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1.
Braz. j. biol ; 84: e256923, 2024. tab, ilus
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1360219

RESUMO

Naturally occurring mutations in morphogenetic protein 15 (BMP15) are associated with decreased ovulation rate (OR), litter size (LS), and sterility. It is of a great interest to elucidate BMP15 gene in Cholistani sheep breed to uplift socio-economic status and the knowledge of Cholistani sheep breeding in Southern Punjab, Pakistan. In our study, a total of 50 infertile Cholistani sheep aged between 2-6 years and having no blood relation were screened for BMP15 mutations. For this purpose, a high-quality DNA was extracted from the blood of sheep followed by primer designing, Polymerase Chain Reaction (PCR) amplification, DNA sequencing, and in silico analyses. Out of total 50 samples, 9 samples including case 1 (T3), case 2 (T8), case 3 (T17), case 4 (T22), case 5 (T25), case 6 (T33), case 7 (T40), case 8 (T44), and case 9 (T47) were found positive for a variety of already reported and novel BMP15 mutations. Further in silico analyses of the observed mutations have shown the functional impact of these mutations on different characteristics (molecular weight, theoretical PI, estimated half-life, instability index, sub-cellular localization, and 3D confirmation) of the encoded proteins, possibly altering the normal functionality. In a nutshell, findings of this study have confirmed the possible essential role of the BMP15 mutations in the infertility of the Cholistani sheep.


Mutações de ocorrência natural na proteína morfogenética 15 (BMP15) estão associadas à diminuição da taxa de ovulação (TO), tamanho da ninhada (TN) e esterilidade. Estudar a BMP15 na raça Cholistani para elevar o status socioeconômico e o conhecimento da criação de ovinos Cholistani no sul de Punjab, Paquistão. Em nosso estudo, 50 ovelhas Cholistani inférteis sem parentesco sanguíneo foram rastreadas para mutações BMP15. Para tanto, um DNA de alta qualidade foi extraído do sangue dessas ovelhas, seguido de concepção do primer, amplificação da reação em cadeia da polimerase (PCR), sequenciamento de DNA e análises in silico. Do total de 50 amostras, 9, incluindo caso 1 (T3), caso 2 (T8), caso 3 (T17), caso 4 (T22), caso 5 (T25), caso 6 (T33), caso 7 (T40), caso 8 (T44) e caso 9 (T47), foram consideradas positivas para uma variedade de mutações BMP15 novas e já relatadas. Mais análises in silico das mutações observadas mostraram o impacto funcional dessas mutações em diferentes características (peso molecular, PI teórico, meia-vida estimada, índice de instabilidade, localização subcelular e confirmação 3D) das proteínas codificadas, possivelmente alterando a funcionalidade normal. Nossos achados confirmaram o possível papel essencial das mutações BMP15 na infertilidade de ovelhas Cholistani.


Assuntos
Animais , Ovinos , Infertilidade , Mutação/genética
2.
Braz. j. biol ; 83: e248281, 2023. tab
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1350304

RESUMO

Abstract The COVID-19 is a contagious viral disease, was first emerged in Wuhan, China in December 2019 and became the whole world on alert. The mortality rate in top most countries in Asia with special reference to Pakistan has been focused. Since February 26 to September 2020 the total confirmed cases and mortality rate was measured through Wikipedia and the notable journals. Iran is the only country having highest number of deaths (5.73%) followed by Indonesia (3.77%) while Saudi Arabia shows the lowest number of deaths as 1.39%. In Pakistan the first case was confirmed in 26th February, 2020. The nCov-19 has closely related to severe acute respiratory syndrome (SARS) hence SARS COV-2 was named. This virus is responsible for more than 33.9 million deaths in over all the world as of 20th September, 2020. The number of new cases is increasing time to time. Sindh province of Pakistan has reported the highest number of cases till September, 20, 2020 as compared to other parts of the country and has the highest number of death followed by Khyber Pakhtunkhwa. Because of the person to person contact the disease is spreading rapidly. The individuals who has already infected with other diseases like cancer or diabetic etc. are vulnerable. The nCOV-19 is the most contagious due to its mode of transmission. There is still no vaccine is available for the treatment of disease caused by nCoV-2019. It is therefore the only option to control this pandemic is to adopt effective preventive measures.


