Agriculture models for restoring degraded land to enhance CO2 biosequestration and carbon credits in the Vindhyan region of India.

The study's objective was to evaluate the status of converted degraded land into productive agricultural models by improving the physicochemical properties of the soil, soil organic matter (SOM), soil organic carbon (SOC) fractions (active and passive), and microbial biomass carbon (MBC), while also generating carbon (C) credit for additional farmers' income. Six models were analyzed, namely: (1) Arjun forest-based agroecosystems (AFBAE); (2) Lemon grass-based agroecosystems (LGBAE); (3) Legume-cereal-moong-based agroecosystems (LCMBAE); (4) Bael-black mustard-based agroecosystems (BMBAE); (5) Guava-wheat-based agroecosystems (GWBAE), and (6) Custard apple -lentil -based agroecosystems (CALBAE). These models were replicated three times in a randomized block design (RBD). Soil samples were collected from the study area at two depths (0-0.30 and 0.30-0.60 m). At a 0-0.30 m depth, the highest bulk density (ρb) of 1.50 Mg m-3 was observed in LCMBAE, while the lowest ρb of 1.43 Mg m-3 was recorded in BMBAE. The soil organic carbon (SOC) and SOC stock values exhibited a range of 4.2-7.7 g kg-1 and 19.0-33.4 Mg ha-1, respectively. In the AFBAE, the highest levels of 163.1 % MBC were found over LCMBAE. At a 0-0.30 m depth, the recalcitrant index (RI) and lability index (LI) ranged from 0.35-0.46 to 1.97-2.11, respectively. Additionally, the AFBAE exhibited the highest total biomass accumulation (39.23 Mg ha-1), carbon dioxide (CO2) biosequestration (287.9 Mg ha-1), and the total social cost of CO2 at US$ 277 ha-1. Furthermore, in the AFBAE, there was a 198.1 % increase in total C credit (US$ 161 ha-1) compared to LCMBAE (US$ 54 ha-1). However, at 0.30-0.60 m depths, GWBAE and CALBAE were statistically equivalent (p ≤ 0.05) in total C stocks. Principal component analysis (PCA) reveals that component-1 accounts for 77.4 % of the variability, while component-2 contributes 18.6 %. This article aimed to convert the degraded land into a sustainable agricultural module by increasing SOC and CO2 biosequestration and producing more C-credit, or climate currency, on underutilized land.

CD73-Dependent Adenosine Signaling through Adora2b Drives Immunosuppression in Ductal Pancreatic Cancer.

The microenvironment that surrounds pancreatic ductal adenocarcinoma (PDAC) is profoundly desmoplastic and immunosuppressive. Understanding triggers of immunosuppression during the process of pancreatic tumorigenesis would aid in establishing targets for effective prevention and therapy. Here, we interrogated differential molecular mechanisms dependent on cell of origin and subtype that promote immunosuppression during PDAC initiation and in established tumors. Transcriptomic analysis of cell-of-origin-dependent epithelial gene signatures revealed that Nt5e/CD73, a cell-surface enzyme required for extracellular adenosine generation, is one of the top 10% of genes overexpressed in murine tumors arising from the ductal pancreatic epithelium as opposed to those rising from acinar cells. These findings were confirmed by IHC and high-performance liquid chromatography. Analysis in human PDAC subtypes indicated that high Nt5e in murine ductal PDAC models overlaps with high NT5E in human PDAC squamous and basal subtypes, considered to have the highest immunosuppression and worst prognosis. Multiplex immunofluorescent analysis showed that activated CD8+ T cells in the PDAC tumor microenvironment express high levels of CD73, indicating an opportunity for immunotherapeutic targeting. Delivery of CD73 small-molecule inhibitors through various delivery routes reduced tumor development and growth in genetically engineered and syngeneic mouse models. In addition, the adenosine receptor Adora2b was a determinant of adenosine-mediated immunosuppression in PDAC. These findings highlight a molecular trigger of the immunosuppressive PDAC microenvironment elevated in the ductal cell of origin, linking biology with subtype classification, critical components for PDAC immunoprevention and personalized approaches for immunotherapeutic intervention. SIGNIFICANCE: Ductal-derived pancreatic tumors have elevated epithelial and CD8+GZM+ T-cell CD73 expression that confers sensitivity to small-molecule inhibition of CD73 or Adora2b to promote CD8+ T-cell-mediated tumor regression. See related commentary by DelGiorno, p. 977.

