Differential expression of inflammatory cytokines, prostaglandin synthases and secretory leukocyte protease inhibitor in the endometrium and circulation in different graded CEH-pyometra in bitch.

Cystic endometrial hyperplasia (CEH)-pyometra (CEH-P) is one of the most common reproductive disorders in bitches, posing a risk to both future fertility and life. The aims of the current study were to elucidate the differential expression patterns of inflammatory mediators at transcript and protein levels in the endometrium and to assess the concentrations of key inflammatory mediators in the peripheral circulation of bitches with different graded CEH-P. A total of 25 client-owned intact mixed breed bitches of 3-10 years presented to the outpatient department of RVP-TVCC of the institute were considered for the study. Of which, 22 cases suggestive of pyometra and 3 cases of CEH obtained during routine elective ovariohysterectomy were subjected to histopathological examination. Uteri were categorized into CEH (n = 3), moderate CEH-P (mCEH-P, n = 9), severe CEH-P (sCEH-P, n = 6) and atrophic pyometra (AT-P, n = 7). A group of age matched (n = 12) bitches without pyometra served as control. Endometrial transcripts such as IL6, IL8, PTGS2, PGFS, and SLPI were expressed differentially in the CEH and CEH-P bitch. In addition, a strong immunoreactivity (IR) of IL6, IL8, PTGS2, and mPGES1 was recorded in the sCEH-P uterus, while expression of IL10 was noticed in AT-P. In circulation, serum IL6 was the most relevant marker with high sensitivity of 96.2% and specificity of 84.6% at a cut off concentration 8.5 pg/mL followed by SLPI with 95.2% sensitivity, and 84.6% specificity at cut off concentration of 1.3 ng/mL. Serum IL10, PGFM and SLPI concentration in the peripheral circulation were 1.5-2.23 fold higher in mCEH-P, 0.87-2.5 fold higher in sCEH-P and 2.9-3.5 fold higher in AT-P than that of control. It is concluded that monitoring the serum concentration of IL6, IL10 and SLPI would be useful adjunct to the established hematobiochemical parameters in the management of pyometra in the bitch with critical illness.

Escherichia coli expression and characterization of alfa-amylase from Geobacillus thermodenitrificans DSM-465 / Expressão e caracterização de Escherichia coli de alfa-amilase de Geobacillus thermodenitrificans DSM-465

Alpha amylase, catalyzing the hydrolysis of starch is a ubiquitous enzyme with tremendous industrial applications. A 1698 bp gene coding for 565 amino acid amylase was PCR amplified from Geobacillus thermodenitrificans DSM-465, cloned in pET21a (+) plasmid, expressed in BL21 (DE3) strain of E. coli and characterized. The recombinant enzyme exhibited molecular weight of 63 kDa, optimum pH 8, optimum temperature 70°C, and KM value of 157.7µM. On pilot scale, the purified enzyme efficiently removed up to 95% starch from the cotton fabric indicating its desizing ability at high temperature. 3D model of enzyme built by Raptor-X and validated by Ramachandran plot appeared as a monomer having 31% α-helices, 15% β-sheets, and 52% loops. Docking studies have shown the best binding affinity of enzyme with amylopectin (∆G -10.59). According to our results, Asp 232, Glu274, Arg448, Glu385, Asp34, Asn276, and Arg175 constitute the potential active site of enzyme.

A alfa-amilase, que catalisa a hidrólise do amido, é uma enzima ubíqua com imensas aplicações industriais. Um gene de 1698 pb que codifica a amilase de 565 aminoácidos foi amplificado por PCR, a partir de Geobacillus thermodenitrificans DSM-465, clonado no plasmídeo pET21a (+), expresso na cepa BL21 (DE3) de E. coli e caracterizado. A enzima recombinante exibiu peso molecular de 63 kDa, pH ótimo igual a 8, temperatura ótima de 70° C e valor KM de 157,7 µM. Em escala piloto, a enzima purificada removeu com eficiência até 95% de amido do tecido de algodão, indicando sua capacidade de desengomagem em alta temperatura. O modelo 3D da enzima construída por Raptor-X e validada por Ramachandran plot apareceu como um monômero com 31% de hélices alfa, 15% de folhas beta e 52% de loops. Os estudos de docking mostraram melhor afinidade de ligação da enzima com amilopectina (∆G: - 10,59). De acordo com nossos resultados, Asp 232, Glu274, Arg448, Glu385, Asp34, Asn276 e Arg175 constituem o sítio ativo potencial da enzima.

