Expression analysis of transcription factors in sugarcane during cold stress / Análise de expressão de fatores de transcrição em cana-de-açúcar sob estresse por frio

Transcription factors (TF) are a wide class of genes in plants, and these can regulate the expression of other genes in response to various environmental stresses (biotic and abiotic). In the current study, transcription factor activity in sugarcane was examined during cold stress. Initially, RNA transcript reads of two sugarcane cultivars (ROC22 and GT08-1108) under cold stress were downloaded from SRA NCBI database. The reads were aligned into a reference genome and the differential expression analyses were performed with the R/Bioconductor edgeR package. Based on our analyses in the ROC22 cultivar, 963 TF genes were significantly upregulated under cold stress among a total of 5649 upregulated genes, while 293 TF genes were downregulated among a total of 3,289 downregulated genes. In the GT08-1108 cultivar, 974 TF genes were identified among 5,649 upregulated genes and 283 TF genes were found among 3,289 downregulated genes. Most transcription factors were annotated with GO categories related to protein binding, transcription factor binding, DNA-sequence-specific binding, transcription factor complex, transcription factor activity in RNA polymerase II, the activity of nucleic acid binding transcription factor, transcription corepressor activity, sequence-specific regulatory region, the activity of transcription factor of RNA polymerase II, transcription factor cofactor activity, transcription factor activity from plastid promoter, transcription factor activity from RNA polymerase I promoter, polymerase II and RNA polymerase III. The findings of above results will help to identify differentially expressed transcription factors during cold stress. It also provides a comprehensive analysis of the regulation of the transcription activity of many genes. Therefore, this study provides the molecular basis for improving cold tolerance in sugarcane and other economically important grasses.

Fatores de transcrição (FT) são uma ampla classe de genes em plantas e podem regular a expressão de outros genes em resposta a vários estresses ambientais (estresses bióticos e abióticos). No presente estudo, a atividade do fator de transcrição na cana-de-açúcar foi examinada durante o estresse pelo frio. Inicialmente, as leituras de transcrição de RNA de duas cultivares de cana-de-açúcar (ROC22 e GT08-1108) sob estresse frio foram baixadas do banco de dados SRA NCBI. As leituras foram alinhadas em um genoma de referência e as análises de expressão diferencial foram realizadas com o pacote R / Bioconductor edgeR. Com base em nossas análises no cultivar ROC22, 963 genes TF foram significativamente regulados positivamente sob estresse pelo frio entre um total de 5.649 genes regulados positivamente, enquanto 293 genes TF foram regulados negativamente entre um total de 3.289 genes regulados negativamente. No cultivar GT08-1108, 974 genes TF foram identificados entre 5.649 genes regulados positivamente e 283 genes TF foram encontrados entre 3.289 genes regulados negativamente. Os fatores de transcrição, em sua maioria, foram anotados com categorias GO relacionadas à ligação de proteína, ligação de fator de transcrição, ligação específica de sequência de DNA, complexo de fator de transcrição, atividade de fator de transcrição em RNA polimerase II, atividade de fator de transcrição de ligação de ácido nucleico, atividade de corepressor de transcrição, sequência específica da região reguladora, atividade do fator de transcrição da RNA polimerase II, atividade do cofator do fator de transcrição, atividade do fator de transcrição do promotor do plastídio, atividade do fator de transcrição do promotor da RNA polimerase I, polimerase II e RNA polimerase III. As descobertas dos resultados acima ajudarão a identificar fatores de transcrição expressos diferencialmente durante o estresse pelo frio. Ele também fornece uma análise abrangente da regulação da atividade [...].

