Molecular Modeling of Disease Causing Mutations in Domain C1 of cMyBP-C

Cardiac myosin binding protein-C (cMyBP-C) is a multi-domain (C0–C10) protein that regulates heart muscle contraction through interaction with myosin, actin and other sarcomeric proteins. Several mutations of this protein cause familial hypertrophic cardiomyopathy (HCM). Domain C1 of cMyBP-C plays a central role in protein interactions with actin and myosin. Here, we studied structure-function relationship of three disease causing mutations, Arg177His, Ala216Thr and Glu258Lys of the domain C1 using computational biology techniques with its available X-ray crystal structure. The results suggest that each mutation could affect structural properties of the domain C1, and hence it’s structural integrity through modifying intra-molecular arrangements in a distinct mode. The mutations also change surface charge distributions, which could impact the binding of C1 with other sarcomeric proteins thereby affecting contractile function. These structural consequences of the C1 mutants could be valuable to understand the molecular mechanisms for the disease.     

Hypothalamic S-Nitrosylation Contributes to the Counter-Regulatory Response Impairment following Recurrent Hypoglycemia

Aims

Hypoglycemia is a severe side effect of intensive insulin therapy. Recurrent hypoglycemia (RH) impairs the counter-regulatory response (CRR) which restores euglycemia. During hypoglycemia, ventromedial hypothalamus (VMH) production of nitric oxide (NO) and activation of its receptor soluble guanylyl cyclase (sGC) are critical for the CRR. Hypoglycemia also increases brain reactive oxygen species (ROS) production. NO production in the presence of ROS causes protein S-nitrosylation. S-nitrosylation of sGC impairs its function and induces desensitization to NO. We hypothesized that during hypoglycemia, the interaction between NO and ROS increases VMH sGC S-nitrosylation levels and impairs the CRR to subsequent episodes of hypoglycemia. VMH ROS production and S-nitrosylation were quantified following three consecutive daily episodes of insulin-hypoglycemia (RH model). The CRR was evaluated in rats in response to acute insulin-induced hypoglycemia or via hypoglycemic-hyperinsulinemic clamps. Pretreatment with the anti-oxidant N-acetyl-cysteine (NAC) was used to prevent increased VMH S-nitrosylation.

Results

Acute insulin-hypoglycemia increased VMH ROS levels by 49±6.3%. RH increased VMH sGC S-nitrosylation. Increasing VMH S-nitrosylation with intracerebroventricular injection of the nitrosylating agent S-nitroso-L-cysteine (CSNO) was associated with decreased glucagon secretion during hypoglycemic clamp. Finally, in RH rats pre-treated with NAC (0.5% in drinking water for 9 days) hypoglycemia-induced VMH ROS production was prevented and glucagon and epinephrine production was not blunted in response to subsequent insulin-hypoglycemia.

Conclusion

These data suggest that NAC may be clinically useful in preventing impaired CRR in patients undergoing intensive-insulin therapy.     

Circulating and tissue microRNAs as a potential diagnostic biomarker in patients with thrombotic events

Venous and arterial thrombosis are conditions that have a considerable burden if left untreated. The hypoxia-induced by the occluded vessel can disrupt the circulation of any organ, the cornerstone of treating thrombosis is rapid diagnosis and appropriate treatment. Diagnosis of thrombosis may be made by using laboratory tests or imaging techniques in individuals who have clinical manifestations of a thrombotic event. The use of serum micro ribonucleic acids (RNAs) has recently been applied to the diagnosis of thrombosis. These small RNA molecules are emerging as new diagnostic markers but have had very limited applications in vascular disease. Most of the articles provided various microRNAs with different levels of accuracy. However, there remains a lack of an appropriate panel of the most specific microRNA in the literature. The purpose of the present review was to summarize the existing data on the use of microRNAs as a diagnostic biomarker for venous thrombosis.     

The survivin molecule as a double-edged sword in cellular physiologic and pathologic conditions and its role as a potential biomarker and therapeutic target in cancer

Survivin is a member of the family of apoptosis inhibitory proteins with increased expression level in most cancerous tissues. Evidence shows that survivin plays regulatory roles in proliferation or survival of normal adult cells, principally vascular endothelial cells, T lymphocytes, primitive hematopoietic cells, and polymorphonuclear neutrophils. Survivin antiapoptotic role is, directly and indirectly, related to caspase proteins and shows its role in cell division through the chromosomal passenger complex. Survivin contains many genetic polymorphisms that the role of some variations has been proven in several cancers. The −31G/C polymorphism is one of the most important survivin mutations which is located in the promoter region on a CDE/CHR motif. This polymorphism can upregulate the survivin messenger RNA. In addition, its allele C can increase the risk of cancers in 1.27-fold than allele G. Considering the fundamental role of survivin in different cancers, this protein could be considered as a new therapeutic target in cancer treatment. For this purpose, various strategies have been designed including the prevention of survivin expression through inhibition of mRNA translation using antagonistic molecules, inhibition of survivin gene function through small inhibitory molecules, gene therapy, and immunotherapy. In this study, we describe the structure, played roles in physiological and pathological states and genetic polymorphisms of survivin. Finally, the role of survivin as a potential target in cancer therapy given challenges ahead has been discussed.     

