Synergistic impact of -linolenic acid and -tocopherol on in vitro maturation and culture of buffalo oocytes

Abstract The objective of the current study was to investigate the synergistic impact of -Tocopherol and -Linolenic acid (100 µM) on IVM and IVC of Nili Ravi buffalo oocytes. Oocytes were obtained from the ovaries of slaughtered buffaloes within two hours after slaughter and brought to laboratory. Buffalo cumulus oocyte complexes were placed randomly in the five experimental groups included; GROUP 1: Maturation media (MM) + 100 µM ALA (control), GROUP 2: MM + 100 µM ALA + 50M -Tocopherol, GROUP 3: MM + 100 µM ALA + 100M -Tocopherol, GROUP 4: MM + 100 µM ALA + 200 M -Tocopherol and GROUP 5: MM + 100 µM ALA + 300 M -Tocopherol under an atmosphere of 5% CO2 in air at 38.5 °C for 22-24 h. Cumulus expansion and nuclear maturation status was determined (Experiment 1). In experiment 2, oocytes were matured as in experiment 1. The matured oocytes were then fertilized in Tyrodes Albumin Lactate Pyruvate (TALP) medium for about 20 h and cultured in synthetic oviductal fluid (SOF) medium to determine effect of -Linolenic acid (100 µM) and -Tocopherol in IVM medium on IVC of presumptive zygotes. To study the effect of -Linolenic acid (100 µM) in IVM media and increasing concentration of -tocopherol in the culture media on early embryo development (Experiment 3), the presumptive zygotes were randomly distributed into the five experimental groups with increasing concentration of -tocopherol in culture media. Higher percentage of MII stage oocytes in experiment 1(65.2±2.0), embryos at morula stage in experiment 2 (30.4±1.5) and experiment 3 (22.2±2.0) were obtained. However, overall results for cumulus cell expansion, maturation of oocyte to MII stage and subsequent embryo development among treatments remain statistically similar (P > 0.05). Supplementation of -tocopherol in maturation media having -Linolenic acid and/or in embryo culture media did not further enhance in vitro maturation of oocyte or embryo production.

Resumo O objetivo do presente estudo foi investigar o impacto sinérgico do -tocoferol e do ácido -linolênico (100 µM) na MIV e CIV de oócitos de búfala Nili Ravi. Os oócitos foram obtidos dos ovários de búfalos abatidos duas horas após o abate e levados ao laboratório. Complexos de oócitos cumulus de búfalo foram colocados aleatoriamente nos cinco grupos experimentais incluídos; GRUPO 1: Meio de maturação (MM) + 100 µM ALA (controle), GRUPO 2: MM + 100 µM ALA + 50 µM -tocoferol, GRUPO 3: MM + 100 µM ALA + 100 µM -tocoferol, GRUPO 4: MM + 100 µM ALA + 200 M -tocoferol e GRUPO 5: MM + 100 µM ALA + 300 M -tocoferol sob uma atmosfera de 5% de CO2 em ar a 38,5 °C por 22-24 h. A expansão cumulus e o estado de maturação nuclear foram determinados (Experimento 1). No experimento 2, os oócitos foram maturados como no experimento 1. Os oócitos maturados foram então fertilizados em meio de Tyrode's Albumina Lactato Piruvato (TALP) por cerca de 20 h e cultivados em meio de fluido oviductal sintético (SOF) para determinar o efeito do ácido -linolênico (100 µM) e -tocoferol em meio IVM em IVC de presumíveis zigotos. Para estudar o efeito do ácido -linolênico (100 µM) em meio IVM e aumentar a concentração de -tocoferol no meio de cultura no desenvolvimento inicial do embrião (Experimento 3), os presumíveis zigotos foram distribuídos aleatoriamente nos cinco grupos experimentais com concentração crescente de -tocoferol em meios de cultura. Maior porcentagem de oócitos em estágio MII no experimento 1 (65,2 ± 2,0), embriões em estágio de mórula no experimento 2 (30,4 ± 1,5) e experimento 3 (22,2 ± 2,0) foram obtidos. No entanto, os resultados gerais para a expansão das células do cumulus, maturação do oócito para o estágio MII e desenvolvimento embrionário subsequente entre os tratamentos permanecem estatisticamente semelhantes (P> 0,05). A suplementação de -tocoferol em meios de maturação com ácido -linolênico e / ou em meios de cultura de embriões não aumentou ainda mais a maturação in vitro de oócitos ou a produção de embriões.

