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Introduction: Chronic diseases, such as type 2 diabetes mellitus and hypertension, are often associated with psychiatric comorbidities such as anxiety, depression, and somatization. Approximately, one-fourth of the adults were diagnosed with hypertension, and the proportion will reach about one-third by 2025. The prevalence of hypertension throughout India is 29.8% and the burden of hypertension in a rural area of Tamil Nadu is 25.2%. The compliance of drug intake depends on the mental health of the patient and this study intends to take care of patients with chronic illnesses. This study is designed to assess the mental health of hypertensive patients and its association with their blood pressure. Materials and Methods: A cross-sectional descriptive study was conducted among hypertensive patients in the field practice of a tertiary care medical college in the Kancheepuram district for a period of 3 months using a semi-structured validated schedule after obtaining the informed consent. The depression anxiety stress scale (DASS) scale was used to assess the mental health of hypertensive patients. Data were analyzed using the Statistical Package for Social Sciences (SPSS) version 23.0. Results: Most hypertensive patients were found in the age group of 40 to 60 years. About 53.4% of hypertensive patients with normal blood pressure were suffering from depression. In addition, about 44.6% of pre-hypertensive patients and 44.6% of stage II hypertensive patients were found to have severe depression. Conclusion: Overall, pre-hypertensive patients were suffering from depression when compared to other hypertensive patients. Family history and tobacco and alcohol intake were other factors associated with depression in hypertensive patients. [ABSTRACT FROM AUTHOR]

Store-operated calcium entry (SOCE) is essential for cellular signaling. Earlier studies of the pyrazole derivative BTP2, an efficient inhibitor SOCE, identified that SOCE blockade suppresses proinflammatory gene expression. The impact of SOCE blockade on gene expression at the whole transcriptome level, however, is unknown. To fill this gap, we performed RNA sequencing (RNA-seq) and investigated at the whole transcriptome level the effect of BTP2 on gene expression in human peripheral blood mononuclear cells signaled with phytohemagglutinin. Our global gene expression analysis identified that SOCE blockade spares activation-induced expression of anti-inflammatory genes (e.g., IL10, TGFB1, FOXP3, and CTLA4) whereas the induced expression of proinflammatory genes such as IFNG and cytopathic genes such as GZMB are inhibited. We validated the differential expression of immunoregulatory genes identified by RNA-seq using preamplification-enhanced RT-qPCR assays. Because IL-2/IL2RA interaction is essential for T cell clonal expansion, we investigated and confirmed that BTP2 inhibits IL2RA expression at the protein level using multiparameter flow cytometry. Our elucidation that SOCE blockade spares activation-induced expression of anti-inflammatory genes while blocking pro-inflammatory gene expression suggests that SOCE blockers may represent a novel class of immunoregulatory drugs of value for treating autoimmune disease states and organ transplantation., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 American Society for Histocompatibility and Immunogenetics. Published by Elsevier Inc. All rights reserved.)

Introduction: Acute rejection (AR) undermines the life-extending benefits of kidney transplantation and is diagnosed using the invasive biopsy procedure. T cell-mediated rejection (TCMR), antibody-mediated rejection (ABMR), or concurrent TCMR + ABMR (Mixed Rejection [MR]) are the three major types of AR. Development of noninvasive biomarkers diagnostic of AR due to any of the three types is a useful addition to the diagnostic armamentarium., Methods: We developed customized RT-qPCR assays and measured urinary cell mRNA copy numbers in 145 biopsy-matched urine samples from 126 kidney allograft recipients. We determined whether the urinary cell three-gene signature diagnostic of TCMR (Suthanthiran et al., 2013) discriminates patients with no rejection biopsies (NR, n = 50) from those with ABMR (n = 28) or MR (n = 20) biopsies., Results: The urinary cell three-gene signature discriminated all three types of rejection biopsies from NR biopsies (P < 0.0001, One-way ANOVA). Dunnett's multiple comparisons test yielded P < 0.0001 for NR vs. TCMR; P < 0.001 for NR vs. ABMR; and P < 0.0001 for NR vs. MR. By bootstrap resampling, optimism-corrected area under the receiver operating characteristic curve (AUC) was 0.749 (bias-corrected 95% confidence interval [CI], 0.638 to 0.840) for NR vs. TCMR (P < 0.0001); 0.780 (95% CI, 0.656 to 0.878) for NR vs. ABMR (P < 0.0001); and 0.857 (95% CI, 0.727 to 0.947) for NR vs. MR (P < 0.0001). All three rejection categories were distinguished from NR biopsies with similar accuracy (all AUC comparisons P > 0.05)., Conclusion: The urinary cell three-gene signature score discriminates AR due to TCMR, ABMR or MR from NR biopsies in human kidney allograft recipients., Competing Interests: Declaration of competing interest M. Suthanthiran has a Consultancy Agreement with CareDx, Inc. Brisbane, CA. D.M. Dadhania D.M.D. has participated in industry sponsored studies organized by AlloVir Inc., CSL Behring, and Memo Therapeutics AG and served as consultant for CareDx, Inc. The other authors of this manuscript declare no conflicts of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Purpose: Estimation of multi-compartment intravoxel 'flow' in fD in ml/100g/min with multi-b-value diffusion weighted imaging and a multi-Gaussian model in the kidneys., Theory and Methods: A multi-Gaussian model of intravoxel flow using water transport time to quantify f D (ml/100g/min) is presented and simulated. Multi-compartment anisotropic DWI signal is simulated with Rician noise and SNR=50 and analyzed with a rigid bi-exponential, a rigid tri-exponential and diffusion spectrum imaging model of intravoxel incoherent motion (spectral diffusion) to study extraction of multi-compartment flow. The regularization parameter for spectral diffusion is varied to study the impact on the resulting spectrum and computation speed. The application is demonstrated in a two-center study of 54 kidney allografts with 9 b-value advanced DWI that were split by function (CKD-EPI 2021 eGFR<45ml/min/1.73m 2 ) and fibrosis (Banff 2017 interstitial fibrosis and tubular atrophy score 0-6) to demonstrate multi-compartment flow of various kidney pathologies., Results: Simulation of anisotropic multi-compartment flow from spectral diffusion demonstrated strong correlation to truth for both three-compartment anisotropic diffusion ( y = 1.08 x + 0.1 , R 2 = 0.71 ) and two-compartment anisotropic diffusion ( y = 0.91 + 0.6 , R 2 = 0.74 ), outperforming rigid models in cases of variable compartment number. Use of a fixed regularization parameter set to λ = 0.1 increased computation up to 208-fold and agreed with voxel-wise cross-validated regularization (concordance correlation coefficient=0.99). Spectral diffusion of renal allografts showed decreasing trend of tubular and vascular flow with higher levels of fibrosis, and significant increase in tissue parenchyma flow (f-stat=3.86, p=0.02). Tubular f D was significantly decreased in allografts with impaired function (eGFR<45ml/min/1.73m 2 )(Mann-Whitney U t-stat=-2.14, p=0.04)., Conclusions: Quantitative multi-compartment intravoxel 'flow' can be estimated in ml/100g/min with f D from multi-Gaussian diffusion with water transport time, even with moderate anisotropy such as in kidneys. The use of spectral diffusion with a multi-Gaussian model and a fixed regularization parameter is particularly promising in organs such as the kidney with variable numbers of physiologic compartments.