The effect of different salts (heavy metals) on the mycelium growth of Nematophagus fungi

Environmental pollution in addition to direct damage on plant growth, with the destruction of biological control agents, causes indirect damage to plants. The aim of this research was to study the effects of different concentrations (0, 500, 1000, 1500 and 2000 ppm) of heavy metals including Ag, Co, Cu, Fe, Hg, Mn, Pb and Zn on the mycelial growth and to assess the fungicidal or fungistatic effects of these salts on five Nematophagus fungi including Trichoderma harzianum (T8), Trichoderma virens (T21), Trichoderma hamatum (T9), Pochonia chlamydosporia var. chlamydosporia and Arthrobotrys oligospora. The results show that Ag, Co, Cu, Fe and Hg could stop the mycelium growth of all fungi, but Mn, Pb and Zn cannot inhibit the growth of these fungi completely. Among the first group, Hg and Cu stopped the growth of fungi even in 500 ppm. Among these metals that inhibit the growth of fungi, Cu has fungistatic effect and others have fungicide effect. The experiment was conducted in vitro condition, using potato dextrose agar (PDA) under complete randomised design with four replications. The data of mycelium growth were recorded at seven days after inoculation at 25 ± 2°C.     

Direct effect of biocontrol agents on wilt and root-rot diseases of sesame

In Egypt, sesame cultivation is subject to attack by wilt and root-rot diseases caused by Fusarium oxysporum f.sp. sesami (Zap) Cast. and Macrophomina phaseolina (Maubl) Ashby causing losses in quality and quantity of sesame seed yield. Bacillus subtilis and Trichoderma viride isolates which were isolated from sesame rhizosphere were the most effective to antagonise fungal pathogens, causing high reduction of hyphal fungal growth. Trichoderma viride was found to be mycoparasitic on Fusarium oxysporum f.sp. sesami and M. phaseolina causing morphological atternation of fungal cells and sclerotial formation. In general, Bacillus subtilis, T. viride, avirulent Fusarium oxysporum isolate and Glomus spp. (Amycorrhizae) significantly reduced wilt and root-rot incidence of sesame plants at artificially infested potted soil by each one or two pathogens. Data obtained indicate that Glomus spp significantly reduced wilt and disease severity development on sesame plants followed by T. viride. Meanwhile, avirulent Fusarium oxysporum isolate followed by Glomus spp. were effective against root-rot disease incidence caused by M. phaseolina. Glomus spp. followed by B. subtilis significantly reduced wilt and root-rot disease of sesame plants. All biotic agents significantly reduced F. oxysporum f.sp. sesami and M. phaseolina counts in sesame rhizosphere at the lowest level. Glomus spp. and the avirulent isolate of F. oxysporum eliminated M. phaseolina in sesame rhizosphere. Meanwhile T. viride was the best agent at reducing F. oxysporum at a lower level than other treatments. Application of VA mycorrhizae (Glomus spp.) in fields naturally infested by pathogens significantly reduced wilt and root-rot incidence and it significantly colonised sesame root systems and rhizospheres compared to untreated sesame transplantings.     

Detection of begomovirus associated with okra leaf curl disease

Leaf curl and yellow vein mosaic viral disease is the major constraint on okra (Abelmoschus esculentus L.) production in India. Amplified fragment sequence of DNA-β showed highest similarity of 91.7% with Bhendi yellow vein mosaic virus-Tamil Nadu (AJ308425, NC_003405) and lowest similarity of 48.5% with OKLCV (NC_004093), whereas coat protein specific amplified sequence showed highest homology with isolate of Madurai, Haryana, Ludhiana and lowest homology of 92% with Mesta yellow vein mosaic Bahraich virus (MYVMBV) (EU360303). The results obtained in the present study confirm that both the viral diseases of okra reported in southern India are caused by a begomovirus associated with DNA-β in which the plants show leaf curl symptoms and never develops yellow vein mosaic and those plants which show yellow vein mosaic, never develops leaf curl symptoms even in the same rows and field. The okra leaf curl is an emerging virus disease in India.     

PTPN22 gene polymorphism in Egyptian alopecia areata patients and its impact on response to diphencyprone immunotherapy

PTPN22 1858C>T gene polymorphism has been associated with several autoimmune disorders including alopecia areata. The aim of the current study was to investigate the effect of the inherited genetic polymorphism 1858C>T of PTPN22 gene on the predisposition to severe forms of alopecia areata and its effect on the response to DPC treatment. To achieve our aim, PTPN22 1858C>T genotyping was performed by PCR-based restricted fragment length polymorphism (PCR-RFLP) analysis. The study included 103 Egyptian patients with extensive alopecia areata treated by DPC. Hundred healthy age and sex matched blood donors were included in the current study as a control group. Results of genotyping showed that PTPN22 CT and TT mutant genotypes were significantly higher in AA patients compared to controls and conferred increase risk of AA (OR = 2.601, 95% CI = 1.081–6.255). Statistical comparison between AA patients with wild and mutant genotypes revealed that the duration of the illness was significantly longer in those harboring the mutant genotypes. Moreover, the association of other autoimmune diseases as atopy and diabetes mellitus was higher in patients with mutant genotypes. Furthermore, PTPN22 1858C>T genetic polymorphism did not affect the patients' response to DPC immunotherapy.     

Analysis of CLDN14 gene in deaf Moroccan patients with non-syndromic hearing loss

Mutations in the CLDN14 gene, encoding the tight junction claudin 14 protein has been reported to date in an autosomal recessive form of isolated hearing loss DFNB29. In order to identify the contribution of CLDN14 to inherited deafness in Moroccan population, we performed a genetic analysis of this gene in 80 Moroccan familial cases. Our results show the presence of 7 mutations: 6 being conservative and one leading to a missense mutation (C11T) which was found at heterozygous and homozygous states, with a general frequency of 6.87%. The pathogenicity of the resulting T4M substitution is under discussion.