Adverse effects of hyperbaric oxygen on [3H]uridine incorporation and uridine kinase activity in B104 rat neuroblastoma cells

The effects of hyperbaric oxygen on uracil nucleotide metabolism in B104 rat neuroblastoma cells were investigated. Cells exposed to 10 atm O₂ for 4 h incorporated markedly less [³H]uridine into the acid-soluble fraction and RNA compared to cells kept in ambient air. The acid-soluble fraction of the oxygen-treated cells contained less total [³H]uridine phosphates ([³H]UMP + [³H]UDP + [³H]UTP) than air-treated cells. Uridine kinase activity, assayed in cytosolic extracts from cells exposed to 10 atm O₂ for 4 h, was decreased by 46% compared to the air controls. The reduced enzyme activity which appears to account for the depressed [³H]uridine incorporation, may contribute to the lethal effects of oxygen in these cells.Abbreviations DMEM Dulbecco's Modified Eagle's Medium - Hepes 4-(2-hydroxyethyl)-1-piperazineethanesulphonic acid     

Increased expression of TNF-alpha,IL-6, and IL-8 in HALS: implications for reduced adiponectin expression and plasma levels

Human immunodeficiency virus (HIV)-associated lipodystrophy syndrome (HALS) is a side effect of highly active antiretroviral therapy of HIV-infected patients; however, the mechanism of the lipodystrophy and insulin resistance seen in this syndrome remains elusive. Adiponectin, an adipocyte-specific protein, is thought to play an important role in regulating insulin sensitivity. We investigated circulating levels and gene expression of adiponectin in subcutaneous abdominal adipose tissue (AT) from 18 HIV-infected patients with HALS compared with 18 HIV-infected patients without HALS. Implications of cytokines for adiponectin levels were investigated by determining circulating levels of TNF-alpha, IL-6, and IL-8 as well as gene expression of these cytokines in AT. HALS patients exhibited 40% reduced plasma adiponectin levels (P < 0.05) compared with non-HALS subjects. Correspondingly, adiponectin mRNA levels in AT were reduced by >50% (P = 0.06). HALS patients were insulin resistant, and a positive correlation was found between plasma adiponectin and insulin sensitivity (r = 0.55, P < 0.01) and percent limb fat (r = 0.61, P < 0.01). AT mRNA of TNF-alpha, IL-6, and IL-8 was increased in AT of HALS subjects (P < 0.05), and both AT TNF-alpha mRNA and plasma TNF-alpha were negatively correlated to plasma adiponectin (P < 0.05). Finally, TNF-alpha was found in vitro to inhibit human AT adiponectin mRNA by 80% (P < 0.05). In conclusion, HALS patients have reduced levels of plasma adiponectin and adiponectin mRNA in AT. Increased cytokine mRNA in AT is hypothesized to exert an inhibitory effect on adiponectin gene expression and, consequently, to play a role in the reduced plasma adiponectin levels found in HALS patients.     

Effect of DHLA on response of isolated rat urinary bladder to repetitive field stimulation

Lipoic acid is an essential coenzyme in the oxidation of pyruvate and -ketoglutarate. It is easily converted to its reduced form, dihydrolipoic acid (DHLA), in vivo thereby forming a redox pair. DHLA is important in the maintenance and integrity of specific neuronal and subcellular membranes. In the present study we investigated the effect of DHLA on the response of isolated rat bladder strips to repetitive field stimulation (FS), a method used to exhaust synaptic stores of acetylcholine resulting in nerve and synaptic damage.Isolated strips of rat urinary bladders were separated into 4 groups. Group 1 strips were incubated with choline + acetyl-CoA; Group 2 strips with choline, acetyl-CoA + DHLA; and Group 3 with DHLA. Group 4 strips were controls. All strips in Groups 1–3 were subjected to 2 h of repetitive FS followed by 2 h of recovery.DHLA had no effect on the progressive decrease in contractile response observed during repetitive stimulation. However, strips incubated in the presence of DHLA showed a significantly greater degree of recovery than strips incubated in the absence of DHLA. We believe that the protection of the contractile response is related to DHLA's ability to protect nerve and/or muscle membranes from oxidative damage.     

Stimulation of the phosphorylation of uridine in skeletal muscle by insulin and vanadate

Summary The action of insulin and sodium vanadate on the phosphorylation of uridine by skeletal muscle was studied in vitro. Insulin significantly increased the incorporation of ³H-uridine into uracil nucleotides by pieces of rat diaphragm incubated for 15 min in a phosphate-buffered medium. This action of the hormone was exceptionally consistent when MgATP was added to the incubation medium. In experiments in which pieces of psoas muscle were incubated in TRIS buffer in the presence and absence of insulin, the hormone caused a significant activation of uridine kinase measured in cytosolic extracts of the incubated tissue. In experiments with rat diaphragm similar to those with insulin, the vanadate ion caused a significant increase in phosphorylation of uridine. The results of these experiments provide preliminary support for the proposal that uracil nucleotide metabolism is regulated by insulin and that insulin activates uridine kinase, the limiting enzyme in the synthesis of uracil nucleotides from uridine by the salvage pathway.     

