Utilization of Integrative Technique for Partial Recovery of Proteases from Soil Microbes

Aqueous two-phase system (ATPS) is an efficient, cost effective, fast, simple and ecofriendly method for the recovery of biomolecules. In the present study, an ATPS composed of polyethylene glycol and ammonium sulphate (NH4)2SO4 was used for the partial purification of proteases from microbial source. The effects of different parameters such as molecular weight of PEG (4000, 6000 and 10000), concentration of PEG (15, 17.5 and 20 %) and concentration of (NH4)2SO4 (7.5, 8.3, 9.1 and 9.9 %) on the partitioning behavior of proteases at room temperature were investigated. Generally, increasing the concentration of PEG and (NH4)2SO4 moved the protease to the top i.e., polymer-rich phase. Increasing the molecular weight of PEG from 4000 to 10000 the partition coefficient decreased subsequently. The highest partition coefficient i.e., 3.32 and maximum activity i.e., 16.06 soxhlet unit was found in an optimum system composed of 20 % PEG 4000 and 9.9 % (NH4)2SO4

Diagnostic yield of transthoracic echocardiography for stroke patients in a developing country

OBJECTIVE: To assess the utility and cost effectiveness of transthoracic echocardiography (TTE) as part of stroke workup in a developing country. METHODS: All patients over the age of 14 years with acute stroke were prospectively enrolled in Aga Khan University stroke databank from August 1999 to May 2001 (22 month period). All patients were evaluated by a consultant neurologist and underwent standard stroke work up such as neuroimaging, TTE, carotid doppler sonography and blood tests e.g. serum glucose, lipid profile, blood urea nitrogen and serum creatinine. For the purpose of this study, patients with ischemic stroke were identified and their echocardiographic data was retrieved and analyzed. TOAST criteria was used to identify findings indicating cardioembolic stroke. RESULTS: Ischaemic stroke was identified in 393 (76%) patients. Of these 278 (71%) patients underwent transthoracic echocardiography as part of stroke workup. Although 199 (73%) patients had abnormal echocardiogram; only 43 (15.5%) had findings suggestive of possible cardioembolism as defined by the TOAST criteria. CONCLUSION: Tranthoracic echocardiography is a valid diagnostic modality for stroke workup. However there is a need to further investigate the specific indications which can be used to prioritize patients for undergoing echocardiography to minimize cost

PO10.19 clinical spectrum of pseudo seizures at a tertiary care hospital Karachi: a video EEG based study

Background: Nonepileptic seizures are involuntary episodes of movement, sensation, or behaviors (e.g., vocalization, crying and other expressions of emotion), that do not result from abnormal cortical discharges. The events typically occur in teenage patients with anxiety and affective disorders. Psychotherapy, behavioral therapy, drugs, hypnosis and placebo are mainstay of treatment. There is no published data on clinical characteristics of pseudo-seizures from Pakistan. The objective of our study is to describe the clinical characteristics of pseudoseizures in a tertiary care hospital in Karachi, Pakistan. Methods: This is a descriptive study. Patients with pseudo-seizures were identified from departmental Video EEG monitoring register, over a period of three years (2004 2006). Their neurophysiologic and video recording data was reviewed and their demographic, clinical and neurophysiologic data was recorded and analyzed. SPSS version 15.0 was used for analysis. Results: A total of 132 patients underwent video EEG during the period and 51 (38%) had pseudo-seizures. Their mean age was 26.7±15.3 years. Thirty-two (63%) were female and 19 (37%) were male. Twelve (23.5%) patients were known epileptics. Median recording time was 24 (range; 0.33 55) hours and median number of events was 3 (range; 1 14). Limb movements were most common manifestation (67%) followed by muteness with no response to verbal commands (49%), behavioral symptoms (35.5%), ocular findings i.e. eye blinking and closure (25.5%), depressed conscious level (13.7%), headache (10%) and GI symptoms (10%). Six (11.8%) patients experienced epileptic seizures, in addition to pseudo-seizures. Conclusions: Pseudo-seizures are common in patients referred for video EEG monitoring. It is more common in young women. Limb movements, muteness and behavioral symptoms are most common manifestations. The pseudo-seizures and epileptic seizures may coexist in the same patient

