Exploring peptidomes of by-products generated during chhurpi production using Lactobacillus delbrueckii WS4 for identification of novel bioactive peptides.

The considerable value of whey is evident from its significant potential applications and contributions to the functional food and nutraceutical market. The by-products were individually obtained during functional chhurpi and novel soy chhurpi cheese production using defined lactic acid bacterial strains of Sikkim Himalaya's traditional chhurpi. Hydrolysis of substrate proteins by starter proteinases resulted in a comparable peptide content in whey and soy whey which was associated with antioxidant and ACE inhibition potential. Peptidome analysis of Lactobacillus delbrueckii WS4 whey and soy whey revealed the presence of several bioactive peptides including the multifunctional peptides PVVVPPFLQPE and YQEPVLGPVRGPFPIIV. In silico analyses predicted the antihypertensive potential of whey and soy whey peptides with strong binding affinity for ACE active sites. QSAR models predicted the highest ACE inhibition potential (IC50) for the ß-casein-derived decapeptide PVRGPFPIIV (0.95 µM) and the Kunitz trypsin inhibitor protein-derived nonapeptide KNKPLVVQF (16.64 µM). Chhurpi whey and soy whey can be explored as a valuable source of diverse and novel bioactive peptides for applications in designer functional foods development.

A study on incorporation of giloy (Tinosporacordifolia) for the development of shelf-stable goat milk based functional beverage.

Goat milk has achieved significant place in human diet owing to its enormous therapeutic properties. There exists a scope of value-addition of goat milk to potentiate its health benefits by incorporating herbs and plants. Giloy (Tinospora cordifolia), a traditional medicinal plant with rich bioactive composition, can enhance the bioactive properties and shelf-life of goat milk. To this end, a study was conducted to develop shelf-stable giloy-goat milk beverage (GGB) by adding debittered giloy juice to goat milk (GM) and analyse the detailed product profile including proximate composition, bioactive properties, sensory, rheological, and structural characterisation. GGB resulted in two-fold increase (P < 0.05) in antioxidant activity and total phenolic content, thus enhancing the bioactive properties of the beverage as compared to GM. Further, increase in the particle size of GGB was observed along with components interaction, which was confirmed by FTIR, scanning electron and fluorescent microscopy. Storage stability studies indicated that bioactive properties of GGB remained unaffected (P > 0.05) by the sterilization process up to 90 days and sensory characteristics were not compromised till 105 days of storage. Therefore, the developed GGB is considered to be a shelf-stable beverage that retains its bioactive and sensory properties even after sterilization, making it a promising functional dairy product.

Role of cytokine in malignant T-cell metabolism and subsequent alternation in T-cell tumor microenvironment.

T cells are an important component of adaptive immunity and T-cell-derived lymphomas are very complex due to many functional sub-types and functional elasticity of T-cells. As with other tumors, tissues specific factors are crucial in the development of T-cell lymphomas. In addition to neoplastic cells, T- cell lymphomas consist of a tumor micro-environment composed of normal cells and stroma. Numerous studies established the qualitative and quantitative differences between the tumor microenvironment and normal cell surroundings. Interaction between the various component of the tumor microenvironment is crucial since tumor cells can change the microenvironment and vice versa. In normal T-cell development, T-cells must respond to various stimulants deferentially and during these courses of adaptation. T-cells undergo various metabolic alterations. From the stage of quiescence to attention of fully active form T-cells undergoes various stage in terms of metabolic activity. Predominantly quiescent T-cells have ATP-generating metabolism while during the proliferative stage, their metabolism tilted towards the growth-promoting pathways. In addition to this, a functionally different subset of T-cells requires to activate the different metabolic pathways, and consequently, this regulation of the metabolic pathway control activation and function of T-cells. So, it is obvious that dynamic, and well-regulated metabolic pathways are important for the normal functioning of T-cells and their interaction with the microenvironment. There are various cell signaling mechanisms of metabolism are involved in this regulation and more and more studies have suggested the involvement of additional signaling in the development of the overall metabolic phenotype of T cells. These important signaling mediators include cytokines and hormones. The impact and role of these mediators especially the cytokines on the interplay between T-cell metabolism and the interaction of T-cells with their micro-environments in the context of T-cells lymphomas are discussed in this review article.

Detection of germline variants with pathogenic potential in 48 patients with familial colorectal cancer by using whole exome sequencing.

