Renal Research Articles

TRPML-1 Dysfunction and Renal Tubulopathy in Mucolipidosis Type IV.

Loss of function mutations in the lysosomal channel TRPML-1 cause mucolipidosis type IV (MLIV), a rare lysosomal storage disease characterized by neurological defects, progressive vision loss, and achlorhydria. Recent reports have highlighted kidney disease and kidney failure in patients with MLIV during the second to third decade of life; however, the molecular mechanisms driving kidney dysfunction remain poorly understood. A cross-sectional review of medical records from 21 MLIV patients (ages 3-43 years) was conducted to assess kidney function impairment. Additionally, we examined the kidney phenotype of MLIV mice at various ages, along with human kidney cells silenced for TRPML-1 and primary tubular cells from wild-type and MLIV mice. Immunohistology and cell biology approaches were used to phenotype nephron structure, the endolysosomal compartment, and inflammation. Kidney function was assessed through proteomic analysis of mouse urine and in vivo renal filtration measurements. Of the 21 MLIV patients only adults were diagnosed with stage 2-3 chronic kidney disease. Laboratory abnormalities included decreased eGFR, higher levels BUN/Creatine in bloodand proteinuria. In MLIV mice, we observed significant alterations in endolysosomal morphology, function, and impaired autophagy in proximal and distal tubules. This led to the accumulation of megalin (LRP2) in the subapical region of proximal tubular cells, indicating a block in apical receptor-mediated endocytosis. In vivo and in vitro experiments confirmed reduced fluid-phase endocytosis and impaired uptake of ligands, including β-lactoglobulin, transferrin, and albumin in MLIV proximal tubular cells. Urine analysis revealed tubular proteinuria and enzymuria in mice with MLIV. Additionally, early-stage disease was marked by increased inflammatory markers, fibrosis, and activation of the pro-inflammatory transcription factor NF-κB, coinciding with endolysosomal defects. Importantly, AAV-mediated TRPML-1 gene delivery reversed key pathological phenotypes in Mucolipidosis type IV mice, underscoring TRPML-1's critical role in kidney function. Our findings link TRPML-1 dysfunction to the development of kidney disease in MLIV, providing new insights into its pathogenesis and potential therapeutic targets.

Apoc1 Knockdown Alleviates High Glucose-induced Oxidative Stress and Apoptosis of Renal Tubular Cells by Binding to Clusterin.

Diabetic nephropathy (DN) is a serious diabetic complication. Renal tubular damage is an important aspect of DN. Increased apolipoprotein C1 (Apoc1) has been confirmed in serum of patients with DN. The exact mechanism of Apoc1 in DN is unclear as yet. We aimed to elaborate the molecular mechanism underlying high glucose (HG)-induced renal tubular epithelial damage. In this content, a DN mouse model was established to assess renal damage. Apoc1 and Clusterin expression in renal tissue was detected using immunoblotting and immunofluorescence staining. In vitro, human kidney proximal tubular epithelial cells (HK-2 cells) were exposed to HG to simulate the DN model. After Apoc1 and/or Clusterin knockdown, HK-2 cell viability under HG conditions was detected using CCK-8 assay. DCFH-DA staining was used to examine the production of intracellular reactive oxygen species (ROS). MDA and SOD levels were tested by kits. Moreover, cell apoptosis was measured using TUNEL staining. Immunoblotting was employed to evaluate the expression of proteins. Additionally, the binding between Apoc1 and Clusterin was analyzed using co-immunoprecipitation experiments. Our data revealed that Apoc1 expression was upregulated while Clusterin expression was downregulated in renal tissue of DN mice and HG-treated HK-2 cells. Apoc1 knockdown alleviated oxidative stress and apoptosis in HG-treated HK-2 cells. Importantly, Apoc1 could bind to Clusterin and regulate Clusterin expression in HK-2 cells. Finally, Clusterin silencing blocked the influences of Apoc1 knockdown on the oxidative stress and apoptosis in HK-2 cells under HG conditions. Collectively, Apoc1 knockdown exerts potential anti-DN effects by binding to Clusterin to alleviate HG-induced renal tubular damage, suggesting that Apoc1/Clusterin can be used as a valuable therapeutic target for DN.

