Inhibition of ileal brush-border chloride conductance by specific antibody

Summary Antibody raised in mice was used in attempting to identify proteins responsible for the conductive chloride transport that can be measured in porcine ileal brush border membrane vesicles. Ileal brush-border membrane vesicle protein from pig was separated into five different molecular mass fractions by preparative SDS polyacrylamide disc gel electrophoresis. Separated protein fractions were used to immunize mice. Antibody was screened for reactivity with antigen by Western blotting, and for effects on conductive chloride transport in ileal brush border membrane vesicles. Immunization with brush-border protein from fraction I proteins (>110 kDa) produced polyclonal antisera which specifically inhibited the conductive component of chloride uptake by ileal brush border vesicle preparations. Western blotting of the antigen showed the presence of several protein species of molecular mass >100 kDa that were recognized by immune serum. Spleen cells from a mouse producing antiserum that inhibited conductive chloride transport were fused with a myeloma cell line. The resulting hybridoma colonies produced antibody that reacted with at least seven distinct protein bands by Western blot assay and inhibited chloride conductance in brush-border membrane vesicles.     

Repeated oral administration of capsaicin increases anxiety-like behaviours with prolonged stress-response in rats

This study was conducted to examine the psycho-emotional effects of repeated oral exposure to capsaicin, the principal active component of chili peppers. Each rat received 1 mL of 0.02% capsaicin into its oral cavity daily, and was subjected to behavioural tests following 10 daily administrations of capsaicin. Stereotypy counts and rostral grooming were significantly increased, and caudal grooming decreased, in capsaicin-treated rats during the ambulatory activity test. In elevated plus maze test, not only the time spent in open arms but also the percent arm entry into open arms was reduced in capsaicin-treated rats compared with control rats. In forced swim test, although swimming duration was decreased, struggling increased in the capsaicin group, immobility duration did not differ between the groups. Repeated oral capsaicin did not affect the basal levels of plasma corticosterone; however, the stress-induced elevation of plasma corticosterone was prolonged in capsaicin treated rats. Oral capsaicin exposure significantly increased c-Fos expression not only in the nucleus tractus of solitarius but also in the paraventricular nucleus. Results suggest that repeated oral exposure to capsaicin increases anxiety-like behaviours in rats, and dysfunction of the hypothalamic-pituitary-adrenal axis may play a role in its pathophysiology.     

Dietary nitrate's effects on exercise performance in heart failure with reduced ejection fraction (HFrEF)

Heart failure with reduced ejection fraction (HFrEF) is a deadly and disabling disease. A key derangement contributing to impaired exercise performance in HFrEF is decreased nitric oxide (NO) bioavailability. Scientists recently discovered the inorganic nitrate pathway for increasing NO. This has advantages over organic nitrates and NO synthase production of NO. Small studies using beetroot juice as a source of inorganic nitrate demonstrate its power to improve exercise performance in HFrEF. A larger-scale trial is now underway to determine if inorganic nitrate may be a new arrow for physicians' quiver of HFrEF treatments.     

三氧化二砷对食管癌细胞增殖和热休克蛋白70表达的影响

目的：研究三氧化二砷（As2O3）对食管癌细胞增殖和热休克蛋白70（HSP70）表达的影响。方法：通过相差显微镜、流式细胞术、免疫细胞化学染色和免疫印迹分析等方法观察As2O3对人食管癌细胞株EC1的作用效果和作用机制。结果：与对照组相比,经2μmol/L和5μmol/Las2O3作用的细胞出现明显的生长抑制,G2/M期细胞比例增加;2μmol/Las2O3作用48h后经Ecl细胞HSP70（heat shock protein70）及HSC70(heat shock cognate protein70)表达均增加。结论：As2O3诱导食管癌细胞G2/M期阻滞抑制细胞增殖和生长;HSP70的升高是细胞对As2O3作用后出现的应激反应,并与细胞周期阻滞相关。     

Effect of weight loss and exercise on angiogenic factors in the circulation and in adipose tissue in obese subjects

Background:

Vascular growth is a prerequisite for adipose tissue (AT) development and expansion. Some AT cytokines and hormones have effects on vascular development, like vascular endothelial growth factor (VEGF‐A), angiopoietin (ANG‐1), ANG‐2 and angiopoietin‐like protein‐4 (ANGPTL‐4).

