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Biosynthesized nanoparticles have played vital role recently, as suggested to be alternative to physical and chemical methods. In this study, biosynthesis of zinc oxide nanoparticles (ZnO NPs) were carried out using leaf extracts of Phoenix dactylifera L. and Zinc nitrate. The effect of ZnO nanoparticles on biomass and biochemical parameters was investigated. Biosynthesized ZnO nanostructure was characterized using X-ray diffraction (XRD), transmission electron microscopy (TEM), UV–visible spectrophotometer and Fourier transform infrared spectroscopy (FTIR). Which resulted in spherical shape with size ranging between 16 to 35 nm of Biosynthesized ZnO nanoparticles and UV absorption beak at 370.5 nm with clear peaks of functional groups. The impact of different concentrations (0.0 mg/L, 80 mg/L and 160 mg/L) of biosynthesized ZnO nanoparticles on biomass and bioactive compounds production of Juniperus procera in vitro was investigated. The results showed that, biosynthesized ZnO NPs (80 mg/L and 160 mg/L) concentrations were boosted the growth of J. Procera with significantly compared to non-treated plants in vitro. The highest concentration (160 mg/L) of ZnO NPs was enhanced the growth of plant at beginning period, one month later shoots became yellow and callus turned to be brownish. Moreover, the influence of ZnO NPs on phytochemical compounds in callus of Juniperus procera was examined using GC–MS analysis. The differences among treatments were recoded. Overall, zinc oxide nanoparticles substantially improved the growth of shoots and callus with increasing of biochemical parameters such as chlorophyll a, total phenolic and flavonoids contents, besides the total protein and, SOD, CAT and APX activity. ZnO NPs might be induced some phytochemical compounds as well as inhibit.

Objective To evaluate the efficacy of Transabdominal pre-peritoneal repair in the treatment of inguinal hernias Materials and Methods: This retrospective study was conducted in Surgical A Unit, Khyber Teaching Hospital from January 2015 to December 2019. A total 114 patients were included in the study. All of the patients underwent Laparoscopic Trans-abdominal pre-peritoneal(TAPP) repair. After six months follow up, their outcome variables were analysed. Results: Mean age of the study population was 41±7.8(SD) years(range 21-67 years). 104 patients had unilateral while 10 patients had bilateral hernias. 89(78.07%) patients had primary hernia while 25(21.92%) patients had recurrent hernia. Mean postoperative hospital stay was 1.21±0.3(SD) days. Umbilical port infection was noted in 2(1.75%) patients while no case of deep/ mesh infection was noted. Recurrence was noted in one case only. Chronic groin pain was noted in 5(4.38%) cases. Conclusion: TAPP is a safe and effective treatment modality for the treatment of primary and recurrent inguinal hernias.

Silver nanoparticles (AgNPs) are presently the most commonly generated engineered nanomaterials and are found in a wide range of agro-commercial products. The present study was designed to synthesize AgNPs biologically using Ochradenus arabicus leaves and investigate their effect on the morphophysiological properties of Maerua oblongifolia raised in vitro. Physicochemical methods (ultraviolet–visible spectroscopy, Fourier transform infrared spectroscopy, and transmission electron microscopy were performed for characterization and for obtaining microphotographs of the AgNPs. Shoots of M. oblongifolia (2–3 cm) grown in Murashige and Skoog medium supplemented with different concentrations of AgNPs (0, 10, 20, 30, 40, or 50 mg L−1) were used. Following 6 weeks of in vitro shoot regeneration, the shoot number, shoot length, leaf number, fresh weight, dry weight, chlorophyll content, total protein, proline level, and antioxidant enzyme activities of the plants were quantified. We found that 20 mg L−1 AgNPs increased the shoot number, shoot length, fresh weight, dry weight, and chlorophyll content of the plants. The maximum total protein was recorded in plants that were administered the lowest dose of AgNPs (10 mg L−1), while high concentrations of AgNPs (40 and 50 mg L−1) increased the levels of proline and the enzymes superoxide dismutase and catalase. Our results indicate that green-synthesized AgNPs may be of agricultural and medicinal interest owing to their effects on plants in vitro.

