Human iodine requirements determined by the saturation kinetics model

Iodine plays a decisive role in metabolism and the process of early growth and development of most organs, especially of the brain. Effects of iodine deficiency include goiter, stillbirth and miscarriage, neonatal and juvenile thyroid deficiency, dwarfism, mental defects, deaf mutism, spastic weakness and paralysis. In this study, the application of a mathematical model (derived from Machaelis-Menten enzyme kinetics) to iodine measured in urine samples from a randomly selected group derived from the Egyptian village of West El-Mawhoub in the Dakhlah Oasis resulted in the conclusion that iodine excretion parameters can be used to characterize iodine utilization and accurately predict the level of salt iodination required to maintain proper physiological functions. The four parameter saturation kinetics model analysis indicated that a salt iodination level of 63 mg/kg reduced the severity of IDD, with 83% of the studied subjects having urinary excretion levels of 1.18 micromol/L. This gives a convenient mechanism for providing adequate dietary iodine with a non-invasive index for the avoidance of IDD. Commercially available salt was analyzed using standard iodiometric titration methods to determine iodination levels. Analysis revealed that only 20% of the commercially available salt complied with the manufacturer's label and revealed the presence of large individual variability between batches amounting to -95 to +150% of the claimed iodine level. Therefore, salt iodination requires careful supervision to ensure that promised iodine levels are being delivered and consumed.     

Influence of ethylene-oxy spacer group on the activity of linezolid: synthesis of potent antibacterials possessing a thiocarbonyl group

The influence of an ethylene-oxy spacer element between the heterocycle and the aromatic ring in linezolid is reported. The introduction of such spacer group generated compounds with inferior antibacterial activity. However, the conversion of the acetamide group present in the linezolid analogues to either thiocarbamate or thioacetamide functionality restored the activity. The synthesis of linezolid analogues possessing the ethylene-oxy spacer group along with SAR studies with different heterocycles and preparation of some thiocarbonyl compounds possessing potent antibacterial property are presented.     

Oxygen free radical scavenger enzyme polymorphisms in systemic sclerosis

We performed a case-control study of polymorphisms of glutathione S-transferase (GST) isoenzymes and manganese superoxide dismutase (MnSOD) in black South Africans with systemic sclerosis (SSc). The frequency of the GSTM1*B phenotype was significantly decreased in the overall SSc group compared with controls (OR=0.19, p(corr)<.05), implying a possible protective effect against development of the disease. There was also a trend toward increased MnSODAla allele and phenotype frequencies in the diffuse cutaneous SSc subset compared with controls (OR=2.11 and 3.15, respectively, p(corr)<.1). Our findings provide new data on the distribution of GST and MnSOD polymorphisms in healthy Africans and further evidence that genetic factors may have a contributory role to play in predisposing to oxidative stress in SSc.     

Suppression of mitochondria-dependent neutrophil apoptosis with thermal injury

Neutrophil apoptosis is delayed under trauma and/or sepsis conditions. The mechanism for the delay has remained unclear. We hypothesize that modulation of the mitochondrial pathway of apoptosis contributes to the delay in neutrophil apoptosis with burn injury. Rats were subjected to burn injury (30% of total body surface area, 98°C for 10 s) and euthanatized 24 h postinjury. Blood neutrophils from sham and burn-injured rats were isolated by Ficoll gradient centrifugation and cultured for 2 or 8 h. Neutrophil apoptosis was determined by annexin V and propidium iodide (PI) labeling and flow cytometry. Neutrophil mitochondrial morphology was assessed via histochemical staining (MitoTracker GreenFM) and confocal microscopy. Neutrophils from rats with burn injury showed a decreased level of apoptosis compared with sham rat neutrophils at both 2 and 8 h of incubation. In incubated sham rat neutrophils, mitochondria showed a change from normal "tubular" to an "aggregated" morphology. In contrast, cultured neutrophils from burn rats did not exhibit this mitochondrial morphological transition until 8 h of incubation. Compared with sham rat neutrophils, neutrophils from burn rats showed decreased levels of active caspase-9 and -3. Whereas an upregulation of Bcl-xL and a downregulation of Bax seemed to contribute to decreased apoptosis in burn rat neutrophils at 2 h of incubation, the decreased apoptosis at 8 h appeared to be associated with a decrease in Bax and increased phosphorylated Bad. These data suggest that suppression of the mitochondrial pathway plays an essential role in the delay of polymorphonuclear neutrophil apoptosis with burn injury. burn; rat; polymorphonuclear leukocytes; caspase-3; caspase-9; cytochrome c; Bcl-xL; Bax; Bad; MitoTracker GreenFM; confocal microscopy     

Analyses of impact of metal ion contamination on carp (Cyprinus carpio L.) gill cell suspensions

