Conserved B and T cell epitopes prediction of ebola virus glycoprotein for vaccine development: An immuno-informatics approach.

Ebola virus (EBOV), a non-segmented single-stranded RNA virus, is often-most transmitted through body fluids like sweat, tears, saliva, and nasal secretions. Till date, there is no licensed vaccine of EBOV is available in the market; however, the world is increasingly vulnerable to this emerging threat. Hence, it is the need of time to develop a vaccine for EBOV to hinder its dissemination. The current study has been designed for identification and characterization of the potential B and T-cell epitopes using the Immuno-informatics tools, and it helped in finding the potent vaccine candidates against EBOV. Prediction, antigenicity and allergenicity testing of predicted B and T cells' epitopes was done as well to identify their potential as a vaccine candidate and to measure their safety level respectively. Among B-cell epitopes "WIPAGIGVTGVIIA" showed a high antigenicity score and it would play an important role in evoking the immune response. In T-cell epitopes, peptides "AIGLAWIPY" and "IRGFPRCRY" presented high antigenicity score, which binds to MHC class-I and MHC class-II alleles respectively. All predicted epitopes were analyzed and compared with already reported peptides carefully. Comparatively, Peptides predicted in the present study showed more immunogenicity score than already reported peptides, used as positive control, and are more immunogenic as compared to them. Peptides reported in the present study do not target only Zaire EBOV (ZEBOV), as in previous studies, but also other species, i.e. Tai Forest EBOV (TAFV), Sudan EBOV (SUDV), Bundibugyo EBOV (BDBV), and Reston EBOV (RESTV) and would bring the promising results as potent vaccine candidates.

Effects of biochar and zeolite soil amendments with foliar proline spray on nickel immobilization, nutritional quality and nickel concentrations in wheat.

Since Ni-rich soils are a threat to the environment, growing edible crops on Ni-rich soils can pose a serious risk to human, animal, plant and ecosystem health and, hence, is considered as a challenging task for the researchers. Contrarily, limiting the bioavailability of Ni in such soils upon the addition of suitable amendments cum foliar spray of proteinogenic amino acids having an objective to alleviate stress to crop plants can considerably reduce the environmental risk. In this pot trail, we substantiate the effects of biochar (BR) and zeolite (ZL) addition in the soil along with proline (PN) spray on the resistance, and stress responses of wheat against Ni as well as on Ni translocation and accumulation in wheat plants grown on a Ni-rich soil contaminated by electroplating effluent. The treatments, applied with and without PN spray, involved: no amendment; BR; ZL; and a concoction of both amendments (BR50%+ZL50%). We found that BR50%+ZL50% treatment significantly immobilized Ni in the soil, reduced its accumulation in the shoot, root, and grain, blocked membrane lipid peroxidation and showed an improvement in photosynthetic parameters, the status of antioxidant activities, grain biochemistry and grain yield, compared to the control. Interestingly, exogenous PN spray caused a significant additive effect on the aforementioned parameters in the wheat plants grown on BR50%+ZL50% treated soil. Our results involved a reduced Ni bioavailability in wheat rhizosphere due to BR50%+ZL50% in soil and, furthermore, the additive effect of PN spray to scavenging ROS, obstructing peroxidation of lipid membrane and, thus providing resilience to wheat plant against Ni stress. The suggested technique can make Ni-rich soils suitable for cultivation and production of high-quality food by minimizing Ni bioavailability and toxicity to plants.

Anorexia nervosa among teenage girls: Emerging or prevalent?