BACKGROUND: Hereditary genetic mutations causing predisposition to colorectal cancer are accountable for approximately 30% of all colorectal cancer cases. However, only a small fraction of these are high penetrant mutations occurring in DNA mismatch repair genes, causing one of several types of familial colorectal cancer (CRC) syndromes. Most of the mutations are low-penetrant variants, contributing to an increased risk of familial colorectal cancer, and they are often found in additional genes and pathways not previously associated with CRC. The aim of this study was to identify such variants, both high-penetrant and low-penetrant ones. METHODS: We performed whole exome sequencing on constitutional DNA extracted from blood of 48 patients suspected of familial colorectal cancer and used multiple in silico prediction tools and available literature-based evidence to detect and investigate genetic variants. RESULTS: We identified several causative and some potentially causative germline variants in genes known for their association with colorectal cancer. In addition, we identified several variants in genes not typically included in relevant gene panels for colorectal cancer, including CFTR, PABPC1 and TYRO3, which may be associated with an increased risk for cancer. CONCLUSIONS: Identification of variants in additional genes that potentially can be associated with familial colorectal cancer indicates a larger genetic spectrum of this disease, not limited only to mismatch repair genes. Usage of multiple in silico tools based on different methods and combined through a consensus approach increases the sensitivity of predictions and narrows down a large list of variants to the ones that are most likely to be significant.

Synthesis and in vitro antibacterial behavior of curcumin-conjugated gold nanoparticles.

Owing to the rise in multidrug-resistant bacterial diseases and the dwindling supply of newer antibiotics, it is crucial to discover newer compounds or modify current compounds for more effective antimicrobial therapies. According to reports, more than 80% of bacterial infections have been linked to bacterial biofilms. In addition to having antimicrobial properties, the hydrophobic polyphenol curcumin (Cur) also inhibits quorum sensing. The application of curcumin was constrained by its weak aqueous solubility and quick degradation. Over the past years, nanotechnology-based biomaterials with multi-functional characteristics have been engineered with high interest. The present study focused on the development of nano-biomaterials with excellent testifiers for bacterial infection in vitro. In this study, water dispersibility and stability of curcumin were improved through conjugation with gold nanoparticles. The successful synthesis of curcumin-conjugated gold nanoparticles (Cur-AuNPs) was confirmed using X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, and UV-vis absorbance spectroscopy. Transmission electron microscopy (TEM) revealed an average particle size of about 10-13 nm. The antibacterial characteristics in terms of the minimum inhibitory concentration (MIC) of Cur-AuNP treatments were found to be lowest than those with AuNPs and Cur treatments. The quantitative analysis revealed the superior antibacterial characteristics of Cur-AuNP-treated bacterial cells compared to the untreated samples. In addition, curcumin-conjugated AuNPs, produced more reactive oxygen species and increased the membrane permeability. Besides, the biocompatibility of Cur-AuNPs was also assessed quantitatively and qualitatively. Statistical analyses revealed the augmented MG-63 cell proliferation in Cur-AuNPs compared to those with Cur and AuNPs treatments. Overall, Cur-AuNPs exhibited enhanced antibacterial, and antibiofilm characteristics and cytocompatibility.

Insight to chromium homeostasis for combating chromium contamination of soil: Phytoaccumulators-based approach.

Chromium (Cr) is a naturally occurring, carcinogenic heavy metal that has become a pressing concern in recent decades for environmentalists. Due to high anthropogenic activities, the concentration of Cr has crossed the environmental threshold levels and consequently contaminated soil and water. The high solubility of Cr ions in the groundwater results in its high uptake by the plants leading to phytotoxicity and yield loss. The dearth of efficient and cost-effective treatment methods has resulted in massive chromium pollution. However, some phytoaccumulators capable of accumulating Cr in high amounts in their shoots and then performing their metabolic activity typically have been identified. Chromium bioremediation using phytoaccumulators is very contemplative due to its eco-friendly and cost-effective outcome. These accumulators possess several mechanisms, such as biosorption, reduction, efflux, or bioaccumulation, naturally or acquired to counter the toxicity of chromium. This review focuses on the detoxification mechanism of Cr by the phytoaccumulator species, their responses against Cr toxicity, and the scope for their application in bioremediation. Besides, Cr bioavailability, uptake, distribution, impairment of redox homeostasis, oxidative stress, and phytotoxicity imposed on the plants are also summarized. Further, the knowledge gap and prospects are also discussed to fill these gaps and overcome the problem associated with the real-time applicability of phytoaccumulator-based bioremediation.