Expression analysis of transcription factors in sugarcane during cold stress

Abstract Transcription factors (TF) are a wide class of genes in plants, and these can regulate the expression of other genes in response to various environmental stresses (biotic and abiotic). In the current study, transcription factor activity in sugarcane was examined during cold stress. Initially, RNA transcript reads of two sugarcane cultivars (ROC22 and GT08-1108) under cold stress were downloaded from SRA NCBI database. The reads were aligned into a reference genome and the differential expression analyses were performed with the R/Bioconductor edgeR package. Based on our analyses in the ROC22 cultivar, 963 TF genes were significantly upregulated under cold stress among a total of 5649 upregulated genes, while 293 TF genes were downregulated among a total of 3,289 downregulated genes. In the GT08-1108 cultivar, 974 TF genes were identified among 5,649 upregulated genes and 283 TF genes were found among 3,289 downregulated genes. Most transcription factors were annotated with GO categories related to protein binding, transcription factor binding, DNA-sequence-specific binding, transcription factor complex, transcription factor activity in RNA polymerase II, the activity of nucleic acid binding transcription factor, transcription corepressor activity, sequence-specific regulatory region, the activity of transcription factor of RNA polymerase II, transcription factor cofactor activity, transcription factor activity from plastid promoter, transcription factor activity from RNA polymerase I promoter, polymerase II and RNA polymerase III. The findings of above results will help to identify differentially expressed transcription factors during cold stress. It also provides a comprehensive analysis of the regulation of the transcription activity of many genes. Therefore, this study provides the molecular basis for improving cold tolerance in sugarcane and other economically important grasses.

Resumo Fatores de transcrição (FT) são uma ampla classe de genes em plantas e podem regular a expressão de outros genes em resposta a vários estresses ambientais (estresses bióticos e abióticos). No presente estudo, a atividade do fator de transcrição na cana-de-açúcar foi examinada durante o estresse pelo frio. Inicialmente, as leituras de transcrição de RNA de duas cultivares de cana-de-açúcar (ROC22 e GT08-1108) sob estresse frio foram baixadas do banco de dados SRA NCBI. As leituras foram alinhadas em um genoma de referência e as análises de expressão diferencial foram realizadas com o pacote R / Bioconductor edgeR. Com base em nossas análises no cultivar ROC22, 963 genes TF foram significativamente regulados positivamente sob estresse pelo frio entre um total de 5.649 genes regulados positivamente, enquanto 293 genes TF foram regulados negativamente entre um total de 3.289 genes regulados negativamente. No cultivar GT08-1108, 974 genes TF foram identificados entre 5.649 genes regulados positivamente e 283 genes TF foram encontrados entre 3.289 genes regulados negativamente. Os fatores de transcrição, em sua maioria, foram anotados com categorias GO relacionadas à ligação de proteína, ligação de fator de transcrição, ligação específica de sequência de DNA, complexo de fator de transcrição, atividade de fator de transcrição em RNA polimerase II, atividade de fator de transcrição de ligação de ácido nucleico, atividade de corepressor de transcrição, sequência específica da região reguladora, atividade do fator de transcrição da RNA polimerase II, atividade do cofator do fator de transcrição, atividade do fator de transcrição do promotor do plastídio, atividade do fator de transcrição do promotor da RNA polimerase I, polimerase II e RNA polimerase III. As descobertas dos resultados acima ajudarão a identificar fatores de transcrição expressos diferencialmente durante o estresse pelo frio. Ele também fornece uma análise abrangente da regulação da atividade de transcrição de muitos genes. Portanto, este estudo fornece base molecular para melhorar a tolerância ao frio em cana-de-açúcar e outras gramíneas economicamente importantes.

Partial purification, characterization and application of thermoalkaliphilic proteases from Priestia endophytica, Lysinibacillus cresolivorans and Bacillus subtilis isolated from desert soil

Aims@#Thermophilic proteases are important industrial enzymes because they can be used at high temperatures in various bioprocessing schemes. The bacterial population of the Cholistan desert was explored for thermophilic proteases and their industrial applications.@*Methodology and results@#Three bacterial isolates K1, K5 and K7 were found promising protease producers. These isolates were preliminary identified as Bacillus based on morphological characteristics and biochemical tests (positive for catalase, oxidase and citrate tests, and negative for indole and urease tests). The isolates K1, K5 and K7 were further identified as Priestia endophytica, Lysinibacillus cresolivorans and Bacillus subtilis, respectively by phylogenetic analysis. The isolates grew best at 50 °C and P. endophytica (K1), L. cresolivorans (K5) and B. subtilis (K7) produced larger zones of hydrolysis at 37 °C, 45 °C and 50 °C at pH 7, respectively. The optimum temperature where protease activity was maximum was 65 °C for P. endophytica and L. cresolivorans and 55 °C for B. subtilis, and the optimum pH was 9.@*Conclusion, significance and impact of study@#The proteases produced by these isolates were found active at high temperatures (45 °C to 85 °C) and high pH (9-12), which make them industrially important thermoalkaliphilic proteases. These proteases successfully de-haired cow’s skin and de-stained blood from cotton cloth pieces, which are rarely tested applications of these proteases.