The decline in fish population because of water contamination is problem. As a result of direct exposure in water, it has been readily accepted that the gills are the main site of water contamination and toxicity (e.g., metal ions). In the present study, we investigated metal ion contamination on the functional capacity of carp gill cells with antioxidant interactions in an in vitro study. The extent of cellular membrane damage, lipid peroxidation (LPO) (as TBARS levels), and glutathione (GSH) content were investigated after the addition of two metal ion compounds (viz., CuSO₄ and HgCl₂) in various concentrations (300, 500, 700, 1000, and 3000 μM) to gill cell preparation of the freshwater fish carp (Cyprinus carpio L.) with modulations by bovine serum albumin (BSA) (0.5% and 1.0%) and dimethyl sulfoxide (DMSO) (0.5%) as free-radical scavengers. The Comet assay technique was also performed for the highest concentrations of the two mentioned metal ions as an index of DNA breaks. The outcomes were as follows: (1) Copper and mercury increased the rate of LPO dose dependently (r=+0.995 and r=+0.993, respectively; p<0.001), but the GSH content was only marginally affected (r=−0.787 and r=−0.844, respectively; p<0.05). (2) Depletion of GSH molecules by copper had a wider range than mercury. (3) In the highest concentration of metal ions (3000 μM), both DMSO and 1.0% BSA showed a pro-oxidative potential to elevate the levels of TBARS (p<0.001), but for other concentrations when supplemented with three scavengers, a fall in the levels of the latter was found. (4) The addition of 1.0% BSA to medium containing 3000 μM of metal ions caused a significant decline in GSH content (p<0.01). (5) Copper and mercury could cause a high rate of DNA breaks (single stranded) in carp gill cell suspensions as a Comet appearance. These findings indicate that copper and mercury have a deleterious influence on membrane integrity and GSH content in a relatively dose-dependent manner. The complexes of metal ions and thiol (−SH) residues of cell proteins could also act as a potential cell toxicant leading to disturbances in cell functions causing cell death. DNA fragmentation is frequent in metal ion contamination.     

Biotransformation of hydrocortisone by a natural isolate of Nostoc muscorum

Hydrocortisone was converted in the culture of an isolated strain of the cyanobacterium Nostoc muscorum PTCC 1636 into some androstane and pregnane derivatives. The microorganism was, isolated during a screening program from soil samples collected from paddy fields of north of Iran. The bioproducts obtained were purified using chromatographic methods and identified as 11beta-hydroxytestosterone, 11beta-hydroxyandrost-4-en-3,17-dione and 11beta,17alpha,20beta,21-tetrahydroxypregn-4-en-3-one on the basis of their spectroscopic features.     

Altered expression of the<Emphasis Type="Italic"> Arabidopsis</Emphasis> ortholog of<Emphasis Type="Italic"> DCL</Emphasis> affects normal plant development

The DCL (defective chloroplasts and leaves) gene of tomato (Lycopersicon esculentum Mill.) is required for chloroplast development, palisade cell morphogenesis, and embryogenesis. Previous work suggested that DCL protein is involved in 4.5S rRNA processing. The Arabidopsis thaliana (L.) Heynh. genome contains five sequences encoding for DCL-related proteins. In this paper, we investigate the function of AtDCL protein, which shows the highest amino acid sequence similarity with tomato DCL. AtDCL mRNA was expressed in all tissues examined and a fusion between AtDCL and green fluorescent protein (GFP) was sufficient to target GFP to plastids in vivo, consistent with the localization of AtDCL to chloroplasts. In an effort to clarify the function of AtDCL, transgenic plants with altered expression of this gene were constructed. Deregulation of AtDCL gene expression caused multiple phenotypes such as chlorosis, sterile flowers and abnormal cotyledon development, suggesting that this gene is required in different organs. The processing of the 4.5S rRNA was significantly altered in these transgenic plants, indicating that AtDCL is involved in plastid rRNA maturation. These results suggest that AtDCL is the Arabidopsis ortholog of tomato DCL, and indicate that plastid function is required for normal plant development.Abbreviations DCL Defective chloroplasts and leaves - GFP Green fluorescent protein     

Determination of a glucose-dependent futile recycling rate constant from an intraperitoneal glucose tolerance test

Increased glucose cycling between glucose and glucose-6-phosphate is characteristic of insulin resistance and hyperglycemia seen with Type II diabetes. Traditionally, glucose cycling is determined by the difference between hepatic glucose output measured with separate [2-3H]glucose and [6-3H]glucose infusions. We demonstrate a novel method for determining hepatic glucose recycling from an intraperitoneal glucose tolerance test (IPGTT). A single tracer, [1, 2-13C(2)]glucose (a M2 glucose isotopomer), was administered at 1mg/g body weight to 4-month-old C57BL/6 mice. Hepatic glucose recycling was monitored by the appearance of a plasma M1 isotopomer of glucose, which is produced by the action of the pentose cycle on the M2 glucose isotopomer in the liver. The initial M2 enrichment was 56% and decreased to 13% at the end of 3 h, and the M1 enrichment peaked at 2 h. The ratio of plasma M1/M2 glucose increased linearly with time to approximately 25%, and the regression of the M1/M2 ratio against time gives a slope, termed the in vivo glucose-dependent futile recycling rate constant k(HR). k(HR) estimates glucose/glucose-6-phosphate futile cycling, along with glucose recycling through the pentose cycle. These observations demonstrate complex substrate cycling during an IPGTT using a single stable isotope tracer.     

Interaction of Bordetella pertussis adenylate cyclase with CD11b/CD18: Role of toxin acylation and identification of the main integrin interaction domain

Adenylate cyclase toxin (CyaA) is one of the major virulence factors produced by Bordetella pertussis, the whooping cough agent. CyaA belongs to the repeat in toxin protein family and requires a post-translational fatty acylation to form cation-selective channels in target cell membranes and to penetrate into cytosol. We have demonstrated recently that CyaA uses the alphaMbeta2 integrin (CD11b/CD18) as a specific cellular receptor. Here we show that the acylation of CyaA is required for a productive and tight interaction of the toxin with cells expressing CD11b. In addition, we demonstrate that the catalytic domain is not required for binding of CyaA to CD11b and that the main integrin interacting domain of CyaA is located in its glycine/aspartate-rich repeat region. These data decipher, for the first time, the interaction of CyaA with CD11b-positive cells and open new prospects for understanding the interaction of Bordetella pertussis with innate and adaptive immune systems.