Assessment of Dyslipidemia in Cases of PCOS and Its Association with Age and BMI

Objective:To determine the frequency of dyslipidemia in patients of PCOS in BVH Bahawalpur.Material and methods:This cross sectional study was conducted at Department of Pathology, Quaid-e-Azam Medical College, Bahawalpur. Total 286 patients with PCOS having age 18-35 years were selected for this study. Total 3 ml fasting blood sample was taken from every patient for total cholesterol, LDL, HDL & Triglycerides. Results:Mean age of the patients was 24.40 ± 5.367 and mean BMI was 21.01 ± 1.912. Out of 286 patients of polycystic ovarian syndrome (PCOS) dyslipidemia was found in 69 (24.13%) patients. Statistically significant association between dyslipidemia and age of the patients was noted with p value 0.000. Significant association between BMI and dyslipidemia was noted with p value 0.000.Conclusion:Findings of this study showed higherpercentage of dyslipidemia in cases of PCOS. Age of the patients and BMI was significantly associated with dyslipidemia.

Attachment Styles And Death Anxiety Among Pregnant Women In Pakistan

@#The present study was designed to investigate the attachment styles and death anxiety among pregnant women. In order to meet the study objectives sample of (n=62) was recruited from hospitals of Rawalpindi (i.e., Maryam Memorial and Cantonment Hospital) and Islamabad (i.e., Shifa Medicine Hospital and Poly Clinic). Age range of the sample was 18 to 45 years. Attachment styles were assessed by Experience in Close Relationship Revised-Questionnaire (ECR-R) and death anxiety was assessed with Death Anxiety Scale. The findings revealed that there was significant positive correlation between attachment styles and death anxiety (p<0.05) and it was found that pregnant women scored low on secure attachment style. Younger pregnant women were high on preoccupied, fearful and dismissing attachment style as compare to older age pregnant women. Women with first pregnancy scored high on anxious, dismissing, and fearful attachment style as compare to women with second and third pregnancy. Pregnant women had pregnancy loss feel more death anxiety as compare to pregnant women with no history of pregnancy loss. ASEAN Journal of Psychiatry, Vol. 22 (2): March 2021: 1-10.

Pilea umbrosa ameliorate CCl induced hepatic injuries by regulating endoplasmic reticulum stress, pro-inflammatory and fibrosis genes in rat

