Effects of azadirachtin on protein synthesis in specific tissues of the desert locust Schistocerca gregaria

Azadirachtin A, one of the most polar of the many terpenoids occurring in the the seeds of the neem tree (Azadirachta Indica) has long been known to be the most potent in terms of its toxic effects on insects. Although its exact primary mode of action remains unclear, much research has implied that it inhibits protein biosynthesis in various insect tissues. This then leads to many secondary effects, such as sterility, developmental failure and general loss of biological fitness. The aim of the work reported in this thesis was to choose two specific proteins, lysozyme and vitellogenin, which could be easily quantified, and synthesised mainly by the fat body. Previous work had suggested that the proteins synthesised by fat body were reduced by azadirachtin. It was hoped that the synthesis of both could be induced by appropriate chemical stimuli both in vivo and in vitro. The insect studied, the desert locust Schistocerca gregaria, is known to be sensitive to the effects of azadirachtin, and is sufficiently large to allow ease of tissue sampling. Preliminary studies with the uptake of [3H]-leucine into the proteins of fat body confirmed that aza reduced incorporation into the acid-preciptable proteins of the tissue and of the haemolymph by respectively 42% and 25% over a short time course. Injection of bacterial lipopolysaccharide (LPS) was found to double the amount of circulating lysozyme within 24 hours. Further results suggested that this originated in both the fat body and haemocytes. Subsequent studies concentrated on the fat body as it yielded more material. An incidental observation from this work was that lysozyme induction and release may be a general result of any mechanical or chemical insult to the locusts. Azadirachtin A (3mug/g body weight) largely eliminated the increase in lysozyme activity, reducing it to resting levels. Studies with isolated fat body confirmed that the effect of both LPS and azadirachtin could be shown to be direct effects on the fat body itself. In studying the production of vitellogenin, it was found necessary to estimate the protein by immunological means. Vitellin was purified from mature ovarian tissue, and was used to raise a polyclonal antiserum in rabbits. This antiserum was found to cross-react with vitellogenin, and was used as the basis for various immunological assays. An ELISA assay allowed quantitative assays of both vitellogenin in fat body and haemolymph, and of vitellin in ovaries. Over a period of a month after imaginal moult, the proportion of vitellogenin in the female haemolymph rose from undetectable below 8 days to a maximum of 70% of the total protein. Treatment of females at 15 and 19 days with azadirachtin reduced the amount of vitellogenin by 62% and that in ovaries by 73% by 48 hours after treatment. These effects in vivo could be largely reproduced in vitro with isolated fat-body, using methoprene, a Juvenile Hormone analogue, as the stimxilus. In conclusion, the results presented show that azadirachtin prevented at least part of the increased production, induced by appropriate factors, of two identified proteins. This is consistent with previous biological observations of the action of the terpenoid

Social Achievement goals and Academic Adjustment among College Students: Data from Pakistan

The study was conducted to investigate the social achievement goals and academic adjustment among college students in Pakistan. These variables were studied with gender and family system. The sample was consisting of 750 students from which 429 were boys and 321 were girls, age range was 15-20 years. Students form joint family system were 585 and 165 students were form nuclear family system. Student Adaption to College Questionnaire (SACQ) developed by Baker and Siryk (1984) and Social Achievement Goals Questionnaire (SAGQ) developed by Ryan and Shim (2006) were used to collect the data. After analyzing the questionnaire responses of the students, this research was highlighted the problems that the students may face while living in a joint family system. Results indicated that boys have more adaptation to college environment and have more social achievement goals as compared to girls. This research was also helping in knowing that the students from nuclear family system have more social achievement and adjustment to college environment as compared to students from joint family system

