Novel Noninvasive Serum Biomarkers for Prompt Diagnosis of Breast Carcinoma.

Immune cell infiltration is associated with improved prognosis in the microenvironment of breast cancer. The incidence of breast cancer in Pakistan is 2.5 times higher than that in neighboring countries of Asia, accounting for 34.6% of female cancers. The objectives of this study were to compare and determine apoptotic mediators and biomarkers for breast carcinoma, such as serum granzyme B, cytochrome C, and vitamin D by ELIZA and calcium spectrophotometrically. Study groups were differentiated into malignant breast disease G-I, benign proliferative breast disease G-II, and healthy control group G-III. The immune-related prognostic markers and therapeutic targets were determined through the interaction of proteins by molecular docking and AutoDock Vina software. The level of granzyme B and cyt C was higher in Group-I, -II, and -III. Likewise, the mean vitamin D level was greater in Group-I than those in other groups. Through SwissDock, the proteins granzyme B and cyt C with vitamin D, single amino acid residue MET34 (H-bond 2.75 Å), and ILE81(H-bond 2.092 Å) were revealed to actively participate in interactions. This study reveals granzyme B and cyt C as biomarkers for malignant breast disease and benign proliferative breast disease, while hypovitaminosis D and hypocalcemia are complications or comorbidities of breast cancer.

Anti-arthritic and toxicological evaluation of ethanolic extract of Alternanthera bettzickiana in rats.

In many developing countries, medicinal plants have long been used for therapeutic purposes due to their low cost and toxicity. This study evaluated the safety and anti-arthritic potential of Alternanthera bettzickiana ethanolic extract (ABEE). Acute oral toxicity (OECD 425) was tested in the safety evaluation. A limit test was used to identify the LD50 value. For an acute oral toxicity study a dose of 2000 mg/kg of ABEE was given orally to the treatment group, and the control group received distilled water at a rate of 10 ml/kg. Biochemical, hematological, and histopathological analyses were performed after 14 days. A formaldehyde 2% w/v solution was injected via i.p. to rats of all groups to prepare the arthritic model. Five groups were divided into control (D.H2O), standard (Diclofenac), and three groups receiving the plant extract at dose levels of 125 mg/kg, 250 mg/kg, and 500 mg/kg respectively. Treatment was continued for 10 days. Paw diameter and hematological and biochemical variables were quantified. ELISA was performed for the estimation of inflammatory cytokines. In the acute oral toxicity study, no mortality or morbidity were observed, so the LD50 of this plant was greater than 2000 mg/kg. ABEE decreased the paw diameter with the restoration of hematological and biochemical changes. SOD and CAT levels were increased while decreasing the MDA, NO, TNF-α, and IL-6 levels in arthritic rats. It is concluded that the use of A. bettzickiana has low toxicity, and it can be used for the treatment of arthritis.

Proteome based mapping and reverse vaccinology techniques to contrive multi-epitope based subunit vaccine (MEBSV) against Streptococcus pyogenes.

Streptococcus pyogenes is a root cause of human infection like pharyngitis, tonsillitis, scarlet fever, impetigo, and respiratory tract infections. About 11 million individuals in the US suffer from pharyngitis every year. Unfortunately, no vaccine against S. pyogenes is available yet. The purpose of this study is to create a multiepitope-based subunit vaccine (MEBSV) targeting S. pyogenes top four highly antigenic proteins by using a combination of immunological techniques and molecular docking to tackle term group A streptococcal (GAS) infections. T-cell (HTL & CTL), B-cell, and IFN-γ of target proteins were forecasted and epitopes having high antigenic properties being selected for subsequent research. For designing of final vaccine, 5LBL, 9CTL, and 4HTL epitopes were joined by the KK, AAY, and GPGPG linkers. To enhance the immune response, the N-end of the vaccine was linked by adjuvant (Cholera enterotoxin subunit B) with a linker named EAAAK. With the addition of adjuvants and linkers, the construct size was 421 amino acids. IFN-γ and B-cell epitopes illustrate that the modeled construct is optimized for cell-mediated immune or humoral responses. The developed MEBSV structure was assessed to be highly antigenic, non-toxic, and non-allergenic. Moreover, disulphide engineering further enhanced the stability of the final vaccine protein. Molecular docking of the MEBSV with toll-like receptor 4 (TLR4) was conducted to check the vaccine's compatibility with the receptor. Besides, in-silico cloning has been carried out for credibility validation and proper expression of vaccine construct. These findings suggested that the multi-epitope vaccine produced might be a potential immunogenic against Group A streptococcus infections but further experimental testing is required to validate this study.