Heterologous Expression of Genes in Plants for Abiotic Stresses

Abiotic stresses are considered to be the major factors causing a decrease in crop yield globally, these stresses include high and low temperature, salinity, drought, and light stress etc. To overcome the consistent food demand for the ever-growing population, various genes from micro-organisms and non-plant sources have been expressed in transgenic plants to improve their tolerance against abiotic stresses. Gene expression in transgenic plants through conventional methods are time-consuming and laborious that’s why advanced genetic engineering methods for example Agrobacterium-mediated transformation and biolistic methods are more accurate, useful, and less time-consuming. This review provides an insight into various bacterial genes for example mtID, codA, betA, ADH, IPT, DRNF1 and ggpPS, etc. that have been successfully expressed in transgenic plants against various abiotic stress for stress tolerance enhancement and crop yield improvement which exhibited good encouraging results. Genes from yeast (Saccharomyces cerevisiae) have been introduced in transgenic plants against drought and salinity stress. All these genes expressed from non-plant sources in plants can be very helpful to enhance crops for better yield productivity in the future to meet the demands of the consistently rising population of the world

Frequency of Gγ-globin promoter −158 (C>T) XmnI polymorphism in patients with homozygous/compound heterozygous beta thalassaemia

BackgroundResponse to hydroxyurea therapy in homozygous or compound heterozygous beta thalassaemia (BT) has been reported as more favourable in the presence of XmnI polymorphism. The prevalence of XmnI polymorphism may vary with BT phenotypes and genotypes, and differs geographically in distribution. Prevalence of XmnI polymorphism is not known in northern Pakistan.ObjectiveTo determine the frequency of Gγ-globin promoter −158 (C>T) XmnI polymorphism (XmnI polymorphism) in patients with homozygous or compound heterozygous beta thalassaemia.MaterialsPolymerase chain reaction (PCR) for common beta thalassaemia mutations and Gγ-globin promoter −158 (C>T) XmnI polymorphism was performed on 107 blood samples of transfusion dependent beta thalassaemia (BT) patients in Pakistan. One hundred samples of unrelated BT traits and 94 samples of healthy subjects as controls were also analysed for BT mutations and XmnI polymorphism.ResultsOut of 301 DNA samples, XmnI polymorphism was detected in 71(24%); in normal controls, XmnI polymorphism was detected in 34/94 (36%) subjects; while in homozygous/compound heterozygous BT, it was detected in 14/107(13%) patients (Fisher’s exact test, p=.0002). In heterozygous BT group, XmnI polymorphism was detected in 23/100 subjects (Fisher’s exact test, p=.03 with normal controls, and p=.049 with homozygous/compound heterozygous BT). The most common BT genotype was Frame Shift (Fr) 8–9/Fr 8–9, and none of the patients with this genotype had XmnI polymorphism. The second most common genotype was IVSI-5/IVSI-5; 4/26 (15%). Cases with this genotype had XmnI polymorphism.ConclusionXmnI polymorphism in homozygous/compound heterozygous BT group is 13%. The most common genotype associated with XmnI polymorphism was IVSI-5/IVSI-5

Neighbourhood oriented TDMA scheme for the internet of things-enabled remote sensing

Throughout the world, Internet of Things (IoT) have been used in different application areas to assist human beings in numerous activities such as smart buildings and cities via remote sensing-enabled techniques. However, simultaneous transmission of packet(s) by multiple devices Ci, which are interested to start a communication session with a common receiver device, is one of the challenging issues associated with these networks. In the literature, various mechanisms have been presented to resolve the aforementioned issue without changing the technological infrastructures; however, neighbourhood information of sensor nodes is not considered yet. In IoT-enabled remote sensing, neighbourhood information of various devices plays a vital role in developing a reliable communication mechanism specifically for scenarios where multiple devices Ci are interested to start communication with a common destination module. In this paper, a neighbourhood-enabled TDMA scheme is presented for the IoT to ensure the concurrent communication of multiple devices Ci with a common destination device Sj preferably with a minimum possible packet collision ratio (if avoidance is not possible). The proposed scheme bounds each and every member device Ci to assign a dedicated time slot to its neighbouring devices in the operational IoT network. Furthermore, neighbouring devices Ci are forced to communicate within the assigned time slot. Simulation results have verified that the proposed scheme is ideal solution compared to the existing schemes for the IoT and other resource-limited networks particularly in scenarios where the deployment process is random

