Urine Biochemical Parameters Research Articles

Chronic Carriage of Leptospira interrogans Genotype Associated With the Australis Serogroup by Naturally-Infected Hedgehogs (Erinaceus europaeus) at a Wildlife Health Centre in Northwestern France.

Leptospirosis is a widespread zoonosis caused by bacteria in the genus Leptospira. Basic epidemiological information is crucial to mitigating disease risk but is lacking for leptospirosis; notably, the hosts responsible for maintaining Leptospira remain largely unknown. Frequently observed near human habitations, hedgehogs (Erinaceus europaeus) are taken to wildlife rescue centres when found sick or injured. Thus, they may pose a risk to human and animal health if they carry pathogenic Leptospira. This study aimed to describe Leptospira carriage in a hedgehog population and the potential clinical impacts of the infection. We investigated Leptospira carriage frequency and diversity in urine samples from 69 hedgehogs at a wildlife rescue centre, between April and June 2022. We used quantitative PCR, typing of the 16S rRNA and lfb1 genes, variable number tandem repeat and multispacer sequence typing to characterise Leptospira DNA. An analysis of urinary biochemical parameters was conducted to assess renal function. We detected Leptospira DNA in 25 (35%) of the urine samples, of which 21 were successfully typed. The latter analysis revealed a limited degree of genetic diversity. L. interrogans (n = 19) predominated, and the only genotype detected was related to the Australis serogroup (n = 17). We also noted the presence of L. borgpetersenii (n = 1) and L. kirschneri (n = 1). There was no relationship between infection status and urinalysis parameters. These results suggest hedgehogs may act as long-term shedders of Leptospira in natural ecosystems.

Effects of Exercise on Urinary Parameters and Proteins in Well-Trained Canicross Dogs: A Preliminary Study.

The aim of the present study was to investigate the effects of exercise during canicross competition on urinary biochemical parameters and urinary proteins. Our results showed that, among the parameters analysed in routine urinalysis, pH was affected by exercise and albuminuria also showed changes immediately after physical exercise with a return to baseline within 2 h after the physical activity. In particular, SDS-PAGE electrophoresis allowed the separation of the urinary proteins on the basis of their molecular mass allowing a qualitative evaluation of the urinary proteome and a quantitative analysis of urine albumin and uromodulin, with a significant increase in urinary albumin and a decrease in uromodulin after exercise. In this study, urine was collected by spontaneous voiding and the use of this non-invasive sampling method proved useful in obtaining information on the health of the animals in the immediate moments before and after the competition without increasing the stress of the animals involved in the competition by any visits or invasive manipulations by veterinarians.

Nephroprotective effect of Cansjera rheedii on Gentamicin induced Renal toxicity in Wistar albino rats

Purpose: To examine the effect of Cansjera rheedii in gentamicin induced kidney damage in rats. Material and Method: In this current study, rats in group 1 were used as the normal group and were given physiological saline. Rats of group 2 were injected with gentamicin [100mg/kg] via intraperitoneal injection. Group 3 was given gentamicin (100mg/kg, IP) together with oral selenium [2mg/kg]. Hydroalcoholic extract (HECR) of Cansjera rheedii leaves was administered orally to groups 4 and 5 in low dose of 200mg/kg and high dose of 400 mg/kg, respectively. Above treatments were given to rats over eight days. Blood serum and urine were collected on the 8th day to evaluate blood and urine biochemical parameters. Antioxidant parameters were evaluated using renal homogenates. Kidney sections were used for histopathological examination. Result: The renoprotective effect of HECR plant extract was evident at both low and high dose [200mg/kg and 400mg/kg]. This action maintains structural and functional integrity by preventing damage caused by gentamicin. This protection is attributed to the antioxidant properties of the extract.

Effect of aluminum hydroxide on serum phosphate and fibroblast growth factor 23 concentrations in young adult cats with surgically induced chronic kidney disease.

