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.

Dyschromatosis universalis hereditaria: first case report from Northern Pakistan

Dyschromatosis universalis hereditaria is a rare genodermatosis, first reported from Japan. Later on many cases have been reported from other countries of the world. It is characterized by mottled pigmentation. We report first case of this disorder from Northern Pakistan with positive family history

Frequency and antimicrobial susceptibility pattern of Acinetobacter species isolated from pus and pus swab specimens

To evaluate the frequency and antimicrobial susceptibility pattern of Acinetobacter species isolated from pus and pus swab specimens at a tertiary care setting. Cross-sectional observational study. Department of Microbiology, Armed Forces Institute of Pathology, Rawalpindi, from July 2008 to July 2012. Data regarding positive culture and antimicrobial sensitivity pattern was retrieved from the pus and pus swab culture records of the Microbiology Department, AFIP, Rawalpindi. Only those pus and pus swab specimens which yielded the growth of Acinetobacter species were included in the study. Out of 2781, 1848 were of pure pus while 933 were pus swab specimens. Out of 2538 culture positive isolates, 276 [10.9%] were identified as Acinetobacter species. Among 276 Acinetobacterspp., 245 [88.8%] were Acinetobacter baumannii and 31 [11.2%] were Acinetobacter johnsonii. Male/female ratio of the affected patients was 5.6:1. Doxycycline was the most sensitive antibiotic to which 45% of the tested isolates were sensitive. Sensitivity to all other antimicrobials was 15% or less. About 11% of soft tissue and wound infections are caused by Acinetobacter species in our set up particularly in male. Doxycycline was the most sensitive antibiotic. Sensitivity to all other antimicrobials was 15% or less. In vitro sensitivity to carbapenems is very low

vitro antimicrobial, antioxidant activity and phytochemical screening of Apium graveolens

The present study evaluates the phytochemical screenings, antioxidant activity and antimicrobial assay of Apium graveolens L. The phytochemical screening showed the presence of flavonoids, tannins, saponins and steroids in Apium graveolens while terpenoids was absent. The total phenolic content was slightly higher in methanolic fraction [63.46 +/- 12.00mg GAE/g] followed by ethanol [36.60 +/- 12.28 mg GAE/g] and hexane fractions [34.86 +/- 6.96mg GAE/g]. The flavonoid content was high in methanolic extract [56.95 +/- 7.14mg Quorcetin/g] and low level of the content was found in methylated spirit extract [29.2 +/- 3.15mg Quercitin/g]. Antioxidant activity assayed by FRAP was higher in methanolic fraction [12.48 +/- 1.06 mmole of FeSO[4] equivalent/litre of extract] compared with other extracts. Likewise, good antimicrobial activity was measured by crude ethanol fraction against S. aureus [MIC=0.12 +/- 0.03microg/ml] and S. typhi [MIC= 0.5 +/- 0.2microg/ml]. Results also that ethanolic fraction was effective against A. flavus [MIC= 05 +/- 1.0microg/ml]

Surgical outcome of tethered cord syndrome

Objective: surgical outcome of tethered cord syndrome

Study Design: retrospective descriptive

Period and Place of study: Neurosurgery Department Allied Hospital Faisalabad over 6 months from October 1, 2014 to March 28, 2015

Materials and Methods: forty patients were selected at Neurosurgical Department, Allied Hospital, Faisalabad with age ranging from 2 months to 27 years[average 2.58 years]. These were assessed clinically and their orthopedic and urological aspects were also assessed. MRI of relevant area was advised. Counselling was done regarding surgery under general anesthesia and its outcome. Laminectomy or laminotomy was done. Cord was detethered from its attachments from dura, and, in case of lipomeningocele, from its extradural component. In case of diastometomyelia, bony spur was extirpated and dura- plasty was done. Tight filum terminale was divided. Surgery was done using microsurgery techniques. Postoperative assessment was done immediately at the time of discharge, at 6 and 12 months