Intracellular Mg 2+ ( i Mg 2+ ) is bound with phosphometabolites, nucleic acids, and proteins in eukaryotes. Little is known about the intracellular compartmentalization and molecular details of Mg 2+ transport into/from cellular organelles such as the endoplasmic reticulum (ER). We found that the ER is a major i Mg 2+ compartment refilled by a largely uncharacterized ER-localized protein, TMEM94. Conventional and AlphaFold2 predictions suggest that ERMA (TMEM94) is a multi-pass transmembrane protein with large cytosolic headpiece actuator, nucleotide, and phosphorylation domains, analogous to P-type ATPases. However, ERMA uniquely combines a P-type ATPase domain and a GMN motif for ER Mg 2+ uptake. Experiments reveal that a tyrosine residue is crucial for Mg 2+ binding and activity in a mechanism conserved in both prokaryotic (mgtB and mgtA) and eukaryotic Mg 2+ ATPases. Cardiac dysfunction by haploinsufficiency, abnormal Ca 2+ cycling in mouse Erma +/- cardiomyocytes, and ERMA mRNA silencing in human iPSC-cardiomyocytes collectively define ERMA as an essential component of ER Mg 2+ uptake in eukaryotes., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Background: Delineation of T-cell genes, gene sets, pathways, and T-cell subtypes associated with acute T cell-mediated rejection (TCMR) may improve its management., Methods: We performed bulk RNA-sequencing of 34 kidney allograft biopsies (16 Banff TCMR and 18 no rejection [NR] biopsies) from 34 adult recipients of human kidneys. Computational analysis was performed to determine the differential intragraft expression of T-cell genes at the level of single-gene, gene set, and pathways., Results: T-cell signaling pathway gene sets for plenary T-cell activation were overrepresented in TCMR biopsies compared with NR biopsies. Heightened expression of T-cell signaling genes was validated using external TCMR biopsies. Pro- and anti-inflammatory immune gene sets were enriched, and metabolism gene sets were depleted in TCMR biopsies compared with NR biopsies. Gene signatures of regulatory T cells, Th1 cells, Th2 cells, Th17 cells, T follicular helper cells, CD4 tissue-resident memory T cells, and CD8 tissue-resident memory T cells were enriched in TCMR biopsies compared with NR biopsies. T-cell exhaustion and anergy were also molecular attributes of TCMR. Gene sets associated with antigen processing and presentation, and leukocyte transendothelial migration were overexpressed in TCMR biopsies compared with NR biopsies. Cellular deconvolution of graft infiltrating cells by gene expression patterns identified CD8 T cell to be the most abundant T-cell subtype infiltrating the allograft during TCMR., Conclusions: Our delineation of intragraft T-cell gene expression patterns, in addition to yielding new biological insights, may help prioritize T-cell genes and T-cell subtypes for therapeutic targeting., Competing Interests: The authors declare no conflicts of interest., (Copyright © 2024 Wolters Kluwer Health, Inc. All rights reserved.)

Competing Interests: The authors declare no conflicts of interest.