Diagnosing Beta Thalassemia trait in a developing country

BackgroundBeta Thalassemia trait (BTT) is diagnosed by detecting hemoglobin A2 (HbA2) >3.8% on either High Performance Liquid Chromatography (HPLC) or cellulose acetate electrophoresis (CAE). HPLC is an accurate and reproducible but costly alternative to more conventional CAE which is labor intensive but easy to interpret and inexpensive.ObjectiveTo determine the sensitivity of CAE and HPLC keeping PCR as gold standard for the diagnosis of BTT.Study DesignCross sectional.Place and Duration of StudyArmed Forces Institute of Pathology Rawalpindi. May 2014 to January 2015.Patient and MethodsFiveml EDTA anti-coagulated blood was collected from 100 PCR proven cases of BTT. HbA2 levels were measured by running samples directly on HPLC. But for CAE, first a hemolysate was prepared which was then applied to cellulose acetate membrane at an alkaline pH (7.9). After elution of HbA2 band in Tris EDTA borate buffer (pH of 8.9), HbA2 concentration was calculated by measuring its absorbance in a photometer at a wavelength of 416nm.ResultsMean age of the patients was 28.8±8.1 year. The most common mutation was Fr 8–9 (35%) followed by IVS1-5 (25%) mutation. Mean HbA2 levels by CAE and HPLC were 4.97±0.42 and 5.54±0.59 respectively. All the patients had HbA2>4% on both CAE and HPLC. None of our patients had false negative result either on CAE or HPLC.ConclusionCAE has comparable sensitivity with HPLC for detection of Beta Thalassemia Trait

Surgical Outcome of Otogenic Brain Abscess in Chronic Suppurative Otitis Media

Objective: To determine the surgical out come of otogenic brain abscess in chronic suppurative otitis media.Material and Methods: This descriptive study was conducted at the departments of ENT, Head, Neck Surgery and Neurosurgery Postgraduate Medical Institute (PGMI) Lady Reading Hospital Peshawar from June 2004 to May 2009 with a total duration of five years. The diagnostic criteria were detailed history, thorough Neuro-otological examination and radiological investigations. These patients were admitted into ENT Department and they were put on quadruple therapy. CT scans were performed on emergency basis and as the patients were diagnosed as having otogenic brain abscess urgent evacuation of abscess was done by neurosurgeon. All the patients with established brain abscess were included in this study while those patients having small multiple abscesses, early cereberitis and non otogenic brain abscesses were excluded from study. A well informed consent was taken from patient explaining the procedure, its risks, benefits and associated complications. The abscesses were primarily treated by neurosurgeons and the ears were later on treated by ENT surgeons. There was neither intra operative nor postoperative complication.Results: This study included 22 patients in the age range of 12-51 years with mean age 25 years. Sixteen were male and six were female, with female to male ratio 1 : 2.6. These patients presented with history of discharging ear, headache, vomiting and disorientation. The signs of disease in these patients were granulation tissues/ cholesteatoma in ear and papilloedema. CT scans were performed in all 22 cases (100%) while MRI in 10 cases (45.45%). There were single brain abscess in 14 cases (63.63%) while 8 cases (44.45%) having multiple brain abscesses. Mainly abscess were found in temporal region followed by cerebellar. The patients with chronic ear disease were treated by radical mastoidectomy and modified radical mastoidectomy after treating the brain abscess by Neurosurgeon. The Neurosurgical procedures performed were capsulotmies in 13 cases (59.09%), Bur hole aspiration in 7 cases (31.81%) and Trans mastoid excision in 2 cases (9.09%). These patients were followed for 3 monthly having CT scan with contrast with bone window.Conclusion: We concluded from our study that otogenic brain abscess is a life threatening complication, so early identification with help of CT scan and prompt surgical intervention with appropriate treatment of middle ear (primary disease) is essential for better outcom

Novel VPS13B Mutations in Three Large Pakistani Cohen Syndrome Families Suggests a Baloch Variant with Autistic-Like Features.

BackgroundCohen Syndrome (COH1) is a rare autosomal recessive disorder, principally identified by ocular, neural and muscular deficits. We identified three large consanguineous Pakistani families with intellectual disability and in some cases with autistic traits.MethodsClinical assessments were performed in order to allow comparison of clinical features with other VPS13B mutations. Homozygosity mapping followed by whole exome sequencing and Sanger sequencing strategies were used to identify disease-related mutations.ResultsWe identified two novel homozygous deletion mutations in VPS13B, firstly a 1 bp deletion, NM_017890.4:c.6879delT; p.Phe2293Leufs*24, and secondly a deletion of exons 37-40, which co-segregate with affected status. In addition to COH1-related traits, autistic features were reported in a number of family members, contrasting with the "friendly" demeanour often associated with COH1. The c.6879delT mutation is present in two families from different regions of the country, but both from the Baloch sub-ethnic group, and with a shared haplotype, indicating a founder effect among the Baloch population.ConclusionWe suspect that the c.6879delT mutation may be a common cause of COH1 and similar phenotypes among the Baloch population. Additionally, most of the individuals with the c.6879delT mutation in these two families also present with autistic like traits, and suggests that this variant may lead to a distinct autistic-like COH1 subgroup