Different Treatments for Sugarcane Juice Preservation.

This investigation aimed to optimize the time, pH, pressure, and temperature of sugarcane juice pasteurization and to develop a "ready to serve" bottled sugarcane juice with a high preservation efficiency. Fresh sugarcane juice was extracted from sugarcane genotype Co 89003, and beverage samples were collected using three different treatments: sulphitation of juice with the addition of potassium metabisulphite (KMS-25, 50, 100, and 150 ppm), acidification of juice (addition of citric acid, to reduce the pH of the juice to 4.8, 4.5, and 4.25), and steam treatment of the canes (5 min, 10, and 15 min at 7 psi). In all treatments, the juice was pasteurized in glass bottles @ 65 °C for 25 min and stored at low temperature (5 °C) in pre-sterilized glass bottles. Juice properties such as the ˚Brix, total sugar, pH, and total phenolic content decreased with storage, whereas the microbial count, titrable acidity, and reducing sugar content significantly increased during storage. The addition of KMS, citric acid, and the steam treatment reduced the browning of juice and maintained the color of juice during storage, by inhibiting the polyphenol oxidase enzyme activity, from 0.571 unit/mL to 0.1 unit/mL. Among the selected treatments, sugarcane juice with KMS (100 and 150 ppm) and steam treatment of the canes for 5 and 10 min at 7 psi showed the minimum changes in physico-chemical properties, sensory qualities, and restricted microbial growth. Thesulphitation treatment with pasteurization proved best for increasing the shelf life of sugarcane juice upto 90 days with refrigeration. Similarly, the steam-subjected cane juice (10 and 15 min at 7 psi) could be effectively preserved for upto 30 days with refrigeration, without any preservative.

Effect of probiotic fermentation on physico-chemical and nutritional parameters of milk-cereal based composite substrate.

Abstract: A dairy-cereal based composite substrate was prepared from whey-skim milk (60:40 v/v), germinated pearl millet flour (4.73% w/v) and liquid barley malt extract (3.27% w/v) and fermented using probiotic strain Lactobacillus acidophilus NCDC-13. Probiotic fermentation increased whiteness index, viscosity and water holding capacity of unfermented substrate. Fermentation caused a reduction in total solids, fat, ash and total dietary fibre content and increment in protein content. Fermentation brought a highly significant reduction in phytic acid (78%) and polyphenol (46%) content. The protein and starch digestibility increased significantly. The HCl- extractability of Ca, Fe, Mg and Zn of unfermented substrate was 32%, 25%, 64% and 17% respectively, which increased to 73%, 50%, 83% and 65% respectively after fermentation. Fermentation resulted in 77% decrease in phytate P as % of total P and significant increase in free P. The current investigation revealed that probiotic fermentation improved nutritional attributes of the composite substrate substantially. The low cost nutritionally enriched probiotic substrate can be utilized for preparation of a wide range of low- cost probiotic foods to address malnutrition and enhance immunity of common population.

Transcription factor 4 expression and correlation with tumor progression in gallbladder cancer.

Background: Dysregulation in Wnt/ß-catenin signaling has been associated with the initiation and metastasis of cancer cells. Transcription factor 4 (TCF4) (also named as transcription factor 7-like 2) is a key transcriptional factor of the Wnt signaling pathway, which, when interact with ß-catenin activates Wnt genes which plays an essential role in tumor development. The expression pattern and clinical significance of TCF4 in gallbladder cancer (GBC) are not yet established. Aims: This study was performed to assess the expression pattern of TCF4 in GBC tissue and attempted to correlate its expression with different clinicopathological parameters. Materials and Methods: The study was conducted on 33 surgically resected specimens of gallbladder carcinoma (GBC) and 12 cases of chronic cholecystitis (CC) as control, which had been confirmed from histology. The expression of TCF4 was performed by the reverse transcription polymerase chain reaction and immunohistochemistry. Results: Relative mRNA expression levels of ß-catenin and TCF4 in GBC tissues were significantly (P < 0.05) higher than in CC samples. TCF4 protein expression was observed in 81.82% (27/33) GBC cases. Specifically, among GBC samples, 21.21% (7/33) was graded as strongly positive, 60.61% (20/33) graded as moderately positive, whereas 18.18% (6/33) graded as negative. All 12 CC samples graded as negative. Overall, TCF4 expression in GBC tissues was statistically significant over CC samples (P < 0.05). Moreover, we observed that TCF4 expression was significantly higher (P < 0.05) in high tumor grades than low grade, higher (P < 0.05) in Stage 2 and Stage 3 than Stage 1. Conclusion: The present study suggests that TCF4 may exert an oncogenic role in the progression of GBC and may serve as a new potential candidate biomarker for tumor progression, and it might be a potential therapeutic target against GBC.