Natural killer (NK) cells mediate spontaneous cell-mediated cytotoxicity and antibody-dependent cell-mediated cytotoxicity. This dual functionality could enable their participation in chronic active antibody-mediated rejection (CA-ABMR). Earlier microarray profiling studies have not subcategorized antibody-mediated rejection into CA-ABMR and active-ABMR, and the gene expression pattern of CA-ABMR has not been compared with that of T cell-mediated rejection (TCMR). To fill these gaps, we RNA sequenced human kidney allograft biopsies categorized as CA-ABMR, active-ABMR, TCMR, or No Rejection (NR). Among the 15,910 genes identified in the biopsies, 60, 114, and 231 genes were uniquely overexpressed in CA-ABMR, TCMR, and active-ABMR, respectively; compared to NR, 50 genes were shared between CA-ABMR and active-ABMR, and 164 genes between CA-ABMR and TCMR. The overexpressed genes were annotated to NK cells and T cells in CA-ABMR and TCMR, and to neutrophils and monocytes in active-ABMR. The NK cell cytotoxicity and allograft rejection pathways were enriched in CA-ABMR. Genes encoding perforin, granzymes, and death receptor were overexpressed in CA-ABMR versus active-ABMR but not compared to TCMR. NK cell cytotoxicity pathway gene set variation analysis score was higher in CA-ABMR compared to active-ABMR but not in TCMR. Principal component analysis of the deconvolved immune cellular transcriptomes separated CA-ABMR and TCMR from active-ABMR and NR. Immunohistochemistry of kidney allograft biopsies validated a higher proportion of CD56 + NK cells in CA-ABMR than in active-ABMR. Thus, CA-ABMR was exemplified by the overexpression of the NK cell cytotoxicity pathway gene set and, surprisingly, molecularly more like TCMR than active-ABMR., (Copyright © 2023 International Society of Nephrology. Published by Elsevier Inc. All rights reserved.)

RNA-sequencing (RNA-seq) is a technique to determine the order of nucleotides in an RNA segment. Modern sequencing platforms simultaneously sequence millions of RNA molecules. Advances in bioinformatics have allowed us to collect, store, analyze, and disseminate data from RNA-seq experiments and decipher biological insights from large sequencing datasets. Although bulk RNA-seq has significantly advanced our understanding of tissue-specific gene expression and regulation, recent advances in single-cell RNA-seq have allowed such information to be mapped to individual cells, thus remarkably enhancing our insight into discrete cellular functions within a biospecimen. These different RNA-seq experimental approaches require specialized computational tools. Herein, we will first review the RNA-seq experimental workflow, discuss the common terminologies used in RNA-seq, and suggest approaches for standardization across multiple studies. Next, we will provide an up-to-date appraisal of the applications of bulk RNA-seq and single-cell/nucleus RNA-seq in preclinical and clinical research on kidney transplantation, as well as typical bioinformatic workflows utilized in such analysis. Lastly, we will deliberate on the limitations of this technology in transplantation research and briefly summarize newer technologies that could be combined with RNA-seq to permit more powerful dissections of biological functions. Because each step in RNA-seq workflow has numerous variations and could potentially impact the results, as conscientious citizens of the research community, we must strive to continuously modernize our analytical pipelines and exhaustively report their technical details., Competing Interests: The authors declare no conflicts of interest., (Copyright © 2023 Wolters Kluwer Health, Inc. All rights reserved.)

Background: In 2021, four patients who had received solid organ transplants in the USA developed encephalitis beginning 2-6 weeks after transplantation from a common organ donor. We describe an investigation into the cause of encephalitis in these patients., Methods: From Nov 7, 2021, to Feb 24, 2022, we conducted a public health investigation involving 15 agencies and medical centres in the USA. We tested various specimens (blood, cerebrospinal fluid, intraocular fluid, serum, and tissues) from the organ donor and recipients by serology, RT-PCR, immunohistochemistry, metagenomic next-generation sequencing, and host gene expression, and conducted a traceback of blood transfusions received by the organ donor., Findings: We identified one read from yellow fever virus in cerebrospinal fluid from the recipient of a kidney using metagenomic next-generation sequencing. Recent infection with yellow fever virus was confirmed in all four organ recipients by identification of yellow fever virus RNA consistent with the 17D vaccine strain in brain tissue from one recipient and seroconversion after transplantation in three recipients. Two patients recovered and two patients had no neurological recovery and died. 3 days before organ procurement, the organ donor received a blood transfusion from a donor who had received a yellow fever vaccine 6 days before blood donation., Interpretation: This investigation substantiates the use of metagenomic next-generation sequencing for the broad-based detection of rare or unexpected pathogens. Health-care workers providing vaccinations should inform patients of the need to defer blood donation for at least 2 weeks after receiving a yellow fever vaccine. Despite mitigation strategies and safety interventions, a low risk of transfusion-transmitted infections remains., Funding: US Centers for Disease Control and Prevention (CDC), the Biomedical Advanced Research and Development Authority, and the CDC Epidemiology and Laboratory Capacity Cooperative Agreement for Infectious Diseases., Competing Interests: Declaration of interests LFW received research funding from Accelerate Diagnostics, bioMérieux, Hardy Diagnostics, Roche Molecular Systems, and Selux Diagnostics and honoraria from Roche Molecular Systems, Shionogi, and Talis Biomedical, all unrelated to this work. KSG received research support from ThermoFisher and has a royalty-generating collaborative agreement with ZeptoMetrix, both unrelated to this work. MRW received research grant funding from Roche/Genentech and Novartis and speaking honoraria from Genentech, Novartis, Takeda, and WebMD, all unrelated to this work. CYC received research grant funding from the Bay Area Lyme Disease Foundation and the Chan-Zuckerberg Biohub, unrelated to this work, and is on the scientific advisory board for Mammoth Biosciences, Poppy Health, and BiomeSense. MRW and CYC are consultants and co-founders of Delve Bio. CYC is a co-inventor on US patent 11380421, Pathogen Detection Using Next Generation Sequencing, under which algorithms for taxonomic classification, filtering, and pathogen detection are used by SURPI+ software., (Copyright © 2023 The Author(s). Published by Elsevier Ltd. This is an Open Access article under the CC BY-NC-ND 4.0 license. Published by Elsevier Ltd.. All rights reserved.)

