The anabolic action of intermittent parathyroid hormone on cortical bone depends partly on its ability to induce nitric oxide‐mediated vasorelaxation in BALB/c mice

There is strong evidence that vasodilatory nitric oxide (NO) donors have anabolic effects on bone in humans. Parathyroid hormone (PTH), the only osteoanabolic drug currently approved, is also a vasodilator. We investigated whether the NO synthase inhibitor L‐NAME might alter the effect of PTH on bone by blocking its vasodilatory effect. BALB/c mice received 28 daily injections of PTH[1–34] (80 µg/kg/day) or L‐NAME (30 mg/kg/day), alone or in combination. Hindlimb blood perfusion was measured by laser Doppler imaging. Bone architecture, turnover and mechanical properties in the femur were analysed respectively by micro‐CT, histomorphometry and three‐point bending. PTH increased hindlimb blood flow by >30% within 10 min of injection (P < 0.001). Co‐treatment with L‐NAME blocked the action of PTH on blood flow, whereas L‐NAME alone had no effect. PTH treatment increased femoral cortical bone volume and formation rate by 20% and 110%, respectively (P < 0.001). PTH had no effect on trabecular bone volume in the femoral metaphysis although trabecular thickness and number were increased and decreased by 25%, respectively. Co‐treatment with L‐NAME restricted the PTH‐stimulated increase in cortical bone formation but had no clear‐cut effects in trabecular bone. Co‐treatment with L‐NAME did not affect the mechanical strength in femurs induced by iPTH. These results suggest that NO‐mediated vasorelaxation plays partly a role in the anabolic action of PTH on cortical bone. © 2016 The Authors. Cell Biochemistry and Function published by John Wiley & Sons, Ltd.     

Repeated immunostaining of the same tissue section using alkaline phosphatase as a reporter

One can determine the best dilution of a primary antibody for immunohistochemistry that uses horseradish peroxidase conjugated to a secondary antibody by testing increasing concentrations sequentially on the same tissue section. When the same tissue section is incubated repeatedly with increasing concentrations of primary antibodies to epithelial membrane antigen, smooth muscle α-actin, or vimentin using alkaline phosphatase conjugated to a secondary antibody as the reporter, the best staining was obtained with a less concentrated primary antibody than was optimal for a single staining test. The best concentration of primary antibody for single run staining using an alkaline phosphatase reporting system is usually four times the best concentration for staining with multiple runs. The optimal concentration can be determined by denaturing the residual alkaline phosphatase and extracting residual stain by incubating the section in 4:1 diglyme:phosphate buffered saline for 20 min at 80^o C between tests of primary antibody concentrations. I tested the method for four chromogens from one supplier and one chromogen from a different supplier.     

Investigation of the helicon discharge plasma parameters in a hybrid RF plasma system

Results of an experimental study of the helicon discharge plasma parameters in a prototype of a hybrid RF plasma system equipped with a solenoidal antenna are described. It is shown that an increase in the external magnetic field leads to the formation of a plasma column and a shift of the maximum ion current along the discharge axis toward the bottom flange of the system. The shape of the plasma column can be controlled via varying the configuration of the magnetic field.     

Study of the formation,stability, and X-ray emission of the Z-pinch formed during implosion of fiber arrays at the Angara-5-1 facility

Results from experimental studies on the implosion of arrays made of kapron fibers coated with different metals (Al, In, Sn, and Bi) are presented. It is shown that the power, total energy, and spectrum of radiation emitted by the imploding array depend on the number of metallized fibers and the mass of the metal layer deposited on them but are independent of the metal characteristics (density, atomic number, etc.). Analysis of frame X-ray images shows that the Z-pinches formed in the implosion of metallized kapron fiber arrays are more stable than those formed in wire arrays and that MHD perturbations in them develop at a slower growth rate. Due to the lower rate of plasma production from kapron fibers, the plasma formed at the periphery of the array forms a layer that plays the role of a hohlraum wall partially trapping soft X-ray emission of the Z-pinch formed in the implosion of the material of the deposited metal layer. The closure of the anode aperture doubles the energy of radiation emitted in the radial direction.     

Suppression of 5-lipoxygenase activity by anionic cholesterol derivatives

5-Lipoxygenase (5-LO) is a key enzyme involved into biosynthesis of leukotrienes (LTs), mediating the host defense system, and acting simultaneously as inflammatory agents. In this work the effect of anionic cholesterol derivatives on 5-LO activity has been investigated. Cholesterol sulfate activates human polymorphonuclear leukocytes (PMNL) and stimulates their adhesion to endothelium and collagen. Cholesterol sulfate and cholesterol phosphate suppressed leukotriene production in PMNL and in rat basophil leukemia (RBL-1) cell line as well as in homogenates of these cells. Kinetic characteristics of the effect of anionic cholesterol derivatives on leukotriene synthesis have been obtained. In all experiments cholesterol phosphate (charge-2) was shown to be more potent inhibitor than cholesterol sulfate (charge-1). We believe that this fact highlights the importance of negatively charged ester groups for suppression of 5-LO activity.     

Decay of Electron-Beam-Generated Argon Plasma upon Outflow into Unexcited Gas

Plasma Physics Reports - Decay of plasma excited by injection of a quasi-stationary beam of high-energy electrons into argon filling a cylindrical reaction chamber is studied experimentally. Based...     