SNP of HMGCR and Apo E genes and their impact in response to statin therapy in hypercholesterolemic and hypertriglyceridemic patients in Pakistan.

Coronary artery disease (CAD) and the problems associated with it are the most prominent causes of death in the whole world. Statins are accustomed to lower lipid levels in CAD patients. The target of this study was to analyze whether or not common variations in HMGCoA Reductase (HMGCR) and Apolipoprotein E (ApoE) genes are responsible for metabolism of lipid and statin that modify the impact of statins on serum level of lipids and lipoprotein concentrations in Coronary heart disease patients. One hundred CAD patients were registered for the study. At the start of the study biochemical measurements were performed to work out the baseline levels. Patients were treated with twenty mg Lipitor for one month and biochemical measurements were tested again. According to the post-treatment, LDL-c levels, patients were divided into a pair of group as non-responders and responders, independently. The information concerning the risk factors like smoking, alcohol consumption etc. was conjointly obtained. DNA was extracted from peripheral blood. The presence of rs17244841 and rs17238540 mutations in HMGCR and έ2, έ3 and έ4 variants of ApoE were settled by performing RT-PCR. Results were assessed statistically. HMGCR mutations were principally found in responders and ε4 variant of ApoE was principally found in non-responders. It was found that the presence of HMGCR mutations causes a big reduction in total cholesterol and LDL-c levels. Conjointly, the presence of έ2 variant of Apo E causes a statistically vital increase in triglyceride levels. Our findings should be investigated by different researchers to clarify the mechanism.

The prospects of antimicrobial coated medical implants.

The implants are increasingly being a part of modern medicine in various surgical procedures for functional or cosmetic purposes. The progressive use of implants is associated with increased infectious complications and prevention of such infections always remains precedence in the clinical settings. The preventive approaches include the systemic administration of antimicrobial agents before and after the surgical procedures as well as the local application of antibiotics. The relevant literature and existing clinical practices have highlighted the role of antimicrobial coating approaches in the prevention of implants associated infections, although the applications of these strategies are not yet standardized, and the clinical efficacy is not much clear. The adequate data from the randomized control trials is challenging because of the unavailability of a large sample size although it is compulsory in this context to assess the clinical efficacy of preemptive practices. This review compares the efficacy of preventive approaches and the prospects of antimicrobial-coated implants in preventing implant-related infections.

Platelet storage pool disorder: multidisciplinary planning in pregnancy.

A 32-year old primigravida woman presented for antenatal care giving a history that her mother had platelet storage pool disorder (PSPD). The patient was subsequently diagnosed with PSPD during her pregnancy and had a caesarean delivery for breech presentation at 39 weeks. In this paper, we discuss the basic science, inheritance pattern, symptoms and management of this condition, alongside the antenatal and intrapartum and postnatal management specific to it, highlighting the need for a multidisciplinary approach to care. PSPD refers to a group of rare conditions involving defects in platelet granule storage or secretion, which leads to abnormal aggregation and activation of platelets. There are both genetic and acquired forms of the condition. It is a functional platelet disorder, meaning platelet counts will usually remain in the normal range. The diagnosis may be suspected due to characteristic signs and symptoms, but patients may also be asymptomatic. There have been only a few documented cases of pregnant women with PSPD; therefore, management is not clear. Vaginal delivery is not contraindicated, however, postpartum haemorrhage should be anticipated and planned for the use of deamino D-arginine vasopressin (DDAVP), tranexamic acid, prophylactic oxytocics and prompt access to blood products, including platelets, if required. This case highlights the need for effective multidisciplinary teamwork between obstetricians, anaesthetists and haematologists to ensure high-quality care and enable careful intrapartum management planning.