Chronic allograft dysfunction (CAD), characterized histologically by interstitial fibrosis and tubular atrophy, is the major cause of kidney allograft loss. Here, using single nuclei RNA sequencing and transcriptome analysis, we identified the origin, functional heterogeneity, and regulation of fibrosis-forming cells in kidney allografts with CAD. A robust technique was used to isolate individual nuclei from kidney allograft biopsies and successfully profiled 23,980 nuclei from five kidney transplant recipients with CAD and 17,913 nuclei from three patients with normal allograft function. Our analysis revealed two distinct states of fibrosis in CAD; low and high extracellular matrix (ECM) with distinct kidney cell subclusters, immune cell types, and transcriptional profiles. Imaging mass cytometry analysis confirmed increased ECM deposition at the protein level. Proximal tubular cells transitioned to an injured mixed tubular (MT1) phenotype comprised of activated fibroblasts and myofibroblast markers, generated provisional ECM which recruited inflammatory cells, and served as the main driver of fibrosis. MT1 cells in the high ECM state achieved replicative repair evidenced by dedifferentiation and nephrogenic transcriptional signatures. MT1 in the low ECM state showed decreased apoptosis, decreased cycling tubular cells, and severe metabolic dysfunction, limiting the potential for repair. Activated B, T and plasma cells were increased in the high ECM state, while macrophage subtypes were increased in the low ECM state. Intercellular communication between kidney parenchymal cells and donor-derived macrophages, detected several years post-transplantation, played a key role in injury propagation. Thus, our study identified novel molecular targets for interventions aimed to ameliorate or prevent allograft fibrogenesis in kidney transplant recipients., (Copyright © 2023 International Society of Nephrology. Published by Elsevier Inc. All rights reserved.)

Drought is a significant abiotic stress that alters plant physiology and ultimately affects crop productivity. Among essential plant nutrients, potassium (K) is known to mitigate the deleterious effect of drought on plant growth. If so, K addition or inoculation of potassium solubilizing microorganisms (KSMs) that are tolerant to drought should promote plant growth during water stress. Therefore, in this study, K solubilizing Aspergillus violaceofuscus and Bacillus licheniformis , isolated from saxicolous environments, were tested for their capacity to tolerate drought using different molecular weights (~4000, 6000, and 8000 Da), and concentrations (0, 250, 500, 750, 1000, and 1250 mg/L) of polyethylene glycol (PEG) under in vitro conditions. The results showed that high concentrations (750 and 1000 mg/L) of PEG with different molecular weight considerably improved bacterial cell numbers/fungal biomass and catalase (CAT) and proline activities. Moreover, the ability of KSMs alone or in combination to impart drought tolerance and promote plant growth in the presence and absence of mica (9.3% K 2 O) supplementation was tested in Alfisol and Vertisol soil types under greenhouse conditions. The results revealed that the tomato plants inoculated with KSMs individually or dually with/without mica improved the physiological and morphological traits of the tomato plants under drought. Generally, tomato plants co-inoculated with KSMs and supplemented with mica were taller (2.62 and 3.38-fold) and had more leaf area (2.03 and 1.98-fold), total root length (3.26 and 8.86-fold), shoot biomass (3.87 and 3.93-fold), root biomass (9.00 and 7.24-fold), shoot K content (3.08 and 3.62-fold), root K content (3.39 and 2.03-fold), relative water content (1.51 and 1.27-fold), CAT activity (2.11 and 2.14-fold), proline content (3.41 and 3.28-fold), and total chlorophyll content (1.81 and 1.90-fold), in unsterilized Alfisol and Vertisol soil types, respectively, than uninoculated ones. Dual inoculation of the KSMs along with mica amendment, also improved the endorrhizal symbiosis of tomato plants more than their individual inoculation or application in both soil types. These findings imply that the A. violaceofuscus and B. licheniformis isolates are promising as novel bioinoculants for improving crop growth in water-stressed and rainfed areas of the tropics in the future., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Muthuraja, Muthukumar and Natthapol.)

