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Background: Enhancer of zeste homolog 2 (EZH2) is one of the major epigenetic modifiers involved in the transcriptional repression of target genes through trimethylation of H3K27 (lysine 27 residue of histone H3). Deregulated expression of both EZH2 and H3K27me3 has been implicated in the biological behavior and prognostic outcome of various malignancies. Aim: To assess the role of EZH2 and H3K27me3 in the carcinogenesis of urothelial carcinoma of urinary bladder. Materials and Methods: One hundred fifty consecutive urothelial carcinoma cases of urinary bladder (54.7% high-grade) were included in this study. Immunohistochemical analysis for EZH2 and H3K27me3 was performed on whole tissue sections. A multiplication score obtained by multiplying staining intensity and proportion of positively stained neoplastic cells was used for assessment. Results: EZH2 showed a significant correlation with the tumor grade and lamina propria invasion (p < 0.001). The cases with high EZH2 expression showed a significantly high proliferative index (Mean- 32.7%; p < 0.001). In contrast, negative and low expression of H3K27me3 was significantly more common in high-grade cases (p = 0.006). The expression of H3K27me3 was significantly associated with lamina propria (p = 0.01) and deep muscle invasion (p = 0.007). EZH2 showed a significantly higher expression in the high-grade invasive areas as compared to the high-grade non-invasive areas of the same tumor (p = 0.03). Conclusions: This study establishes an important role of the key epigenetic regulators EZH2 and H3K27me3 in the pathobiology of urothelial carcinomas. Strong expression of EZH2 and weak expression of H3K27me3 are associated with higher grade, proliferative index and invasive behavior.
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Background: Chronic cardiovascular morbidities are major public health concern. The objective was to as-sess awareness and practice of selfcare activities among patients with chronic cardiovascular morbidity in Burdwan Medical College (BMC) and to explore constraints in selfcare practice. Materials & Methods: A facility-based, explanatory sequential, mixed-method study was conducted in Cardi-ology super-specialty outpatient department of BMC, West Bengal. A calculated sample of 185 patients with chronic cardiovascular morbidity were selected randomly; interviewed with predesigned, pretested schedule (adapted from H-SCALE) for quantitative component. For qualitative component, eight study participants, se-lected purposively, were interviewed with In-depth-interview guide. Multivariable logistic regression was done to find out predictors of selfcare practice. Thematic inductive analysis was done to explore constraints. Results: 73% study participants were aware about overall selfcare practices; 43.8% performed satisfactory overall selfcare practice. Below secondary level educational status [AOR:0.412, 95% CI (0.178-0.956)] and lower & lower-middle socioeconomic status [AOR:0.063, 95% CI (0.017-0.230)] were found to create negative influence on favourable self-care practice. Thematic analysis revealed false belief, lack of motivation, lack of proper knowledge, emotional disturbances and Covid-19 pandemic impact as major constraints in selfcare practice. Conclusion: Selfcare awareness & practice both are low. Interventions are needed to ensure healthy lifestyle of patients.
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Background: Rhabdomyosarcoma (RMS) is the most common soft tissue sarcoma in children. Anaplasia is a rare phenomenon seen in childhood RMS. The most common histologic subtype was Embryonal followed by Alveolar and spindle subtype. Design: A total of 11 cases of pediatric RMS were selected from January 2017 to June 2019 presenting at various sites. Out of 11 cases, 2 were further diagnosed as Embryonal, 2 as Alveolar, 2 as Pleomorphic, 1 as Spindle subtype and rest 4 as RMS-NOS based on morphology. All cases were positive for Desmin. The presence of cells with lobated, hyperchromatic nuclei at least three times larger than the tumor cell (anaplastic cells) was selected as the main criterion to diagnose Anaplasia. Results: Out of the total 11 cases, anaplasia was seen in 7 cases. Out of these seven, five cases showed Focal Anaplasia (FA) (71.4%) and 2 cases showed Diffuse Anaplasia (DA) (28.6%). Out of 2 cases of Embryonal RMS one exhibited focal anaplasia (50%). One case of Spindle RMS showed diffuse anaplasia, 2 cases of pleomorphic RMS showed focal anaplasia. Out of 3 cases of RMS- NOS, 2 exhibited focal anaplaisa and one displayed Diffuse anaplasia. Both Alveolar RMS had no features of anaplasia. Conclusion: Presence of Anaplasia is a frequent observation in pediatric RMS. Anaplasia is often under reported in pediatric RMS. Pathologist should be more aware of this rare phenomenon.