To describe serum fibroblast growth factor 23 (FGF-23) concentrations in young adult cats with remnant kidney model-induced chronic kidney disease (CKD) and to evaluate the effects of orally administered aluminum hydroxide (ALOH) on serum phosphate and FGF-23 concentrations in these cats. 17 adult, purpose-bred cats with induced CKD and 13 healthy, age-matched cats. A prospective, randomized study. Cats with induced CKD fed a wet renal diet received treatment with ALOH (90 mg/kg/d, PO) on days 0 to 42 and no treatment on days 43 to 84 (treatment group, n = 9) or no treatment on days 0 to 84 (control group, n = 8). Standard serum and urine biochemical analyses and several parameters reflective of calcium-phosphate balance, including serum parathyroid hormone and FGF-23 concentrations, were evaluated at baseline and various time points, including days 42 and 84. Age-matched, healthy, community-owned cats underwent similar evaluations at a single time point. Baseline data from CKD cats were compared to those of healthy cats. Longitudinal data from CKD cats were compared over time. Serum phosphate, total and ionized calcium, and FGF-23 concentrations were significantly higher in CKD cats at baseline relative to healthy cats (all P ≤ .009). Serum phosphate concentration did not change significantly over time in either CKD group; however, FGF-23 concentrations significantly increased over time in the control group (P < .02) but not the treatment group (P = .059). Aluminum hydroxide did not reduce serum phosphate or FGF-23 concentrations in this small study of cats with induced CKD chronically eating a phosphate-restricted diet.

Dapagliflozin restores autophagy and attenuates apoptosis via the AMPK/mTOR pathway in diabetic nephropathy rats and high glucose-induced HK-2 cells.

Diabetic nephropathy (DN) is a serious microvascular complication of diabetes mellitus. Significantly reduced levels of autophagy in diabetic kidneys play an important role in the development of DN. The present study investigated the effects of dapagliflozin (DAP) on renal autophagy and AMP-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) pathway in vivo and in vitro. We explored the effect of DAP in streptozotocin (STZ)-induced DN rats. The anti-DN effect of DAP was assessed by body weight, kidney weight/body weight ratio, blood and urine biochemical parameters, and pathological changes of kidney tissue. Number of autophagosomes in the kidney was investigated through Transmission electron microscopy. Besides, cell viability and apoptosis of DAP alone or combined with Compound C (CC, a selective AMPK inhibitor)-treated high glucose (HG)-induced HK-2 cells were detected by Cell Counting Kit-8 (CCK-8) and flow cytometry assays. Immunohistochemistry, Western blot, Enzyme-linked immunosorbent assay (ELISA), and immunofluorescence were employed to detect the expression levels of extracellular matrix (ECM) deposition, autophagy, apoptosis, and AMPK/mTOR pathway-associated targets in vivo and in vitro. The results showed that DAP ameliorated the body weight and decreased kidney weight, fasting blood glucose, and serum/urine biochemical parameters of renal damage, as well as renal pathological changes. Moreover, DAP significantly ameliorated HG-induced cell apoptosis and ECM deposition in HK-2 cells. However, these favorable effects of DAP could be abolished by co-treatment with CC in HG-induced HK-2 cells. Mechanistically, DAP can enhance autophagy in DN including increased LC3-II/I ratio, Beclin-1, p-AMPK protein levels, and decreased p62 and p-mTOR protein expressions, as well as inhibited renal fibrosis and apoptosis. In summary, DAP alleviated fibrosis, apoptosis, and autophagy in DN rats and HG-induced HK-2 cells by regulating the AMPK/mTOR pathway.

Evaluation of the thiazide challenge test to differentiate primary from hypercalciuria-related hyperparathyroidism.

treatment of primary hyperparathyroidism (PHPT) and secondary hyperparathyroidism due to idiopathic hypercalciuria (SHPT-IH) is markedly different. Robust diagnostic tools to differentiate between both entities are however lacking. evaluate the thiazide challenge test (TCT) in clinical practice, its aid in clinical decision making, evaluate the accuracy (sensitivity, specificity) and potentially useful parameters of the TCT. monocentric observational retrospective cohort study from January 2017 to November 2023. outpatient, Ghent University Hospital (Belgium). 25 adult patients with hypercalciuria, elevated parathyroid hormone (PTH), and high-normal or elevated serum calcium that underwent a TCT. TCT. serum, urinary biochemical parameters before and after testing, clinical and imaging outcomes, treatment, and follow-up. patients with a TCT-based working diagnosis of PHPT show greater increases in albumin-adjusted calcium and total serum calcium concentration than patients with SHPT-IH (+0,11 ± 0,10 vs. + 0,0071 ± 0,10mmol/l; p = 0,025 and +0,14 ± 0,12 vs. + 0,012 ± 0,15mmol/l; p = 0,024 respectively). The TCT-based working diagnosis of PHPT has a sensitivity of 81,8%, a specificity of 77,8% and a likelihood ratio of 3,68 of estimating a correct final diagnosis.Urinary calcium excretion, PTH, calcium-phosphorous ratio, PTH-inhibition rate, and parathyroid function index do not differ significantly in patients with PHPT compared to those with SHPT-IH. the TCT aids in discriminating patients with PHPT from those with SHPT-IH based on a rise in serum calcium. Other parameters are not different between both groups. Larger prospective trials are necessary to further define the diagnostic potential of the TCT, its most appropriate biochemical outcome variables, and decision cut-offs.