Methods of reconstitution to investigate membrane protein function

Membrane proteins are notoriously difficult to investigate in isolation. The focus of this chapter is the key step following extraction and purification of membrane proteins; namely reconstitution. The process of reconstitution re-inserts proteins into a lipid bilayer that partly resembles their native environment. This native environment is vital to the stability of membrane proteins, ensuring that they undergo vital conformational transitions and maintain optimal interaction with their substrates. Reconstitution may take many forms and these have been classified into two broad categories. Symmetric systems enable unfettered access to both sides of a bilayer. Compartment containing systems contain a lumen and are ideally suited to measurement of transport processes. The investigator is encouraged to ascertain what aspects of protein function will be undertaken and to apply the most advantageous reconstitution system or systems. It is important to note that the process of reconstitution is not subject to defined protocols and requires empirical optimisation to specific targets.The work in this manuscript was generously supported by a project grant (APP1085340) from the National Health and Medical Research Council (NHMRC) of Australia

Evaluation of Two Wheat Varieties for Phytotoxic Effect of Mercury on Seed Germination and Seedling Growth

Two varieties of wheat (Triticum aestivum var. Blue Silver and T. aestivum var. Punjab 85) were tested for the phytotoxic eff ect of mercury on seed germination and seedling growth. Seed were treated with 25, 50, 75 and 100 ppm of HgCl2 solution. Both varieties showed enhanced seed germination at all levels of mercury compared to untreated control. However, shoot, root, and seedling length was signifi cantly (P<0.05) reduced in both wheat varieties at all the levels of mercury treatments particularly at 100 ppm of mercury. Th e highest reduction was observed in root length rather than shoot and seedling length. Th e tolerance indices for both varieties showed that that var. Blue Silver was more tolerant to mercury toxicity than var. Punjab 85

EVALUATION OF ANTIOXIDANT AND ANTIMICROBIAL POTENTIAL OF TWO ENDANGERED PLANT SPECIES ATROPA BELLADONNA AND MATRICARIA CHAMOMILLA

Background: Plants are the natural source of antioxidants as well as antimicrobial compounds that has great potentials in pharmaceutical industry. In the present study, two medicinal plants Atropa belladonna and Matricaria chamomilla were collected from Northern areas of Pakistan. Materials and Methods: The extracts of the collected plants were obtained by microwave assisted extraction (MAE) with changing parameters, power level and time; methanol and ethanol were solvents used during extraction. The extracts of plants were tested against different bacterial strains. Results: It was observed that ethanolic extracts of Atropa belladonna has more significant antimicrobial activity against S.aureus than E.coli. In parallel, methanolic extract of Matricaria chamomilla showed greater significant antibacterial activity against S.aureus when compared with E.coli. In comparison, ethanolic extracts of Matricaria chamomilla has shown more significant results against S. aureus than E.coli (p≤0.05). Both plants had no antibacterial activity against S.typhi. The free radical scavenging activity observed by DPPH assay, indicate that both plants have antioxidant activity at all levels of concentrations in solvent tested during the present work. However, methanolic extracts had greater antioxidant activity when compared with ethanolic extracts. Conclusion: Present study is thus helpful in highlighting present potentials for antioxidant and antimicrobial properties in the selected plants

Improving community case management of diarrhoea and pneumonia in district Badin, Pakistan through a cluster randomised study--the NIGRAAN trial protocol

BACKGROUND: Diarrhoea and pneumonia contribute 30% of deaths in children under 5 in Pakistan. Pakistan\u27s Lady Health Workers Programme (LHW-P) covers about 60% of the population but has had little impact in reducing morbidity and mortality related to these major childhood killers. An external evaluation of the LHW-P suggests that lack of supportive supervision of LHWs by lady health supervisors (LHSs) is a key determinant of this problem. Project NIGRAAN aims to improve knowledge and skills of LHWs and community caregivers through supervisory strategies employed by LHSs. Ultimately, community casemanagement (CCM) of childhood pneumonia and diarrhoea will improve. METHODS/DESIGN: NIGRAAN is a cluster-randomised trial in District Badin, Pakistan. There are approximately 1100 LHWs supervised by 36 LHSs in Badin. For this study, each LHS serves as a cluster. All LHSs working permanently in Badin who regularly conduct and report field visits are eligible. Thirty-four LHSs have been allocated to either intervention or control arms in a ratio of 1:1 through computer-generated simple randomisation technique. Five LHWs from each LHSs are also randomly picked. All 34 LHSs and 170 LHWs will be actively monitored. The intervention consists of training to build LHS knowledge and skills, clinical mentorship and written feedback to LHWs. Pre- and post-intervention assessments of LHSs, LHWs and community caregivers will be conducted via focus group discussions, in-depth interviews, knowledge assessment questionnaires, skill assessment scorecards and household surveys. Primary outcome is improvement in CCM practices of childhood diarrhoea and pneumonia and will be assessed at the cluster level. DISCUSSION: NIGRAAN takes a novel approach to implementation research and explores whether training of LHSs in supervisory skills results in improving the CCM practices of childhood diarrhoea and pneumonia. No significant harm to participants is anticipated. The enablers and barriers towards improved CCM would provide recommendations to policymakers for scale up of this intervention nationally and regionally