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Background: Resistance to antibiotics is rising markedly. Factor which contributes to resistance is the rampant irrational use of antibiotics. The trend of prescribing antibiotics in intensive care units (ICUs) of eastern India is less explored. Aims and Objectives: The aim of the study was to describe and analyze the utilization of antibiotics as per the WHO/INRUD prescribing core indicators in an intensive and critical care unit (CCU) of a tertiary center in eastern India. Materials and Methods: A prospective observational study was carried out on prescription pattern of antibiotics. Case records of patients with restricted antibiotic therapy were reviewed and evaluated using descriptive statistics. A total of 353 prescriptions were evaluated and analyzed. Results: Among the total 353 patients most common age group admitted was 41–60 years. Males were more in numbers. Myocardial infarction and post-operative complications were the most common cause of admission in ICU and CCU, respectively. Ceftriaxone (44%) and meropenem (37%) were the drugs used rampantly in ICU and CCU. More than 90% of prescriptions had injections and drugs were written in generic names. Almost 90% of patients in CCU had antibiotics in their prescription. Adverse events occurred in 14.7% of patients and thrombophlebitis was the most common adverse event occurred. Conclusion: The study has given us an overall impression of the antibiotics usage pattern in ICU and CCU of this teaching institution. The study has shown that antibiotics should be used judiciously in ICU and CCU. Policy can be made on the basis of the result of this study.
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Background: Primary Breast Sarcomas (PBS) are rare malignancies and seen in less than <1 % of all breast malignancies. PBS are non epithelial, composed of mesenchymal mammary tissue and are difficult to diagnose from other sarcomas arising in breast. Materials and Methods: A retrospective study was conducted in the Department of Pathology and slides of breast malignancies over a period of 5 years were reviewed. Out of total 1570 breast malignancies, 5 cases were reported as PBS. Diagnosis was made on the basis of Histopathology and IHC findings. Results: Out of total 1570 cases, 5 cases were diagnosed as PBS (i.e. 0.32% of all cases). 3 out of 5 cases were males comprising of 60% of cases and 2 cases were females accounting for 40% of cases. The age group of presentation was 32-65 years with mean age being 48.5 years. A diagnosis of MPNST was rendered in two cases ( 1=M, 1=F), one each was diagnosed as DFSP ( with fibrosarcoma), Leiomyosarcoma and Fibrosarcoma. Conclusion: PBS is an extremely rare entity and locally aggressive. It requires diagnosis as its treatment protocol is different.
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Introduction: Health workforce is the important pillar of health system in India. Efficient utilization of scarce community‑level human resources in health care is critical to ensure optimum care in a community. Objective: To describe the time utilization pattern of health workers (HWs) during field activity at the primary healthcare level and to study the facilitators and barriers in efficient time utilization. Methods: A mixed‑method study with a time and motion approach was carried out in the rural areas of Ballabgarh block of Haryana. Time and motion approach was continuous, direct, synchronous and both active and passive method of time logging was adapted. Electronic Geo‑Positioning System based mobile, timestamp application and the digital stopwatch were used to capture time utilization. The quality of the data collected in the daily work plan during the household (HH) visits was assessed using semi‑structured interview schedule. In‑depth interview with the HWs was carried out to understand the facilitating factors and barriers in their efficient functioning. Results: Proportion of HH s covered in data collection for time and motion patterns was 36% out of the total number of HH s in the SCs. The completeness of work plan was 74.8%. The average number of HHs covered per day by an HW was 38. Mean (standard deviation [SD]) time duration spent in each HH in completing the work plan was 2.9 (0.8) minutes. Mean (SD) total distance travelled by HW per day was 1845.1 (974.2) metres. Mean (SD) time duration spent idle in the field was 22.7 (13.0) minutes. Proportion of effective time utilised in the field was 54.3%. Several enabling factors and barriers were identified at personal, community, and health system level. Conclusion: Only half of the allotted time was effectively utilized by the HWs for house visit in the community. Planning the beat schedule as per the feasibility and ensuring quality of HH visits using information technology is critical for providing primary healthcare at village level.
