Detection of combined genomic variants in a Jordanian family with familial non-autoimmune hyperthyroidism

Five patients, four brothers and their paternal aunt, presented with a history of overt hyperthyroidism and goiter. Hyperthyroidism in this family was remarkable for its poor response to carbimazole (30–50 mg/d). The thyroid ultrasound showed a diffusely enlarged gland in all the affected members, and thyroid stimulating antibodies (TSAB) were negative. Screening for germline mutations in thyroid stimulating hormone (TSH) receptor (TSHR) gene was performed by direct sequencing of genomic DNA extracted from peripheral blood leukocytes of all family members. The sequence analysis of all TSHR gene exons and intron borders revealed two genomic variants. The first was a single nucleotide polymorphism (SNP) within exon seven (Asn187Asn), whereas the other was located in intron seven (IVS7+68T>G). All affected members, two asymptomatic brothers with sub-clinical hyperthyroidism, and their father were heterozygous for those two genomic variants. Anti-thyroid drug treatment for several months successfully relieved symptoms in one subject, whereas the remaining patients required total thyroidectomy to control their disease. This is the first Jordanian family with familial non-autoimmune hyperthyroidism, with mutations affecting the TSHR gene.     

Nobiletin protects against diabetes-induced testicular injury via hypophysis–gonadal axis upregulation and amelioration of oxidative stress

Background

Testicular injury is one of the most serious problems associated with diabetes mellitus. The present study aimed to compare the effects of two different doses of nobiletin and analyze its mechanisms of action against diabetes-induced testicular impairment in rats.

Methods and results

Streptozotocin injection was used to induce diabetes. Diabetic rats received nobiletin orally at 10 or 25 mg/kg daily for 30 days. Diabetic rats displayed significant elevations in glucose, glycosylated hemoglobin (HbA1c), Homeostatic Model of Insulin Resistance (HOMA-IR), and pro-inflammatory cytokines, while the serum levels of insulin, testosterone, luteinizing hormone (LH), and follicle-stimulating hormone (FSH) were significantly reduced. Histological changes to positivity for caspase-3 and decreased androgen receptors (AR) immunoexpression were observed in diabetic rats. Both doses of nobiletin improved hyperglycemia, reduced pro-inflammatory cytokines, and augmented insulin, testosterone, LH, and FSH levels. LH and FSH receptors and cytochrome P450 17 α-hydroxylase (CYP17A1) were markedly downregulated in terms of both gene and protein expression in testicular tissues of the diabetic group, effects that were markedly ameliorated with both doses of nobiletin. In addition, both doses significantly reduced lipid peroxidation and caspase-3 immunoexpression and improved the activity of the antioxidant enzymes and AR in testicular tissues of the diabetic group.

Conclusion

Both nobiletin doses showed protective effects against diabetes-induced testicular injury by reducing oxidative stress, hyperglycemia, inflammation, and caspase-3 and upregulating the hypophysis–gonadal axis and AR. The high dose of nobiletin was more effective than the lower one.

     

Nigella sativa thymoquinone-rich fraction greatly improves plasma antioxidant capacity and expression of antioxidant genes in hypercholesterolemic rats

The antioxidant activities of the thymoquinone-rich fraction (TQRF) extracted from Nigella sativa and its bioactive compound, thymoquinone (TQ), in rats with induced hypercholesterolemia were investigated. Rats were fed a semipurified diet supplemented with 1% (w/w) cholesterol and were treated with TQRF and TQ at dosages ranging from 0.5 to 1.5 g/kg and 20 to 100 mg/kg body wt, respectively, for 8 weeks. The hydroxyl radical (OH^·)-scavenging activity of plasma samples collected from experimental rats was measured by electron spin resonance. The GenomeLab Genetic Analysis System was used to study the molecular mechanism that mediates the antioxidative properties of TQRF and TQ. Plasma total cholesterol and low-density-lipoprotein cholesterol levels were significantly decreased in the TQRF- and TQ-treated rats compared to untreated rats. Feeding rats a 1% cholesterol diet for 8 weeks resulted in a significant decrease in plasma antioxidant capacity, as measured by the capacity to scavenge hydroxyl radicals. However, rats treated with TQRF and TQ at various doses showed significant inhibitory activity toward the formation of OH^· compared to untreated rats. Upon examination of liver RNA expression levels, treatment with TQRF and TQ caused the up-regulation of the superoxide dismutase 1 (SOD1), catalase, and glutathione peroxidase 2 (GPX) genes compared to untreated rats (P < 0.05). In support of this, liver antioxidant enzyme levels, including SOD1 and GPX, were also apparently increased in the TQRF- and TQ-treated rats compared to untreated rats (P < 0.05). In conclusion, TQRF and TQ effectively improved the plasma and liver antioxidant capacity and enhanced the expression of liver antioxidant genes of hypercholesterolemic rats.     

Detection and validation of single feature polymorphisms using RNA expression data from a rice genome array

Background  

A large number of genetic variations have been identified in rice. Such variations must in many cases control phenotypic differences in abiotic stress tolerance and other traits. A single feature polymorphism (SFP) is an oligonucleotide array-based polymorphism which can be used for identification of SNPs or insertion/deletions (INDELs) for high throughput genotyping and high density mapping. Here we applied SFP markers to a lingering question about the source of salt tolerance in a particular rice recombinant inbred line (RIL) derived from a salt tolerant and salt sensitive parent.     

