Absence of interferons-alpha and -gamma in renal lesions of systemic lupus erythematosus and membranous glomerulonephritis

Frozen kidney biopsy sections from nine patients with systemic lupus erythematosus (SLE) as well as many other renal diseases, including IgA nephropathy, membranous nephritis, and minimal change nephrotic syndrome, were negative for interferons -alpha and -gamma by immunofluorescence. Lupus patients studied included several subjects with marked serum elevations of interferon activity as well as others with low or negative serum interferon levels. Isolated glomerular eluates prepared from normal and SLE kidneys showed no functional interferon activity by virus plaque inhibition assay. Components of normal as well as SLE serum showed no direct binding to interferon -alpha or -gamma by ELISA assays.     

Clonidine suppression and its adrenoreceptor mediation in schedule-induced polydipsia

Rats were trained in a fixed-interval, one-minute (FI 1 min) food reinforcement schedule for 1 hour daily at reduced body weight until their lever presses, licks and water intake all became stabilized for 6 days. Two experiments were performed to examine the function of sympathetic activity in schedule-induced polydipsia. In experiment 1, intracerebroventricular injection of clonidine (0.75-37.5 nmol) produced a dose-related suppression of schedule-induced drinking and licking and schedule-dependent lever pressing; these effects were later attenuated by yohimbine (5 nmol) pretreatment. Prazosin (10 nmol) also decreased clonidine-induced suppression of lever pressing, whereas neither prazosin (10 nmol) nor naloxone (10 nmol) caused any alteration in the suppression effects of clonidine on drinking and licking. None of these antagonists alone changed an individual rat's preestablished behavioral baselines. In experiment 2, the endogenous catecholamine levels, were determined in frontal cortex, hypothalamus, brainstem, dorsal obex area and adrenal glands. During the SIP situation, both the epinephrine level in adrenal glands and the norepinephrine level in hypothalamus were elevated.     

Biochemical characterization of human Thy-1

The human Thy-1 homologue (p25) was characterized biochemically for amino acid composition, sequence and carbohydrate content. Two other forms of the human Thy-1 molecule were detected and partially characterized. A 40,000 mol.wt. molecule (p40) is the dimer of p25 and its formation is increased by the presence of sodium dodecyl sulphate (SDS). The second form of 16,000 mol.wt. (p16) appears to be a cryptic or breakdown form of p25. Comparison of the amino acid compositions of p25, p40 and p16 isolated from MOLT-3 cells, with that deduced from the nucleotide sequence of the gene coding for part of the putative T cell antigen receptor, also from MOLT-3 cells, shows that the Thy-1 homologue is distinct from, but evolutionary related to, one of the putative T cell antigen receptor polypeptide chains.     

Topography and behavior of Sertoli cells in sparse culture during the transitional remodeling phase

We report observations on the behavior of Sertoli cells in sparse culture during the period from the time of plating to the time of initial confluence (the transitional remodeling phase). Changes in shape, structure, and polarity of cells, as well as changes in migration patterns and cell-cell association patterns, have been followed during the transitional remodeling phase with the aid of topographical markers. These markers are based upon differences between ultrastructural features of the basolateral and apicolateral surfaces. The basolateral surface is characterized by plasmalemmal blebs, whereas the apicolateral surface is characterized by filopodial extensions. Structural differences observed in situ remain evident in Sertoli cells isolated by sequential enzymatic treatments that are described. Another marker is provided by laminin-binding sites, which are detected exclusively on the blebbed, basolateral surfaces of freshly prepared Sertoli cell aggregates. The orientation described is sustained during the initial radial migration of Sertoli cells explanted on uncoated glass coverslips. Under these conditions, blebs are detected only on the dorsal surfaces, and filopodial extensions are evident only on the ventral surfaces. In contrast, Sertoli cells sparsely plated on a reconstituted basement membrane (air-dried Matrigel) migrate rapidly, display an extraordinary capacity to form elaborate cytoplasmic extensions for cell-cell and cell-substratum contacts, and readily retract blebs and filopodial extensions. These cells do not form mosaic borders, whereas cells plated on uncoated glass do form a monolayer with mosaic-like borders. Cells sparsely seeded on gelated Matrigel migrate preferentially at gaps between adjacent cell explants, and develop a compact cell-cell association pattern. These cells display few, if any, cytoplasmic extensions. We compare the behavior of Sertoli cells sparsely plated on Matrigel with the behavior of Sertoli cells in situ during different stages of development.     