Studies on the recombinant production and anticancer activity of thermostable L- asparaginase I from Pyrococcus abyssi / Estudos sobre a produção recombinante e atividade anticancerígena da L asparaginase I termoestável de Pyrococcus abyssi

L-Asparaginase catalysing the breakdown of L-Asparagine to L-Aspartate and ammonia is an enzyme of therapeutic importance in the treatment of cancer, especially the lymphomas and leukaemia. The present study describes the recombinant production, properties and anticancer potential of enzyme from a hyperthermophilic archaeon Pyrococcus abyssi. There are two genes coding for asparaginase in the genome of this organism. A 918 bp gene encoding 305 amino acids was PCR amplified and cloned in BL21 (DE3) strain of E. coli using pET28a (+) plasmid. The production of recombinant enzyme was induced under 0.5mM IPTG, purified by selective heat denaturation and ion exchange chromatography. Purified enzyme was analyzed for kinetics, in silico structure and anticancer properties. The recombinant enzyme has shown a molecular weight of 33 kDa, specific activity of 1175 U/mg, KM value 2.05mM, optimum temperature and pH 80°C and 8 respectively. No detectable enzyme activity found when L-Glutamine was used as the substrate. In silico studies have shown that the enzyme exists as a homodimer having Arg11, Ala87, Thr110, His112, Gln142, Leu172, and Lys232 being the putative active site residues. The free energy change calculated by molecular docking studies of enzyme and substrate was found as ∆G – 4.5 kJ/mole indicating the affinity of enzyme with the substrate. IC50 values of 5U/mL to 7.5U/mL were determined for FB, caco2 cells and HepG2 cells. A calculated amount of enzyme (5U/mL) exhibited 78% to 55% growth inhibition of caco2 and HepG2 cells. In conclusion, the recombinant enzyme produced and characterized in the present study offers a good candidate for the treatment of cancer. The procedures adopted in the present study can be prolonged for in vivo studies.

A L-asparaginase, que catalisa a degradação da L-asparagina em L-aspartato e amônia, é uma enzima de importância terapêutica no tratamento do câncer, especialmente dos linfomas e da leucemia. O presente estudo descreve a produção recombinante, propriedades e potencial anticancerígeno da enzima de Pyrococcus abyssi, um archaeon hipertermofílico. Existem dois genes que codificam para a asparaginase no genoma desse organismo. Um gene de 918 bp, que codifica 305 aminoácidos, foi amplificado por PCR e clonado na cepa BL21 (DE3) de E. coli usando o plasmídeo pET28a (+). A produção da enzima recombinante foi induzida sob 0,5mM de IPTG, purificada por desnaturação seletiva por calor e cromatografia de troca iônica. A enzima purificada foi analisada quanto à cinética, estrutura in silico e propriedades anticancerígenas. A enzima recombinante apresentou peso molecular de 33 kDa, atividade específica de 1.175 U / mg, valor de KM 2,05 mM, temperatura ótima de 80º C e pH 8. Nenhuma atividade enzimática detectável foi encontrada quando a L-glutamina foi usada como substrato. Estudos in silico mostraram que a enzima existe como um homodímero, com Arg11, Ala87, Thr110, His112, Gln142, Leu172 e Lys232 sendo os resíduos do local ativo putativo. A mudança de energia livre calculada por estudos de docking molecular da enzima e do substrato foi encontrada como ∆G – 4,5 kJ / mol, indicando a afinidade da enzima com o substrato. Valores de IC50 de 5U / mL a 7,5U / mL foram determinados para células FB, células caco2 e células HepG2. Uma quantidade de enzima (5U / mL) apresentou inibição de crescimento de 78% a 55% das células caco2 e HepG2, respectivamente. Em conclusão, a enzima recombinante produzida e caracterizada no presente estudo é uma boa possibilidade para o tratamento do câncer. Os procedimentos adotados na presente pesquisa podem ser aplicados para estudos in vivo.

Escherichia coli expression and characterization of -amylase from Geobacillus thermodenitrificans DSM-465

Abstract Alpha amylase, catalyzing the hydrolysis of starch is a ubiquitous enzyme with tremendous industrial applications. A 1698 bp gene coding for 565 amino acid amylase was PCR amplified from Geobacillus thermodenitrificans DSM-465, cloned in pET21a (+) plasmid, expressed in BL21 (DE3) strain of E. coli and characterized. The recombinant enzyme exhibited molecular weight of 63 kDa, optimum pH 8, optimum temperature 70°C, and KM value of 157.7µM. On pilot scale, the purified enzyme efficiently removed up to 95% starch from the cotton fabric indicating its desizing ability at high temperature. 3D model of enzyme built by Raptor-X and validated by Ramachandran plot appeared as a monomer having 31% -helices, 15% -sheets, and 52% loops. Docking studies have shown the best binding affinity of enzyme with amylopectin (G -10.59). According to our results, Asp 232, Glu274, Arg448, Glu385, Asp34, Asn276, and Arg175 constitute the potential active site of enzyme.