OBJECTIVES: To find out frequency of anorexia nervosa (AN) among teenage girls (TG) and to find out the knowledge and practice regarding anorexia nervosa among teenage girls. METHODS: A cross sectional study was conducted at higher secondary public school, Rawalpindi from June 2013 till December 2013. A sample of 100 female students of the age group 13-19 years were inducted by systematic sampling technique. Mixed pretested questionnaire was filled after informed verbal consent. Data was entered and analysed using SPSS version 20. RESULTS: Participants mean age was 15.81 ± 1.323 years. Mean weight, mean height and mean body mass index were found to be 50.34 ± 10.445 kg, 160.14 ± 7.846 cm and 19.675 ± 4.1477 kg/m(2) respectively. Anorexia nervosa was found in 42 (42%) teenage girls while 58 (58%) were not having anorexia nervosa. Sufficient knowledge and positive practice were found to be present in 57 (57%) and 49 (49%) respectively. Statistically no significant association was found between KP and AN (p=0.73). CONCLUSION: Anorexia nervosa is an emerging health concern in Pakistan. Anorexia prevalent behaviour was observed in almost half of the teenage girls.

Regular breakfast consumption associated with high intelligence quotient: Myth or Reality?

OBJECTIVE: To find the frequency of regular breakfast consumption among Pakistani teenagers while the other objective was to find out the association between breakfast consumers (BC) and intelligence quotient (IQ). METHODS: This comparative cross sectional study was conducted on 102 students of a Public School Rawalpindi from August 2013 to January 2014. Participants were categorised into two groups i.e. regular breakfast consumers (RBC) and irregular breakfast consumers (IBC) according to their breakfast habits. A standardized questionnaire of Wechsler Intelligence Scale for Childrenwas used for IQ assessment. Data was then entered and analysed in SPSS version 20. RESULT: Out of the 102 individuals with mean age 17.17 ± 0.631, 58(56.9%) were females and 44 (43.1%) were males. There were 63 (61.8%) RBC while 39 (38.2%) were IBC. Among RBC there were 7 (6.9%) in challenged, 5 (4.9%) were below average, 33 (32.4%) in average group, 14(13.7%) in above average and 4 (3.9%) in gifted group. While among IBC, there was 1 (1%) among the severely challenged, 3 (2.9%) in challenged, 8 (7.8%) in below average, 22 (21.6%) in average group, 4 (3.9%) in above average and 1 (1%) in gifted group. There was no significant association found between breakfast intake and IQ level among students (p=0.98). CONCLUSION: More than half of the students were having regular breakfast. There was no significant association found among breakfast consumers and IQ. However the IQ score was more among RBC as compared to IBC.

Knowledge, attitude and practice regarding folic acid deficiency; A hidden hunger.

OBJECTIVES: To find the Knowledge Attitude and Practice regarding Folic Acid Deficiency among Women of Child Bearing Age (WPCBA). To find out the Association of Education Level with Practice of Folic Acid in WPCBA. METHODS: A Descriptive cross sectional study (Knowledge Practice and Attitude) was conducted at Military Hospital and Combined Hospital Rawalpindi from September 2012 to February 2013. About 400 married females of age group 21-42 years were included by convenient sampling technique. After taking informed verbal consent, a closed ended interviewer administered questionnaire was filled. Data was entered and analyzed using SPSS version 20. RESULTS: Mean age of the respondents was 30.31 + 5.280 years. Illiterate and literate were 165 (41.25%) and 235 (58.75%) respectively. The knowledge regarding folic acid need was 172 (43%). Only 161 (40.25%) thought that folic acid deficiency in pregnant women results in abnormality in newborn. In pregnancy, 205 (51.25%) had received folic acid supplementation. Association between education level and practice of folic acid was significant (p= 0.009) at 95% confidence level. CONCLUSION: Knowledge regarding folic acid deficiency among WOCBA was low along with the poor attitude. Practice was also not satisfactory. Education status plays important role in preventing micronutrient deficiency.

Type A and Type B personality among Undergraduate Medical Students: Need for psychosocial rehabilitation.

