Kidney Glomeruli Research Articles

IL-1 receptor signaling in podocytes limits susceptibility to glomerular damage.

Interleukin (IL)-1 receptor type 1 (IL-1R1) activation triggers a proinflammatory signaling cascade that can exacerbate kidney injury. However, the functions of podocyte IL-1R1 in glomerular disease remain unclear. To study the role of IL-1R1 signaling in podocytes, we selectively ablated podocyte IL-1R1 in mice (PKO mice). We then subjected PKO mice and wild-type controls to two glomerular injury models: nephrotoxic serum (NTS)- and adriamycin-induced nephropathy. Surprisingly, we found that IL-1R1 activation in podocytes limited albuminuria and podocyte injury during NTS- and adriamycin-induced nephropathy. Moreover, deletion of IL-1R1 in podocytes drove podocyte apoptosis and glomerular injury through diminishing Akt activation. Activation of Akt signaling abrogated the differences in albuminuria and podocyte injury between wild-type and PKO mice during NTS. Thus, IL-1R1 signaling in podocytes limits susceptibility to glomerular injury via an Akt-dependent signaling pathway. These data identify an unexpected protective role for IL-1R1 signaling in podocytes in the pathogenesis of glomerular disease.NEW & NOTEWORTHY The present study establishes that activation of the receptor for interleukin-1 limits susceptibility to damage to the kidney glomerulus in preclinical mouse models by stimulating Akt signaling cascades inside the podocyte.

Chronic intracellular hypoxia as a clustering and stratifying factor for clinical severity grade in nephrotic syndrome in children

Abstract. Nephrotic syndrome is the most common glomerular kidney disease in childhood. It is known that chronic hypoxia is a severe disorder and potent factor of kidney damage. The limited success of existing therapeutic strategies in slowing the progression of chronic kidney disease requires the study of new ways to assess and interpret the levels of chronic intracellular hypoxia concerning basic clinical data, grades of NS activity in children, type of therapeutic response.
 The study aimed to investigate the state of transcription factor and marker of intracellular hypoxia HIF-1alfa in children with different degrees of change in basic clinical and laboratory parameters; to evaluate HIF-1alfa as a possible factor of stratification of activity grade of nephrotic syndrome.
 Methods. This case-control study was carried out in the duration from June 2018 to August 2020. The study was conducted on 35 selected patients with NS collected from the nephrology department, Pediatric Clinical Hospital №7 (Kyiv, Ukraine). Plasma samples were used to measure marker intracellular hypoxia HIF-1alfa. ANOVA followed by the post hoc Kruskal-Wallis test for multiple comparisons was used to test the significance of differences. GraphPad Prism 9.0 Software for Windows and Statistica 10.0 software used. P values <0,05 considered statistically significant.
 Results. Three groups of children with different activity grades were stratified on basis of indicators of proteinuria levels, total blood protein, blood alpha2-globulin levels, serum cholesterol levels, and edema. 1st-grade group found to have a mild increase of HIF-1alfa up to 185-195 a.u. proteinuria 3,5-5,5 g/24 h, total blood protein 47-53 g/L, alfa2-globulins level in blood 20-23 g/L, serum cholesterol level 6-8,5 mMol/L, edema - 1-1.6 points. 2nd grade group found to have moderate increase of HIF-1alfa up to 195,1-205 a.u., proteinuria 5,51-8,5 g/24 h, total blood protein 46,9-40 g/L, alfa2-globulins level in blood 23,1-27 g/L, serum cholesterol level 8.51-10,5 mMol/L, edema 1.61-2.2 points. 3rd-grade group found to have pronounced increase of HIF-1alfa up to 205,1-220 a.u., proteinuria 8,51-14 g/24 h, total blood protein 39,9-32 g/L, alfa2-globulins level in blood 27,1-30 g/L, serum cholesterol level 10.51-13.5 mMol/L, edema 2.21-3 points. Higher HIF-1alfa level appears in children with NS and frequent relapses as compared to the group with rare relapses.
 Conclusion. Thus, the increase of HIF-1 alpha to the level of 185-205 a.u., which corresponds to the I-II degree of activity in children with NS can be used as a starting point and therapeutic window for specific anti-hypoxic and antioxidant interventions. Determination of HIF-1 alpha levels in children with NS can be used as a factor for stratification of the activity grade.

Bioengineering Strategies to Develop Podocyte Culture Systems.