Resumo A alfa-amilase, que catalisa a hidrólise do amido, é uma enzima ubíqua com imensas aplicações industriais. Um gene de 1698 pb que codifica a amilase de 565 aminoácidos foi amplificado por PCR, a partir de Geobacillus thermodenitrificans DSM-465, clonado no plasmídeo pET21a (+), expresso na cepa BL21 (DE3) de E. coli e caracterizado. A enzima recombinante exibiu peso molecular de 63 kDa, pH ótimo igual a 8, temperatura ótima de 70° C e valor KM de 157,7 µM. Em escala piloto, a enzima purificada removeu com eficiência até 95% de amido do tecido de algodão, indicando sua capacidade de desengomagem em alta temperatura. O modelo 3D da enzima construída por Raptor-X e validada por Ramachandran plot apareceu como um monômero com 31% de hélices alfa, 15% de folhas beta e 52% de loops. Os estudos de docking mostraram melhor afinidade de ligação da enzima com amilopectina (G: - 10,59). De acordo com nossos resultados, Asp 232, Glu274, Arg448, Glu385, Asp34, Asn276 e Arg175 constituem o sítio ativo potencial da enzima.

Studies on the recombinant production and anticancer activity of thermostable L- asparaginase I from Pyrococcus abyssi

Abstract L-Asparaginase catalysing the breakdown of L-Asparagine to L-Aspartate and ammonia is an enzyme of therapeutic importance in the treatment of cancer, especially the lymphomas and leukaemia. The present study describes the recombinant production, properties and anticancer potential of enzyme from a hyperthermophilic archaeon Pyrococcus abyssi. There are two genes coding for asparaginase in the genome of this organism. A 918 bp gene encoding 305 amino acids was PCR amplified and cloned in BL21 (DE3) strain of E. coli using pET28a (+) plasmid. The production of recombinant enzyme was induced under 0.5mM IPTG, purified by selective heat denaturation and ion exchange chromatography. Purified enzyme was analyzed for kinetics, in silico structure and anticancer properties. The recombinant enzyme has shown a molecular weight of 33 kDa, specific activity of 1175 U/mg, KM value 2.05mM, optimum temperature and pH 80°C and 8 respectively. No detectable enzyme activity found when L-Glutamine was used as the substrate. In silico studies have shown that the enzyme exists as a homodimer having Arg11, Ala87, Thr110, His112, Gln142, Leu172, and Lys232 being the putative active site residues. The free energy change calculated by molecular docking studies of enzyme and substrate was found as G 4.5 kJ/mole indicating the affinity of enzyme with the substrate. IC50 values of 5U/mL to 7.5U/mL were determined for FB, caco2 cells and HepG2 cells. A calculated amount of enzyme (5U/mL) exhibited 78% to 55% growth inhibition of caco2 and HepG2 cells. In conclusion, the recombinant enzyme produced and characterized in the present study offers a good candidate for the treatment of cancer. The procedures adopted in the present study can be prolonged for in vivo studies.

Resumo A L-asparaginase, que catalisa a degradação da L-asparagina em L-aspartato e amônia, é uma enzima de importância terapêutica no tratamento do câncer, especialmente dos linfomas e da leucemia. O presente estudo descreve a produção recombinante, propriedades e potencial anticancerígeno da enzima de Pyrococcus abyssi, um archaeon hipertermofílico. Existem dois genes que codificam para a asparaginase no genoma desse organismo. Um gene de 918 bp, que codifica 305 aminoácidos, foi amplificado por PCR e clonado na cepa BL21 (DE3) de E. coli usando o plasmídeo pET28a (+). A produção da enzima recombinante foi induzida sob 0,5mM de IPTG, purificada por desnaturação seletiva por calor e cromatografia de troca iônica. A enzima purificada foi analisada quanto à cinética, estrutura in silico e propriedades anticancerígenas. A enzima recombinante apresentou peso molecular de 33 kDa, atividade específica de 1.175 U / mg, valor de KM 2,05 mM, temperatura ótima de 80º C e pH 8. Nenhuma atividade enzimática detectável foi encontrada quando a L-glutamina foi usada como substrato. Estudos in silico mostraram que a enzima existe como um homodímero, com Arg11, Ala87, Thr110, His112, Gln142, Leu172 e Lys232 sendo os resíduos do local ativo putativo. A mudança de energia livre calculada por estudos de docking molecular da enzima e do substrato foi encontrada como G 4,5 kJ / mol, indicando a afinidade da enzima com o substrato. Valores de IC50 de 5U / mL a 7,5U / mL foram determinados para células FB, células caco2 e células HepG2. Uma quantidade de enzima (5U / mL) apresentou inibição de crescimento de 78% a 55% das células caco2 e HepG2, respectivamente. Em conclusão, a enzima recombinante produzida e caracterizada no presente estudo é uma boa possibilidade para o tratamento do câncer. Os procedimentos adotados na presente pesquisa podem ser aplicados para estudos in vivo.