Novel, safe, and effective antilisterial agents are required in order to prevent Listeria monocytogenes infections and maintain food safety. This study synthesized silver nanoparticles (AgNPs) from the shoot extract of in vitro-grown Tamarix nilotica (TN) and characterized them using X-ray diffraction, Fourier transform infrared spectroscopy, UV-visible spectroscopy, dynamic light scattering, scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDXS), and transmission electron microscopy (TEM). We also assessed the antilisterial potential of the synthesized TN-AgNPs by determining the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against two strains of L. monocytogenes and L. innocua. TN-AgNPs (2×MICs) showed a significant decrease in growth in all Listeria test strains. Release of cellular content and cell morphology analysis of TN-AgNP-treated bacterial cells demonstrated the mechanism of bactericidal activity of AgNPs. In addition, TN-AgNPs induced a significant decrease in swimming motility (62–71%), biofilm formation (57–64%), and preformed biofilms (48–58%) in all Listeria test strains at sub-inhibitory concentrations. Microtitre plate assay results for biofilm inhibition were confirmed by SEM and CLSM visualization of TN-AgNP-treated and TN-AgNP-untreated Listeria test strains. TN-AgNPs also showed wound-healing activity in MCF-7 cells by inhibiting cell migration in a scratch plate assay. TN-AgNP-induced enhanced reactive oxygen species generation in treated cells could be a plausible reason for the biofilm inhibitory activity of AgNPs. TN-AgNPs having antilisterial, antibiofilm, and wound-healing properties can be effectively used to prevent L. monocytogenes infections in the food industry and healthcare.

The pancreatic β-cells sustain systemic glucose homeostasis by producing and secreting insulin according to the blood glucose levels. Defects in β-cell function are associated with hyperglycemia that can lead to diabetes. During the process of insulin secretion, β-cells experience an influx of Ca2+. Thus, imaging the glucose-stimulated Ca2+ influx using genetically encoded calcium indicators (GECIs) provides an avenue to studying β-cell function. Previously, studies showed that isolated zebrafish islets expressing GCaMP6s exhibit significant Ca2+ activity upon stimulation with defined glucose concentrations. However, it is paramount to study how β-cells respond to glucose not in isolation, but in their native environment, where they are systemically connected, vascularized, and densely innervated. To this end, the study leveraged the optical transparency of the zebrafish larvae at early stages of development to illuminate β-cell activity in vivo. Here, a detailed protocol for Ca2+ imaging and glucose stimulation to investigate β-cell function in vivo is presented. This technique allows to monitor the coordinated Ca2+ dynamics in β-cells with single-cell resolution. Additionally, this method can be applied to work with any injectable solution such as small molecules or peptides. Altogether, the protocol illustrates the potential of the zebrafish model to investigate islet coordination in vivo and to characterize how environmental and genetic components might affect β-cell function.

The major focus of this research was to investigate the effects of religiosity factor on internet addiction among young adults enrolled at college level. We adopted two instruments to gather the information including OK-religious attitude scale for Muslims developed and used by Ok, Uzeyir, and Internet Addiction Test prepared by Widyanto and McMurran. In total, 800 Muslim college students enrolled in four colleges at graduate level of southern Punjab Pakistan were chosen through multi-phase sampling. The subscales revealed more than .76 Cronbach alpha coefficients. The outcomes expressed positive role in case of DE conversion in world faith toward internet indications, whereas intrinsic religious orientations remained beneficial in decreasing internet usage. Students' anti-religion subscale demonstrates higher increase in becoming of internet addict; however, intrinsic religious orientations show significant decrease in using internet. Similarly, DE conversion in world faith view and Anti-Religion Scale indicate students' significant contributions in expecting them being internet addict. The study determines that the religiosity factor considerably illuminate the variances in developing internet addiction among the Muslim college adults with the direct effect of intrinsic religious orientation and indirect effect of anti-religion aspect.

The purpose of this study was to examine the influences of religiousness on the prevalence of Narcissistic Personality Disorder (NPD) among young adults. Prevalence of three forms of Allportian religious orientation, three forms of quest religious orientation and seven symptoms of NPD were examined through self-reported measures. 618 randomly selected Muslim students from the four public sector Pakistani universities participated in the study. Three research instruments comprising Religious Orientation Scale developed by Gorsuch and McPherson, Quest Scale developed by Batson and Schoenrade and Narcissistic Personality Inventory developed by Raskin and Terry were used to collect the data. All subscales demonstrated more than .70 Cronbach Alpha Coefficients. The findings demonstrate comparatively higher presence of intrinsic, extrinsic personal and extrinsic social religious orientations among the Pakistani Muslim young adults. The presence of NPD symptoms remains higher among the participants too. The study concludes that the religious orientations significantly explain the variances in the prevalence of NPD symptoms among the Muslim university students with the direct effects of intrinsic and extrinsic personal religious orientations and indirect effects of quest religious orientations.