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Anabaena sp. are the dominant cyanobacterial species on terracotta monuments of Bishnupur which exposed to high solar radiation, ultraviolet and in a desiccated condition in most part of the year. In the present study three Anabaena species were isolated from crust samples and its antibacterial activities were evaluated against pathogenic bacteria Bacillus subtilis, Listeria monocytogenes, Staphylococcus aureus, Salmonella typhimurium, Escherichia coli. We observed good antibacterial activity in ethyl acetate and ethanol extract of Anabaena sp. (VBCCA 052002) which having highest antibacterial activity against Staphylococcus aureus, Salmonella typhimurium, Staphylococcus aureus and E. coli respectively. We have validated the antibacterial assay by using resazurin based antimicrobial assay in microtiter plates and calculated the MIC value of ethyl acetate extract of Anabaena sp. (VBCCA 052002) which is found to be 100 µg/ml against Staphylococcus aureus and 150 µg/ml against Salmonella typhimurium.
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Aims of the study: Helicobacter pylori (H. pylori) induces inflammatory changes in the gastric mucosa variably correlated to different endoscopic and histologic features. The present prospective study aimed to correlate different endoscopic findings with histomorphological changes and the presence of H. pylori in the gastro-duodenal mucosa, in samples of dyspeptic patients. Methods: 60 dyspeptic patients of 18 years to 60 years of age were selected from outpatient department and screened with gastro-duodenoscopy and biopsy. The presence of H. pylori was determined by urease test on fresh biopsy specimens and histologically using the modified Giemsa stain. Findings were recorded and analyzed statistically. Results: Highest (84.6%) H. pylori positivity was seen in the 41-50 years age group. Majority of the patients had a normal upper gastro-intestinal endoscopy; among them majority (61.2%) was positive for H. pylori infection. Most cases with endoscopic lesion in the gastro-duodenal mucosa were also positive for H. pylori infection. On biopsy, chronic gastritis was the most common (73.33%) finding in 44 cases, among them, more than two-third (70.4%) were positive for H. pylori. Conclusion: H. pylori gastritis is strongly associated with peptic ulcer diseases, chronic gastritis and non-ulcer dyspepsia. Endoscopy and biopsy play the main role in diagnosis and identification of the spectrum of involvement.
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Objective: Nasal obstruction due to deviated nasal septum is a common problem encountered by otolaryngologist. The standard surgical treatment for symptomatic deviated septum is septoplasty which has gone through several modifications since its inception. Study objectives were to compare the endoscopic and conventional septoplasty and to evaluate the advantage, disadvantage and complication of both the procedures. Materials and Methods: Prospective observational study was conducted in department of ENT and Head-Neck-Surgery of a tertiary care teaching hospital. Sixty patients undergoing either endoscopic septoplasty or conventional septoplasty were studied prospectively for a period of 3 months to compare the efficacy of both the techniques. Objective assessment was done by doing nasal endoscopy 90 days after the operation to note the following points- (1) Persistence of deviation (2) Spur (3) Formation of synechiae (4) Septal perforation. Result: In this study the endoscopic approach showed better overall clinical result as compared to conventional technique with lesser complication. It was noted that endoscopic septoplasty group had minimum blood loss and shorter operative time than conventional method, but difference was not statistically significant. Conclusion: Endoscopic septoplasty was founded with distinct advantage over conventional method due to better illumination, improve accessibility to remote area was founded. Further surgical experience and larger similar studies will help in coming to a greater consensus.
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Aims and Objectives: We examined the prognostic value of Tumor stroma ratio (TSR) in breast tumor core biopsy (TCB) specimen to determine response to neoadjuvant therapy (NAT) prior to modified radical mastectomy (MRM). Methods: This was a retrospective analysis of patients with breast cancer who underwent TCB before NAT between August 2016 and July 2018. TSR in TCB was studied independently by 2 pathologists ( VM, VS) defined as stroma rich (TSR?50%) or stroma poor (TSR>50%). MRM specimen of these patients were subsequently studied .Residual cancer burden (RCB) was calculated using the MD Anderson RCB calculator, categorized as complete (0), good (1) Partial (2) and no response (3). Statistical analysis was done to assess correlation of TSR to RCB. Results: A total of 62 patients were analyzed. Mean(SD) age was 48(11) years.Twenty eight (45%) and 34 (55%) patients were stroma rich and stroma poor respectively. Twenty six (42%) patients were responders and 36 (58%) non-responders to NAT. Among stroma rich patients, only 3 (10%) were responders (Class 0 &1)and 25 (90%) non-responders(Class2&3)to NAT, among stroma poor patients 23 (68%) responded well and 11 (32%) did not.TSR had a moderate negative correlation with RCB (-0.6). On univariate analysis, only TSR had a significant effect on RCB class (<0.001). Conclusions: TSR on TCB is a useful prognostic factor to determine response of breast carcinoma patients to neoadjuvant therapy.It is cost effective, simple and quick. Larger multi-centric studies would be useful to study its clinical implications.