STUDY OF EFFECTIVENESS OF SHELTERING FROM RADIATION EXPOSURE DUE TO ACCIDENTAL RELEASE OF RADIOACTIVITY.

Sheltering is one of the important protective actions as a part of emergency response during the early phases of an accident-mostly precautionary in nature. Sheltering via structural shielding reduces the direct exposure from plume/cloud shine and ground shine doses as well as the airborne radioactivity concentration and hence inhalation dose. The present study was aimed at estimating the shielding factor in the case of Indian houses for external exposure pathways using the FLUKA Monte Carlo based radiation transport code. Furthermore, the dose reduction factor due to exposure from the inhalation pathway was estimated using an indoor aerosol model. These factors were assessed for the three major types of dwellings, and they provided important inputs for decision-making for sheltering or evacuation in case of any sudden release of radionuclides into the environment.

Optimization of Tobacco Rattle Virus (TRV)-Based Virus-Induced Gene Silencing (VIGS) in Tomato.

Unveiling of full genome sequence of tomato demands significant advances of tomato functional genomics. Virus-induced gene silencing (VIGS) is a well explored functional genomics tool in plant biology that exploits post transcriptional gene silencing to downregulate a desired gene. Although VIGS provides an easy and highly efficient platform to study plant gene function through reverse genetics approach, currently VIGS is more efficient in model plants like Nicotiana benthamiana, which further justifies the urgent need of a highly efficient, reliable, and reproducible VIGS protocol in crop plants such as tomato. In this chapter, we have detailed an optimized Tobacco rattle virus (TRV)-based VIGS protocol in tomato.

Port-Site Metastases in Incidentally Detected Carcinoma Gall Bladder: Role of 18F-FDG PET/CT and Patient Outcome.

BACKGROUND: Incidentally detected gall bladder carcinoma (IGBC) is occasionally encountered after cholecystectomy for benign gall bladder disease. Rarely these patients may present with port-site metastases (PSM) in follow-up. We retrospectively assessed the role of 18F-fluorodeoxyglucose positron emission tomography-computed tomography (FDG PET/CT) in them. We aimed to determine the clinical outcome and correlate the survival based on FDG PET/CT findings. MATERIAL AND METHOD: We did a single-center retrospective study in IGBC patients presented with histopathologically proven PSM. FDG PET/CT was done for restaging. The metastatic burden was evaluated. Survival after PSM and overall survival (OS) were assessed in different disease groups. RESULTS: A total of 22 patients (51.86 ± 12.04 years) were included in the study. The mean interval between LC and PSM was 336.7 ± 501.9 days. On FDG PET/CT, isolated single PSM was noted in 21 (95.5%) patients. Apart from PSM, abdominal lymph nodes, GB fossa lesion, and liver metastases were noted in the 10 (45.5%), 6 ( 27.3%), and 7 (31.8%) patients, respectively. FDG PET/CT revealed intra-abdominal disease and visceral metastasis in 11 (50%) and 10 (45.5%) patients. Nineteen (86.4%) patients died in follow-up with a median survival of 328.50 (225.25-804.0) day after PSM. Median OS after LC was 687.00 (362.50-1047.0) days. Patients with PSM plus disease (432.0 days, 255.9-608.1), extra-abdominal metastases (400.0 days, 316.9-483.1), and visceral metastases (400.0 days, 296.2-503.8) had significantly lower OS compared to isolated PSM (1763 days, p = 0.013), the intra-abdominal disease (996.0 days, 753.2-1238.8, p = 0.0340), and non visceral metastasis (807.0 days, 467.5-1146.4, p = 0.037). IGBC presenting as PSM is a distinct clinical entity. FDG PET/CT as functional imaging evaluates the local and metastatic disease burden. There is significantly lower survival in patients with PSM plus disease, extra-abdominal metastases, or visceral metastases. FDG PET/CT was an important prognostic marker in these patients and could help in management.

Growth hormone induces mitotic catastrophe of glomerular podocytes and contributes to proteinuria.