BACKGROUND@#Pilea umbrosa (Urticaceae) is used by local communities (district Abbotabad) for liver disorders, as anticancer, in rheumatism and in skin disorders.@*METHODS@#Methanol extract of P. umbrosa (PUM) was investigated for the presence of polyphenolic constituents by HPLC-DAD analysis. PUM (150 mg/kg and 300 mg/kg) was administered on alternate days for eight weeks in rats exposed with carbon tetrachloride (CCl). Serum analysis was performed for liver function tests while in liver tissues level of antioxidant enzymes and biochemical markers were also studied. In addition, semi quantitative estimation of antioxidant genes, endoplasmic reticulum (ER) induced stress markers, pro-inflammatory cytokines and fibrosis related genes were carried out on liver tissues by RT-PCR analysis. Liver tissues were also studied for histopathological injuries.@*RESULTS@#Level of antioxidant enzymes such as catalase (CAT), superoxide dismutase (SOD), peroxidase (POD) and glutathione (GSH) decreased (p < 0.05) whereas level of thiobarbituric acid reactive substance (TBARS), HO and nitrite increased in liver tissues of CCl treated rat. Likewise increase in the level of serum markers; alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP) and total bilirubin was observed. Moreover, CCl caused many fold increase in expression of ER stress markers; glucose regulated protein (GRP-78), x-box binding protein1-total (XBP-1 t), x-box binding protein1-unspliced (XBP-1 u) and x-box binding protein1-spliced (XBP-1 s). The level of inflammatory mediators such as tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and monocyte chemoattractant protein-1 (MCP-1) was aggregated whereas suppressed the level of antioxidant enzymes; γ-glutamylcysteine ligase (GCLC), protein disulfide isomerase (PDI) and nuclear erythroid 2 p45-related factor 2 (Nrf-2). Additionally, level of fibrosis markers; transforming growth factor-β (TGF-β), Smad-3 and collagen type 1 (Col1-α) increased with CCl induced liver toxicity. Histopathological scrutiny depicted damaged liver cells, neutrophils infiltration and dilated sinusoids in CCl intoxicated rats. PUM was enriched with rutin, catechin, caffeic acid and apigenin as evidenced by HPLC analysis. Simultaneous administration of PUM and CCl in rats retrieved the normal expression of these markers and prevented hepatic injuries.@*CONCLUSION@#Collectively these results suggest that PUM constituted of strong antioxidant chemicals and could be a potential therapeutic agent for stress related liver disorders.

Effect of kisspeptin on serum prolactin and gonadotropins levels in Balb-c mice with induced chronic stress

Objective: To determine and compare effects of kisspeptin on serum prolactin, luteinizing hormone and follicle stimulating hormone levels in Balb-c mice with and without exposure to chronic restraint stress

Study Design: An animal experimental study

Place And Duration Of Study: Shifa College of Medicine / Shifa International Hospital Islamabad, in collaboration with National Institute of Health, Islamabad and Centre for Research in Experimental and Applied Medicine Laboratory, Army Medical College, Rawalpindi, from April 2014 to June 2015

Methodology: Mice divided into three groups, each containing 30 mice. Control group [group A] received intraperitoneal injection of saline, group B was administered with intraperitoneal injection of saline and restrained stress, and group C was administered with both stress and kisspeptin 100 ng daily for four weeks. Restraint stress was applied to groups B and C for three hours per day by immobilising individual mice in wire-mesh restrainers. At the end of four weeks blood sampling was done. Serum luteinizing hormones [LH], serum follicular stimulating hormone [FSH] and serum prolactin [PRL] were analysed by ELISA

Results: Serum prolactin level increased in group B [stressed] and group C [stressed + kisspeptin treated] as compared to control group; and decreased in group C as compared to group B. Serum LH and FSH in group B was decreased as compared to control, and it was increased in group C as compared to control and group B

Conclusion: Administration of kisspeptin increases level of gonadotropins and reduces stress-induced hyperprolactinemia, which may improve fertility despite stress in animal

Poverty-associated emerging infection of Cystic Echinococcosis in population of Northern Pakistan: A hospital based study

@#Cystic Echinococcosis (CE) is one of the most important zoonotic parasitic diseases in human, livestock, and wildlife globally. The prevalence of CE depends upon human behavioral risk factors, the diversity and ecology of animal host interactions and the genetic diversity within Echinococcus species which differ in their zoonotic potential and pathogenicity. It is a neglected, economic and socio-cultural problem in Pakistan. The available data about the incidence of CE is very limited and no extensive study has been reported in Pakistan. The current study was aimed to analyze the hospital reported cases of CE and the associated risk factors related to the incidence of CE. The hospital-based data of CE for the time period of January 2012-December 2017 was collected from Islamabad, Rawalpindi and Peshawar. The data covered demographic characteristics including age, gender, and cyst localization of infected individuals and socioeconomic determinants. The data was analyzed based upon different risk factors along with the different socioeconomic parameters that has an important impact on the distribution of disease. A total of 228 cases were presented in the selected hospitals of different cities during the study period. Out of total 228 patients, 59.21% were males and 40.78% were females (P<0.001). Most infections have been recorded in young adults (>20-30) showing 22.8% of total infected individuals followed by children (0-10) showing 10.5% (n=24), respectively (P<0.001). Liver was the most vulnerable organ (58.77%, n=134) followed by lungs (14.47%, n=33) (P<0.001). The infection was higher among rural communities (84.2%) than urban (12.8%) (P<0.001). Socioeconomic and demographic factors had an important impact on the intensity of disease (P<0.001). The occurrence of cases in children and young adults was an important finding as it indicated an active transmission of the parasite in Pakistan along with the poverty index. Emergence of echinococcosis in Pakistan showed that emerging health issues in Pakistan could bring the disease to limelight for future research. This finding, together with the fact that 1 hospital reported 214 cases over 6 years 325 underlines the need for a program for prevention/control of this disease in Pakistan. The timely measure needs to be taken to hamper the disease development and establishment. In order to control the disease, complete surveillance should be done which in turn weighs down the disease progress.