Occurrence of two closely related ricefishes, Javanese medaka (Oryzias javanicus) and Indian medaka (O. dancena) at sites with different salinity in Peninsular Malaysia

Among ricefishes of the genus Oryzias, the Javanese medaka (O. javanicus) and the Indian medaka (O. dancena) are highly adaptable to seawater. Although wide distribution of the two species in the brackish waters of South and South-East Asia has been reported, their habitat preference remains unknown. We surveyed 12 sites in five estuarial areas of the west coast of Peninsular Malaysia from Kuala Gula to Tanjung Piai. Both species were found in all five areas, suggesting their distribution throughout the west coast of Peninsular Malaysia. This is the southernmost-recorded appearance of O. dancena, to the best of our knowledge. However, the habitats of the two species were essentially separated: of the 12 surveyed sites, the species were found in co-existence at only two sites, and one or the other species was found alone at the remaining 10 sites. We compared temperature, salinity, pH, and dissolved oxygen (DO) at the sampling sites and found that the habitat of O. javanicus is with higher salinity and DO. The salinity and DO at the sites of co-existence are near the lowest values found at the O. javanicus-only sites, and the highest values at the O. dancena-only sites. These results suggest that O. javanicus and O. dancena habitats are essentially separated; the former prefers hyperosmotic conditions while the latter prefers hypoosmotic conditions, and the latter may be more tolerant of hypoxia. The two sites of co-existence are points of contact between the species’ separate distribution areas.     

An electrochemical impedance spectroscopy (EIS) assay measuring the calcification inhibition capacity in biological fluids

Pathological calcification of the cardiovascular system is one of the major causes of high mortality and morbidity in dialysis patients. The inhibition of ectopic calcification relies (I) on the formation of calciprotein particles (CPPs), nanospherical complexes of calcium phosphate mineral, fetuin-A and other acidic serum proteins, and (II) on the stabilization of calcium phosphate prenucleation clusters by fetuin-A monomers. In supersaturated serum, mineral ion aggregation leads to a change in the electrical impedance. In this work, we present a method based on electrochemical impedance spectroscopy (EIS) to establish an impedance trace of mineral ion clustering in vitro. In the presence of 20 μM of serum protein fetuin-A, a prototypic calcification inhibitor, we measured a change in impedance (Δ(R)) of 195.52 ± 27.78%Ω compared to 430.41 ± 11.36%Ω in inhibitor-free samples. We also identified a CPP-formation dependency on the actual content of ions and protein in the samples under investigation. Two-step ripening of CPP was also observed. The presented method may form the basis of a simple label-free bedside or online test to be used in routine clinical practice for estimating the calcification risk in serum.     

Spectroscopic studies of 9-hydroxyellipticine binding to DNA

The binding of 9-hydroxyellipticine to calf thymus DNA, poly[d(A-T)]₂, and poly-[d(G-C)]₂ has been studied in detail by means of CD, linear dichroism, resonance light scattering, and molecular dynamics. The transition moment polarizations of 9-hydroxyelliptiycine were determined in polyvinyl alcohol stretched film. Spectroscopic solution studies of the DNA/drug complex are combined with theoretical CD calculations using the final 50 ps of a series of molecular dynamics simulations as input. The spectroscopic data shows 9-hydroxyellipticine to adopt two main binding modes, one intercalative and the other a stacked binding mode involving the formation of drug oligomers in the DNA major groove. Analysis of the intercalated binding mode in poly[d(A-T)]₂ suggests the 9-hydroxyellipticine hydroxyl group lies in the minor groove and hydrogen bonds to water with the pyridine ring protruding into the major groove. The stacked binding mode was examined using resonance light scattering and it was concluded that the drug was forming small oligomer stacks rather than extended aggregates. Reduced linear dichroism measurements suggested a binding geometry that precluded a minor groove binding mode where the plane of the drug makes a 45° angle with the plane of the bases. Thus it was concluded that the drug stacks in the major groove. No obvious differences in the mode of binding of 9-hydroxyellipticine were observed between different DNA sequences; however, the stacked binding mode appeared to be more favorable for calf thymus DNA and poly[d(G-C)]₂ than for poly[d(A-T)]₂, an observation that could be explained by the slightly greater steric hindrance of the poly[d(A-T)]₂ major groove. A strong concentration dependence was observed for the two binding modes where intercalation is favored at very low drug load, with stacking interactions becoming more prominent as the drug concentration is increased. Even at DNA : drug mixing ratios of 70:1 the stacked binding mode was still important for GC-rich DNAs. © 1998 John Wiley & Sons, Inc. Biopoly 46: 127–143, 1998     

Modulatory Effect of Lycopene on Deltamethrin-Induced Testicular Injury in Rats

Aim of the current study was to investigate the ability of deltamethrin to induce testicular injury in rats and its possible attenuation with lycopene. Rats were divided into three groups: Group I (DEL) received deltamethrin, 5 mg/kg b.w./day orally, in corn oil. Group II (DEL + Lyc) received oral dose of lycopene (4 mg/kg b.w./day) in corn oil concurrently with deltamethrin following the same regimen as in group I. Group III (Control) received appropriate volume of corn oil. After 4 weeks, deltamethrin-treated rats showed decreased body weight, serum testosterone, luteinizing hormone and follicle-stimulating hormone levels. Testicular total oxidant capacity (TOC), nitrite/nitrate (NOx), poly (ADP-ribose) polymerase (PARP), and DNA damage were significantly increased. RT-PCR demonstrated significant up-regulation in testicular mRNA for glutathione-S-transferase and heat-shock protein-70 (HSP-70), whereas steroidogenic acute regulatory (StAR) protein was down-regulated after deltamethrin exposure. Lycopene was able to restore body weight, serum testosterone, StAR mRNA, TOC, NOx levels, and PARP activity with significant decrease in HSP-70 mRNA, and DNA damage. In conclusion, lycopene was able to counteract the deleterious effect of deltamethrin.