Normal Glomeruli Research Articles

Assessment of histopathological effect of factory effluents on the liver and kidney tissues of chickens (Gallus gallus domestica) reared around factory sites in Nnewi Metropolis, Anambra State, Nigeria

Exposure to factory effluents comes with deleterious consequences. The purpose of this study was to assess the histopathological effect of factory effluents on the liver and kidney tissues of chickens (Gallus gallus domestica) reared around factory sites in Nnewi Metropolis, Anambra State, Nigeria. A total of twenty-nine chicks comprising of sixteen chicks exposed to factory sites and thirteen non-exposed chicks were grown to adult birds (chickens) for the study and were allowed to feed from the surrounding homeland of the factories including lead acid battery manufacturing factory (A), metal fabricating factory (B) and metal forging factory (C) respectively until they were aged between four and five months as adult birds (chickens) for the study. The chickens were sacrificed to obtain the liver and kidney tissues for histological analyses. The histopathological evaluation of the effect of the factory effluents on the liver and kidney tissues of birds reared around the four factories (A-D) and control were done by microscopic examination of haematoxylin and eosin (H&E) stained sections. The results obtained in factory A, B and C showed widening of the liver central vein and thickening of the vessels with invasion of the surrounding areas by chronic inflammatory cells as against the controls which showed fairly normal liver tissues with normal liver parenchyma cells with a central vein containing blood cells. Also, the photomicrographs of the kidney tissues of birds reared around factory A, B and C showed evidence of marked glomerulo-nephritis and hypercellularity with evidence of glomerulosclerosis and hyalinization of the glomeruli as against the normal glomeruli and tubules found in the control birds. These results indicate that the environment where these birds are reared is a potential threat to the lives of the birds reared around these factories. Keywords: Factory site, Factory effluents, Liver, Kidney, Chicken (Gallus gallus domestica), Nnewi metropolis.

In vitro and in vivo assessment of banana peel powder as an aflatoxins biosorbent

SURVEY of fungi in raw peanut seeds by washing technique method resulted in isolation of nine fungal species. The aflatoxigenic Aspergillus flavus was the most frequently isolated species in the isolation plates. Molecular identification confirms that it is identical to A. flavus available in database in NCBI GenBank with similarity of 100%. The aflatoxins (AFs) produced in the fermentation medium of A. flavus were extracted and estimated by HPLC to be contain B1, B2, G1, G2 AFs. Banana peel powder, prepared from samples collected from juice markets, proved to be of high adsorption capacity to aflatoxins in concentration depending manner of either peel or aflatoxins.In vivo histopathological studies using 6 albino rats groups, an alteration in rats treated with single aflatoxin concentrations (G2 or G3) such as inflammation, fibrosis, expanded portal tract and degeneration of hepatocytes in liver and expanded glomeruli, vacuolation & fatty degeneration in renal tubules were detected in kidney. However, banana peel treated singly showing normal hepatocytes and central vein in liver as well as normal tubules and glomeruli in kidney. Co-addition of AFs with banana peel powder to rat diet showing significant reduction in the harmful effect of AFs added singly in liver and kidney tissues. This study indicated also that the liver was highly affected by AFs than kidney.

Development and validation of a renal risk score in ANCA-associated glomerulonephritis

Predicting renal outcome in antineutrophil cytoplasmic antibody (ANCA)-associated glomerulonephritis (GN) remains a major challenge. We aimed to identify reliable predictors of end-stage renal disease (ESRD) and to develop and validate a clinicopathologic score to predict renal outcome in ANCA-associated GN. In a prospective training cohort of 115 patients, the percentage of normal glomeruli (without scarring, crescents, or necrosis within the tuft) was the strongest independent predictor of death-censored ESRD. Regression tree analysis identified predictive cutoff values for three parameters: percentage normal glomeruli (N0 >25%, N1 10 to 25%, N2<10%), percentage tubular atrophy and interstitial fibrosis (T0≤25%, T1 >25%), and estimated glomerular filtration rate at the time of diagnosis (G0 >15 ml/min/1.73 m2, G1≤15 ml/min/1.73 m2). Cox regression analysis was used to assign points to each parameter (N1= 4, N2= 6, T1= 2, G1= 3 points), and the resulting risk score was used to classify predicted ESRD risk as low (0), intermediate (2 to 7), or high (8 to 11 points). The risk score accurately predicted ESRD at 36 months in the training cohort (0%, 26%, and 68%, respectively) and in an independent validation cohort of 90 patients (0%, 27%, and 78%, respectively). Here, we propose a clinically applicable renal risk score for ANCA-associated GN that highlights the importance of unaffected glomeruli as a predictor of renal outcome and allows early risk prediction of ESRD.