Identification and Characterization of Two New S-Adenosylmethionine-Dependent Methyltransferase Encoding Genes Suggested Their Involvement in Stipe Elongation of Flammulina velutipes

Two new SAM-dependent methyltransferase encoding genes (fvsmt1 and fvsmt2) were identified from the genome of Flammulina velutipes. In order to make a comprehensive characterization of both genes, we performed in silico analysis of both genes and used qRT-PCR to reveal their expression patterns during the development of F. velutipes. There are 4 and 6 exons with total length of 693 and 978 bp in fvsmt2 and fvsmt1, respectively. The deduced proteins, i.e., FVSMT1 and FVSMT2 contained 325 and 230 amino acids with molecular weight 36297 and 24894 Da, respectively. Both proteins contained a SAM-dependent catalytic domain with signature motifs (I, p-I, II, and III) defining the SAM fold. SAM-dependent catalytic domain is located either in the middle or at the N-terminal of FVSMT2 and FVSMT1, respectively. Alignment and phylogenic analysis showed that FVSMT1 is a homolog to a protein–arginine omega-N-methyltransferase, while FVSMT2 is of cinnamoyl CoA O-methyltransferase type and predicted subcellular locations of these proteins are mitochondria and cytoplasm, respectively. qRT-PCR showed that fvsmt1 and fvsmt2 expression was regulated in different developmental stages. The maximum expression levels of fvsmt1 and fvsmt2 were observed in stipe elongation, while no difference was found in mycelium and pileus. These results positively demonstrate that both the methyltransferase encoding genes are involved in the stipe elongation of F. velutipes.

Doppler ultrasound evaluation of cerebral blood flow in anaemia of chronic renal failure

Objective: To determine mean cerebral blood flow in anemia secondary to chronic renal failure by doppler ultrasound and to compare the results with data gathered from healthy control subjects. Study Design: Case control study. Place and Duration of Study: Combined Military Hospital, Lahore, from Jan 2011 to Jun 2011

Patients and Methods: A total of 60 subjects were included who were divided into two groups. Group I had 30 patients with anemia due to chronic renal failure while group II consisted of 30 healthy control subjects. Doppler for both groups was performed with a 7.5-MHz transducer of color doppler machine ALOKA SSD-5500 in a dimly lit room with a comfortable temperature [22°C-24°C] after an adaptation period of at least 15 minutes rest, in supine position. The internal carotid and vertebral arteries on both sides were examined in both groups and analyzed

Results: Patients with anemia due to chronic renal failure were found to have increased cerebral blood flow as compared to normal healthy individuals

Conclusion: Cerebral blood flow increases in patients with anemia due to chronic renal failure

Audit of pediatric surgical emergency: need for self evaluation

Background: The clinical or surgical audit is a systematic process in which the standard of clinical care being provided to the patients is sought through review of clinical records against specific criteria. The clinical or surgical audit is a mandatory element of the professional competence in many countries

Objective: To evaluate the yearly performance and to establish the pattern and frequency of various Pediatric Surgical emergencies encountered

Methodology: This was an observational, descriptive study covering the year 2016 with retrospective data collection was carried out in the Department of Pediatric Surgery Emergency Mayo Hospital/King Edward Medical University, Lahore

