Increase In Excretion Research Articles

PTEN, MMP2, and NF-κB and Regulating MicroRNA-181 Aggravate Insulin Resistance and Progression of Diabetic Nephropathy: A Case-Control Study

Diabetic nephropathy (DN) is characterized by an increase in urinary albumin excretion, diabetic glomerular lesions, and a decline in glomerular filtration rate (GFR). We assessed the expression of phosphatase and tensin homolog (PTEN), nuclear factor kappa-β (NF-κB), matrix metalloproteinase-2 (MMP2), and microRNA-181 in healthy controls (HC), individuals with type 2 diabetes mellitus (T2DM) without nephropathy, and those with DN. Our study investigated the association between these genes, insulin resistance (IR), and eGFR to gain insight into their roles in the pathogenesis and progression of DN. Anthropometric measurements and biochemical tests were conducted on HC (N = 36), T2DM (N = 38) patients, and DN (N = 35) patients. We used real-time polymerase chain reaction (RT-PCR) for whole blood gene expression analysis and performed bioinformatics analyses, including protein–protein interaction, gene ontology, and co-expression networks. We compared our expression data with other GEO-Microarray datasets. Our study highlights the role of IR in the progression of nephropathy in T2DM via the PTEN-Akt-mTOR signalling pathway. We also observed a decreasing trend in the expression of MMP2 and PTEN and an increasing trend in the expression of NF-κB and miR-181b-5p with the progression of nephropathy to the severe stage. The dysregulated expression of MMP2, PTEN, NF-κB, and miR-181b-5p in patients with T2DM contributes to the progression of T2DM to DN by aggravating IR, inflammation, accelerating basement membrane thickening, mesangial matrix expansion, and renal fibrosis.

The effect of orally administered nitrate on renal function and blood pressure in a randomized, placebo-controlled, crossover study in healthy subjects

BackgroundSeveral studies have shown inorganic nitrate/nitrite to reduce blood pressure in both healthy subjects and hypertensive patients. An effect presumably caused through bioconversion to nitric oxide. However, studies on inorganic nitrate/nitrite have shown inconsistent results on renal functions such as GFR and sodium excretion. The current study investigated whether orally administered nitrate would decrease blood pressure and increase GFR and urinary sodium excretion. MethodsIn a randomized, placebo-controlled, double-blinded, crossover study, 18 healthy subjects received a daily dose of 24 mmol potassium nitrate and placebo (potassium chloride) during 4 days in a randomized order. Subjects also ingested a standardized diet and completed a 24-h urine collection. GFR was determined by the constant infusion technique and during GFR measurement, brachial blood pressure (BP) and central blood pressure (cBP), heart rate, and arterial stiffness were measured every half hour using the Mobil-O-Graph®. Blood samples was analyzed for nitrate, nitrite, cGMP, vasoactive hormones and electrolytes. Urine was analyzed for nitrate, nitrite, cGMP, electrolytes, ENaCγ, NCC, CrCl, CH2O and UO. ResultsNo differences in GFR, blood pressure or sodium excretion were found between the treatments with potassium nitrate and placebo. However, both nitrate and nitrite levels in plasma and urine were significantly increased by potassium nitrate intake and the 24-h urinary excretion of sodium and potassium were stable, showing adherence to the standardized diet and the study medication. ConclusionWe found no decrease in blood pressure or increase in GFR and sodium excretion of 24 mmol potassium nitrate capsules as compared to placebo after 4 days of treatment.Healthy subjects may be able to compensate the effects of nitrate supplementation during steady state conditions. Future research should focus on long-term studies on the difference in response between healthy subjects and patients with cardiac or renal disease.

Role of Klotho and AGE/RAGE-Wnt/β-Catenin Signalling Pathway on the Development of Cardiac and Renal Fibrosis in Diabetes.