The study was conducted to investigate the mixture of Alangium salvifolium extracts with distinguished organic solvents which were generally used to screen for antibacterial activity against Macrococcus brunensis, Bordetella hinzii, Morganella morganii, Salmonella bongori, Klebsiella oxytoca, Alcaligens faecalis, Achromobacter xylosoxidans, Comamonas testosteroni, Pseudomonas aeruginosa, and Pseudomonas plecoglossicida that obtained from seven fish pathogens. Three pathogens were elucidated in the study which is conducted by using SDS-PAGE to its localisation. The extracted solvents (Acetone) gave good and enhanced antimicrobial activity in their way. The present investigation is an essential step for developing plant-based drugs which are eco-friendly for the management of the pathogenic bacteria of a fish pathogen. Development of commercial formulation of botanicals used to enhance the probability of managing such pathogenic bacteria. Further investigations are necessary for developing commercial formulations based on fields, animal trails, and toxicological experiments. This research may be footprint towards managing the newer antimicrobial entities against the fish pathogen.

Zincoxide nanoparticles (ZnO NPs) are among the most produced and used nanomaterials worldwide, and in recent times these nanoparticles have also been incorporate in plant science and agricultural research. The present study was planned to synthesize ZnO NPs biologically using Ochradenus arabicus leaves and examine their effect on the morphology and physiology properties of Maerua oblongifolia cultured in vitro. ZnO NPs were characterized by UV–visible spectroscopy (UV–vis), X-ray diffractometer (XRD), Fourier transform infrared spectroscopy (FT-IR), and transmission electron microscopy, which demonstrated hexagonal shape nanoparticles of size ranging from 10 to 50 nm. Thus, the study uncovered an efficient, eco-friendly and simple technique for biosynthesis of multifunctional ZnO NPs using Ochradenus arabicus following growth of Maerua oblongifolia shoots in different concentrations of ZnO NPs (0, 1.25, 2.5, 5, 10, or 20 mg L−1) in Murashige and Skoog medium. Remarkable increases in plant biomass, photosynthetic pigments, and total protein were recorded up to a concentration of 5 mg L−1, at the same time, the results demonstrated a significant reduction in lipid peroxidation levels with respect to control. Interestingly, the levels of proline and the antioxidant enzyme catalase (CAT), superoxide dismutase (SOD), and glutathione reductase (GR) activities were increased significantly in response to all ZnO NP treatments. These findings indicate that bioengineered ZnO NPs play a major role in accumulation of biomass and stimulating the activities of antioxidant enzymes in plant tissues. Thus, green-synthesized ZnO NPs might be of agricultural and medicinal benefit owing to their impacts on plants in vitro.

In quest to develop effective anticancer drugs, a series of twelve heterocycles (1 − 12) embedded with oxopyrrolidine rings was synthesized. The compounds (1–12) were characterized by microanalysis, FT-IR, UV–Vis and 1H NMR spectroscopy, and mass spectrometry. Experimental DNA binding and computational docking studies were carried out to identify the biological targets of 1–12. The values of DNA binding constants of 1–12 ranged from 1.5 × 104 to 4.0 × 105 M− 1. The docking energies of the DNA-compound adducts varied from − 23.84 to − 30.96 kJ/mol, and the adducts were stabilized by H-bonding and hydrophobic attractions. The compounds preferred to enter minor grooves of DNA during adduct formation. Cytotoxicity studies of 1–12 were carried out towards both cancerous and normal cell lines. Compounds 4, 6, 7, 8, 9, 10, 11 and 12 at 1.0 nM concentration exhibited good anticancer activities with cell viabilities in the range of 19–110% against breast (MCF-7) and microglial (BV-2) cancer cell lines. The potency of 1–12 as future anticancer agents was ascertained by drug likeness using Lipinski's rule of five. It was observed that 1–12 obeyed the ‘Rule of Five’ with logP values

The purpose of this study was to investigate the influence of religious orientations on the prevalence of social media disorder among Muslim university students in Pakistan. A total of 686 students (59.0% females), enrolled in the three public-sector universities, participated in the study. The new indices of religious orientation revised and a short version of social media disorder scale were adapted to collect the required information. Both scales demonstrated high reliability coefficients. Results demonstrate significant differences in the religious orientations of disordered and non-disordered social media users. Findings support that the higher extrinsic and quest religious orientations enhance likelihood of being disordered social media users. The enhanced intrinsic religious orientation, however, decreases the chances of being disordered social media users. The implications of research findings in related academic and non-academic settings are discussed at the end of this paper.