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Background: In adults most common intracranial malignant lesion is brain metastasis, far outnumbering primary brain tumor. The most common primary site is lung cancer (18–64%), followed by breast (25–21%), malignant melanoma (4–16%), and colorectal cancer (2–12%). It is hypothesized that the incidence of brain metastasis might be increasing, as a result of increasing survival from recent advance in cancer treatment, more frequent brain screening for specific primary malignancy that known to have a higher prediction for brain metastasis and greater availability and use of magnetic resonance imaging (MRI) of brain. In clinical oncology, understanding brain metastasis is important, because it has profound effect on length of survival, quality of life, and in one-third to one-half of affected patients, they represent the direct cause of death despite current improvement in therapeutic approach. Epidemiological data of brain metastasis are lacking in India. Objectives: Aims of our retrospective analysis are to study epidemiology and pattern of care of brain metastasis over last one decade in Nil Ratan Sircar Medical College and Hospital, Kolkata. Materials and Methods: Between 2006 and December 2017, a total of 710 patients of brain metastasis treated in our department with palliative intent were analyzed retrospectively. New-onset neurological symptoms in a known case of cancer we always presumed that, symptoms were due to brain metastasis until proven otherwise. Hence, all patients presenting with acute neurological signs and symptoms underwent through clinical examination, contrast-enhanced (CE) computed tomography brain, and/or CEMRI of brain. Epidemiology, pattern of care, and outcome in the form of overall survival (OS) and disease-free survival were determined. Results: Fifty-seven percent patients were male. The median age was 62 years at the time of diagnosis. Lung carcinoma was most common primary site seen in 52% patients, followed by carcinoma breast second most common primary site, seen in 32% patients. Headache (73%) and motor weakness were most common presenting symptoms. Supratentorial location most common site, out of which parietal region is most common. The only small number of patients was offered best supportive care alone whereas majority of the patients were considered fit for palliative therapy. Treatment consisted of metastasectomy when possible and palliative whole-brain radiotherapy (WBRT) alone or followed by systemic therapy. Optimal supportive care in addition to chemotherapy or radiotherapy is given to all patients. A total of 254 patients were given blood product, erythropoietin, granulocyte-colony-stimulating factor following chemotherapy. Hospitalization required in 71% patients and tumor-related problem was most common cause (46%). Remaining patients were hospitalized for delivery WBRT or CCT. The median OS is 9 months. Patients with younger age and breast primary associated with better prognosis than lung primary. Conclusions: We can conclude that carcinoma lung in male and carcinoma breast in females was most common cause of brain metastasis. Because advance in palliative therapy, outcome of patients with brain metastasis has improved, and patients with brain metastasis benefit from palliative radiotherapy and chemotherapy and this treatment could be delivered easily on outpatients basis.
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Background: Worldwide, an estimated 572,034 esophageal cancer cases and 508,585 deaths occurred in 2018 and it accounts for approximately 3.2% of all malignancy. Because esophagus has no serosal covering with extensive, longitudinal connecting system of lymphatic plexus, direct invasion to contiguous structures and lymph node metastasis occurs early. Unresectable or metastatic disease at the time of diagnosis is seen in approximately 80% of patients, with cure rate <15% and thus making carcinoma of esophagus is one of the most dreaded malignancies. As most of the patients are diagnosed in locally advanced or metastatic stage, so curative surgical resection is not an option. Hence, in these groups of patients, other treatment modalities including concurrent chemoradiation have been tried. However, many of these patients are in a poor general condition so that radical concurrent chemoradiation as an alternative surgical resection could not be offered. In this group of patients, only radiotherapy (RT) is an option in intention to improve quality of life and to increase disease-free survival (DFS) if possible. As there is more chance local failure when patients treated with only external beam RT (EBRT), increasing dose to tumor may improve local control. Intraluminal brachytherapy (ILBT) is an important treatment option for dose escalation along with EBRT in the treatment of locally advanced and inoperable carcinoma esophagus. ILBT provides focal dose escalation, rapid reduction tumor, rapid restoration of swallowing function with sparing of surrounding normal tissue, and