Single-cell transcriptome analysis reveals status changes of immune cells in chronic kidney disease

Background and aimsThe immune system plays a crucial role in the development of kidney diseases. Chronic kidney disease (CKD) can lead to various complications, potentially affecting multiple systems throughout the body. Currently, the description of the immune system in human CKD is not comprehensive enough. Constructing a CKD kidney atlas using single-cell RNA sequencing (scRNA-seq) can provide deeper insights into the composition and functional changes of immune cells in CKD, facilitating the discovery of new therapeutic targets.MethodsWe processed and integrated scRNA-seq datasets from healthy and CKD kidneys from three independent cohorts using the same approach (including 42 normal samples and 23 chronic kidney disease samples). Subsequently, we conducted gene enrichment and intercellular communication analysis to construct an immune cell atlas of the kidneys in CKD patients.ResultsWe identified nine major kidney cell clusters. Further clustering analysis of different immune cell clusters revealed that, compared to normal kidneys, CKD patients’ kidneys had decreased CD16+ NK cells while CD4+ naive helper T cells and CCR7+ DC increased. Partial activation of the WNT signaling pathway was observed in T cells and NK cells of CKD patients, while some metabolism-related genes were inhibited. Myeloid cell subgroups also exhibited abnormal signaling pathway alterations. Additionally, we discovered a unique population of SPP1 macrophages in CKD, which are recruited by chemokines released from aPT and aTAL cell subpopulations. These SPP1 macrophages may promote cellular fibrosis through the signaling of SPP1, FN1, and various receptors.ConclusionWe established a human CKD kidney immune cell atlas and identified SPP1 macrophages as a unique cell type in CKD. The interaction between SPP1 macrophages and damaged cells may serve as a potential therapeutic target for treating CKD in the future.

Polyethylene glycol-phospholipid functionalized single-walled carbon nanotubes for enhanced siRNA systemic delivery

Small interfering RNAs (siRNA) technology has emerged as a promising therapeutic tool for human health conditions like cancer due to its ability to regulate gene silencing. Despite FDA-approved, their delivery remains localized and limiting their systemic use. This study used single-walled carbon nanotubes (SWNTs) functionalized with polyethylene glycolated (PEGylated) phospholipids (PL-PEG) derivatives for systemic siRNA delivery. We developed an siRNA systemic delivery vehicle (SWNT-siRNA) by conjugating SWNT functionalized with PL-PEG containing either amine (PA) or maleimide (MA). The functionalized SWNT with a lower molecular weight of PA produced the SWNT-siRNA conjugate system with the highest stability and high siRNA loading quantity. The system delivered siRNA to a panel of tumour cell lines of different organs (i.e. HeLa, H1299 and MCF-7) and a non-cancerous human embryonic kidney 293 cells (HEK293T) with high biocompatibility and low toxicity. The cellular uptake of SWNT-siRNA conjugates by epithelial cells was found to be energy dependent. Importantly, the presence of P-glycoprotein, a marker for drug resistance, did not inhibit SWNT-mediated siRNA delivery. Mouse xenograft model further confirmed the potential of SWNT-siRNA conjugates with a significant gene knock-down without signs of acute toxicity. These findings pave the way for potential gene therapy applications using SWNTs as delivery vehicles.