Unraveling the complex behavior of healthy and disease podocytes by analyzing the changes in their unique arrangement of foot processes, slit diaphragm, and the three-dimensional (3D) morphology is a long-standing goal in kidney-glomerular research. The complexities surrounding the podocytes' accessibility in animal models and growing evidence of differences between humans and animal systems have compelled researchers to look for alternate approaches to study podocyte behaviors. With the advent of bioengineered models, an increasingly powerful and diverse set of tools is available to develop novel podocyte culture systems. This review discusses the pertinence of various culture models of podocytes to study podocyte mechanisms in both normal physiology and disease conditions. While no one in vitro system comprehensively recapitulates podocytes' in vivo architecture, we emphasize how the existing systems can be exploited to answer targeted questions on podocyte structure and function. We highlight the distinct advantages and limitations of using these models to study podocyte behaviors and screen therapeutics. Finally, we discuss various considerations and potential engineering strategies for developing next-generation complex 3D culture models for studying podocyte behaviors in vitro. Impact Statement In various glomerular kidney diseases, there are numerous alterations in podocyte structure and function. Yet, many of these disease events and the required targeted therapies remain unknown, resulting in nonspecific treatments. The scientific and clinical communities actively search for new modes to develop structurally and functionally relevant podocyte culture systems to gain insights into various diseases and develop therapeutics. Current in vitro systems help in some ways but are not sufficient. A deeper understanding of these previous approaches is essential to advance the field, and importantly, bioengineering strategies can contribute a unique toolbox to establish next-generation podocyte systems.

The morphology of the afferent and efferent domain of the sheep glomerulus.

It is important to know the morphology of the glomerulus in order to explain kidney infiltration. The present study aims to research the morphology of afferent and efferent domains of sheep kidney glomeruli. In this study, 2000 glomeruli from 20 kidneys of Akkaraman sheep were examined using the polyester resin method. It was found that the glomeruli of sheep kidney usually had an afferent arteriole as well as an efferent arteriole. Besides, it was also found that five glomeruli had two efferent arterioles. It is known that the afferent domain constitutes the largest part of the glomerulus. In two of the glomeruli that we examined, the afferent domain formed the 1/2 of the glomeruli wherein the other two glomeruli afferent domain formed the 3/4. It is known that there are many anastomoses between the afferent and efferent domain of capillaries. However, it is not well-explained how anastomosis is created between the afferent and efferent domains. In our study, it was identified that those anastomoses were not inside the lobes but between the surrounding capillaries.

GlomSpheres as a 3D co-culture spheroid model of the kidney glomerulus for rapid drug-screening

The glomerulus is the filtration unit of the kidney. Injury to any component of this specialised structure leads to impaired filtration and eventually fibrosis and chronic kidney disease. Current two and three dimensional (2D and 3D) models that attempt to recreate structure and interplay between glomerular cells are imperfect. Most 2D models are simplistic and unrepresentative, and 3D organoid approaches are currently difficult to reproduce at scale and do not fit well with current industrial drug-screening approaches. Here we report a rapidly generated and highly reproducible 3D co-culture spheroid model (GlomSpheres), better demonstrating the specialised physical and molecular structure of a glomerulus. Co-cultured using a magnetic spheroid formation approach, conditionally immortalised (CI) human podocytes and glomerular endothelial cells (GEnCs) deposited mature, organized isoforms of collagen IV and Laminin. We demonstrate a dramatic upregulation of key podocyte (podocin, nephrin and podocalyxin) and GEnC (pecam-1) markers. Electron microscopy revealed podocyte foot process interdigitation and endothelial vessel formation. Incubation with pro-fibrotic agents (TGF-β1, Adriamycin) induced extracellular matrix (ECM) dysregulation and podocyte loss, which were attenuated by the anti-fibrotic agent Nintedanib. Incubation with plasma from patients with kidney disease induced acute podocyte loss and ECM dysregulation relative to patient matched remission plasma, and Nintedanib reduced podocyte loss. Finally, we developed a rapid imaging approach to demonstrate the model’s usefulness in higher throughput pharmaceutical screening. GlomSpheres therefore represent a robust, scalable, replacement for 2D in vitro glomerular disease models.

