Introduction: The outbreak of novel coronavirus COVID-19 infections that started in China late 2019 has spread rapidly and cases have been recorded worldwide. So, in this study, we sought clarification of the clinical characteristics and importance of changing the lymphocyte group, antibodies, CD markers, and interleukin-6 in the serum of COVID-19 patients, which may help to clarify the pathogen and develop new biomarkers. Material and Methods: Venous blood samples had been accumulated from patients before taking any medications. Sera had been separated and saved at (-20°C) until analysis. Serum anti-SARS-CoV-2 immunoglobulins (IgG, IgA, and IgM) were determined in plasma samples using enzyme-linked immunosorbent assays (ELISA) and Serum IL-6 was assessed. Results: Median IgM (p=0.001), IgG (p<0.0001), and IgA (p<0.001), were decreased in patients comparing with control the control group. There is a significant decrease in CD3+ and CD4+ cells compared to healthy individuals in patients infected with COVID-19 (p<0.0001). CD19+ cell count decreased in COVID-19 patients compared to that of the control group (p<0.0001). After calculating CD4+/CD8+ cell ratio decreased in COVID-19 patients (p<0.0001). However, CD56+ cells were found to be increased (p<0.0001). Conclusions: IgM, IgG, IgA levels and CD19+, CD4+ cells, CD4+/CD8+ cell ratio were found to be decreased whereas CD8+, CD3+, CD4+ cells were detected to be increased in COVID-19 patients compared to those of healthy controls. Keywords: IL-6, COVID-19, IgG, IgA, IgM, CD markers
Bioethanol produced from lignocellulose feedstock is a renewable substitute to declining fossil fuels. Pretreatment using ultrasound assisted alkaline was investigated to enhance the enzyme digestibility of waste paper. The pretreatment was conducted over a wide range of conditions including waste paper concentrations of 1-5%, reaction time of 10-30 min and temperatures of 30-70°C. The optimum conditions were 4 % substrate loading with 25 min treatment time at 60°C where maximum reducing sugar obtained was 1.89 g/L. Hydrolysis process was conducted with a crude cellulolytic enzymes produced by Cellulomonas uda (PTCC 1259).The maximum amount of sugar released and hydrolysis efficiency were 20.92 g/L and 78.4 %, respectively. Sugars released from waste paper were fermented into bioethanol with Saccharomyces cerevisiae. The maximum concentration of bioethanol estimated was 9.5 g/L after 48h of cultivation, the yield and volumetric productivity were 0.454 g/g glucose and 0.2g bioethanol/ L h. respectively. This study of ultrasound and sodium hydroxide treatment may be (we think) it will be a promising technique to develop bioethanol production from waste paper.
The main goal of the current study was cloning and expression of the human insulin gene in Pichia pastoris expression system, using genetic engineering techniques and its treatment application. Total RNA was purified from fresh normal human pancreatic tissue. RNA of good quality was chosen to obtain a first single strand cDNA. Human preproinsulin gene was amplified from cDNA strand, by using two sets of specific primers contain EcoR1 and Notl restriction sites. The amplified preproinsulin gene fragment was double digested with EcoRI and Not 1 restriction enzymes, then inserted into pPIC9K expression vector. The new pPIC9K-hpi constructive expression vector was transformed by the heat-shock method into the E.coli DH5α competent cells. pPic9k –hpi, which was propagated in the positive transformant E. coli cells, was isolated from cells and then linearised by restriction enzyme SalI, then transformed into Pichia pastoris GS115 using electroporation method. Genomic DNA of His+ transformants cell was extracted and used as a template for PCR analysis. The results showed, that the pPic9k – hpi was successfully integrated into the P. pastoris genome, for selected His+ transformants clones on the anticipated band at 330 bp, which is corresponded to the theoretical molecular size of the human insulin gene. To follow the insulin expression in transformans, Tricine–SDS gel electrophoresis and Western blot analysis were conducted. The results showed a successful expression of recombinant protein was detected by the presence of a single major band with about (5.8 KDa) on the gel. These bands correspond well with the size of human insulin with the theoretical molecular weight (5.8 KDa).
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