Results: a total of 40 patients, M:F ratio was5:3. Average age was 2.58 years.Age was ranging from 2 months to 27 years. Power improved in 44 % to grade 5/5 and 37% in grade 4/5 at one year. There was 65 % improvement in anal sphincters and 71% improvement in urinary sphincters at one year

Conclusion: our clinical and neurological evaluation showed satisfactory outcome after surgery, particularly when performed before the onset of irreversible deficit. Neurological status prior to surgery has profound impact on the outcome

Antimicrobial susceptibility pattern of bacteria isolated from patients with urinary tract infection

To determine the antimicrobial susceptibility pattern of bacterial pathogens in the patients of urinary tract infection reporting at a tertiary care hospital. Laboratory based study. Department of Microbiology, Armed Forces Institute of Pathology, Rawalpindi, from January to December 2012. A total of 440 culture positive bacterial isolates from 1110 urine samples; submitted over a period of one year were included in this study. Identification of bacterial isolates was done by standard biochemical profile of the organisms. The antimicrobial susceptibility of culture positive bacterial isolates was performed by disk diffusion method as recommended by Clinical Laboratory Standard Institute guidelines [CLSI]. Out of the 440 culture positive urine samples, 152 [34.6%] were from indoor patients whereas 288 [65.4%] from outdoor patients. Gram negative bacteria accounted for 414 [94%] of the total isolates while rest of the 26 [6%] were Gram positive bacteria. The most prevalent bacterial isolate was Escherichia [E.] coli 270 [61.3%] followed by Pseudomonas [P.] aeruginosa 52 [12%] and Klebsiella [K.] pneumoniae 42 [9.5%]. The susceptibility pattern of E. coli showed that 96.2% of the bacterial isolates were sensitive to imipenem, 85.1% to amikacin, 80.7% to piperacillin/tazobactam and 72.6% to nitrofurantoin. In case of P. aeruginsosa, 73% bacterial isolates were sensitive to tazobactam/piperacillin, 69.2% to sulbactam/cefoperazone and 65.38% to imipenem. The antibiogram of K. pneumoniae has revealed that 76.1% of the bacterial isolates were sensitive to imipenem and 52.3% to piperacillin/tazobactam. Nitrofurantoin and imipenem were the most effective antimicrobials amongst the Enterococcus spp. as 92.3% showed susceptibility to this bacterial isolate. Majority of the bacterial isolates were sensitive to imipenem and piperacillin/tazobactam while susceptibility to most of the commonly used oral antibiotics was very low. Among the oral antimicrobials, nitrofurantoin showed good susceptibility against Enterobacteriaceae family and Gram positive organisms

In vitro activity of fosfomycin tromethamine against extended spectrum beta-lactamase producing urinary tract bacteria

To determine the in vitro activity of Fosfomycin tromethamine against extended spectrum beta-lactamase producing uropathogens. Experimental study. Department of Microbiology, Armed Forces Institute of Pathology, Rawalpindi, from October 2011 to October 2012. A total of 381 culture positive ESBL producing isolates from 2400 urine samples submitted over a period of one year were included in this study. Identification of isolates was done by standard biochemical profile of the organisms. The antimicrobial susceptibility of culture positive isolates was performed by disk diffusion method as recommended by Clinical Laboratory Standard Institute guidelines [CLSI]. The antimicrobial activity of Fosfomycin to various isolates revealed that 93% of E. coli, 64% Klebsiella spp. 50% Proteus spp. 75% Enterobacter cloacae, 100% Citrobacter freundii, 100% Burkholderia spp. 100% Serratia spp. and 50% Stenotrophomonas maltophilia were susceptible to this chemical compound. Fosfomycin showed excellent effectiveness to most of the common ESBL producing bacteria such as E. coli, Klebsiella and Proteus spp

Useful ethnophytomedicinal recipes of angiosperms used against diabetes in South East Asian Countries [India, Pakistan and Sri Lanka]