Formulation and Evaluation of a Clove Oil-Encapsulated Nanofiber Formulation for Effective Wound-Healing.

Wound-healing is complicated process that is affected by many factors, especially bacterial infiltration at the site and not only the need for the regeneration of damaged tissues but also the requirement for antibacterial, anti-inflammatory, and analgesic activity at the injured site. The objective of the present study was to develop and evaluate the natural essential oil-containing nanofiber (NF) mat with enhanced antibacterial activity, regenerative, non-cytotoxic, and wound-healing potential. Clove essential oil (CEO) encapsulated in chitosan and poly-ethylene oxide (PEO) polymers to form NFs and their morphology was analyzed using scanning electron microscopy (SEM) that confirmed the finest NFs prepared with a diameter of 154 ± 35 nm. The successful incorporation of CEO was characterized by Fourier transform infra-red spectroscopy (FTIR) and X-ray diffractometry (XRD). The 87.6 ± 13.1% encapsulation efficiency and 8.9 ± 0.98% loading of CEO was observed. A total of 79% release of CEO was observed in acidic pH 5.5 with 117% high degree of swelling. The prepared NF mat showed good antibacterial activity against Staphylococcus aureus and Escherichia coli and non-cytotoxic behavior against human fibroblast cell lines and showed good wound-healing potential.

Efficacy of Probiotics Supplementation on Growth Performance, Carcass Composition and Hematological Parameters of Cyprinus carpio Fingerlings Fed Corn Gluten Meal-Based Diet

Abstract Plant based protein sources are one of the best, cost effective and easily available protein sources being used in fish feed. But due to a lower number of micro-biota in fish gut plant meal based diets cannot be digested and absorbed well in fish body. Probiotics were supplemented at 0, 1, 2, 3, 4 and 5 gkg-1 levels in fish feed for formulating one control and five test diets. In this study, three replicates of each treatment were used and number of fingerlings was 15 in each replicate. The C. carpio (common carp) fingerlings were fed at 5% of live wet weight on their prescribed diet twice daily. The results revealed that supplementation of probiotics in corn gluten meal based diets significantly (p<0.05) improved growth performance, carcass composition and hematological parameters. Most optimum values of growth performance parameters were noted at 2 gkg-1 level of probiotics supplemented diet. C. carpio fingerlings fed corn gluten meal based diet supplemented with 2 gkg-1 level of probiotics indicated significant (p<0.05) improvements in crude protein (17g) crude fat (9g) and gross energy (3 kcalg-1) whereas higher red blood cells (RBCs), white blood cells (WBCs) and hemoglobin (Hb) was also recorded in fish blood when fed 2 gkg-1 probiotics level diet. From these results, it was concluded that 2 gkg-1 probiotics supplementation in corn gluten meal based diet is optimum for improving growth performance, body composition and hematology of C. carpio fingerlings.

In vitro Characterization and Release Studies of Combined Nonionic Surfactant-Based Vesicles for the Prolonged Delivery of an Immunosuppressant Model Drug.

BACKGROUND: Cyclosporine A (CsA) is an exceptional immunosuppressant used for the treatment of immune disorders. Niosomal vesicles are promising drug carriers that are formed by self-association of nonionic surfactants and cholesterol in an aqueous phase. The objective of the study was to formulate combined nonionic surfactant based vesicles and to evaluate their in vitro characterization, release studies and in vivo studies. MATERIALS AND METHODS: Five niosomal formulations (F7 to F11) were prepared using the thin film hydration method. The molar ratio of cholesterol and non-ionic surfactant taken was 1:1. In formulation F10, the combination of surfactants Span 20 and Brij 35 was used. The niosomes were characterized by zeta sizer and SEM for particle size analysis, in vitro drug release and stability studies. The pharmacokinetic studies were conducted on healthy albino rabbits. RESULTS: The size of niosome was found in the range of 427.1 nm to 972.3 nm. SEM image of optimized formulations F10 exhibit the spherical nature of niosomal vesicles. DSC thermograms of niosomal formulations exhibited a broadened endothermic peak. The stability study exhibited that all formulations are stable and negligible change of vesicle size and entrapment was observed with time. The percentage drug release was significantly higher as compared to CsA plain dispersion for all niosomal formulations at pH 1.2 and 7.4. The release kinetic behavior showed that all preparations were best described by zero order and can release active ingredient in a sustained manner. The pharmacokinetic data showed the test formulation (F10) possessed greater bioavailability as compared to the reference formulation (CsA aqueous dispersion). CONCLUSION: The formulation F10 demonstrated a comparatively more delayed rate of release with enhanced dissolution as compared to a single surfactant scheme. The F10 formulation can be a remarkable nanotechnology for prolonged delivery of CsA orally with improved dissolution profile and bioavailability.

