RESUMEN
This study was carried out to express human epidermal growth factor (hEGF) in Pichia pastoris GS115. For this aim, the hEGF gene was cloned into the pPIC9K expression vector, and then integrated into P. pastoris by electroporation. ELISA-based assay showed that the amount of hEGF secreted into the medium can be affected by the fermentation conditions especially by culture medium, pH and temperature. The best medium for the optimal hEGF production was BMMY buffered at a pH range of 6.0 and 7.0. The highest amount of hEGF with an average yield of 2.27 µg/mL was obtained through an induction of the culture with 0.5% (v/v) methanol for 60 h. The artificial neural network (ANN) analysis revealed that changes in both pH and temperature significantly affected the hEGF production with the pH change had slightly higher impact on hEGF production than variations in the temperature.(AU)
Asunto(s)
Pichia , Factor de Crecimiento Epidérmico , Red Nerviosa , Ingeniería Genética , ADN RecombinanteRESUMEN
Abstract This study was carried out to express human epidermal growth factor (hEGF) in Pichia pastoris GS115. For this aim, the hEGF gene was cloned into the pPIC9K expression vector, and then integrated into P. pastoris by electroporation. ELISA-based assay showed that the amount of hEGF secreted into the medium can be affected by the fermentation conditions especially by culture medium, pH and temperature. The best medium for the optimal hEGF production was BMMY buffered at a pH range of 6.0 and 7.0. The highest amount of hEGF with an average yield of 2.27 µg/mL was obtained through an induction of the culture with 0.5% (v/v) methanol for 60 h. The artificial neural network (ANN) analysis revealed that changes in both pH and temperature significantly affected the hEGF production with the pH change had slightly higher impact on hEGF production than variations in the temperature.
Asunto(s)
Humanos , Pichia/metabolismo , Proteínas Recombinantes/metabolismo , Factor de Crecimiento Epidérmico/metabolismo , Pichia/genética , Proteínas Recombinantes/genética , Ensayo de Inmunoadsorción Enzimática , Expresión Génica , Clonación Molecular , Medios de Cultivo/química , Factor de Crecimiento Epidérmico/genética , Fermentación , Concentración de Iones de HidrógenoRESUMEN
This study was carried out to express human epidermal growth factor (hEGF) in Pichia pastoris GS115. For this aim, the hEGF gene was cloned into the pPIC9K expression vector, and then integrated into P. pastoris by electroporation. ELISA-based assay showed that the amount of hEGF secreted into the medium can be affected by the fermentation conditions especially by culture medium, pH and temperature. The best medium for the optimal hEGF production was BMMY buffered at a pH range of 6.0 and 7.0. The highest amount of hEGF with an average yield of 2.27µg/mL was obtained through an induction of the culture with 0.5% (v/v) methanol for 60h. The artificial neural network (ANN) analysis revealed that changes in both pH and temperature significantly affected the hEGF production with the pH change had slightly higher impact on hEGF production than variations in the temperature.
Asunto(s)
Factor de Crecimiento Epidérmico/metabolismo , Pichia/metabolismo , Proteínas Recombinantes/metabolismo , Clonación Molecular , Medios de Cultivo/química , Ensayo de Inmunoadsorción Enzimática , Factor de Crecimiento Epidérmico/genética , Fermentación , Expresión Génica , Humanos , Concentración de Iones de Hidrógeno , Pichia/genética , Proteínas Recombinantes/genéticaRESUMEN
Aim: The objective of this study is to compare the knowledge, practice and awareness of oral hygiene among three different age groups (15-30, 30-45 and 45-60-year-olds) of the Saudi population in Sakaka. Methods: 629 subjects aged between 15-60 years were randomly selected. Data were collected by self-administered structured questionnaires and analysis was done using the SPSS statistical software. The subjects were divided in to A group (e30 years), B group (d31 years - e45 years) and C group (d46 years). Results: 30.7% (A group) of subjects preferred commercial bush to clean their teeth (p<0.001), while 5.9% (B and C groups) choose Miswak for cleaning. Group B people preferred brushing only in the morning (24.5%), while 16.5% of group A respondents preferred brushing twice a day. 23.1% of group A subjects responded that brushing regularly was the ideal way to avoid caries (p<0.05). 51.8% of all age groups had no idea on the effects of fluoride on caries disease. The reason to visit dentist was mainly for extraction (30.5%) followed by pain relief (28.9%). 62.62% (p<0.05) subjects had carious lesions, whereas 56.6% subjects had fractured teeth. The use of tobacco (16.7%) was highly prevalent in group B subjects (p<0.05). Tea consumption among A group subjects (49.6%) more than other age groups (p<0.001). 81.2% subjected were free from gingival bleeding (p<0.05). Most of the people (48.6%) preferred frequent dental camps in response to their opinion to improve dental facilities in Sakaka, Al-Jouf. Conclusions: The present study shows that poorer knowledge and oral health conditions among 30-45-year-old subjects (B group) compared to the other two groups. Generally, these data may be important in the evaluation of the past and planning of future oral health prevention and treatment programs targeting the high-risk populations.