Preliminary study of urine analysis in pregnant cows: insights for improved herd management

ABSTRACT This study aimed to establish a standard urine profile for pregnant cows, commonly seen patients in veterinary clinics. Urine samples from 15 pregnant cows (6 Belgian Blue and 9 Holstein) of various ages were analysed, all showing no clinical signs of urinary tract disease. Samples were collected during preventive check-ups, either by spontaneous micturition or catheterisation. Macroscopic evaluation and urine dipstick tests revealed a mean pH of 8.17 ± 0.49. Moderate glycosuria and proteinuria were detected in two animals. No urobilinogen, ketone bodies, haemoglobin, or traces of blood were found. Laboratory urinalysis provided values (with 95% confidence intervals): osmolarity 493–1110.00 mOsm/l, sodium 52.00–122.00 mmol/l, calcium 1.28–3.98 mmol/l, potassium 76–144.00 mmol/l, magnesium 5.00–8.00 mmol/l, chlorine 40.00–80.00 mmol/l, protein 35.00–104.00 mg/l, glucose 0.40–1.27 mmol/l, creatinine 34.00–76.00 mmol/l, and urea 681.00–2086 mmol/l. These findings present the initial descriptive values of urine biochemical parameters in pregnant cows, emphasising their significance in assessing animal health. This research underscores its relevance by providing fundamental insights into animal health, offering a pathway to enhance agricultural sustainability and address food security, echoing the United Nations’ Sustainable Development Goals 1, 2, 8 and 12.

The COVID-19 Severity and Its Association with Intestinal Parasite Coinfection and Urine Biochemical Parameters among COVID-19-Confirmed Patients Admitted to Debre Markos University COVID-19 Center, Northwest Ethiopia.

Though most people with COVID-19 disease show asymptomatic to mild illness, a substantial number of patients are at high risk of developing severe disease and adverse outcomes with long COVID-19 and death. Even though some studies showed that previously existing infections with parasites amend the host's body defenses to increase resistance to infection with SARS-CoV-2, there is limited data in Ethiopia. This study is aimed at determining the COVID-19 disease severity and its association with intestinal parasite coinfection and urine biochemical parameters among COVID-19-confirmed patients admitted at Debre Markos University COVID-19 Center, 2021. A prospective cohort study was conducted on 136 RT-qPCR-confirmed COVID-19 patients admitted at Debre Markos University COVID-19 Center from January 1 to March 30, 2021. Sociodemographic and clinical data were collected by using standardized data collection forms. A urine biochemical test was performed using a dry urine dipstick kit and stool examination using direct wet mount microscopic examination and formalin-ether concentration method. The chi-square test, Fisher exact test, and ordinal logistic regression analysis were computed to assess association with outcome variables using Statistical Package for Social Science software (version 24). A total of 136 COVID-19-confirmed patients participated in this study. The median age of the participants was 48 years. The majority (86 (62.5%)) of them were male in sex. Of the 136 cases, 39 (28.7%) had died. Among the 136 patients, 22 (16.2%) were coinfected with intestinal parasites. COVID-19 patients who have intestinal parasite coinfection had lower odds of developing clinically severe COVID-19 compared to noninfected (AOR = 0.37; 95% CI = 0.147-0.944; P = 0.037). The majority (104 (76.5%)) of them have abnormal urine biochemical results. From the abnormal urine biochemical tests observed, the urine blood, glucose, and ketone tests were positive for 54 (39.7%), 36 (26.5%), and 30 (21.1%) patients, respectively. Among the 31 critical COVID-19 patients, 25 (80.6%) showed abnormal urine biochemical parameters. Age and comorbidity were significantly associated with COVID-19 severity (P < 0.05). Patients with old age and comorbidity had an increased risk of developing severe COVID-19 disease. Patients having SARS-CoV-2 and intestinal parasitic coinfections demonstrated mild COVID-19 disease severity. Abnormal urine biochemical results were common among critical COVID-19 patients. Thus, advanced study on the effect of the interaction among intestinal parasites on COVID-19 clinical severity and its mechanisms is essential.