Barley microgreen incorporation in diet-controlled diabetes and counteracted aflatoxicosis in rats

Increased environmental pollution and unhealthy lifestyle are blamed for escalated chronic diseases. Exposure to aflatoxins was recently suggested to have a role in the increased incidence of type 2 diabetes mellitus. Diet modification and consumption of different functional food are now gaining attention, especially in diabetes management. This study investigates the effect of a diet containing barley microgreen against diabetes induced by streptozotocin with or without aflatoxin administration in rats. Barley microgreen was rich in 3′-Benzyloxy-5,6,7,4′-tetramethoxyflavone (48.8% of total) followed by 5β,7βH,10α-Eudesm-11-en-1α-ol (18.46%). Streptozotocin injection and/or aflatoxin administration significantly elevated glucose level, decreased insulin level, decreased β-cell function, deteriorated liver and kidney function parameters, and induced oxidative stress in the liver. Histopathology revealed irregular small-sized islets and decreased area % of insulin-positive beta cells in the pancreas, hepatic degeneration, nephropathy, and neuropathy in diabetic and/or aflatoxin administered rats compared to control. Barley microgreen diet fed to diabetic rats with or without aflatoxin alleviated all evaluated parameters. Barley microgreen diet also ameliorated the toxic effect of aflatoxin. In conclusion, exposure to aflatoxin aggravated diabetes and its complication. The incorporation of barley microgreen in the diet was able to control type 2 diabetes mellitus and the improved outcomes observed with barley microgreen treatments involved or occurred in conjunction with improved biomarkers of oxidative stress.     

Disease-associated mutations in the extracytoplasmic loops of cystic fibrosis transmembrane conductance regulator do not impede biosynthetic processing but impair chloride channel stability

Consistent with its function as a chloride channel regulated entirely from the cytoplasmic side of the plasma membrane, the cystic fibrosis transmembrane conductance regulator (CFTR) glycoprotein exposes little of its mass on the exterior surface of cells. The first and fourth extracytoplasmic loops (ELs) contain approximately 15 and 30 residues, respectively; the other four ELs are extremely short. To examine the influence of missense mutants in ELs detected in patients with cystic fibrosis, we have expressed them in mammalian (baby hamster kidney (BHK21)) cells and assessed their biosynthetic processing and chloride channel activity. In contrast to previous findings that 18 of 30 disease-associated missense mutations in cytoplasmic loops caused retention of the nascent polypeptides in the endoplasmic reticulum, all the EL mutants studied matured and were transported to the cell surface. This pronounced asymmetry is consistent with the notion that endoplasmic reticulum quality control of nascent CFTR is exerted primarily on the cytoplasmic side of the membrane. Although this set of EL mutations has little effect on CFTR maturation, most of them seriously compromise its chloride channel activity. Substitutions at six different positions in EL1 and single positions in EL2 and EL4 all destabilized the open state, some of them severely, indicating that the ELs contribute to the stability of the CFTR ion pore.     

Mossbauer spectroscopic study of functional thermal destruction of iron--sulfur proteins in membranes of thermophilic cyanobacteria synechococcus elongatus

A mathematical model of the Mossbauer spectrum (80K) of native membranes of Synechococcus elongatus was constructed on the basis of values of the quadruple splitting (Delta) and the isomeric shift (delta) of the iron-containing components of the photosynthetic apparatus obtained from the literature. Thermally induced changes in the intensity of the spectral components of membranes and isolated preparations of photosystem (PS) I were studied using this model. It was shown that exposure of membranes to 70-80 degrees C causes a decrease in the intensity of the components related to the FX, FA, and FB centers and surface-located ferredoxins of PS I, an increase in the intensity of the doublets of oxidized iron clusters that are nonspecifically absorbed by the membranes, and formation of a new doublet. Spectral parameters of this doublet (Delta = 3.10 mm/sec and delta = 1.40 mm/sec) are typical of inorganic hydrated forms of reduced iron. Heating of PS I preparations also causes a decrease in the intensity of doublets of the FX, FA, and FB centers and an increase in the intensity of doublets of nonspecifically bound oxidized iron. However, this does not cause formation of inorganic reduced iron. Comparison between the intensities of the Mossbauer spectral components in intact and heated samples suggests that the main source of reduced iron in membranes is surface-located ferredoxins. Nonspecifically bound oxidized iron is formed at the expense of the FX, FA, and FB centers. Disappearance of spectral components associated with ferredoxins and accumulation of reduced iron in membranes occur within the temperature range critical for inhibition of electron transport through PS I to oxygen. These findings suggest that the thermally induced processes of accumulation of reduced iron and inhibition of electron transport in PS I in membranes of thermophilic cyanobacteria are interrelated and caused mainly by degradation of the Fe--S centers of ferredoxins. The possible role of reduced iron accumulation in the degradation of the photosynthetic apparatus induced by heat and other extreme physical and chemical factors is discussed.     

Activity of neurons in the anterolateral region of the rabbit motor cortex during taking food and nonfood objects in instrumental behavior

We showed earlier that only 2-4% of N-neurons in the rabbit's anterolateral ("masticatory") motor cortex (AC) that are being specialized in relation to Newly formed acts (such as pressing the pedal, approaching the pedal) during the elaboration of instrumental food-acquistion behavior. The majority of neurons in this area are O-neurons that are specialized in relation to the Older acts formed long before the acquisition of instrumental behavior: mostly taking of food. It was shown also that electrical stimulation of this area produced jaw movements. It might seem reasonable to propose that the number of N-neurons may be increased in this area if pedal pressing in substituted with instrumental act like seizure and pulling a ring that is more corresponding to the "projectional property" of the AC. Here we show that the number of N-neurons in rabbit's AC remains constant -3.4%, even is the animals that were trained to pull a ring instead of pressing a pedal. Thus, we assume that the small number of N-neurons is the stable property of the AC. However, the role of the AC in the subserving of the behavior does not remain the same. Much more AC units (O-neurons) were activated in the ring pulling than in the pedal pressing instrumental act.