Results: A total of 3880 patients including 2350[60.6%] males and 1530[39.4%] females were admitted during the study period. The highest number of admissions were Burn Patients with the percentage of 37.8%, followed by trauma cases 23.9%, and acute appendicitis 18.6%. There had been 86 deaths which gave rise to 2.2% Mortality

Conclusion: Burn injuries and trauma are common injuries in pediatric surgery ward. It is suggested that audit must be conducted routinely and should be officially declared as a compulsory action to provide vision and feedback to the working of surgeons

Evaluation and assessment of prescribing patterns in elderly patients using two explicit criteria based screening tools: [the priscus list and stopp/start criteria]

Background and Objectives: Rational prescribing can prevent medication errors and the associated harm, especially in old age patients, as they are being frequently prescribed with drugs for various ailments .Moreover, polypharmacy is a common practice in them. Therefore, a significant threat of potential drug interactions and adverse effects exist. Current study focuses on assessment of Potentially Inappropriate Medication [PIM] in medication prescribed to old age patients

Methods: It was a forty days, descriptive and observational study conducted from August 15th 2017 to September 25th 2017 in which prescriptions given to elderly patients were reimbursed for collecting various sets of information. In order to assess PIMs [in Pakistani Set-up], STOPP/START addition 2008 [including examples of misprescribing, overprescribing and under prescribing] and the PRISCUS list [misprescribing and overprescribing] was used. Statistical analysis of results was performed using SPSS version 20

Results: One hundred forty six cases of PIMs [including incorrect prescribing, overprescribing and under prescribing] were detected. It included incorrect prescriptions 104, under prescription 28 and over prescriptions 14. NSAIDS accounted for most incorrect prescriptions followed by benzodiazepines. Mostly NSAIDS were used for myalgia, backache and rheumatoid disorders

Conclusion: Current findings highlighted Potentially Inappropriate Prescribing [PIP], particularly of NSAIDs and under prescribing of statins in cardiovascular diseases. Study findings suggest introducing pertinent interventions at the stages involved in prescribing, prescription review and its follow up to reduce the PIP and PIMs

computational structural analysis of functional attributes of hypodermin A and B proteins: a way forward for vaccine development

Hypodermosis is a parasitic disease of cattle. The pathogenicity of the disease is attributed to Hypodermin proteins [Hypodermin A, Hypodermin B and Hypodermin C]. Studies suggest that Hypodermin proteins may be defined as Serine proteases and collagenases. The structure of both proteases Hypodermin A and Hypodermin B were modeled using the Swiss-model server followed by its validation using Procheck, Errat and Verify-3D. Afterwards, both Hypodermin A and Hypodermin B were docked against collagen in order to study its interaction with respective Hypodermin proteins. The structure of both Hypodermin A and Hypodermin B showed more bent towards hydrophobic nature, as more beta sheets were present in them. Both structures were also superimposed to check out similarities and differences present between them. Serine, Aspartic acid, Histidine, Glutamic acid and Lysine are found as interacting residues that are involved in hydrogen bonding with collagen. The interactions are found in the active domain region of Hypodermin proteins. The interacting residues were present in the active region of the hypodermin proteins thus validating the docking studies. This study may help in the drug development against hypodermosis with least side effects

Reduction of body and testicular weight of Albino rats in Arsenic toxicity

Abstract: The metals toxic for living being are led, mercury, arsenic, cadmium and cobalt. Arsenic is increasing day by day in underground water and is more toxic for the proliferating tissue in the body as intestine, ovaries, testes etc

Study design: Experimental study

Place and duration: National Institute for health, Islamabad from 2008 to 2014

Methodology: 20 Male albino rats were bought from National Health Institute, Islamabad weighing from 250-300 grams. They were divided in to two groups A and B each having 10 animals. Group A was taken as control and was given 10 ml of distilled water orally in the morning and evening with normal diet for 28 days. Group B was given 10 ml of water with sodium arsenite 5mg/kg of body weight for 28 days with normal diet