OBJECTIVES: To find out the frequency of Type A and Type B personality among the students of Undergraduate Medical College. To find association between student year and personality type. METHODS: A descriptive cross sectional study was conducted at Undergraduate Medical College, Rawalpindi from Sept. 2012 till Feb. 2013. Among 500 sample size, 100 students from each MBBS year were inducted by probability systematic sampling technique. After taking consent from the institute and students, data was collected on BECK anxiety inventory (BAI) questionnaire. According to BAI scale, students were identified as Type A or B personality. Data was analyzed using SPSS version 20. To find association between student year and personality type, Chi-square test of significance with 95% confidence level was used. RESULTS: First, second, third, fourth and final year students had 5 (1%), 6 (1.2%), 11 (2.2%), (13 (2.6%) and 19 (3.8%) type A personality respectively. Among all the study participants (n=500), total number of type A was 54 (10.8%) and type B personality students were 446 (89.2%). Type A personality was 29 (11.6%) in female students (n=250) and 25 (10%) in male students (n=250). Association between student year and personality type was significant (p=0.010) at 95% confidence level. CONCLUSION: Type A personality students existed in every class and there was a gradual increase in the number of type A personality students from 1st year to final year in an undergraduate medical college of Rawalpindi. Significant association was observed in student year and type A personality.

Computational analysis and expression profiling of two-component system (TCS) gene family members in mango (Mangifera indica) indicated their roles in stress response.

The two-component system (TCS) gene family is among the most important signal transduction families in plants and is involved in the regulation of various abiotic stresses, cell growth and division. To understand the role of TCS genes in mango (Mangifera indica ), a comprehensive analysis of TCS gene family was carried out in mango leading to identification of 65 MiTCS genes. Phylogenetic analysis divided MiTCSs into three groups (histidine kinases, histidine-containing phosphotransfer proteins, and response regulators) and 11 subgroups. One tandem duplication and 23 pairs of segmental duplicates were found within the MiTCSs . Promoter analysis revealed that MiTCSs contain a large number of cis -elements associated with environmental stresses, hormone response, light signalling, and plant development. Gene ontology analysis showed their involvement in various biological processes and molecular functions, particularly signal transduction. Protein-protein interaction analysis showed that MiTCS proteins interacted with each other. The expression pattern in various tissues and under many stresses (drought, cold, and disease) showed that expression levels varied among various genes in different conditions. MiTCSs 3D structure predictions showed structural conservation among members of the same groups. This information can be further used to develop improved cultivars and will serve as a foundation for gaining more functional insights into the TCS gene family.

Comparative genomics of light harvesting chlorophyll (LHC) gene family and impact of chlorophyll-A contents under drought stress in Helianthus annuus.

Drought is one of the main environmental stressors that can alter the water status of plants; negatively affect growth, assimilation, and photosynthesis; and eventually reduce crop yield. We explored the dependence of drought tolerance traits on chlorophyll-A content. Local sunflower cultivars (FH-01, FH-628, FH-633, FH-572, and FH-653) were grown in pots and subjected to drought by withholding water for 10, 15, or 20 d. One month after germination, the leaves of the treated and non-treated plants were collected and subjected to biochemical analyses. Under different water stress levels, the levels of peroxidase (POD), superoxide dismutase (SOD), catalase (CAT), and proline increased, whereas those of chlorophyll-A decreased. Regression analysis clearly found that proline (-0.442), POD (-0.528), SOD (-0.532), and CAT (-0.814) have negative beta coefficient values. Phylogenetic analysis revealed that the LHC gene family is divided into six clades. Subcellular locations indicated that most LHC genes were located in the chloroplast; however, only few genes were present in the peroxisomes and endoplasmic reticulum. Our research found that Arabidopsis thaliana LHC genes were highly homologous to the LHC genes of Helianthus annuus. Furthermore, the LHC genes of both species are located in the chloroplasts; therefore, they play a role in photosynthesis and renewable energy production. This study opens a new horizon for discussing the role of chlorophyll-A in the drought-related traits of sunflowers.

