Involvement of the Tubular ClC-Type Exchanger ClC-5 in Glomeruli of Human Proteinuric Nephropathies

Glomerular protein handling mechanisms have received much attention in studies of nephrotic syndrome. Histopathological findings in renal biopsies from severely proteinuric patients support the likelihood of protein endocytosis by podocytes. ClC-5 is involved in the endocytosis of albumin in the proximal tubule.

Aim

To investigate whether ClC-5 is expressed in the glomerular compartment and whether it has a role in proteinuric nephropathies. ClC-5 expression was studied using Real-time PCR in manually- and laser-microdissected biopsies from patients with type 2 diabetes (n 37) and IgA nephropathy (n 10); in biopsies of membranous glomerulopathy (MG) (n 14) immunohistochemistry for ClC-5 (with morphometric analysis) and for WT1 was done. Controls: cortical tissue (n 23) obtained from unaffected parts of tumor-related nephrectomy specimens.

Results

ClC-5 was expressed at glomerular level in all biopsies. Glomerular ClC-5 levels were significantly higher in diabetic nephropaty and MG at both mRNA and protein level (p<0.002; p<0.01). ClC-5 and WT1 double-staining analysis in MG showed that ClC-5 was localized in the podocytes. ClC-5 ultrastructural immunolocalization was demonstrated in podocytes foot processes. Our study is the first to demonstrate that ClC-5 is expressed in human podocytes. The ClC-5 overexpression found in biopsies of proteinuric patients suggests that proteinuria may play a part in its expression and that podocytes are likely to have a key role in albumin handling in proteinuric states.     

Right ventricular systolic pressure load alters myocyte maturation in fetal sheep

The effects of right ventricular (RV) systolic pressure (RVSP) load on fetal myocyte size and maturation were studied. Pulmonary artery (PA) pressure was increased by PA occlusion from mean 47.4 +/- 5.0 (+/-SD) to 71 +/- 13.6 mmHg (P < 0.0001) in eight RVSP-loaded near-term fetal sheep for 10 days. The maximal pressure generated by the RV with acute PA occlusion increased after RVSP load: 78 +/- 7 to 101 +/- 15 mmHg (P < 0.005). RVSP-load hearts were heavier (44.7 +/- 8.4 g) than five nonloaded hearts (31.8 +/- 0.2 g; P < 0.03); heart-to-body weight ratio (10.9 +/- 1.1 and 6.5 +/- 0.9 g/kg, respectively; P < 0.0001). RVSP-RV myocytes were longer (101.3 +/- 10.2 microm) than nonloaded RV myocytes (88.2 +/- 8.1 microm; P < 0. 02) and were more often binucleated (82 +/- 13%) than nonloaded myocytes (63 +/- 7%; P < 0.02). RVSP-loaded myocytes had less myofibrillar volume than did nonloaded hearts (44.1 +/- 4.4% and 56. 1 +/- 2.6%; P < 0.002). We conclude that RV systolic load 1) leads to RV myocyte enlargement, 2) has minor effects on left ventricular myocyte size, and 3) stimulates maturation (increased RV myocyte binucleation). Myocyte volume data suggest that RV systolic loading stimulates both hyperplastic and hypertrophic growth.     

L-Carnitine: a potential treatment for blocking apoptosis and preventing skeletal muscle myopathy in heart failure

Skeletal muscle in congestive heartfailure is responsible for increased fatigability and decreasedexercise capacity. A specific myopathy with increased expression offast-type myosins, myocyte atrophy, secondary to myocyteapoptosis triggered by high levels of circulating tumornecrosis factor- (TNF-) has been described. In an animal model ofheart failure, the monocrotaline-treated rat, we have observed anincrease of apoptotic skeletal muscle nuclei. Proapoptoticagents, caspase-3 and -9, were increased, as well as serum levels ofTNF- and its second messenger sphingosine. Treatment of rats withL-carnitine, known for its protective effect on musclemetabolism injuries, was found to inhibit caspases and to decrease thelevels of TNF- and sphingosine, as well as the number ofapoptotic myonuclei. Staurosporine was used in in vitro experimentsto induce apoptosis in skeletal muscle cells in culture. WhenL-carnitine was applied to skeletal muscle cells, before staurosporine treatment, we observed a reduction in apoptosis. These findings show that L-carnitine can preventapoptosis of skeletal muscles cells and has a role in thetreatment of congestive heart failure-associated myopathy.