Despite the numerous potential antimicrobial products and antibiotics in market, bacterial infections are still a main root of diseases and death throughout the globe. Due to the multidrug-resistant bacterial strains, there is an immediate need for the development of new types of biocidal materials that can counter these multidrug-resistant strains that harbor in different types of articles used in our everyday life. In this direction, attention has been turned toward nanotechnology that is gaining remarkable momentum in the present century due to its ability to modulate materials into their nanosizes having enhanced physical, chemical, optical, biological, and other properties. These nanoparticle (NP)-based materials have emerged as new types of antimicrobials in the field of antimicrobial chemotherapy. Especially metals such as Ag, Cu, Ti, and Zn and their oxides, in the form of NPs such as silver NPs, silver-based nanostructured materials, TiO2 NPs, TiO2 nanocomposites, zinc oxide NPs and nanorods, and copper NPs have the tremendous potential as antimicrobial agents. These NPs have emerged as potential antimicrobial agents and found diverse applications such as nanomaterial-based medical dressings, NP-coated medical devices, and nanoantimicrobial coatings in textile industry. This chapter therefore summarizes the various antimicrobial applications of such NPs in various fields, especially in medical, health care, and in textile industry as an nanoantimicrobial coatings.

The cadmium (Cd), a heavy metal, causes toxicity, which leads to hampering the growth and development of the plant. The molecular and biochemical approaches were used for the investigation of antioxidant system response and genotoxicity in date palm (Phoenix dactyliferaL.) cv. Sagai in pot experiment having Cd. The root length was more affected than the shoot length as more accumulation of Cd occurs in roots. Fresh weights of root and shoot were reduced significantly in treated plants as compared to the control. The proline content was increased at low concentration of Cd (300 µM-CdCl2) than the medium and high concentrations (600 and 900 µM-CdCl2), respectively. The thiobarbituric acid reactive substances (TBARS) content was increased at 600 and 900 µM-CdCl2compared to the plants treated at 300 µM-CdCl2and controls. Antioxidant enzymatic assay was performed under Cd stress and compared with control plants. The catalase (CAT) and superoxide dismutase (SOD) activities were found to be high in plants treated with CdCl2at 300 µM compared to at 600 and 900 µM-CdCl2, respectively. The genotoxicity of Cd was assessed using the inter-simple sequence repeat (ISSR) marker where all treated and control plants were clustered into three main groups based on genetic similarity.P. dactyliferaplants were found to be more divergent at high Cd stress as compared to control and plants treated at low concentration of Cd.

Background & Objective: Alpha (α) thalassemia is a hereditary disorder and is caused by deletions or mutations in globin genes. It is present in two clinically significant forms: hemoglobin Bart hydrops fetalis (Hb Bart) syndrome and hemoglobin H (HbH) disease. It is highly prevalent in South-East Asia or Mediterranean countries. The most common deletion reported in alpha thalassemia in Pakistani population was –α3.7 with a frequency of 8.3%, and the rare forms were –α4.2 (0.2%) and αααanti3.7 (0.9%). In our study, diagnosis of severe anemia cases without any α and β mutations or deletions were made by using extended alpha thalassemia deletions panel. The main objective of this study was to determine the prevalence and to study the spectra of alpha thalassemia gene deletions in beta thalassemia patients with the use of an extended panel including −−SEA, −−FIL, −−MED, −−20.5, −−THAI in addition to –α3.7, –α4.2 & -αααanti3.7. Methods: The samples were collected in ethylenediaminetetraacetic acid (EDTA) vacutainers. A total of 156 samples were analyzed for alpha thalassemia mutations. This cohort included 121 samples of beta thalassemia major, nine samples of beta thalassemia minor and 26 without any evidence of beta thalassemia mutations. DNA was extracted with Qiagen extraction kit. The primers for determination of different subsets of alpha thalassemia deletions were included. PCR amplification was performed and result interpreted on agarose gel. Results: Co-inheritance of alpha thalassemia (–α3.7, –α4.2) with homozygous beta thalassemia was detected in 30% cases of studied cohort (37 out of 121). The most common found was –α3.7 deletion (35/37) as single/double deletions or in combination with -αααanti3.7. In undiagnosed cases screened for beta thalassemia major, we found Mediterranean (–αMED) deletion at specifically 875 bp on agarose gel. This is distinctive finding in case of detecting –αMED instead of any other deletion from Pakistan. Conclusion: Alpha thalassemia deletions (–α3.7, –α4.2) are the common co-inherited deletions found in beta thalassemia major patients. On the basis of results, we propose an extended alpha thalassemia genetic mutation panel should be used for screening of children presenting with anemia with suspicion of haemoglobinopathy.

Depression, anxiety, and stress are among major psychological disorders being predominant in present day. This study proposed to analyze the role of Muslim religiosity in male students showing these mental indications. A sample including 723 Pakistani young adults enrolled at college level was randomly chosen. Muslim Religiosity Measurement Scale and Depression, Anxiety and Stress Scale were utilized to gather information. Discoveries uncover an inverse relationship between conduct and affiliation with the symptoms of mental disorders, anxiety and stress among the respondents. Results bolster the incorporation of religious dimensions in psychological wellness and mental well-being thought of young adults in Pakistan.