Comparative genomic analysis of MYB transcription factors for cuticular wax biosynthesis and drought stress tolerance in Helianthus annuus L.

Sunflower is an important oil-seed crop in Pakistan, it is mainly cultivated in the spring season. It is severely affected by drought stress resulting in lower yield. Cuticular wax acts as the first defense line to protect plants from drought stress condition. It seals the aerial parts of plants and reduce the water loss from leaf surfaces. Various myeloblastosis (MYB) transcription factors (TFs) are involved in biosynthesis of epicuticular waxes under drought-stress. However, less information is available for MYB, TFs in drought stress and wax biosynthesis in sunflower. We used different computational tools to compare the Arabidopsis MYB, TFs involved in cuticular wax biosynthesis and drought stress tolerance with sunflower genome. We identified three putative MYB genes (MYB16, MYB94 and MYB96) in sunflower along with their seven homologs in Arabidopsis. Phylogenetic association of MYB TFs in Arabidopsis and sunflower indicated strong conservation of TFs in plant species. From gene structure analysis, it was observed that intron and exon organization was family-specific. MYB TFs were unevenly distributed on sunflower chromosomes. Evolutionary analysis indicated the segmental duplication of the MYB gene family in sunflower. Quantitative Real-Time PCR revealed the up-regulation of three MYB genes under drought stress. The gene expression of MYB16, MYB94 and MYB96 were found many folds higher in experimental plants than control. The present study provided the first insight into MYB TFs family's characterization in sunflower under drought stress conditions and wax biosynthesis TFs.

Anti-Parkinson's Activity of Tribulus terrestris via Modulation of AChE, α-Synuclein, TNF-α, and IL-1ß.

Tribulus terrestris (T.T.) is a rich source of flavonoids and saponins, which have been reported to have neuroprotective and antioxidant potential. The current study was planned to investigate the anti-Parkinson's activity of T. terrestris methanol extract (TTME). It was hypothesized that TTME possessed antioxidant potential and can ameliorate Parkinson's disease (PD) via modulation of α-synuclein, acetylcholinesterase (AChE), TNF-α, and IL-1ß. To test this hypothesis, in silico and in vivo studies were performed. The PD model in rats was prepared by giving haloperidol, 1 mg/kg, i.p. Rats were divided into six groups: control, disease control, standard, and treatment groups receiving TTME orally at 100, 300, and 1000 mg/kg dose levels for 21 days. Behavioral observations and biochemical analyses were done. The TTME modulatory effect on mRNA expression of α-synuclein, AChE, TNF-α, and interleukins in the brain homogenate was estimated by RT-PCR. Compounds detected in HPLC analysis disrupted the catalytic triad of AChE in in silico studies. Behavioral observations showed significant (p < 0.05) improvement in a reversal of catatonia, muscular strength, locomotor functions, stride length, and exploration in a dose-dependent manner (1000 >300 >100 mg/kg) of PD rats. Endogenous antioxidant enzyme levels CAT, SOD, GSH, and GPx were significantly restored at a high dose (p < 0.05) with a notable (p < 0.05) decrease in the MDA level in TTME-treated groups. TTME at a high dose significantly (p < 0.05) decreased the level of acetylcholinesterase. RT-PCR results are showing down-regulation in the mRNA expression levels of IL-1ß, α -synuclein, TNF-α, and AChE in TTME-treated groups compared to the disease control group, indicating neuroprotection. It is concluded that TTME has potential to ameliorate the symptoms of Parkinson's disease.

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.

Proportion of exposure of passive smoking in teenage group and symptoms precipitated after exposure to second hand smoke.