Fibrosis plays an important role in the pathogenesis of long-term diabetic complications and contributes to the development of cardiac and renal dysfunction. The aim of this experimental study, performed in a long-term rat model, which resembles type 1 diabetes mellitus, was to investigate the role of soluble Klotho (sKlotho), advanced glycation end products (AGEs)/receptor for AGEs (RAGE), fibrotic Wnt/β-catenin pathway, and pro-fibrotic pathways in kidney and heart. Diabetes was induced by streptozotocin. Glycaemia was maintained by insulin administration for 24 weeks. Serum and urine sKlotho, AGEs, soluble RAGE (sRAGE) and biochemical markers were studied. The levels of Klotho, RAGEs, ADAM10, markers of fibrosis (collagen deposition, fibronectin, TGF-β1, and Wnt/β-catenin pathway), hypertrophy of the kidney and/or heart were analysed. At the end of study, diabetic rats showed higher levels of urinary sKlotho, AGEs and sRAGE and lower serum sKlotho compared with controls without differences in the renal Klotho expression. A significant positive correlation was found between urinary sKlotho and AGEs and urinary albumin/creatinine ratio (uACR). Fibrosis and RAGE levels were significantly higher in the heart without differences in the kidney of diabetic rats compared to controls. The results also suggest the increase in sKlotho and sRAGE excretion may be due to polyuria in the diabetic rats.

Pathophysiology and clinical management of hyperkalemia in chronic kidney disease.

Adaptive increases in kidney and gastrointestinal excretion of K+ help to prevent hyperkalemia in patients with chronic kidney disease (CKD) as long as the glomerular filtration rate (GFR) remains >15-20 mL/min. K+ balance is maintained by increased secretion per functioning nephron, which is mediated by elevated plasma K+ concentration, aldosterone, increased flow rate, and enhanced Na+-K+-ATPase activity. Fecal losses of potassium also increase in CKD. These mechanisms are effective in preventing hyperkalemia if urine output is in excess of 600 mL/day and the GFR exceeds 15 mL/min. Development of hyperkalemia with only mild to moderate reductions in GFR should prompt a search for intrinsic disease of the collecting duct, disturbances in mineralocorticoid activity, and/or decreased delivery of sodium to the distal nephron. The initial approach to treatment is to review the patient's medication profile and whenever possible discontinue drugs that impair kidney K+ excretion. Patients should be educated on sources of K+ in the diet and should be strongly encouraged to avoid the use of K+ containing salt substitutes as well as herbal remedies since herbs may be a hidden source of dietary K+. Effective diuretic therapy and correction of metabolic acidosis are effective strategies to minimize the potential for hyperkalemia. Discontinuation or use of submaximal doses of renin-angiotensin blockers should be discouraged given the cardiovascular protective effect these drugs provide. Potassium binding drugs can be useful to enable use of these drugs and potentially allow liberalization of the diet in CKD patients.

Proteinuria Modifies the Relationship Between Urinary Sodium Excretion and Adverse Kidney Outcomes: Findings From KNOW-CKD

High sodium intake is associated with increased proteinuria. Herein, we investigated whether proteinuria could modify the association between urinary sodium excretion and adverse kidney outcomes in patients with chronic kidney disease (CKD). In this prospective observational cohort study, we included 967 participants with CKD stages G1 to G5 between 2011 and 2016, who measured 24-hour urinary sodium and protein excretion at baseline. The main predictors were urinary sodium and protein excretion levels. The primary outcome was CKD progression, which was defined as a≥50% decline in the estimated glomerular filtration rate (eGFR) or the onset of kidney replacement therapy. During a median follow-up period of 4.1 years, the primary outcome events occurred in 287 participants (29.7%). There was a significant interaction between proteinuria and sodium excretion for the primary outcome (P= 0.006). In patients with proteinuria of<0.5 g/d, sodium excretion was not associated with the primary outcome. However, in patients with proteinuria of≥0.5 g/d, a 1.0 g/d increase in sodium excretion was associated with a 29% higher risk of adverse kidney outcomes. Moreover, in patients with proteinuria of≥0.5 g/d, the hazard ratios (HRs) (95% confidence intervals[CIs]) for sodium excretion of<3.4 and≥3.4 g/d were 2.32 (1.50-3.58) and 5.71 (3.58-9.11), respectively, compared with HRs for patients with proteinuria of<0.5 g/d and sodium excretion of<3.4 g/d. In sensitivity analysis with 2 averaged values of sodium and protein excretion at baseline and third year, the results were similar. Higher urinary sodium excretion was more strongly associated with an increased risk of adverse kidney outcomes in patients with higher proteinuria levels.

Endothelial Progenitor Cells Affect the Growth and Apoptosis of Renal Cells by Secreting Microvesicles Carrying Dysregulated miR-205 and miR-206.