Activation of TGR5 Increases Urine Concentration by Inducing AQP2 and AQP3 Expression in Renal Medullary Collecting Ducts

Introduction: G protein-coupled bile acid receptor (TGR5), the first G protein-coupled receptor for bile acids identified, is capable of activating a variety of intracellular signaling pathways after interacting with bile acids. TGR5 plays an important role in multiple physiological processes and is considered to be a potential target for the treatment of various metabolic diseases, including type 2 diabetes. Evidence has emerged that genetic deletion of TGR5 results in an increase in basal urine output, suggesting that it may play a critical role in renal water and salt reabsorption. The present study aims to elucidate the effect and mechanism of TGR5 activation on urine concentration. Methods: Mice were treated with TGR5 agonists (LCA and INT-777) for 3 days. The 24-h urine of mice was collected and analyzed for urine biochemical parameters. The mRNA expressions were detected by real-time PCR, and the protein expressions were detected by western blot. Immunohistochemistry and immunofluorescence were performed to examine the cellular location of proteins. The cultured primary medullary collecting duct cells were pretreated with H89 (a PKA inhibitor) for 1 h, followed by 12-h treatment of LCA and INT-777. Luciferase reporter assays were used to detect the effect of CREB on the gene transcription of AQPs. Gel electrophoretic mobility shift assays were used to analyze DNA–protein interactions. Results: Treatment of mice with the TGR5 agonist LCA and INT-777 markedly reduced urine output and increased urine osmolality, accompanied by a marked increase in AQP2 and AQP3 protein expression and membrane translocation. In cultured primary medullary collecting duct cells, LCA and INT-777 dose-dependently upregulated AQP2 and AQP3 expression in a cAMP/PKA-dependent manner. Mechanistically, both AQP2 and AQP3 gene promoter contains a putative CREB-binding site, which can be bound and activated by CREB as assessed by both gene promoter-driven luciferase and gel shift assays. Conclusion: Collectively, our findings demonstrate that activation of TGR5 can promote urine concentration by upregulation of AQP2 and AQP3 expression in renal collecting ducts. TGR5 may represent an attractive target for the treatment of patients with urine concentration defect.

Chronopharmacology of diuresis via metabolic profiling and key biomarker discovery of the traditional Chinese prescription Ji-Ming-San using tandem mass spectrometry in rat models

BackgroundJi-Ming-Shan (JMS) is a traditional prescription used for patients with rheumatism, tendons swelling, relief of foot pain, athlete's foot, diuresis, gout. Although many studies have investigated the active compounds in each herb, the functional mechanism behind its therapeutic effect remains unclear. Study designMetabolic cages for sample collection. The serum components obtained from the experimental animals were analyzed using LC-MS/MS. Furthermore, cross-analysis using the software MetaboAnalyst and Venn diagrams were used to investigate chronopharmacology of JMS in the animal models. PurposeThe aim of this study is to analyze the diuretic effects of JMS and to explore their chronopharmacology involved in organ regulation through four-quarter periods from serum samples of rat models. MethodsMetabolic cages were used for collecting the urine samples and PocketChem UA PU-4010, Fuji DRI-CHEM 800 were used to examine the urine biochemical parameters. The serum components were identified through ultra-performance liquid chromatography-quadrupole time-of-flight (UPLC-Q-TOF) with a new developed method. Cross analysis, Venn diagram, MetaboAnalyst were used to investigate the key biomarker and major metabolism route with the oral administration of the drug. ResultJMS significantly changed the 6 h urine volume with no observed kidney toxicity. Urine pH value ranges from 7.0 to 7.5. The chronopharmacology of JMS diuresis activity were 0–6 and 6–12 groups. UPLC-Q-TOF analyses identified 243 metabolites which were determined in positive mode and negative mode respectively. With cross analysis in the Venn diagram, one key biomarker naringenin-7-O-glucoside has been identified. Major metabolic pathways such as 1: Glycerophospholipid metabolism, 2: Primary bile acid biosynthesis, 3: Sphingolipid metabolism, 4: Riboflavin metabolism, 5: Linoleic acid metabolism, 6: Butanoate metabolism. ConclusionJMS significantly changed the urine output of animals in the 0–6 and 6–12 groups. No change in urine pH was observed and also kidney toxicity. A new UPLC-Q-TOF method was developed for the detection of the metabolites of JMS after oral administration. The cross analysis with Venn diagram and identified the key biomarker of JMS namely naringenin-7-O-glucoside. The results showed that six major pathways are involved in the gastrointestinal system and the liver. This study demonstrated the capability of JMS prescription in the regulation of diuresis and identified a key biomarker that is responsible for its therapeutic effect.

