In Silico identification of novel phytochemicals that target SFRP4: An early biomarker of diabesity.

The simultaneous coexistence of complicated metabolic conditions like obesity and diabetes within an individual is known as diabesity. Obesity is the key factor for many chronic diseases, including insulin resistance and type 2 diabetes (T2D). Metabolic stress due to nutrient overload releases different inflammatory mediators. Secreted frizzled-related protein 4 (SFRP4) is also an inflammatory mediator that impairs insulin secretion. SFRP4 acts as an early biomarker for diabesity expressed with interleukin-1 beta (IL-1ß) in the adipose tissues that hinder the exocytosis of insulin-secreting granules from the pancreatic ß-cells and is a potential target for preserving ß-cell dysfunction and the diabesity treatment. The current study aimed to screen potential bioactive compounds targeting and inhibiting the diabesity-linked SFRP4 protein through an in silico approach. The three-dimensional (3D) structure of human SFRP4 was predicted through comparative modeling techniques and evaluated by various online bioinformatics tools. The molecular docking and MD simulation investigations were carried out against phytochemicals with anti-diabetic and anti-obesity properties to shortlist the best SFRP4 inhibitor. Hesperetin, Curcumin, Isorhamnetin, Embelin, Epicatechin, and Methyl Eugenol interacted strongly with SFRP4 by displaying zero RMSD and binding affinities of -6.5, -6.4, -6.3, -5.3, -6.3 and -5.8 kcal/mol respectively. Additionally, the root mean square fluctuation and root mean square deviation graphs from the MD simulation results demonstrated that hesperetin has good variations throughout the simulation period as compared to others. This dynamic stability and control behavior of hesperetin, when it interacts with SFRP4, shows that it has the potential to modulate the function and activity of the protein. Therefore, hesperetin is identified as an effective and top drug candidate through this analysis for preserving beta-cell function and treating diabesity by targeting SFRP4. The findings of this study could be useful in the design and development of diabesity drugs.

HPLC-DAD analysis of Quercus leucotrichophora extract and appraisal of its antiasthmatic potential via modulation of aquaporins, inflammatory, and oxidative stress biomarkers in Albino mice.

Herbal drugs offer an alternative approach for the treatment of diseases like asthma due to low cost and comparatively less adverse effects in contrast to synthetic drugs. Leaves of Quercus leucotrichophora are traditionally used for the treatment of asthma. The study was aimed to assess the anti-asthmatic activity of Quercus leucotrichophora (QL) methanolic (QLME) and aqueous extracts (QLAE) in ovalbumin-(OVA) induced asthma and chemical characterization of QL extract by High Performance Liquid Chromatography-Diode array detector (HPLC-DAD). Animals were inoculated with OVA (i.p) on day 1 and 14 followed by intranasal challenge on 27th and 29th day. Both extracts of QL at 600, 300 and 150 mg/kg and dexamethasone (2 mg/kg) l were administered consecutively from days 15-26 via oral gavage. The QL extracts notably reduced (p < 0.0001-p < 0.05) total and differential leukocyte counts in blood and BALF and serum IgE levels in contrast to disease control. Both extracts and Dex substantially improved activities of superoxide dismutase, catalase, and GSH, while reduced malondialdehyde level in treated mice. Treatment with extracts and Dex caused significant (p < 0.0001-p < 0.05) downregulation of tumor necrosis factor-α, interleukin-4, - 5, - 13, - 6, - 1ß, and NF-κB whereas, increased expression of Aquaporin (AQP) 1 and AQP5 in contrast to disease control. It was inferenced from findings that both extract of QL exhibited notable antiasthmatic potential might be due to presence of Daidzein-glucuronic acid, 3-Hydroxyphloretin 6'-hexoside, Catechin, Quercetin, and Kaemferol.

Identification of Ascorbic Acid and Gallic Acid as Novel Inhibitors of Secreted Frizzled-Related Protein for the Treatment of Obesity-Induced Type 2 Diabetes.