This study investigated the mechanism of microRNA (miRNA, miR) in microvesicles (MVs) secreted by endothelial progenitor cells (EPCs) involved in renal function in vivo and in vitro injury repair of rat primary kidney cells (PRKs). Gene Expression Omnibus analysis of potential target miRNAs in nephrotic rats. Real-time quantitative polymerase chain reaction verified the correlation of these miRNAs and screened the effective target miRNAs and their downstream putative target mRNAs. Western blot analyzes the protein levels of DEAD-box helicase 5 (DDX5) and the activation of the proapoptotic factor caspase-3/9 (cleaved). Dil-Ac-LDL staining, immunofluorescence, and a transmission electron microscope (TEM) were used to identify the successful isolation of EPCs and PRKs and the morphology of MVs. Cell Counting Kit-8 was used to detect the effect of miRNA-mRNA on the proliferation of PRKs. Standard biochemical kits were used to detect biochemical indicators in rat blood and urine. Dual-luciferase analysis of miRNA binding to mRNA was conducted. The effect of miRNA-mRNA interaction on the apoptosis level of PRKs was analyzed by flow cytometry. A total of 13 rat-derived miRNAs were potential therapeutic targets, and miR-205 and miR-206 were screened as the targets of this study. We found that the EPC-MVs alleviated the increase of blood urea nitrogen and urinary albumin excretion and the decrease in creatinine clearance caused by hypertensive nephropathy in vivo. The effect of MVs in improving renal function indicators was promoted by miR-205 and miR-206 and inhibited by knockdown of expressed miR-205 and miR-206. In vitro, angiotensin II (Ang II) promoted growth inhibition and apoptosis of PRKs, and similarly, dysregulated miR-205 and miR-206 affected the induction of Ang II. We then observed that miR-205 and miR-206 cotargeted the downstream target DDX5 and regulated its transcriptional activity and translational levels, while also reducing the activation of proapoptotic factors caspase-3/9. Overexpressed DDX5 reversed the effects of miR-205 and miR-206. By upregulating the expression of miR-205 and miR-206 in MVs secreted by EPC, the transcriptional activity of DDX5 and the activation of caspase-3/9 can be inhibited, thereby promoting the growth of PRKs and protecting the injury caused by hypertensive nephropathy.

Renoprotective effects of empagliflozin are linked to activation of the tubuloglomerular feedback mechanism and blunting of the complement system.

The mechanisms of nephroprotection in nondiabetic chronic kidney disease (CKD) models by sodium-glucose cotransporter 2 (SGLT2) inhibitors are not well defined. Five groups were established: sham-operated rats, placebo-treated rats with 5/6 nephrectomy (5/6Nx), 5/6Nx + telmisartan (5 mg/kg/day), 5/6Nx + empagliflozin (3 mg/kg/day), and 5/6Nx + empagliflozin (15 mg/kg/day). Treatment duration was 95 days. Empagliflozin showed a dose-dependent beneficial effect on the change from baseline of creatinine clearance (Ccr). The urinary albumin-to-creatinine ratio likewise improved in a dose-dependent manner. Both dosages of empagliflozin improved morphological kidney damage parameters such as renal interstitial fibrosis and glomerulosclerosis. 5/6 nephrectomy led to a substantial reduction of urinary adenosine excretion, a surrogate parameter of the tubuloglomerular feedback (TGF) mechanism. Empagliflozin caused a dose-dependent increase in urinary adenosine excretion. The urinary adenosine excretion was negatively correlated with renal interstitial fibrosis and positively correlated with Ccr. Immunofluorescence analysis revealed that empagliflozin had no effect on CD8+ and CD4+ T cells as well as on CD68+ cells (macrophages). To further explore potential mechanisms, a nonhypothesis-driven approach was used. RNA sequencing followed by quantitative real-time polymerase chain reaction revealed that complement component 1Q subcomponent A chain (C1QA) as well as complement component 1Q subcomponent C chain (C1QC) gene expression were upregulated in the placebo-treated 5/6Nx rats and this upregulation was blunted by treatment with empagliflozin. In conclusion, empagliflozin-mediated nephroprotection in nondiabetic CKD is due to a dose-dependent activation of the TGF as well as empagliflozin-mediated effects on the complement system.