A comprehensive model of purine uptake by the malaria parasite Plasmodium falciparum: identification of four purine transport activities in intraerythrocytic parasites

Plasmodium falciparum is incapable of de novo purine biosynthesis, and is absolutely dependent on transporters to salvage purines from the environment. Only one low-affinity adenosine transporter has been characterized to date. In the present study we report a comprehensive study of purine nucleobase and nucleoside transport by intraerythrocytic P. falciparum parasites. Isolated trophozoites expressed (i) a high-affinity hypoxanthine transporter with a secondary capacity for purine nucleosides, (ii) a separate high-affinity transporter for adenine, (iii) a low-affinity adenosine transporter, and (iv) a low-affinity/high-capacity adenine carrier. Hypoxanthine was taken up with 12-fold higher efficiency than adenosine. Using a parasite clone with a disrupted PfNT1 (P. falciparum nucleoside transporter 1) gene we found that the high-affinity hypoxanthine/nucleoside transport activity was completely abolished, whereas the low-affinity adenosine transport activity was unchanged. Adenine transport was increased, presumably to partly compensate for the loss of the high-affinity hypoxanthine transporter. We thus propose a model for purine salvage in P. falciparum, based on the highly efficient uptake of hypoxanthine by PfNT1 and a high capacity for purine nucleoside uptake by a lower affinity carrier.     

Being an Only or Last-Born Child Increases Later Risk of Obesity

Background

Studies have suggested that number of siblings and birth order is associated with obesity. However, studies combining these exposures are needed. This study aimed at investigating obesity in children and young adults in regard to different combinations of family size and birth order.

Methods

Two cohorts selected from the general population were investigated: The Copenhagen School Health Records Register (CSHRR) and a Draft Board (DB) sample with measured heights and weights in childhood (age 13 years) and young adulthood (age 19 years), respectively. Information on birth order, number of siblings, and relevant covariates were available on 29 327 children, as well as on 323 obese young men and 575 randomly selected controls of young men representing approximately 58 000. The relation between number of siblings and birth order, respectively, and having a Body Mass Index (BMI) z-score above or equal to the 95^th percentile in childhood or having a BMI of at least 31.00 kg/m² in young adulthood was analysed using logistic regression analyses adjusted for relevant confounders.

Results

Only children had significantly higher odds of obesity both in childhood and in young adulthood compared with children with siblings, odds ratio (OR) = 1.44 (95% Confidence Interval (CI): 1.26–1.66) and OR = 1.76 (95% CI: 1.18–2.61), respectively. No association between first-born status and obesity was found. The OR of last-born children being obese was also significantly increased in childhood, e.g. OR = 1.93 (95% CI: 1.09−3.43) of obesity if last-born in a family of four children. This was not found in young adulthood. Additionally, higher spacing to previous sibling (average 1872 vs. 1303 days; p = 0.026 in four children families) was observed in obese last-born compared to non-obese last-born children.

Conclusion

Being an only or last-born child is associated with obesity. These associations may provide leads to targeted prevention of obesity in children.     

Desaturation of skeletal muscle structural and depot lipids in obese individuals during a very-low-calorie diet intervention

Objective: This study investigated whether a very‐lowcalorie dietary intervention (VLCD) may influence composition of skeletal muscle cell membrane phospholipid and composition and concentration of intramyocellular triglyceride (IMTG) in obese subjects. The working hypothesis proposed that a VLCD would decrease saturated fatty acids (FAs) and increase long‐chain polyunsaturated FAs (LCPUFAs) in muscular structural lipids, as such changes have been associated with improved insulin sensitivity. Research Methods and Procedures: Skeletal muscle biopsies (vastus lateralis) were obtained from 13 obese subjects (nine women) before and after 8 weeks on VLCD (～600 to 800 kcal/d). FA composition in muscle cell membrane phospholipid and concentration and FA composition of IMTG were determined by gas‐liquid chromatography. Results: Baseline BMI was 36.0 ± 3.4 kg/m². Weight loss was 9.3 ± 1.1 kg (8.8 ± 1.1%; p < 0.0001); loss of adipose tissue was 5.9 ± 0.9 kg (p < 0.0001). Insulin resistance (by homeostasis model assessment) decreased (?44 ± 7%; p < 0.001). Muscle cell membrane phospholipid saturated FAs decreased (?3.2 ± 1.3%; p < 0.05), whereas monounsaturated FAs (4.3 ± 1.7%; p < 0.05), LCPUFAs (11 ± 6%; p < 0.05), and the ratio of LCPUFAs to saturated FAs (12 ± 5%; p < 0.05) increased. IMTG decreased, but not significantly (?5%). IMTG‐saturated FAs decreased (?3.3 ± 1.5%; p < 0.05), whereas LCPUFAn‐3 (29 ± 9%; p < 0.01), LCPUFAn‐6 (33 ± 9%; p < 0.01), and the ratio of LCPUFAs to saturated FAs (34 ± 8%; p < 0.001) increased. Plasma total cholesterol (?15 ± 6%; p < 0.05), low‐density lipoprotein‐cholesterol (?16 ± 5%; p < 0.01), high‐density lipoprotein‐cholesterol (?8 ± 2%; p < 0.01), and plasma triglyceride (?19 ± 12%; p = 0.10) all decreased during the VLCD. Discussion: Desaturation of both muscle cell membrane phospholipid and IMTG was significant but modest during a VLCD in obese subjects. Further research must delineate whether such changes in skeletal muscle structural and depot lipid composition themselves are enough to promote the observed improvements in insulin action.