Chemical Composition and Bioactivities of Essential Oil from Leaves of Syzygium cumini (L.) Skeels Native to Punjab, Pakistan.

Herbal medicines are widely used for the treatment of different types of diseases like skin and throat infections and other diseases in developing countries. Syzygium cumini (L.) Skeels fruit, leaves and bark were used for the remedies of different diseases anciently. The aim of the present study was to evaluate the chemical profile of Syzygium cumini leaves essential oil (EO) from Punjab, Pakistan. The essential oil was isolated using hydrodistillation technique and analyzed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC/MS). Free radical scavenging capacity and antioxidant activity were assessed by using DPPH radical scavenging ability, inhibition of linoleic acid peroxidation, bleaching of ß-carotene in linoleic acid system and reducing power assays. Antimicrobial potential was assessed by disc diffusion assay and measurement of minimum inhibitory concentration (MIC) using resazurin microtiter-plate assay. The anti-heme biocrystallization activity of EO was also assessed. The major components (>3%) found in Syzygium cumini leaves EO were ß-farnesene (3.42 %), caryophyllenol (3.46 %), terpinen-4-ol (3.61 %), ß-myrcene (3.90 %), γ-cadinene (4.09 %), fenchol (4.22 %), cis-ß-ocimene (4.40 %) and 5-methyl-1,3,6-heptatriene (4.90 %). Excellent antioxidant, antimicrobial and weak antimalarial potential was observed. It can be concluded that Syzygium cumini leaves EO has potential application for food and pharmaceutical industries.

Development of 99mTc-SDP-choline SPECT radiopharmaceutical for imaging of cerebrovascular diseases.

Cerebrovascular diseases are known as serious public health problem worldwide, which can be addressed more precisely through molecular imaging of non-functional brain cells. CDP-choline is an active cerebrovascular chemotherapeutic agent that can be used for diagnosis of cerebrovascular diseases post radiolabeling with γ-emitter radioisotopes. In this study we developed 99mTc labeled CDP-choline for imaging of cerebrovascular diseases particularly alzheimer, stroke, and parkinson's diseases. The radiosynthesis reaction resulted 97.47±2.34% radiochemical with promising stability, that is, >95% up to 6 h in blood serum. The biodistribution study in healthy mice revealed non-accumulated uptake of radiochemical in key body organs; in brain it was 8.59±1.11% ID/g at 1h post-injection which washed-out leaving behind 0.87±0.61% ID/g at 24 h post-injection. The over-all data revealed the 99mTc-CDP-choline could be a good candidate for further imaging investigations in diseased animal model.

Binary inclusion complexes of diflunisal with ß-cyclodextrin and hydroxypropyl-ß-cyclodextrin: Preparation and characterization.

Low aqueous solubility and bioavailability is the limiting factor to achieve desired therapeutic efficacy for variety of new and existing drug moieties. The goal of the present study was to explore the effects of ß-cyclodextrin (ßCD) and hydroxypropyl-ß-cyclodextrin (HPßCD) on the solubility and dissolution profile of diflunisal (DIF) prepared by using two different methods (physical mixing and solvent evaporation) at DIF-cyclodextrins weight ratios of 1:1, 1:2 and 1:4. The phase solubility studies demonstrated that DIF solubility increased proportionally with an increase in ßCD and HPßCD concentration. The inclusion complexes were subjected to characterization of scanning electron microscopy (SEM), fourier transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC) and X-ray diffractometry (XRD). Solvent evaporation yielded higher DIF solubility than physical mixing method. HPßCD-DIF inclusion complexes yielded higher dissolution profile than ßCD complexes when prepared under same experimental design. FTIR, DSC and XRD confirmed the successful inclusion of DIF into cyclodextrin (ßCD/HPßCD) by both preparation methods with enhanced water solubility and drug release in comparison with pure drug.