Assessment of optic disk by disk damage likelihood scale staging using slit-lamp biomicroscopy and optical coherence tomography in diagnosing primary open-angle glaucoma.

Purpose: The current study was aimed at assessment of optic disk by disk damage likelihood scale (DDLS) staging using slit-lamp biomicroscopy and optical coherence tomography (OCT) in diagnosing primary open-angle glaucoma (POAG) patients. Methods: This was a cross-sectional observational study of 106 POAG patients, which was conducted from April 2017 to April 2018. All patients underwent slit-lamp fundoscopy with a +78 D lens and high-definition (HD)-OCT, and the vertical cup disk ratios (VCDRs) were recorded. Disk size and neuroretinal rim assessment were done, and the disk was then staged using the recent version, which stages the optic nerve head (ONH) from 1 to 10 as read from the DDLS nomogram table. DDLS scores >5 indicate glaucomatous damage. Pearson coefficient was used to correlate the DDLS staging by slit-lamp biomicroscopy with best-corrected visual acuity (BCVA), intraocular pressure (IOP), disk size, and VCDR and VCDR, mean deviation, and DDLS staging by HD-OCT. Results: The mean age of the patients was 59.54 ± 6.61 years. The male: female ratio was 2:1. The mean IOP was 16.04 ± 1.97 mmHg, and BCVA was 0.72 ± 0.13 LogMAR units. The mean VCDR on 78 D slit-lamp biomicroscopy was 0.76 ± 0.09 (standard deviation [SD]) (range 0.1-0.77), whereas on HD-OCT, the mean VCDR was 0.81 ± 0.09 (SD) (range 0.07-0.81). The mean deviation on visual field testing in decibels was -14.43 ± 3.31 (SD). The correlation coefficient between DDLS staging by slit-lamp biomicroscopy and DDLS staging by HD-OCT parameters was r = 0.96. Conclusion: There is a positive correlation between the DDLS system of optic disk evaluation on slit-lamp biomicroscopy and most of the HD-OCT evaluation parameters.

Comparative transcriptome profiling of Polianthes tuberosa during a compatible interaction with root-knot nematode Meloidogyne incognita.

BACKGROUND: The root-knot nematode (RKN; Meloidogyne spp.) is the most destructive plant parasitic nematode known to date. RKN infections, especially those caused by Meloidogyne incognita, are one of the most serious diseases of tuberose. METHODS AND RESULTS: To investigate the molecular mechanism in the host-pathogen interactions, the Illumina sequencing platform was employed to generate comparative transcriptome profiles of uninfected and Meloidogyne incognita-infected tuberose plants, during early, mid, and late infection stage. A total of 7.5 GB (49 million reads) and 9.3 GB (61 million reads) of high-quality data was generated for the control and infected samples, respectively. These reads were combined and assembled using the Trinity assembly program which clustered them into 1,25,060 unigenes. A total of 85,360 validated CDS were obtained from the combined transcriptome whereas 6,795 CDS and 7,778 CDS were found in the data for the control and infected samples, respectively. Gene ontology terms were assigned to 958 and 1,310 CDSs from the control and infected data, respectively. The KAAS pathway analysis revealed that 1,248 CDS in the control sample and 1,482 CDS in the infected sample were enriched with KEGG pathways. The major proportions of CDS were annotated for carbohydrate metabolism, signal transduction and translation related pathways in control and infected samples. Of the 8,289 CDS commonly expressed between the control and infected plants, 256 were significantly upregulated and 129 were significantly downregulated in the infected plants. CONCLUSIONS: Collectively, our results provide a comprehensive gene expression changes in tuberose during its association with RKNs and point to candidate genes that are involved in nematode stress signaling for further investigation. This is the first report addressing genes associated with M. incognita-tuberose interaction and the results have important implications for further characterization of RKN resistance genes in tuberose.