Probabilistic Health Risk Assessment of Heavy Metals in People Living Near the Mine-Closure in Tak Province

Heavy metals including lead (Pb), zinc (Zn), and cadmium (Cd) can damage the kidney, liver, brain, lungs, blood composition, and other vital organs. As a result, their toxicity, especially when contaminated in food, water, and soil, can be harmful to human health. This study evaluated the risk of adverse health effects to residents near closed zinc mines in Tak Province, Thailand. Data was collected from three subdistrict sites and analyzed for probable health risk. Scenario pathways were oral and dermal contact of heavy metals; including Pb, Zn, and Cd, in four age groups with average daily dose and hazard quantification; hazard index (HI); and cancer risk (CR) considered. Results were that carcinogen risk quantification exceeded the recommended limit (CR ≤10-6) in all scenarios, and the hazard index for non-carcinogenic risk was below the acceptable level (HQ ≤1), in which Cd dominated for all categories. These findings suggest that all medias and food products should be appropriately monitored, especially with carcinogenic surveillance, to mitigate health effects on residents.

Research Progress of Pyroptosis in Renal Diseases.

Kidney diseases, particularly Acute Kidney Injury (AKI) and Chronic Kidney Disease (CKD), are identified as global public health issues affecting millions of individuals. In addition, the frequency of renal diseases in the population has increased dramatically and rapidly in recent years. Renal disorders have become a significant public health burden. The pathophysiology of renal diseases is significantly connected with renal cell death, including apoptosis, necrosis, necroptosis, ferroptosis, pyroptosis, and autophagy, as is now recognized. Unlike other forms of cell death, pyroptosis is a unique planned cell death (PCD). Scientists have proven that pyroptosis is crucial in developing various disorders, and this phenomenon is gaining increasing attention. It is considered a novel method of inflammatory cell death. Intriguingly, inflammation is among the most significant pathological characteristics of renal disease. This study investigates the effects of pyroptosis on Acute Kidney Injury (AKI), Chronic Kidney Disease (CKD), Diabetic Nephropathy (DN), Immunoglobulin A (IgA) Nephropathy, and Lupus Nephritis (LN) to identify novel therapeutic targets for kidney diseases.

Artificial Intelligence-Based Classification of CT Images Using a Hybrid SpinalZFNet.

The kidney is an abdominal organ in the human body that supports filtering excess water and waste from the blood. Kidney diseases generally occur due to changes in certain supplements, medical conditions, obesity, and diet, which causes kidney function and ultimately leads to complications such as chronic kidney disease, kidney failure, and other renal disorders. Combining patient metadata with computed tomography (CT) images is essential to accurately and timely diagnosing such complications. Deep Neural Networks (DNNs) have transformed medical fields by providing high accuracy in complex tasks. However, the high computational cost of these models is a significant challenge, particularly in real-time applications. This paper proposed SpinalZFNet, a hybrid deep learning approach that integrates the architectural strengths of Spinal Network (SpinalNet) with the feature extraction capabilities of Zeiler and Fergus Network (ZFNet) to classify kidney disease accurately using CT images. This unique combination enhanced feature analysis, significantly improving classification accuracy while reducing the computational overhead. At first, the acquired CT images are pre-processed using a median filter, and the pre-processed image is segmented using Efficient Neural Network (ENet). Later, the images are augmented, and different features are extracted from the augmented CT images. The extracted features finally classify the kidney disease into normal, tumor, cyst, and stone using the proposed SpinalZFNet model. The SpinalZFNet outperformed other models, with 99.9% sensitivity, 99.5% specificity, precision 99.6%, 99.8% accuracy, and 99.7% F1-Score in classifying kidney disease.

Coupling a process-driven model with geographic information system enables the spatial predication of cadmium in paddy soils

Policies on the management of paddy fields are usually made at a broad scale and from a long-term perspective, while predicting the spatial extent of cadmium (Cd) contamination in paddy soils remains challenging. In this study, we developed a process-driven spatial model to quantify the transport of Cd in paddy soils and validated it against observed data from a 10-year regional investigation in southern China. Using a geographic information system and Monte Carlo simulation, the model was then applied to evaluate the effectiveness of different remediation strategies for contaminated paddy fields at field-to-regional scales in a 100-year period. In the last decade, atmospheric emissions have accounted for 43.5% of the total Cd input in local paddy soils. However, the local clean air act failed to mitigate Cd contamination in 99.8% of study area over the period of 2020–2120 because straw return became the dominant contributor to Cd inputs. Improving aerosol emission reductions by 3% per year, stopping straw return to soil, and cleaning irrigation channels would take approximately 30 years (2020–2050) to protect 95% of local rice production from causing an excessive human Cd kidney burden, especially in the paddy fields located in mining-affected areas.