Rationale & Objective: Donor acute kidney injury (AKI) activates innate immunity, enhances HLA expression in the kidney allograft, and provokes recipient alloimmune responses. We hypothesized that injury and inflammation that manifested in deceased-donor urine biomarkers would be associated with higher rates of biopsy-proven acute rejection (BPAR) and allograft failure after transplantation., Study Design: Prospective cohort., Setting & Participants: 862 deceased donors for 1,137 kidney recipients at 13 centers., Exposures: We measured concentrations of interleukin 18 (IL-18), kidney injury molecule 1 (KIM-1), and neutrophil gelatinase-associated lipocalin (NGAL) in deceased donor urine. We also used the Acute Kidney Injury Network (AKIN) criteria to assess donor clinical AKI., Outcomes: The primary outcome was a composite of BPAR and graft failure (not from death). A secondary outcome was the composite of BPAR, graft failure, and/or de novo donor-specific antibody (DSA). Outcomes were ascertained in the first posttransplant year., Analytical Approach: Multivariable Fine-Gray models with death as a competing risk., Results: Mean recipient age was 54 ± 13 (SD) years, and 82% received antithymocyte globulin. We found no significant associations between donor urinary IL-18, KIM-1, and NGAL and the primary outcome (subdistribution hazard ratio [HR] for highest vs lowest tertile of 0.76 [95% CI, 0.45-1.28], 1.20 [95% CI, 0.69-2.07], and 1.14 [95% CI, 0.71-1.84], respectively). In secondary analyses, we detected no significant associations between clinically defined AKI and the primary outcome or between donor biomarkers and the composite outcome of BPAR, graft failure, and/or de novo DSA., Limitations: BPAR was ascertained through for-cause biopsies, not surveillance biopsies., Conclusions: In a large cohort of kidney recipients who almost all received induction with thymoglobulin, donor injury biomarkers were associated with neither graft failure and rejection nor a secondary outcome that included de novo DSA. These findings provide some reassurance that centers can successfully manage immunological complications using deceased-donor kidneys with AKI., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Background: We developed urinary cell mRNA profiling for noninvasive diagnosis of acute T cell mediated rejection (TCMR) and BK virus nephropathy (BKVN), two significant post-transplant complications. Our profiling protocol for the multicenter Clinical Trial of Transplantation-04 (CTOT-04) study consisted of centrifugation of urine to prepare cell pellets, washes, addition of an RNA preservative, storage at 80 0 C and shipment in cold containers to our Gene Expression Monitoring (GEM) Core for RNA isolation and quantification of mRNA in RT-qPCR assays. To simplify profiling, we developed a filter-based protocol (ZFBP) that eliminated the need for centrifugation, RNA preservative, storage at 80 0 C, and shipment in cold containers for mRNA profiling. Furthermore, we trained kidney allograft recipients to perform the filtration of urine at home using the filter and post the urinary cell lysate containing the RNA at ambient temperature to our GEM Core for profiling. Here, we report our refinement of ZFBP and investigation of its diagnostic performance characteristics., Methods: Total RNA was isolated from kidney allograft biopsy-matched urines using a filter-based protocol complemented by a silica-membrane-based cartridge for mRNA enrichment, the Weill Cornell Hybrid Protocol (WCHP). Absolute copy numbers of CD3ε mRNA, CXCL10 mRNA, and 18S rRNA, components of the CTOT-04 three-gene TCMR diagnostic signature, and urinary cell BKV VP 1 mRNA copy number were measured using RT-qPCR assays. Mann-Whitney test, Fischer exact test, and receiver operating characteristic (ROC) curve analysis were used for data analyses., Results: Urinary cell three-gene TCMR diagnostic signature scores in urines processed using the WCHP discriminated kidney allograft recipients with TCMR (12 TCMR biopsies from 11 patients) from those without TCMR or BKVN (29 No TCMR/No BKVN biopsies from 29 patients). The median (25th and 75th percentiles) score of the CTOT-04 three-gene TCMR diagnostic signature was -0.448 (-1.664, 0.204) in the TCMR group and - 2.542 (-3.267, -1.365) in the No TCMR/ No BKVN group (P = 0.0005, Mann-Whitney test). ROC curve analysis discriminated the TCMR group from the No TCMR/ No BKVN group; the area under the ROC curve (AUROC) was 0.84 (95% Confidence Intervals [CI], 0.69 to 0.98) (P < 0.001), and TCMR was diagnosed with a sensitivity of 67% (95% CI, 35 to 89) at a specificity of 86% (95% CI, 67 to 95) using the CTOT-04 validated cutpoint of -1.213 (P = 0.0016, Fisher exact test). BKV VP1 mRNA copy number in urines processed using the WCHP discriminated patients with BKVN (n = 7) from patients without TCMR or BKVN (n = 29) and the AUROC was 1.0 (95% CI, 1.00 to 1.00) (P < 0.0001) and BKVN was diagnosed with a sensitivity of 86% (95% CI, 42 to 99) at a specificity of 100% (95% CI, 85 to 100) with the previously validated cutpoint of 6.5 × 10 8 BKV-VP1 mRNA copies per microgram of RNA (P < 0.0001, Fisher exact test)., Conclusion: Urine processed using the WCHP predicted TCMR and BKVN in kidney allograft recipients. WCHP represents not only a significant advance toward the portability of urinary cell mRNA profiling but also improved patient management by minimizing their visits for urine collection., (Copyright © 2022. Published by Elsevier B.V.)

Background: Kidney transplantation in HIV-infected individuals with end-stage kidney disease is associated with improved survival compared to dialysis. Rabbit anti-thymocyte globulin (rATG) induction in HIV-infected kidney transplant recipients has been associated with a lower risk of acute rejection, but data on the rates of de novo malignancy and BK viremia in these patients is lacking., Methods: We performed a single-center retrospective cohort study of adult HIV-infected individuals who underwent kidney transplantation with rATG induction between January 2006 and December 2016. The primary outcome was the development of de novo malignancy. Secondary outcomes included the development of BK viremia, infections requiring hospitalization, HIV progression, biopsy-proven acute rejection, and patient and allograft survival., Results: Twenty-seven HIV-infected individuals with end-stage kidney disease received deceased (n=23) or living (n=4) donor kidney transplants. The cumulative rate of malignancy at five years was 29%, of whom 29% died because of advanced malignancy. BK viremia was detected in six participants (22%), of whom one had biopsy-proven BK virus-associated nephropathy and all of whom cleared the BK viremia. Five-year acute rejection rates, patient survival and death-censored allograft survival were 17%, 85% and 80% respectively., Conclusion: rATG induction in HIV-infected kidney transplant recipients was associated with a low risk of acute rejection, but a potentially higher risk of de novo malignancies and BK viremia in this cohort. Screening strategies to closely monitor for BK virus infection and malignancy post-transplantation may improve outcomes in HIV-infected kidney transplant recipients receiving rATG induction., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Al Jurdi, Liu, Salinas, Aull, Lubetzky, Drelick, Small, Kapur, Hartono and Muthukumar.)