Germi-X herbal-based spray disinfects smartphone surfaces: implication on fomite-mediated infection spread.

Inanimate objects/surfaces become fomites upon contacting infectious agents such as disease-causing bacteria, fungi and viruses. Smartphones are one of the most prominent among these fomites. COVID-19 pandemic has raised the awareness on mobile sanitization, as an active measure to curb fomite-mediated viral transmission. Available mobile sanitizers and ultraviolet (UV) ray mediated mobile sanitization have their own sets of pros and cons, often being less user-friendly. This study explored the germicidal efficacy of an herbal-based sanitizer, Germi-X spray, on hands and mobiles, through microbiological techniques of micro-broth dilution and Kirby-Bauer disc diffusion assay, thumb print assay and swab test. Notably, Germi-X spray was found to be 6-67% more effective against surface pathogens, like, Staphylococcus epidermidis, Staphylococcus aureus, Pseudomonas fluorescens and Pseudomonas aeruginosa, as compared to a very popular product in the Indian market, which was taken as a control for this study. The observed anti-bacterial activity of the spray from disc-diffusion assay suggests its greater surface retentivity as compared to the control. Germicidal potency of Germi-X spray, when used to sanitize hands, was found to be greater than 80%. There was ~ 17-fold reduction in microbial counts after sanitizing smartphones with Germi-X spray. The novelty of this study lies in providing experimental evidence for this herbal-based surface sanitizer in efficiently disinfecting one of the super contaminated fomite, the smartphones. In conclusion, having an herbal base with a high germicidal efficacy against surface pathogens, together with longer surface retention, Germi-X spray appears to be an eco-friendly and cost-effective sanitizer for the surfaces of electronic gadgets like smartphones.

Pre-treatment hematological parameters as a cost effective predictive marker for response to concurrent chemo radiation in locally advanced cervical cancer.

INTRODUCTION: Locally advanced cervical cancer is still a major cause of mortality in developing countries. Recently, personalized medicine has changed the treatment paradigm for many solid cancers but no robust biomarkers has yet been validated for predicting response to chemo radiation in cervical cancer patients. AIM: To assess the role of hematological parameters as a cost-effective predictive marker of response to concurrent chemo radiation in cervical cancer patients. MATERIALS AND METHOD: This is a retrospective analysis of 90 cervical cancer patients treated with concurrent chemo radiation in a tertiary cancer center. Clinical details of the patients were extracted from the case records. For end point evaluation, the pre-treatment levels of hemoglobin, neutrophil, lymphocyte, platelet, platelet lymphocyte ratio (PLR) and neutrophil lymphocyte ratio (NLR) were compared and statistically analyzed between responders and non-responders. The optimal cutoff values of hematological parameters were estimated by the receiver operating characteristics (ROC) curve. RESULT: Out of 90 patients, 60 (66.66%) were complete responders and remaining 30 (33.33%) were non-responders. The mean value of platelet, NLR, and PLR was significantly higher in the non-responder group. ROC curve analysis showed the optimal cut-off value of pre-treatment Hb, PLT, NLR and PLR to be 11 gm/dl, 3, 177 × 109/L, and 70 respectively. CONCLUSION: Our study suggests that simple hematological markers like NLR, PLT count and PLR could be used as a cost effective pretreatment predictive marker for response to chemo radiation in cervical cancer patients.

Murine Models for Lineage Tracing Cancer Initiating Cells.