MicroRNAs in fluorosis pathogenesis: impact on dental, skeletal, and soft tissues.

Fluoride-induced toxicity (fluorosis) poses a significant health concern globally, affecting millions of individuals. Understanding the molecular mechanisms underlying fluorosis, particularly the role of microRNAs (miRNAs), is crucial for developing effective preventive and therapeutic strategies. This review explores the pivotal role of miRNAs in the pathogenesis of fluorosis, particularly examining its impact on both hard (skeletal and dental) and soft (brain, liver, kidney, heart, and reproductive organs) tissues. Skeletal fluorosis manifests as abnormal bone mineralization and structure, while dental fluorosis affects enamel formation. In vitro and in vivo studies suggest a significant involvement of miRNAs in the progression of these conditions. For skeletal fluorosis, miR-124, miR-155, and miR-200c-3phave been identified askeyregulators, while miR-296-5p and miR-214-3pare implicatedin dental fluorosis.Moreover, soft tissue fluorosis encompasses a spectrum of adverse effects on various organs, including the brain, liver, kidneys, heart, and reproductive system. In soft tissues, miRNAs, such as miR-124, miR-200c-3p, miR-132, and miR-34b-5p, havebeen linkedto cellular damage and dysfunction. Notably, miRNAs exert their effectsthrough the modulation ofcritical pathways involved in fluorosis pathology, including Wnt signaling, apoptosis, cell cycle, and autophagy. Understanding the regulatory roles of miRNAs in fluorosis pathogenesis holds promise for identifying biomarkers and therapeutic targets. However, further research is needed to elucidate the molecular mechanisms underlying miRNA-mediated responses to fluoride exposure.Integration ofmiRNA research into fluorosis studies could facilitate the development of diagnostic tools and therapeutic interventions, thus mitigating the detrimental effects of fluorosis on both hard and soft tissues.

Jia Wei Qingxin Lotus Seed Drink ameliorates epithelial mesenchymal transition injury in diabetic kidney disease via inhibition of JMJD1C/SP1/ZEB1 signaling pathway