Type 2 diabetes mellitus (T2D) has been reported as major public health issue rising at an alarming rate worldwide, and obesity is the leading risk factor for the development of T2D. Secreted frizzled-related protein 4 (SFRP4) released with inflammatory mediators from adipose tissues constrains the exocytosis of insulin containing granules from the pancreatic islets that leads towards the development to T2D. The significant overexpression of SFRP4 in diabetic patients and its involvement in islet dysfunction suggest its critical role in the development of diabetes. Thus, this study was designed to explore the potential of ascorbic acid (AA) and gallic acid (GA) against SFRP4 for the treatment of diabetes. Molecular docking approach was used for the prediction of binding interactions of AA and GA at the active pocket of SFRP4. Docking analysis indicated strong binding interactions of AA and GA to the amino acid residues at the active site of SFRP4. A significant reduction in the level of SFRP4 was observed in transfected cells treated with AA and GA. For the evaluation of the cytotoxicity of AA and GA against HepG2 cells, MTT assay was performed. The results of MTT assay demonstrated that AA and GA are non-cytotoxic towards HepG2 cells at concentration of 15 µM. The oral administration of AA and GA to diet-induced obese mice caused significant reduction in body weight, blood glucose level, and SFRP4 expression. The results of this study suggest that AA and GA have potential for the treatment of obesity-induced T2D.

Antioxidant and antibacterial activities of Artemisia absinthium and Citrus paradisi extracts repress viability of aggressive liver cancer cell line.

BACKGROUND: Numerous reports show that herbal medicines can be utilized in the treatment of different liver disorders. In this study, antioxidant, antibacterial, and anticancer activities of individual as well as combined 80% ethanolic extracts of Artemisia absinthium leaves and Citrus paradisi peels were investigated. METHODS AND RESULTS: Values of total phenolic contents (TPC), total flavonoid contents (TFC), DPPH-radical scavenging activity, and ferric reducing antioxidant power (FRAP) were measured to explore the antioxidant capacity. To assess antibacterial activity, four bacterial strains (Escherichia coli, Staphylococcus aureus, Salmonella enterica, and Klebsiella pneumoniae) were used. Anticancer activity was assessed on Huh-7 (liver cancer) and Vero (non-cancerous) cell lines. FRAP activity of combined plants extract was higher as compared to their individual effect; the trend did not hold in the case of DPPH-radical scavenging activity. Antibacterial activity of combined extracts by disk diffusion method was observed only against E.coli. MTT results indicated that both plants had a cytotoxic effect on Huh-7 cell line but did not show any effect on Vero cell line. Our data showed a strong negative correlation between the amount of TPC, TFC, & DPPH radicals-scavenging activity and viability of Huh-7 cell line.However, no effect was shown on the non-cancerous cell line. CONCLUSION: The ethanolic extracts of Artemisia absinthium leaves and Citrus paradisi peels can be used against liver cancer because of their antioxidant, antibacterial, and anticancer activities.

Phylogenetic analyses, protein modeling and active site prediction of two pathogenesis related (PR2 and PR3) genes from bread wheat.

Wheat is a major staple food and has been extensively grown around the globe. Sessile nature of plants has exposed them to a lot of biotic and abiotic stresses including fungal pathogen attack. Puccinia graminis f.sp. tritici causes stem rust in the wheat crop and leads to 70% decrease in its production. Pathogenesis-related (PR) proteins provide plants with defense against different fungal pathogens as these proteins have antifungal activities. This study was designed to screen Pakistani wheat varieties for PR2 and PR3 proteins and their in silico characterization. PR2 and PR3 genes were screened and isolated by PCR amplification from wheat variety Chenab-70 and Frontana, respectively. The nucleotide sequences of PR2 and PR3 genes were deposited in GenBank with accession numbers MT303867 and MZ766118, respectively. Physicochemical properties, secondary and tertiary structure predictions, and molecular docking of protein sequences of PR2 and PR3 were performed using different bioinformatics tools and software. PR2 and PR3 genes were identified to encode ß-1,3-glucanase and chitinase proteins, respectively. Molecular docking of both PR2 and PR3 proteins with beta-glucan and chitin (i.e. their respective ligands) showed crucial amino acid residues involved in molecular interactions. Conclusively, molecular docking analysis of ß-1,3-glucanase and chitinase proteins revealed crucial amino acid residues which are involved in ligand binding and important interactions which might have important role in plant defense against fungal pathogens. Moreover, the active residues in the active sties of these proteins can be identified through mutational studies and resulting information might help understanding how these proteins are involved in plant defense mechanisms.

Mangifera indica Extracts as Novel PKM2 Inhibitors for Treatment of Triple Negative Breast Cancer.