The analgesic activities of total alkaloids of the ethnic medicine Cynanchum komarovii Al. Iljinski

Ethnopharmacological relevanceCynanchum komarovii Al. Iljinski is a ethnomedicinal herb and this ethno-medicine is used mainly to treat arthritis, toothache, reducing phlegm, relieving cough. Total alkaloids of Cynanchum komarovii Al. Iljinski (TACKI) is the main active compound of Cynanchum komarovii Al. Iljinski. Previous investigations have revealed that TACKI can significantly inhibit rat foot swelling caused by carrageenan; it has a significant inhibitory effect on granulation tissue proliferation. Pharmacology study showed that Cynanchum komarovii Al. Iljinski has analgesia, anti-inflammatory, antibacterial, anti-tumor, relieving cough and relieving asthma. However, there is no any investigation on the mechanism of analgesia and anti-inflammation. Aim of the studyTo clarify the analgesic effect and material basis of Cynanchum komarovii Al. Iljinski, determine the analgesic effect of TACKI, and provide experimental data support for its traditional application in the treatment of various pains. Materials and methodsTACKI were prepared by the traditional acid extraction and alkaline precipitation method, and TACKI was analyzed through classic animal models of acute antinociceptive animal models and chronic antinociceptive. Evaluation of analgesic effects, and preliminary discussion of the mechanism of its analgesic effects were performed in this work. ResultsAcute toxicity experiments showed that the LD50 of TACKI mice was 2960.88 mg/kg, and symptoms of poisoning appeared. Patholog of liver and kidney studies have shown that TACKI reduces eosinophils and increases basophils in kidney glomeruli. In the study of analgesic effects, TACKI had analgesic activity through the PWL, formalin test, and acetic acid writhing test. In the chronic inflammatory antinociceptive study, the latency of the withdrawal reflex in the TACKI group was prolonged, and the mechanical withdrawal reflex threshold was significantly increased. The protein expression of NMDA, GFAP and Iba-1 in rat brain tissue can be reduced significantly byTACKI. Meanwhile, the content of TNF-α and IL-6 in rat brain tissue is reduced. ConclusionTACKI has a significant analgesic activities. It may be related to inhibiting the activation of astrocytes and reducing the content of inflammatory mediators.

In-vivo evaluation of molybdenum as bioabsorbable stent candidate

Biodegradable stents have tremendous theoretical potential as an alternative to bare metal stents and drug-eluting stents for the treatment of obstructive coronary artery disease. Any bioresorbable or biodegradable scaffold material needs to possess optimal mechanical properties and uniform degradation behavior that avoids local and systemic toxicity. Recently, molybdenum (Mo) has been investigated as a potential novel biodegradable material for this purpose. With its proven moderate degradation rate and excellent mechanical properties, Mo may represent an ideal source material for clinical cardiac and vascular applications. The present study was performed to evaluate the mechanical performance of metallic Mo in vitro and the biodegradation properties in vivo. The results demonstrated favorable mechanical behavior and a uniform degradation profile as desired for a new generation ultra-thin degradable endovascular stent material. Moreover, Mo implants in mouse arteries avoided the typical cellular response that contributes to restenosis. There was minimal neointimal hyperplasia over 6 months, an absence of excessive smooth muscle cell (SMC) proliferation or inflammation near the implant, and avoidance of significant harm to regenerating endothelial cells (EC). Qualitative inspection of kidney sections showed a potentially pathological remodeling of kidney Bowman's capsule and glomeruli, indicative of impaired filtering function and development of kidney disease, although quantifications of these morphological changes were not statistically significant. Together, the results suggest that the products of Mo corrosion may exert beneficial or inert effects on the activities of inflammatory and arterial cells, while exerting potentially toxic effects in the kidneys that warrant further investigation.

Abstract 9749: High-Avidity Binding Drives Nucleation of Amyloidogenic Transthyretin Monomer in Cardiac Amyloidosis

Amyloidosis involves stepwise growth of fibrils assembled from soluble precursor proteins. Mutations of plasma carrier protein transthyretin (TTR) are associated with hereditary transthyretin amyloidosis (ATTR). TTR naturally folds into a stable tetramer for its function, whereas conditions and mutations that foster aberrant monomer formation facilitate TTR oligomeric aggregation and subsequent fibril extension. We investigated the dynamics of early assembly of oligomers by wild-type TTR, in comparison to its TTR V30M and TTR V122I disease mutants. We selected buffer conditions for monitoring time-dependent redistribution among monomer, dimer, tetramer, and oligomer contents in the presence and absence of multimeric TTR seeds. The seeds were artificially constructed recombinant multimers that contained 20-40 TTR subunits via engineered biotin-streptavidin interactions. As expected, these multimer seeds rapidly nucleated TTR monomers and dimers into larger complexes, while having no effect on tetramers. In vivo , streptavidin-induced multimers formed ATTR-like deposits in the heart and the kidney following i.v. injection in mice. While all three variants prominently deposited in the kidney glomerulus, only TTR V30M resulted in extensive deposition in the heart. The cardiac TTR deposits varied in size and shape and were localized to the intermyofibrillar space along the capillaries. These results are consistent with the notion of monomeric TTR to engage high-avidity interactions with tissue amyloids. Our multimeric induction approach therefore provides a new model for studying the initiation of TTR deposition in the heart.