BackgroundDiabetic kidney disease (DKD) is one of the most common microvascular complications in patients with diabetes mellitus. In this condition, renal tubular epithelial mesenchymal transition (EMT) is an important factor accelerating the progression of DKD and a major cause of renal fibrosis and end-stage renal disease. However, the therapeutic effect is unsatisfactory because of the lack of effective drugs. Jia Wei Qingxin Lotus Seed Drink (QISD) is a traditional Chinese medicine compound formula that has shown to be effective in the clinical treatment of DKD. However, the potential of QISD in DKD-EMT treatment has yet to be fully explored. PurposeThis study aimed to investigate the role of QISD in ameliorating DKD-EMT injury and its mechanism. MethodsThe active ingredients of QISD were identified via ultra-performance liquid chromatography-mass spectrometry/mass spectrometry (UHPLC-MS/MS). A DKD mouse model was constructed by high-fat diet feeding and intraperitoneal injection of STZ (60 mg/kg), and QISD (14.46, 28.92, and 57.84 g/kg/day) was administered by gavage for 12 consecutive weeks. Dapagliflozin (1 mg/kg/d) was used as a positive control. Renal pathological damage was observed by HE, PAS, and Masson staining. The expression levels of EMT-related proteins and pathway proteins were detected via immunohistochemistry, RT-qPCR, and western blot. In in vitro experiments, EMT injury was induced in human kidney tubular epithelial cells (HK-2) by using lipopolysaccharide (LPS). A combination of CCK8 assay, wound healing assay, small-molecule inhibitor intervention, and overexpression lentiviral transfection was used to investigate the effects of QISD on cell migration ability, adhesion ability, fibrotic factor formation, and mesenchymal properties. ResultsAnimal experiments showed that QISD improved blood glucose, body weight, symptoms of excessive drinking and eating, and renal pathological injury in mice, reduced extracellular matrix deposition, delayed renal EMT injury, and inhibited the activation of the histone demethylase JMJD1C. UHPLC-MS/MS and molecular docking indicated that baicalin, wogonoside, oroxylin A-7-O-β-D-glucuronide, and glulisine A found in QISD could bind to JMJD1C. The ameliorating effect of QISD on DKD-EMT injury might be related to JMJD1C. The improvement of DKD-EMT injury by QISD was accompanied by the reduction of SP1 and ZEB1 expression. The SP1 overexpression not only reversed the therapeutic effect of JIB-04, an inhibitor of JMJD1C, on DKD-EMT but also exacerbated the expression of ZEB1 and downstream EMT-related factors. Thus, QISD might affect the expression of the epithelial marker E-cadherin by inhibiting the JMJD1C/SP1/ZEB1 signaling pathway, consequently preventing the transformation of epithelial cells to mesenchymal cells and ameliorating DKD-EMT injury. ConclusionThis study was the first to demonstrate that QISD might ameliorate DKD-EMT injury by inhibiting the JMJD1C/SP1/ZEB1 signaling pathway. These findings provide strong pharmacologic evidence for the clinical use of QISD in the treatment of DKD.

Toxicological risk assessment of triadimenol for human exposure, broiler health, and food safety

Triadimenol, a widely used triazole fungicide, leaves residues that pose risks to broiler health, food safety, and human health. Current studies focus on lab animals, leaving limited data regarding its impact on non-target organisms in agricultural ecosystems. Moreover, the doses in current studies often exceed typical agricultural pollution levels of triadimenol. Therefore, this study evaluates the toxic effects of triadimenol by exposing broilers to different concentrations (0.05-20 mg/kg) in their feed for 42 days, assessing growth performance, organ index, hematological parameters, histopathology, jejunum morphology, and tissue residues. The results show that triadimenol contamination at 0.05-20 mg/kg in feed does not significantly affect broiler growth performance. However, the significant changes in hematological parameters suggest the potential hematological toxicity of triadimenol in broilers. Triadimenol at 1 mg/kg or higher induces hepatotoxic and nephrotoxic effects, and significantly alters kidney organ index and histopathology in broilers. Additionally, when the triadimenol contamination level in feed exceeds 1 mg/kg, the residues in edible tissues of broilers exceed the limits set by the EU and China. Overall, our study indicates that even low-dose exposure to triadimenol poses potential risks, highlighting the need for strict regulation of its use in agriculture to protect food safety and human health.

Abnormalities of IL-12 Family Cytokine Pathways in Autosomal Dominant Polycystic Kidney Disease Progression

Background and Objectives: Autosomal Dominant Polycystic Kidney Disease (ADPKD) is the most frequent genetic renal disease with a complex physiopathology. More and more studies sustain that inflammation plays a crucial role in ADPKD pathogenesis and progression. We evaluated IL-12 involvement in ADPKD pathophysiology by assessing the serum levels of its monomers and heterodimers. Materials and Methods: A prospective case-control study was developed and included 66 ADPKD subjects and a control group of 40 healthy subjects. The diagnosis of ADPKD was based on familial history clinical and imagistic exams. The study included subjects with eGFR > 60 mL/min/1.73 mp, with no history of hematuria or other renal disorders, with stable blood pressure in the last 6 months. We tested serum levels of monomers IL-12 p40 and IL-12 p35 and heterodimers IL-12 p70, IL-23, IL 35, assessed by ELISA method. Results: IL-12 family programming was abnormal in ADPKD patients. IL-12p70, IL-12p40, and IL-23 secretion increased, while IL-12p35 and IL-35 secretion decreased compared to control. IL-12p70, IL-12p40, and IL-23 had a progressive increase correlated with immune response amplification, a decrease of eGFR, an increase in TKV, and in albuminuria. On the other hand, IL-35 and IL-12p35 were correlated negatively with CRP and albuminuria and positively with eGFR in advanced ADPKD. Conclusions: The present study investigated IL-12 cytokine family members’ involvement in ADPKD pathogenesis, enriching our understanding of inflammation in the most common renal genetic disorder.