Detection and identification of potential transglutaminase 2 substrates in the mouse renal glomeruli

The glomerulus primarily comprises mesangial cells, glomerular microvascular endothelial cells, and podocytes. IgA nephropathy is the most common primary glomerulonephritis worldwide and has a risk of progression to end-stage renal disease. IgA nephropathy is characterized by predominant IgA deposition in the glomerular mesangial area, where TG2 is significantly enhanced. Therefore, identification of glomerular TG2 substrates is the first step in elucidating the role of TG2 as a crosslinking enzyme during disease progression. To clarify potential glomerular TG2 substrates, and to establish a procedure for substrate identification, we attempted to identify those molecules using normal mouse glomeruli. Extracts from mouse glomerular and non-glomerular fractions were treated with our established biotin-labeled substrate peptide, which specifically crosslinks to the lysine-donor substrates depending on TG2 activity. Peptide-incorporated proteins were then purified using avidin resin and identified via mass spectrometry. In parallel, we performed the identification using corresponding samples from TG2 knockout mice. Consequently, potential TG2 substrates were separately identified in glomerular and non-glomerular fractions. They were mainly identified as novel TG2 substrates and partly include the well-known substrates. These results potentially provide novel insights into the mechanism underlying IgA nephropathy and may help elucidate the physiological functions of TG2.

Renovascular hypertension masquerading as nephrotic proteinuria in an 10-month-old girl — analysis of errors in diagnostic approach

A 10-month-old girl with nephrotic proteinuria, hypercholesterolaemia, hypoalbuminaemia, hyponatraemia, hypokalaemia, hypocalcaemia, metabolic alkalosis, normal auscultatory blood pressure on multiple occasions, and an enlarged left kidney on ultrasound, was referred to hospital. A left-kidney biopsy showed normal glomeruli with mild mesangial proliferation and the thickening of arteriolar walls. Glucocorticoid therapy was started. A further course was complicated by vomiting, irritation, pulsatile fontanel, and an enlarged ventricular system on ultrasound. A ventriculoperitoneal shunt was inserted but, during anaesthesia, repeated automated, oscillometric measurements revealed blood pressure of 220/120 mmHg. Intravenous antihypertensive treatment lowered the blood pressure to 110–120/70 mmHg, with a decrease in proteinuria. A CT abdominal angiography revealed left-renal-artery stenosis, a reduced right kidney and an enlarged left kidney, with foci of a post-infarct area in the lower pole. The patient was referred to a tertiary-care hospital. Renal scintigraphy showed that the right-kidney function had decreased to 6%. A right nephrectomy was performed but, due to persistent arterial hypertension, oral antihypertensive treatment was intensified. Microscopic analysis of the removed right kidney revealed relatively normal glomeruli and fibromuscular dysplasia in the right-renal-artery specimen. Oral antihypertensive therapy based on an angiotensin-converting enzyme inhibitor and a calcium-channel blocker continued after surgery allowed the attaining of normotension, the gradual normalisation of proteinuria and metabolic abnormalities, and an improvement in the left-kidney function. This case shows that auscultatory blood-pressure measurements in small children and incorrect interpretation of the obtained test results can lead to the erroneous misdiagnosis of hypertensive crisis, with dramatic consequences.

Expression of Cathepsin L and Its Intrinsic Inhibitors in Glomeruli of Rats With Puromycin Aminonucleoside Nephrosis.