Factors Associated with School Burnout in High School Students of South Punjab, Pakistan

The present study aimed to investigate the factors associated with school burn out among high school students in South Punjab. It was hypothesized that high achievement goals and achievement emotions negatively affect the school burnout among students. Data was collected from different high schools situated in Multan, Muzaffargarh, Bahawalpur and Rahim Yar Khan in South Punjab. The sample was consisted of 813 students from which 387 were females and 426 were males. The age range of participants was 14 to 16, with a mean age of 15.5 ± 0.85. The Achievement Goal Questionnaire, Achievement emotions Questionnaire (AEQ) and The Maslach Burnout Inventory–Student Survey were used as the data collection tools. Findings indicated that the investigated variables (i.e., students’ levels of school burnout, achievement goals, and achievement emotions) displayed the expected correlation patterns, with a negative association between high achievement goals, high achievement emotions and school burnout. Furthermore, these correlation patterns were confirmed for both female and male students. Findings revealed that females are higher in the level of performance approach, mastery approach, performance avoidance, class related emotions, learning related emotions and test emotions as compared to males and males are higher in emotional exhaustion, cynicism, and academic efficiency. Findings also indicate that on the family side students from nuclear family systems have higher in performance approach, performance avoidance and class related emotions whereas students from joint family system were higher in emotional exhaustion, cynicism, and academic efficiency. On the public and private school comparison results revealed that students from public schools have higher level of performance approach, mastery approach, class related emotions, learning related emotions and test emotions as compared to students of private schools

Evaluation of some biological activities of Abelia triflora R Br (Caprifoliaceae) constituents

Purpose: To investigate the antioxidant, anti-inflammatory, antidiabetic,  cardiovascular and cytotoxic activities of the leaf extract and major compounds isolated from Abelia triflora R. Br. (Caprifoliaceae)Methods: The chloroform soluble fraction of A. triflora leaves was subjected to several column chromatographic separations to isolate its constituents.  Anti-inflammatory and antioxidant activities were determined in terms of the ability to inhibit NF-kB, iNOS activity and lipoxygenase enzyme, and to decrease oxidative stress in HepG2 cells. Antidiabetic and cardiovascular activities were determined by screening for peroxisome proliferator-activated receptor alpha (PPARα) and PPARɣ agonistic activities. In vitro cytotoxic activity was determined against a set of four human cancer cell lines (SK-MEL, KB, BT-549, SK-OV-3) and two  non-cancerous kidney cell lines (LLC-PK1 and VERO). Cell viability was measured by neutral red assay.Results: Three triterpene acids were isolated from the chloroform fraction namely; ursolic acid (4), 2, 3-dihydroxy ursolic acid (5) and 2, 3, 21-trihydroxy ursolic acid (6). The results showed that ursolic acid exhibited potent inhibition of lipoxygenase enzyme and iNOS (inducible nitric oxide synthase) activity with IC50 (half-maximal inhibitory concentration) value of 13.0 μg/mL, compared to parthenolide positive standard (IC50, 0.3μg/mL); furthermore, it inhibited NF-kB (nuclear factor-kappa B) with IC50 of 25.0 μg/mL, compared to parthenolide (positive standard, (IC50, 0.5 μg/mL). Also, ursolic acid possessed the highest cytotoxic effect against the three cell lines, SK-MEL (IC50, 14.5 μg/mL), BT-549 (IC50, 16.0 μg/mL) and SK-OV-3 (IC50, 12.5 μg/mL). Only 2,3-dihydroxy ursolic acid activated PPARɣ (1.5-fold at 25 μM), compared to rosiglitazone (positive standard, 3.7 fold at 10 μM)Conclusion: Among the investigated compounds, ursolic acid exhibited the highest anti-inflammatory and cytotoxic activities, while 2,3-dihydroxy ursolic acid demonstrated antidiabetic activity via activation of PPARɣ.Keywords: Abelia triflora, Anti-inflammatory, Antidiabetic, Cardiovascular activity, Antioxidant, Cytotoxi