Pyruvate kinase (PK), a key enzyme that determines glycolytic activity, has been known to support the metabolic phenotype of tumor cells, and specific pyruvate kinase isoform M2 (PKM2) has been reported to fulfill divergent biosynthetic and energetic requirements of cancerous cells. PKM2 is overexpressed in several cancer types and is an emerging drug target for cancer during recent years. Therefore, this study was carried out to identify PKM2 inhibitors from natural products for cancer treatment. Based on the objectives of this study, firstly, plant extract library was established. In order to purify protein for the establishment of enzymatic assay system, pET-28a-HmPKM2 plasmid was transformed to E. coli BL21 (DE3) cells for protein expression and purification. After the validation of enzymatic assay system, plant extract library was screened for the identification of inhibitors of PKM2 protein. Out of 51 plant extracts screened, four extracts Mangifera indica (leaf, seed, and bark) and Bombex ceiba bark extracts were found to be inhibitors of PKM2. In the current study, M. indica (leaf, seed, and bark) extracts were further evaluated dose dependently against PKM2. These extracts showed different degrees of concentration-dependent inhibition against PKM2 at 90-360 µg/ml concentrations. We have also investigated the anticancer potential of these extracts against MDA-MB231 cells and generated dose-response curves for the evaluation of IC50 values. M. indica (bark and seed) extracts significantly halted the growth of MDA-MB231 cells with IC50 values of 108 µg/ml and 33 µg/ml, respectively. Literature-based phytochemical analysis of M. indica was carried out, and M. indica-derived 94 compounds were docked against three binding sites of PKM2 for the identification of PKM2 inhibitors. The results of in silico based screening have unveiled various PKM2 modulators; however, further studies are recommended to validate their PKM2 inhibitory potential via in vitro biochemical assay. The results of this study provide novel findings for possible mechanism of action of M. indica (bark and seed) extracts against TNBC via PKM2 inhibition suggesting that M. indica might be of therapeutic interest for the treatment of TNBC.

Investigation of Hypoglycemic Peptides Derived from Conserved Regions of adMc1 to Reveal Their Antidiabetic Activities.

Diabetes mellitus is the most common chronic disorder and leading cause of renal, neurological, and gastrointestinal manifestations in developed and developing countries. Despite of many drugs and combinational therapies, the complications of diabetes are still listed due to severe consequences of those drugs. In past few years, plant-derived drugs draw special attention due to their higher efficacy and fewer side-effects. Momordica charantia also known as bitter melon is referred as an antidiabetic and hypoglycemic plant in native populations of Asia and East Africa. In current study, an in silico approach was used to evaluate the interactions and binding patterns of plant-derived peptides devised from a hypoglycemic protein adMc1 of M. charantia as potential inhibitor of DPP-IV, SGLT1, and GLUT2 receptor proteins. The study has described a novel approach to investigate hypoglycemic peptides to cure diabetes. A total of eighty tetra-, penta-, and hexapeptides were devised from conserved regions of adMc1 homologs. The molecular docking approach using MOE software was employed to reveal inhibiting potentials of devised peptides against three selected proteins. Out of 30 shortlisted ligands six peptides (i.e. SMCG, DECC, TTIT, RTTI, ARNL and TVEV) accomplished the criteria of being good drug candidates against selected receptor proteins following the drugability assessment test. The overall results are acceptable on the basis of ADMET profiling for being good drug candidates against selected proteins.

Pathogenic variants of AIPL1, MERTK, GUCY2D, and FOXE3 in Pakistani families with clinically heterogeneous eye diseases.