Cathepsin L, a lysosomal cysteine proteinase, may have a key role in various biological and disease processes by intracellular and extracellular degradation of proteins. We examined the levels of cathepsin L and its intrinsic inhibitors in glomeruli of rats with puromycin aminonucleoside (PAN) nephrosis. In contrast to the weak levels of cathepsin L in normal glomeruli, on days 4 and 8, strong immunostaining was detected in almost all podocytes when proteinuria and pathological changes of the podocytes developed. Cathepsin L was reduced after day 28, but remained in a focal and segmental manner. Cystatin β, an intracellular inhibitor, was not detected in podocytes. However, cystatin C, an extracellular inhibitor, was detected in podocytes after day 4, coincident with cathepsin L. Cystatin C levels were gradually reduced but sustained in many podocytes on day 28, while cystatin C was not detected in podocytes sustained cathepsin L. These results demonstrated that cathepsin L levels are not always accompanied by the levels of its inhibitors in podocytes of PAN nephrosis, suggesting a potential role of cathepsin L in podocyte injury, which is a critical process for the development and progression of tuft adhesion and sclerosis.

An evaluation of the 2010 histopathological classification of anti-neutrophil cytoplasmic antibody (ANCA)-associated glomerulonephritis: a Bayesian network meta-analysis

In 2010, a new histopathological classification of ANCA-associated glomerulonephritis (ANCA-GN) based on four categories (focal, crescentic, mixed, and sclerotic) was proposed to predict renal outcome. However, this classification was the source of much debate in different populations. We aimed to evaluate this classification scheme in terms of renal survival using Bayesian network meta-analysis. We searched Pubmed, Embase, and Medline for articles published between 1 October 2010 and 9 August 2017 that assessed the predictive value of this classification. We pooled hazard ratios (HRs) for end-stage kidney disease (ESRD) among the four categories using a Bayesian random-effects model. Clinical factors that could potentially influence renal outcome among the different trials were explored by meta-regression. Sixteen case-controlled studies, including a total of 1945 patients with 1695 endpoint events, were included. Compared with sclerotic, the HRs for ESRD were 0.45 [95% confidence interval (CI) 0.26-0.79] for crescentic, 0.34 (0.22-0.51) for mixed, and 0.24 (0.12-0.51) for focal. Pooled results showed no obvious difference between the crescentic and the mixed sub-groups (HR 1.35; 95% CI 0.90-2.0). Baseline eGFR (P = 0.002) and the ANCA serology (P = 0.029) were associated with renal survival. The 2010 ANCA-GN classification and the extent of ANCA serology and baseline eGFR were shown to be significant predictors of renal outcome, although there was no significant prognostic difference between crescentic and mixed. The ANCA-GN classification scheme should, therefore, be optimized by combining with other established parameters, such as tubular atrophy and the proportion of normal glomeruli.

Deep Learning Global Glomerulosclerosis in Transplant Kidney Frozen Sections.

Transplantable kidneys are in very limited supply. Accurate viability assessment prior to transplantation could minimize organ discard. Rapid and accurate evaluation of intra-operative donor kidney biopsies is essential for determining which kidneys are eligible for transplantation. The criterion for accepting or rejecting donor kidneys relies heavily on pathologist determination of the percent of glomeruli (determined from a frozen section) that are normal and sclerotic. This percentage is a critical measurement that correlates with transplant outcome. Inter- and intra-observer variability in donor biopsy evaluation is, however, significant. An automated method for determination of percent global glomerulosclerosis could prove useful in decreasing evaluation variability, increasing throughput, and easing the burden on pathologists. Here, we describe the development of a deep learning model that identifies and classifies non-sclerosed and sclerosed glomeruli in whole-slide images of donor kidney frozen section biopsies. This model extends a convolutional neural network (CNN) pre-trained on a large database of digital images. The extended model, when trained on just 48 whole slide images, exhibits slide-level evaluation performance on par with expert renal pathologists. Encouragingly, the model's performance is robust to slide preparation artifacts associated with frozen section preparation. The model substantially outperforms a model trained on image patches of isolated glomeruli, in terms of both accuracy and speed. The methodology overcomes the technical challenge of applying a pretrained CNN bottleneck model to whole-slide image classification. The traditional patch-based approach, while exhibiting deceptively good performance classifying isolated patches, does not translate successfully to whole-slide image segmentation in this setting. As the first model reported that identifies and classifies normal and sclerotic glomeruli in frozen kidney sections, and thus the first model reported in the literature relevant to kidney transplantation, it may become an essential part of donor kidney biopsy evaluation in the clinical setting.

Novel hemodynamic structures in the human glomerulus.