Escherichia coli expression and characterization of α-amylase from Geobacillus thermodenitrificans DSM-465 / Expressão e caracterização de Escherichia coli de alfa-amilase de Geobacillus thermodenitrificans DSM-465

Alpha amylase, catalyzing the hydrolysis of starch is a ubiquitous enzyme with tremendous industrial applications. A 1698 bp gene coding for 565 amino acid amylase was PCR amplified from Geobacillus thermodenitrificans DSM465, cloned in pET21a (+) plasmid, expressed in BL21 (DE3) strain of E. coli and characterized. The recombinant enzyme exhibited molecular weight of 63 kDa, optimum pH 8, optimum temperature 70°C, and KM value of 157.7µM. On pilot scale, the purified enzyme efficiently removed up to 95% starch from the cotton fabric indicating its desizing ability at high temperature. 3D model of enzyme built by Raptor-X and validated by Ramachandran plot appeared as a monomer having 31% α-helices, 15% ß-sheets, and 52% loops. Docking studies have shown the best binding affinity of enzyme with amylopectin (∆G -10.59). According to our results, Asp 232, Glu274, Arg448, Glu385, Asp34, Asn276, and Arg175 constitute the potential active site of enzyme.

A alfa-amilase, que catalisa a hidrólise do amido, é uma enzima ubíqua com imensas aplicações industriais. Um gene de 1698 pb que codifica a amilase de 565 aminoácidos foi amplificado por PCR, a partir de Geobacillus thermodenitrificans DSM-465, clonado no plasmídeo pET21a (+), expresso na cepa BL21 (DE3) de E. coli e caracterizado. A enzima recombinante exibiu peso molecular de 63 kDa, pH ótimo igual a 8, temperatura ótima de 70° C e valor KM de 157,7 µM. Em escala piloto, a enzima purificada removeu com eficiência até 95% de amido do tecido de algodão, indicando sua capacidade de desengomagem em alta temperatura. O modelo 3D da enzima construída por Raptor-X e validada por Ramachandran plot apareceu como um monômero com 31% de hélices alfa, 15% de folhas beta e 52% de loops. Os estudos de docking mostraram melhor afinidade de ligação da enzima com amilopectina (∆G: - 10,59). De acordo com nossos resultados, Asp 232, Glu274, Arg448, Glu385, Asp34, Asn276 e Arg175 constituem o sítio ativo potencial da enzima.

Studies on the recombinant production and anticancer activity of thermostable L- asparaginase I from Pyrococcus abyssi / Estudos sobre a produção recombinante e atividade anticancerígena da L-asparaginase I termoestável de Pyrococcus abyssi

L-Asparaginase catalysing the breakdown of L-Asparagine to L-Aspartate and ammonia is an enzyme of therapeutic importance in the treatment of cancer, especially the lymphomas and leukaemia. The present study describes the recombinant production, properties and anticancer potential of enzyme from a hyperthermophilic archaeon Pyrococcus abyssi. There are two genes coding for asparaginase in the genome of this organism. A 918 bp gene encoding 305 amino acids was PCR amplified and cloned in BL21 (DE3) strain of E. coli using pET28a (+) plasmid. The production of recombinant enzyme was induced under 0.5mM IPTG, purified by selective heat denaturation and ion exchange chromatography. Purified enzyme was analyzed for kinetics, in silico structure and anticancer properties. The recombinant enzyme has shown a molecular weight of 33 kDa, specific activity of 1175 U/mg, KM value 2.05mM, optimum temperature and pH 80°C and 8 respectively. No detectable enzyme activity found when L-Glutamine was used as the substrate. In silico studies have shown that the enzyme exists as a homodimer having Arg11, Ala87, Thr110, His112, Gln142, Leu172, and Lys232 being the putative active site residues. The free energy change calculated by molecular docking studies of enzyme and substrate was found as ∆G ­ 4.5 kJ/mole indicating the affinity of enzyme with the substrate. IC50 values of 5U/mL to 7.5U/mL were determined for FB, caco2 cells and HepG2 cells. A calculated amount of enzyme (5U/mL) exhibited 78% to 55% growth inhibition of caco2 and HepG2 cells. In conclusion, the recombinant enzyme produced and characterized in the present study offers a good candidate for the treatment of cancer. The procedures adopted in the present study can be prolonged for in vivo studies.