Analysis of Complement Factor H gene polymorphisms and their association with clinical manifestations ofleptospirosis.

Leptospirosis is caused by pathogenic leptospires, posing a significant public health problem. Host susceptibility to Leptospira infection is a multifactorial trait, and the host's genetic background can influence both the establishment of infection and the severity of the disease. Complement Factor H (FH) plays a crucial role in the interaction between pathogenic bacteria and the host. Genetic variants in the FH gene CFH have previously been associated with non-infectious diseases. Here, we aimed to analyze the effect of CFH variants on individual susceptibility to leptospirosis and disease severity. To accomplish this, we sequenced CFH exons 7, 9, 21, 22, and 23 in a case/control cohort (184/162) from two endemic leptospirosis areas in Brazil and Argentina. We identified twenty-one single nucleotide variants (SNVs). In the Brazilian cohort, the intronic variant rs34815383 exhibited a higher frequency in patients than in controls, resulting in a significant association with leptospirosis (p = 0.032; OR: 0.32; 95% CI 0.1-1) and also renal disorder (p = 0.001; OR: 5.3; 95%CI 1.8-15.57). This SNV is reported to be a splicing variant, negatively impacting CFH expression, and has previously been associated with Complement-driven renal disease. A second synonymous variant, rs61822181, was significantly less frequent in patients than in controls (p = 0.002; OR: 7.33; 95% CI 1.59-33.7), representing a protective factor against the development of leptospirosis. Our study represents the first documentation of the frequency of CFH SNVs in South America and identifies the variant rs34815383 T > C as a risk factor for leptospirosis and leptospirosis-related renal complications.

Novel Pyrimidine Tethered Benzamide Derivatives as Potential Anticancer Agents: Synthesis, Characterization, Molecular Docking and In vitro Cytotoxicity Evaluation

Current research work presents the synthesis, characterization, molecular docking study and in vitro cytotoxicity evaluation of novel pyrimidine-tethered benzamide derivatives as potential anticancer agents. The synthesis involved multi-steps procedure, including the synthesis of various chalcones (3a-t) with corresponding ketones (1 and 2) and substituted aldehydes (a-j), followed by pyrimidine amines (5a-t) with condensation of chalcones and guanidine and finally pyrimidinyl benzamide derivatives (8a-t) from compounds 5a-t coupling with acid chloride (7) using DIBAL-H. Synthesized compounds characterized by NMR spectroscopy and HRMS. The molecular docking studies were conducted against EGFR (6LUD) and CDK-4 (7SJ3) receptors, revealing distinct binding affinities influenced by substituent groups. Additionally, the cytotoxicity of the synthesized compounds was evaluated using the MTT assay against various cancer cell lines, including A-549 (non-small cell lung cancer), HCT-116 (colorectal cancer), PANC-1 (pancreatic cancer) and HaLa (cervical cancer), along with one normal human embryonic kidney cell line (HEK-293). Among the synthesized compounds, derivatives 8f and 8j exhibited the best anticancer activity against A-549 cells, compounds 8j and 8e showed exceptional potency against HCT-116 cells, compounds 8f and 8j demonstrated high efficacy against PANC-1 cells and compound 8h displayed remarkable potency against HaLa cells. The findings highlight the potential of these pyrimidinyl benzamide derivatives as targeted anticancer agents.