Resumo A covid-19 é uma doença viral contagiosa, que surgiu pela primeira vez em Wuhan, China, em dezembro de 2019, e deixou o mundo todo em alerta. A taxa de mortalidade na maioria dos principais países da Ásia, com referência especial ao Paquistão, foi enfocada. De 26 de fevereiro a setembro de 2020, o total de casos confirmados e a taxa de mortalidade foram medidos por meio da Wikipedia e de periódicos notáveis. O Irã é o único país com maior número de mortes (5,73%), seguido pela Indonésia (3,77%), enquanto a Arábia Saudita mostra o menor número de mortes, 1,39%. No Paquistão, o primeiro caso foi confirmado em 26 de fevereiro de 2020. O nCov-19 está intimamente relacionado à síndrome respiratória aguda grave (SARS), daí o nome SARS COV-2. Esse vírus é responsável por mais de 33,9 milhões de mortes em todo o mundo em 20 de setembro de 2020. O número de novos casos está aumentando de tempos em tempos. A província de Sindh, no Paquistão, registrou o maior número de casos até 20 de setembro de 2020, em comparação com outras partes do país, e tem o maior número de mortes, seguida por Khyber Pakhtunkhwa. Por causa do contato pessoa a pessoa, a doença está se espalhando rapidamente. Indivíduos que já foram diagnosticados com outras doenças, como câncer ou diabetes, etc. são mais vulneráveis. O nCOV-19 é o mais contagioso devido ao seu modo de transmissão. Ainda não há vacina disponível para o tratamento da doença causada pelo nCoV-2019. Portanto, a única opção para controlar essa pandemia é a adoção de medidas preventivas eficazes.


Assuntos
Humanos , Pandemias , COVID-19 , Paquistão/epidemiologia , China , SARS-CoV-2
3.
Braz. j. biol ; 83: e250179, 2023. graf
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1339372

RESUMO

Abstract Diabetes mellitus (DM) is a non-communicable disease throughout the world in which there is persistently high blood glucose level from the normal range. The diabetes and insulin resistance are mainly responsible for the morbidities and mortalities of humans in the world. This disease is mainly regulated by various enzymes and hormones among which Glycogen synthase kinase-3 (GSK-3) is a principle enzyme and insulin is the key hormone regulating it. The GSK-3, that is the key enzyme is normally showing its actions by various mechanisms that include its phosphorylation, formation of protein complexes, and other cellular distribution and thus it control and directly affects cellular morphology, its growth, mobility and apoptosis of the cell. Disturbances in the action of GSK-3 enzyme may leads to various disease conditions that include insulin resistance leading to diabetes, neurological disease like Alzheimer's disease and cancer. Fluoroquinolones are the most common class of drugs that shows dysglycemic effects via interacting with GSK-3 enzyme. Therefore, it is the need of the day to properly understand functions and mechanisms of GSK-3, especially its role in glucose homeostasis via effects on glycogen synthase.


Resumo O diabetes mellitus (DM) é uma doença não transmissível em todo o mundo, na qual existe nível glicêmico persistentemente alto em relação à normalidade. O diabetes e a resistência à insulina são os principais responsáveis ​​pelas morbidades e mortalidades de humanos no mundo. Essa doença é regulada principalmente por várias enzimas e hormônios, entre os quais a glicogênio sintase quinase-3 (GSK-3) é uma enzima principal e a insulina é o principal hormônio que a regula. A GSK-3, que é a enzima-chave, normalmente mostra suas ações por vários mecanismos que incluem sua fosforilação, formação de complexos de proteínas e outras distribuições celulares e, portanto, controla e afeta diretamente a morfologia celular, seu crescimento, mobilidade e apoptose do célula. Perturbações na ação da enzima GSK-3 podem levar a várias condições de doença que incluem resistência à insulina que leva ao diabetes, doenças neurológicas como a doença de Alzheimer e câncer. As fluoroquinolonas são a classe mais comum de drogas que apresentam efeitos disglicêmicos por meio da interação com a enzima GSK-3. Portanto, é necessário hoje em dia compreender adequadamente as funções e mecanismos da GSK-3, principalmente seu papel na homeostase da glicose via efeitos na glicogênio sintase.


Assuntos
Humanos , Resistência à Insulina , Diabetes Mellitus , Quinase 3 da Glicogênio Sintase , Glucose , Homeostase
4.
Iran J Vet Res ; 23(2): 120-127, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36118607