To investigate human glomerular structure under conditions of physiological perfusion, we have analyzed fresh and perfusion-fixed normal human glomeruli at physiological hydrostatic and oncotic pressures using serial resin section reconstruction, confocal, multiphoton, and electron microscope imaging. Afferent and efferent arterioles (21.5 ± 1.2 µm and 15.9 ± 1.2 µm diameter), recognized from vascular origins, lead into previously undescribed wider regions (43.2 ± 2.8 µm and 38.4 ± 4.9 µm diameter) we have termed vascular chambers (VCs) embedded in the mesangium of the vascular pole. Afferent VC (AVC) volume was 1.6-fold greater than efferent VC (EVC) volume. From the AVC, long nonbranching high-capacity conduit vessels (n = 7) (Con; 15.9 ± 0.7 µm diameter) led to the glomerular edge, where branching was more frequent. Conduit vessels have fewer podocytes than filtration capillaries. VCs were confirmed in fixed and unfixed specimens with a layer of banded collagen identified in AVC walls by multiphoton and electron microscopy. Thirteen highly branched efferent first-order vessels (E1; 9.9 ± 0.4 µm diameter) converge on the EVC, draining into the efferent arteriole (15.9 ± 1.2 µm diameter). Banded collagen was scarce around EVCs. This previously undescribed branching topology does not conform to the branching of minimum energy expenditure (Murray’s law), suggesting that even distribution of pressure/flow to the filtration capillaries is more important than maintaining the minimum work required for blood flow. We propose that AVCs act as plenum manifolds possibly aided by vortical flow in distributing and balancing blood flow/pressure to conduit vessels supplying glomerular lobules. These major adaptations to glomerular capillary structure could regulate hemodynamic pressure and flow in human glomerular capillaries.

Prognostic Factors in Anti-Neutrophil Cytoplasmic Antibody-Associated Glomerulonephritis with Severe Glomerular Sclerosis: A National Registry-Based Cohort Study.

Background Classification of patients with anti-neutrophil cytoplasmic antibody-associated glomerulonephritis (ANCA-GN) into histological classes is useful for predicting a patient's risk of progression to end-stage renal disease (ESRD). However, even in the worst prognostic group, the 5-year end-stage renal disease-free survival rate is as high as 50%. Objectives To investigate those prognostic factors indicative of progression to ESRD in patients with ANCA-GN and sclerosing histology. Methods Patients from the Norwegian Kidney Biopsy Registry between 1991 and 2012 who had biopsy verified pauci-immune glomerulonephritis, positive ANCA serology, and sclerosing histology were included. Cases with ESRD during follow-up were identified via linkage with the Norwegian Renal Registry. Potential prognostic factors with relevant cut-offs were compared in patients with and without progression to ESRD during follow-up. Results Of 23 included patients, 10 progressed to ESRD. ESRD patients had a lower initial estimated glomerular filtration rate (eGFR; 21 versus 52 ml/min/1.73 m2) and a lower percentage of normal glomeruli (4% versus 15%). Five-year risks of ESRD with eGFR >15 versus ≤15 ml/min/1.73 m2 were 77% and 15%, with percentage normal glomeruli >10% versus ≤10%, 83% and 39%. Conclusions eGFR and percentage of normal glomeruli are strong risk factors for ESRD in ANCA-GN with sclerosing histology.

Gambaran Histologi Ginjal Tikus Wistar Yang Terpapar MSG Setelah Perlakuan Diberikan Jus Tomat Dan Diberhentikan Perlakuan Saja

MSG causes many side effects on the body, but MSG has long been used as a food flavor that can bring tastes (umami) and play a role in strengthening the taste. The chemical structure of MSG is no different from Glutamic Acid (glutamate), it is one of the 20 amino acids that make up proteins in the body. So the use of MSG needs to be discontinued to prevent kidney damage due to continued consumption of MSG, two research were conducted to determine the effect of discharging of MSG consumption by being given tomato juice (1) and discharged without treatment (2), the second study was seen from the histology picture of rats wistar mouse.Methods were experimental with post test only control group design. The research (1) used 15 wistar rats divided into three groups: group I without treatment (pellet AD II and drinking water), group II was given MSG for 14 days and group III was given tomato juice and MSG for 14 days. The study (2) used 27 rats divided by 9 groups. The sample is selected by simple random sampling method. The mice were then gradually turned off on the 29th, 43rd and 57th days. There was no significant difference in mean number of normal and damaged proximal tubules in all treatment groups. The renal histologic features in the treatment group I (MSG) and the treatment group II (MSG and tomato juice) showed normal glomeruli, tubular epithelial tubules, and lumen tubular narrowing whereas in mice discharged without treatment there was no apparent difference between the two MSG Giving groups damage to proximal tubules and renal corpusculum and regeneration after 14 days of discontinuation of MSG. Conclusions from these two experiments occurred damage to the renal tubules Keywords: MSG, Kidney, Wistar Rat

