1,4‐Dihydropyridine derivatives without Ca2+‐antagonist activity up‐regulate Psma6 mRNA expression in kidneys of intact and diabetic rats

Impaired degradation of proteins by the ubiquitin–proteasome system (UPS) is observed in numerous pathologies including diabetes mellitus (DM) and its complications. Dysregulation of proteasomal degradation might be because of altered expression of genes and proteins involved in the UPS. The search for novel compounds able to normalize expression of the UPS appears to be a topical problem. A novel group of 1,4‐dihydropyridine (1,4‐DHP) derivatives lacking Ca2+‐antagonists activities, but capable to produce antidiabetic, antioxidant and DNA repair enhancing effects, were tested for ability to modify Psma6 mRNA expression levels in rat kidneys and blood in healthy animals and in rats with streptozotocin (STZ) induced DM. Psma6 gene was chosen for the study, as polymorphisms of its human analogue are associated with DM and cardiovascular diseases. 1,4‐DHP derivatives (metcarbatone, etcarbatone, glutapyrone, J‐9‐125 and AV‐153‐Na) were administered per os for three days (0.05 mg/kg and/or 0.5 mg/kg). Psma6 gene expression levels were evaluated by quantitative PCR. Psma6 expression was higher in kidneys compared to blood. Induction of diabetes caused increase of Psma6 expression in kidneys, although it was not changed in blood. Several 1,4‐DHP derivatives increased expression of the gene both in kidneys and blood of control and model animals, but greater impact was observed in kidneys. The observed effect might reflect coupling of antioxidant and proteolysis‐promoting activities of the compounds. Copyright © 2015 John Wiley & Sons, Ltd.     

Compromission de la masse osseuse et de la motricité chez des malades traités par blocage androgénique

Therapeutic androgen suppression induces hypogonadism with effects on the patient’s locomotor system. We tried to verify these effects on a group of patients with prostate cancer presenting a prolonged life expectancy. Thirty six patients treated by radical prostatectomy (mean PSA: 7.2±1.3 ng/ml) had stage pT3 cancer in 24 cases and pT2c in 12 cases. The first group was treated by radiotherapy and androgen suppression and the second group was treated by androgen suppression alone after surgery. After 24–36 months (mean=28.4 months), staging was performed by CT scan, bone scintigraphy, PSA and testosterone assays, and bone densitometry. An identical assessment was repeated an average of 53.1 months after starting treatment. Staging never demonstrated disease recurrence; PSA was between 0.01 and 0.4 ng/ml (mean: 0.11 ± 0.96 ng/ml) and the mean plasma testosterone was 0.4 ng/ml. The first bone densitometry revealed osteopenia: T score =?1.71±0.91; Ward score=?2.22±0.917; BMD (bone density) =0.879±0.126. The second bone densitometry showed progression to osteoporosis and a significant 6% reduction of the BMD: T Score=?1.95±0.84; Ward score=?2.4±0.87; BMD=0.819±0.12. During this time interval, 3 patients developed a fracture of the femur and a fourth patient fractured two ribs after physical exertion. All patients compalined of decreased physical strength and very marked fatigability. We can conclude that androgen suppression causes an alteration of locomotor function and quality of life of patients treated for prostate cancer and presenting a long life expectancy.     

Transgenic Fatal Familial Insomnia Mice Indicate Prion Infectivity-Independent Mechanisms of Pathogenesis and Phenotypic Expression of Disease

Fatal familial insomnia (FFI) and a genetic form of Creutzfeldt-Jakob disease (CJD¹⁷⁸) are clinically different prion disorders linked to the D178N prion protein (PrP) mutation. The disease phenotype is determined by the 129 M/V polymorphism on the mutant allele, which is thought to influence D178N PrP misfolding, leading to the formation of distinctive prion strains with specific neurotoxic properties. However, the mechanism by which misfolded variants of mutant PrP cause different diseases is not known. We generated transgenic (Tg) mice expressing the mouse PrP homolog of the FFI mutation. These mice synthesize a misfolded form of mutant PrP in their brains and develop a neurological illness with severe sleep disruption, highly reminiscent of FFI and different from that of analogously generated Tg(CJD) mice modeling CJD¹⁷⁸. No prion infectivity was detectable in Tg(FFI) and Tg(CJD) brains by bioassay or protein misfolding cyclic amplification, indicating that mutant PrP has disease-encoding properties that do not depend on its ability to propagate its misfolded conformation. Tg(FFI) and Tg(CJD) neurons have different patterns of intracellular PrP accumulation associated with distinct morphological abnormalities of the endoplasmic reticulum and Golgi, suggesting that mutation-specific alterations of secretory transport may contribute to the disease phenotype.     