Wild plants can contain bioactive compounds with potential activity against disease-causing microorganisms. In the Kingdom of Saudi Arabia, there are many plant species that may have antibacterial, antifungal, or antiviral activities, among other properties. We extracted bioactive compounds with methanol as well as with water from leaves of Breonadia salicina, which is an endangered plant found in the wild in Saudi Arabia. These extracts were tested against the bacteria Bacillus subtilis, Pseudomonas aeruginosa, Shigella sonnei, Escherichia coli, and Staphylococcus aureus. Both extracts showed antibacterial activity against all of the microorganisms, and thus, B. salicina leaf extract has potential as an antimicrobial agent for the preservation of foods, instead of synthetic chemical compounds. We found that the methanolic leaf extract was more effective than the aqueous crude extract against B. subtilis, P. aeruginosa, and S. aureus.

Berardinelli-Seip congenital lipodystrophy (BSCL) syndrome is a rare genetic disorder caused by dysregulation of glycemic and lipid metabolism. We report five BSCL cases with typical clinical pictures and complications. These, to the best of our knowledge, represent the first case series from Pakistan. BSCL is characterized by marked atrophy of adipose tissue, acromegaly, acanthosis nigricans and tall stature. We could not perform genetics studies in any patient owing to non-availability of genetic laboratory in Pakistan. All the cases presented hypertriglyceridemia. One case developed hyperinsulinism controlled with metformin. There is no curative treatment and the current approach is low-fat diet and management of insulin resistance and diabetes. Recently published studies showed that leptin-replacement therapy is promising in the metabolic correction of complications of BSCL. This highlights the importance of further research in BSCL treatment.

Genetic diversity among and within 10 populations of Podophyllum hexandrum was analyzed for precise assessments of genetic structure for conservation planning of wild Indian May apple (Podophyllum hexandrum) in the Central region of Himalayas. Random Amplified Polymorphic DNA (RAPD) studies revealed variability among and within population. Twenty primer sets were evaluated, 18 primers resulted in low and poor polymorphism,where as two gave better results which were counted for final analysis. We found a higher podophyllotoxin levels among the populations. However, the existing variation in podophyllotoxin content among the populations may be due to geographical location, altitude and habitat. It was difficult to ascertain these variations as genetic variations. Whereas morphological variations may have impact on podophyllotoxin content detected in different populations of P. hexandrum. Hence we recommend that any further cultivation of this species require optimization of environmental factors in order to increase the rate of production of podophyllotoxin from any collected population.

The morphological features of Podophyllum hexandrum Royle, a ‘critically endangered’ medicinal herb and a source of podophyllotoxin, were studied in populations growing in different parts of the Kumaun region of the Indian Central Himalaya. Plant growth performance in terms of biomass accumulation and podopyllotoxin levels in the rhizomes collected from eleven natural populations (P1 to P11, altitude ranging from 2740 to 3350 m) were analyzed. Morphological features, e. g., plant height, stem diameter and leaf area were, in general, negatively correlated with an increase in the altitude. Maximum aboveground (8.46 g/individual) and belowground (48.18 g/individual) biomass values were recorded from a population (P9) at the lowest altitude (2740 m) and, in general, the species was found to perform better at the lower altitudes. The podophyllotoxin content of rhizomes ranged between 0.36 - 1.08 % (on dry wt. basis) in different populations, and a positive correlation was observed between podophyllotoxin content and an increase in the altitude.

PURPOSE To determine the role of a single cycle of topical azithromycin in preventing long-term trachoma complications in children from an area with endemic trachoma. DESIGN A longitudinal cohort study. METHODS Two hundred thirty-eight (n = 238) children with active trachoma were enrolled in the current study. They were aged 1 to 10 years, with trachomatous inflammation based on the simplified World Health Organization grading system. These children were identified out of a survey of 8600 children from 7 villages in Punjab, Pakistan, where trachoma was endemic. The studied patients with active trachoma were treated with a single regimen of azithromycin 1.5% eye drops, given twice daily for 3 days, and were followed up for 3 years. The long-term effects of this therapy were documented for the first time in an endemic area. RESULTS Two hundred fourteen children (90%) were present at 1-year follow-up with 10% dropout, and 178 (75%) were present at 3-year follow-up with 25% dropout. Of these, 23% and 11% had active trachoma at 1 and 3 years, respectively. Active trachoma was minimized from 100% at baseline to 11% at the end of 3 years. The reinfection rate was 4%. None of the patients developed trachoma-related ocular complications during the study period. CONCLUSIONS A single cycle of topical azithromycin can be used as an effective and safe option for treating active trachoma in children in endemic areas. This study also documented that a single cycle of topical azithromycin prevents trachoma-related ocular complications in an endemic area.