Effects of elevated CO2 on metabolic rate and nitrogenous waste handling in the early life stages of yellowfin tuna (Thunnus albacares)

Ocean acidification is predicted to have a wide range of impacts on fish, but there has been little focus on broad-ranging pelagic fish species. Early life stages of fish are thought to be particularly susceptible to CO2 exposure, since acid-base regulatory faculties may not be fully developed. We obtained yellowfin tuna (Thunnus albacares) from a captive spawning broodstock population and exposed them to control or 1900 μatm CO2 through the first three days of development as embryos transitioned into yolk sac larvae. Metabolic rate, yolk sac depletion, and oil globule depletion were measured to assess overall energy usage. To determine if CO2 altered protein catabolism, tissue nitrogen content and nitrogenous waste excretion were quantified. CO2 exposure did not significantly impact embryonic metabolic rate, yolk sac depletion, or oil globule depletion, however, there was a significant decrease in metabolic rate at the latest measured yolk sac larval stage (36 h post fertilization). CO2-exposure led to a significant increase in nitrogenous waste excretion in larvae, but there were no differences in nitrogen tissue accumulation. Nitrogenous waste accumulated in embryos as they developed but decreased after hatch, coinciding with a large increase in nitrogenous waste excretion and increased metabolic rate in newly hatched larvae. Our results provide insight into how yellowfin tuna are impacted by increases in CO2 in early development, but more research with higher levels of replication is needed to better understand long-term impacts and acid-base regulatory mechanisms in this important pelagic fish.

Pharmacokinetic Alterations Associated with Critical Illness.

Haemodynamic, metabolic, and biochemical derangements in critically ill patients affect drug pharmacokinetics and pharmacodynamics making dose optimisation particularly challenging. Appropriate therapeutic dosing depends on the knowledge of the physiologic changes caused by the patient's comorbidities, underlying disease, resuscitation strategies, and polypharmacy. Critical illness will result in altered drug protein binding, ionisation, and volume of distribution; it will also decrease oral drug absorption, intestinal and hepatic metabolism, and renal clearance. In contrast, the resuscitation strategies and the use of vasoactive drugs may oppose these effects by leading to a hyperdynamic state that will increase blood flow towards the major organs including the brain, heart, kidneys, and liver, with the subsequent increase of drug hepatic metabolism and renal excretion. Metabolism is the main mechanism for drug clearance and is one of the main pharmacokinetic processes affected; it is influenced by patient-specific factors, such as comorbidities and genetics; therapeutic-specific factors, including drug characteristics and interactions; and disease-specific factors, like organ dysfunction. Moreover, organ support such as mechanical ventilation, renal replacement therapy, and extracorporeal membrane oxygenation may contribute to both inter- and intra-patient variability of drug pharmacokinetics. The combination of these competing factors makes it difficult to predict drug response in critically ill patients. Pharmacotherapy targeted to therapeutic goals and therapeutic drug monitoring is currently the best option for the safe care of the critically ill. The aim of this paper is to review the alterations in drug pharmacokinetics associated with critical illness and to summarise the available evidence.

Inhibition of SGLT2 co-transporter by dapagliflozin ameliorates tubular proteinuria and tubule-interstitial injury at the early stage of diabetic kidney disease

Diabetic kidney disease (DKD) is characterized by progressive impairment of kidney function. It has been postulated that tubule-interstitial injury, associated with tubular albuminuria, precedes glomerular damage in the early stage of DKD. Here, we wanted to determine if the development of tubule-interstitial injury at the early stage of DKD implies modulation of megalin-mediated protein reabsorption in proximal tubule epithelial cells (PTECs) by SGLT2-dependent high glucose influx. Rats with streptozotocin (STZ)-induced diabetes were treated or not with dapagliflozin (DAPA) for 8 weeks. Four experimental groups were generated: (1) CONT, control; (2) DAPA, rats treated with DAPA; (3) STZ, diabetic rats; (4) STZ + DAPA, diabetic rats treated with DAPA. No changes in glomerular structure and function were observed. The STZ group presented proteinuria and albuminuria associated with an increase in the fractional excretion of proteins. A positive correlation between glycemia and proteinuria was found. These phenomena were linked to a decrease in luminal and total megalin expression and, consequently, in albumin reabsorption in PTECs. We also observed tubule-interstitial injury characterized by an increase in urinary tubular injury biomarkers and changes in tubular histomorphometry parameters. In addition, inverse correlations were found between cortical albumin uptake and tubule-interstitial injury or glycemia. All these modifications were attenuated in the STZ + DAPA group. These results suggest that SGLT2-dependent high glucose influx into PTECs promotes a harmful effect on the PTECs, leading to the development of tubular albuminuria and tubule-interstitial injury preceding glomerular damage. These results expand current knowledge on the renoprotective effects of gliflozins.

