a tubular mullite support has been investigated. XRD, FESEM, IR and EDAX analysis techniques were used to explain the changes in crystallization stages as well as formation of membrane on tubular clay–alumina support with time. From the studies, the evidence tends to support that crystallization of SAPO 34 started from initial gel, proceed through cluster formation and accumulation followed by segregation and crystallization process. The study also showed the gradual incorporation of Si into AlPO 4 phase and form cubical CHA phase after 120 h of synthesis time. Single gas permeation of CO 2 and H 2 showed that upto 300 KPa of feed pressure permeability of CO 2 is more than that of H 2 . But at higher pressure the results show the reverse trend, flux is more for H 2 . This may be due to more adsorption of CO 2 on SAPO 34 surface and less desorption from the surface than hydrogen with increasing pressure. The selectivity of H 2 /CO 2 increases from 0.85 to 2.67.

Hematopoietic stem cells (HSCs) yield both the myeloid and lymphoid lineages of blood cells and can be reprogrammed into tumor antigen (Ag)-specific CD8+ cytotoxic T lymphocytes (CTLs) to prevent tumor growth. However, the optimal approach for differentiating tumor Ag-specific CTLs from HSCs, such as HSC-CTLs, remains elusive. In the current study, we showed that a combination of genetic modification of HSCs and in vivo T cell development facilitates the generation of Ag-specific CTLs that suppressed tumor growth. Murine HSCs, which were genetically modified with chicken ovalbumin (OVA)-specific T cell receptor, were adoptively transferred into recipient mice. In the following week, mice were administered with intraperitoneal injections of an agonist &alpha, Notch 2 antibody and cytokines (rFlt3L and rIL-7) three times. After another two weeks, mice received a subcutaneous inoculation of B16-OVA melanoma cells that express OVA as a surrogate tumor Ag, before the anti-tumor activity of HSC-derived T cells was assessed. OVA-specific CTLs developed in vivo and greatly responded to OVA Ag stimulation ex vivo. In addition, mice receiving genetically modified HSCs and in vivo priming established anti-tumor immunity, resulting in the suppression of tumor growth. These results reported in this present study provide an alternative strategy to develop protective cancer vaccines by using genetically modified HSCs.

In this work, the high quality Pd/SAPO 34 membranes were grown on the support using a secondary (seeded) growth hydrothermal technique followed by insertion of 11-mercaptoundecanoic acid capped palladium (MUA-Pd) nanoparticles (NPs) to the membrane surface. For this, first, the indigenous low cost clay-alumina support was treated with poly diallyldimethylammonium chloride (PolyDADMAC) polymer, and subsequently, a seed layer of SAPO 34 crystals was deposited homogeneously in a regular orientation. Since PolyDADMAC is a high charge density cationic polymer, it assisted in reversing the charge of the support surface and produced an attractive electrostatic interaction between the support and zeolite crystals. This may facilitate the zeolite grain orientation in the synthesized membrane layer. Here, the Pd NPs were deposited in the membrane matrix by a simple dip-coating method. After thermal treatment of the Pd/SAPO 34 membrane, the defects were formed because of the removal of the structure-directing agent (SDA) from the zeolite pores but the presence of Pd NPs, which were entrapped inside the nonzeolitic pores and clogged the defects of the membrane. Field emission scanning electron microscopy (FESEM) and elemental mapping of the membrane cross-section confirmed that most of the Pd NPs were deposited at the interface of the membrane and the support layer which may increase the membrane efficiency, i.e., separation factor, as well as permeability of H2 through the membrane. As the membrane structure was associated with the oriented crystal, the pores were more aligned and permeation adequacy of H2 through the membrane enhanced. These membranes have a relative hydrogen permeance of 14.8 × 10(-7) mol·m(-2)·s(-1)·Pa(-1). The selectivity of H2/CO2 based on single gas permeation was 10.6, but for the mixture gas (H2/CO2 55:45), the H2/CO2 mixture separation factor increased up to 20.8 at room temperature. It is anticipated that this technique may be useful for making a defect free membrane and also a hydrogen selective Pd loaded membrane with lower cost (as the quantity of Pd is low) which can be utilized for a "clean energy" related application.

Lactobacillus strains have been shown to adhere to the mucosal components of intestinal epithelial cells. However, established in vitro adhesion assays have several drawbacks in assessing the adhesion of new Lactobacillus strains. The present study aimed to compare the adhesion of four different Lactobacillus strains and select the most adherent microbe, based on in silico approach supported by in vitro results. The mucus-binding proteins in Lactobacillus acidophilus, L. plantarum, L. brevis and L. fermentum were identified and their capacities to interact with intestinal mucin were compared by molecular docking analysis. Lactobacillus acidophilus had the maximal affinity of binding to mucin with predicted free energy of -6.066 kcal mol(-1) Further, in vitro experimental assay of adhesion was performed to validate the in silico results. The adhesion of L. acidophilus to mucous secreting colon epithelial HT-29 MTX cells was highest at 12%, and it formed biofilm with maximum depth (Z = 84 μm). Lactobacillus acidophilus was determined to be the most adherent strain in the study. All the Lactobacillus strains tested in this study, displayed maximum affinity of binding to MUC3 component of mucus as compared to other gastrointestinal mucins. These findings may have importance in the design of probiotics and health care management.