RESUMO

Background: Listeriosis is a zoonotic disease of humans, animals, birds, fish, and crustaceans worldwide. Domestic animals, especially ruminants, are more susceptible to listeriosis. This infectious disease is caused by Listeria monocytogenes, an intracellular bacterium that can cross blood-brain, placental and intestinal barriers. In Pakistan, the incidence and reliable diagnostic tools for the L. monocytogenes are unidentified in Nili-Ravi buffaloes. Aims: This study was designed to inspect listeriosis in buffaloes through molecular techniques and haemato-biochemical analyses. Methods: A total of 230 samples (115 milk and 115 faecal samples) were collected from symptomatic listeriosis cases in Nili-Ravi buffaloes of 3 geographical districts (Rawalpindi, Faisalabad, and Muzaffargarh) Punjab, Pakistan. These samples were processed for DNA extraction using commercialized kits, and L. monocytogenes was confirmed by conventional PCR. Results: The results revealed that 6.08% and 4.34% of the isolates from milk and faecal samples were found positive for L. monocytogenes, respectively. The phylogenetic analysis of these isolates showed 97-100% similarity to isolates from the USA, Switzerland, Japan, and India. The accession numbers on NCBI GenBank appeared as HF558398 (Switzerland), KP965732 (India), EU372032 (USA), and LC259850 (Japan). Haemato-biochemical examinations showed that the values of WBCs, plasma fibrinogen, ALT, and AST significantly increased (P<0.05) in diseased buffaloes compared to healthy ones. Conclusion: The occurrence of listeriosis in buffaloes urges continuous monitoring and surveillance to prevent this emerging disease in Pakistan.

5.
Ann R Coll Surg Engl ; 2022 Aug 11.
Artigo em Inglês | MEDLINE | ID: mdl-35950970

RESUMO

INTRODUCTION: This systematic review with meta-analysis aimed to compare the robotic complete mesocolon excision (RCME) to laparoscopic colectomy (LC) with (LCME) or without CME (LC non-CME) in postoperative outcomes, harvested lymph nodes and disease-free survival. METHODS: We performed a systematic review with meta-analysis according to PRISMA 2020 and AMSTAR 2 guidelines. RESULTS: The literature search yielded seven comparative studies including 677 patients: 269 patients in the RCME group and 408 in the LC group. The pooled analysis concluded to a lower conversion rate in the RCME group (OR=0.17; 95% CI [0.04, 0.74], p=0.02). There was no difference between the two groups in terms of morbidity (OR=1.03; 95% CI [0.70, 1.53], p=0.87), anastomosis leakage (OR=0.83; 95% CI [0.18, 3.72], p=0.81), bleeding (OR=1.90; 95% CI [0.64, 5.58], p=0.25), wound infection (OR=1.37; 95% CI [0.51, 3.68], p=0.53), operative time (mean difference (MD)=36.32; 95% CI [-24.30, 96.93], p=0.24), hospital stay (MD=-0.94; 95% CI [-2.03, 0.15], p=0.09) and disease-free survival (OR=1.29; 95% CI [0.71, 2.35], p=0.41). In the subgroup analysis, the operative time was significantly shorter in the LCME group than RCME group (MD=50.93; 95% CI [40.05, 61.81], p<0.01) and we noticed a greater number of harvested lymph nodes in the RCME group compared with LC non-CME group (MD=8.96; 95% CI [5.98, 11.93], p<0.01). CONCLUSIONS: The robotic approach for CME ensures a lower conversion rate than the LC. RCME had a longer operative time than the LCME subgroup and a higher number of harvested lymph nodes than the LC non-CME group.

7.
Tech Coloproctol ; 26(10): 821-830, 2022 10.
Artigo em Inglês | MEDLINE | ID: mdl-35804251

RESUMO

BACKGROUND: Literature concerning surgical management of transverse colon cancer is scarce, since many key trials excluded transverse colon cancer. The aim of this study was to evaluate clinical and oncological outcomes comparing open, laparoscopic and robotic transverse colon cancer resection. METHODS: Consecutive patients who underwent elective surgery for transverse colon cancer between December 2005 and July 2021 were included. Data were kept in a prospective database approved by the institutional ethics committee. Primary outcome was overall and disease-free survival. Secondary outcomes included complications, operative time, length of stay and lymph node harvest. Statistical analysis was corrected for age and tumour localisation. RESULTS: Two hundred and forty-six (38 robotic, 71 open and 137 laparoscopic resections) were recruited in this study. There were five conversions during laparoscopic procedures. Operative time was significantly shorter in robotic vs laparoscopic procedures (195 vs 238 min, p = 0.005) and length of stay was shorter in robotic vs laparoscopic and open group (7 vs 9 vs 15 days, p < 0.001). There was no difference in overall complications. R0 resections were similar. Lymph node harvest was highest in the robotic group vs. laparoscopic or open (32 vs. 29 vs. 21, p < 0.001). Overall survival was 97%, 85% and 60% (p < 0.001) and disease-free survival was 91%, 78% and 56% (p < 0.001) for the robotic, laparoscopic and open groups, respectively. CONCLUSIONS: Minimally invasive surgery for transverse colon cancer is safe and offers good clinical and oncological outcomes. Robotic resection is associated with significantly shorter operating times, higher lymph node harvest, lower conversion rate and does not increase morbidity. Differences in disease-free and overall survival should be further explored in randomised controlled trials.