potentially improving therapeutic ratio. Hence, based on these facts, following EBRT, ILBT is an effective adjuvant modality to delivered high tumoricidal dose which can facilitate good local control, DFS with acceptable toxicity. We have used ILBT alone as palliative RT and combined modality with EBRT as radical treatment approach. Objectives: The aim of our study is to evaluate efficacy and safety of external beam radiation plus ILBT in locally advanced, inoperable carcinoma of esophagus in terms of improving local control, DFS, toxicity, and quality of life. Materials and Methods: A total of 58 carcinoma esophagus patients treated with EBRT plus ILBT in our RT department from 2012 to 2015 analyzed retrospectively. EBRT, total dose of 40 Gy/20 fractions, delivered in 4 weeks, using anteroposterior posteroanterior portal in cobalt-60 machine. Two–three weeks after completion of EBRT, ILBT was done using esophageal budgie. The total dose of brachytherapy was 10 Gy in two fractions, 1 week apart, 5 Gy in each fraction. EBRT and ILBT treatment completed in 8–9 weeks. Response assessed by clinical assessment, upper gastrointestinal endoscopy, and contrast-enhanced computed tomography chest and abdomen initially at 3 months and then at 6 months. Results: Local disease control seen in 65% of patients. With a median follow-up of 15 months, the median DFS was 8 months and median overall survival was 14 months. Regional nodal failure and distant metastasis were seen in 35% and 46% of patients, respectively. The incidence of acute mucositis was seen in 75% of patients and late toxicity is seen in 25% of cases. Swallowing function preserved in >87% of patients. Conclusion: In patients with locally advanced carcinoma of esophagus and poor performance status who are unable to tolerate radical concurrent chemoradiation, combination of EBRT plus ILBT produces good local control, DFS, and durable relief of dysphagia with acceptable toxicity.
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Background: Pleural effusion has varied aetiological factors. It constitutes one of the major causes of morbidity in India as well in other parts of world. Because of the various aetiologies that can cause pleural effusion, itoften present a diagnostic problem, even after extensive investigations. Objective: In this study, authors aimed to identify the common aetiologies causing pleural effusion and their clinical profile in a tertiary care hospital. Materials and Methods: A hospital based cross-sectional study is conducted over a period of one year in tertiary care hospital in West Bengal. 150 patients of pleural effusion above 10 yrs of age were studied. Clinico-pathological, radiological, hematological and biochemical parameters were documented. Results: The most common cause pleural effusion in this study was tuberculosis (64.67%), followed by malignancy (14.67%), parapneumonic effusion (7.33%), cardiac failure (5.33%) and other minor causes. It was commonly seen in male (70%). The occurrence of tubercular pleural effusion was maximum in the age group 31-40 years. Right-sided effusions were more common. Pleural fluid cytology and adenosine deaminase played a pivotal role in the diagnosis of tubercular pleural effusion. Conclusion: The present study highlights tuberculosis as the common causative factor for pleural effusion, labels lung carcinoma as the most common cause of malignant pleural effusion, and defines the clinico-pathological, biochemical and imaging characteristics of different aetiologies of pleural effusion.
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Background: Cytological examination of pericardial effusion fluids is important in diagnosing the etiology of underlying disease, staging, and prognosis of cancer. Aims and Objectives: (1) To study cytological evaluation of pericardial effusions in various pathological conditions in a tertiary care center. (2) To analyze their frequency and clincopathological correlation of the diagnosis. Materials and Methods: Our study was a retrospective study performed in the Department of Pathology from 1st January 2012 to 31st December 2016. The study sample included all the pericardial effusions submitted in the pathology department for cytological evaluation. Clinical details and relevant parameters correlated with clinical findings. Each fluid underwent cytospin and cytocentrifuge along with preparation of conventional smears. Results: Of 120 cases, 80% were of benign effusion and 20% were of malignant effusion. Male-to-female ratio was 1.44:1 with patient age ranging from 3 to 90 years. Conclusion: Benign effusions can been seen in younger age group and malignant ones in the older age group. The preliminary pericardial fluid analysis in resource-limited settings is the most convenient and cost-effective method for accurate diagnosis. It reduces the demand of invasive investigations and its complications. At times, it is the first test to point toward underlying malignant process thereby affecting the prognosis, survival, and treatment outcome of the patient.