This paper is based on data recorded from various literatures pertaining to ethnophytomedicinal recipes used against diabetes in South East Asia [India, Pakistan and Srilanka]

Traditional plant treatments have been used throughout the world for the therapy of diabetes mellitus. In total 419 useful phytorecipes of 270 plant species belonging to 74 Angiospermic families were collected. From the review it was revealed that plants showing hypoglycemic potential mainly belong to the families, Cucurbitaceae [16 spp.], Euphorbiaceae [15 spp.], Caesalpiniaceae and Papilionaceae [13 spp. Each], Moraceae [11 spp], Acanthaceae [10 spp.], Mimosaceae [09 spp], Asteraceae, Malvaceae and Poaceae [08 spp. Each], Hippocrateaceae, Rutaceae and Zingiberaceae [07 spp. Each], Apocynaceae, Asclepiadaceae and Verbenaceae [06 spp. Each], Apiaceae, Convolvulaceae, Lamiaceae, Myrtaceae, Solanaceae [05 spp.each]

The most active plants are Syzigium cumini[14 recipes], Phyllanthus emblica [09 recipes], Centella asiatica and Momordica charantia[08 recipes each], Azadirachta indica [07 recipes], Aegle marmelos, Catharanthus roseus, Ficus benghalensis, Ficus racemosa, Gymnema sylvestre [06 recipes each], Allium cepa, A. sativum, Andrographis paniculata, Curcuma longa [05 recipes each], Citrullus colocynthis, Justicia adhatoda, Nelumbo nucifera, Tinospora cordifolia, Trigonella foenum-graecum, Ziziphus mauritiana and Wattakaka volubilis [4 recipes each]

These traditional recipes include extracts, leaves, powders, tlour, seeds, vegetables, fruits and herbal mixtures

Data inventory consists of botanical name, recipe, vernacular name, English name

Some of the plants of the above data with experimentally confirmed antidiabetic properties have also been recorded. More investigations must be carried out to evaluate the mechanism of action of diabetic medicinal plants

Toxicity of these plants should also be explained. Scientific validation of these recipes may help in discovering new drugs from these medicinal plants for diabetes

Clinical presentation of non-HIV Kaposi sarcoma

Kaposi Sarcoma [KS] is a rare entity. In the north west of Pakistan and Aghanistan, we mostly come across non-HIV related Kaposi sarcoma as Human Immunedeficiency Virus [HIV]. Infections are rare in this part of the world. Here, we present a case of a non-auto Immunedeficiency Disease [AIDS] related KS. A 45-year-old male, Afghan patient presented to our oncology outpatient's unit with multiple subcutaneous nodules. The sites of involvement were the periorbital region, retro-auricular region, forearms, legs, chest and back. Oral mucosa was spared at the nodules. The patient had no visceromegaly at the time of presentation. A biopsy specimen from the retro-auricular region revealed a KS with dermal lymphatic involvement. His serum was negative for the common types of viral infections including Human Immunodeficiency Virus [HIV] on routine serology. His total B-lymphocytes [CD 19+], total T-lymphocyte [CD3+], total CD4+ lymphocyte [CD3+, CD4+] and total CD8+ [CD3+, CD8+] counts were all normal or borderline high. The patient was under treatment with 3 weekly chemotherapeutic regimens of Adriamycin, Bleomycin, Vincristine [ABV] keeping in view socioeconomical constrains, logistical difficulties in getting proper medical care and side effects of other options like radiotherapy for extended surface areas