Gloriosa superba L. is of great economic importance due to its high medicinal value. Nevertheless, there is a need to reexamine species delimitation in the Gloriosa taxa as most of the species have been synonymised as G. superba. Therefore, the present study was undertaken to investigate the vegetative anatomical traits of G. superba. The leaf, scale leaf, tendril, stem, tuber, and roots of G. superba were freehand sectioned and stained with various staining solutions to record the anatomical structures. The cellular dimensions of each plant part were measured. The present study revealed the presence of intercostal and costal regions in the leaf epidermis, anomocytic stomata on abaxial surface, uniseriate epidermis covered by cuticle, undifferentiated mesophyll, and a bundle sheath surrounding vascular bundles in a leaf. Unlike the leaf, the scale leaf contains air chambers in the mesophyll region and bundle sheath is absent. The tendril had uniseriate cuticularized epidermis followed by few layers of cells developing wall thickenings, and collateral vascular bundles. The mature stem is differentiated from the young stem by the presence of bi-layered epidermis, the absence of stomata on the stem surface, and chlorenchymatous hypodermis. Air passage containing epidermis covered by thin cuticle is recorded in the stem. Starch grains are present in the tuber ground tissue. Velamen is reported for the first time in G. superba root. Scalariform perforation end plate present in root metaxylem. Roots of G. superba are colonized by arbuscular mycorrhizal and dark septate endophytic fungi. Therefore, these anatomical traits could aid in the identification of G. superba. RESEARCH HIGHLIGHTS: Anatomy of vegetative parts of Gloriosa superba was studied. Air-passage enveloped by uniseriate epidermis present in stem. Bundle sheath surrounds vascular bundles of leaf and stem. Cells of rhizome ground tissue contain abundant starch grains. Velamen tissue is reported for the first time in roots., (© 2022 Wiley Periodicals LLC.)

The vegetative anatomy of Elaeocarpus angustifolius Blume, Elaeocarpus tuberculatus Roxb., and Elaeocarpus variabilis Zmarzty were investigated to illustrate anatomical variations. Plant materials were free-hand sectioned using a razor blade and stained with different staining solutions. The maceration technique was used to assess stomatal characteristics. Elaeocarpus leaves have abaxial epidermis with paracytic stomata and curved anticlinal walls in E. angustifolius, straight walls in other two species. Trichomes were absent in E. angustifolius.hav Mesophyll dorsiventral, midvein cortex contains starch grains, and vascular tissues enclosed by thick-walled sclerenchymatous cells. The petioles of all the three species possess unicellular epidermal hairs, collenchymatous hypodermis, and cortex containing druses and crystals, and vascular tissue enclosed by sclerenchymatous fibers. Water-storage cells are absent in petioles of E. angustifolius. Anatomical features of Elaeocarpus stem include epidermal hairs, epidermis covered by thin cuticle, the collenchymatous hypodermis and vascular integrity with entire cylinder enclosed by sclerenchymatous fibers. Pith contains water-storage cells. Starch grains absent in the pith cells of E. tuberculatus. The roots of Elaeocarpus possess unicellular root hairs, cortex 12-14 layered in E. tuberculatus and E. variabilis and 10-12 layerd E. angustifolius, Endodermis O-thickened and pericycle single-layered in all the examined Elaeocarpus species. Vascular bundles are arranged radially. Lignin deposition occurred in stellar region of roots. Water-storage cells present in the stelar regions of E. variabilis. The study revealed significant anatomical differences between the three Elaeocarpus species and most of these anatomical features may be used as markers for the identification of these species. RESEARCH HIGHLIGHTS: Comparative anatomy of three south Indian Elaeocarpus was studied. Leaf mesophyll layers varied in all the Elaeocarpus species. Crystals was present in petiole of all examined Elaeocarpus species. Starch grains was absent in stems of E. tubercuatus, but present in E. variabilis. Water-storage cells observed in stellar region of E. variabilis., (© 2022 Wiley Periodicals LLC.)

We tested the hypothesis that single-cell RNA-sequencing (scRNA-seq) analysis of human kidney allograft biopsies will reveal distinct cell types and states and yield insights to decipher the complex heterogeneity of alloimmune injury. We selected 3 biopsies of kidney cortex from 3 individuals for scRNA-seq and processed them fresh using an identical protocol on the 10x Chromium platform; (i) HK: native kidney biopsy from a living donor, (ii) AK1: allograft kidney with transplant glomerulopathy, tubulointerstitial fibrosis, and worsening graft function, and (iii) AK2: allograft kidney after successful treatment of active antibody-mediated rejection. We did not study T-cell-mediated rejections. We generated 7217 high-quality single cell transcriptomes. Taking advantage of the recipient-donor sex mismatches revealed by X and Y chromosome autosomal gene expression, we determined that in AK1 with fibrosis, 42 months after transplantation, more than half of the kidney allograft fibroblasts were recipient-derived and therefore likely migratory and graft infiltrative, whereas in AK2 without fibrosis, 84 months after transplantation, most fibroblasts were donor-organ-derived. Furthermore, AK1 was enriched for tubular progenitor cells overexpressing profibrotic extracellular matrix genes. AK2, eight months after successful treatment of rejection, contained plasmablast cells with high expression of immunoglobulins, endothelial cell elaboration of T cell chemoattractant cytokines, and persistent presence of cytotoxic T cells. In addition to these key findings, our analysis revealed unique cell types and states in the kidney. Altogether, single-cell transcriptomics yielded novel mechanistic insights, which could pave the way for individualizing the care of transplant recipients., Competing Interests: The authors have declared that no competing interests exist.