A L-asparaginase, que catalisa a degradação da L-asparagina em L-aspartato e amônia, é uma enzima de importância terapêutica no tratamento do câncer, especialmente dos linfomas e da leucemia. O presente estudo descreve a produção recombinante, propriedades e potencial anticancerígeno da enzima de Pyrococcus abyssi, um archaeon hipertermofílico. Existem dois genes que codificam para a asparaginase no genoma desse organismo. Um gene de 918 bp, que codifica 305 aminoácidos, foi amplificado por PCR e clonado na cepa BL21 (DE3) de E. coli usando o plasmídeo pET28a (+). A produção da enzima recombinante foi induzida sob 0,5mM de IPTG, purificada por desnaturação seletiva por calor e cromatografia de troca iônica. A enzima purificada foi analisada quanto à cinética, estrutura in silico e propriedades anticancerígenas. A enzima recombinante apresentou peso molecular de 33 kDa, atividade específica de 1.175 U / mg, valor de KM 2,05 mM, temperatura ótima de 80º C e pH 8. Nenhuma atividade enzimática detectável foi encontrada quando a L-glutamina foi usada como substrato. Estudos in silico mostraram que a enzima existe como um homodímero, com Arg11, Ala87, Thr110, His112, Gln142, Leu172 e Lys232 sendo os resíduos do local ativo putativo. A mudança de energia livre calculada por estudos de docking molecular da enzima e do substrato foi encontrada como ∆G ­ 4,5 kJ / mol, indicando a afinidade da enzima com o substrato. Valores de IC50 de 5U / mL a 7,5U / mL foram determinados para células FB, células caco2 e células HepG2. Uma quantidade de enzima (5U / mL) apresentou inibição de crescimento de 78% a 55% das células caco2 e HepG2, respectivamente. Em conclusão, a enzima recombinante produzida e caracterizada no presente estudo é uma boa possibilidade para o tratamento do câncer. Os procedimentos adotados na presente pesquisa podem ser aplicados para estudos in vivo.

Studies on the recombinant production and anticancer activity of thermostable L- asparaginase I from Pyrococcus abyssi.

L-Asparaginase catalysing the breakdown of L-Asparagine to L-Aspartate and ammonia is an enzyme of therapeutic importance in the treatment of cancer, especially the lymphomas and leukaemia. The present study describes the recombinant production, properties and anticancer potential of enzyme from a hyperthermophilic archaeon Pyrococcus abyssi. There are two genes coding for asparaginase in the genome of this organism. A 918 bp gene encoding 305 amino acids was PCR amplified and cloned in BL21 (DE3) strain of E. coli using pET28a (+) plasmid. The production of recombinant enzyme was induced under 0.5mM IPTG, purified by selective heat denaturation and ion exchange chromatography. Purified enzyme was analyzed for kinetics, in silico structure and anticancer properties. The recombinant enzyme has shown a molecular weight of 33 kDa, specific activity of 1175 U/mg, KM value 2.05mM, optimum temperature and pH 80°C and 8 respectively. No detectable enzyme activity found when L-Glutamine was used as the substrate. In silico studies have shown that the enzyme exists as a homodimer having Arg11, Ala87, Thr110, His112, Gln142, Leu172, and Lys232 being the putative active site residues. The free energy change calculated by molecular docking studies of enzyme and substrate was found as ∆G - 4.5 kJ/mole indicating the affinity of enzyme with the substrate. IC50 values of 5U/mL to 7.5U/mL were determined for FB, caco2 cells and HepG2 cells. A calculated amount of enzyme (5U/mL) exhibited 78% to 55% growth inhibition of caco2 and HepG2 cells. In conclusion, the recombinant enzyme produced and characterized in the present study offers a good candidate for the treatment of cancer. The procedures adopted in the present study can be prolonged for in vivo studies.

Comparison between Efficacy of Combined Use of Hyaluronidise and Triamcinolone versus Hyaluronidise, Triamcinolone Acetonide and Oral Colchicine in the Treatment of Oral Submucous Fibrosis.

Oral submucous fibrosis (OSF) is a chronic complex potentially pre-malignant condition caused by chewing areca nut and other irritants. It is an insidious process characterized by Juxta-epithelial deposition of fibrous tissue in the oral cavity and pharynx. OSF is very common in Southeast Asia and also now a days increase in Europe and North America. The aim of this study to compare the effectiveness of intralesional injection of triamcinolone and hyalurunidase versus intralesional injection of triamcinolone plus injection hyalurunidase with oral colchicine. The study included 60 patients of clinically diagnosed case of oral submucous fibrosis. Patients were divided into two Groups A and B. Group A patients received combination intralesionsl injection of triamcinolone acetonide 10mg/ml in 1ml with injection hyalurunidase 1500IU in 2ml with injection 2% lidocaine 7ml. 15 days interval in 3 months and Group B received intralesional injection of triamcinolone acetonide 10mg/ml in 1ml with injection hyalurunidase 1500IU in 2ml with injection 2% lidocaine 7ml in each 15 days interval for 3 months with oral colchicine 0.5mg twice daily for 3 months. Diagnosis based on burning sensation of mouth, blanching of mucosa, ulceration in oral cavity and also reduced mouth opening. Follow up assessment was done at intervals 1st follow up on 21st days after starting of treatment then 2nd follow up after 3 months and last 3rd follow up after 6 months. Before starting of treatment all patients were properly explained about the study and took their written consent. Much more improvement occurred in Group B patients, reducing in burning sensation and also increases in opening of mouth. In both groups blanching mucosae were improved. Treatment regimen of Group B is more effective in increasing mouth opening and improves burning sensation of oral cavity. No side effects were seen in both groups' patients.