Cancer is the most dangerous disease to haunt the mankind in the world today. Generally, the overall cancer mortality rates are similar in both the sexes. The reasons for most of these deaths are inefficacy and failure of the current methods of treatments or the unavailability of treatment options. The researchers of the world are actively integrating nanotechnology of treating of various cancers. The development of smart nanocarriers is one of the most important innovations in this direction. The nanocarriers of the different materials are being developed to improve the efficacy of current treatments. The present article describes the role of nanotechnology in cancer treatment emphasizing cancer nanotherapy, nanocarriers for drug delivery, types and the mechanisms of the nanocarriers. Besides, the efforts are made to discuss the recent advances in the nanocarriers, current challenges and the future prospective.

Copyright: © 2014 Siddiqui BS, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. A Note on Anti-leishmanial, Spasmolytic and Spasmogenic, Antioxidant and Antimicrobial Activities of Fruits, Leaves and Stem of Morinda citrifolia Linn – an Important Medicinal and Food Supplement Plant

Background and Aim: The undergraduate and postgraduate medical students withstand persistent pressure during their studies. This cross-sectional study is designed with the aim to find out the incidence of stress among undergraduate medical students of different academic years, and explore the effect of gender and difference in ethnicity, cultural, and social background on perceived stress and their coping strategies. Methods: We used a questionnaire to assess the level of perceived stress among Saudi and Pakistani undergraduate students in medical schools. Results: Around 46% Umm al Qura undergraduate medical students were found to be stressed with average stress score ranging between 15 and 15.43. On the other hand, Karachi Medical and Dental College students showed increased incidence of stress (63%), but their average scores are more or less similar to that of Saudi students. Academic overload is the major stressor and cognitive, emotional, and behavioral issues are common in both medical schools. Prayers and socializing with friends are the main coping strategies observed at both places. Conclusion: Difference in ethnicity, cultural, and social background affects the incidence of stress but not the severity. Measures should be taken to reduce the academic stress. Developing a supportive environment, providing student counseling, and educating them about time management may play an important role in reducing the incidence of stress as well as its severity.

A single, open and non comparative study was conducted at Anwar Shah Trust for C.P. & Paralysis in collaboration with the Departments of Neurology and Neurosurgery, Children Hospital Lahore, Pakistan to evaluate the effects of ACULASER THERAPY in childern suffering from Cerebral Palsy (C.P.) and associated Neurological Disorders like epilepsy, cortical blindness, spasticity, hemiplegia, paraplegia, diplegia, quadriplegia, monoplegia, sensoryneural deafness and speech disorders. In all 500 children were treated and the data was gathered during a period of 4 years from December 2006 till December 2010. These children were further classified according to the type of C.P. (spastic, athetoid, mixed) they suffered from and associated Neurological Disorders. This article shows results in C.P. childern who were treated with ACULASER THERAPY for a minimum of 08 weeks and more or had minimum of 15 treatment sessions and more. This article also shows that those childern who were given a break in the treatment for 1 month to 1 year did not show any reversal of the signs and symptoms. Analysis of the data showed that out of 342 children with Spasticity and Stiffness 294 showed marked improvement showing 87% success rate, out of 252 children with Epileptic fits, there was a significant reduction in the intensity, frequency and duration of Epileptic fits in 182 children showing 72% success rate, out of 96 children with Cortical Blindness 60 children showed improvement accounting for 63% efficacy rate, out of 210 children with Hearing Difficulties, 126 showed marked improvement accounting for 60% improvement rate, out of 380 children with Speech Disorders 244 showed improvement reflecting 64 % improvement rate, out of 192 children with Hemiplegia 142 showed improvement in movement, tone and power accounting for 74% improvement rate, out of 152 children with Quadriplegia 104 showed improvement in gross and fine motor functions showing 69% success rate and out of 116 children with Paraplegia of lower limbs 88 showed improvement in weight bearing, standing and movement accounting for 76% improvement rate.

Objective: To investigate the haemostatic/oxidative changes and its association in patients of acute leukemia at the time of diagnosis. Study Design: Cross sectional and descriptive. Setting: Study was carried out in National Institute of Blood Disease and Bone Marrow Transplantation during 2011 to 2013. Material and Methods: Activity of Superoxide dismutase(SOD), Glutathione Reductase(GR), Total antioxidant status(TAS), Prothrombin Time, Activated Partial Thromboplastine Time, Fibrinogen, Factor VIII and Tissue Factor were evaluated in the hemolysate, sera, citrated plasma and leukemic cells of 150 patients with acute leukemia at the time of diagnosis in 40 healthy individuals. Results: The mean age of patients was 29.8 years.Activity of SOD, GR and TAS were significantly higher in patients of acute leukemia as compared to control ( Disclosures No relevant conflicts of interest to declare.