Cashmirins A and B, new antifungal and urease inhibitory prenylated coumarins from Sorbus cashmiriana

Abstract Cashmirins A (1) and B (2), new prenylated coumarins, have been isolated from the EtOAc- soluble fraction of the whole plant of Sorbus cashmiriana Hedlung, Monog. along with seselin (3), scopoletin (4), 3-hydroxyxanthyletin (5) and luteolin (6), reported for the first time from this species. Their structures were elucidated by spectroscopic techniques including MS, 1D and 2D NMR spectroscopy. Both new compounds 1 and 2 were investigated for biological activities and showed significant antifungal and urease inhibitory activities. Compounds 1 and 2 exhibited significant activity against Aspergillus flavus, Macrophomina phaseolina, Trichophyton simii, Trichophyton schoenleinii, and Pseudallescheria boydri. Both compounds also exhibited significant inhibitory activity against Jack bean urease with IC50 values of 28.2±0.12 µM and 30.3±0.18 µM, respectively compared to thiourea used as positive control.

Intestinal spirocheteosis in a patient with celiac disease

A young girl presented to us with recurrent diarrhea along with a history of 5 kg weight loss in one year. On examination, she appeared pale, while her laboratory reports showed a low hemoglobin, mean corpuscular volume [MCV] and serum albumin. Her erythrocyte sedimentation rate [ESR] was slightly raised with her iron profile suggestive of iron deficiency anemia. Viral markers, human immunodeficiency virus [HIV] serology along with thyroid profile were all unremarkable. There was no history of tuberculosis, and purified protein derivative [PPD] skin test was also negative. Computed tomography [CT] abdomen showed thickening of the terminal ileum with multiple enlarged lymph nodes. An esophagogastroduodenoscopy [EGD] along with colonoscopy was done. Multiple biopsies were taken, which were suggestive of sprue along with intestinal spirochetosis. Her tissue transglutaminase [TTG] was negative while deamidated gliadin peptide [DGP] was positive. She was kept on gluten-free diet and started on tablet metronidazole. This case shows that intestinal spirochetosis should be kept in mind in patients belonging to lower socio-economic status, who present with chronic diarrhea symptoms

Axillary artery aneurysm

Axillary artery aneurysm is a rare condition. Though mostly asymptomatic, it should be treated early as it can develop thrombosis, distal embolisation, pressure symptoms or rupture. We report here a case of a 24-year woman who presented with neurological deficit in her arm because of pressure by axillary artery aneurysm. Standard treatment of these aneurysms is excision and replacement with an interposition graft. We used reversed saphenous vein as interposition graft between axillary and brachial artery

Hepatic Sarcodosis presenting as portal hypertension in a young boy

A 13-year-old boy, known case renal stone disease came with the complaints of abdominal pain along with low grade fever. On examination, hepatosplenomegaly was noted while his lab reports showed a low hemoglobulin with a raised ESR. His blood and urine cultures showed no growth. Viral markers, autoimmune profile, C and p ANCA were all negative apart from a raised serum IgG level. Ultrasound abdomen showed a hyperechoic liver with an enlarged spleen along with splenic varices and minimum ascites. Ultrasound hepatic doppler was normal. Serum AFP levels were normal while workup for Wilson's disease was negative. Fibroscan showed F4 fibosis. CT scan abdomen showed an enlarged left lobe of the liver along with an enlarged spleen. His EGD revealed varices. So liver biopsy was done that was suggestive of chronic granulomatous disease with ZN stain testing negative for TB.PPD, urine for AFB were both negative. Serum ACE levels were raised. He started ATT therapy but his condition did not improve. So, on the suspicion of hepatic sarcoidosis, he started on steroids and had a drastic improvement in his condition

use of Karnofsky Performance Status [KPS] as a predictor of 3 month post discharge mortality in cirrhotic patients

Aim: Is Karnofsky Performance Status [KPS] a predictor of 3 month post discharge mortality in cirrhotic patients?