Effectiveness and safety of the herbal preparation Soveren® in patients with hyperuricemia

Background. Despite advances in the field of synthetic pharmaceuticals, interest in the use of natural raw materials from representatives of the flora persists throughout the world. Currently, herbal medicine is successfully used in the treatment of both hyperuricemia (HU) and gout.&#x0D; Aim. To evaluate the effectiveness and safety of the herbal preparation Soveren in the prevention and treatment of HU.&#x0D; Materials and methods. The study included 80 patients who were divided into 2 groups: group 1 consisted of patients diagnosed with arthritis with HU (n=40), group 2 patients with gout (n=40). The natural complex of natural origin Soveren was prescribed according to the standard scheme against the background of dietary recommendations. Patients with gout also received urate-lowering therapy. Clinical efficacy and safety of treatment were assessed after 6 and 12 weeks according to the dynamics of laboratory parameters of blood and urine, the EuroQol-5D quality of life questionnaire, assessment of the health status of patients, assessment of patient satisfaction with treatment.&#x0D; Results. The use of the plant complex Soveren in patients with HU does not have a negative effect on the functional state of the liver and kidneys. The drug has a significant positive effect on blood levels, reducing uric acid by 1.5 times, creatinine by 1.11 times, and urea by 1.3 times. In group 2, the addition of the Soveren complex made it possible to achieve the target value of uric acid. In addition, in patients of both groups, within 3 months, the level of total protein in the urine decreased by 3 times, a decrease in the degree of leukocyturia was noted. Against the background of taking the drug in both groups, the relative density of urine increases within the normal range, and a slight acidification of urine was revealed, as well as an increase in urinary excretion of ketones. Most patients positively assessed the result of treatment.&#x0D; Conclusion. Our data confirm the good therapeutic effect and safety of the herbal complex Soveren in patients with HU. With the introduction of the Soveren drug into practice, doctors will have a new effective tool for monitoring the course of HU.

Health Risk in a Geographic Area of Thailand with Endemic Cadmium Contamination: Focus on Albuminuria.

An increased level of cadmium (Cd) in food crops, especially rice is concerning because rice is a staple food for over half of the world’s population. In some regions, rice contributes to more than 50% of the total Cd intake. Low environmental exposure to Cd has been linked to an increase in albumin excretion to 30 mg/g creatinine, termed albuminuria, and a progressive reduction in the estimated glomerular filtration rate (eGFR) to below 60 mL/min/1.73 m2, termed reduced eGFR. However, research into albuminuria in high exposure conditions is limited. Here, we applied benchmark dose (BMD) analysis to the relevant data recorded for the residents of a Cd contamination area and a low-exposure control area. We normalized the excretion rates of Cd (ECd) and albumin (Ealb) to creatinine clearance (Ccr) as ECd/Ccr and Ealb/Ccr to correct for differences among subjects in the number of surviving nephrons. For the first time, we defined the excretion levels of Cd associated with clinically relevant adverse kidney health outcomes. Ealb/Ccr varied directly with ECd/Ccr (β = 0.239, p < 0.001), and age (β = 0.203, p < 0.001), while normotension was associated with lower Ealb/Ccr (β = −0.106, p = 0.009). ECd/Ccr values between 16.5 and 35.5 ng/L of the filtrate were associated with a 10% prevalence of albuminuria, while the ECd/Ccr value of 59 ng/L of the filtrate was associated with a 10% prevalence of reduced eGFR. Thus, increased albumin excretion and eGFR reduction appeared to occur at low body burdens, and they should form toxicity endpoints suitable for the calculation of health risk due to the Cd contamination of food chains.

Polymeric Nanomicelles Loaded with Anandamide and Their Renal Effects as a Therapeutic Alternative for Hypertension Treatment by Passive Targeting.

We have previously demonstrated significant in vitro natriuretic effects of anandamide (AEA) nanoformulation in polymeric nanoparticles, whose size prevents their accumulation in organs, such as the kidneys. Therefore, it is of particular interest to design and test nanostructures that can pharmacologically accumulate in these organs. In this regard, we prepared and characterized polymeric nanomicelles (~14 and 40 nm). Likewise, their biodistribution was determined. Spontaneously hypertensive rats (SHR) and normotensive rats (WKY), n = 3 per group, were divided into five treatment conditions: control, sham, free AEA freshly dispersed in aqueous solution or 24 h after its dispersion, and AEA encapsulated in nanomicelles. The kidneys were the main site of accumulation of the nanoformulation after 24 h. Freshly dispersed free AEA showed its classical triphasic response in SHR, which was absent from all other treatments. Nanoformulated AEA produced a sustained antihypertensive effect over 2 h, accompanied by a significant increase in fractional sodium excretion (FSE %). These effects were not observed in WKY, sham, or free AEA-treated rats after 24 h of its aqueous dispersion. Without precedent, we demonstrate in vivo natriuretic, diuretic, and hypotensive effects of AEA nanoformulation in polymeric nanomicelles, suggesting its possible use as a new antihypertensive agent with intravenous administration and passive renal accumulation.