The process of cellular transformation involves acquisition of genetic mutations that disrupt parenchymal organization and promote unrestrained epithelial cell proliferation. In the context of transformation, recent advances using genetic lineage tracing have enabled scientists to study the behavior of neoplastic cells in their native habitat. Employing lineage tracing to understand mechanisms of transformation, including cell type-specific responses to inflammation, is an empirical method to use in the field of cancer prevention. Identifying new approaches to prevent cancer formation can be directly studied by indelibly labeling cells to trace their fate and understand at a molecular level how they respond to novel methods for testing cancer preventing agents.

Anu taila, an herbal nasal drop, suppresses mucormycosis by regulating host TNF-α response and fungal ergosterol biosynthesis.

AIM: The intractable, mucormycosis, caused by Mucorales primarily targets immunocompromised individuals. The first-line therapy, intravenous liposomal amphotericin B and surgical debridement of necrotic tissue, is contraindicative in individuals with compromised kidneys. This invokes a pressing need to identify safer treatment options. METHODS AND RESULTS: The antifungal effect of the classical nasal drop, Anu taila, against Mucor spp. was investigated through microbiological, cytological, analytical chemical (HPLC and GS-MS/MS) and scanning electron microscopic (SEM) approaches. Anu taila-pretreated spores germinated late, resulting in reduced infectivity, observed as milder monocytic immune response. Conversely, Anu taila-pretreated human THP-1 cells exhibited an improved immune response against Mucor spores, through TNF-α. Repeated Anu taila application rapidly abolished fungal microarchitectures than amphotericin B, evident from swift replacement of hyphae, sporangiophores and sporangia with fused biomass, in the SEM images. HPLC analysis showed that Anu taila treatment significantly reduced overall ergosterol content in Mucor biomass. Anu taila also downregulated sterol-C5-desaturase-coding ERG3 gene, crucial for ergosterol biosynthesis and resultant structural integrity, in Mucor spp. CONCLUSION: Taken together, Anu taila was found effective against Mucor spp., with both prophylactic and curative implications, which is attributable to the phytochemical composition of this classical nasal drop. SIGNIFICANCE AND IMPACT STATEMENT: The potential remedial effects of a classical nasal drop against an obdurate and challenging fungal infection are identified.

Vishaghn Dhoop, Nano-Scale Particles with Detoxifying Medicinal Fume, Exhibits Robust Anti-Microbial Activities: Implications of Disinfection Potentials of a Traditional Ayurvedic Air Sterilization Technique.

The rapidly increasing global burden of healthcare associated infections (HAI) is resulting in proportionate increase in chemical disinfection in healthcare settings, adding an extra burden of environmental toxicity. Therefore, alternative disinfection techniques with less or no adverse side-effects need to be explored. In this regard, ayurvedic 'dhoopan' technique involving slow combustion of medicinal herbs, minerals and animal products hold great promise. In this study, dhoopan of a traditionally defined ayurvedic medicinal mix, 'Vishaghn Dhoop' (VD) has been assessed for its anti-microbial potentials against both Gram-positive and negative pathogenic bacteria, Mycobacterium and pathogenic fungus, Candida albicans. Fume generated from slow combustion of VD was subjected to physico-chemical characterization and was assessed for anti-microbial effects. VD fume contained particles of 354 ± 84 nm size, laden with anti-microbial metabolites. On agar plates, VD fumigation reduced bacterial growth by 13 - 38%. Liquid culture aeration with VD fume inhibited bacterial growth by 50 - 85%, and fungal growth by 80%. In real life settings (in vivo), un-sanitized rooms fumigated with VD fumes for 30 min reduced the environmental microbial loads by 10 folds. In addition, the safety of VD fumigation was evaluated through in vitro cytotoxicity assay on human lung epithelial (A549) cells. Cells exposed to media-collected VD fumes for 24 h exhibited normal cyto-safety profile. Collectively, these observations provide scientific evidence in support of a traditional technique of disinfection, which can be fine-tuned to have implications in clinical, healthcare and food industry where, disinfection is a prime requirement.

Kras mutation rate precisely orchestrates ductal derived pancreatic intraepithelial neoplasia and pancreatic cancer.