Exposure of passive smoking and second hand smoke (SHS) exposure in teenage group results in precipitation of several symptoms. Five hundred teenagers between ages of 13 - 19 studying in Army Public School and College Westridge, Rawalpindi, were selected for descriptive cross-sectional study from September 2012 till February 2013. Out of the total 500, 279 (55.8%) were exposed to passive smoking while 221 (44.2%) were not exposed to passive smoking. Out of the exposed 279 (55.8%) students, males were 132 (47.3%) and females were 147 (52.7%) with mean age of 15.06 ± 1.805 years. After exposure to SHS, multiple responses were observed. Statistical significant association was found between symptoms precipitation and exposure to SHS. Nose irritation was present in 84, while 195 did not complain of it (p = 0.002). Allergic symptoms were precipitated with SHS in 42, while not in 21 students (p < 0.001). Asthma attack was precipitated in 38, while 241 did not experience precipitation of asthma attack (p < 0.001). More cough was present in 168, while not in 111 students (p = 0.037). Nausea was present in 41, while not in 238 (p < 0.001). Nose irritation was seen in 84 and not in 195 students (p = 0.002). More than half of the teenagers had exposure to passive smoking. Statistical significant association was found between passive smoking exposure and precipitation of symptoms.

Antibiotic resistance of E. coli isolates from urine samples of Urinary Tract Infection (UTI) patients in Pakistan.

Drug resistance is becoming alarming with the passage of time worldwide in general and in third world countries in particular. Human urine specimens of patients of urinary tract infection at Sheikh Zayed hospital, Lahore, Pakistan were analyzed for drug resistance in Escherichia coli. A total of 69 Escherichia coli isolates from human urine specimens were obtained and screened for their antibiograms. A total of seven antibiotic resistance profiles were obtained with over 65% of the isolates showing multi-drug resistance. Very high resistance levels were detected against augmentin and gentamicin (87.5 &77.5 % respectively) while imipenem and tazocin recorded the least resistance levels (32.5% and 12.5% respectively) among the isolates.

Study of rust resistance genes in wheat germplasm with DNA markers.

Wheat is a vital dietary component for human health and widely consumed in the world. Wheat rusts are dangerous pathogens and contribute serious threat to its production. In present study, PCR-Based DNA Markers were employed to check the rust resistance genes among 20 wheat genotypes and 22 markers were amplified. NTSYS-pc 2.2 was used to calculate genetic diversity and Nei and Li's coefficients ranged from 0.55 to 0.95. Cluster analysis was obtained using UPGMA (Unweighted Pair Group Method of Arithmetic Average) algorithm. Maximum no. of genes (23) was amplified from TW-760010 genotype whereas minimum no of genes (14) were amplified from TW-76005 genotype. The data gained from present study open up new ways to produce new varieties by breeding rust resistant germplasm to avoid the economic and food loss and varieties with improved characteristics.

Insect resistance and risk assessment studies of advanced generations of basmati rice expressing two genes of Bacillus thuringiensis

Advanced generations of different transgenic lines of indica basmati rice (Basmati-370) expressing two unrelated Bt genes, cry1Ac and cry2A were evaluated for resistance to Yellow Stem Borer (YSB) and Rice Leaf Folder (RLF) under field conditions compared to control lines over three years (2003-2005). Homozygous lines were selected and analyzed for insect resistance, morphological, physiochemical properties and risk assessment studies. After artificial infestation of target insects, the transgenic plants showed significant resistance. Data were recorded in terms of dead hearts and white heads at vegetative and flowering stage respectively. Transgenic lines showed up to 100 and 96 percent resistance against yellow stem borer at vegetative and flowering stages, respectively. Natural damage of rice leaf folder was also observed during the year 2005. The transgenic plants were 98 percent more resistant as compared to untransformed control plants. Variations in some morphological characteristics, e.g., the average number of tillers, plant height and maturity were also observed. Transgenic lines produced 40 percent more grains than control plants. All these characteristics were stably inherited in advanced generations. The transgenic lines had no significant effect on non-target insects (insects belonging to orders other than Lepidoptera and Diptera) in field or under storage conditions. Chances of pollen-mediated gene flow were recorded at a rate of 0.14 percent.