Amino acid sequence homologies between rabbit, rat, and human serum retinol-binding proteins

The main transporting protein for vitamin A in rabbit serum, the retinol-binding protein (RBP), was isolated and its amino acid sequence determined. Rabbit RBP was found to be highly homologous to human RBP, whose amino acid sequence was elucidated earlier, and to rat RBP. The rat RBP sequence was obtained by combining information deduced from the nucleotide sequences of two overlapping cDNA clones with the NH2-terminal sequence of the isolated protein determined by automated Edman degradation. The identity between the three proteins is approximately 90%. The high degree of homology between RBP molecules from different species is probably explained by the fact that RBP participates in at least three types of molecular interactions: in the binding of prealbumin, in the interaction with retinol, and in the recognition of a specific cell surface receptor. All these interactions should lead to a conservation of RBP structure. The amino acid differences between rabbit, rat, and human RBP are discussed in light of the recent elucidation of the three-dimensional structure of human RBP. Hybridization of a probe isolated from a rat RBP cDNA clone to restriction enzyme-digested genomic DNA from rat and mouse suggests that RBP is encoded by a single gene.     

A moderate-intensity exercise program fulfilling the American College of Sports Medicine net energy expenditure recommendation improves health outcomes in premenopausal women

The purpose of this study was to assess and quantify the health outcomes associated with a moderate-intensity (50% VO2R) exercise program designed to achieve the American College of Sports Medicine net caloric expenditure guideline of 1,000 kcal x wk(-1). Fifteen apparently healthy but sedentary premenopausal women with the baseline characteristics (mean +/- SD age, height, weight, body composition, and VO2max: 37.4 +/- 6.3 yr, 166.2 +/- 6.2 cm, 72.1 +/- 11.2 kg, 32.5 +/- 5.8%, and 34.8 +/- 5.8 mL x kg(-1) x min(-1), respectively) participated in and completed the study. Exercise training was performed 3-4 days per week for 10 weeks in a progressive manner at moderate intensity (50% VO2R). There were significant (P < 0.05) improvements in VO2max (+2.5 mL x kg(-1) x min(-1)), systolic (-13.7 mm Hg) and diastolic (-6.4 mm Hg) blood pressure, high-density lipoprotein cholesterol (+3.2 mg x dL(-1)), fasting blood glucose (-4.9 mg x dL(-1)), and percent body fat (-1.6%). Although the American College of Sports Medicine specifies that the energy expenditure goal should be a net caloric expenditure of 1,000 kcal x wk(-1) and classifies relative moderate intensity as 40-59% of heart rate reserve or VO2R, we are unaware of any previous investigations that have examined the specific health outcomes associated with an exercise program fulfilling these requirements. Results indicate that significant health benefits will be conferred to previously sedentary, premenopausal women who engage in a moderate-intensity, 10-week exercise program designed to fulfill the net energy expenditure guideline of 1,000 kcal x wk(-1).     

The influence of pH on the specific adhesion of P piliated Escherichia coli

Adhesion to host tissues is an initiating step in a majority of bacterial infections. In the case of Gram-negative bacteria this adhesion is often mediated by a specific interaction between an adhesin, positioned at the distal end of bacterial pili, and its receptor on the surface of the host tissue. Furthermore, the rod of the pilus, and particularly its biomechanical properties, is believed to be crucial for the ability of bacteria to withstand external forces caused by, for example, (in the case of urinary tract infections) urinary rinsing flows by redistributing the force to several pili. In this work, the adhesion properties of P-piliated E. coli and their dependence of pH have been investigated in a broad pH range by both the surface plasmon resonance technique and force measuring optical tweezers. We demonstrate that P piliated bacteria have an adhesion ability throughout the entire physiologically relevant pH range (pH 4.5 - 8). We also show that pH has a higher impact on the binding rate than on the binding stability or the biomechanical properties of pili; the binding rate was found to have a maximum around pH 5 while the binding stability was found to have a broader distribution over pH and be significant over the entire physiologically relevant pH range. Force measurements on a single organelle level show that the biomechanical properties of P pili are not significantly affected by pH.     

Changes in EMG activity in the upper trapezius muscle due to local vibration exposure

Exposure to vibration is suggested as a risk factor for developing neck and shoulder disorders in working life. Mechanical vibration applied to a muscle belly or a tendon can elicit a reflex muscle contraction, also called tonic vibration reflex, but the mechanisms behind how vibration could cause musculoskeletal disorders has not yet been described. One suggestion has been that the vibration causes muscular fatigue. This study investigates whether vibration exposure changes the development of muscular fatigue in the trapezius muscle. Thirty-seven volunteers (men and women) performed a sub-maximal isometric shoulder elevation for 3 min. This was repeated four times, two times with induced vibration and two times without. Muscle activity was measured before and after each 3-min period to look at changes in the electromyography parameters. The result showed a significantly smaller mean frequency decrease when performing the shoulder elevation with vibration (?2.51 Hz) compared to without vibration (?4.04 Hz). There was also a slightly higher increase in the root mean square when exposed to vibration (5.7% of maximal voluntary contraction) compared to without (3.8% of maximal voluntary contraction); however, this was not statistically significant. The results of the present study indicate that short-time exposure to vibration has no negative acute effects on the fatiguing of upper trapezius muscle.     