Assuntos
Colectomia , Colo Transverso , Neoplasias do Colo , Laparoscopia , Procedimentos Cirúrgicos Robóticos , Colectomia/efeitos adversos , Colectomia/métodos , Colo Transverso/cirurgia , Neoplasias do Colo/cirurgia , Humanos , Laparoscopia/efeitos adversos , Tempo de Internação , Complicações Pós-Operatórias/epidemiologia , Estudos Retrospectivos , Procedimentos Cirúrgicos Robóticos/efeitos adversos , Resultado do Tratamento
8.
Braz J Biol ; 84: e256923, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35137843

RESUMO

Naturally occurring mutations in morphogenetic protein 15 (BMP15) are associated with decreased ovulation rate (OR), litter size (LS), and sterility. It is of a great interest to elucidate BMP15 gene in Cholistani sheep breed to uplift socio-economic status and the knowledge of Cholistani sheep breeding in Southern Punjab, Pakistan. In our study, a total of 50 infertile Cholistani sheep aged between 2-6 years and having no blood relation were screened for BMP15 mutations. For this purpose, a high-quality DNA was extracted from the blood of sheep followed by primer designing, Polymerase Chain Reaction (PCR) amplification, DNA sequencing, and in silico analyses. Out of total 50 samples, 9 samples including case 1 (T3), case 2 (T8), case 3 (T17), case 4 (T22), case 5 (T25), case 6 (T33), case 7 (T40), case 8 (T44), and case 9 (T47) were found positive for a variety of already reported and novel BMP15 mutations. Further in silico analyses of the observed mutations have shown the functional impact of these mutations on different characteristics (molecular weight, theoretical PI, estimated half-life, instability index, sub-cellular localization, and 3D confirmation) of the encoded proteins, possibly altering the normal functionality. In a nutshell, findings of this study have confirmed the possible essential role of the BMP15 mutations in the infertility of the Cholistani sheep.


Assuntos
Proteína Morfogenética Óssea 15 , Infertilidade , Ovinos , Animais , Proteína Morfogenética Óssea 15/genética , Feminino , Infertilidade/veterinária , Mutação , Ovulação , Paquistão , Ovinos/genética
9.
Ann R Coll Surg Engl ; 104(3): e74-e78, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-34730433

RESUMO

We describe a case report of a robotic total pelvic exenteration (TPE) performed for a locally advanced rectal cancer at our institution in August 2018. Technical details and comparison with published literature are discussed. A 62-year-old patient with a locally advanced low rectal cancer T4N1cM0 with extramural vascular invasion (EMVI) underwent an elective robotic pelvic clearance performed by the urology and colorectal teams. He received neoadjuvant long-course chemo-radiotherapy to downstage the rectal cancer. The primary tumour was T4N1c with involvement of the bladder (trigone area) and prostate. After neoadjuvant therapy, MRI scan showed tumour regression grade 4 (TRG4). The patient underwent single docking totally robotic pelvic clearance. Patient's body mass index (BMI) was 32. The operative time was 400 minutes with the docking time of 15 minutes. There were no intraoperative complications, and the blood loss was 100ml. Histology was ypT4b, ypN1b, ypMx with 2/9 positive lymph nodes, and there was a complete resection by >1mm at all margins. The postoperative complications were ileus and urinary tract infection. Length of stay was 11 days complicated by prolonged ileus requiring total parenteral nutrition (TPN). The 30-day follow-up had no postoperative complications or readmission. The robotic approach is safe and feasible for multiorgan resections for locally advanced pelvic cancers, with curative intent. The literature supports it by highlighting the advantages of robotic pelvic surgery: better access, stable platform, quick inter-specialty change of operator by use of dual console and superior visualisation.