Glomerular podocytes are integral members of the glomerular filtration barrier in the kidney and are crucial for glomerular permselectivity. These highly differentiated cells are vulnerable to an array of noxious stimuli that prevail in several glomerular diseases. Elevated circulating growth hormone (GH) levels are associated with podocyte injury and proteinuria in diabetes. However, the precise mechanism(s) by which excess GH elicits podocytopathy remains to be elucidated. Previous studies have shown that podocytes express GH receptor (GHR) and induce Notch signaling when exposed to GH. In the present study, we demonstrated that GH induces TGF-ß1 signaling and provokes cell cycle reentry of otherwise quiescent podocytes. Though differentiated podocytes reenter the cell cycle in response to GH and TGF-ß1, they cannot accomplish cytokinesis, despite karyokinesis. Owing to this aberrant cell cycle event, GH- or TGF-ß1-treated cells remain binucleated and undergo mitotic catastrophe. Importantly, inhibition of JAK2, TGFBR1 (TGF-ß receptor 1), or Notch prevented cell cycle reentry of podocytes and protected them from mitotic catastrophe associated with cell death. Inhibition of Notch activation prevents GH-dependent podocyte injury and proteinuria. Similarly, attenuation of GHR expression abated Notch activation in podocytes. Kidney biopsy sections from patients with diabetic nephropathy (DN) show activation of Notch signaling and binucleated podocytes. These data indicate that excess GH induced TGF-ß1-dependent Notch1 signaling contributes to the mitotic catastrophe of podocytes. This study highlights the role of aberrant GH signaling in podocytopathy and the potential application of TGF-ß1 or Notch inhibitors, as a therapeutic agent for DN.

Role of viral suppressors governing asymmetric synergism between tomato-infecting begomoviruses.

Mixed viral infections are common in fields and frequently exacerbate disease severity via synergistic interactions among individual viral genomic components leading to major crop loss. Two predominant species of tomato-infecting begomoviruses, Tomato leaf curl New Delhi virus (ToLCNDV) and Tomato leaf curl Gujarat virus (ToLCGuV), are known to cause severe leaf curl disease of tomato in India. Previously, we have demonstrated asymmetric synergism between these two distinct begomovirus species during mixed infection in solanaceous hosts. In the present study, we have identified the underlying proteins that positively regulate asymmetric synergism and their effect on plant defense machinery. During co-infection, the AC2 and AV2 of ToLCGuV enhanced ToLCNDV DNA accumulation in Nicotiana benthamiana as well as in their natural host, tomato. Furthermore, we found that AC2 and AV2 of ToLCNDV and AV2 of ToLCGuV play a critical role in suppression of post transcriptional gene silencing (PTGS) machinery. Taken together, AC2 and AV2 encoded proteins of ToLCGuV are the crucial viral factors promoting asymmetric synergism with ToLCNDV. KEY POINTS: • Begomoviral suppressors play vital roles in viral synergism. • AC2 and AV2 of ToLCGuV asymmetrically enhance ToLCNDV accumulation. • AC2 and AV2 of ToLCNDV and ToLCGuV AV2 are major PTGS suppressors.

Correlation between weather and COVID-19 pandemic in India: An empirical investigation.

This study is an attempt to find and analyze the correlation between Covid-19 pandemic and weather conditions in Indian context. Secondary data analysis of surveillance data of COVID-19 is taken from Wikipedia (updating information from World Health Organization) & statista.com and weather data through Power Data Access Viewer (DAV) (power.Iarc.nasa.gov) from NASA after mentioning latitude and longitude of India. The minimum temperature (°C) at 2 metre, maximum temperature (°C) at 2 metre, temperature (°C) at 2 metre and relative humidity (%) are taken as component of weather. To find the association, Spearman's rank correlation test was applied. The minimum, maximum temperature (°C) at 2 m, temperatures (°C) at 2 m and humidity at 2 m are significantly correlated with COVID-19 pandemic cases (r = 0.93, 0.94, 0.83, and 0.30) at 99% two-tailed significance level. The findings serve as an initial evidence to reduce the incidence rate of COVID-19 in India and useful in policy making.

Tobacco RNA-dependent RNA polymerase 1 affects the expression of defence-related genes in Nicotiana benthamiana upon Tomato leaf curl Gujarat virus infection.