Post-translational modification of amyloid a protein in patients with AA amyloidosis

AA amyloidosis is a disease caused by extracellular deposition of insoluble β-pleated sheet fibrils composed of amyloid A (AA) protein, an amino (N)-terminal fragment of serum amyloid A (SAA). The deposits disrupt tissue structure and compromise organ function. Although the disease is systemic, deposition in kidney glomeruli is the most common manifestation. The leading cause of AA amyloidosis is sustained or recurrent inflammation accompanied by elevated levels of SAA. Factors determining the conversion of SAA to AA amyloid fibrils have yet to be fully resolved. Herein, we present liquid chromatography tandem-mass spectrometry (LC-MS/MS) analysis of AA proteins purified from eight patients with AA amyloidosis. For the first time, post-translational modifications (PTM), including carbamylation, acetylation and oxidation, were identified on AA peptides; all eight samples showed some degree of PTM. The amyloid in 6 samples comprised peptides derived from SAA1 with few or none from SAA2, while the other two samples contained both SAA1- and SAA2-derived peptides. N-terminal AA peptides beginning with Arg1 as well as AA peptides starting with Ser2 were present in five of the eight samples, while all or nearly all of the N-terminal peptides in the other three samples lacked Arg1. These data demonstrate that multiple species of AA amyloid proteins can comprise the subunits in amyloid fibrils and raise the possibility that PTM may play a role in fibrillogenesis.

Antibiotic Dosing in Chronic Kidney Disease

Infectious process is an important cause of morbidity and mortality among patients with chronic kidney disease. Prescription of antibacterial drugs should take into account the pharmacokinetic parameters of the medicine and the individual characteristics of the patient. Adequate antibiotic dosing is crucial for positive treatment outcome and minimisation of side effects. The aim of the study was to analyse scientific literature on factors affecting the dosing of antibacterials in patients with chronic kidney disease. Since most antibacterial medicines are eliminated by the kidneys, a decrease in glomerular filtration rate or kidney function should be followed by the dose adjustment in order to prevent the medicine accumulation and reduce the risk of side effects. Antibiotic dosing in such patients should be accompanied by kidney function assessment and be adjusted to ensure effective and safe treatment, as well as prevention of bacterial resistance. The review provides data on the dosing of some antibiotic groups (beta-lactams, aminoglycosides, fluoroquinolones) at different creatinine clearance rates. Extrarenal excretion of medicines does not usually require the dose adjustment in patients with chronic kidney disease.

Imaging intact human organs with local resolution of cellular structures using hierarchical phase-contrast tomography

Imaging intact human organs from the organ to the cellular scale in three dimensions is a goal of biomedical imaging. To meet this challenge, we developed hierarchical phase-contrast tomography (HiP-CT), an X-ray phase propagation technique using the European Synchrotron Radiation Facility (ESRF)’s Extremely Brilliant Source (EBS). The spatial coherence of the ESRF-EBS combined with our beamline equipment, sample preparation and scanning developments enabled us to perform non-destructive, three-dimensional (3D) scans with hierarchically increasing resolution at any location in whole human organs. We applied HiP-CT to image five intact human organ types: brain, lung, heart, kidney and spleen. HiP-CT provided a structural overview of each whole organ followed by multiple higher-resolution volumes of interest, capturing organotypic functional units and certain individual specialized cells within intact human organs. We demonstrate the potential applications of HiP-CT through quantification and morphometry of glomeruli in an intact human kidney and identification of regional changes in the tissue architecture in a lung from a deceased donor with coronavirus disease 2019 (COVID-19).