Assuntos
Exenteração Pélvica , Neoplasias Retais/cirurgia , Procedimentos Cirúrgicos Robóticos , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias Retais/diagnóstico por imagem
10.
Environ Pollut ; 294: 118631, 2022 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-34871646

RESUMO

Ultrafine particles (UFP), harmful to human health, are emitted at high levels from motorized traffic. Bicycle commuting is increasingly encouraged to reduce traffic emissions and increase physical activity, but higher breathing rates increase inhaled UFP concentrations while in traffic. We assessed exposure to UFP while cycling along a fixed 8.5 km inner-city route in Copenhagen, on weekdays over six weeks (from September to October 2020), during morning and afternoon rush-hour, as well as morning non-rush-hour, traffic time periods starting from 07:45, 15:45, and 09:45 h, respectively. Continuous measurements were made (each second) of particle number concentration (PNC) and location. PNC levels were summarized and compared across time periods. We used generalized additive models to adjust for meteorological factors, weekdays and trends. A total of 61 laps were completed, during 28 days (∼20 per time period). Overall mean PNC was 18,149 pt/cm3 (range 256-999,560 pt/cm3) with no significant difference between morning rush-hour (18003 pt/cm3), afternoon rush-hour (17560 pt/cm3) and late morning commute (17560 pt/cm3) [p = 0.85]. There was substantial spatial variation of UFP exposure along the route with highest PNC levels measured at traffic intersections (∼38,000-42000 pt/cm3), multiple lane roads (∼38,000-40000 pt/cm3) and construction sites (∼44,000-51000 pt/cm3), while lowest levels were measured at smaller streets, areas with open built environment (∼12,000 pt/cm3), as well as at a bus-only zone (∼15,000 pt/cm3). UFP exposure in inner-city Copenhagen did not differ substantially when bicycling in either rush-hour or non-rush-hour, or morning or afternoon, traffic time periods. UFP exposure varied substantially spatially, with highest concentrations around intersections, multiple lane roads, and construction sites. This suggests that exposure to UFP is not necessarily reduced by avoiding rush-hours, but by avoiding sources of pollution along the bicycling route.


Assuntos
Poluentes Atmosféricos , Material Particulado , Poluentes Atmosféricos/análise , Ciclismo , Dinamarca , Exposição Ambiental/análise , Monitoramento Ambiental , Humanos , Tamanho da Partícula , Material Particulado/análise , Meios de Transporte , Emissões de Veículos/análise
11.
Eur Ann Otorhinolaryngol Head Neck Dis ; 139(2): 83-90, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33685826

RESUMO

There is a controversy in regards to the efficacy of photobiomodulation (PBM) in the management of tinnitus. The aim was to systematically review randomized controlled trials (RCTs) that assessed the efficacy of PBM (low-level laser therapy) in the management of tinnitus. The focused question was "Is PBM effective in the management of tinnitus?". Indexed databases were searched up to and including June 2020 using different combinations of the following key words: (a) laser; (b) diode; (c) low-level laser therapy; (d) photobiomodulation; (e) tinnitus; (f) medium-level laser; (g) photo-biomodulation; and (h) low-power laser; and RCTs performed on humans were included. Letters to the editor; case reports/series; commentaries; experimental studies and historic reviews were excluded. The risk of bias was assessed using the modified cochrane collaboration tool. The format of the current systematic review was personalized to summarize the appropriate information. Ten RCTs (2 single-blinded and 8 double-blinded) were included. One study reported 30% and 100% resolution of tinnitus using diode and Neodymium-doped Yttrium Aluminum Garnet lasers; respectively. One study reported that PBM was effective in relieving tinnitus for up to 3 months. Eight studies reported that PBM was ineffective in the management of chronic tinnitus. The risk of bias was high; medium and low in 4; 5 and 1 studies; respectively. The effectiveness of PBM in the management of tinnitus remains debatable. Further power-adjusted and well-designed RCTs with long-term follow-up are needed.


Assuntos
Terapia com Luz de Baixa Intensidade , Zumbido , Humanos , Zumbido/radioterapia
12.
Braz. j. biol ; 82: e244735, 2022. tab, graf
Artigo em Inglês | LILACS | ID: biblio-1249280

RESUMO

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.


Assuntos
Humanos , Asparaginase/biossíntese , Asparaginase/farmacologia , Pyrococcus abyssi/enzimologia , Antineoplásicos/farmacologia , Especificidade por Substrato , Estabilidade Enzimática , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/farmacologia , Células CACO-2 , Escherichia coli/genética , Simulação de Acoplamento Molecular , Concentração de Íons de Hidrogênio
13.
Braz. j. biol ; 82: e239449, 2022. tab, graf
Artigo em Inglês | LILACS | ID: biblio-1249271

RESUMO

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% 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.


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.