Methods:

In this study, the independent and combined effects of diet‐induced weight loss and exercise on AT gene expression and proteins levels of those angiogenic factors were investigated. Seventy‐nine obese males and females were randomized to: 1. Exercise‐only (EXO; 12‐weeks exercise without diet‐restriction), 2. Hypocaloric diet (DIO; 8‐weeks very low energy diet (VLED) + 4‐weeks weight maintenance diet) and 3. Hypocaloric diet and exercise (DEX; 8‐weeks VLED + 4‐weeks weight maintenance diet combined with exercise throughout the 12 weeks). Blood samples and fat biopsies were taken before and after the intervention.

Results:

Weight loss was 3.5 kg in the EXO group and 12.3 kg in the DIO and DEX groups. VEGF‐A protein was non‐significantly reduced in the weight loss groups. ANG‐1 protein levels were significantly reduced 22‐25% after all three interventions (P < 0.01). The ANG‐1/ANG‐2 ratio was also decreased in all three groups (P < 0.05) by 27‐38%. ANGPTL‐4 was increased in the EXO group (15%, P < 0.05) and 9% (P < 0.05) in the DIO group. VEGF‐A, ANG‐1, and ANGPTL‐4 were all expressed in human AT, but only ANGPTL‐4 was influenced by the interventions.

Conclusions:

Our data show that serum VEGF‐A, ANG‐1, ANG‐2, and ANGPTL‐4 levels are influenced by weight changes, indicating the involvement of these factors in the obese state. Moreover, it was found that weight loss generally was associated with a reduced angiogenic activity in the circulation.     

Racial differences in lipid metabolism in women with abdominal obesity

We evaluated palmitate rate of appearance (R(a)) in plasma during basal conditions and during a four-stage epinephrine infusion plus pancreatic hormonal clamp in nine white and nine black women with abdominal obesity, who were matched on fat-free mass, total and percent body fat, and waist-to-hip circumference ratio. On the basis of single-slice magnetic resonance imaging analysis, black women had the same amount of subcutaneous abdominal fat but less intra-abdominal fat than white women (68 +/- 9 vs. 170 +/- 14 cm(2), P < 0.05). Basal palmitate R(a) was lower in black than in white women (1.95 +/- 0.26 vs. 2.88 +/- 0.23 micromol. kg fat-free mass(-1). min(-1), P < 0.005), even though plasma insulin and catecholamine concentrations were the same in both groups. Palmitate R(a) across a physiological range of plasma epinephrine concentrations remained lower in black women, because the increase in palmitate R(a) during epinephrine infusion was the same in both groups. We conclude that basal and epinephrine-stimulated palmitate R(a) is lower in black than in white women with abdominal obesity. The differences in basal palmitate kinetics are not caused by alterations in plasma insulin or catecholamine concentrations or lipolytic sensitivity to epinephrine. The lower rate of whole body fatty acid flux and smaller intra-abdominal fat mass may have clinical benefits because of the relationship between excessive fatty acid availability and metabolic diseases.     