Significant number out of 2.2 billion vision impairments in the world can be attributed to genetics. The current study is aimed to decipher the genetic basis of Leber congenital Amaurosis (LCA), Anterior Segment dysgenesis (ASD), and Retinitis Pigmentosa (RP), segregating in four large consanguineous Pakistani families. The exome sequencing followed by segregation analysis via Sanger sequencing revealed the LCA phenotypes segregating in families GCUF01 and GCUF04 can be attributed to c.465G>T (p.(Gln155His)) missense and novel c.139_140delinsA p.(Pro47Trhfster38) frameshift variant of AIPL1 and GUCY2D, respectively. The c.1843A>T (p.(Lys615*) truncating allele of MERTK is homozygous in all the affected individuals, presumably suffering with RP, of the GCUF02 family. Meanwhile, co-segregation of the ASD phenotype and the c.289A>G (p.(Ile97Val)) variant of FOXE3 was found in the GCUF06 family. All the identified variants were either absent or present in very low frequencies in the control databases. Our in-silico analyses and 3D molecular modeling support the deleterious impact of these variants on the encoded proteins. Variants identified in MERTK, GUCY2D, and FOXE3 were categorized as "pathogenic" or "likely pathogenic", while the missense variant found in AIPL1 was deemed to have "uncertain significance" based upon the variant pathogenicity guidelines from the American College of Medical Genetics and Genomics (ACMG). This paper highlights the genetic diversity of vision disorders in the Pakistani population and reports the identification of four novel mutations in families who segregate clinically heterogeneous eye diseases. Our results give insight into the genotype-phenotype correlations of AIPL1, FOXE3, MERTK, and GUCY2D variants.

Genetic transformation of Sr22 gene in a high yielding susceptible cultivar of commercial wheat (Triticum aestivum L.).

In this study, the Sr22 gene was isolated and prepared for transformation in disease-susceptible commercial high-yielding wheat (Triticum aestivum L.) cultivar Lasani-2008. The Sr22 fragment was initially inserted in plasmid pUC57 for sequence confirmation before performing further experiments. After confirmation, Sr22 was subcloned in pGreen0029 which helped in further cloning and ligation. pUC57-Sr22 was restricted with Nru1 and BamH1, while pGreen0029 was restricted with EcoRV and BamH1 and ligated. From pGreen0029, Sr22 was eluted and ligated in pJIT163 to insert the 2 × 35S promoter and CaMV terminator using Xho1 and BamH1 and Sal1. At this stage, the expression cassette was completed. The 2 × 35Sp-Sr22-CaMVt was then ligated in pGreen0029 and transferred to Agrobacterium along with pSOUP. pSOUP helped pGreen0029 to insert 2X35Sp-Sr22-CaMVt in the callus of Lasani-2008, along with kanamycin-resistant gene. Transgenic callus was used for regeneration of the whole plant by tissue culture. Transgenic plants were further tested by PCR, qPCR and SDS-PAGE. The transgenic Lasani-2008 showed substantial resistance against stem rust in both seedling and adult plant stages. The results also showed that transgenic Lasani-2008 has increased average yield of grains (i.e., 4893 ± 148 kg/ha) as compared to non-transgenic Lasani-2008 (i.e., with average yield of gains 4762 ± 103 kg/ha). Sr22 containing lines and the transgenic developed in this study can be used in breeding systems. Transgenic seeds developed will be shared with breeding institutes and breeders should use this information to develop new varieties.

Characterization of antifungal metabolites produced by Lactobacillus plantarum and Lactobacillus coryniformis isolated from rice rinsed water.

A recent spike in demand for chemical preservative free food has derived the scientific community to develop natural ways of food preservation. Therefore, bio-preservation could be considered as the great alternative over chemical ones owing to its potential to increase shelf-life and nutritional values of foodstuffs. In the present study, lactic acid producing bacterial species were isolated from rice rinsed water and identified by 16S rRNA gene sequencing as Lactobacillus plantarum BCH-1 (KX388380) and Lactobacillus coryniformis BCH-4 (KX388387). Antifungal metabolites from both Lactobacillus species were extracted by polarity-based solvents in which ethyl acetate showed remarkable antifungal activity against Aspergillus flavus and Aspergillus fumigatus by disc diffusion assay. Different organic acids and fatty acids have been identified by reversed-phase high-performance liquid chromatography (RP-HPLC) and gas chromatography-mass spectrometry (GC-MS) analysis, respectively. Lactic acid and citric acid were the major organic acids found in ethyl acetate fractions of L. plantarum and L. coryniformis, respectively. Similarly, 9,12-otadecadienoic acid (Z,Z)-methyl ester and hexadecanoic acid, methyl ester were the major fatty acids found in n-hexane fractions of L. plantarum and L. coryniformis respectively. Moreover, the isolation of novel antifungal metabolites from locally isolated Lactobacillus species was focused and it was revealed that organic acids are important contributors towards antifungal potential. A novel fatty acid (i.e. 12-hydroxydodecanoic acid) has also been explored and found as potential metabolite against filamentous fungi. Conclusively, various metabolites isolated from non-dairy source showed antifungal activity especially against Aspergillus species. Hence, these metabolites have been considered as a good choice for bio-preservation.