Dynamics of urinary and respiratory shedding of Severe acute respiratory syndrome virus 2 (SARS-CoV-2) RNA excludes urine as a relevant source of viral transmission

PurposeTo investigate the expression of the receptor protein ACE-2 alongside the urinary tract, urinary shedding and urinary stability of SARS-CoV-2 RNA.MethodsImmunohistochemical staining was performed on tissue from urological surgery of 10 patients. Further, patients treated for coronavirus disease (COVID-19) at specialized care-units of a university hospital were assessed for detection of SARS-CoV-2 RNA in urinary samples via PCR, disease severity (WHO score), inflammatory response of patients. Finally, the stability of SARS-CoV-2 RNA in urine was analyzed.ResultsHigh ACE-2 expression (3/3) was observed in the tubules of the kidney and prostate glands, moderate expression in urothelial cells of the bladder (0–2/3) and no expression in kidney glomeruli, muscularis of the bladder and stroma of the prostate (0/3). SARS-CoV-2 RNA was detected in 5/199 urine samples from 64 patients. Viral RNA was detected in the first urinary sample of sequential samples. Viral RNA load from other specimen as nasopharyngeal swabs (NPS) or endotracheal aspirates revealed higher levels than from urine. Detection of SARS-CoV-2 RNA in urine was not associated with impaired WHO score (median 5, range 3–8 vs median 4, range 1–8, p = 0.314), peak white blood cell count (median 24.1 × 1000/ml, range 5.19–48.1 versus median 11.9 × 1000/ml, range 2.9–60.3, p = 0.307), peak CRP (median 20.7 mg/dl, 4.2–40.2 versus median 11.9 mg/dl, range 0.1–51.9, p = 0.316) or peak IL-6 levels (median: 1442 ng/ml, range 26.7–3918 versus median 140 ng/ml, range 3.0–11,041, p = 0.099). SARS-CoV-2 RNA was stable under different storage conditions and after freeze–thaw cycles.ConclusionsSARS-CoV-2 RNA in the urine of COVID-19 patients occurs infrequently. The viral RNA load and dynamics of SARS-CoV-2 RNA shedding suggest no relevant route of transmission through the urinary tract.

GYY4137 Regulates Extracellular Matrix Turnover in the Diabetic Kidney by Modulating Retinoid X Receptor Signaling.

Diabetic kidney is associated with an accumulation of extracellular matrix (ECM) leading to renal fibrosis. Dysregulation of retinoic acid metabolism involving retinoic acid receptors (RARs) and retinoid X receptors (RXRs) has been shown to play a crucial role in diabetic nephropathy (DN). Furthermore, RARs and peroxisome proliferator-activated receptor γ (PPARγ) are known to control the RXR-mediated transcriptional regulation of several target genes involved in DN. Recently, RAR and RXR have been shown to upregulate plasminogen activator inhibitor-1 (PAI-1), a major player involved in ECM accumulation and renal fibrosis during DN. Interestingly, hydrogen sulfide (H2S) has been shown to ameliorate adverse renal remodeling in DN. We investigated the role of RXR signaling in the ECM turnover in diabetic kidney, and whether H2S can mitigate ECM accumulation by modulating PPAR/RAR-mediated RXR signaling. We used wild-type (C57BL/6J), diabetic (C57BL/6-Ins2Akita/J) mice and mouse mesangial cells (MCs) as experimental models. GYY4137 was used as a H2S donor. Results showed that in diabetic kidney, the expression of PPARγ was decreased, whereas upregulations of RXRα, RXRβ, and RARγ1 expression were observed. The changes were associated with elevated PAI-1, MMP-9 and MMP-13. In addition, the expressions of collagen IV, fibronectin and laminin were increased, whereas elastin expression was decreased in the diabetic kidney. Excessive collagen deposition was observed predominantly in the peri-glomerular and glomerular regions of the diabetic kidney. Immunohistochemical localization revealed elevated expression of fibronectin and laminin in the glomeruli of the diabetic kidney. GYY4137 reversed the pathological changes. Similar results were observed in in vitro experiments. In conclusion, our data suggest that RXR signaling plays a significant role in ECM turnover, and GYY4137 modulates PPAR/RAR-mediated RXR signaling to ameliorate PAI-1-dependent adverse ECM turnover in DN.