Results: The present study showed that there was reduction of body and testicular weight of albino rats given arsenic when compared with control

Conclusion: Arsenite and arsenate compounds are highly toxic to human beings as well as animals. Arsenic induced toxicity might be responsible for regression of testes and reduces the body and testicular weight of albino rats

A Retrospective Analysis on the Cystic Echinococcosis Cases Occured in Northeastern Punjab Province, Pakistan

A retrospective study was performed to report the case occurrence of cystic echinococcosis (CE) in three hospitals of north-eastern region in Punjab Province, Pakistan. We reviewed retrospectively the clinical records of patients in 4 hospitals which were diagnosed with CE during 2012–2017. A total of 198 cases, 82 (41.4%) male and 116 (58.6%) females were detected as CE. The most Highest incidence was revealed in.... 21–30 years-old group (24.2%) followed by 41–50 (22.7%), 31–40 (16.2%), 11–20 (13.6%), 51–60 (8.1%), below 10 (5.5%), over 71 (5.1%) and 61–70 year-old group (4.5%). CE was detected in various organs of infected individuals. However, most of CE cases were detected in the liver (47.4%) and lung (18.6%). The present study indicated that CE is more or less prevalent in surveyed areas and one of the most important public health problems in Punjab Province, Pakistan.

A study of knowledge and attitude of health care providers working at tertiary care hospitals of Lahore, Pakistan (having HIV/AIDS treatment facility) towards HIV/AIDS

A very few health care providers look after HIV/AIDS people because of their poor knowledge and either their lack of positive attitude or usually both. The rationale of this study was to evaluate the knowledge and attitudes of health care providers (tertiary care hospitals in Lahore, having HIV/AIDS Treatment Facility) towards patients with HIV/AIDS. In this cross sectional study, a random sample of 180 healthcare providers was assessed for their knowledge and attitudes. In the questionnaire of knowledge, items were measured in the form of dichotomous scale. Whereas in the five domains of attitudes, items were measured as a five-point likert scale ranging from strongly disagree to strongly agree. Results were discussed in the outlook of two variables which showed that the presence of negative attitude and lack of awareness increase the level of stigma and discrimination. HIV in our country needs positive attitude of rendering care and effective control measures by creating awareness in the community. The study reinforced the need for an ongoing education focused on experiential learning, and professional socialization.

Non-invasive investigations in heroin users and non-users - a case control study

Objective: To compare heroin users with their age and gender matched relatives as regards non-invasive investigations like chest x-ray, electrocardiography [ECG] and 2D-echocardiography

Study Design: Case control study

Place and Duration of Study: Armed Forces Institute of Cardiology-National Institute of Heart Diseases, Rawalpindi, from Feb 2010 to Jul 2010

Material and Methods: Sixty one heroin users [cases] and their 100 non-substance abuser age and gender matched relatives [controls] were included through non-probability consecutive sampling. Chest x-ray [CXR], 12-lead electrocardiography [ECG] and two-dimensional echocardiography [2D-ECHO] were done of each one and the result was interpreted by respected consultants. The data were analyzed on SPSS version 15. Descriptive statistics of mean and standard deviation [SD] were described for quantitative variables while frequency and percentages for qualitative variables. Independent samples t-test compared quantitative variables while for qualitative variables chi-square test was used. Ninety five percent confidence intervals were calculated for all variables

Results: Statistically significant differences were noted between cases and controls as regards chest X-ray. A total of 10% cases had findings consistent with tuberculosis [TB] as compared to 3% controls [p=0.045]. There was however no cardiac abnormality in any study subject. Significant 12-lead ECG findings other than normal were noticed in 12 [19.7%] heroin users as compared to 10 [10%] controls [p=0.083]. In 12 lead ECG, the predominant finding was t wave inversion in leads AVF and III. According to 2D-echocardiography [2D-echo] reports, 15 [24.6%] heroin users [cases] and 17 [17%] controls had abnormal findings [p=0.242]. We report no vegetations on any cardiac valve