Background: Cirrhotic patients often experience an abrupt decline in their health, which often leads to frequent hospitalization and can cause morbidity and mortality. Various models are currently used to predict mortality in cirrhotics however these have their limitations. The Karnofsky Performance Status [KPS] being one of the oldest performance status scales, is a health care provider - administered assessment that has been validated to predict mortality across the elderly and in the chronic disease populations

Methods: We used the KPS performance status scale to envisage short-term mortality in cirrhotic and HCC patients who survive to be discharged from hospital

Results: Our study showed that KPS one week post-discharge, child pugh score, hospital stay, international normalized ratio, serum albumin, total bilirubin and serum creatinine showed statistical significance on univariate analysis. On multivariate analysis, KPS was found to be statistical significant predictor of 3-month mortality

Conclusion: Hence KPS can be utilized to identify cirrhotic patients at risk of 3-month post discharge mortality

Disease patterns among hajj pilgrims attending medical facilities of pakistan Hajj medical mission 2016 [1437 hijri]

Objective: The purpose of this study was to determine the frequency of different diseases among Pakistani Hajj pilgrims attending the medical facilities of Pakistan Hajj Medical Mission during Hajj 2016 [1437 hijri]. Study Design: A descriptive cross sectional study. Place and Duration of Study: Pakistan Hajj Medical Mission hospitals and dispensaries at Makkah Mukkarrmah, Medina Munawwarah and Jeddah, from 10[th] Aug 2016 to 5[th] Oct 2016

Material and Methods: All Pakistani Hajj pilgrims reporting to various Pakistan Hajj Medical Mission Hospitals and dispensaries at Makkah, Medina and Jeddah were included in the study. A universal sample of all patients reporting to the Hajj medical mission [A total of 184,496 OPD visits] was used. The patient were initially assessed in Emergency Reception[ER] by medical officers and then referred to respective specialists if required. A specially designed proforma having information regarding name, age, disease and its duration was prepared and filled for each patient separately. A second hospital/dispensary visit of the Hajj pilgrim was considered separately on a new proforma. SPSS version 17 was used to analyze the data

Results: There were a total of 184,496 OPD [Out Patient Department] visits by Pakistani Hajj pilgrims during the study period. Age of the patients ranged from 20-96 years, 74.5% [n=137, 449] were male and 25.5% [n=47047] were female. Number of patients suffering from various diseases and their percentage in order of frequency was; respiratory diseases 29% [n=53187], musculoskeletal disorders 18% [n=33838], gastrointestinal diseases 15% [n=26696], Ear Nose and Throat [ENT] diseases 8% [n=14448], skin disorders 6% [n=10937], eye disease 3% [n=4530], mouth and dental diseases 3% [n=6101], wounds, fractures and burns 3% [n=6186], cardiovascular diseases 2% [n=4433], gynecological disorders 2% [n=4357], infectious disease 1% [n=1055], minor surgeries <1% [n=620], psychiatric disorders <1% [n=40] and other miscellaneous complaints 5% [n=9889]

Conclusion: Respiratory illness was the commonest disease among Pakistani Hajj Pilgrims while musculoskeletal and gastrointestinal complaints were also high. This study helps to identify the common diseases encountered during Hajj Medical Mission and may aid in the better preparedness of such missions in future

Evaluation of different formulations of IGRs against Aedes albopictus and Culex quinquefasciatus (Diptera: Culicidae)

Objective: To test the relative efficacy of pyriproxyfen and methoprene on mortality, deformity, inhibition and emergence to adult stages of Culex quinquefasciatus and Aedes albopictus. Methods: Serial dilutions (0.01-0.05 mg/L) of methoprene, pyriproxyfen 0.5 water dispersible granules (WDG) and pyriproxyfen 1.0 WDG were used to assess mortality and inhibition of 3rd instar larvae of Aedes albopictus and Culex quinquefasciatus. Each concentration and control was replicated four times in completely randomized design. Data on larval mortality, growth inhibition, deformities and adult's emergence was recorded weekly. On the basis of best comparative performance, the efficacy of pyriproxyfen 1.0 WDG at 0.1 g/m

Evaluation of different formulations of IGRs against Aedes albopictus and Culex quinquefasciatus (Diptera:Culicidae)