Introduction: Vitamin K dependent factors i.e II, VII, IX and X plays an important role in the clotting mechanisms of the human body. Deficiency or defect in any of these factors, Vitamin K or Gamma glutamyl Carboxylase (GC) and Vitamin K Epoxide Reductase (VKOR) involved in the metabolism of these factors will compromise the clotting mechanisms and can lead to the bleeding disorder. 28 rare single mutations in VKORC1 coding sequence have been reported so far. Objective: To investigate the biochemical, haemostatic and molecular basis of the Vitamin K dependent factor deficiency in Pakistani subjects. Setting: NIBD &BMT, University of Bonn and Children hospital. Method: For this purpose 10 subjects, 9 males and 1 female with age ranging from 7 months to 13years were included from selected region of Pakistan who had a history of bleeding disorder and low levels of all the Vitamin K dependent clotting factors. BT, PT, APTT, Factor II, VII, IX, X,Platelet count and fasted serum bile acids were analysed for exclusion of thromobocytopenia which cause of bleeding and bile acid for cholestatic liver disease respectively. DNA sequencing was done for the GCCX and VKOR encoding gene i.e. GCCX-1 and VKORC-1 respectively to understand the etiology at molecular level. Results: Platelet counts of all the subjects were normal only in one case it was mild low 100x103/ul. Mean± S.E.M value of platelet was 440x103 ± 67. Factor II, VII, IX and X mean levels were 12.7%, 6.9%, 11.2%, 7.1% respectively. Out of ten subjects mean bile acid levels were increased in 3 patients (20 µmole/l) while 3 had low levels (1.2 µmole/l). 3 novel mutations identified in GCCX-1 gene in those patients who had normal levels of bile acids. Conclusion: Out of ten only in three patients novel mutations were identified in GCCX-1 while other 6 patients had abnormal levels of bile acid may be a cause of vitamin K clotting factors deficiency.Better strategies for the diagnosis of vitamin K deficiency are needed. Disclosures Oldenburg: SOBI: Consultancy.

Introduction: Von Willebrand disease is a common genetically inherited bleeding disorder. Types of VWD classified on the basis of coagulation factor assays but it cannot characterize further subtypes or verdict the quality and quantity of VWF. Multimer analysis facilitates the classification and prerequisite parameter to accurate diagnosis of VWD. Objective: To characterize the type and subtypes of VWD type I and II on the basis of laboratory data and analysis of VWF multimer structure. Setting: NIBD Karachi, University of Bonn Germany, Children Hospital & Chughtai Lab Lahore, PAEC Islamabad and HMC Peshawar. Material And Method: VWF Ag, Ricof, FVIII, Glycoprotein Ib and multimer analysis of VWD plasma on 1.6% medium resolution SDS-agarose gel electrophoresis was carried out, and then VWF multimer were transferred on nitrocellulose membrane. Incubated nitrocellulose membrane with using polyclonal rabbit anti-human VWF antibody and Goat anti-rabbit IgG HRP conjugate then protein detected by immune reaction after evaluation of the membrane under luminescence. Results: We studied 57 index patients, 2(3.5%) type I, 4(7.01%) type II, 3(5.26%) unclassified and 48(15.78%) were type III vWD. In type I,II, unclassified and III mean vWF Ag was 52.28, 35.15, 54.5 , 5.7 IU/dl, vWF RCo: 19.5, 18, 38.8, 7.22 IU/dl, vWF Ag/ RCof: 0.655, 0.393, 0.74, 0.55, FVIII 69, 25.7, 61.7, 7.22 IU/dl and GpIb: 27.25, 19.8, 39.8, 4.1. Significant loss of large multimers in type II while multimers were subtle in 2 cases, normal pattern in type I and entirely absence of multimers pattern in case of type III observed. Conclusion: Types and subtypes of VWD classified on the basis of concentration and distribution of vWF multimer by functional and immunological assays but single test is not enough for appropriate diagnosis. We need molecular genetics for von Willebrand disease for further diagnosis. Disclosures Oldenburg: SOBI: Consultancy.