Exogenously Applied Ascorbic Acid-Mediated Changes in Osmoprotection and Oxidative Defense System Enhanced Water Stress Tolerance in Different Cultivars of Safflower (Carthamus tinctorious L.).

The present study was conducted to examine the effect of exogenously applied ascorbic acid (AsA) on osmoprotectants and the oxidative defense system in four cultivars (16171, 16183, 16207 and 16246) of safflower under well-watered and water deficit conditions. Water stress (60% field capacity) significantly decreased the shoot and root fresh and dry weights, shoot and root lengths and chlorophyll contents in all four safflower cultivars, while it increased the leaf free proline, total phenolics, total soluble proteins, hydrogen peroxide content and activities of catalase, superoxide dismutase and peroxidase enzymes. Foliar-applied (100 mg L-1 and 150 mg L-1) ascorbic acid caused a marked improvement in shoot and root fresh and dry weights, plant height, chlorophyll and AsA contents as well as the activity of peroxidase (POD) enzyme particularly under water deficit conditions. It also increased the accumulation of leaf proline, total phenolics, total soluble proteins and glycine betaine (GB) content in all four cultivars. Exogenously applied AsA lowered the contents of MDA and H2O2, and the activities of CAT and SOD enzymes. Overall, exogenously applied AsA had a positive effect on the growth of safflower plants under water deficit conditions which could be related to AsA-induced enhanced osmoprotection and regulation of antioxidant defense system.

Enhancement of nutraceutical and antioxidant potential of sunflower hybrid seed varieties through chemical priming.

Pathogenic agents cause an increased risk of various fatal diseases and there is a need to reduce this risk using medicinal plants and their seeds. The present research work was designed to study the efficacy of different sunflower seed hybrid varieties (i.e. FH622, FH620, FH615, FH613 and FH545) chemically primed with potassium nitrate as natural antioxidant and antimicrobial agent. Antioxidant potential was determined using DPPH test, reducing power, TPC and TFC. Antibacterial activity was determined against Gram positive and Gram negative bacterial species. After one week, the germination data including mean germination and percentage of final emergence was calculated. It was found that seed varieties FH620 and FH615 have higher values of mean germination as compared to FH545 while FH615 has higher percentage of final emergence as compared to FH620 and FH545. High phenolic and flavonoid contents were observed in FH620 and FH615 as compared to FH545 variety. It was also observed that seed variety FH615 when treated with KNO3 solution had significantly high germination as well as antioxidant parameters and antibacterial activity as compared to other varieties. Similarly FH615 showed high antibacterial activities against Gram positive Staphylococcus aureus and Gram-negative Escherichia coli bacteria. This study showed that all selected sunflower hybrids have good antioxidant and antibacterial potentials that would further be used for different trials to cure different pathogen related diseases, and these are natural sources of antioxidants for commercial and therapeutic applications.

Secreted frizzled-related protein 4 and its implication in obesity and type-2 diabetes.

Secreted frizzled-related protein 4 (SFRP4) is a member of secreted protein family with sequence similarity to frizzled receptors of wingless-related integration site (Wnt) signaling pathways. These proteins control diverse functions from embryonic development to adults in many organisms including humans. Initially, SFRPs were recognized as antagonists of Wnt signaling and supposed to interact with Wnts. Further research demonstrated their interactions to frizzled receptors and a functional diversity was related to these proteins, Wnt signaling potentiation in addition to modulation. SFRP4 is the largest member of SFRP family and is implicated in many diseases including obesity, type 2 diabetes (T2D), and cancer. SFRP4 acts as a biomarker for T2D and was expressed several years before clinical diagnosis of disease. This review mainly focusses on the role of SFRP4 in obesity and how it can lead to ß-cell failure and ultimately to T2D. The role of SFRP4 in adipose tissues causing increased production of adipokines lead to the oxidative stress in pancreas that particularly have low amount of antioxidant enzymes in pancreatic ß-cells leading to failure in exocytosis of insulin containing granules causing T2D. Obesity-induced inflammation is a principal factor in pathogenesis of insulin resistance as well as metabolic syndrome. Pro-inflammatory cytokines have potential to cause insulin resistance in skeletal muscles, adipose tissue, and liver via inhibition of insulin signal transduction. Secretion of SFRP4 is mediated by interleukin 1-ß (IL1-ß). This review highlights the molecular mechanisms by which SFRP4 leads to T2D. Understanding of molecular mechanism and targeting SFRP4 could help to eradicate or reduce chances of developing T2D.