Assuntos
Escherichia coli/genética , alfa-Amilases/genética , alfa-Amilases/metabolismo , Temperatura , Estabilidade Enzimática , Clonagem Molecular , Geobacillus , Concentração de Íons de Hidrogênio
14.
Braz. j. biol ; 82: e249971, 2022. tab
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-1278485

RESUMO

Abstract Stunting is a significant public health problem in low- and middle-income countries. This study assessed the prevalence of stunting and associated risk factors of stunting among preschool and school-going children in flood-affected areas of Pakistan. A cross-sectional study was conducted by visiting 656 households through multi-stage sampling. Respondent's anthropometric measurements, socio-demographic information and sanitation facilities were explored. A logistic regression model was used to determine determinants of stunting, controlling for all possible confounders. The overall prevalence of stunting in children was 40.5%, among children 36.1% boys and 46.3% of girls were stunted. The prevalence of stunting in under-five children was 50.7%. Female children (OR=1.35, 95% CI:0.94-2.0), children aged 13-24 months (OR=6.5, 95% CI: 3.0-13.9), mothers aged 15-24 years (OR=4.4, 95% CI: 2.6-7.2), joint family (OR=2.1, 95% CI: 1.4-3.0) did not have access to improved drinking water (OR=3.3, 95% CI: 1.9-5.9), and the toilet facility (OR=2.8, 95% CI, 1.9-4.3), while the children from district Nowshera (OR=1.7, 95% CI: 0.9-3.2) were significantly (P<0.05) associated in univariate analysis. The regression model revealed that child age, maternal age, family type, quality of water, and toilet facility, were the significant (P<0.05) factors contributing to child stunting in the flood-hit areas. Identification of key factors might be helpful for policymakers in designing comprehensive community-based programs for the reduction of stunting in flood-affected areas. In disasters such as flood, the detrimental consequences of the stunting problem could be even more on children. Evidence-based education and care must be provided to the families in the flood-affected regions to reduce the stunting problem. The determinants of stunting should be targeted by making comprehensive policies regarding proper nutrition, livelihood, clean water, and sanitation facilities in flood-hit regions.


Resumo A baixa estatura é um problema significativo de saúde pública em países de baixa e média renda. Este estudo avaliou a prevalência de nanismo e os fatores de risco associados de nanismo entre crianças em idade pré-escolar e em idade escolar em áreas afetadas por inundações do Paquistão. Foi realizado um estudo transversal visitando 656 domicílios por meio de amostragem em múltiplos estágios. As medidas antropométricas do entrevistado, informações sociodemográficas e instalações de saneamento foram exploradas. Um modelo de regressão logística foi usado para determinar os determinantes do nanismo, controlando todos os possíveis fatores de confusão. A prevalência geral de baixa estatura em crianças foi de 40,5%, entre as crianças 36,1% dos meninos e 46,3% das meninas com baixa estatura. A prevalência de baixa estatura em crianças menores de 5 anos foi de 50,7%. Crianças do sexo feminino (OR = 1,35, IC de 95%: 0,94-2,0), crianças de 13-24 meses (OR = 6,5, IC de 95%: 3,0-13,9), mães de 15-24 anos (OR = 4,4, IC de 95%: 2,6-7,2), família conjunta (OR = 2,1, IC 95%: 1,4-3,0) não tiveram acesso a água potável de qualidade (OR = 3,3, IC 95%: 1,9-5,9) e a banheiro (OR = 2,8, IC de 95%, 1,9-4,3), enquanto as crianças do distrito de Nowshera (OR = 1,7, IC de 95%: 0,9-3,2) foram significativamente (P < 0,05) associadas na análise univariada. O modelo de regressão revelou que a idade da criança, idade materna, tipo de família, qualidade da água e banheiro foram os fatores significativos (P < 0,05) que contribuíram para a baixa estatura infantil nas áreas afetadas pelas enchentes. A identificação de fatores-chave pode ser útil para os formuladores de políticas no planejamento de programas comunitários abrangentes para a redução da baixa estatura em áreas afetadas pelas enchentes. Em desastres como enchentes, as consequências prejudiciais do problema de baixa estatura podem ser ainda maiores para as crianças. Educação baseada em evidências e cuidados deve ser fornecida às famílias nas regiões afetadas pelas enchentes para reduzir o problema de nanismo. Os determinantes do retardo de crescimento devem ser almejados pela formulação de políticas abrangentes sobre nutrição adequada, meios de subsistência, água potável e instalações de saneamento nas regiões afetadas pelas enchentes.