Computational screening of phytochemicals against survivin protein: A potent target for cancer.

Survivin (IAP proteins) is considered as a significant target for anticancer drug research owing to its upregulation in tumor cells to mediate resistance to apoptotic stimulus. The current study aimed to investigate phytochemicals as inhibitors of survivin with caspases to reactivate the functioning of caspases through molecular docking. The compounds namely 2(R), 4(R)-dihydroxypyrrolidine, 4-hydroxy-2-(4-methoxyphenyl)-1,1-dioxo-3,4-dihydrothieno[3,2-e]thiazine-6-sulfonamide, 2,3-Diketo-L-gulonic acid, (3-hydroxy-2-octadeca-9,12-dienoyloxypropyl) octadecanoate, 2-[[4-[[4-[(4-formamido-1-methylimidazole-2-carbonyl)amino]-1-methylimidazole-2-carbonyl]amino]-1-methylimidazole-2-carbonyl]amino]ethyl-dimethylazanium, Picolinic acid and (2-Hydroxy-5-nitrophenyl) dihydrogen phosphate successfully bind inside the pocket of survivin. ADMETsar was used to evaluate the anticancer potential of selected compounds. These compounds can be proposed as effective inhibitors, disrupting the survivin-caspases interaction and reactivating the caspases function of apoptosis. The study might facilitate the development of cost-effective and natural drugs against cancer. However, further validation is essential for confirmation of its drug efficacy and bio-compatibility.

Effects of chelating agents on heavy metals in Hepatitis C Virus (HCV) patients.

Heavy metals are released into the environment through both human and natural sources, may have a direct hepatic toxicity and are involved in chronic liver diseases. Modification in the regulation of heavy metals metabolism enhanced hepatitis c virus (HCV) replication which ultimately reduced outcomes of anti-viral therapy in chronic HCV patients. Chelation therapy with new drugs seems to eradicate HCV and may prevent liver complications. The present study was planned to explore the effects of MiADMSA (lipophilic chelating agent) for achieving maximum heavy metals elimination in hepatitis c virus patients with minimum side effects. For this purpose concentration of heavy metal was determined in HCV patients and established correlation of heavy metals between healthy persons and HCV patients. Atomic absorption spectrophotometer (AAS) was used to explore them. Concentrations of heavy metal in different samples (blood serum, nails and hair) of patients and healthy individuals. Result revealed that heavy metals (Lead, Cobalt, Cadmium, Manganese, Iron and Cooper) concentration were significantly higher in blood of HCV patients as compared to normal persons, but some metals like Ni and Zn were present in normal concentration and in low concentration respectively. After chelation with monoisoamyl DMSA (MiADMSA) a significant amount of heavy metals was excreted in the urine in a dose dependent manner. It was generally observed from the results that TDS is a better treatment option than BD for chelation of heavy metals in hepatitis c virus patients. This chelation therapy will be helpful to reverse the HCV related health problems.

Analysis of toll-like receptors-9 (TLR9) gene polymorphism (rs5743836) in Pakistani patients with HCV.

Toll-like receptors (TLRs) are innate immune receptors that mediate the inflammatory response during HCV infections. The goal of this study was to evaluate the association of TLR9 gene polymorphism (rs5743836) in Pakistani patients infected with genotype 3a of HCV. Total 500 subjects were recruited, 400 HCV patients and 100 healthy individuals. Genotyping of TLR9 (-1237T/C, rs5743836) was carried out in 400 HCV patients (323 interferon responders and 77 interferon non responder) and control group by applying High resolution melting (HRM) curve assay. No remarkable differences in distribution of genotype between HCV (p<0.0001; OR= 3.21, 95% CI= (2.514.12) and control groups (p<0.0001; OR=0.092, 95%CI= (0.0580.14) were observed. In conclusion TLR9-1237T/C gene polymorphism may not be considered as a molecular risk for patients with HCV in Pakistan.