Influence of weekend lifestyle patterns on body weight

Objective: To determine whether alterations in diet and/or activity patterns during weekends contribute to weight gain or hinder weight loss. Methods and Procedures: Randomized, controlled trial comparing 1 year of caloric restriction (CR) with 1 year of daily exercise (EX). Subjects included 48 healthy adults (30F, 18M) aged 50–60 years with BMI 23.5–29.9 kg/m². Body weight was measured on 7 consecutive mornings for a total of 165 weeks at baseline and 437 weeks during the 1‐year interventions. Daily weight changes were calculated for weekends (Friday to Monday) and weekdays (Monday to Friday). Daily energy intake was estimated using food diaries; daily physical activity was measured using accelerometers. Both measures were validated against doubly labeled water (DLW). Results: At baseline, participants consistently gained weight on weekend days (+0.06 ± 0.03 kg/day, (mean ± s.e.), P = 0.02), but not on weekdays (?0.02 ± 0.02 kg/day, P = 0.18). This was attributable to higher dietary intake on Saturdays and lower physical activity on Sundays relative to weekdays (both P < 0.05). During the interventions, both CR and EX participants were in negative energy balance on weekdays (P < 0.005). On weekends, however, CR participants stopped losing weight, and EX participants gained weight (+0.08 ± 0.03 kg/day, P < 0.0001) due to higher dietary intakes on weekends. This helps to explain the slower‐than‐expected rate of weight loss during the interventions. Discussion: Alterations in lifestyle behaviors on weekends contribute to weight gain or cessation of weight loss on weekends. These results provide one explanation for the relatively slow rates of weight loss observed in many studies, and the difficulty with maintaining significant weight loss.     

Effect of chronic treatment with Rosiglitazone on Leydig cell steroidogenesis in rats: <Emphasis Type="Italic">In vivo</Emphasis> and <Emphasis Type="Italic">ex vivo</Emphasis> studies

Background  

The present study was designed to examine the effect of chronic treatment with rosiglitazone - thiazolidinedione used in the treatment of type 2 diabetes mellitus for its insulin sensitizing effects - on the Leydig cell steroidogenic capacity and expression of the steroidogenic acute regulatory protein (StAR) and cholesterol side-chain cleavage enzyme (P450scc) in normal adult rats.     

Particle films for managing arthropod pests of apple

A two-season study showed that a calendar-based spray program to manage arthropod pests with kaolin (60 g/liter) applied at the rate of 450 liters/ha was effective against European apple sawfly, Hoplocampa testudinea (Klug) (Hymenoptera: Tenthredinidae); white apple leafhopper, Typhlocyba pomaria McAtee (Homoptera: Cicadellidae); apple red bug, Lygidea mendax Reuter (Heteroptera: Miridae); pear plant bug, Lygocoris communis (Knight) (Heteroptera: Miridae); and the apple rust mite, Aculus schlechtendali (Nalepa) (Acari: Eriophyidae). Although it reduced Curculionidae damage, the level of damage was still too high. It had no effect on apple maggot, Rhagoletis pomonella (Walsh) (Diptera: Tephritidae); codling moth, Cydia pomonella (L.) (Lepidoptera: Tortricidae); and tarnished plant bug, Lygus lineolaris (Palisot de Beauvois) (Heteroptera: Miridae). Laboratory studies showed fewer gravid twospotted spider mites, Tetranychus urticae Koch (Acari: Tetranychidae), and fewer eggs laid by these females. The study also showed no effect of kaolin on Neoseiulus fallacis (Garman) (Acari: Phytoseiidae).     

Calorie restriction reduces biomarkers of cellular senescence in humans

Calorie restriction (CR) with adequate nutrient intake is a potential geroprotective intervention. To advance this concept in humans, we tested the hypothesis that moderate CR in healthy young-to-middle-aged individuals would reduce circulating biomarkers of cellular senescence, a fundamental mechanism of aging and aging-related conditions. Using plasma specimens from the Comprehensive Assessment of Long-term Effects of Reducing Intake of Energy (CALERIE™) phase 2 study, we found that CR significantly reduced the concentrations of several senescence biomarkers at 12 and 24 months compared to an ad libitum diet. Using machine learning, changes in biomarker concentrations emerged as important predictors of the change in HOMA-IR and insulin sensitivity index at 12 and 24 months, and the change in resting metabolic rate residual at 12 months. Finally, using adipose tissue RNA-sequencing data from a subset of participants, we observed a significant reduction in a senescence-focused gene set in response to CR at both 12 and 24 months compared to baseline. Our results advance the understanding of the effects of CR in humans and further support a link between cellular senescence and metabolic health.