Characterization of NAD:arginine ADP-ribosyltransferases

NAD:arginine mono-ADP-ribosyltransferases catalyze the transfer of ADP-ribose from NAD to the guanidino group of arginine on a target protein. Deduced amino acid sequences of one family (ART1) of mammalian ADP-ribosyltransferases, cloned from muscle and lymphocytes, show hydrophobic amino and carboxyl termini consistent with glycosylphosphatidylinositol (GPI)-anchored proteins. The proteins, overexpressed in mammalian cells transfected with the transferase cDNAs, are released from the cell surface with phosphatidylinositol-specific phospholipase C (PI-PLC), and display immunological and biochemical characteristics consistent with a cell surface, GPI-anchored protein. In contrast, the deduced amino acid sequence of a second family (ART5) of transferases, cloned from murine lymphoma cells and expressed in high abundance in testis, displays a hydrophobic amino terminus, consistent with a signal sequence, but lacks a hydrophobic signal sequence at its carboxyl terminus, suggesting that the protein is destined for export. Consistent with the surface localization of the GPI-linked transferases, multiple surface substrates have been identified in myotubes and activated lymphocytes, and, notably, include integrin subunits. Similar to the bacterial toxin ADP-ribosyltransferases, the mammalian transferases contain the characteristic domains involved in NAD binding and ADP-ribose transfer, including a highly acidic region near the carboxy terminus, which, when disrupted by in vitro mutagenesis, results in a loss of enzymatic activity. The carboxyl half of the protein, synthesized as a fusion protein in E. coli, possessed NADase, but not ADP-ribosyltransferase activity. These findings are consistent with the existence at the carboxyl terminus of ART1 of a catalytically active domain, capable of hydrolyzing NAD, but not of transferring ADP-ribose to a guanidino acceptor.     

A mathematical description of blood spiral flow in vessels: application to a numerical study of flow in arterial bending

Local arterial haemodynamics has been associated with the pathophysiology of several cardiovascular diseases. The stable spiral blood-flows that were observed in vivo in several vessels, may play a dual role in vascular haemodynamics, beneficial since it induces stability, reducing turbulence in the arterial tree, and accounts for normal organ perfusion, but detrimental in view of the imparted tangential velocities that are involved in plaque formation and development. Being a spiral flow considered representative of the local blood dynamics in certain vessels, a method is proposed to quantify the spiral structure of blood flow. The proposed function, computed along a cluster of particle trajectories, has been tested for the quantitative determination of the spiral blood flow in a three-dimensional, s-shaped femoral artery numerical model in which three degrees of stenosis were simulated in a site prone to atherosclerotic development. Our results confirm the efficacy of the Lagrangian analysis as a tool for vascular blood dynamics investigation. The proposed method quantified spiral motion, and revealed the progression in the degree of stenosis, in the presented case study. In the future, it could be used as a synthetic tool to approach specific clinical complications.     

Intraperitoneal insulin in uraemic diabetics undergoing continuous ambulatory peritoneal dialysis.

The kinetics of absorption of intraperitoneally administered insulin were studied in nine uraemic insulin-dependent diabetics undergoing continuous ambulatory peritoneal dialysis (CAPD). In each of three studies 20 U of regular insulin was directly injected as a bolus into the peritoneal cavity through an indwelling Tenckhoff catheter. In two procedures the insulin injection was followed by the instillation of either 2 litres of 1.5% dextrose dialysates or 2 litres of 4.5% dextrose dialysate. In the third 20 ml of saline was used to flush the tubing. Plasma free insulin values rose more rapidly and reached significantly higher concentrations (55.6 +/- 18.8 mU/l) when the insulin had been injected into an empty peritoneal cavity than when it was followed by dialysate. These differences were observed despite the fact that most of the insulin injected was retained by the patients. Since the plasma insulin values did not differ after instillations of dialysate containing 1.5% and 4.5% dextrose, the osmolality of the dialysate seemed not to affect insulin absorption, and the dilution of the insulin probably delayed its transfer through the peritoneum. These findings suggest that insulin given intraperitoneally to patients undergoing CAPD will be most effective if it is given into an empty peritoneal cavity at least 30 minutes before the dialysate is instilled.