Pancreatic ductal adenocarcinoma (PDAC) is the third leading cause of cancer-related death in the United States. Despite the high prevalence of Kras mutations in pancreatic cancer patients, murine models expressing the oncogenic mutant Kras (Krasmut) in mature pancreatic cells develop PDAC at a low frequency. Independent of cell of origin, a second genetic hit (loss of tumor suppressor TP53 or PTEN) is important for development of PDAC in mice. We hypothesized ectopic expression and elevated levels of oncogenic mutant Kras would promote PanIN arising in pancreatic ducts. To test our hypothesis, the significance of elevating levels of K-Ras and Ras activity has been explored by expression of a CAG driven LGSL-KrasG12V allele (cKras) in pancreatic ducts, which promotes ectopic Kras expression. We predicted expression of cKras in pancreatic ducts would generate neoplasia and PDAC. To test our hypothesis, we employed tamoxifen dependent CreERT2 mediated recombination. Hnf1b:CreERT2;KrasG12V (cKrasHnf1b/+) mice received 1 (Low), 5 (Mod) or 10 (High) mg per 20 g body weight to recombine cKras in low (cKrasLow), moderate (cKrasMod), and high (cKrasHigh) percentages of pancreatic ducts. Our histologic analysis revealed poorly differentiated aggressive tumors in cKrasHigh mice. cKrasMod mice had grades of Pancreatic Intraepithelial Neoplasia (PanIN), recapitulating early and advanced PanIN observed in human PDAC. Proteomics analysis revealed significant differences in PTEN/AKT and MAPK pathways between wild type, cKrasLow, cKrasMod, and cKrasHigh mice. In conclusion, in this study, we provide evidence that ectopic expression of oncogenic mutant K-Ras in pancreatic ducts generates early and late PanIN as well as PDAC. This Ras rheostat model provides evidence that AKT signaling is an important early driver of invasive ductal derived PDAC.

GermiX: A skin friendly hand sanitizer with prolonged effectivity against pathogenic bacteria.

COVID-19 pandemic has almost made hand sanitization a ritual resulting in a steep increase in the frequency of hand sanitization and an unprecedented surge in demand for hand sanitizers. In fact, several governments had to ration hand sanitizers in the retail outlets and over the counter chemist shops. Additionally, Indian government has put a cap on the prices of hand sanitizers. Currently, large sections of global and Indian population are grappling under financial crises. Therefore, mandatory hand sanitization has made an unwelcoming, yet unavoidable addition to the already-hard-to-maintain-grocery-list. Here, we have compared the anti-microbial efficacy of Patanjali Hand Sanitizer (PHS), developed and marketed by Patanjali Ayurved Ltd. (an India-based food and herbal medicine company) with one of the topmost hand sanitizers currently used under clinical set-ups. PHS has anti-microbial efficacy comparable to that of the standard hand sanitizer. Besides, disc diffusion and time-dependent thumb print assays showed that PHS has longer retentivity on the applied surfaces, suggesting lesser consumption of the sanitizer and concomitant relaxation on the monthly grocery budget. Observed anti-bacterial potency of PHS is attributed to the disruption of bacterial cell membrane, as employed by alcohol-based hand sanitizers. A rough estimation revealed that PHS is ~ 4.3 times cost effective than the standard hand sanitizer used as the positive control in this study. Taken together, PHS is a suitable alternative for existing hand sanitizers available in the market that can relax the demand-supply strain and soften significantly the burden of monthly expenditure on hand sanitizers.

Mst1/2 kinases restrain transformation in a novel transgenic model of Ras driven non-small cell lung cancer.