Iron requirements of broiler chickens as affected by supplemental phytase.

Iron is routinely supplemented in broiler feeds intending to prevent dietary deficiencies. The present research was conducted with the objective of assessing Fe requirements of broilers when fed supplemental phytase. A total of 1,280 1-d-old male Cobb × Cobb 500 were distributed in a 2 × 5 factorial arrangement (phytase-supplemented feeds × 5 graded increases of supplemental Fe) in 80 battery cages, eight replications of eight chicks each. The trial was replicated once. Chicks were fed a Fe-deficient diet without phytase (Fe analyzed at 31.30 ± 3.79 mg/kg) from placement to 7 d and then randomly distributed into battery cages with corresponding dieting treatments with or without phytase and graded increases of supplemental Fe. Feeds were formulated with corn and soybean meal (SBM), laboratory-grade calcium carbonate, and phosphoric acid; therefore, the vast majority of dietary Fe originated from corn and SBM (analyzed diet had 53.3 ± 1.41 mg/kg Fe). Phytase was added in excess to the producer recommendation of 1,000 FYT (4,452 ± 487 FYT/kg analyzed) such that phytate degradation was expected to be maximized. Supplemental Fe was from laboratory-grade ferrous sulfate heptahydrate (FeSO47H2O) which was increasingly added to the feeds (analyzed Fe in the supplemented feeds were: 53.3 ± 1.41, 65.5 ± 0.59, 77.2 ± 1.97, 87.6 ± 1.72, 97.7 ± 1.33 mg/kg). There were no interactions between phytase and dietary Fe for any response throughout the study (P > 0.05). Supplementing phytase had no effects on Fe intake or Fe excretion, as well as on hematocrit (Ht), hemoglobin (Hb), ferritin, Fe contents in the liver or thigh muscle color (P > 0.05). However, phytase-supplemented feeds produced better live performance as well as higher ileal digestible energy and Fe digestibility (P < 0.05). No effects were found for dietary Fe in live performance at day 28 (P > 0.05). On the other hand, increasing dietary Fe led to linear increases in Fe retention and excretion, Fe contents in livers, as well as Ht and Hb at 14 d (P < 0.05). Quadratic responses (P < 0.05) were observed for Hb at 21 d, serum ferritin on days 14, 21, and 28 (maximum responses were 83.3, 104.0, 91.9, and 88.3 mg/kg Fe, respectively). In conclusion, supplementing Fe adding to a total of 97.7 mg/kg dietary Fe did not affect live performance traits. However, the average of Fe-related blood parameters was maximized at 91.9 mg/kg dietary Fe. Supplementing phytase provided a significant increase in Fe digestibility.

S-16-4: PHARMACOKINETICS, PHARMACODYNAMICS, AND SAFETY RESULTS FROM A PHASE 1 MULTIPLE ASCENDING DOSE STUDY OF THE SELECTIVE ALDOSTERONE SYNTHASE INHIBITOR CIN-107