Improvement of microvascular complications in STZ-diabetic rats treated with Pterocarpus erinaceus Poir. extract

Pterocarpus erinaceus Poir. from Fabaceae family is a medicinal plant traditionally used in decoction or infusion to treat diabetes mellitus. Although this plant is used in treating diabetes, studies on the effectiveness of its stem bark on the complications induced by chronic hyperglycemia have not been thoroughly addressed. Thus, this study was conducted to prove the efficacy of hydroethanolic extract of stem bark of P. erinaceus on type 2 diabetes and its complications, such as renal fibrosis and retinopathy in rats. STZ diabetics. The dry extract of P. erinaceus stem bark was obtained following the hydroethanolic extraction (v/v). Diabetes was induced with streptozocin in SD rats pretreated with fructose-lard for 20 days. Then, the serum and urinary biochemical parameters were evaluated at the start and the end of the treatment. Rats with blood glucose ≥350 mg/dL and significant proteinuria were selected and treated with P. erinaceus stem bark extract and glibenclamide for 3 weeks. A complete blood count and a histopathological examination of the retina and kidneys were performed at the end of the 41st day of treatment. The results showed that P. erinaceus extract at a dose of 500 mg/kg bw and glibenclamide at a dose of 0.6 mg/kg bw caused a significant decrease (p < 0.0001) in basal blood glucose in STZ diabetic rats during treatment and improved oral glucose intolerance. At the end of the experiment, the treated rats showed a normalization in body weight, food and water consumption. Evaluating of biochemical parameters showed a significant (p < 0.001) decrease in total cholesterol, LDL-C, triglycerides, TG/HDL-C ratio, CPK and oxidative stress in treated rats. No retinal and kidney abnormalities were observed on histological sections in rats treated with plant extract and glibenclamide. In contrast, macular edema and renal fibrosis were observed in the diabetic control group. The findings showed that extract at a dose of 500 mg/kg bw improves oral glucose intolerance, and inhibits lipid deposition and retinal and renal fibrosis. Therefore, the plant extract could be exploited in the production of herbal medicines to manage diabetes and its complications.

Integrated multi-omics and bioinformatic methods to reveal the mechanisms of sinomenine against diabetic nephropathy

ObjectivesDiabetic Nephropathy (DN) is a serious complication of diabetes, the diagnosis and treatment of DN is still limited. Sinomenine (SIN) is an active extract of herbal medicine and has been applied into the therapy of DN.MethodsIn the part of bioinformatic analyses, network pharmacology and molecular docking analyses were conducted to predict the important pathway of SIN treatment for DN. In-vivo study, DN rats were randomized to be treated with vehicle or SIN (20 mg/kg or 40 mg/kg) daily by gavage for 8 weeks. Then, the pharmacological effect of SIN on DN and the potential mechanisms were also evaluated by 24 h albuminuria, histopathological examination, transcriptomics, and metabolomics.ResultsFirstly, network pharmacology and molecular docking were performed to show that SIN might improve DN via AGEs/RAGE, IL-17, JAK, TNF pathways. Urine biochemical parameters showed that SIN treatment could significantly reduce 24 h albuminuria of DN rats. Transcriptomics analysis found SIN could affect DN progression via inflammation and EMT pathways. Metabolic pathway analysis found SIN would mainly involve in arginine biosynthesis, linoleic acid metabolism, arachidonic acid metabolism, and glycerophospholipid metabolism to affect DN development.ConclusionsWe confirmed that SIN could inhibit the progression of DN via affecting multiple genes and metabolites related pathways.

