In silico elucidation of potential drug targets against oxygenase domain of Human eNOS Dysfunction.

Nitric Oxide (NO) signaling pathway plays a vital role in various physiological and pathophysiological processes including vasodilation, neurogenesis, inflammation, translation and protein regulation. NO signaling pathway is associated with various diseases such as cardiovascular diseases, vision impairment, hypertension and Alzheimer's disease. Human Endothelial Nitric Oxide Synthase (eNOS) bound with calcium regulatory protein (calmodulin (CaM)) to produce NO which initiates cGMP pathway. The current study employs to screen the novel compounds against human eNOS independent of calcium regulatory protein (CaM). The current effort emphasized that the deficiency of CaM leads to dysfunction of cGMP signaling pathway. In this work, a hybrid approach of high-throughput virtual screening and comparative molecular docking studies followed by molecular dynamic simulation analyses were applied. The screening of top ranked two novel compounds against eNOS were reported that showed effective binding affinity, retrieved through the DrugBank and ZINC database libraries. Comparative molecular docking analyses revealed that Val-104, Phe-105, Gln-247, Arg-250, Ala-266, Trp-330, Tyr-331, Pro-334, Ala-335, Val-336, Tyr-357, Met-358, Thr-360, Glu-361, Ile-362, Arg-365, Asn-366, Asp-369, Arg-372, Trp-447 and Tyr-475 are potent residues for interactional studies. High-throughput virtual screening approach coupled with molecular dynamic simulation and drug likeness rules depicted that ZINC59677432 and DB00456 are potent compounds to target eNOS. In conclusion, the proposed compounds are potent against eNOS based on extensive in silico analyses. Overall, the findings of this study may be helpful to design therapeutic targets against eNOS.

Domain analyses of Usher syndrome causing Clarin-1 and GPR98 protein models.

Usher syndrome is an autosomal recessive disorder that causes hearing loss, Retinitis Pigmentosa (RP) and vestibular dysfunction. It is clinically and genetically heterogeneous disorder which is clinically divided into three types i.e. type I, type II and type III. To date, there are about twelve loci and ten identified genes which are associated with Usher syndrome. A mutation in any of these genes e.g. CDH23, CLRN1, GPR98, MYO7A, PCDH15, USH1C, USH1G, USH2A and DFNB31 can result in Usher syndrome or non-syndromic deafness. These genes provide instructions for making proteins that play important roles in normal hearing, balance and vision. Studies have shown that protein structures of only seven genes have been determined experimentally and there are still three genes whose structures are unavailable. These genes are Clarin-1, GPR98 and Usherin. In the absence of an experimentally determined structure, homology modeling and threading often provide a useful 3D model of a protein. Therefore in the current study Clarin-1 and GPR98 proteins have been analyzed for signal peptide, domains and motifs. Clarin-1 protein was found to be without any signal peptide and consists of prokar lipoprotein domain. Clarin-1 is classified within claudin 2 super family and consists of twelve motifs. Whereas, GPR98 has a 29 amino acids long signal peptide and classified within GPCR family 2 having Concanavalin A-like lectin/glucanase superfamily. It was found to be consists of GPS and G protein receptor F2 domains and twenty nine motifs. Their 3D structures have been predicted using I-TASSER server. The model of Clarin-1 showed only α-helix but no beta sheets while model of GPR98 showed both α-helix and ß sheets. The predicted structures were then evaluated and validated by MolProbity and Ramachandran plot. The evaluation of the predicted structures showed 78.9% residues of Clarin-1 and 78.9% residues of GPR98 within favored regions. The findings of present study has resulted in the three dimensional structure prediction and conserved domain analysis which will be quite beneficial in better understanding of molecular components, protein-protein interaction, clinical heterogeneity and pathophysiology of Usher syndrome.

Blunt thoracic trauma--an analysis of 264 patients in Rawalpindi, Pakistan.