Contactin-1 is a novel target antigen in membranous nephropathy associated with chronic inflammatory demyelinating polyneuropathy

Primary membranous nephropathy (MN) is an autoimmune glomerular disease in which autoantibodies are directed against podocyte proteins. In about 80% of cases the main targeted antigen is the phospholipase A2 receptor 1 (PLA2R1). Anti-PLA2R1 antibodies are mainly immunoglobulin G type 4 (IgG4). However, the antigenic target remains to be defined in 20% of cases. MN can be associated with chronic inflammatory demyelinating polyneuropathy, an autoimmune disease of the peripheral nervous system where a common antigenic target has yet to be identified. To ascertain a possible novel target antigen, we analyzed kidney biopsies from five patients positive for anti-contactin 1 antibodies and presenting with MN combined with chronic inflammatory demyelinating polyneuropathy. Eluted IgG from biopsy sections against contactin 1 and nerve tissue were screened. Western blot revealed contactin 1 expression in normal kidney glomeruli. Confocal microscopic analysis showed the presence and colocalization of contactin 1 and IgG4 on the glomerular basement membrane of these patients. Glomerular contactin 1 was absent in patients with anti-PLA2R1-associated MN or membranous lupus nephritis or a healthy control. The eluted IgG from contactin 1-positive biopsy sections but not the IgG eluted from patients with PLA2R1 MN bound contactin 1 with the main eluted subclass IgG4. Eluted IgG could bind paranodal tissue (myelinated axon) and colocalized with commercial anti-contactin 1 antibody. Thus, contactin 1 is a novel common antigenic target in MN associated with chronic inflammatory demyelinating polyneuropathy. However, the precise pathophysiology remains to be elucidated.

Aqueous fruit pulp extract of Adansonia digitata (L) protects against lead-acetate-induced hepato-renal damage in rat model

BackgroundAdansonia digitata (L) fruit has a multi-purpose function one among many, is the antioxidant activities of the fruit by preventing oxidative stress. The effect of Adansonia digitata (L) fruit on lead-induced liver and kidney damage is not clear. Hence, the study was aimed to assessed the protective role of Adansonia digitata (L) fruits against lead acetate induced changes in the liver and kidney function test parameters and the histology of both organ in experimental rats. The rats were divided into five groups with five rats each. All the rats were administered with respective assigned treatment once daily for 6 weeks. Rats in groups I were administered with just distil water (2 ml/kg). Rats in groups II were administered with lead acetate (30 mg/kg) while rats in groups III–V were administered Adansonia digitatata (L) fruit extract (250 mg/kg and 500 mg/kg) and Succimer (5 mg/kg) respectively, then additionally challenged with lead acetate (30 mg/kg) immediately after. At the end of the administration, the blood serum from the experimental rats were used for biochemical analysis. Then, the the organs such as the liver and kidney collected for histological study.ResultsRats administered with Lead acetate showed an increase in AST, ALP and ALT as well as increase in urea and creatinine level (p < 0.001), when compared with the control group (group I), where as Adansonia digitatata (L) fruit prevented the effect (upsurge of serum, Urea, Creatinine, AST, ALP and ALT) of lead acetate. Rats administer with only Lead acetate revealed marked liver steatosis and the degeneration of the kidney glomerulus. The Adansonia digitatata (L) fruit extract and Succimer prevented the histological liver steatosis, as well as the degeneration of the glomerulus of the kidney cytoarchitecture.ConclusionThe findings in this study suggest that Adansonia digitata fruits extract has a protective potentials against lead acetate induced liver and kidney toxicity by preventing the upsurge of liver function enzymes and kidney function parameters. Hence, Adansonia digitata fruits can serve as a natural plant agent that can prevent hepato-renal toxicity. Therefore, Adansonia digitata holds future prospects in preclinical framework to ameliorate organs toxicity for oral therapeutic applications.

Особливості порушень функціонування нирок у дітей із системним червоним вовчаком та ювенільним ідіопатичним артритом