Objective: To test the relative efficacy of pyriproxyfen and methoprene on mortality, deformity, inhibition and emergence to adult stages of Culex quinquefasciatus and Aedes albopictus. Methods: Serial dilutions (0.01–0.05 mg/L) of methoprene, pyriproxyfen 0.5 water dispersible granules (WDG) and pyriproxyfen 1.0 WDG were used to assess mortality and inhibition of 3rd instar larvae of Aedes albopictus and Culex quinquefasciatus. Each concentration and control was replicated four times in completely randomized design. Data on larval mortality, growth inhibition, deformities and adult's emergence was recorded weekly. On the basis of best comparative performance, the efficacy of pyr-iproxyfen 1.0 WDG at 0.1 g/m3 was also tested in the field by collecting treated water samples monthly for 1–6 months after field application. Twenty five 3rd instar larvae of Aedes and Culex spp. of the same cohorts were used for bioassays and compared with larvae in control cups containing 1 L of untreated tap water. Results: Results revealed variations in fatality of different insect growth regulators (IGRs) to the 3rd instar larvae of Culex and Aedes mosquitoes. Among the IGRs, pyr-iproxyfen 1.0 WDG was found best that exhibited significantly high emergence inhibition against Culex and Aedes spp. Based on the results, the IGRs were classified in terms of the tested parameters in order of pyriproxyfen 1.0 WDG > pyriproxyfen 0.5 WDG > methoprene. In case of field studies, pyriproxyfen 1.0 WDG, pool data of the entire target treated sites showed minimum adult emergence from water sampled of habitats treated with 0.1 g/m3 of pyriproxyfen 1.0 WDG. Conclusions: It is thus concluded that IGRs can be utilized as environment friendly control measures for Culex and Aedes spp. of mosquitoes on small and large scale. This will reduce the use of conventional insecticides by the public health authorities and help in reducing selection pressure of insecticides.

Hypercholesterolemia; comparison of the efficacy of simvastatin 20mg with atorvastatin 20mg in lowering low density lipoprotein in patients with hypercholesterolemia

To compare the efficacy of Simvastatin with Atorvastatin in lowering Low Density Lipoprotein Cholesterol [LDL-C] in patients with Hypercholesterolemia in a tertiary care hospital. Prospective, observational, single center study. Department of Medicine, Khyber Teaching Hospital, Peshawar. December 2011 to December 2012. A total of 200 cases having base line fasting LDL-C level of >/= 130 mg/dl and meeting the inclusion criteria were included in the study through both outpatient department [OPD] and admitted patient. After detailed history and clinical examination, all patients were divided randomly into two groups, A and B. Patients in Group A were given Simvastatin 20mg/day and Group B received Atorvastatin 20mg/day. Fasting blood samples were taken from the selected patients in the start of study and after 12 weeks. A total of 200 subjects with a serum LDL-C level >/= 130 mg/dL were included in the study. They were divided into 2 groups randomly, 100 in each group. Each group comprised of 75 male and 25 female. The mean age in group A was 52 years and in group B it was 54 years. The age ranges between 40 years and 73 years. Mean base LDL-C level was 165 mg/dl in group A and 170mg/dl in group B. Simvastatin 20 mg/dl reduced LDL-C level by 26% and Atorvastatin 20mg/dl reduced LDL-C level by 33% after 12 weeks of treatment. Atorvastatin is a more effective drug to reduce serum LDL cholesterol than Simvastatin in the same doses

Falciparum malaria; its diverse clinical spectrum: a descriptive study of 150 admitted patients

To determine how Falciparum Malaria can present clinically in various ways. A study conducted on 150 admitted patients. This descriptive case-series, single center study was conducted in Medical B Unit, Khyber Teaching Hospital, Peshawar from November 2012 to June 2013. A total of 150 patients admitted with falciparum malaria were studied. A detailed history and clinical examination was performed. The falciparum malaria was diagnosed by examining peripheral blood film. All patients were thoroughly investigated to find out the complications of falciparum malaria. Out of 150 patients, 95 [63.33%] were males 55 [36.67%] were females with mean age of 30 years. Fever was present in all patients, rigors and chills in 93%, headache and vomiting in 60%, jaundice in 50%, confusion in 37%, abdominal pain in 26% were main presentations. Other presentations were Oliguria, hypotension, cerebral malaria, dyspnea and cough, hypoglycemia and seizures. Clinical examination showed splenomegaly [58%], hepatomegaly [46%], hepato-splenomegaly [22%] of patients. Laboratory investigation showed Hemoglobin < 10gm% in 22% and leukocytosis > 12000/?l in 10% of patients. As Falciparum Malaria presents in multiple ways clinically, it should be considered as a possibility in all febrile illnesses

Stability Indicating Simultaneous Equation Method for Determination of Domperidone and (S)-Esomeprazole Magnesium in Capsule Dosage Form Using UV-Spectrophotometer.

