Kinetic and structural analysis of a bacterial protein tyrosine phosphatase-like myo-inositol polyphosphatase

PhyA from Selenomonas ruminantium (PhyAsr), is a bacterial protein tyrosine phosphatase (PTP)-like inositol polyphosphate phosphatase (IPPase) that is distantly related to known PTPs. PhyAsr has a second substrate binding site referred to as a standby site and the P-loop (HCX5R) has been observed in both open (inactive) and closed (active) conformations. Site-directed mutagenesis and kinetic and structural studies indicate PhyAsr follows a classical PTP mechanism of hydrolysis and has a broad specificity toward polyphosphorylated myo-inositol substrates, including phosphoinositides. Kinetic and molecular docking experiments demonstrate PhyAsr preferentially cleaves the 3-phosphate position of Ins P6 and will produce Ins(2)P via a highly ordered series of sequential dephosphorylations: D-Ins(1,2,4,5,6)P5, Ins(2,4,5,6)P4, D-Ins(2,4,5)P3, and D-Ins(2,4)P2. The data support a distributive enzyme mechanism and suggest the PhyAsr standby site is involved in the recruitment of substrate. Structural studies at physiological pH and high salt concentrations demonstrate the "closed" or active P-loop conformation can be induced in the absence of substrate. These results suggest PhyAsr should be reclassified as a D-3 myo-inositol hexakisphosphate phosphohydrolase and suggest the PhyAsr reaction mechanism is more similar to that of PTPs than previously suspected.     

Expression of collagen type I,II, X and Ki-67 in osteochondroma compared to human growth plate cartilage

In order to characterize the consequences for the process of endochondral ossification we performed an immunohistochemical study and compared the expression of collagen type I, II and X as markers of cartilage differentiation and Ki-67 as a marker of cell proliferation in solitary (7-26 years, n=9) and multiple (11-42 years, n=6) osteochondromas with their expression in human fetal and postnatal growth plates. In fetal and young postnatal controls, we found a thin superficial layer of articular cartilage that stained positive for collagen type I while collagen II was expressed in the rest of the cartilage and collagen type X was restricted to the hypertrophic zone. Osteochondromas from children showed lobular collagen type II-positive areas surrounded by collagen type I. In adults, the separation of collagen type I- and type II-positive areas was more blurred, or the cartilaginous cap was missing. Collagen type X was detected in a pericellular distribution pattern within hypertrophic zones but also deeper between bone trabecula. The proliferative activity of osteochondromas from children younger than 14 years of age was comparable to postnatal growth plates, whereas in cartilage from individuals older than 14 years of age, we could not detect significant proliferative activity.     

The impact of weight stigma on caloric consumption

The present study assessed the influence of exposure to weight stigma on energy intake in both overweight and normal-weight adult women. Seventy-three women (mean age: 31.71 ± 12.72 years), both overweight (n = 34) and normal weight (n = 39), were randomly assigned to view one of two videos depicting either weight stigmatizing material or neutral material, after which they consumed snacks ad libitum. Pre- and post-video measures included blood pressure, attitudes toward overweight individuals, and positive and negative affect. Participants' body weight was measured, as was the number of kilocalories consumed following video exposure. Overweight women who watched the stigmatizing video ate more than three times as many kilocalories as overweight women who watched the neutral video (302.82 vs. 89.00 kcal), and significantly more calories than the normal-weight individuals who watched either the stigmatizing or the neutral video. A two-by-two analysis of covariance revealed that even after adjusting for relevant covariates, there was a significant interaction between video type and weight status in that when overweight, individuals consumed significantly more calories if they were in the stigmatizing condition vs. the neutral condition (F(1,65) = 4.37, P = 0.04, η(2) = 0.03). These findings suggest that among overweight women, exposure to weight stigmatizing material may lead to increased caloric consumption. This directly challenges the notion that pressure to lose weight in the form of weight stigma will have a positive, motivating effect on overweight individuals.     

Public Opinion About Laws to Prohibit Weight Discrimination in the United States

Weight discrimination is pervasive in American society and impairs quality of life for obese persons. With approximately two‐thirds of Americans now overweight or obese, vast numbers of people are vulnerable to weight prejudice and its consequences. Currently, no laws exist to prohibit weight discrimination. This study conducted an online survey with a national sample of 1,001 adults (representing demographics of the United States) to examine public support for six potential legislative measures to prohibit weight discrimination in the United States. Results indicated substantial support (65% of men, 81% of women) for laws to prohibit weight discrimination in the workplace, especially for legal measures that would prohibit employers from refusing to hire, terminate, or deny promotion based on a person's body weight. Laws that proposed extending the same protections to obese persons as people with physical disabilities received the least support, suggesting that Americans may not be in favor of considering obesity as a disability. Findings also highlight specific predictors of support (related to sex, age, education, income, body weight, and political ideology). These findings can be used to inform policy makers in efforts to develop antidiscrimination laws. Such measures will rectify health disparities for overweight Americans and facilitate public health efforts to address obesity.     