Актуальність. При системних захворюваннях сполучної тканини автоімунної природи ураження нирок в одних випадках є синдромом захворювання, а в інших — наслідком основного процесу. Мета роботи — встановити частоту та характер порушень функціонування нирок у дітей та підлітків, хворих на ювенільний ідіопатичний артрит (ЮІА) та системний червоний вовчак (СЧВ), залежно від тривалості та активності цих захворювань. Матеріали та методи. Обстежені 113 дітей у віці від 7 до 18 років, із них хворих на ЮІА — 70, 43 дитини — з СЧВ. Розподіл на групи проведений залежно від тривалості та активності захворювання. Стан функції нирок оцінювали за допомогою геморенальних проб (рівень креатиніну сироватки крові), визначення швидкості клубочкової фільтрації. Установлювали рівень протеїнурії в добовій сечі, а також вивчали концентраційну функцію нирок за допомогою аналізу сечі за Зимницьким. Статистична обробка проводилася за допомогою пакета статистичних програм Statgrafics 16.0. Результати. Так, у дітей з ЮІА при тривалості хвороби більше ніж 3 роки відбувалися зміни функції нирок, що виражалися протеїнурією, порушенням швидкості клубочкової фільтрації та зниженням концентраційної функції нирок. При вивченні частоти порушень залежно від активності патологічного процесу встановлено що у групі з 0 ст. було зниження як швидкості клубочкової фільтрації (25,0 %), так і концентраційної функції нирок (12,5 %). У групі з I ст. активності виявлені мікропротеїнурія (4,4 %), порушення швидкості клубочкової фільтрації (12,3 %) та зниження концентраційної функції нирок (6,6 %). У групі з II ст. також виявлені мікропротеїнурія (14,2 %) та певне зниження швидкості клубочкової фільтрації (14,2 %), а в групі з III ст. майже у всіх хворих зареєстровані мікропротеїнурія (66,6 %) та зниження концентраційної функції нирок (33,3 %). При вивченні функції нирок залежно від активності патологічного процесу привертали увагу приріст рівня мікропротеїнурії (р &lt; 0,01) та зниження клубочкової фільтрації в групі з III ступенем активності (р &lt; 0,01). Також установлено зростання рівня протеїнурії в групі з тривалістю захворювання понад 3 роки. Таким чином, установлено, що ураження нирок супроводжується значним зниженням клубочкової фільтрації та протеїнурією, тобто порушенням функції клубочкового апарату нирок. У хворих на СЧВ дітей із тривалістю захворювання від 1 до 3 років в 68,1 % випадків зареєстровані наявність протеїнурії та в поодиноких випадках — зміни швидкості клубочкової фільтрації. У групі дітей із тривалістю патологічного процесу понад 3 роки виявлено більш глибокі зміни функції нирок. Так, майже в усіх хворих установлена протеїнурія (90,4 %), частота зниження швидкості клубочкової фільтрації збільшилась до 19,0 %, у 14,2 % випадків встановлено зниження концентраційної функції нирок. При аналізі частоти порушень функції нирок залежно від активності СЧВ привертали увагу зростання рівня протеїнурії (р &lt; 0,05) та зниження швидкості клубочкової фільтрації (р &lt; 0,02) саме у хворих із II ступенем активності захворювання. При вивченні показників функції нирок залежно від тривалості захворювання на СЧВ установлено, що в групі з тривалістю хвороби понад 3 роки відбувається зниження швидкості клубочкової фільтрації (р &lt; 0,03), зростання рівня мікроальбумінурії (р &lt; 0,03) та зниження максимальної щільності сечі (р &lt; 0,03). При дослідженні показників функції нирок залежно від активності патологічного процесу виявлено зростання рівня мікроальбумінурії при II ступені активності (р &lt; 0,01). Таким чином, майже у третини дітей із СЧВ із тривалістю захворювання понад 3 роки та збереженням активності патологічного процесу відбувається формування ознак ураження нирок із порушенням функції як клубочкового, так і канальцевого апарату нирок, що надалі буде сприяти розвитку хронічної ниркової недостатності. Висновки. У дітей, хворих на ЮІА та на СЧВ, із тривалістю захворювання понад 3 роки при збереженні активності патологічного процесу формуються порушення функції нирок. Ознаками ураження нирок є наявність протеїнурії, зниження клубочкової фільтрації, зниження концентраційної здатності нирок.

Discovery of re-purposed drugs that slow SARS-CoV-2 replication in human cells.

COVID-19 vaccines based on the Spike protein of SARS-CoV-2 have been developed that appear to be largely successful in stopping infection. However, therapeutics that can help manage the disease are still required until immunity has been achieved globally. The identification of repurposed drugs that stop SARS-CoV-2 replication could have enormous utility in stemming the disease. Here, using a nano-luciferase tagged version of the virus (SARS-CoV-2-ΔOrf7a-NLuc) to quantitate viral load, we evaluated a range of human cell types for their ability to be infected and support replication of the virus, and performed a screen of 1971 FDA-approved drugs. Hepatocytes, kidney glomerulus, and proximal tubule cells were particularly effective in supporting SARS-CoV-2 replication, which is in-line with reported proteinuria and liver damage in patients with COVID-19. Using the nano-luciferase as a measure of virus replication we identified 35 drugs that reduced replication in Vero cells and human hepatocytes when treated prior to SARS-CoV-2 infection and found amodiaquine, atovaquone, bedaquiline, ebastine, LY2835219, manidipine, panobinostat, and vitamin D3 to be effective in slowing SARS-CoV-2 replication in human cells when used to treat infected cells. In conclusion, our study has identified strong candidates for drug repurposing, which could prove powerful additions to the treatment of COVID.