Effect of hypothermia on the intestinal absorption of uracil and L-dopa in the rat

The in situ rat gut technique was used to explore the effects of hypothermia on the intestinal absorption of L-dopa and uracil. A hypothermic state was induced when male Sprague-Dawley rats, weighing between 300 and 370 g, were exposed to an atmosphere of helox (helium:oxygen, 80:20) at 0-4 degrees C. After 4 to 5 h, the rectal temperatures were decreased from 37 to 20 degrees C. The rewarming process for hypothermic animals undergoing anesthesia appears to be prolonged. The animals were prepared for surgery using ether as anesthetic after a rectal temperature of 20 degrees C was attained. Hypothermia showed a significant influence on decreasing L-dopa and uracil disappearances from the intestinal lumen. About 40 percent reduction of the rates of disappearance was observed with a 10 degrees C reduction of the rectal temperature. Additionally, a rapid drop of the rate of drug disappearance was observed in the beginning stage of rectal temperature decrease (31-36 degrees C) as well as in the hypothermic state with rectal temperatures < 26 degrees C. There appears to be a thermal stable region for absorption between 26 and 30 degrees C. Water efflux was studied in both normothermic and hypothermic animals. Normothermic rats showed greater average cumulative water efflux over a period of 30 min (71 and 61% for L-dopa and uracil, respectively) in comparison with that of hypothermic rats (34 and 41% for L-dopa and uracil, respectively). The cold-treated animals showed decreased rates of disappearance associated with the decreased water efflux. Therefore, the reduction of the rates of drug disappearance from the intestinal lumen caused by hypothermia may be partially related to the decrease of water efflux during hypothermia.     

Insulinemic and Inflammatory Dietary Patterns and Risk of Prostate Cancer

BackgroundHyperinsulinemia and inflammation are inter-related pathways that link diet with the risk of several chronic diseases. Evidence suggests that these pathways may also increase prostate cancer risk.ObjectiveTo determine whether hyperinsulinemic diet and inflammatory diet are associated with prostate cancer incidence and mortality.Design, setting, and participantsWe prospectively followed 41 209 men in the Health Professionals Follow-up Study (1986–2014). Scores for two validated dietary patterns were calculated from food frequency questionnaires at baseline and updated every 4 yr.Outcome measurements and statistical analysisTotal, advanced, and lethal prostate cancer outcomes were assessed. Multivariable-adjusted hazard ratios (HRs) and 95% confidence intervals (CIs) were determined for associations between two empirical hypothesis-oriented dietary patterns—empirical dietary index for hyperinsulinemia and empirical dietary inflammatory pattern—and prostate cancer risk estimated using Cox proportional hazard regression.Results and limitationsDuring 28 yr of follow-up, 5929 incident cases of total prostate cancer, including 1019 advanced and 667 fatal, were documented. In multivariable-adjusted models, there was a 7% higher risk of advanced prostate cancer (HR: 1.07; 95% CI: 1.01–1.15) and a 9% higher risk of fatal prostate cancer (HR: 1.09; 95% CI: 1.00–1.18) per standard deviation (SD) increase in the hyperinsulinemic diet. When stratified by age, the hyperinsulinemic diet was associated with only earlier-onset aggressive prostate cancer (men under 65 yr), with per SD HRs of 1.20 (95% CI: 1.06–1.35) for advanced, 1.22 (1.04–1.42) for fatal, and 1.20 (1.04–1.38) for lethal. The inflammatory diet was not associated with prostate cancer risk in the overall study population, but was associated with earlier-onset lethal prostate cancer (per SD increase HR: 1.16; 95% CI: 1.00–1.35).ConclusionsHyperinsulinemia and inflammation may be potential mechanisms linking dietary patterns with the risk of aggressive prostate cancer, particularly earlier-onset disease.Patient summaryAvoiding inflammatory and hyperinsulinemic dietary patterns may be beneficial for the prevention of clinically relevant prostate cancer, especially among younger men.     

Fifty years of the mitochondrial pyruvate carrier: New insights into its structure,function, and inhibition

The mitochondrial pyruvate carrier (MPC) resides in the mitochondrial inner membrane, where it links cytosolic and mitochondrial metabolism by transporting pyruvate produced in glycolysis into the mitochondrial matrix. Due to its central metabolic role, it has been proposed as a potential drug target for diabetes, non-alcoholic fatty liver disease, neurodegeneration, and cancers relying on mitochondrial metabolism. Little is known about the structure and mechanism of MPC, as the proteins involved were only identified a decade ago and technical difficulties concerning their purification and stability have hindered progress in functional and structural analyses. The functional unit of MPC is a hetero-dimer comprising two small homologous membrane proteins, MPC1/MPC2 in humans, with the alternative complex MPC1L/MPC2 forming in the testis, but MPC proteins are found throughout the tree of life. The predicted topology of each protomer consists of an amphipathic helix followed by three transmembrane helices. An increasing number of inhibitors are being identified, expanding MPC pharmacology and providing insights into the inhibitory mechanism. Here, we provide critical insights on the composition, structure, and function of the complex and we summarize the different classes of small molecule inhibitors and their potential in therapeutics.