Phytochemical analysis and hepatoprotective effect of polyherbal formulation on CCl4 induced hepatotoxicity in mice.

The potent phytotherapeutic modalities against the hepatotoxicity have motivated us to explore numerous plants and polyherbal preparations because conventional drug discovery is more expensive and tedious. So, this study was conducted to evaluate the hepatoprotective potential of a polyherbal formulation (PHF), comprising of Solanum nigrum, Silybum marianum, Atrmesia absinthium, Achillea millifolium and Cichorium intybus against carbon tetrachloride(CCl4) induced hepatotoxicity in experimental rats. CCl4intoxicationinduced vacuole formation and fastdegeneration so selective liver enzymes including alanine aminotransferase (ALT) and aspartate aminotransferase (AST), alkalinephosphatase (ALP) and total bilirubin in rat's plasma,as well as liver histological architecture, were used to evaluate the effect of herbal treatments with different doses (ranging 100-500 mg/kg) for two weeks. Statistical analysis showed that PHF significantly (P<.05) improved the level of liver enzymes as well as improve the liver architecture comparative to control groups. It could be concluded from current findings that PHF prepared from Solanum nigrum, Silybum marianum, Atrmesia absinthium, Achillea millifiloium and Cichorium intybus have some hepatoprotective activities.

Effect of caffeine on anti-clotting activity of warfarin in healthy male albino rabbits.

Drug-drug interactions are most commonly occurring phenomenon in clinical practice. Many physicians are afraid of being involved in an allegation of malpractices due to the occurrence of any severe interaction. These interactions not only occur between drugs but also between any kind of food, tobacco smoke, caffeine and alcohol etc. Therefore, the present study was directed to inspect the effect of caffeine on the anticoagulation activity of warfarin in healthy adult male albino rabbits. Blank blood samples were collected from each rabbit. Rabbits were given warfarin (0.5mg kg-1) orally via stomach tube and blood samples were collected in PT/INR vials at various intervals. After a washout period of 14 days, warfarin was orally administrated at same dose rate along with caffeine (5 mg kg-1 every twelve hours for three days) and same sampling schedule was repeated. Prothrombin time (PT) and the international normalized ratio (INR) of blood samples were determined to estimate changes in the anticoagulation activity of warfarin after its concurrent administration with caffeine. The PT data revealed that Rmax and AUC increased significantly (P<0.05) from 1991.6 and 60.5 to 2124.8 and 67.5, respectively, before and after co-administration. Similarly, a significant (P<0.05) increase was observed in Rmax and AUC of INR from 6.42 and 153.7 to 7.4 and 167.5, respectively, alone and along with caffeine. However, no change was observed in Tmax associated with PT and INR either the drug was administered alone or in combination with caffeine. It was concluded that caffeine has the capacity to inhibit the metabolism of warfarin and enhance its plasma concentration and hence anticoagulant effects. Thus, patients should be advised to limit the frequent use of caffeine-rich products i.e. tea and coffee during warfarin therapy.

Acidic and enzymatic saccharification of waste agricultural biomass for biotechnological production of xylitol.

BACKGROUND: The plant biomass and agro-industrial wastes show great potential for their use as attractive low cost substrates in biotechnological processes. Wheat straw and corn cob as hemicellulosic substrates were acid hydrolyzed and enzymatically saccharified for high xylose production. The hydrolysate was concentrated and fermented by using Saccharomyces cerevisiae and Kluyveromyces for production of xylitol. RESULTS: Acid hydrolysis of wheat straw and corn cob in combination with enzymatic hydrolysis showed great potential for production of free sugars from these substrates. Kluyveromyces produced maximum xylitol from acid treated wheat straw residues with enzymatic saccharification. The percentage xylitol yield was 89.807 g/L and volumetric productivity of 0.019 g/L/h. Kluyveromyces also produced maximum xylitol from corn cob acid hydrolyzed liquor with xylitol yield 87.716 g/L and volumetric productivity 0.018 g/L/h. CONCLUSION: Plant and agro-industrial biomass can be used as a carbohydrate source for the production of xylitol and ethanol after microbial fermentation. This study revealed that wheat straw acid and enzyme hydrolyzed residue proved to be best raw material for production of xylitol with S. cerevisiae. The xylitol produced can be utilized in pharmaceuticals after purification on industrial scale as pharmaceutical purposes.