Non-small cell lung cancer remains a highly lethal malignancy. Using the tamoxifen inducible Hnf1b:CreERT2 (H) transgenic mouse crossed to the LsL-KrasG12D (K) transgenic mouse, we recently discovered that an Hnf1b positive cell type in the lung is sensitive to adenoma formation when expressing a mutant KrasG12D allele. In these mice, we observe adenoma formation over a time frame of three to six months. To study specificity of the inducible Hnf1b:CreERT2 in the lung, we employed lineage tracing using an mTmG (G) reporter allele. This technique revealed recombined, GFP+ cells were predominantly SPC+. We further employed this technique in HKG mice to determine Hnf1b+ cells give rise to adenomas that express SPC and TTF1. Review of murine lung tissue confirmed a diagnosis of adenoma and early adenocarcinoma, a pathologic subtype of non-small cell lung cancer. Our expanded mouse model revealed loss of Mst1/2 promotes aggressive lung adenocarcinoma and large-scale proteomic analysis revealed upregulation of PKM2 in the lungs of mice with genetic deletion of Mst1/2. PKM2 is a known metabolic regulator in proliferating cells and cancer. Using a human lung adenocarcinoma cell line, we show pharmacologic inhibition of Mst1/2 increases the abundance of PKM2, indicating genetic loss or pharmacologic inhibition of Mst1/2 directly modulates the abundance of PKM2. In conclusion, here we report a novel model of non-small cell lung cancer driven by a mutation in Kras and deletion of Mst1/2 kinases. Tumor development is restricted to a subset of alveolar type II cells expressing Hnf1b. Our data show loss of Mst1/2 regulates levels of a potent metabolic regulator, PKM2.

Mutant p53R175H promotes cancer initiation in the pancreas by stabilizing HSP70.

Pancreatic cancer remains a highly lethal malignancy. We have recently shown that simultaneous expression of Kras and mutant Tp53R175H promotes invasive ductal adenocarcinoma from pancreatic ductal cells. We hypothesized specific mutations in TP53 have divergent mechanisms of transforming ductal cells. In order to understand the role of mutant TP53 in transforming pancreatic ductal cells, we used a lentiviral system to express mutant TP53R175H and TP53R273H, two of the most frequently mutated TP53 alleles in pancreatic cancer patients, in immortalized, but not transformed, pancreatic ductal epithelial cells carrying a KRAS mutation (HPNE:KRASG12D). Mutant TP53 expression enhanced colony formation and an RPPA assay results revealed TP53R175H uniquely induced HSP70 expression in HPNE:KRASG12D cells. In the context of TP53R175H expression; we observed nuclear localization of HSP70. We performed immunoprecipitation experiments to show mutant p53R175H binds to HSP70. We also provide evidence mutant p53R175H is important for HSP70 stability and, more importantly, HSP70 is required for mutant p53 stability. These data are critical in the context of events leading to cellular transformation in the pancreas.

Identification of putative flowering genes and transcription factors from flower de novo transcriptome dataset of tuberose (Polianthes tuberosa L.).

Polianthes tuberosa is commercially popular because of their economic importance in floriculture for cut and loose flowers and in perfume industry because of the unique fragrance. Despite its commercial importance, no ready-to-use transcript sequence information is available in the public database. We have sequenced the RNA obtained from tuberose flowers using the Illumina HiSeq. 2000 platform and have carried out a de novo analysis of the transcriptome data. The de novo assembly generated 11,100 transcripts. These transcripts represent a total of 7876 unigenes that were considered for downstream analysis. These 7876 unigenes, which was further annotated using blast2go and KEGG pathways, were also assigned. Tuberose transcripts were also assigned to metabolic pathways using the Kyoto Encyclopedia of Genes and Genomes database to determine their biochemical functions. 4591 of the tuberose transcripts matched to genes in KEGG pathways and 66 transcripts were mapped to the Flavonoid biosynthesis pathway. 21 flowering genes have been identified in this tuberose transcriptome. Transcription factor analysis helped in the identification of a large number of transcripts similar to key genes in the flowering regulation network of Arabidopsis thaliana. Among the transcription factors identified "NAC" which is associated with plant stress response represented the most abundant category followed by APETALA2 (AP2)/ethylene-responsive element binding proteins (EREBPs) which plays various role in floral organ identity and respond to different biotic and abiotic stress.