Soil salinity is an important stress that negatively affects crop growth and productivity, causing extensive agricultural losses, worldwide. Potassium (K) solubilizing microorganisms (KSMs) can impart abiotic stress tolerance in plants in addition to nutrient solubilization. In this study, the salinity tolerance of KSMs Bacillus licheniformis and Aspergillus violaceofuscus originating from saxicolous habitats was examined using different concentrations of NaCl (0, 25, 50, 75, 100, and 125 mM) under in vitro conditions. The results indicated that both KSMs were capable of tolerating salinity. As B. licheniformis had a maximum growth in 100 mM NaCl at 37 °C, A. violaceofuscus had the maximum biomass and catalase (CAT) activity at 75 mM NaCl. However, maximum proline content was detected at 100 mM NaCl in both KSMs. Further, the ability of these KSMs to promote tomato growth individually and in combination with the presence or absence of mica was also examined in unsterilized or sterilized Alfisol and Vertisol soils under induced salinity in greenhouse conditions. The results of the greenhouse study revealed that inoculation of KSMs along with/without mica amendment significantly improved the morphological and physiological characteristics of tomato plants under salinity. Plant height, leaf area, biomass, relative water content, proline content, and CAT activity of dual inoculated plants were significantly higher than non-inoculated plants. Significant correlations existed between various soil, plant growth, soil pH and available K. From the results, it could be concluded that B. licheniformis and A. violaceofuscus are potential candidates for improving crop production in saline-stressed soils., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Macrophages exert critical functions during kidney injury, inflammation, and tissue repair or fibrosis. Mitochondrial structural and functional aberrations due to an imbalance in mitochondrial fusion/fission processes are implicated in the pathogenesis of chronic kidney disease. Therefore, we investigated macrophage-specific functions of mitochondrial fusion proteins, mitofusin (MFN)1 and MFN2, in modulating macrophage mitochondrial dynamics, biogenesis, oxidative stress, polarization, and fibrotic response. MFN1 and MFN2 were found to be suppressed in mice after adenine diet-induced chronic kidney disease, in transforming growth factor-beta 1-treated bone marrow-derived macrophages, and in THP-1-derived human macrophages (a human leukemic cell line). However, abrogating Mfn2 but not Mfn1 in myeloid-lineage cells resulted in greater macrophage recruitment into the kidney during fibrosis and the macrophage-derived fibrotic response associated with collagen deposition culminating in worsening kidney function. Myeloid-specific Mfn1 /Mfn2 double knockout mice also showed increased adenine-induced fibrosis. Mfn2-deficient bone marrow-derived macrophages displayed enhanced polarization towards the profibrotic/M2 phenotype and impaired mitochondrial biogenesis. Macrophages in the kidney of Mfn2-deficient and double knockout but not Mfn1-deficient mice exhibited greater mitochondrial mass, size, oxidative stress and lower mitophagy under fibrotic conditions than the macrophages in the kidney of wild-type mice. Thus, downregulation of MFN2 but not MFN1 lead to macrophage polarization towards a profibrotic phenotype to promote kidney fibrosis through a mechanism involving suppression of macrophage mitophagy and dysfunctional mitochondrial dynamics., (Copyright © 2022 International Society of Nephrology. Published by Elsevier Inc. All rights reserved.)

