Volume 5, Issue 3

 Completely sequenced organisms have some uncharacterized proteins that are gene-encoded products. These proteins can be predicted through in-silico approaches and their biological activities are not proved by experimental evidence and known as hypothetical proteins (HPs). These proteins are important due to their excessive involvement in different cellular and signaling pathways. Structural and functional characterization of HPs reveal crucial roles in microorganisms, especially in pathogens related to human diseases. Here, we discussed all possibilities of in-silico analysis tools and other recently reported methods for hypothetical protein characterization and biomedical applications, including drug and vaccine development. Different methodologies, including meta-proteomics have been used to study protein expression by identification of HPs and comparative genomics have also come under observation due to the emergence of evolutionary study among different organisms. Structural characterization of proteins acts as a base for their functional prediction, novel drug target identification for disease treatment, vaccine production and sero-diagnosis. HPs have played major roles in different vital phenomenon for life including host adaptation, wound healing and chemotaxis. In the current era of drug and antibiotic resistance, HPs can be novel targets to treat related diseases. Identification and characterization of most HPs are under observation and will be the most promising genomic and bioinformatics techniques in structure-based drug designing and vaccine production in future. 

 Background: The spread of Zika virus is of great concern as it is recently becoming the third global infectious disease outburst after H1N1 flu and the Ebola virus infections. Centre for Disease Control (CDC) categorized Pakistan, India, and Bangladesh as countries vulnerable to Zika Risk. Realizing health implications of this emerging epidemic, it is a dire need to build an all-inclusive view of the status of research on Zika virus disease, and a lucid picture of the research output and scientific collaborations in the field. Methods: All the articles published globally on Zika virus during 2008-2017 and documented in Web of Science were analyzed using Microsoft Excel and Word Cloud tool. The data were extracted from all databases of the Web of Science, obtaining a total of 3384 articles for analysis. Results: 3384 records on Zika virus research were indexed in the Web of Science database during 2008-2017. The retrieved data indicate that over the past ten years, not much research has been done on this virus and the focus shifted to research on Zika in the last three years only and the number of researches increased from just 38 in 2015 to 1962 in 2017. Pakistan has a low share in global publications on Zika with a total number of 24 publications. “Honein Margaret” is considered the most active researcher in the field, by contributing to 80 articles. Most of the published research on Zika virus is from US (47.07%). Conclusion: When compared with other countries, the contribution of Pakistan is negligible with a global share of 0.71% on Zika virus. Serious focus on research is needed in this field realizing the severe medical, ethical, and economic implications of this emerging epidemic in Pakistan. 

 Background: Lead is most commonly released environmental contaminant making its way to air, soils and water. It causes hormonal imbalance and over production of reactive oxygen in plants when absorbed through leaves and roots. It contaminates the ground water depending on the type of soils and characteristics of lead. Plants ability to tolerate lead is linked with cell wall potential, activation of antioxidants defense mechanism and synthesis of osmolytes. Methods: The study was designed to evaluate the effects of Pb(NO₃)₂ induced stress on biological activity and bioactive compounds in maize. The plants were subjected under two different lead concentrations (T1- 0.35mg/ml and T2- 0.45mg/ml). Phytochemical screening revealed the presence of alkaloids, coumarins, saponins, tannins and terpenoids in maize. Total Phenolic Content (TPC) was increased (T1- 45%, T2- 58.42%) under lead stress when compared with control (36.29%). The cytotoxicity was checked using hemolytic activity against human red blood cells. Results: The scavenging rate was highest (T1- 33.5%, T2- 52%) when compared with control (18.6%). Zone of inhibition of Aspergillus niger was highest amongst other fungal strains. The HPLC results showed that maize has some phyto-ingredients which may be accountable for cell reinforcement and anti-microbial activity. The extracts were further analyzed for the biochemical profile like superoxide dismutase, peroxidase, catalase, amylase and protease. Escherichia colishowed maximum activity with control (25±3.46mm) and maximum under stress (T1- 17±1.633 mm, T2- 20±4.08 mm). Conclusion: Lead stress altered all the activities when compared to control plants. In conclusion, Maize can be used as a potential indicator for lead and other compounds to play a vital role in phytoremediation. The results would further lead to find the new compounds and plant mechanism to cope with stress. 

 Background: Tramadol-diclofenac (TD-DF) could be used in chronic pain management. Concurrent use may present renal complications due to their individual nephrotoxic profile. The present study assessed the kidney function and histology of tramadol-diclofenac treated albino rats. Methods: Forty two adult albino rats divided into seven groups A-G were used for this study. Rats were orally administered with TD (12 mg/kg/day), DF (6 mg/kg/day), and TD-DF for 14 days including two recovery groups. Rats were weighed and sacrificed at the termination of drug treatment. Serum was extracted from blood and evaluated for creatinine (Cr), urea (U), uric acid (UA), total protein (TP), albumin (Ab) and serum electrolytes (K⁺, Na⁺, Cl^-, and HCO3^-). Kidneys were excised weighed and evaluated for alanine aminotransferase (ALT), alkaline phosphatase (ALP), aspartate aminotransferase (AST), superoxide dismutase (SOD), catalase (CAT) glutathione (GSH), glutathione peroxidase (GPX), malondialdehyde (MDA) levels and histological damage. Results: The body weight, absolute and relative kidney weights and serum electrolytes were not significantly (p> 0.05) altered in the TD-DF treated rats in comparison to control. However, the levels of Cr, U, UA, AST, ALT, ALP and MDA were significantly (p<0.05) increased whereas Ab, TP, SOD, GSH, GPX and CAT were significantly (p<0.05) decreased in the TD-DF treated rats in comparison to treatments with individual doses of TD and DF. Varying degrees of histological damage were observed in the kidneys of TD-DF treated rats. However, nephrotoxic effects due to treatment with TD-DF were reversed in the recovery groups. Conclusion: The use of tramadol-diclofenac could be associated with reversible nephrotoxicity; therefore renal function assessment is advised before tramadol-diclofenac use. 