Aims: Stability indicating simultaneous equation method for determination of Domperidone and Esomeprazole Magnesium in capsule dosage form using UVSpectrophotometry. Study Design: A new simultaneous equation method was developed and validated for the determination of esomeprazole magnesium and domperidone in capsule dosage form. Place and Duration of Study: Department of Pharmaceutical Chemistry, Invertis Institute of Pharmacy, Invertis University, Bareilly, Uttar Pradesh during July 2012 to June 2013. Methodology: Simultaneous equation method was performed for estimation of dosage form and degradants. Results: The maximum wavelength (λmax) was found to be 299 nm for esomeprazole magnesium and 287 nm for domperidone. The linearity range was found to be 1-6 μg ml-1 (r2= 0.998) and 5-30 μg ml-1 (r2= 0.999) for esomeprazole magnesium and domperidone, respectively. The value of limit of detection and limit of quantification was 0.116 and 0.386 μgml-1 for esomeprazole magnesium and 0.657 and 2.18 μgml-1 for domperidone, respectively. Forced degradations were carried out under acid, base, thermal, photolytic and oxidative stress conditions. The method was satisfactorily validated as per the ICH guideline. Conclusion: This study shows that the proposed spectrophotometric method is useful for the routine determination of esomeprazole magnesium and domperidone in its combined pharmaceutical dosage form.

Teenage jejunoileal diverticulosis

Presentation of jeujunoileal diverticulosis in young age is virtually unknown. It is associated with middle or old age. It is usually asymptomatic but may present with vague abdominal pain and episodic nausea, vomiting or diarrhoea. It can lead to complications like bleeding, perforation and obstruction. We had 3 cases of jejuno-ileal diverticulosis presenting in their teens. They had resection and anastomosis resulting in improved health

Clinicopathological study of cutaneous chromoblastomycosis in Pakistan

To recognize commonest clinical presentations of chromoblastomycosis and isolate fungi responsible for causing chromoblastomycosis in our part of the world. The study included all patients with clinical diagnosis of chromoblastomycosis and skin histopathology and fungal culture confirming clinical diagnosis. The disease started in all our patients during their childhood. All presented with psoriasiform plaques and showed infection with Phialophora dermatitidis Psoriasiform plaques can be the clinical presentation of chromoblastomycosis. P. dermatitidis is the fungus commonly seen in our enrolled patients

Spontaneous bacterial peritonitis. Comparison of treatment with ciprofloxacin and ceftriaxone in patients with cirrhosis liver and ascites

To compare the efficacy of ciprofloxacin with ceftriaxone in the treatment of spontaneous bacterial peritonitis in patients with cirrhosis liver and ascites. This hospital based quasi-experimental study. Department of Medicine, Khyber Teaching Hospital Peshawar. October, 2009 to April, 2010. A total of 200 patients were selected by non-probability purposive sampling method after obtaining an informed consent. Sample size was calculated through WHOS statistical calculator. All the selected patients had clinical and biochemical evidence of cirrhosis liver and spontaneous bacterial peritonitis. Both sexes were included in the selected patients had clinical and biochemical evidence of cirrhosis liver and spontaneous bacterial peritonitis. Both sexes were included in the study. They were randomly divided into two groups; group I was treated with ciprofloxacin and group II was treated with ceftriaxone. 200 patients including 124 males and 76 females with spontaneous bacterial peritonitis were included in the study. 100 patients each were treated with ciprofloxacin and ceftriaxone in the two groups. 82% responded favourably to 5 days course of I/V 200 mg ciprofloxacin and 91% were cured with 5 days therapy of I/V 2gm ceftriaxone. Both ceftriaxone and ciprofloxacin are equally effective in the treatment of spontaneous bacterial peritonitis

Double flap patch closure of VSD with elevated pulmonary vascular resistance: an experience at AFIC/NIHD