Diagnosis of hepatitis C in pregnant mothers and its transfer pattern in neonates.

Hepatitis is the most common liver diseases in the Pakistan caused by blood-borne infection of HCV. Viral transmission is frequent through blood contact. Vertical transmission is transfer of disease from mother to infant. The women who are infected with hepatitis C virus RNA are at high risk of infecting their babies. Actual transmission occurs during labor and at time of delivery when blood of both mother and neonate is in contact with each other. Vertical transmission rate is lowered when mother is HCV RN A negative. The project was designed to determine the percentage of transmission and prevalence of Hepatitis C virus from mother to neonates. Assessment of the quantitative analysis of RNA levels in mother blood and viraemic status from the early postpartum period onwards of children born to HCV infected mothers. For the diagnosis of hepatitis C in mothers, blood samples of fifty HCV pregnant women between 23-41 years old were taken. The blood samples were centrifuged at 8,000 rpm and serum was separated and stored at 40C. The values of the Alanine Aminotransferase was determined at 2nd and 3rd trimester of pregnancy. After extraction HCVRN Awere transcribed and amplified by PCR. The samples were further authenticated through the Agarose Gel Electrophoresis system and bands were obtained. Nested reverse transcription PCR (RT-PCR) was conducted for the quantitative analysis of HCV-RNA. The results showed that in 66% cases, the mothers had high level of ALT at 2nd trimester of pregnancy. Their ALT level was decreased in the 3rd trimester of pregnancy. PCR results showed that 40% pregnant women had quantity of HCV-RNA in the range of 1000-10,000 IU/mL and in 18% women were above 100000 The results of spectrophotometer showed that 80% infants had the antibodies against HCV-RNA while only 20% of the neonates did not have antibody right after birth. The 29% babies got HCV-RNA in their serum and became positive for HCV-RNA.

Seed priming by sodium nitroprusside improves salt tolerance in wheat (Triticum aestivum L.) by enhancing physiological and biochemical parameters.

The germination, seedling vigor, crop establishment and yield of agronomically important crops is negatively affected by soil salinity. The current study aimed to investigate the ability of exogenous fertigation by sodium nitroprusside (SNP) to induce salt tolerance in four high yielding wheat cultivars (Sahar-06, Punjab-11, Millat-11 and Galaxy-13) that differ in their response to salt stress in terms of biomass production, oxidative defense mechanisms and grain yield. Three levels of SNP (0, 0.1 and 0.2 mM) were used for seed soaking. During soaking the seeds were kept in the dark. After soaking for 12 h the seeds were air-dried for 5 h before sowing. Salinity caused a significant reduction in biomass and grain yield, while it increased proline (Pro), ascorbic acid (AsA), hydrogen peroxide (H2O2) and malondialdehyde (MDA) contents. Cultivar Sahar-06 and Galaxy-13 were found more tolerant to salinity based on shoot length root fresh and dry wights, 100 grain weight, decreased MDA and H2O2 accumulation, phenolic and ascorbic acid (AsA) contents, accumulation of proline, activities of SOD, POD and CAT as compared to the other cultivars. Seed priming with SNP was effective in reducing the adverse effects of salt stress induced oxidative stress on plant biomass and grain yield in all the studied wheat cultivars, but maximum amelioration of salt stress tolerance by SNP treatment was found in cv. Sahar-06. The increased salt tolerance in wheat plants by SNP seed priming might be due to the role of NO in improving seed vigor and germination and early establishment of seedlings with better growth. 0.1 mM SNP was found the most effective in improving salt tolerance, as compared to other SNP concentations. Exogenous SNP fertigation increased the activities of antioxidant enzymes such as superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) and the contents of AsA, Pro and total phenolics content (TPC) in the salt stressed wheat plants. Our data indicate that SNP-priming induced salt tolerance by up-regulating the antioxidative defense mechanisms resulting in better biomass production and grain yield.