Cartilage Ossiculoplasty from Stapes to Tympanic Membrane in One-Stage Intact Canal Wall Tympanoplasty for Cholesteatoma.

This prospective study was conducted at Department of ENT, Mymensingh Medical College Hospital, a tertiary care center in Bangladesh January 2017 to December 2018 to report hearing results and post operative complications of cartilage interposition ossiculoplasty in one-stage intact canal wall (ICW) tympanoplasty for cholesteatoma where ossicular chain is eroded or has to be removed either partially or totally. Total 42 patients underwent Intact canal wall (ICW) tympanoplasty for cholesteatoma with at least intact stapes footplate and in conjunction, cartilage ossiculoplasty was done during the same procedure. Patients were followed up regularly at 1 week, 1 month, 3 month, 6 month and at 1 year as usual follow up protocol to note complications and hearing status in 1 year follow up. In intact stapes suprastructure group, in the preoperative period, the mean air conduction thresholds (AC), bone conduction threshold (BC) and air-bone gap (ABG) were 48.3db, 9.5db and 38.8db respectively. Postoperatively, with a mean follow-up of 12 months, AC, BC and ABG were 27.6db, 9.7db and 17.9db respectively. In missing stapes supra structuregroup, in the preoperative period, the mean air conduction thresholds (AC), Bone conduction threshold (BC) and air-bone gap (ABG) were 57.4db 13.5db and 43.9db respectively. Postoperatively, with a mean follow-up of 12 months, AC, BC and ABG, were 33.9db, 14.2db and 19.7db respectively. For management of cholesteatoma cases, cartilage ossiculoplasty can be done effectively in conjunction with of intact canal wall tympanoplasty in a single setting. Complications are a few and easily manageable. Hearing results are at least as good as with other prosthesis and helps in avoiding subsequent surgery, discomfort and cost to the patients.

Hyperglycemic Episodes Are Associated With Postoperative Infections After Cardiac Surgery.

BACKGROUND AND AIMS: To describe the incidence of and risk factors for postoperative infections and the correlation between postoperative hyperglycemia despite tight blood glucose control with infectious and other complications after contemporary cardiac surgery. MATERIAL AND METHODS: The study comprised 1356 consecutive adult patients who underwent cardiac surgery between January 2013 and December 2014 and were followed up for 6 months. Patients surviving the first 2 days were included in the analysis. Preoperative demographic information, medical history, procedural details, and the postoperative course were recorded. The target range for blood glucose levels was 4-7 mmol/L and repeated arterial blood samples were obtained during the intensive care unit stay. The associations of blood glucose levels during the first postoperative day and the occurrence of postoperative infections and other significant complications were analyzed. RESULTS: Of the study cohort, 9.8% developed infectious complications which were classified as major surgical site infections in 2.2%, minor surgical site infections in 1.1%, lung infections in 2.0%, unclear fever or bacteremia in 0.3%, cannula or catheter related in 2.6%, multiple in 1.5%, and other in 0.2%. The incidence of deep sternal wound infection was 2.0%. Repeated hyperglycemia occurred in 39.7% of patients and was associated with increased rates of postoperative infections, 12.1% versus 8.2%, p = 0.019; stroke, 4.9% versus 1.5%, p < 0.001; and mortality, 6.1% versus 2.1%, p < 0.001, when compared to patients with single or no hyperglycemia. CONCLUSION: Every 10th patient develops infectious complications after cardiac surgery. Repeated hyperglycemia is associated with increased rates of infectious complications, stroke, and mortality.

Long-Term Prognosis and Causes of Death After Pleural Infections.

BACKGROUND AND AIMS: The development of pleural infection may imply a worse state of health and prognosis. The objective of this study was to ascertain the long-term survival and causes of death after pleural infections and to compare them to those of matched controls. MATERIAL AND METHODS: Altogether 191 patients treated for pleural infections at a single University Hospital between January 2000 and December 2008 and 1910 age- and gender-matched controls were included. Survival data and the causes of death for non-survivors were obtained from national databases and compared between the groups. RESULTS: The etiology of pleural infection was pulmonary infection in 70%, procedural complication in 9%, trauma in 5%, malignancy in 4%, other in 7%, and unknown in 5% of patients. The course of treatment was surgical in 82%, drainage only in 12%, and conservative in 5% of included patients. The median follow-up time was 11 years. Mortality rates were 8.4% versus 0.8% during the first 90 days, p < 0.001, and 46.6% versus 24.5% overall, p < 0.001, in patients and controls, respectively. Mortality was significantly higher in patients with pulmonary infection, procedural complication, or malignancy as the etiology of pleural infection. In multivariable analysis, advanced age, previous malignancies, institutional care, alcoholism, and malignant etiology for the infection were associated with inferior survival. Deaths caused by malignancies, respiratory diseases, and digestive diseases were significantly more common in patients than in controls. CONCLUSION: Long-term survival in patients with pleural infections is significantly inferior to that of age and gender-matched controls.