The human population is increasing by 0.96% annually and is estimated to reach from 7.3 to 9 billion in 2050 and 11 billion in 2100. The world's agriculture is under pressure to produce more food and ensure food security. On the other hand, around 40% of the cultivable land is already degraded due to various factors including urbanization, soil sealing, soil acidification, salinization, soil erosion, and contamination. Arbuscular mycorrhizal fungi (AMF) constitute a unique group of root obligate symbiont that exchange mutual benefits with about 90% of terrestrial plants and represents a key link between plants and soil mineral nutrients. Literature is scanty on the studies on massive inoculation of AMF in food crops in agronomic settings, and thereby achieving efficient uptake and minimization of the major soil nutrients, eventually meeting our food demand under increasing and inevitable stressed environments. Given above, this review aimed to (i) introduce agricultural soil-contamination, and the relation of soil microbiome with the health of soils and plants; (ii) briefly overview AMF; (iii) highlight AMF role as a bioinoculant, and enhancer of efficient uptake and loss-minimization of nutrients; (iv) appraise literature available on AMF role in the regulation of growth and nutrition mainly in vegetable, horticultural crops and fruit trees; (v) enlighten the role and major mechanisms underlying AMF-mediated regulation of plant growth and nutrition under major biotic and abiotic stresses; (vi) highlight AMF role in the minimization of greenhouse gas emissions; and (vii) list major aspects so far unexplored in the current context., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Background: Acute rejection (AR) and recurrent hepatitis C virus (R-HCV) are significant complications in liver allograft recipients. Noninvasive diagnosis of intragraft pathologies may improve their management., Methods: We performed small RNA sequencing and microRNA (miRNA) microarray profiling of RNA from sera matched to liver allograft biopsies from patients with nonimmune, nonviral (NINV) native liver disease. Absolute levels of informative miRNAs in 91 sera matched to 91 liver allograft biopsies were quantified using customized real-time quantitative PCR (RT-qPCR) assays: 30 biopsy-matched sera from 26 unique NINV patients and 61 biopsy-matched sera from 41 unique R-HCV patients. The association between biopsy diagnosis and miRNA abundance was analyzed by logistic regression and calculating the area under the receiver operating characteristic curve., Results: Nine miRNAs-miR-22, miR-34a, miR-122, miR-148a, miR-192, miR-193b, miR-194, miR-210, and miR-885-5p-were identified by both sRNA-seq and TLDA to be associated with NINV-AR. Logistic regression analysis of absolute levels of miRNAs and goodness-of-fit of predictors identified a linear combination of miR-34a + miR-210 (P < 0.0001) as the best statistical model and miR-122 + miR-210 (P < 0.0001) as the best model that included miR-122. A different linear combination of miR-34a + miR-210 (P < 0.0001) was the best model for discriminating NINV-AR from R-HCV with intragraft inflammation, and miR-34a + miR-122 (P < 0.0001) was the best model for discriminating NINV-AR from R-HCV with intragraft fibrosis., Conclusions: Circulating levels of miRNAs, quantified using customized RT-qPCR assays, may offer a rapid and noninvasive means of diagnosing AR in human liver allografts and for discriminating AR from intragraft inflammation or fibrosis due to R-HCV., Competing Interests: T.T. is a cofounder of and scientific advisor to Alnylam Pharmaceuticals and a scientific advisor to Regulus Therapeutics. M.S. has a Consultancy Agreement with CareDx, Inc. Brisbane, CA and with Sparks Therapeutics, Philadelphia, PA. The other authors of this article declare no conflicts of interest. The other authors declare no conflict of interests., (Copyright © 2021 Wolters Kluwer Health, Inc. All rights reserved.)

Human leukocyte antigens (HLAs) are the primary determinants of alloimmunity. A crossmatch test is a test that determines the immunologic risk of a recipient with a potential donor by ensuring that there are no transplant-relevant circulating antibodies in the recipient directed against donor antigens. Physical crossmatch (PXM) tests, such as complement-dependent cytotoxicity crossmatch (CDCXM) and flow cytometry crossmatch (FCXM), require mixing of patient serum and donor cells, are labor intensive, and are logistically challenging. Virtual crossmatch (VXM) test assesses immunologic compatibility between recipient and potential donor by analyzing the results of 2 independently done physical laboratory tests-patient anti-HLA antibody and donor HLA typing. The goal of VXM is pretransplant risk stratification-though there is no consensus on whether such risk assessment involves predicting the PXM result or the posttransplant outcome. Although the concept of VXM is not new, the advent of solid-phase assays for detecting circulating antibodies in the recipient directed against individual HLA and DNA-based methods for typing donor HLA specificities at a higher resolution makes the routine use of VXM a reality. Accordingly, VXM may be applied at different scenarios-both for sensitized and nonsensitized patients. Implementation of VXM-based approach has resulted in statistically significant reduction in cold ischemia time without an increase in hyperacute rejection episodes. Though there are considerable challenges, VXM is expected to be used more often in the future, depending on the transplant center's tolerance of immunologic risk., (© 2022 International Society of Nephrology. Published by Elsevier Inc.)

Background: Acute kidney injury (AKI) in deceased donors is not associated with graft failure (GF). We hypothesize that hemodynamic AKI (hAKI) comprises the majority of donor AKI and may explain this lack of association., Methods: In this ancillary analysis of the Deceased Donor Study, 428 donors with available charts were selected to identify those with and without AKI. AKI cases were classified as hAKI, intrinsic (iAKI), or mixed (mAKI) based on majority adjudication by three nephrologists. We evaluated the associations between AKI phenotypes and delayed graft function (DGF), 1-year eGFR and GF. We also evaluated differences in urine biomarkers among AKI phenotypes., Results: Of the 291 (68%) donors with AKI, 106 (36%) were adjudicated as hAKI, 84 (29%) as iAKI and 101 (35%) as mAKI. Of the 856 potential kidneys, 669 were transplanted with 32% developing DGF and 5% experiencing GF. Median 1-year eGFR was 53 (IQR: 41-70) ml/min/1.73m2. Compared to non-AKI, donors with iAKI had higher odds DGF [aOR (95%CI); 4.83 (2.29, 10.22)] and had lower 1-year eGFR [adjusted B coefficient (95% CI): -11 (-19, -3) mL/min/1.73 m2]. hAKI and mAKI were not associated with DGF or 1-year eGFR. Rates of GF were not different among AKI phenotypes and non-AKI. Urine biomarkers such as NGAL, LFABP, MCP-1, YKL-40, cystatin-C and albumin were higher in iAKI., Conclusion: iAKI was associated with higher DGF and lower 1-year eGFR but not with GF. Clinically phenotyped donor AKI is biologically different based on biomarkers and may help inform decisions regarding organ utilization., Competing Interests: The authors have declared that no competing interests exist.