 Background: Edible mushrooms are considered as the significant source of minerals and vitamins.Lactarius deliciosus and Pleurotus ostreatus are the two known edible mushroom species because of their distinctive taste as well as nutritional properties. Methods: L. deliciosus and P. ostreatus were collected from the native flora of Amanos Mountains in Turkey. The methanol (MeOH) extract of the L. deliciosus and P. ostreatus was obtained to assess antioxidant potential, Total phenolic, flavonoid, β-carotene and lycopene content. Results: Total phenolic (mg/kg), flavonoid (mg/kg) and β-carotene (mg/100 ml) contents of the MeOH extracts of L. deliciosus and P. ostreatus were 34.55, 6.03, 1.15 and 15.66, 0.16, 0.02, respectively. Lycopene was only detected in L. deliciosus with a content of 0.25 (mg/100 ml). At 5 mg/ml, DPPH (%), RP (Abs.), H₂O₂ (%), NO (%) and FRAP (mmol Fe²⁺/L) activities of L. deliciosusand P. ostreatus were 45.33, 0,790, 88.30, 55.51, 0.57 and 17.42, 0,530, 73.77, 33.64, 0.28, respectively. Conclusion: Pearson correlation among the antioxidant results was found to be well correlated (r ≥0.90).  Antioxidant results were highly correlated (r ≥0.93) with total phenolic compounds and lycopene.  As of date, there has no previous work not only the NO but also H₂O₂ radical scavenging capabilities of L. deliciosus and P. ostreatus. 

 Background: Fluorescent marker genes have modernized many areas of molecular biology, specifically in plant biotechnology and genetic engineering studies. The use of fluorescent proteins permits the scientists to purify the desired clones visually in transformation work. Green fluorescent protein (gfp) derived from Aequorea victoria has been the most common and favorite fluorescent marker which is being widely used as a visual selection marker gene. It can be easily visualized under UV light without the involvement of any substrate and is non-destructive as well. Method: A species-specific chloroplast transformation vector was constructed with gfp as a fluorescent marker gene. The recombinant vector was biolistically integrated in tobacco plastome and transgenic cells were initially screened on spectinomycin containing regeneration medium. Results: The successful plastome integration was verified by using cellular DNA from drug resistant clones in PCR and southern blotting. The expression of gfp in transplastomic clones was microscopically investigated using simple florescent as well as confocal laser scanning microscopes. Conclusion: Regeneration of transgenic plants was significantly helped by visual identification of fluorescent at different stages of development, also enabling to identify the homozygous and heterozygous tissues. No toxic effect of the gfp was observed and lack of toxicity as maintained by normal phenotypic performance of plants. 

 Background: Anesthetics are responsible for imparting many effects on the central nervous system. We hypothesized that short duration anesthetics may have varied effects at different time of the day. Propofol (short duration anaesthesia) has showed circadian variation in loss of righting reflex during 24 hours of the day. Methods: Characterization of the effects of propofol anaesthesia on ERK1/2 in hippocampus at two different times of the day was performed. Male rats received either an intra-peritoneal injection of 120 mg/kg of propofol to set short-duration anaesthesia state (20-30 minutes) or the equivalent amount of the lipidic solvent as controls. For both groups of animals, anaesthesia or control, the injections were performed either at ZT0 or at ZT12. One hour following the injection, the animals were euthanized; the brains were removed and immediately frozen. The amount of ERK1/2 was assessed by using immunohistochemistry on brain sections. Results: The amount of ERK1/2 density was significantly decreased (P<0.05) in CA1, CA2 and CA3 areas of the hippocampus only when anaesthesia was performed at ZT12. Conclusion: Our current results evidenced that the impact of propofol anaesthesia on hippocampus vary depending on the zeitgeber time. 

 Background: Molecular characterization of MAOA gene was performed to investigate aggressive behaviour within wild (lion, leopard, and wolf) as opposed to docile animal (sheep, goat) species living in different habitats, by undertaking sequence diversity analysis of this gene. Methods: The MAOA gene was partially amplified by PCR for wild and docile animal species. Amplified DNA was sequenced and then analyzed using BioEdit and Sequencher softwares, while multiple sequence alignment and phylogenetics analysis were conducted through MEGA software. Bioinformatics tool like Prosite scan, Motif Scan and Prot Param were used to study properties of mutant proteins of MAOA gene. Results: Different polymorphic sites were observed which included c.956, c.1063 in docile animals and c.2530 in wild animals. Phylogenetic analysis based on this candidate gene endorsed the existing taxonomy of subject animals, while bioinformatics tools explored the altered characteristics of mutant MAOA protein.  Conclusion: The newly found polymorphic loci in wild and docile animals in this study could have a role in behavioral response and acclimatization within their peculiar habitats. This study also highlights the genetic diversity of MAOA gene, which will add knowledge to the existing animal genetic resource of Pakistan.