Effects of exercise on urinary biochemical parameters and proteins in a group of well-trained military working dogs

Exercise-induced proteinuria has been widely investigated in humans, also in relation to intensity and duration of activity. Instead, there are only limited publications regarding urinary biochemical parameters and urinary proteins before and after physical activity in dogs. This paper aimed to investigate the effects of exercise on urinary biochemistry and proteins in military dogs. Twenty-four dogs were enrolled in this study. All the dogs were clinically sound, and they were examined before and after activity. Pulse rates (PR) and respiratory rate (RR) were monitored. Urine was sampled before and after a training session of search activity. Standard urinalysis was carried out, urine total proteins and creatinine were measured and the urinary protein:creatinine ratio was calculated; finally, the urinary proteins were separated using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Clinical examination before and after activity did not reveal any pathological finding. After activity, the PR was slightly increased, while the RR was notably increased (p < 0.05). Total proteins, albumin, and their ratio with creatinine were significantly higher after exercise when considering all the dogs included or only the females while, when considering only the males no significant difference was detected. The clinical relevance of this study was related to the possibility of using urine as a non-invasive sample for monitoring health status after training activity and exercise in dogs. An increase in microalbuminuria after search activity, measured using SDS-PAGE could be considered an early biomarker of renal function during training sessions.

Why is childhood urolithiasis increasing? Etiology, diagnosis and management: a single-center experience.

Globally, urolithiasis is becoming more and more common among children. We aimed to determine the etiology, and the diagnostic and therapeutic approaches in patients with urolithiasis. This was a retrospective study which included all patients (aged 1month-18years) admitted to the pediatric nephrology clinic in Elazığ Fethi Sekin City Hospital with urolithiasis between November 2019 and 2021. Only patients whose diagnosis of urolithiasis was confirmed by urinary ultrasonography were included in the study, while patients with chronic diseases (neurological diseases such as epilepsy, cerebral palsy, chronic bowel diseases, etc.) predisposing to kidney stone formation were not. Demographic characteristics, serum and urine biochemical parameters, urine metabolic and kidney stone metabolic and chemical analyses, urinary tract ultrasonography findings and treatment modalities were collected. One hundred ninety-seven patients (91 female and 106 male) were included in the study. Hypervitaminosis D was detected in 4 (2%) patients, suppressed parathyroid hormone in 12 (6%) and hypercalcemia in 27 (14%) patients. Metabolic screening showed hypercalciuria in 69 (35%) patients, hypocitraturia in 39 (20%), hyperoxaluria in 15 (8%) and cystinuria in 6 (3%) patients. Eighty three (42%) patients had a positive family history for kidney stones. One hundred eighteen (60%) patients received potassium citrate treatment, 71 (36%) were given hydration and diet recommendations without medical treatment, 6 (3%) received tiopronin treatment, and 2 (1%) patients were treated surgically. Our study suggests that Vitamin D supplementation at doses higher than 400IU/day may be a risk factor for kidney stonesin children. We observed that mothers tend not to give water to infants who are breastfed or formula-fed in the first year of life. K-citrate treatment can be a good option for prevention and dissolution of stones by alkalinization.

IgM and C3 Deposition in Primary Focal Segmental Glomerulosclerosis (FSGS): A Clinical and Histopathological Spectrum.

Focal segmental glomerulosclerosis (FSGS) is a common renal disorder, characterized by progressive segmental sclerosis of renal glomeruli and clinical symptoms corresponding toproteinuria. Classically, it is not considered to be an antibody-mediated disease, however, IgM and C3 deposition may be seen in a subset of cases of FSGS. The impact of this immune deposition on histopathological features in renal core biopsies, on the urinary biochemical parameters, and the clinical outcomes, has not been previously investigated in our population. The aim of this study is to analyze the aforementioned parameters in patients with primary FSGS having antibody deposition as compared to those who do not have any antibody deposition. Some 155 patients diagnosed with FSGS were retrospectively enrolled in our study. The renal biopsies were reviewed for histopathological features and immunofluorescence (IF) findings of IgM and C3 glomerular deposition. These histological features were then compared with the biochemical parameters as well as the clinical outcomes of patients. The patients were assigned to Groups 1 and 2 based on the IF findings. The IgM and/or C3 glomerular deposition had a low incidence in patients with primary FSGS in our study (28.3%). Patients having IgM and C3 co-deposition had a significantly longer time duration since the onset of their clinical symptoms; active disease duration (42 months vs 22 months, p=0.049). The mean pre-treatment serum creatinine of patients with IgM and C3 co-deposition was 6.00 mg/dL as compared to 3.29 mg/dL in patients with no immune deposition (p=0.037). The immune deposition was associated with higher rates of segmental and global glomerulosclerosis, but this finding along with other evaluated histological parameters did not show statistical significance. The number of patients having IgM and/or C3 deposition and with active steroid use/renal dialysis was similar to patients having no IgM and/or C3 deposition. The IgM and/or C3 deposition in FSGS has a low incidence within and is not associated with any significant differences in histological parameters on renal core biopsies of patients from the Pakistani population. IgM and/or C3 deposition is also associated with a significantly longer duration of active disease and these patients may present with higher pre-treatment serum creatinine. Other biochemical parameters and clinical outcomes appear comparable between the groups based on the available clinical data.