Antioxidant potential analysis of P. ostreatus, P. sajor-caju, P. sapidus and P. columbinus.

The present study was conducted to evaluate the antioxidant activity of ethanol, n-hexane, dichloromethane, ethyl acetate and water extracts of four different Pleurotus spps. P. ostreatus, P. sajor-caju, P. sapidus and P. columbinus. The extraction was performed by classical organic solvent extraction (COSE). The extracts of Pleurotus spps. contained appreciable levels of total phenolic contents (TPC) (0.95-19.49 GAE, mg/g) and total flavonoid contents (0.85-3.73 CE, mg/g). All Pleurotus spps. also contained considerable DPPH radical scavenging activity, showing IC50 (19.15-54.50%) and reducing power (0.50-2.94 nm), respectively. The ascorbic acid content was in the range of (2.90-5.97 mg/g) for all Pleurotus spps. All studied Pleurotus spps. showed potential antioxidant activity. The results of four different Pleurotus spps. extracts showed that they can be used as a good food ingredient, and as a medicinal mushroom for digestive ailments, as well as in the pharmaceutical industry and in cosmetics.

Long-term outcomes of a randomized clinical trial of supervised exercise, percutaneous transluminal angioplasty or combined treatment for patients with intermittent claudication due to femoropopliteal disease.

BACKGROUND: The aim was to compare the long-term outcomes of percutaneous transluminal angioplasty (PTA), a supervised exercise programme (SEP) and combined treatment (PTA + SEP) in patients with intermittent claudication owing to femoropopliteal disease. METHODS: Patients recruited to an RCT comparing these treatments were invited for long-term follow-up from 2010 to 2011. Indicators of limb ischaemia were recorded (ankle : brachial pressure index (ABPI) and treadmill walking distances). Duplex ultrasound imaging was also done. Patients completed Short Form 36 and VascuQol quality-of-life (QoL) questionnaires. RESULTS: Of 178 patients initially recruited to the trial, 139 were alive at the time of follow-up (PTA 46, SEP 47, PTA + SEP 46). Assessments were completed for 111 patients. Median time to follow-up was 5·2 (i.q.r. 3·8-7·4) years. Sixty-nine patients (62·2 per cent) were symptomatic; 18 (16·2 per cent) had experienced a major cardiovascular event since their last follow-up visit. Improvement was observed in ABPI in all groups. QoL outcomes were inconsistent across individual groups. PTA and PTA + SEP groups had a significantly higher ABPI than the SEP group. No significant difference was observed in treadmill walking distances, QoL outcomes, restenosis rates, and new ipsilateral and contralateral lesions on duplex imaging. Patients in all groups required reinterventions (PTA 14, SEP 10, PTA + SEP 6). The total number of reinterventions was higher after PTA (29) compared with SEP (17) and PTA + SEP (9), but failed to reach statistical significance. CONCLUSION: PTA, SEP and combined treatment were equally effective long-term treatment options for patients with claudication owing to femoropopliteal disease. The addition of a SEP to PTA can reduce the rate of symptomatic restenosis and reintervention. Registration number: NCT00798850 (http://www.clinicaltrials.gov).

Reasons for failure to quit: a cross-sectional survey of tobacco use in major cities in Pakistan.

BACKGROUND: Tobacco dependence has been defined as a chronic relapsing disease. Around 5 million annual tobacco-related deaths have been reported worldwide. The majority of smokers want to quit but are not successful. OBJECTIVES: To screen our population for tobacco use, gauge the baseline demographics of tobacco users and assess factors associated with failed attempts to quit. METHODS: Free health camps supervised by a physician were held across two major cities of Pakistan. All consenting participants were administered a questionnaire and had their exhaled carbon monoxide (CO) levels measured. RESULTS: Of 12 969 participants successfully enrolled, the mean age was 31.4 ± 10.0 years. More than three quarters were aged 20-40 years (n = 10 168, 78.4%). The overall average CO level was 12.0 ± 8.0 ppm. The majority of the participants wanted to quit, and nearly everyone had received advice about quitting. The majority had tried smoking cessation pharmacotherapy. Friends/peer pressure (n = 1554, 12%), anxiety (n = 681, 5.3%), tobacco dependence (n = 1965, 15.2%) and stress/mood changes (n = 390, 3.0%) were the most widely observed reasons for failure to quit in study participants. CONCLUSION: The information provided by this study can guide the development of more targeted intervention programmes for smokers who wish to quit.

Study of Commonest Variety of Sinonasal Malignancy and Its Sex Wise Distribution.