Actomyosin motility on nanostructured surfaces

We have here, for the first time, used nanofabrication techniques to reproduce aspects of the ordered actomyosin arrangement in a muscle cell. The adsorption of functional heavy meromyosin (HMM) to five different resist polymers was first assessed. One group of resists (MRL-6000.1XP and ZEP-520) consistently exhibited high quality motility of actin filaments after incubation with HMM. A second group (PMMA-200, PMMA-950, and MRI-9030) generally gave low quality of motility with only few smoothly moving filaments. Based on these findings electron beam lithography was applied to a bi-layer resist system with PMMA-950 on top of MRL-6000.1XP. Grooves (100-200nm wide) in the PMMA layer were created to expose the MRL-6000.1XP surface for adsorption of HMM and guidance of actin filament motility. This guidance was quite efficient allowing no U-turns of the filaments and approximately 20 times higher density of moving filaments in the grooves than on the surrounding PMMA.     

Saprobic sequence within the genus euglena

In total 73 species of Euglena were ordered according to their saprobic index (S) and saprobic valency starting with species tolerating a heavy organic pollution and ending with those from most clean water. This paper is meant as an addition to the Ergebnisse der Limnologie, Heft 7 (Sládeek, 1973).     

Recovery of proteins on a milligram scale from polyacrylamide electrophoresis gels,exemplified by purification of a retinol-binding protein

An apparatus suitable for the recovery of proteins from polyacrylamide gels on a milligram scale by displacement electrophoresis (isotachophoresis) is described along with a buffer system that is suitable for this purpose with most proteins. The technique is illustrated by the recovery of a protein from a 15% polyacrylamide gel. The recovery was almost quantitative and the eluted protein showed little contamination upon quantitative amino acid analysis and automatic Edman degradation.     

Helicobacter pylori Cholesteryl α-Glucosides Contribute to Its Pathogenicity and Immune Response by Natural Killer T Cells

Approximately 10–15% of individuals infected with Helicobacter pylori will develop ulcer disease (gastric or duodenal ulcer), while most people infected with H. pylori will be asymptomatic. The majority of infected individuals remain asymptomatic partly due to the inhibition of synthesis of cholesteryl α-glucosides in H. pylori cell wall by α1,4-GlcNAc-capped mucin O-glycans, which are expressed in the deeper portion of gastric mucosa. However, it has not been determined how cholesteryl α-glucosyltransferase (αCgT), which forms cholesteryl α-glucosides, functions in the pathogenesis of H. pylori infection. Here, we show that the activity of αCgT from H. pylori clinical isolates is highly correlated with the degree of gastric atrophy. We investigated the role of cholesteryl α-glucosides in various aspects of the immune response. Phagocytosis and activation of dendritic cells were observed at similar degrees in the presence of wild-type H. pylori or variants harboring mutant forms of αCgT showing a range of enzymatic activity. However, cholesteryl α-glucosides were recognized by invariant natural killer T (iNKT) cells, eliciting an immune response in vitro and in vivo. Following inoculation of H. pylori harboring highly active αCgT into iNKT cell-deficient (Jα18^−/−) or wild-type mice, bacterial recovery significantly increased in Jα18^−/− compared to wild-type mice. Moreover, cytokine production characteristic of Th1 and Th2 cells dramatically decreased in Jα18^−/− compared to wild-type mice. These findings demonstrate that cholesteryl α-glucosides play critical roles in H. pylori-mediated gastric inflammation and precancerous atrophic gastritis.     

Increased Expression of the Very Low-Density Lipoprotein Receptor Mediates Lipid Accumulation in Clear-Cell Renal Cell Carcinoma

Clear-cell renal cell carcinoma (RCC) is, in most cases, caused by loss of function of the tumor suppressor gene von Hippel–Lindau, resulting in constitutive activation of hypoxia-inducible factor (HIF)-1α and expression of hypoxia-induced genes in normoxic conditions. Clear-cell RCC cells are characterized histologically by accumulation of cholesterol, mainly in its ester form. The origin of the increased cholesterol remains unclear, but it is likely explained by an HIF-1α-driven imbalance between cholesterol uptake and excretion. Here, we showed that expression of the very low-density lipoprotein receptor (VLDL-R) was significantly increased in clear-cell RCC human biopsies compared with normal kidney tissue. Partial knockdown of HIF-1α in clear-cell RCC cells significantly reduced the VLDL-R expression, and knockdown of either HIF-1α or VLDL-R reduced the increased lipid accumulation observed in these cells. We also showed increased uptake of fluorescently labeled lipoproteins in clear-cell RCC cells, which was significantly reduced by knockdown of HIF-1α or VLDL-R. Taken together, our results support the concept that the pathological increase of HIF-1α in clear-cell RCC cells upregulates VLDL-R, which mediates increased uptake and accumulation of lipids. These results explain the morphological characteristics of clear-cell RCC, and open up novel possibilities for detection and treatment of clear-cell RCC.