Expression of estrogen receptor beta isoforms in pancreatic adenocarcinoma.

Limited studies have shown that some patients with pancreatic adenocarcinoma (PAC) may benefit from treatment with tamoxifen. PAC has been shown to be largely negative for estrogen receptor alpha (ER-alpha). The aim of this pilot study was to investigate ER-beta expression in human PAC. Sections of tissue microarray with 18 evaluable cases of human PAC were stained by immunohistochemistry (IHC) for ER-beta1, ER-beta2, ER-beta5, and Cyclin A. The levels of ER-beta isoform expression and the S-phase fraction (SPF) were determined using quantitative digital image analysis. Higher mean and median ER-beta2 levels correlated with male sex (p = 0.057 and p = 0.035, respectively), older age (p = 0.005 and p = 0.006, respectively), and lower pT stage (p = 0.008 and p = 0.009). Mean and median ER-beta5 levels correlated negatively with SPF (p = 0.021 and p = 0.047, respectively). Mean ER-beta1 expression did not correlate with any of the above mentioned clinicopathologic factors. The findings in this pilot study, although should be considered preliminary, suggest that some ER-beta isoforms may play a role in the biology of PAC. Additional larger studies are needed to confirm our findings, and to determine whether ER-beta may be considered for future targeted therapy.

Association of Glycated Haemoglobin and Serum Apolipoproteins with Diabetic Retinopathy: An Indian Overview.

INTRODUCTION: India is presently facing an epidemic of diabetes mellitus and the risks of chronic complications from the disease are associated with the duration of the disease as well as the degree of hyperglycaemia. Diabetic retinopathy is a known microvascular complication of diabetes mellitus and is the most common cause of blindness in the western countries.Apolipoproteins are the protein component of lipoproteins. Apart from acting as structural proteins, they also act as cofactors to various enzymes. AIM: To measure the levels of serum apolipoproteins and glycated haemoglobin in cases of diabetic retinopathy and to assess their association with the stages of diabetic retinopathy. MATERIAL AND METHODS: The 135 diabetic cases [with (110) and without (125) retinopathy] attending the Ophthalmology OPD of this tertiary care hospital were included in the present study. Following retinoscopy, the patients were classified as Non-Proliferative Diabetic Retinopathy (NPDR) (n=75) and Proliferative Diabetic Retinopathy (PDR) (n=35). The controls (n=100) were age and sex matched patients who did not have diabetes. The cases and controls were assessed for HbA1c, total cholesterol, triglycerides, HDL cholesterol, Apo A-I and Apo B-100. RESULTS: The HbA1c was found to be higher in diabetics without retinopathy (7.02%) as compared to controls (5.58%) (p<0.05) and the highest value was seen in the mild NPDR group (8.82%). The mean value of Apo A-I was found to be lowest in the diabetics without retinopathy at 88 mg/dl and the highest in severe NPDR at 167 mg/dL. The mean value of Apo B-100 was found to be highest in severe NPDR at 114 mg/dL. The mean value of HDL cholesterol was lowest in moderate NPDR at 36.6 mg/dl. Total cholesterol was highest in severe NPDR at 280.88mg/dl while triglyceride was highest in severe NPDR at 286.4mg/dl. CONCLUSION: In our study, the level of HbA1c was found to range from 5.58% in non-diabetic to 8.82% in mild NPDR. There was a clear association between Apo B-100 and total cholesterol, triglycerides with the highest value of each parameter seen in the severe NPDR group. There was a discordance noted in the levels of HDL and Apo A-I in various groups. Apo B-100 values may be of value in prognosis of diabetic retinopathy as higher values may result in progression of the disease. Further studies involving Lp(a) and homocysteine may be required in cases of diabetic retinopathy.

An LC-MS/MS assay for the quantitative determination of 2-pyridyl acetic acid, a major metabolite and key surrogate for betahistine, using low-volume human K2 EDTA plasma.