Assuntos
Humanos , Masculino , Feminino , Pré-Escolar , Criança , Inundações , Transtornos do Crescimento/epidemiologia , Paquistão/epidemiologia , Instituições Acadêmicas , Prevalência , Estudos Transversais , Fatores de Risco
15.
Braz J Biol ; 83: e248281, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34852158

RESUMO

The COVID-19 is a contagious viral disease, was first emerged in Wuhan, China in December 2019 and became the whole world on alert. The mortality rate in top most countries in Asia with special reference to Pakistan has been focused. Since February 26 to September 2020 the total confirmed cases and mortality rate was measured through Wikipedia and the notable journals. Iran is the only country having highest number of deaths (5.73%) followed by Indonesia (3.77%) while Saudi Arabia shows the lowest number of deaths as 1.39%. In Pakistan the first case was confirmed in 26th February, 2020. The nCov-19 has closely related to severe acute respiratory syndrome (SARS) hence SARS COV-2 was named. This virus is responsible for more than 33.9 million deaths in over all the world as of 20th September, 2020. The number of new cases is increasing time to time. Sindh province of Pakistan has reported the highest number of cases till September, 20, 2020 as compared to other parts of the country and has the highest number of death followed by Khyber Pakhtunkhwa. Because of the person to person contact the disease is spreading rapidly. The individuals who has already infected with other diseases like cancer or diabetic etc. are vulnerable. The nCOV-19 is the most contagious due to its mode of transmission. There is still no vaccine is available for the treatment of disease caused by nCoV-2019. It is therefore the only option to control this pandemic is to adopt effective preventive measures.


Assuntos
COVID-19 , Pandemias , China , Humanos , Paquistão/epidemiologia , SARS-CoV-2
16.
Trials ; 22(1): 792, 2021 Nov 11.
Artigo em Inglês | MEDLINE | ID: mdl-34763711

RESUMO

BACKGROUND: Alcohol consumption causes a spectrum of liver abnormalities and leads to over 3 million deaths per year. Alcoholic hepatitis (AH) is a florid presentation of alcoholic liver disease characterized by liver failure in the context of recent and heavy alcohol consumption. The aim of this study is to explore the potential benefits of the IL-1ß antibody, canakinumab, in the treatment of AH. METHODS: This is a multicentre, double-blind, randomised placebo-controlled trial. Participants will be diagnosed with AH using clinical criteria. Liver biopsy will then confirm that all histological features of AH are present. Up to 58 participants will be recruited into two groups from 15 centres in the UK. Patients will receive an infusion of Canakinumab or matched placebo by random 1:1 allocation. The primary outcome is the difference between groups in the proportion of patients demonstrating histological improvement and will compare histological appearances at baseline with appearances at 28 days to assign a category of "improved" or "not improved". Patients with evidence of ongoing disease activity will receive a second infusion of canakinumab or placebo. Participants will be followed up for 90 days. Secondary outcomes include mortality and change in MELD score at 90 days. DISCUSSION: This phase II study will explore the benefits of the IL-1ß antibody, canakinumab, in the treatment of AH to provide proof of concept that inhibition of IL-1ß signalling may improve histology and survival for patients with AH. TRIAL REGISTRATION: EudraCT 2017-003724-79 . Prospectively registered on 13 April 2018.


Assuntos
Hepatite Alcoólica , Anticorpos Monoclonais Humanizados , Ensaios Clínicos Fase II como Assunto , Método Duplo-Cego , Hepatite Alcoólica/diagnóstico , Hepatite Alcoólica/tratamento farmacológico , Humanos , Estudos Multicêntricos como Assunto , Ensaios Clínicos Controlados Aleatórios como Assunto , Resultado do Tratamento
17.
Mymensingh Med J ; 30(4): 954-959, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34605462

RESUMO

Biocides, including disinfectants and antiseptics, are used for a variety of topical and hard surface applications in health care facilities. Biocides play a significant role for preventing and controlling nosocomial infections. However, failures in the antimicrobial activities of biocides have been reported. The resistance mechanism to disinfectants is usually determined by genes which are related to resistance to quaternary ammonium compounds, namely, qacE, qacΔE1 that are found in Gram-negative bacteria. The aim of this study is to detect the prevalence of Biocides resistance genes, qacE and qacΔE1, in clinical isolates of Pseudomonas spp. It was carried out from March 2017 to July 2018 in the department of Microbiology, Mymensingh Medical College, Mymensingh, Bangladesh. Samples were collected from Outpatient of ENT department, MMCH. In this study, 300 clinical samples of CSOM cases were tested by the PCR method. The present study shows detection of biocide resistance genes (qacE, qacΔE1) among 87 isolated Pseudomonas spp by uniplex PCR. Among 72 clinical isolates of Pseudomonas aeruginosa 67(93.05%) had the gene qacEΔ1 and 25(34.72%) had the gene qacE. In addition other 15 Pseudomonas spp 3(20%) isolates had the qacEΔ1 gene and 2(13.33%) isolates had the qacE gene. In this study there is a marked difference in detection of the qacEΔ1 gene between the MDR and non MDR P. aeruginosa isolates. The qacEΔ1 was identified in 50 of 54(92.59%) MDR isolates and 7 of 18(38.89%) non MDR strains respectively. While gene qacE was detect 25(46.29%) MDR isolates and did not show any qacEΔ1gene in non MDR isolates. This study shows that the genes, qacE, qacΔE1 are widespread among Pseudomonas aeruginosa, they are higher in MDR strains than non MDR strains.