Duplex Kidney with Spontaneous Intraperitoneal Rupture of the Lower Moiety Pyonephrosis: An Extremely Rare Case Report

Introduction: Spontaneous intraperitoneal rupture of pyonephrosis is an extremely rare condition. It leads to generalized peritonitis that mimics an acute abdomen caused by intraperitoneal visceral pathology. In most cases, the underlying cause is obstructive urinary tract stones, followed by pelviureteric junction obstruction, tumors, or connective tissue diseases. We report the first case of spontaneous intraperitoneal fistulization of pyonephrosis from a duplex kidney. Case Presentation: A 56-year-old female presented with an acute abdomen and was diagnosed with a right incomplete duplex kidney and intraperitoneal rupture of lower renal moiety pyonephrosis secondary to an obstructive ureteric calculus located proximal to the confluence of the two ureters. She was successfully managed with initial percutaneous drainage and nephrostomy, followed by a delayed lower moiety nephrectomy. Conclusions: This case highlights the importance of considering renal etiology in the differential diagnosis of generalized peritonitis and underscores the rarity of such occurrences.

Proximal tubule cell maturation rate and function are controlled by PPARα signaling in kidney organoids

Human pluripotent stem cell-derived kidney organoids are expected to be a useful tool for new drug discoveries, however, the immaturation of kidney organoids causes difficulties in recapitulating renal pharmacokinetics using organoids. Here, we performed time-course single-cell RNA sequencing of kidney organoids and revealed cell heterogeneity in the maturation rate of the proximal tubule. An unbiased analysis to identify upstream targets of genes that are expressed differentially between cells with low and high maturation rates revealed a higher activation of PPARα signaling in rapidly maturing cells. Treatment with a combination of a PPARα agonist and an RXRα agonist induced genes related to proximal tubule maturation and increased the capacity for protein uptake as well as the sensitivity to nephrotoxicity by cisplatin. This method to promote the maturation rate of proximal tubule cells has the potential to be utilized in microphysiological systems to recapitulate proximal tubule functions and to screen nephrotoxic drugs.

Post-ischemic inactivation of HIF Prolyl Hydroxylases in endothelium promotes maladaptive kidney repair by inducing glycolysis.

Ischemic acute kidney injury (AKI) is common in hospitalized patients and increases the risk for chronic kidney disease (CKD). Impaired endothelial cell (EC) functions are thought to contribute in AKI to CKD transition, but the underlying mechanisms remain unclear. Here, we identify a critical role for endothelial oxygen sensing prolyl hydroxylase domain (PHD) enzymes 1-3 in regulating post-ischemic kidney repair. In renal endothelium, we observed compartment-specific differences in the expression of the three PHD isoforms in both mice and humans. Post-ischemic concurrent inactivation of endothelial PHD1, PHD2, and PHD3 but not PHD2 alone promoted maladaptive kidney repair characterized by exacerbated tissue injury, fibrosis, and inflammation. Single-cell RNA-seq analysis of the post-ischemic endothelial PHD1, PHD2 and PHD3 deficient (PHDTiEC) kidney revealed an endothelial hypoxia and glycolysis related gene signature, also observed in human kidneys with severe AKI. This metabolic program was coupled to upregulation of the SLC16A3 gene encoding the lactate exporter monocarboxylate transporter 4 (MCT4). Strikingly, treatment with the MCT4 inhibitor syrosingopine restored adaptive kidney repair in PHDTiEC mice. Mechanistically, MCT4 inhibition suppressed pro-inflammatory EC activation reducing monocyte-endothelial cell interaction. Our findings suggest avenues for halting AKI to CKD transition based on selectively targeting the endothelial hypoxia-driven glycolysis/MCT4 axis.