Prognosis and its risk factors in ANCA-associated glomerulonephritis patients treated with initial renal replacement therapy

Objective: To explore the prognosis and its risk factors in anti-neutrophil cytoplasmic antibodies (ANCA)-associated glomerulonephritis (AAGN) patients who needed initial renal replacement therapy (RRT). Methods: One hundred patients [54 females, 46 males, with a median age of 54(41, 60) years] with biopsy-proven AAGN and requiring initial RRT between January 1996 and December 2016 in Nanjing Jinling Hospital were included. Intensive immunotherapy indicated that the patients received corticosteroids in combination with cyclophosphamide or mycophenolate mofetil, or immunoadsorption (IA) or double filtration plasmapheresis (DFPP). The clinical and histological risk factors for renal survival were analyzed. Results: Forty-one patients were free of RRT after a median time of 1 (0.5, 2) month treatment (dialysis-independent group), and the remaining 59 patients were on maintenance dialysis (dialysis-dependent group). The multivariate logistic analysis revealed that the proportion of normal glomeruli <8% (OR=5.95, P=0.002) and global sclerotic glomeruli ≥50% (OR=4.87, P=0.003), and not receiving intensive immunotherapy (OR=7.81, P=0.004) were the risk factors for the renal recovery in these patients. During a median follow-up time of 22 (10, 50) months, 15 patients(36.6%) in the dialysis-independent group progressed into maintenance dialysis, and the 1 and 3 year renal survival rate were 86% and 60%, respectively. During a median follow-up time of 6 (2, 24) months, 12 (12%) patients died, among whom four patients died of therapy. The multivariate Cox regression analysis revealed that IA/DFPP treatment (HR=10.85, P=0.034) and low albumin level (HR=1.26, P=0.009) significantly associated with a higher risk of therapy-related death. Conclusions: The renal recovery rate in AAGN patients with initial RRT was low. The proportion of normal and global sclerotic glomeruli, receiving intensive immunotherapy or not were associated with renal outcome, and IA/DFPP treatment as well as lower albumin level were independently associated with therapy-related death.

Investigation into the underlying molecular mechanisms of hypertensive nephrosclerosis using bioinformatics analyses.

Hypertensive nephrosclerosis (HNS) is a major risk factor for end-stage renal disease. However, the underlying pathogenesis of HNS remains to be fully determined. The gene expression profile of GSE20602, which consists of 14 glomeruli samples from patients with HNS and 4 normal glomeruli control samples, was obtained from the Gene Expression Omnibus database. Gene ontology (GO) and pathway enrichment analyses were performed in order to investigate the functions and pathways of differentially expressed genes (DEGs). Pathway relation and co-expression networks were constructed in order to identify key genes and signaling pathways involved in HNS. In total, 483 DEGs were identified to be associated with HNS, including 302 upregulated genes and 181 downregulated genes. Furthermore, GO analysis revealed that DEGs were significantly enriched in the small molecule metabolic process. In addition, pathway analysis also revealed that DEGs were predominantly involved in metabolic pathways. The tricarboxylic acid (TCA) cycle was identified as the hub pathway in the pathway relation network, whereas the sorbitol dehydrogenase (SORD) and cubulin (CUBN) genes were revealed to be the hub genes in the co-expression network. The present study revealed that the SORD, CUBN and albumin genes as well as the TCA cycle and metabolic pathways are involved in the pathogenesis of HNS. The results of the present study may contribute to the determination of the molecular mechanisms underlying HNS, and provide insight into the exploration of novel targets for the diagnosis and treatment of HNS.

Predicting Outcome in Patients with Anti-GBM Glomerulonephritis.