Conclusion: On radiological and cardiac parameters, heroin users are a different population as compared to the non-heroin users. The knowledge about variations on non-invasive investigations not only helps the healthcare workers to optimize treatment regarding rehabilitation but also facilitate diagnosis and treatment

Frequency of headache with 25G or 27G quincke needles after spinal anesthesia in patients undergoing elective cesarean section

Objectives: Post-dural puncture headache [PDPH] is an iatrogenic complication of spinal anesthesia which results from puncture of the dura mater. The signs and symptoms of PDPH are thought to be caused due to loss of cerebrospinal fluid, traction on the cranial contents and reflex cerebral vasodilation. The patient's age, sex and the size of the dural perforation are the two most important factors affecting the frequency and severity of PDPH. We aimed to compare the frequency of post-dural puncture headache [PDPH] with 25G quincke needle and 27G quincke needle in spinal anesthesia in patients undergoing elective cesarean section

Methodology: This randomized controlled trial was carried out in our anesthesiology department over a period of six months from 1st December 2014 to 30th May 2015. A total of 124 parturients, ASA physical status I to II undergoing cesarean section were included in this study. Patients with infection at the site of injection, severe hypovolemia, coagulopathy, raised intracranial pressure, severe aortic and mitral stenosis, severe preeclampsia, placenta previa grade II-IV, placenta accreta and twin pregnancy were excluded from the study. Enrolled parturients were divided into two groups A and B, with 62 patients in each group. In Group-A and Group-B, Quincke spinal needles 25G and 27 G were used respectively to administer spinal anesthesia in the sitting position at the L3-4 or L4-5 intervertebral spaces. PDPH was assessed after 6, 12, 24 and 48 hours of surgery. Qualitative variable are presented as frequency and percentages while quantitative variable like age was presented as mean +/- SD. Sample t-test was applied on quantitative variable age. Chi-square test was applied for comparison of PDPH. A P- value < 0.05 was considered statistically significant

Results: The ages of patients were between 18 to 40 years. Mean age of the patients in Group-A was 27.77 +/- 4.82 y and 27.74 +/- 4.30 y in Group-B. Nine patients [14.5%] of Group-A experienced PDPH as compared to 2 patients [3.2%] of Group-B. There was statistically significant difference [p = 0. 027] between the groups

Conclusion: We conclude that 27G quincke spinal needle has definite advantage over 25G quincke spinal needle in terms of frequency of PDPH in spinal anesthesia for cesarean sections

Surgical outcome of spelenectomy in Thalassemia major in children

Objective: To determine the surgical outcome of splenectomy in children with thalassemia major

Methods: It is an observational and descriptive study conducted in Department of Paediatric Surgery in collaboration with hematology, radiology, anesthesia and paediatric intensive care department at The Children's Hospital and the Institute of Child Health, Multan during the period of September 2007 to September 2013. A total of 50 patients suffering from thalassemia major already diagnosed and under management reffered from haematology department for splenectomy were included in this study. After admission, patients were assessed on the basis of history, clinical examination, and necessary investigations before surgery and later on follow-up. Investigations carried were CBC, PT, APTT, Viral markers, ECG, X-ray Chest, abdominal ultrasonography and ECHO if necessary. Splenectomy was performed after prophylactic vaccination against post splenectomy infections. Follow up was performed for at least two years. Blood transfusion requirements and number of hospital visits per annum before and after splenectomy were calculated and results analyzed statistically using SPSS-20

Results: Fifty patients were included in this study. Out of these fifty, 43 [86%] male and 7[14%] were female with a mean age of 9 years. Average blood transfusion requirement was 250 ml/kg/year, interval of blood transfusion was two weeks and twenty five visits per year before splenectomy. After splenectomy, requirement of blood transfusion reduced to 125ml/kg/year, interval between transfusion increased to one month and hospital visits reduced up to twelve per year

Conclusion: Blood transfusion requirement and number of hospital visits per year are decreased and interval between transfusions is increased after splenectomy. Splenectomy should not be delayed when indicated. Preoperative vaccination decreases the chance of post splenectomy infection