To determine the 30 days outcome measured in terms of morbidity and mortality in cases of ventricular septal defect [VSD] with increased pulmonary vascular resistance [PVR] managed with double flap patch closure. Case series. Armed Forces Institute of Cardiology [AFIC/NIHD], Rawalpindi, from December 2005 to December 2008. Forty patients with VSD having PVR 9.58 + 4.33 wood units underwent double flap patch closure. The patch was fenestrated as one half of the expected aortic annulus diameter. A separate flap patch 5 mm larger than fenestration was attached to superior upper one third margins of fenestration. The patch was placed with flap to open towards the left ventricular apex. Modified ultra filtration [MUF] was employed in every case and sildenafil was given postoperatively. The age of patients ranged from 1 to 28 years with a mean of 6.66 + 5.70 years. There were 22 males and 18 females. All patients were weaned off from inotropic and ventilatory support as earlier as possible postoperatively with intensive care unit [ICU] stay of 77.15 + 54.56 hours. Postoperative pulmonary artery pressures were reduced to 42.63 + 10.86 mmHg as compared to pre-operative pulmonary artery pressures of 88.3 + 15.2 mmHg. Postoperatively 11 patients with suprasystemic pulmonary artery pressures and desaturation went into pulmonary hypertensive crisis in which immediate 2D echo evidenced the functioning flap valve with right to left shunt. There was only one death [early] out of 40 patients with an overall mortality of 2.5% along with limited morbidity. Double flap patch is an inexpensive, easy to construct technique with low morbidity and mortality in cases of VSD with raised PVR

Helicobacter pylori detection in chronic gastritis: a comparison of staining methods

Background: Helicobacter pylori is an important cause of chronic gastritis, gastric ulceration and gastric malignancies as gastric carcinoma and MALT lymphoma. Its definitive diagnosis is based on histopathology. Routine H and E stain is not very effective in its detection, immune-stains and fluorescent stains are costly. Need for simple cheap and sensitive stain has always been a topic of hot debate and extensive research. Method: paraffin embedded blocks of all adult patients diagnosed as chronic gastritis/gastric ulceration with no accompanying gastric pathology as hypertrophic gastropathys, and neoplasias were taken into study. Three sections of 4 micron were cut and stained with routine H and E, Giemsa, and Cresyl fast violet. Results: Total number of patients was 50. Out of these 37 [74%] were males and 13 [26%] were females. Mean age of the patients was 50.4 years. Thirty-four percent [34%] were positive in normal H and E stain, 68% were positive in Giemsa and 76% were positive in Cresyl fast violet. Conclusion: Cresyl fast violet is a good stain for diagnosis of H. pylori gastritis. Keywords: H pylori, chronic gastritis, H pylori staining methods

Experience of Fontan surgery at Armed Forces Institute of Cardiology /National Institute of Heart Diseases [AFIC-NIHD]

To assess the overall outcome and success of Fontan surgery at our institute. AFIC-NIHD Rawalpindi. 01 September 2005 to 31 March 2010. For this retrospective study, institute's cardiac surgery database was used. Patients of single ventricle physiology, who had normal Left Ventricular End-diastolic Pressure and pulmonary artery pressures, were included. Data was analyzed using SPSS version 16. A total of 34 Fontan procedures were done. The mean age at operation was 4.83 +/- 1.37 years. There were 22 [64.7%] males and 12 [35.3%] females. Twenty five [73.5%] had a staged Fontan [successful previous Bidirectional Glenn's shunt, BDG]. Nine [26.5%] were primary Fontan procedures [no successful previous Bidirectional Glenn's shunt, BDG]. Thirty two [94.8%] were Extra Cardiac Conduit Fontan [ECCF] and 2[5.8%] were Intra Cardiac Fontan. Mean Bypass time was 132.65 +/- 48.44 minutes. Aorta was cross clamped in intracardiac Fontan and its mean time was 43.31 +/- 5.85minutes. Fenestration was employed in 14 [41.2%] patients. Mean pre-operative oxygen saturations were 77.41 +/- 10.27%, which significantly increased to 93.94 +/- 3.96% post-operatively [p<0.001]. In-hospital mortality was 2 [5.8%]. Fontan surgery has acceptable morbidity and mortality in our set up