Betahistine is widely used for the treatment of vertigo. Owing to first-pass metabolism, 2-pyridyl acetic acid (2PAA, major metabolite of betahistine) was considered as surrogate for quantitation. A specific and sensitive LC-MS/MS method was developed and validated for quantitation of 2PAA using turbo-ion spray in a positive ion mode. A solid-phase extraction was employed for the extraction of 2PAA and 2PAA d6 (IS) from human plasma. Chromatographic separation of analytes was achieved using an ACE CN, 5 µm (50 × 4.6 mm) column with a gradient mobile phase comprising acetonitrile-methanol (90:10% v/v) and 0.7% v/v formic acid in 0.5 mm ammonium trifluoroacetate in purified water (100% v/v). The retention times of 1.15 and 1.17 min for 2PAA and internal standard, respectively, were achieved. Quantitation of 2PAA and internal standard was achieved by monitoring multiple reaction monitoring transition pairs (m/z 138.1 to m/z 92.0 and m/z 142.1 to m/z 96.1, respectively). The developed method was validated for various parameters. The calibration curves of 2PAA showed linearity from 5.0 to 1500 ng/mL, with a lower limit of quantitation of 5.0 ng/mL. The bias and precision for inter- and intra-batch assays were <10%. The developed method was used to support clinical sample analysis.

Quantitative determination of saroglitazar, a predominantly PPAR alpha agonist, in human plasma by a LC-MS/MS method utilizing electrospray ionization in a positive mode.

A sensitive LC-MS/MS method was developed and validated for quantitation of saroglitazar using turboion spray interface with positive ion mode. A liquid-liquid extraction, with a mixture of dichloromethane and diethyl ether, was employed for the extraction of saroglitazar and glimepiride (IS) from human plasma. The chromatographic separation was achieved using an ACE-5, C18 (4.6 × 100 mm) column with a gradient mobile phase comprising acetonitrile and ammonium acetate buffer with trifluoracetic acid in purified water. Both analytes were separated within 10 min with retention times of 4.52 and 2.57 min for saroglitazar and IS, respectively. Saroglitazar quantitation was achieved by the summation of two MRM transition pairs (m/z 440.2 to m/z 366.0 and m/z 440.2 to m/z 183.1), while that of IS was achieved using transition pair m/z 491.3 to m/z 352.0. The calibration standards of saroglitazar showed linearity from 0.2 to 500 ng/mL, with a lower limit of quantitation of 0.2 ng/mL. The biases for inter- and intra-batch assays were -7.51-1.15% and -11.21 to -3.25%, respectively, while the corresponding precisions were 5.04-8.06% and 1.53-7.68%, respectively. The developed method was used to monitor the plasma concentrations of saroglitazar in clinical samples.

By moonlighting in the nucleus, villin regulates epithelial plasticity.

Villin is a tissue-specific, actin-binding protein involved in the assembly and maintenance of microvilli in polarized epithelial cells. Conversely, villin is also linked with the loss of epithelial polarity and gain of the mesenchymal phenotype in migrating, invasive cells. In this study, we describe for the first time how villin can switch between these disparate functions to change tissue architecture by moonlighting in the nucleus. Our study reveals that the moonlighting function of villin in the nucleus may play an important role in tissue homeostasis and disease. Villin accumulates in the nucleus during wound repair, and altering the cellular microenvironment by inducing hypoxia increases the nuclear accumulation of villin. Nuclear villin is also associated with mouse models of tumorigenesis, and a systematic analysis of a large cohort of colorectal cancer specimens confirmed the nuclear distribution of villin in a subset of tumors. Our study demonstrates that nuclear villin regulates epithelial-mesenchymal transition (EMT). Altering the nuclear localization of villin affects the expression and activity of Slug, a key transcriptional regulator of EMT. In addition, we find that villin directly interacts with a transcriptional corepressor and ligand of the Slug promoter, ZBRK1. The outcome of this study underscores the role of nuclear villin and its binding partner ZBRK1 in the regulation of EMT and as potential new therapeutic targets to inhibit tumorigenesis.