This study was done to find the commonest variety of sinonasal malignancy and its association with sex.This cross-sectional study was conducted in the Department of Otolaryngology - Head & Neck Surgery Department, Dhaka Medical College & Hospital and in the Department of Otolaryngology - Head & Neck Surgery Department, Bangabandhu Sheikh Mujib Medical University between January 2009 and December 2009. A total of 146 cases of sinonasal malignancy were consecutively included in the study. The diagnosis was confirmed by histopathology. The mean age was 47.8 years (range: 22-75 years). Over three-quarters (77%) of the patients were male with male to female ratio being 3:1. Nearly one-third (30.8%) of the patients was farmer and over one-third (36.3%) was illiterate. The right sinonasal region was involved in 48.6% cases, left sinonasal region in 39% and both sinonasal region in 12.4% cases. Histopathological diagnosis of sinonasal malignancy revealed that squamous cell carcinoma accounted for 82.9% of sinonasal malignancies, 9.6% adenocarcinoma and the rest were olfactory neuroblastoma, adenoid cystic carcinoma and mucoepidermoid carcinoma. About 96.6% of the tumours involved nasal cavity, 97.9% maxillary sinuses, 17.8% frontal sinuses, 48.6% ethmoidal sinuses and 13% sphenoid sinuses. Over 80% of patients were smoker. Exposure to wood dust was found in 26% of cases. Lather tanning in 4.8% of cases and welding in 4.1% of cases. T staging shows that nearly half (48.6%) of the patients lesion was T2, 29.5% T3, 19.9% T4 and 2.1% T1. None of the patients exhibited lymph node involvement or distant metastasis. Males tend to develop squamous cell carcinoma significantly more than the females with risk of acquiring squamous cell carcinoma being nearly 3(1.1-7.1) times higher in male than that in female (p=0.022). The study concludes that the most common histopathological variety of sinonasal malignancy is squamous cell carcinoma and males are prone to develop this malignancy more frequently than the females.

Etiology of Laryngeal Squamous Cell Carcinoma: Study of 50 Cases in Mymensingh Medical College Hospital.

This cross sectional observational study was done in the Department of ENT & Head-Neck Surgery, Mymensingh Medical College, Bangladesh from January 2013 to July 2014. Fifty (50) cases of carcinoma larynx were purposively selected. Clinically diagnosed cases of carcinoma larynx and histologically proven squamous cell carcinoma were included. Among 50 cases age ranged from 35-75 years with an average age of 58.1 years. Maximum patients were in 5th and 6th decades with male-female ratio 16:1. Most of the patient (78%) came from rural areas and came from low socio-economic condition (58%); maximum patients were cultivator (42%) & illiterate (50%). Smoking was the commonest (64%) personal habit. The other common personal habits were chewing of Betel nut & leaf (44%) and chewing of Tobacco (36%). Most of them have more than one habit.

Recurrent Laryngeal Nerve Injury in Thyroid Surgery: One Year Prospective Study in a Tertiary Care Hospital.

This prospective, cross sectional study was carried out in the Department of Otolaryngology & Head-Neck Surgery, Mymensingh Medical College Hospital, Bangladesh from August 2010 to July 2011. One hundred & twenty eight (128) patients with thyroid swelling who underwent surgery were purposely included. The study was designed to determine the incidence of recurrent laryngeal nerve injury during thyroid surgery. Among the 128 patients, female were predominant 78.90%, majority of the patient were in third decade, 96.87% patients were biochemically euthyroid. Total 5.46% patient had recurrent laryngeal nerve injury. Among them 14.28% had transient and 85.72% had permanent injury. Injury was more in total thyroidectomy done for extensive malignancy.

Outcome of Titanium Elastic Intramedullary Nail in the Treatment of Shaft of Femur Fracture in Children.

Background Femoral-shaft fractures are among the most common fractures of the lower extremity in children. There are several different options for treating femoral-shaft fractures in children. Elastic stable intramedullary nailing (ESIN) has become the standard treatment for fractures of shaft of femur in children for reasons including mini-invasive surgery, no need for casting, early mobilization and discharge as well as growing concerns toward cost-effectiveness. Objective To demonstrate the effectiveness of intramedullary fixation of fracture shaft of femur in skeletally immature children using the titanium elastic intramedullary nails. Method Forty children who underwent fixation with titanium intramedullary nails because of fracture of shaft of femur (Winquist and Hansen type 1 and 2) were reviewed. There were 60% male and 40% female patients and mean follow-up was six months. Time of union, deformity at fracture site, limb length discrepancy, knee range of motion and complications were assessed. Result Average age of the patients was 5.17 years (range 3 to 10). All patients achieved complete healing at a mean 12.8 weeks (range 10 to 20 weeks). Average limb length discrepancy was -0.16 cm (range -1.0 to 1.1 cm) average knee range of motion was 137.55 degrees (range 118 to 152 degrees). Complications were recorded in 13 (31.7%) patients and included: Five malunion which did not show any deformity or functional impairment and eight superficial wound infections which were healed after removal of nail. All patients were active as their pre injury levels at six months follow up. Conclusion Elastic stable intramedullary nailing is the method of choice for the simple pediatrics fracture shaft of femur, as it is minimally invasive and shows good functional and cosmetic results. It allows short hospital stay and quick recovery from pain and is cast-free.