MAIN CONCLUSION: RNA-dependent RNA polymerase 1 of Nicotiana tabacum modulates ToLCGV pathogenesis by influencing a number of defence-related genes in N. benthamiana plants. Key means of plants protecting themselves from the invading viruses is through RNA silencing. RNA-dependent RNA polymerase-1 (RDR1) is one of the crucial proteins of the RNA silencing pathway, which is induced after infection by viruses. RDR1 functions in the generation of small interfering RNAs (siRNAs) against the viral genome, thus it is antiviral in nature. Here, we used the transgenic Nicotiana benthamiana plant expressing N. tabacum NtRDR1 and observed reduced susceptibility towards Tomato leaf curl Gujarat virus (ToLCGV) infection compared to the wild-type N. benthamiana plants. To understand the reason for such reduced susceptibility, we prepared high-definition small RNA (sRNA) cDNA libraries from ToLCGV-infected wild-type N. benthamiana and NtRDR1 expressing N. benthamiana lines and carried out next-generation sequencing (NGS). We found that upon ToLCGV infection the majority of siRNAs generated from the host genome were of the 24 nucleotide (nt) class, while viral siRNAs (vsiRNAs) were of the 21-22-nt class, indicating that transcriptional gene silencing (TGS) is the major pathway for silencing of host genes while viral genes are silenced, predominantly, by post transcriptional gene silencing (PTGS) pathways. We estimated the changes in the expression of various defence-related genes, such as Constitutively Photomorphogenic-9 (COP9) signalosome (CSN) complex subunit-7, Pentatricopeptide repeat containing protein (PPRP), Laccase-3, Glutathione peroxidase-1 (GPX-1), Universal stress protein (USP) A-like protein, Heat shock transcription factor B4 (HSTF-B4), Auxin response factor-18 (ARF18), WRKY-6 and Short chain dehydrogenase reductase-3a. The differential expression of these genes might be linked with the enhanced tolerance of NtRDR1 N. benthamiana transgenic plants to ToLCGV. Our study suggests that reduced expression of subunit-7 of CSN complex and WRKY6, and increased expression of USPA-like protein might be linked with the reduced susceptibility of NtRDR1-transgenic N. benthamiana plants to ToLCGV.

Targeted sequencing of genes associated with the mismatch repair pathway in patients with endometrial cancer.

Germline variants inactivating the mismatch repair (MMR) genes MLH1, MSH2, MSH6 and PMS2 cause Lynch syndrome that implies an increased cancer risk, where colon and endometrial cancer are the most frequent. Identification of these pathogenic variants is important to identify endometrial cancer patients with inherited increased risk of new cancers, in order to offer them lifesaving surveillance. However, several other genes are also part of the MMR pathway. It is therefore relevant to search for variants in additional genes that may be associated with cancer risk by including all known genes involved in the MMR pathway. Next-generation sequencing was used to screen 22 genes involved in the MMR pathway in constitutional DNA extracted from full blood from 199 unselected endometrial cancer patients. Bioinformatic pipelines were developed for identification and functional annotation of variants, using several different software tools and custom programs. This facilitated identification of 22 exonic, 4 UTR and 9 intronic variants that could be classified according to pathogenicity. This study has identified several germline variants in genes of the MMR pathway that potentially may be associated with an increased risk for cancer, in particular endometrial cancer, and therefore are relevant for further investigation. We have also developed bioinformatics strategies to analyse targeted sequencing data, including low quality data and genomic regions outside of the protein coding exons of the relevant genes.

Activation of Notch1 signaling in podocytes by glucose-derived AGEs contributes to proteinuria.