Identification of ofloxacin-resistant mycobacterium tuberculosis by PCR-RFLP and sequencing

This study was planned to verify the resistance frequency of Ofloxacin [OFX] against Mycobacterium tuberculosis by polymerase chain reaction restriction fragment length polymorphism [PCR-RFLP] technique and sequencing. Total 366 clinical samples of suspected TB patients were collected from various localities of central Punjab. All of them were found positive by ZN [Zeihl-Nelsen] staining method. Among them, 108 [29.5%] were found negative and 258 [70.5%] positive on PCR based study. The cases not responding to ATT were further characterized by proportion method and by PCR-RFLP to establish the drug resistance. Selected drug resistant case were further sequenced to confirm the results of amplified RFLP. The results showed that out of 118 drug resistant cases, 06 [5.08%], 03 [2.54%] were found resistant to OFX by drug susceptibility testing and PCR-RFLP respectively. The two strains were selected for sequencing procedure. The strain-79 showed point mutation at four points, at codon 70, 71, 76 and 78. The sequence of strain- 81 showed mutation at codon 95.PCR-RFLP is a useful molecular technique for the rapid detection of mutations and may be used to diagnose drug resistance but it should be confirmed by sequencing before starting 2[nd] and 3[rd] generation treatment because the restriction site is the cornerstone of PCR-RFLP and mutation may be occurring elsewhere

Brain drain: doctors' career intentions and associated factors, a questionnaire survey in Pakistan

Objective: To describe Pakistani doctors' career intentions, future practice locations and their satisfaction with work conditions in home country and also, the factors behind leaving and staying in the country

Methodology: This cross sectional study was conducted on 132 junior doctors of POP Hospital and Wah Medical College, Wah Cantt during November-December, 2015. The self-administered questionnaire was designed after extensive literature review and was distributed. Data was analyzed using SPSS version 19 and p value <0.05 was considered statistically significant

Results: Majority of the participants intended to seek postgraduate training in different specialties of medical field [81.8%]. International careers were anticipated by 27.2% of the participants, and UK, Middle East and US was found to be their preferred choices. 72.7% were not satisfied with working conditions in home country. 40.9% reported less likelihood of returning back to Pakistan if opportunity is given to go abroad. Major factors that stood out as reasons for the intention to emigrate included; doctors being paid a high salary, higher educational opportunities, good working conditions abroad and structured career path

Conclusion: Low expected monthly salary, stressful working environment and lack of professional growth were the most important reasons of physician dissatisfaction and migration from developing countries to developed ones

Comparison of clinicopathological characteristics of lymph node positive and lymph node negative breast cancer

Objective: To record various clinicopathological characteristics of breast cancer [BC] in our population and to find an association between these characteristics and axillary nodal metastasis

Methods: This cross-sectional study included 150 BC patients from two tertiary care centers in Karachi from 15[th]February, 2013 to 31[st] March, 2015. Frequencies, percentages, and odds ratio were estimated to find out an association between various clinicopathological characteristics and lymph node status using SPSS version 20

Results: Approximately 75.4% patients had axillary lymph node metastasis ['1-3' IN = 34.4% and '>3' LN =44%]. Menopausal status [p<0.013], tumor grades [II' p <0.03; Mil' p<0.01], and stages [Mil'p <0.002; MV p <0.0001] tumor sizes [T2 p <0.014; T3' p <0.002], perineural invasion [PNI] [p <0.007], lymphovascular invasion [VI] [p <0.0001], and skin and nipple invasion [p <0.024] were significant predictors for '>3' LN metastasis. Association of these variables with '1-3' LN involvement was insignificant

Conclusion: Clinical spectrum of BC remains unchanged in 2016 with most of the patients presenting with high-grade, late-stage advanced disease. Moreover, clinicopathological variables, especially primary tumor size, tumor stage and lymphovascular invasion were significant predictors of >3 lymph node metastasis with high accuracy