Additive genetic variance and covariance between relatives in synthetic wheat crosses with variable parental ploidy levels

Cultivated bread wheat (Triticum aestivum L.) is an allohexaploid species resulting from the natural hybridization and chromosome doubling of allotetraploid durum wheat (T. turgidum) and a diploid goatgrass Aegilops tauschii Coss (Ae. tauschii). Synthetic hexaploid wheat (SHW) was developed through the interspecific hybridization of Ae. tauschii and T. turgidum, and then crossed to T. aestivum to produce synthetic hexaploid wheat derivatives (SHWDs). Owing to this founding variability, one may infer that the genetic variances of native wild populations vs improved wheat may vary due to their differential origin and evolutionary history. In this study, we partitioned the additive variance of SHW and SHWD with respect to their breed origin by fitting a hierarchical Bayesian model with heterogeneous covariance structure for breeding values to estimate variance components for each breed category, and segregation variance. Two data sets were used to test the proposed hierarchical Bayesian model, one from a multi-year multi-location field trial of SHWD and the other comprising the two species of SHW. For the SHWD, the Bayesian estimates of additive variances of grain yield from each breed category were similar for T. turgidum and Ae. tauschii, but smaller for T. aestivum. Segregation variances between Ae. tauschii—T. aestivum and T. turgidum—T. aestivum populations explained a sizable proportion of the phenotypic variance. Bayesian additive variance components and the Best Linear Unbiased Predictors (BLUPs) estimated by two well-known software programs were similar for multi-breed origin and for the sum of the breeding values by origin for both data sets. Our results support the suitability of models with heterogeneous additive genetic variances to predict breeding values in wheat crosses with variable ploidy levels.     

Impaired resistance to oxidation of low density lipoprotein in cystic fibrosis: Improvement during vitamin E supplementation

Antioxidants such as vitamin E protect unsaturated fatty acids of LDL against oxidation. In the ex vivo model used, LDL was exposed to Cu²⁺ ions, a potent prooxidant capable of initiating the oxidation of LDL. The lag time, indicating the delay of conjugated diene formation in LDL due to antioxidant protection, was measured in 54 cystic fibrosis (CF) patients with plasma -tocopherol levels below (Group A, n = 30) or above (Group B, n = 24) 15.9 μmol/L (mean - 2 SD of Swiss population). Patients were reevaluated after 2 months on 400 IU/d of oral RRR--tocopherol. In group A, -tocopherol concentrations in LDL increased significantly from 3.2 ± 1.6 mol/mol LDL to 8.2 ± 2.8 mol/mol (P < 0.001) and lag times increased from 79 ± 33, min to 126 ± 48 min (P < 0.001), whereas in the vitamin E sufficient group B no further increase neither in LDL -tocopherol concentrations or in lag times was observed. LDL oleic acid concentrations were higher, and linoleic acid concentrations were lower in patients than in controls. After efficient vitamin E supplementation, lag times were positively related to LDL -tocopherol (P < 0.01) and negatively to LDL linoleic and arachidonic acid content (P < 0.001). The maximum rate of oxidation correlated positively with linoleic and arachidonic acid concentrations, as did the maximum conjugated diene absorbance. These results indicate that LDL resistance to oxidation is impaired in vitamin E deficient CF patients but can be normalized within 2 months when -tocopherol is given in sufficient amounts. Linoleic and arachidonic acid content exhibit a major influence on the LDL resistance to oxidation.     

Ethnic differences in anthropometric characteristics of young children and their parents

The distribution of body fat, or fat patterning, is an important risk factor for cardiovascular disease and diabetes, independent of obesity. Furthermore, the incidence of cardiovascular disease and diabetes varies by ethnicity. We documented ethnic differences in anthropometric characteristics and body fat distribution between Anglo, Black, and Mexican American men (n = 101), women (n = 245), boys (n = 111), and girls (n = 111). We used aggregates of skinfold measures to examine ethnic differences in the deposition of fat in body compartments (body, trunk, leg, and arm) and analyzed trunk-extremity skinfold ratios to determine which best reflected ethnic differences in fat distribution. The results show that Mexican American mothers have larger skinfold ratios and more body fat (as determined by skinfold aggregates) than either Anglo or Black American mothers, whereas Black American mothers have larger ratios than Anglo American mothers. Mexican American fathers also have larger skinfold ratios but not more body fat (skinfold aggregates) than Anglo American fathers. Mexican American fathers have more body fat than Black American fathers, but we found no differences between skinfold ratios. The ethnic differences among children in skinfold ratios and aggregates are similar to those found among fathers, with more differences among girls than boys. Fat patterning differences do exist among the three ethnicities, with greater trunk fat among Mexican and Black Americans. Those ethnicities are known to be at higher risk for cardiovascular disease and diabetes.     

STIMULATION OF NITRATE NET UPTAKE AND REDUCTION BY RED AND BLUE LIGHT AND REVERSION BY FAR-RED LIGHT IN THE GREEN ALGA ULVA RIGIDA1

The steady-state levels of nitrate, nitrite, and ammonium were estimated in the green alga Ulva rigida C. Agardh in darkness after addition of 0.5 mM KNO₃ and irradiation with red (R) and blue (B) light pulses of different duration (5 and 30 min). The net uptake of nitrate was very rapid. Seventy-five percent of the nitrate added was consumed after 60 min in darkness. Although uptake was stable after R or B, efflux of nitrate occurred within 3 h in the dark control and when R or B were followed by far-red (FR) irradiation. The internal nitrate concentration after 3 h in darkness was similar after R and B light pulses; however, the intracellular ammonium was higher after R than after B. The intracellular nitrate and ammonium decreased when FR tight pulses were applied immediately after R or B. Thus, the involvement of phytochrome in the transport of nitrate and ammonium is proposed. Nitrate reductase activity, measured by the in situ method, was increased by both R and B light pulses. The effect was partially reversed by FR light. Nitrate reductase activity was higher after 5 min of R light than after 5 min of B. However, after 30-min light pulses, the relative increase in activity was reversed for R and B. We propose that phytochrome and a blue-light photoreceptor are involved in regulation of nitrogen metabolism. Nitrate uptake and reduction correlates with previously detected light-regulated accumulation of protein in Ulva rigida under the same experimental conditions.