INTRODUCTION: Advanced glycation end-products (AGEs) are implicated in the pathogenesis of diabetic nephropathy (DN). Previous studies have shown that AGEs contribute to glomerulosclerosis and proteinuria. Podocytes, terminally differentiated epithelial cells of the glomerulus and the critical component of the glomerular filtration barrier, express the receptor for AGEs (RAGE). Podocytes are susceptible to severe injury during DN. In this study, we investigated the mechanism by which AGEs contribute to podocyte injury. RESEARCH DESIGN AND METHODS: Glucose-derived AGEs were prepared in vitro. Reactivation of Notch signaling was examined in AGE-treated human podocytes (in vitro) and glomeruli from AGE-injected mice (in vivo) by quantitative reverse transcription-PCR, western blot analysis, ELISA and immunohistochemical staining. Further, the effects of AGEs on epithelial to mesenchymal transition (EMT) of podocytes and expression of fibrotic markers were evaluated. RESULTS: Using human podocytes and a mouse model, we demonstrated that AGEs activate Notch1 signaling in podocytes and provoke EMT. Inhibition of RAGE and Notch1 by FPS-ZM1 (N-Benzyl-4-chloro-N-cyclohexylbenzamide) and DAPT (N-[N-(3,5-Difluorophenacetyl)-L-alanyl]-S-phenyl glycine t-butylester), respectively, abrogates AGE-induced Notch activation and EMT. Inhibition of RAGE and Notch1 prevents AGE-induced glomerular fibrosis, thickening of the glomerular basement membrane, foot process effacement, and proteinuria. Furthermore, kidney biopsy sections from people with DN revealed the accumulation of AGEs in the glomerulus with elevated RAGE expression and activated Notch signaling. CONCLUSION: The data suggest that AGEs activate Notch signaling in the glomerular podocytes. Pharmacological inhibition of Notch signaling by DAPT ameliorates AGE-induced podocytopathy and fibrosis. Our observations suggest that AGE-induced Notch reactivation in mature podocytes could be a novel mechanism in glomerular disease and thus could represent a novel therapeutic target.

Conversion of asymptomatic infection to symptomatic visceral leishmaniasis: A study of possible immunological markers.

INTRODUCTION: Presence of asymptomatic individuals in endemic areas is common. The possible biomarkers in asymptomatic individuals once they get exposed to infection as well as following conversion to symptomatic disease are yet to be identified.We identified asymptomatic Visceral leishmaniasis (VL) infection amongst rK39+sorted direct agglutination test positive (DAT+) endemic healthy population and confirmed it by quantitative PCR(qPCR).The immunological determinants such as Adenosine deaminase (ADA), Interferon gamma (IFN-γ), Tumour Necrosis Factor alpha (TNF-α) and Interleukin 10 (IL-10)were examined to predict probable biomarkers for conversion to symptomatic VL. METHODS: Sample size was 5794 healthy individuals from VL endemic region. Antibody tests(DAT &rK39) were performed and later a qPCR assay was employed using kDNA specific primers and probes. Immunological biomarkers examined were ADA level by ADA-MTP kit and quantitative cytokines(IFN-γ, IL-10 and TNF-α) by ELISA. RESULTS: 120 asymptomatic individuals of 308 rK39 sero-positives were DAT positive comprising of 56 with previous history and 64 with no history of VL. RT-PCR confirmed asymptomatic VL in 42 sero-positives. These were followed up through repeated qPCR and evaluation of immunological determinants. We observed10 symptomatic cases converted from a total of 42 asymptomatic individuals identified at base-line. The level of ADA, IL-10 and IFN-γ remained consistently high in asymptomatic cases and amongst these, ADA and IL-10 but not IFN-γ remained higher at the development of clinical symptoms into active VL. On the contrary, there was no significant change in the mean concentration of TNF-α at both stages of the disease. DISCUSSION: We surmise from our data that considerable proportion of asymptomatic cases can be a reservoir and may play a crucial role in transmission of visceral leishmaniasis in endemic areas. The data also suggests that ADA and IL-10 can serve as a potential biomarker during the conversion of asymptomatic into symptomatic VL.

Nicotiana benthamiana phosphatidylinositol 4-kinase type II regulates chilli leaf curl virus pathogenesis.

Geminiviruses are single-stranded DNA viruses that can cause significant losses in economically important crops. In recent years, the role of different kinases in geminivirus pathogenesis has been emphasized. Although geminiviruses use several host kinases, the role of phosphatidylinositol 4-kinase (PI4K) remains obscure. We isolated and characterized phosphatidylinositol 4-kinase type II from Nicotiana benthamiana (NbPI4KII) which interacts with the replication initiator protein (Rep) of a geminivirus, chilli leaf curl virus (ChiLCV). NbPI4KII-mGFP was localized into cytoplasm, nucleus or both. NbPI4KII-mGFP was also found to be associated with the cytoplasmic endomembrane systems in the presence of ChiLCV. Furthermore, we demonstrated that Rep protein directly interacts with NbPI4KII protein and influenced nuclear occurrence of NbPI4KII. The results obtained in the present study revealed that NbPI4KII is a functional protein kinase lacking lipid kinase activity. Downregulation of NbPI4KII expression negatively affects ChiLCV pathogenesis in N. benthamiana. In summary, NbPI4KII is a susceptible factor, which is required by ChiLCV for pathogenesis.