Baicalin Exerts a Protective Effect in Diabetic Nephropathy by Repressing Inflammation and Oxidative Stress Through the SphK1/S1P/NF-κB Signaling Pathway.

Inflammation and oxidative stress contribute to the development of diabetic nephropathy (DN). Baicalin (BA) shows renal protection against DN through its anti-inflammatory and anti-oxidant properties. However, the molecular mechanism by which BA exerts the therapeutic effects on DN remains to be investigated. The db/db mice and high glucose (HG)-induced HK-2 cells were used as the in vivo and in vitro model of DN, respectively. The effects of BA were assessed by detecting the related blood and urine biochemical parameters, kidney histopathology, inflammatory cytokine production, oxidative stress indicators, and apoptosis. Cell viability and apoptosis were detected by CCK-8 assay and TUNEL assay, respectively. Related protein levels were measured by an immunoblotting method. In db/db model mice, BA reduced serum glucose concentration, decreased blood lipid levels, ameliorated kidney functions, and decreased histopathological changes in kidney tissues. BA also alleviated oxidative stress and inflammation in db/db mice. In addition, BA blocked the activation of sphingosine kinases type 1/sphingosine 1-phosphate (SphK1/S1P)/NF-κB pathway in db/db mice. In HK-2 cells, BA hindered HG-induced apoptosis, oxidative stress and inflammation, while overexpression of SphK1 or S1P could reverse these effects. BA alleviated HG-induced apoptosis, oxidative stress and inflammation in HK-2 cells through the S1P/NF-κB pathway. Furthermore, BA blocked the NF-κB signaling by diminishing p65 nuclear translocation via the SphK1/S1P pathway. Our study strongly suggests that BA protects against DN via ameliorating inflammation, oxidative stress and apoptosis through the SphK1/S1P/NF-κB pathway. This study provides a novel insight into the therapeutic effects of BA in DN.

Cucumis callosus (Rottl.) Cogn. fruit extract ameliorates calcium oxalate urolithiasis in ethylene glycol induced hyperoxaluric Rat model

Cucumis callosus dry fruits are traditionally used as folk remedy to treat conditions like urethral irritations, urine stoppage or dribbling and other urinary ailments of man in north-west India. But no study is reported to validate this ethnic practice of using Cucumis fruit in urolithiasis. To evaluate anti-urolithiatic potential of Cucumis, hyperoxaluria was induced in rats by supplying 0.75% ethylene glycol (EG) + 1% ammonium chloride (AC) in drinking water for 14 days. Anti-urolithiatic activity of Cucumis callosus hydro-ethanolic extract (CCHEE) was assessed by measuring blood and urine biochemical parameters, oxidative stress indices, histopathology and osteopontin (OPN) expression. Administration of EG-AC to rats caused hyperoxaluria, crystalluria, azotaemia, oxidant/antioxidant imbalance (increase in lipid peroxidation (LPO), and decrease in glutathione (GSH) and catalase (CAT)), up-regulation of OPN and calcium oxalate (CaOx) crystal deposition in kidney. Treatment of afflicted rats with Cucumis fruits extract restored renal function to a great extent (CCHEE group), testified by improvement of stated parameters. Findings demonstrate curative efficacy of Cucumis fruit extract in EG induced urolithiasis of rats. The restoration of renal function was possibly by regulating renal stone formation via reducing urinary oxalate excretion, correcting oxidant/antioxidant imbalances, and reduced expression of OPN. Hence, results of this study validate the ethnic practice of using Cucumis fruit and conclude that fruit extracts have beneficial effects on CaOx urolithiasis and renal function.