Large studies on long-term kidney outcome in patients with anti-glomerular basement membrane (anti-GBM) GN are lacking. This study aimed to identify clinical and histopathologic parameters that predict kidney outcome in these patients. This retrospective analysis included a total of 123 patients with anti-GBM GN between 1986 and 2015 from six centers worldwide. Their kidney biopsy samples were classified according to the histopathologic classification for ANCA-associated GN. Clinical data such as details of treatment were retrieved from clinical records. The primary outcome parameter was the occurrence of ESRD. Kidney survival was analyzed using the log-rank test and Cox regression analyses. The 5-year kidney survival rate was 34%, with an improved rate observed among patients diagnosed after 2007 (P=0.01). In patients with anti-GBM GN, histopathologic class and kidney survival were associated (P<0.001). Only one of 15 patients with a focal class biopsy sample (≥50% normal glomeruli) developed ESRD. Patients with a sclerotic class biopsy sample (≥50% globally sclerotic glomeruli) and patients with 100% cellular crescents did not recover from dialysis dependency at presentation. In multivariable analysis, dialysis dependency at presentation (hazard ratio [HR], 3.17; 95% confidence interval [95% CI], 1.59 to 6.32), percentage of normal glomeruli (HR, 0.97; 95% CI, 0.95 to 0.99), and extent of interstitial infiltrate (HR, 2.02; 95% CI, 1.17 to 3.50) were predictors of ESRD during follow-up. Dialysis dependency, low percentage of normal glomeruli, and large extent of interstitial infiltrate are associated with poor kidney outcome in anti-GBM GN. Kidney outcome has improved during recent years; the success rate doubled after 2007. This article contains a podcast at https://www.asn-online.org/media/podcast/CJASN/2017_11_21_CJASNPodcast_18_1_v.mp3.

HEPATO-NEPHROPROTECTIVE ACTIVITY OF NIGELLA SATIVA OIL ON PARACETAMOL-INDUCED NEW ZEALAND RABBITS (ORYCTOLAGUS CUNICULUS)

Objective: The objective of this work was to investigate the hepato-nephroprotective activity of Nigella sativa (Ranunculaceae) oil on paracetamol-induced New Zealand rabbits (Oryctolagus cuniculus).Methods: Hepato-nephroprotective activity of Nigella sativa oil was demonstrated on six groups of paracetamol-induced New Zealand rabbits (Oryctolagus cuniculus) aged 3-4 mo, three in each group (2 males, 1 female). Group I was normal control (water 1.0 ml/kg of body weight per oral), group II was negative control (water 1.0 ml/kg of body weight per oral), group III was positive control (silymarin 100 mg/kg of body weight per oral), group IV-VI were treated with Nigella sativa oil (NSO) dose of 0.5 mg/kg of body weight, 1.0 mg/kg of body weight, and 2.0 mg/kg of body weight per oral, respectively, for 15 d. At the 16th day, rabbits in group II-VI were induced with paracetamol at a dose of 600 mg/kg of body weight per oral. At the 23rd day the animals were measured for their clinical biochemistry parameters and histological examination.Results: Paracetamol administration dose of 600 mg/kg of BW resulted in the elevation of serum glutamic-oxaloacetic transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT), and ureum-N levels of the animals, particularly in group II which was treated only with paracetamol. Normal histology of the liver defines the clear shape of the terminal hepatic venule (THV)/central vein (CV) and sinusoids, whereas that of the kidney defines clear shape of the Bowman capsule and glomerulus shape. Qualitative histological examination of the liver showed that the THV/CV in all groups was normal, however in the paracetamol-treated group, the sinusoids were dilated, necrosis and mass apoptosis were detected. Dilated sinusoids were observed in the silymarin group and in the lower and medium doses of NSO groups. In the highest dose of NSO group the THV/CV and sinusoids were normal, but a local apoptosis and fat degeneration were detected. Qualitative histological examination of the kidney indicated that there was no abnormality of the glomerulus shape, however, mass apoptosis and local necrosis of the kidney were found in the paracetamol-treated group and the silymarin-treated group. The lowest dose of the NSO-treated group showed a normal shape of glomerulus and Bowman capsule, normal apoptosis. No necrosis was observed in the rabbit’s kidney. Higher doses of NSO groups indicated a normal glomerulus shape and Bowman capsule, mass apoptosis and local necrosis.Conclusion: In this study, Nigella sativa oil could maintain the normality of the THV/CV and sinusoids in the liver of paracetamol-induced New Zealand rabbits (Oryctolagus cuniculus). Normal glomerulus shape and Bowman capsule were also confirmed in the kidney of paracetamol-induced animals.