Factors Affecting Child Health: A Study of Rural Faisalabad

ABSTRACT In this paper an attempt is made to explore the effect of socio-cultural and democratic factors on the health of children. The study was conducted in rural tehsil Faisalabad, Pakistan. One hundred and twenty women of childbearing age 20 to 45 years with at least two living children under the age of 5 years were selected through systematic random sampling technique. A well structured questionnaire consisting of open-ended and close-ended questions was used for gathering information from the selected respondents. 84.2% of the respondents reported that their children are suffering from one or more of these diseases e.g. diarrhea, worm infestation, goiter, cholera and gastroenteritis and they also expressed about their poor health. Majority of the women were mal-nourished and anemic. The problems of antenatal and post-natal care were common in the selected area due to the unavailability and inaccessibility of the health care services along with traditional and dogmatic attitude towards the utilization of available limited health facilities. Delivery of baby under the supervision of untrained birth attendants at home was also responsible factor for high incidence of morbidity among children

Elicitation of the in vitro Cultures of Selected Varieties of Vigna radiata L. With Zinc Oxide and Copper Oxide Nanoparticles for Enhanced Phytochemicals Production

This study was conducted to develop a protocol for in vitro shoot multiplication and callus induction of various mung bean varieties to obtain enhanced phytochemical content with the help of elicitors. For shoot multiplication, two types of explants (shoot tips and nodal tips) of three varieties of mung bean (Mung NCM-13, MgAT-7, and MgAT-4) were used. Both types of explants from in vitro and in vivo sources were cultured on the MS medium supplemented with different concentrations (0.25–3.0 mg/L, increment of 0.5 mg/L) and combinations of BAP and IBA as independent treatments. For callus induction, leaf explants (in vitro source) were cultured on MS medium supplemented with 2,4-D (1–3 mg/L) alone or in combination with BAP or NAA (0.5 and 1.0 mg/L). For the enhanced production of phenolics and glycosides, calli were cultured on MS media supplemented with zinc oxide (0.5 mg/L) and copper oxide nanoparticles (0.5 mg/L) as nano-elicitors. Results showed that in vitro explants responded better in terms of shoot length, number of shoots, and number of leaves per explant when compared to in vivo explants. Moreover, shoot tips were better than nodal explants to in vitro culturing parameters. All three varieties showed the optimized results in the MS medium supplemented with 1 mg/L BAP, while roots were produced only in cultures fortified with 1 mg/L IBA. The leaf explants of in vitro and soil-grown plantlets showed a maximum callogenic response of 90 and 80%, respectively, on MS medium supplemented with 2,4-D (3 mg/ml). Maximum phenolic content (101.4 μg of gallic acid equivalent/g) and glycoside content (34 mg of amygdalin equivalent/g of plant material) was observed in the calli cultured on MS medium supplemented with 3 mg/L of 2,4-D. Furthermore, the addition of zinc oxide (0.5 mg/L) and copper oxide (0.5 mg/L) nanoparticles to the callus culture medium significantly enhanced the phenolic content of Mung NCM-13 (26%), MgAT-7 (25.6%), and MgAT-4 (22.7%). Glycosidic content was also found to be increased in Mung NCM-13 (50%), MgAT-7 (37.5%), and MgAT-4 (25%) varieties when compared to the control. It is suggested that elicitation of in vitro cultures of mung beans with nanoparticles could be an effective strategy for the enhanced production of secondary metabolites