Bidirectional Glenn shunt without cardiopulmonary bypass

To determine the efficacy of bidirectional Glenn shunt [BDG] without cardiopulmonary bypass [CPB]. Quasi experimental study. The Armed Forces Institute of Cardiology and National Institute of Heart Diseases [AFIC- NIHD], Rawalpindi. Thirty one patients underwent BDG without CPB between January 2006 to December 2007. Subjects for off pump BDG were those who did not require any intracardiac repair, had good sized branch pulmonary arteries, had acceptable PA pressures [< 16 mm Hg], and did not have any significant atrio-ventricular [AV] valve regurgitation. The off pump BDG was performed using veno-venous shunt between the superior vena cava [SVC] and right atrium [RA] following heparinization. All patients underwent discharge echocardiography to assess BDG patency. Statistical significance was determined using t-test with statistical significance at p < 0.05. There were 18 males and 13 females. All patients survived. Twenty seven [87.09%] patients received BDG and 04 patients [12.90%] received bilateral BDG. Atrial septectomy with inflow occlusion was performed in 5 patients. Antegrade pulmonary blood flow was left in 24 [77.41%] of 31 patients. There was significant improvement in postoperative SpO[2] [p = 0.000] in all the cases. There were no postoperative neurologic complications. Sepsis occurred in 2 patients who ultimately recovered. One patient had chylothorax which stopped after three [03] days in ICU. No SVC/PA distortions were noted by discharge echocardiography. Eliminating CPB reduced the cost of the procedure substantially and saved the patients from its inherent complications. BDG without CPB is a safe procedure in selected patients. It avoids CPB related problems and is cost effective, with excellent results

Glycemic status, income, self-monitoring, compliance, education and pattern of pharmacotherapy in 100 patients of type 2 diabetes mellitus

To assess the glycemic status, income, self-monitoring, compliance, education and pattern of pharmacotherapy in 100 type 2 diabetes mellitus patients. Material and This descriptive observational study was carried out at Medical A Unit, Khyber Teaching Hospital Peshawar. A total of 100 type 2 diabetic patients were included through non-probability sampling. Glycosylated hemoglobin of each patient was determined and questions were asked about income, self-monitoring of glycemic status, education and compliance of the patients. Majority of the patients [78%] had poorly controlled diabetic mellitus. Most of the patients [42%] were poor and their monthly income was less than 5000 rupees. Only 57% of the patients monitored their blood sugar once a month. No patient was using glycosylated hemoglobin for assessment of his or her glycemic status. Majority of the patients [82%] were illiterate. Oral hypoglycemic agents were used by 81% of the patients. Most of the patients had poorly controlled diabetes mellitus. Majority of the patients were poor and illiterate and there was poor self-monitoring. Most of the patients were using oral hypoglycemic drugs

Ventricular septal defect in infants and children with increased pulmonary vascular resistance and pulmonary hypertension - surgical management leaving an atrial level communication

To evaluate the surgical and medical efficacy of the patients operated for Ventricular Septal Defect [VSD] with Pulmonary Hypertension and Pulmonary Vascular Resistance [PVR]. Infants and children with elevated PVR and Pulmonary Hypertension are associated with significant mortality and morbidity after surgical closure. Circulatory assist devices and sophisticated medicines may not be available to help in the management of infants and children with elevated Pulmonary artery pressure and resistance. We left Patent Foramen Ovale [PFO] or made atrial communication to decrease the morbidity and mortality associated with the closure of large VSD in this risky group. Sixteen infants and children were operated with median age of 12 months, operated by the same surgeon [IU], from January' 2004 to December' 2005. They were with large VSD of elevated PVR [3.9+0.3] and underwent VSD closure leaving PFO or artificial ASD [5mm]. Surgical approach was through right atrium. Post operatively, all the patients were electively ventilated for 36 hours. They were given intravenous dilators [Glyceral Trinitrate + Phentolamine] and oral Sildenafil up to 1mg /Kg, six hourly. Five cases went into acute pulmonary hypertensive crisis postoperatively, and were rescued by Prostacycline Nebulization. Sixteen patients had VSD as the primary lesion that underwent operation. The overall early mortality was 6.25% [1/16]. There have been no late deaths. Closure of large VSD with elevated PVR can be performed, leaving PFO or artificial ASD, with acceptable mortality and morbidity