Clinico-Pathological Evaluation of Critically Ailing Dogs with Special Reference to Thrombocytopenia

Background: Thrombocytopenia is the decrease in the number of platelets in blood. In dogs, thrombocytopenia occurs due to a multitude of aetiologies such as haemoprotozoal infections, bacterial infections, viral infections, non-infectious etiologies such as immune dysregulation, neoplasia and even due to miscellaneous conditions. Diagnosis of thrombocytopenia is done mostly by estimation of platelet count in automated haematology analysers and by evaluation of platelet count in blood smears. As clinical research focusing exclusively on canine thrombocytopenia is very limited in India, the present study was carried out to study the aetiology of canine thrombocytopenia and to obtain extensive information regarding qualitative and quantitative changes in platelets in various emergency conditions of dogs. Methods: 450 dogs were diagnosed with thrombocytopenia based on platelet count and a detailed investigation was carried out on the thrombocytopenic dogs to identify the aetiologic agents and to record the quantitative and qualitative changes in platelets. Result: Population incidence of thrombocytopenia in critically ill dog population was recorded as 34.09 per cent. Haemoparasitic infection, renal disorders, hyperthermia and sepsis were recorded as the major causes of thrombocytopenia. Haematobiochemical investigation revealed a significant reduction (p less than 0.01) of hemoglobin, RBC and PCV values and elevation in blood urea nitrogen (BUN), creatinine, alkaline phosphatase (ALP), total bilirubin, direct bilirubin and phosphorus. A mortality of 10.67% (48/450) was recorded in the thrombocytopenic dogs.

Gucy1α1 specifically marks kidney, heart, lung and liver fibroblasts.

Fibrosis is a common outcome of numerous pathologies, including chronic kidney disease (CKD), a progressive renal function deterioration. Current approaches to target activated fibroblasts, key effector contributors to fibrotic tissue remodeling, lack specificity. Here, we report Gucy1α1 as a specific kidney fibroblast marker. Gucy1α1 levels significantly increased over the course of two clinically relevant murine CKD models and directly correlated with established fibrosis markers. Immunofluorescent (IF) imaging showed that Gucy1α1 comprehensively labelled cortical and medullary quiescent and activated fibroblasts in the control kidney and throughout injury progression, respectively. Unlike traditionally used markers platelet derived growth factor receptor beta (Pdgfrβ) and vimentin (Vim), Gucy1α1 did not overlap with off-target populations such as podocytes. Notably, Gucy1α1 labelled kidney fibroblasts in both male and female mice. Furthermore, we observed elevated GUCY1α1 expression in the human fibrotic kidney and lung. Studies in the murine models of cardiac and liver fibrosis revealed Gucy1α1 elevation in activated Pdgfrβ-, Vim- and alpha smooth muscle actin (αSma)-expressing fibroblasts paralleling injury progression and resolution. Overall, we demonstrate Gucy1α1 as an exclusive fibroblast marker in both sexes. Due to its multiorgan translational potential, GUCY1α1 might provide a novel promising strategy to specifically target and mechanistically examine fibroblasts.

Geometrically engineered organoid units and their assembly for pre-construction of organ structures.

Regenerative medicine is moving from the nascent to the transitional stage as researchers are actively engaged in creating mini-organs from pluripotent stem cells to construct artificial models of physiological and pathological conditions. Currently, mini-organs can express higher-order functions, but their size is limited to the order of a few millimeters. Therefore, one of the ultimate goals of regenerative medicine, "organ replication and transplantation with organoid," remains a major obstacle. Three-dimensional (3D) bioprinting technology is expected to be an innovative breakthrough in this field, but various issues have been raised, such as cell damage, versatility of bioink, and printing time. In this study, we established a method for fabricating, connecting, and assembling organoid units of various shapes independent of cell type, extracellular matrix, and adhesive composition (unit construction method). We also fabricated kidney tissue-like structures using three types of parenchymal and interstitial cells that compose the human kidney and obtained findings suggesting the possibility of crosstalk between the units. This study mainly focuses on methods for reproducing the structure of organs, and there are still issues to be addressed in terms of the expression of their higher-order functions. We anticipate that engineering innovation based on this technique will bring us closer to the realization of highly efficient and rapid fabrication of full-scale organoids that can withstand organ transplantation.