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Background & objectives: To support recent political commitments to end tuberculosis (TB) in the World Health Organization South-East Asian Region (SEAR), there is a need to understand by what measures, and with what investment, these goals could be reached. These questions were addressed by using mathematical models of TB transmission by doing the analysis on a country-by-country basis in SEAR. Methods: A dynamical model of TB transmission was developed, in consultation with each of the 11 countries in the SEAR. Three intervention scenarios were examined: (i) strengthening basic TB services (including private sector engagement), (ii) accelerating TB case-finding and notification, and (iii) deployment of a prognostic biomarker test by 2025, to guide mass preventive therapy of latent TB infection. Each scenario was built on the preceding ones, in successive combination. Results: Comprehensive improvements in basic TB services by 2020, in combination with accelerated case-finding to increase TB detection by at least two-fold by 2020, could lead to a reduction in TB incidence rates in SEAR by 67.3 per cent [95% credible intervals (CrI) 65.3-69.8] and TB deaths by 80.9 per cent (95% CrI 77.9-84.7) in 2035, relative to 2015. These interventions alone would require an additional investment of at least US$ 25 billion. However, their combined effect is insufficient to reach the end TB targets of 80 per cent by 2030 and 90 per cent by 2035. Model projections show how additionally, deployment of a biomarker test by 2025 could end TB in the region by 2035. Targeting specific risk groups, such as slum dwellers, could mitigate the coverage needed in the general population, to end TB in the Region. Interpretation & conclusions: While the scale-up of currently available strategies may play an important role in averting TB cases and deaths in the Region, there will ultimately be a need for novel, mass preventive measures, to meet the end TB goals. Achieving these impacts will require a substantial escalation in funding for TB control in the Region.
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Trophoblastic differentiation of endometrial carcinoma is extremely rare, till date 18 cases reports are there in the literature. A 68-year-old postmenopausal female presented with abnormal vaginal bleeding. Histopathologically, there were areas of serous carcinoma with trophoblastic differentiation (~90%). On immunohistochemistry, the trophoblastic component was positive for ?-human chorionic gonadotropin (hCG), HPL and EMA. IHC confirmed the diagnosis of serous carcinoma with trophoblastic differentiation. The clinicopathological features of 18 previously reported cases of trophoblastic differentiation in the uterine tumor were analyzed in addition to the present case.
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A quantitative study was undertaken to determine the expression level of different effl ux pumps in multi-drug-resistant (MDR) Escherichia coli isolates from urinary infection. We have determined the presence of different effl ux pumps and measured the expression level of tolC, mdfA, norE and ynfA genes among 48 isolates by quantitative real-time PCR. The expression level of tolC and ynfA was constantly high and observed among 75-80% of isolates, whereas mdfA and norE were expressed occasionally. Our fi ndings suggest that ynfA, a new SMR effl ux pump gene family member increases the antibiotics’ resistance in E. coli.
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Bacterial L-asparaginase has been widely used as therapeutic agent in treatment of various lymphoblastic leukemia and food processing aid to reduce the formation of cancer causing acrylamide. The present work deals with production and purification of extracellular L-asparaginase from soil isolates using solid state fermentation. The isolate was characterized by big chemical test and identified as Bacillus subtilis. The enzyme production was carried out by solid state fermentation comparing the results with submerged fermentation. The enzyme was purified to near homogeneity by ammonium sulphate precipitation, dialysis, followed by gel filtration on Sephadex G-100 column, CM Sephadex C-50 and SDS-PAGE. The enzyme was purified at 110.2 folds and showed a final specific activity of 1785.7 IU/mg with 26.5% yield. SDS-PAGE of the purified enzyme revealed an apparent molecular weight of 109 kDa. The purified enzyme showed maximum activity at pH 9 when it was incubated at 50°C for 35 min. The enzyme was activated by Mg+2 and strongly inhibited by EDTA.
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This paper describes the studies on the purification and partial characterization of serine alkaline protease produced through submerged fermentation process from a locally isolated Bacillus subtilis. This strain, grown in a highly alkaline medium (pH 10), produces an extracellular proteolytic enzyme. The alkaline protease was purified in a simple two-step procedure involving ammonium sulphate precipitation and gel filtration. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) analysis of the purified alkaline protease indicated an estimated molecular mass of 30KDa. It was more active in the range of 20-60ºC and had an optimum activity at 55ºC with optimum pH of 10.5. Characterization of the protease showed that it required certain cations such as Mg++, Mn++ and Ca++ for maximal activity. The serine nature of the alkaline protease was confirmed by PMSF inhibition. The temperature and pH stability of this Alkaline Protease from Bacillus Subtilismakes it potentially useful forindustrial applications.