The relationship between the stone's composition and the biochemical parameters of blood and urine in patients with urolithiasis

Urolithiasis is the most painful and frequent urological disease of the urinary tract. This study is the first in Morocco that aimed to determine the relationship between the urinary stone's composition and the biochemical analysis of blood and daily urine.A total of 154 urinary stones from lithiasis patients was collected between December 2009 and July 2015 at the Hassan II University Hospital in Fez-Morocco. The stones were analysed morphologically and chemically, blood and urine biochemical parameters were established and the relationship between these latter and the stones composition was determined.The stones analysis shows that more than 63% of stones are calcium oxalate, infection stones were more common in patients under 16 years of age. Twenty-two percent of patients had a higher creatinine level and 13% of them had hypercalciuria. However, uric acid stone (UA) patient's group were older than those of whewellite (Wh) group (p = 0.004). The blood creatinine level was significantly high in the UA and weddellite (Wd) groups (p = 0.032), and a significant difference between the two groups, Wh and Wd, for the Calciuria/Creatininuria ratio (p = 0.032). The Principal Component Analysis (PCA) showed a correlation between 24-hour uricuria, 24-hour urine, 24-hour diuresis, 24-hour creatinine, and natriuresis on the one hand, and with calciuria, 24-hour calciuria and BMI on the other hand.These results show the probable effect of different biochemical parameters of urine and blood in certain stones types formation, and presents a contribution to understand the urolithiasis risk factors in order to limit the recurrence of kidney stones and long-term complications

Urine stabilization and normalization strategies favor unbiased analysis of urinary EV content

Urine features an ideal source of non-invasive diagnostic markers. Some intrinsic and methodological issues still pose barriers to its full potential as liquid biopsy substrate. Unlike blood, urine concentration varies with nutrition, hydration and environmental factors. Urine is enriched with EVs from urinary-genital tract, while its conservation, purification and normalization can introduce bias in analysis of EV subsets in inter-and intra-individual comparisons. The present study evaluated the methods that decrease such biases such as appropriate and feasible urine storage, optimal single-step EV purification method for recovery of proteins and RNAs from small urine volumes and a normalization method for quantitative analysis of urine EV RNAs. Ultracentrifugation, chemical precipitation and immuno-affinity were used to isolate EVs from healthy donors’ urine that was stored frozen or at room temperature for up to 6 months. Multiple urine biochemical and EV parameters, including particle count and protein content, were compared across urine samples. To this purpose nanoparticle tracking analysis (NTA) and protein assessment by BCA, ELISA and WB assays were performed. These measurements were correlated with relative abundances of selected EV mRNAs and miRNAs assessed by RT-PCR and ranked for the ability to reflect and correct for EV content variations in longitudinal urine samples. All purification methods enabled recovery and downstream analysis of EVs from as few as 1 ml of urine. Our findings highlight long term stability of EV RNAs upon urine storage at RT as well as excellent correlation of EV content in urine with some routinely measured biochemical features, such as total urine protein and albumin, but not creatinine most conventionally used for urine normalization. Comparative evaluation of mRNA and miRNAs in EV isolates revealed specific RNAs, in particular RNY4 and small miRNA panel, levels of which well reflected the inter-sample EV variation and therefore useful as possible post-analytical normalizers of EV RNA content. We describe some realistic urine processing and normalization solutions for unbiased readout of EV biomarker studies and routine clinical sampling and diagnostics providing the input for design of larger validation studies employing urine EVs as biomarkers for particular conditions and diseases.

N-Ethyl-N-Nitrosourea (ENU) induced Osteoporosis in Rats by inducing mutation in Collagen Type 1 protein–A Novel Method Development

Osteoporosis is a life-threatening condition that primarily affects women. Mice, rats, dogs, and rabbits have all been used as research animals. Bone strength, bone micro architectural characteristics, and osteoporosis risk are all affected by genetic factors. In this sequence, Type I collagen is the primary bone protein encoded by the COLIA1 and COLIA2 genes. The goal of the study is to develop new model for osteoporosis in rats by using N-Ethyl-N- Nitrosourea (ENU) which particularly targets COL1A1. Further, we treated the osteoporotic rats with Collagen type protein, in order to cross check the disease induction. And Rats induced with ENU, exhibited mutation in COL1A1 gene increased in the levels of both serum and urine biochemical parameters like calcium, phosphorus, ALP and creatinine. PCR analysis reported the mutation in COL1A1 gene. The use of animal models of osteoporosis is required to better understand the complex nature of this condition and to create new preventive and treatment approaches.