Assuntos
Desinfetantes , Antibacterianos/farmacologia , Bangladesh/epidemiologia , Hospitais , Humanos , Testes de Sensibilidade Microbiana , Pseudomonas/genética
18.
Braz J Biol ; 83: e247890, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34468530

RESUMO

Asiatic black bear is present in variety of habitats like broad-leaves and coniferous forests, extending form sea level to 4300m elevation and change their habitat for food purpose seasonally. The present study was conducted at Kaghan and Siran Valleys, District Mansehra, Khyber Pakhtunkhwa, Pakistan to assess habitat of black bear. Line transect method was used for observation of bear signs. Twelve meter circular radius plots were selected for the concern vegetation's i.e. (trees, shrubs and herbs) and three to six plots were placed in each transect. At the result of sign survey, thirteen different categories of bear signs were recorded and encounter rate was calculated for each sign. A total of 1858 signs were observed during field surveys. Total (81%) coniferous species were recoded among trees, with the highest appearance of Pinus wallichiana (34.22%) and Spruce spp (27.76%), similarly broad leaves trees (18.56%) were also recoded from habitat plots. Most of the signs were encountered in bushy areas, whereas high number of Viburnum Spp (60.29%) was present. It is indicated that black bear prefers blend of Coniferous Trees, Viburnum and Ferns Species; probably because these plants provide enough food, protection, and meticulous shelter because more than 80% of habitat composed of these three species. Currently habitat destruction and increase in human population are the up-growing issues for wild animals (especially Asiatic black bear), which is highly sensitive to such problems. High levels of conservation efforts are recommended for the protection of black bear habitat and to avoid human interference in their territory.


Assuntos
Ursidae , Acetilcisteína , Animais , Ecossistema , Humanos , Paquistão , Árvores
19.
J Neuromuscul Dis ; 8(6): 899-926, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34511509

RESUMO

There are growing numbers of adults with Duchenne Muscular Dystrophy living well into their fourth decade. These patients have complex medical needs that to date have not been addressed in the International standards of care. We sought to create a consensus based standard of care through a series of multi-disciplinary workshops with specialists from a wide range of clinical areas: Neurology, Cardiology, Respiratory Medicine, Gastroenterology, Endocrinology, Palliative Care Medicine, Rehabilitation, Renal, Anaesthetics and Clinical Psychology. Detailed reports of evidence reviewed and the consensus building process were produced following each workshop and condensed into this final document which was approved by all members of the Adult North Star Network including service users. The aim of this document is to provide a framework to improve clinical services and multi-disciplinary care for adults living with Duchenne Muscular Dystrophy.


Assuntos
Consenso , Distrofia Muscular de Duchenne/terapia , Padrão de Cuidado , Adulto , Humanos , Inquéritos e Questionários
20.
Braz J Biol ; 83: e250179, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34524376

RESUMO

Diabetes mellitus (DM) is a non-communicable disease throughout the world in which there is persistently high blood glucose level from the normal range. The diabetes and insulin resistance are mainly responsible for the morbidities and mortalities of humans in the world. This disease is mainly regulated by various enzymes and hormones among which Glycogen synthase kinase-3 (GSK-3) is a principle enzyme and insulin is the key hormone regulating it. The GSK-3, that is the key enzyme is normally showing its actions by various mechanisms that include its phosphorylation, formation of protein complexes, and other cellular distribution and thus it control and directly affects cellular morphology, its growth, mobility and apoptosis of the cell. Disturbances in the action of GSK-3 enzyme may leads to various disease conditions that include insulin resistance leading to diabetes, neurological disease like Alzheimer's disease and cancer. Fluoroquinolones are the most common class of drugs that shows dysglycemic effects via interacting with GSK-3 enzyme. Therefore, it is the need of the day to properly understand functions and mechanisms of GSK-3, especially its role in glucose homeostasis via effects on glycogen synthase.


Assuntos
Diabetes Mellitus , Resistência à Insulina , Glucose , Quinase 3 da Glicogênio Sintase , Homeostase , Humanos
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