     

Adhesive recognition between developing retinal cells and the optic tecta of the chick embryo.

An assay is described that can measure the rates of adhesion of isotopically labeled cell bodies from either the dorsal or ventral regions of the chick embryo retina to dorsal and ventral tectal halves. Immediately after dissociation of the retina, both ventral and dorsal retinal cells adhere preferentially to ventral tectal halves. With increasing time after dissociation, however, the preference of the ventral retinal cells shifts to dorsal tectal halves. The dorsal retinal cells continue to show a specificity for ventral tectal halves regardless of the length of time after their dissociation.When the developmental age of the tectal halves is varied from 8–14 days, there is no change in the specificities shown by retinal cells toward these tecta.When retinal age is varied, ventral retinal cells do not adhere preferentially to dorsal tecta until day 6 and older. Dorsal retinas show their specificity toward ventral tecta as early as day three.Control experiments include the use of nonretinal tissues, noninnervated tectal halves and pigmented retinal cells.     

Synthesis of (E)-cinnamyl ester derivatives via a greener Steglich esterification

Cinnamic acid derivatives are known antifungal, antimicrobial, antioxidant, and anticancer compounds. We have developed a facile and mild methodology for the synthesis of (E)-cinnamate derivatives using a modified Steglich esterification of (E)-cinnamic acid. Using acetonitrile as the solvent, rather than the typical chlorinated solvent, and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) as the coupling agent enables ester conversion in 45?min with mild heating (40–45?°C) and an average yield of 70% without need for further purification. These conditions were used to couple (E)-cinnamic acid with 1° and 2° aliphatic alcohols, benzylic and allylic alcohols, and phenols. This work demonstrates a facile and greener methodology for Steglich esterification reactions.     

The hypothalamo-hypophyseal system: the role of calcium ions

The Authors investigated the effects of Verapamil a calcium antagonist substance upon the pituitary response to several secretogogus agents in ten male healthy subjects in order to elucidate the calcium role in the regulation of the hypothalamic pituitary function. As a consequence of Verapamil administration (5 mg iv) a significant hyporesponsiveness of FSH and LH to GnRH (100 micrograms iv) of TSH to TRH (200 gamma iv) and of ACTH to insulin induced hypoglycemia was observed. Prolactin response to TRH (200 gamma iv) was not modified by Verapamil treatment. The Authors concluded emphasizing the importance of Ca++ in the modulation of diencephalic pituitary activity.     

The interleukin-8 (IL-8/CXCL8) receptor inhibitor reparixin improves neurological deficits and reduces long-term inflammation in permanent and transient cerebral ischemia in rats

Leukocyte infiltration is viewed as a pharmacological target in cerebral ischemia. We previously reported that reparixin, a CXCL8 receptor blocker that inhibits neutrophil infiltration, and related molecules can reduce infarct size in a rat model of transient middle cerebral artery occlusion (MCAO). The study aims were to compare the effects of reparixin in transient and permanent MCAO using varied treatment schedules and therapeutic windows to evaluate effects on long-term neurological deficits and late inflammatory response. Reparixin, administered for 1 to 3 days, 3.5 to 6 h after MCAO, ameliorates neurological function recovery and inhibits long-term inflammation. The infarct size reduction at 24 h, evaluated by TTC staining, is more pronounced in transient MCAO. MRI analysis identified a decrease in the progression of infarct size by reparixin that was more evident at 48 h in permanent MCAO, and was associated with a significantly improved recovery from long-term neurological deficits.     

Comparative molecular docking and simulation analysis of molnupiravir and remdesivir with SARS-CoV-2 RNA dependent RNA polymerase (RdRp)

Treatment of SARS-CoV-2 targeting its RNA dependent RNA polymerase (RdRp) is of current interest. Remdesivir has been approved for the treatment of COVID-19 around the world. However, the drug has been linked with pharmacological limitations like adverse effects and reduced efficiency. Nevertheless, recent advancements have depicted molnupiravir as an effective therapeutic agent to target the SARS-CoV-2 RdRp. The drug has cleared both in vitro and in vivo screening. It is in phase-III clinical trial. Nonetheless, there are no data on themolecular binding interaction of molnupiravir with RdRp. Therefore, it is of interest to report the binding interaction of molnupiravir using molecular docking. It is also of interest to show its stability during interaction using molecular dynamics and binding free energy calculations along with drug likeliness and pharmacokinetic properties in comparison with remdesivir.