Magneto-Priming Improved Nutraceutical Potential and Antimicrobial Activity of Momordica charantia L. Without Affecting Nutritive Value.

The need for some economic strategies for increased growth and nutraceuticals of medicinal plants is well acknowledged now. It was hypothesized that external magnetic field treatment (MFT) of seeds affecting internal magnet of cells may affect growth and metabolism. In this study, seeds were subjected to pre-sowing magnetic field (50 mT at 5 mm for 5 s). At vegetative stage, the leaf growth, chlorophyll content, catalase (CAT), peroxidase (POD), amino acids, proteins, flavonoids, soluble sugars, total soluble phenolics, carotenoids, anthocyanins, phenolic profile (HPLC based), and antimicrobial activity of leaves (in terms of the minimum inhibitory concentration against Staphylococcus aureus and Pseudomonas aeruginosa) were studied. Yield was evaluated for nutritive components in fruit (peel+pulp) and peel. MFT improved germination percentage, growth, leaf chlorophyll, antimicrobial activity, peel amino acids, phenolics, and POD with negligible effect on fruit nutritive value. Moreover, photosynthetic pigments and cinnamic acid exhibited direct correlation with antimicrobial potential against both pathogens. However, sinapic acid showed positive correlation against Staphylococcus aureus only. Cinnamic acid, coumaric acid, syringic acid, and quercetin were in direct correlation against Pseudomonas aeruginosa; it was directly correlated with total flavonoids too. In conclusion, magnetic field can be used to manipulate plant cell metabolism promising improvement of growth, antimicrobial activity, and phenolics of interest.

Purification and modification of Cordia myxa gum to enhance its nutraceutical attribute as binding agent.

The aim of the present study was to purify, hydrolyze and modify the Cordia myxa gum to document its binder potential in pharmaceutical tablets formulation. The hydrolysis and modification was carried out to remove impurities, roughness, increase thermal stability and to improve the functional properties of biopolymers. Physiochemical properties such as pH, solubility, viscosity, swelling index, bulk and tapped density was performed prior to investigate binder potential. The binder potential of Cordia myxa gum was studied in its different forms such as crude, purified, modified and hydrolyzed in paracetamol tablets and was compared with standard hydroxypropyl methylcellulose (HPMC) being used as synthetic binder. Tablets were prepared by direct compression method and evaluated for weight uniformity, hardness, friability, disintegration time and dissolution analysis. Prepared tablets with selected gums exhibit faster and slower dissolution profile in the same dissolution system. The crude gum has high dissolution rate whereas the hydrolyzed and modified gums showed less dissolution rate. The hydrolyzed and modified gums having faster release rate and it could be helpful in conventional tablet formulations efficiently as compared to synthetic HPMC binder.

Phenolic acid profiling and antiglycation studies of leaf and fruit extracts of tyrosine primed Momordica charantia seeds for possible treatment of diabetes mellitus.

The increasing risk of variety of fatal diseases including diabetes mellitus is imposing serious challenge to chemist, biologists and clinicians. Due to the side effects of the chemotherapy, worldwide it is thinking that phyto-medicine are more effective to cope continuously increasing risk of fatal diseases without any side effect. Seed priming is a strategic pre-sowing semi-bioengineering technique which has ability to improve the growth rate and biologically active compounds in short time. Among seed priming techniques, tyrosine seed priming most frequently used because amino acids provide best growth media for nutritional food crops. Seeds of Momordica charantia were subjected to the pre-sowing tyrosine solution. Different growth parameters including growth emergence rate, seedling vigor, growth and weight of root, shoot and leaf were studied. The results showed positive effect on Momordica charantia seed growth and phenolic acids production i.e. ferulic acid - 43.95 ppm and sinapic acid - 18.39 ppm. The antiglycation assay showed 23.45±1.23% antiglycation activity of primed-seed fruit extract as compare to control seed fruit extract (0.87±0.03%). On the basis of the results, it is concluded that tyrosine primed seed fruit extract could effectively be further tested for pre-clinical and clinical studies to manage diabetes mellitus disease.