Complement regulatory proteins in kidneys of patients with anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis.

The complement system activation is involved in the development of anti-neutrophil cytoplasmic antibody-associated vasculitis (AAV). The study aimed to investigate the expression of complement regulatory proteins (CRPs) CD46, CD55 and CD59 in kidneys of 51 AVV patients. The expression of CD46, CD55 and CD59 in kidneys was detected by immunohistochemistry and double immunofluorescence staining. The immunohistochemical examination revealed that expression of the three CRPs could be detected in the glomeruli and tubules of both AAV patients and normal controls. The expression levels of the three CRPs in glomeruli of patients with AAV were significantly lower than those of normal controls. The scores of CD46 and CD55 expression in the tubules of AAV patients were significantly lower than those of normal controls, while there was no significant difference between the scores of CD59 expression in tubules of AAV patients and those of normal controls. Among AAV patients, the expression level of CD46 in glomeruli correlated inversely with the proportion of normal glomeruli, while it correlated with tubular atrophy in renal interstitium (r=-0·305, P=0·026; r=0·330, P=0·023, respectively). The expression levels of CD55 and CD59 in glomeruli correlated with the proportion of total crescents (r=0·384, P=0·006; r=0·351, P=0·011, respectively). Double immunofluorescence staining indicated that all three CRPs were expressed on endothelial cells, podocytes and mesangial cells in glomeruli. The expression levels of the three CRPs were dysregulated in kidneys of patients with AAV. The expression levels of CD46, CD55 and CD59 were associated with the severity of renal injury of AAV patients.

Human umbilical cord Wharton jelly cells promote extra-pancreatic insulin formation and repair of renal damage in STZ-induced diabetic mice

BackgroundWe evaluated the therapeutic effect and fate of high doses of human umbilical cord Wharton jelly cells (hUCWJCs) after IP administration to streptozotocin (STZ)-induced diabetic mice.MethodsType 1 diabetes (T1D) was induced in Kunming mice via IP injection of STZ. hUCWJCs were labeled with 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate (DiI). Diabetic animals with sustained hyperglycemia for at least 2 weeks were administered 1 × 107 Dil-hUCWJCs via intraperitoneal injection. Insulin, glucagon and PDX-1 were detected by immunofluorescence with confocal microscopy. Serum mouse and human C-peptide was assayed in blood collected via intracardiac puncture. Specific β-cell differentiation markers and human DNA were assessed using qPCR performed with 200 ng of target DNA.ResultshUCWJCs migrated to the STZ-damaged organs and contributed to lower blood glucose levels in 30% of the treated mice. Confocal microscopy revealed the presence of resident insulin-positive cells in the liver and kidneys. hUCWJC-treated mice with restored hyperglycemia also showed increased serum mouse C-peptide levels. The qPCR results, particularly in the liver, revealed that after transplantation hUCWJCs upregulated genes of endocrine precursors but failed to express endocrine stage markers. Mice with restored hyperglycemia had reduced urinary volume and lacked glomerular hypertrophy, exhibiting a morphology resembling that of normal glomeruli. Moreover, we also verified that one of the possible mechanisms by which hUCWJCs exert immunosuppressive effects is through down-regulation of the cell surface receptor HLA-1.ConclusionsWe confirmed the potential of IP administration of hUCWJCs and the capability of these cells to migrate to damaged tissues and promote insulin secretion from non-pancreatic local cells and to improve renal damage. These findings confer unique therapeutic properties to hUCWJCs, suggesting a promising future in the treatment of diabetes mellitus.

Abolishment of proximal tubule albumin endocytosis does not affect plasma albumin during nephrotic syndrome in mice