STUDY OF ACUTE AND SUB-ACUTE TOXICITY OF Boehmeria virgata (FORST) GUILL LEAF EXTRACT IN WISTAR RATS

This study was carried out to evaluate the acute and sub-acute toxicity of the standardized Boehmeria virgata leaf extract (BLVE) in Wistar rats. B. virgata is a traditional herb utilized by the people of Makassar, Indonesia to cure inflammation and cancer. In the current study, thirty (30) Wistar rats were divided into 6 groups (3 groups of males and 3 groups of females) for acute treatments. Similarly, for investigating sub-acute treatment forty (40) Wistar rats were split into 8 groups (4 groups of males and 4 groups of females). For acute toxicity treatment, selected rats received 2000 mg and 5000 mg/kg of BVLE by oral administration for 14 days while for the estimation of sub-acute toxicity, experimental rats were treated with 250, 500, and 1000 mg/kg BVLE for 28 days. After 14 days of treatment, the rats were monitored for any form of changes in behavior, weight, food, water intake, and histopathology. The treated animals underwent hematological, biochemical, histopathological, and organ weight analysis after 28 days. Results of the study revealed no significant differences in body and organ weight, intake of food, and water after acute BVLE treatment in rats compared with those in the control group. The histopathological study suggested a general hydropic degeneration of the liver after acute treatment with 5000 mg/kg; such degeneration did not occur in the kidneys and kidney glomerulus of BVLE treated rats. Further, no significant toxicity was shown in hematological, biochemical, organ weight, and histopathological data in the sub-acute BVLE group after comparing treated rats with the control group. Results of the study can be concluded that BVLE is not toxic at doses of up to 1000 mg/kg. Therefore B. virgate fulfilled a preclinical criterion that is necessary for its further establishment as a clinically useful extract.

Ендотеліальна дисфункція у патогенезі діабетичної хвороби нирок

У статті надана комплексна характеристика ролі гіперглікемії в патогенезі судинних ускладнень цукрового діабету. Проаналізовано сучасні дані щодо механізмів розвитку і прогресування діабетичної хвороби нирок. Охарактеризовано особливості будови і пошкодження ендотелію і базальної мембрани в капілярах клубочків нирок і оцінена роль ендотеліальної дисфункції в патогенезі діабетичної хвороби нирок. Пошук літератури проводився по базах даних Scopus, PubMed, MedLine і CyberLeninka.

Molecular identification and pathogenicity study of virulent Citrobacter freundii associated with mortality of farmed Labeo rohita (Hamilton 1822), in India

The present research study was carried out to investigate the causative agent responsible for infection and mortality of farmed Labeo rohita. Samples of infected fishes were collected from aquaculture farms of West Bengal, India. Biochemical and molecular identification revealed that isolated strain EM4 was Citrobacter freundii. Furthermore, a challenge study was carried out to determine the number of bacterial colonies required for 50% mortality (LD50) of the experimental fish population. LD50 of the pathogenic bacterial isolates (C. freundii) was found to be 1.2 × 103 CFU per fish. Histopathological studies revealed damaged glomerulus in kidney and necrosis of hepatocytes in liver of diseased L. rohita. Additionally, non-specific and specific immune responses of L. rohita were also studied after C. freundii challenge. At early stages of infection, elevated expression level of few non-specific responses viz. respiratory burst activity, myeloperoxidase activity and alpha 2 macroglobulin were observed. However, the antiprotease activity was found to be lowered at the early stages of infections. Sequential changes in the immune-related genes viz. complement component 3 (C3) and interleukin 6 (IL- 6) were also studied in the liver, kidney and muscle tissues of the artificially infected fishes. In the liver and muscle tissues, the C3 gene was significantly up-regulated at the early stages of infections, whereas the IL-6 expression in liver was found to be down-regulated throughout the infection process. Expression of C3 and IL-6 genes in kidney were found to be down-regulated at the early stages of infections, but later, it was up-regulated with the progression of the infection. The result of the present study suggests that C. freundii is a concerning threat towards developing aquaculture sector.The pathology and pathogenesis study will help to understand the infection process as well as in the development of therapies to control the infection.