OBJECTIVE: To analyse the outcome in terms of morbidity and mortality in blunt thoracic trauma patients in tertiary care hospitals, Rawalpindi. METHODS: The prospective study was conducted from March 2008 to February 2012 in surgical wards of public and private sector hospitals in Rawalpindi. A total of 221 patients were included from the Combined Military Hospital during 2008-10, and 43 patients from the Heart's International during 2011-12. The patients reported to emergency department within 48 hours of trauma. All patients were subjected to detailed history and respiratory system examination to ascertain fracture of ribs, flail segment and haemopneumothorax. The diagnosis of chest wall injuries, parenchymal pulmonary injuries and pleural involvement were made on the basis of chest radiographs and computed tomography scan of the chest. The lung contusion was assessed by the number of lobes involved. SPSS 19 was used for statistical analysis. RESULTS: Of the 264 patients in the study, 211 (80%) were males and 54 (20%) females. The overall mean age was 44.8 +/- 17.1 years. Over all morbidity was 222 (84.2%); morbidity (minor) was 128 (48.5%), and morbidity (major) was 94 (35.7%). Mortality was 26 (9.8%) and 16 (6%) cases had normal outcome. CONCLUSION: Early identification and aggressive management of blunt thoracic trauma is essential to reducing significant rates of morbidity and mortality.

Comparison of outcome between low and high thoracic trauma severity score in blunt trauma chest patients.

BACKGROUND: Blunt chest trauma is second leading cause of death among trauma patients. Early identification and aggressive management of blunt thoracic trauma is essential to reduce the significant rates of morbidity and mortality. Thoracic trauma severity score (TTS) is a better predictor of chest trauma related complications. The objective of the study was to compare outcomes between low-and high thoracic trauma severity score in blunt trauma chest patients. METHODS: A cross sectional descriptive study was carried out in public and private sector hospitals of Rawalpindi, Pakistan from 2008 to 2012 and 264 patients with blunt trauma chest who reported to emergency department of the hospitals, within 48 hrs of trauma were recruited. All patients were subjected to detailed history and respiratory system examination to ascertain fracture ribs, flail segment and hemopneumothorax. Written and informed consent was taken from each patient. Permission was taken from ethical committee of the hospital. RESULTS: The patients with blunt chest trauma had an array of associated injuries; however there were 70.8% of patients in low TTS group and 29.2% in high TTS group. Outcome was assessed as post trauma course of the patient. Outcome in low and high TTS group was compared using Chi square test which shows a significant relationship (p=0.000) between outcome and TTS, i.e., outcome worsened with increase in TTS. CONCLUSION: It is concluded that there is a significant relationship between outcome and thoracic trauma severity. Outcome of the patient worsened with increase in thoracic trauma severity score.

Screening and design of anti-diabetic compounds sourced from the leaves of neem (Azadirachta indica).

Diabetes Mellitus is affecting people of all age groups worldwide. Many synthetic medicines available for type 2 diabetes mellitus in the market. However, there is a strong requirement for the development of better anti-diabetes compounds sourced especially from natural sources like medicinal plants. The extracts from the leaves of neem (Azadirachta indica) is traditionally known to have anti-diabetes properties. Therefore, there is an increased interest to identify potential compounds identified from neem leaf extracts showing predicted binding property with the known diabetes mellitus type 2 protein enzyme target phosphoenol-pyruvate carboxykinase(PEPCK). The structure data for compounds found in the leaf extract of neem was screened against PEPCK using molecular docking simulation and screening techniques. Results show that the compound 3-Deacetyl-3-cinnamoyl-azadirachtin possesses best binding properties with PEPCK. This observation finds application for further consideration in in vitro and in vivo validation.

An unusual case of Youssef's syndrome (vesicouterine fistula) and its relationship with placenta percreta.

A pregnant lady, on antenatal ultrasound examination, was found to have placenta percreta involving the bladder. After cesarean section, she developed the typical triad of Youssef's syndrome i.e. cyclic hematuria and amenorrhea without vaginal leakage of urine. Cystoscopy showed a vesicouterine fistula which was treated through the transabdominal approach.