The megalin/cubilin receptor complex is required for proximal tubular endocytosis and degradation of filtered albumin. An additional high-capacity retrieval pathway of intact albumin for the recovery of large amounts of filtered albumin has been proposed, possibly involving cooperation between megalin/cubilin and the neonatal Fc receptor. To clarify the potential role of such a pathway, we examined the effects of megalin/cubilin gene inactivation on tubular albumin uptake and plasma albumin levels in nephrotic, podocin knockout mice. Immunofluorescence microscopy of megalin/cubilin/podocin knockout mouse kidneys demonstrated abolishment of proximal tubule albumin uptake, in contrast to the excessive albumin accumulation observed in podocin knockout mice compared to controls. Correspondingly, urinary albumin excretion was increased 1.4 fold in megalin/cubilin/podocin compared to podocin knockout mice (albumin/creatinine: 226 vs. 157 mg/mg). However, no difference in plasma albumin levels was observed between megalin/cubilin/podocin and podocin knockout mice, as both were reduced to approximately 40% of controls. There were no differences in liver albumin synthesis by mRNA levels and protein abundance. Thus, megalin/cubilin knockout efficiently blocks proximal tubular albumin uptake in nephrotic mice but plasma albumin levels did not differ as a result of megalin/cubilin-deficiency, suggesting no significance of the megalin/cubilin-pathway for albumin homeostasis by retrieval of intact albumin.

Urinary protein profiles in ketorolac-associated acute kidney injury in patients undergoing orthopedic day surgery.

BackgroundParenteral administration of ketorolac is very effective in controlling postoperative pain for orthopedic surgery. Ketorolac can induce clinically relevant renal alterations in elderly patients, whereas its short course is considered safe for young adults with normal preoperative renal function. In this study, of a cohort of young adults undergoing elective orthopedic day surgery, we sought cases complicated by readmission due to acute kidney injury (AKI).Patients and methodsAmong 1397 young adults, aged 18–32 years who were admitted to undergo orthopedic day surgery from 2013 to 2015, four patients (0.29%, three males/one female) treated in postprocedure with ketorolac (from 60 to 90 mg/day for 1–2 days) were readmitted for suspected severe AKI. We evaluated functional outcome, urinary protein profiles and kidney biopsy (1 patient).ResultsAfter day surgery discharge, they experienced gastrointestinal disturbances, flank pain and fever. Readmitted on post-surgery days 3–4, they presented with oliguric AKI (creatinine range 158.4–466.4 µmol/L) and frank proteinuria (albumin range 2.1–6.0 g/L). Urine protein profiles demonstrated a nonselective glomerular proteinuria, with a significant 9.4-fold increase in glomerular/tubular index on day 6. Kidney biopsy on day 19 showed normal glomeruli and minimal tubular alterations and negative immunofluorescence. All patients recovered their renal function, and after 20 days proteinuria disappeared.ConclusionAKI can ensue even in young adults who have undergone a short course of ketorolac, when they suffered from relative dehydration, abdominal disturbances, flank pain and oliguria after discharge. Urine findings were characterized by a marked nonselective glomerular proteinuria disappearing in 2–3 weeks.

Relevant genetic polymorphisms and kidney expression of Toll-like receptor (TLR)-5 and TLR-9 in lupus nephritis.

Toll-like receptor (TLR) genetic polymorphisms may modify their expression causing inflammatory disorders and influencing both susceptibility and severity of lupus erythematosus. We aim to determine whether TLR-5 and TLR-9 gene polymorphisms are implicated in the susceptibility to systemic lupus erythematosus (SLE) and lupus nephritis (LN) and to evaluate their expressions and distributions in renal LN patients' biopsies. The frequencies of two SNP in the TLR-9 gene and one in the TLR-5 gene was examined in 106 SLE patients (among them 37 LN patients) and in 200 matched controls by polymerase chain reaction-restriction fragment-length polymorphisms (PCR-RFLP) analysis. TLR-9 and TLR-5 expressions were assessed by reverse transcription (RT)-PCR and immunohistochemistry carried on LN renal biopsies compared to healthy renal tissue. A significant genotypic and allelic association was revealed between TLR-9-rs352140 and both SLE and LN (P < 0·05). The TLR-9 transcript level was significantly higher in LN biopsies compared to control (P < 0·05). This increase was observed histochemically in the tubulointerstitial compartment. TLR-9 was detectable in LN glomeruli patients but not in normal control glomeruli. No allelic nor genotype association was found with TLR-5-rs5744168 in SLE. but the T allele and the TT genotype were raised significantly in the LN group (P < 0·05). A significant increase in TLR-5 gene expression in LN biopsies, which contrasted with normal kidneys (P < 0·05), was confirmed by an intense and diffuse staining for TLR-5 only in LN tubules (P < 0·05). Our data show that TLR-5 and TLR-9 are susceptible genes to LN and that their expression is dysregulated in LN patients' kidneys, supporting a role of these mediators in the pathogenesis of LN.