Introduction: Von Willebrand disease (VWD) type III is the second most common inherited bleeding disorder in Pakistan. It is caused by severe quantitative deficiency of plasma Von Willebrand factor (VWF). Absence of VWF increases the severity of disease and is caused by homozygous/ compound heterozygous mutations in Von Willebrand factor gene. Objective: The aim of study is to characterize molecular genetics of Von Willebrand type III in Pakistani population. Setting: NIBD & BMT Karachi, Chughtais lab, Children's Hospital Lahore, PAEC Islamabad and HMC Peshawar. Material And Method: In this cohort study of Von Willebrand disease (VWD) type III, Blood samples of 48 unrelated patients of type III VWD were collected in National Institute Of Blood Disease & Bone Marrow Transplant (NIBD) Karachi, Chughtais lab, Children's Hospital Lahore, PAEC Islamabad and HMC Peshawar. Genomic DNA was extracted from peripheral blood by QIAamp DNA Blood mini kit (Qiagen) and Exon specific PCR was done for VWF gene and Direct gene sequenced on automated ABI-3130 Genetic Analyzer (Applied Biosystems). Sequence Variations in VWF were checked on ISTH-SSC VWD homepage (http://www.vwf.group.shef.ac.uk/), Ensemble genome browser (http://www.ensembl.org/), VWFdb hemobase and biobase biological database (http://www.hgmd.cf.ac.uk/ac/index.php). We have adopted the nomenclature for numbering amino acids of Human genome variation society (HGVS). Statistics analysis was done SPSSv17. Results: Sequence analysis detected mutations in 46 (95.83%) out of 48 samples. VWD type III has led to identification of 28 cases (60.8%) homozygous and 18 (39.1%) were compound heterozygous. We have indentified total 31 Mutations distributed as 17 missense mutations (54%), 7 nonsense mutations (22%) 2 small deletions (6%) , 2 insertion mutations (6%) and 3 splice site mutations (9%).19 of these were newly mutations in this cohort study.. Further method multiplex ligation dependent probe amplification (MLPA) is needed for detection of large deletions in two patients. Nonsense c.3931C>T, p.Q1311*was founder mutation in Pakistani patients. Conclusion: In cohort study, missense mutations are detected as common in among patients and most of the mutations identified in this cohort were homozygous due to Consanguinity in the family of the patients. Disclosures Oldenburg: SOBI: Consultancy.

A number of antitumor and other bioactive compounds were previously isolated from Vernonia amygdalina Del. This study was designed to further isolate and characterize compounds of medicinal value that may also have antitumor activity from this edible and commonly available plant. Bioassay-guided fractionation of the leaf extract of V. amygdalina (MEVA) led to the isolation and characterization of a known compound, epivernodalol for the first time in this plant. Its structure was identified by spectroscopic methods including 1H-NMR, 13C-NMR, MS, UV and IR spectra. In vitro growth inhibitory and cytotoxic evaluation of MEVA, its fractions and epivernodalol against HT-144 (skin melanoma) cell line was carried out by the Sulforhodamine B (SRB) assay. The results showed that epivernodalol and the dichloromethane fraction of V. amygdalina were active against HT-144 (skin melanoma) cell line. Vernonia amygdalina Del. leaf extract yielded another cytotoxic which was active against skin cancer. © 2010 International Formulae Group. All rights reserved. Keywords: Epivernodalol, fractionation, characterization, sesquiterpenes, spectroscopy, melanoma

A single, open and non comparative study was conducted at Anwar Shah Trust for C.P. & Paralysis in collaboration with the Departments of Neurology and Neurosurgery, Children Hospital Lahore, Pakistan to evaluate the effects of ACULASER THERAPY in childern suffering from Cerebral Palsy (C.P.) and associated Neurological Disorders like epilepsy, cortical blindness, spasticity, hemiplegia, paraplegia, diplegia, quadriplegia, monoplegia, sensory-neural deafness and speech disorders. In all 250 childern were treated and the data was gathered during a period of 3 years from December 2003 till December 2006. These children were further classified according to the type of C.P. (spastic, athetoid, mixed) they suffered from and associated Neurological Disorders. This article shows results in C.P. childern who were treated with ACULASER THERAPY for minimum 6 weeks and more or had minimum of 15 treatment sessions and more. This article also shows that those childern who were given a break in the treatment for 1 month to 1 year did not show any reversal of the signs and symptoms. Analysis of the data showed that out of 171 children with Spasticity and Stiffness 147 showed marked improvement showing 87% success rate, out of 126 children with Epileptic fits, there was a significant reduction in the intensity, frequency and duration of Epileptic fits in 91 children showing 72% success rate, out of 48 children with Cortical Blindness 30 children showed improvement accounting for 63% efficacy rate, out of 105 children with Hearing Difficulties, 63 showed marked improvement accounting for 60% improvement rate, out of 190 children with Speech Disorders 122 showed improvement reflecting 64% improvement rate, out of 96 children with Hemiplegia 71 showed improvement in movement, tone and power accounting for 74% improvement rate, out of 76 children with Quadriplegia 52 showed improvement in gross and fine motor functions showing 69% success rate and out of 58 children with Paraplegia of lower limbs 44 showed improvement in weight bearing, standing and movement accounting for 76% improvement rate.