Objective: To evaluate the safety, pharmacokinetics, and pharmacodynamics of multiple ascending doses of the highly potent, selective small-molecule inhibitor of aldosterone synthase (CIN-107) in a randomized, double-blind, placebo-controlled phase 1 study in healthy volunteers. Design and methods: Subjects were randomized into 5 cohorts to receive CIN-107 or placebo once daily for 10 days. Cohorts 1 and 2 were placed on a low salt diet to stimulate aldosterone production and were administered 2.5 or 5.0 mg oral CIN-107, respectively. They also underwent an adrenocorticotropic hormone challenge to increase aldosterone and cortisol levels to evaluate the specificity of CIN-107 for aldosterone synthase. Cohorts 3, 4, and 5 were placed on a normal salt diet and were administered 1.5, 2.5, or 0.5 mg oral CIN-107, respectively. Blood samples were collected prior to and after dosing on days 1 and 10 for measurement of plasma CIN-107 concentrations to characterize single-dose and steady-state pharmacokinetics. Pharmacodynamic measurements included aldosterone, cortisol, and electrolytes. Safety assessments included physical examination, electrocardiograms, orthostatic vital signs, and clinical laboratory evaluations, including renal function. Results: Fifty-four subjects completed the study. There were no deaths, serious adverse events, or discontinuations due to treatment-emergent adverse events (TEAEs). All TEAEs in subjects receiving CIN-107 were mild in severity. Plasma concentrations of CIN-107 increased proportionally with ascending doses. CIN-107 was rapidly absorbed, with peak concentrations observed within 4 hours after dosing. Mean half-life was 26–31 hours. A dose-dependent reduction of plasma aldosterone occurred with CIN-107 doses ≧1.5 mg, regardless of normal or low salt diet. Decreases in plasma aldosterone started on Day 1 and were sustained, with levels reduced by approximately 51–73% on Day 10 (Figure 1). CIN-107 had no meaningful impact on plasma cortisol levels. The urine sodium:potassium ratio increased on Day 1 and diminished by Day 10, an effect mediated by greater elimination of sodium on Day 1 compared to Day 10. A mild increase in the blood urea nitrogen:creatinine ratio and a mild reduction in the glomerular filtration rate (&lt;15%) suggest that CIN-107 produced a mild diuretic effect. Conclusions: CIN-107 was safe and well tolerated and resulted in dose-dependent increases in plasma CIN-107 with a half-life that supports once-daily dosing. The dose-dependent decrease in plasma aldosterone, transient increase in sodium excretion, and mild diuretic effect support continued study in ongoing phase 2 clinical trials evaluating the efficacy and safety of CIN-107 for treatment-resistant or uncontrolled hypertension and primary aldosteronism.

Interaction between inorganic phosphate concentration and glucose metabolism in mild refeeding syndrome model.

Refeeding syndrome is a major clinical problem that leads to fatal complications in patients suffering from malnutrition. Hypophosphatemia inevitably is observed at the onset of refeeding syndrome and therefore is monitored during refeeding; however, the causes of metabolic changes in phosphate concentration during refeeding remain poorly understood. In a previous study, we established a refeeding syndrome model employing total parenteral nutrition with insulin-induced hypophosphatemia, but the symptoms were severe and the metabolic mechanisms in this model may not have been representative of clinical conditions. Therefore, we established a new animal model of mild refeeding syndrome by using a shorter fasting period followed by a single refeeding. These mild refeeding syndrome-model rats exhibited hypophosphatemia without increases in urinary phosphate excretion. Interestingly, administration of the combination of phosphate and insulin during refeeding promoted insulin secretion during refeeding. This model implies that Pi may directly promote insulin secretion in pancreatic cells. These results clarify the interaction between phosphate and glucose metabolism pancreatic cells during refeeding syndrome in a mild refeeding syndrome model.

ВОЗМОЖНОСТИ ПРИМЕНЕНИЯ БАЛЬНЕОТЕРАПИИ В КОМПЛЕКСНОМ ЛЕЧЕНИИ ГИПЕРКАЛЬЦИУРИИ У ДЕТЕЙ

The annual increase in the number of children with urolithiasis dictates the need to develop an integrated approach to prevent the development of this pathology. The most common metabolic disorder that can lead to the development of KSD is idiopathic hypercalciuria (IG). One of the main components for the correction of hypercalciuria is an adequate drinking regimen, including low-mineralized and slightly alkaline mineral waters. The aim of the study was to evaluate the effectiveness of Silver Key mineral water usage in the complex therapy of IG in children. Materials and methods. The study included 50 children with IG aged 3 to 17 years who underwent a study of clinical and anamnestic data, a laboratory study of blood and urine to assess indicators of phosphorus-calcium metabolism, and an ultrasound examination of the urinary system. During the study, two groups of children with IG were formed. The children of the study group (n=30) received Silver Key mineral water in the complex therapy of IG, the children of the control group (n=20) received ordinary drinking water as a drinking regimen. After therapy in groups of children, a comparative evaluation of effectiveness of the therapy was carried out according to clinical and laboratory parameters. Results. We found that in the children of the study group, in the background of Silver Key mineral water in the complex therapy, there was a positive trend that exceeded the effectiveness of the treatment of patients in the control group. The percentage of children who consumed mineral water, in which normalization of clinical and laboratory manifestations of IG was recorded, turned out to be significantly higher. Conclusion. The addition of Silver Key mineral water to the IG treatment regimen significantly reduces the increase in urinary calcium excretion and helps to reduce the severity of clinical manifestations of IG in children, allowing us to recommend its use for correction of IG and prevention of urolithiasis in children.