Zika Virus: A Critical Analysis and Pharmaceutical Perspectives.

Zika virus belongs to family of viruses 'Flaviviridae' and spreads mostly by daytime-active Aedes mosquitoes. The resulting infection is known as Zika fever. It is usually asymptomatic or often causes mild symptoms, which are very similar to dengue fever. The Aedes aegypti mosquito is also responsible for dengue and chikungunya viruses. Zika virus can spread by crossing the placental barrier from a pregnant mother to a fetus, which can result in microcephaly, severe brain malformations including Guillain-Barre syndrome (GBS), and other birth defects. Until now, there is no specific treatment of Zika fever disease, and Zika virus illness cannot be prevented by medications or vaccines. According to WHO, no vaccine is likely to be available until 2020. The only way of preventing this disease is to prevent the mosquito bites. This article presents the history of Zika virus, its reported cases including microcephaly and GBS, and a comparison of its symptoms with those of dengue and chikungunya diseases, as well as preventive measures. With advances in research and technology, knowledge about the Zika virus has grown, yet some questions remain unanswered regarding Zika virus's genetic diversity, pathophysiology, transmission vectors and reservoirs, potential synergetic of coinfection with other related arboviruses, and treatment. These problems highlight the need for further research to achieve adequate the surveillance, infection management, optimized treatment, and public health mediations in Zika virus outbreaks. This article contributes to our understanding of the disease mechanism, genome structure, diagnosis, transmission, and preventive strategies to combat Zika virus infection.

Post interferon therapy decreases male fertility through gonadotoxic effect.

Prevalence of hepatitis C virus (HCV) has been seen in more than 15% of Pakistani population. For the treatment of this infection, only two medicines, interferon, and ribavirin were approved in 1998. The concerned physicians evaluate side effects of these two antiviral drugs only during the treatment period. The long-term extra hepatic side effects are being neglected. This retrospective study was conducted with reference to induced infertility in HCV treated 40 male patients from the period 2008-2015. Possible effects of interferon therapy on fertility hormones and seminal parameters were assessed. Level of fertility hormones like serum Follicle Stimulating Hormone (FSH), Luteinizing Hormone (LH), and testosterone was measured. For seminal parameters, guidelines from World Health Organization (WHO) were followed. Among forty cases of HCV patients who received interferon, only 14 (35%) have children and 26 (65%) could not conceive (p = 0.0372). After HCV treatment, HCV positive patients showed a significant change in the level of FSH, LH (p<0.05). Especially, it decreased testosterone level (p=0.0096). Similarly, HCV treatment significantly decreased sperm count (p=0.001) and motility (p=0.0005).

A novel mutation in RDH5 gene causes retinitis pigmentosa in consanguineous Pakistani family.

Retinitis pigmentosa (RP) is the most frequent genetically and clinically heterogeneous inherited retinal degeneration. To date, more than 80 genes have been identified that cause autosomal dominant, autosomal recessive and X linked RP. However, locus and allelic heterogeneity of RP has not been fully captured yet. This heterogeneity and lack of an accurate genotype phenotype correlation makes molecular dissection of the disease more difficult. The present study was designed to characterize the underlying pathogenic variants of RP in Pakistan. For this purpose, a large consanguineous family with RP phenotype showing autosomal recessive mode of inheritance was selected after a complete ophthalmological examination. Next generation sequencing was used for the identification of molecular determinant followed by Sanger-sequencing for confirmation. After sequence analysis a novel homozygous missense mutation, (c.602 C > T) in exon 4 of the RDH5 gene (MIM: 601617) was identified. This mutation resulted in substitution of phenyl alanine for serine at amino acid 201 (p.Ser201Phe) of the RDH5 gene. The same mutation was not detected in the 200 ethnically-matched control samples by Sanger sequencing. The identified mutant allele segregated in homozygous fashion in all the affected individuals of pedigree. Identification of this mutation reveals the allelic heterogeneity of RDH5 in patients with RP phenotype. The findings of this study demonstrate the clinical significance of next generation sequencing to understand the molecular basis of diseases and would help to reveal new proteins and their function in visual cycle will pave the way for early diagnosis, genetic counseling and better therapeutic inventions.