Novel mutation on SLC5A2 gene causing familial renal glucosuria in a patient with hypokalemic periodic paralysis type 2.

Introduction: The presence of glucosuria is usually a red flag that leads to diabetes mellitus screening in pediatric patients. Despite this correct approach, high levels of glucosuria without hyperglycemia should raise suspicion for other diagnoses, namely renal tubular disease, such as Familial Renal Glucosuria (FRG). A large number of mutations on the solute carrier family 5 member 2 (SLC5A2) gene, which encodes sodium-glucose cotransporter 2 (SGLT2), have been reported to be responsible for the increase in renal excretion of glucose.(1) Case Report: We report a case of a teenager, followed in our outpatient clinic for Hypokalemic Periodic Paralysis type 2 (HyPP2), in which persistent renal glucosuria was fortuitously found. Genetic testing for SLC5A2 gene was conducted, leading to the identification of two mutations in the SGLT2 gene, both in heterozygosity, one in exon 14 (N654S), previously reported, and another in exon 2 (A94C), a novel mutation not yet described in population databases. Discussion: The fact that our patient had mutations in two different genes from different chromosomes, both related to sodium channels and one of these not previously described, makes this case interesting. The key point is that it is important to be aware of these diseases and assess the child as a whole. Despite the apparent benignity of these two diseases, clinical surveillance must be tight. We advise avoiding prolonged fasting, a diet rich in carbohydrates and intense physical exercise. Conclusion: In the presence of isolated glucosuria, without hyperglycemia, we should suspect Familial Renal Glucosuria. Familial Renal Glucosuria is a rare disease caused by mutation in SGLT2 gene. Exon 2 (A94C) in heterozygosity is a novel mutation not yet described in population databases. This is the first description of a patient with both Familial Renal Glucosuria and Hipokalemic periodic paralysis type 2. The association of both these diseases had not yet been reported.

Rare causes of Hypophosphatemia: diagnostic approach

Hypophosphatemia is common and may be overlooked due to its asymptomatic nature or non-specific symptoms. Two main mechanisms are at its origin: a shift towards the intracellular sector and an increase in urinary phosphate excretion. A measurement of the urinary phosphate reabsorption threshold allows a diagnostic orientation. Alongside common forms of parathyroid hormone-dependent hypophosphatemia, one should not ignore rare FGF23-mediated forms, in particular X-linked hypophosphatemic rickets. The treatment, above all etiological, also includes the administration of phosphate and, in the event of an excess of FGF23, supplementation with calcitriol. In cases of oncogenic osteomalacia and X-linked hypophosphatemic rickets, the use of burosumab, an anti-FGF23 antibody, must be considered.

Ergosterol fraction from Agaricus bisporus modulates adipogenesis and skeletal glucose uptake in high fat diet induced obese C57BL/6 mice

AimThe study aimed to optimize a method of extracting ergosterol rich concentrate (ECF) and to evaluate its significant impact on adipogenesis and associated complications in high-fat diet (HFD) induced obese mice. MethodsA comparative analysis (soxhlet and ultra sound assisted extraction) was done to obtain the highest yield of ergosterol from Agaricus bisporus. The ECF was evaluated for the biological effect on 3T3-L1 pre-adipocytes in-vitro and in male C57BL/6 mice model in-vivo. Key findingsUltra sound assisted extraction method using the solvent n-hexane resulted in highest ergosterol yield. ECF treatment significantly reduced the differentiation and lipid accumulation in pre-adipocyte cells without any cytotoxicity. In-vivo study illustrated beneficial impact on cholesterol metabolism by down regulating the hepatic gene expression of LXR-α, HMG-CoR and up-regulating LDL-R expression. Significant increase in fecal excretion of cholesterol and bile acids have also been observed among the ECF treated animals compared to high fat diet (HFD) fed mice. ECF had an anti-adipogenic activity in-vivo mainly by inhibiting the activity of PPAR-γ, C/EBP-α and SREBP-1c. The results also depicted the improvement of obesity associated insulin resistance by ECF treatment manly via decrease in plasma resistin and up-regulation in skeletal GLUT4 protein expression. SignificanceOur study illustrated diverse activity of ECF in the therapeutic management of obesity associated metabolic complications mainly by reducing adipogenesis and improving glucose uptake in skeletal muscle in conjunction with improved cholesterol metabolism.