Huh-7 cell line as an alternative cultural model for the production of human like erythropoietin (EPO)

Erythropoietin (EPO) is a glycoprotein hormone which is required to regulate the production of red blood cells. Deficiency of EPO is known to cause anemia in chronically infected renal patients and they require regular blood transfusion. Availability of recombinant EPO has eliminated the need for blood transfusion and now it is extensively used for the treatment of anemia. Glycosylation of erythropoietin is essential for its secretion, stability, protein conformation and biological activity. However, maintenance of human like glycosylation pattern during manufacturing of EPO is a major challenge in biotechnology. Currently, Chinese hamster ovary (CHO) cell line is used for the commercial production of erythropoietin but this cell line does not maintain glycosylation resembling human system. With the trend to eliminate non-human constituent from biopharmaceutical products, as a preliminary approach, we have investigated the potential of human hepatoma cell line (Huh-7) to produce recombinant EPO.Initially, the secretory signal and Kozak sequences was added before the EPO mature protein sequence using overlap extension PCR technique. PCR-amplified cDNA fragments of EPO was inserted into mammalian expression vector under the control of the cytomegalovirus (CMV) promoter and transiently expressed in CHO and Huh-7 cell lines. After RT-PCR analysis, ELISA and Western blotting was performed to verify the immunochemical properties of secreted EPO.Addition of secretory signal and Kozak sequence facilitated the extra-cellular secretion and enhanced the expression of EPO protein. Significant expression (P < 0.05) of EPO was observed in the medium from Huh-7 cell line.Huh-7 cell line has a great potential to produce glycosylated EPO, suggesting the use of this cell line to produce glycoproteins of the therapeutic importance resembling to the natural human system

Effect of ellagic acid on growth and physiology of canola (Brassica napus L.) under saline conditions

Salinity stress is limiting growth and productivity of plants in many areas of the world. Plants adopted different strategies to minimize the effect of salt stress. A pot experiment was conducted to investigate the morphological and physiological changes produced in Canola (Brassica napus) by exogenous application of ellagic acid (EA) under saline conditions. EA is an antioxidant, expected to reduce the effect of salinity stress. The seeds of two canola cultivars, Rainbow and Oscar, were soaked for 6 h with different concentrations of EA (0, 55 and 110 µg/ml). The soaked seeds were sown in small pots. Salt stress was imposed on the plants by applying NaCl solutions of different concentrations (0, 60 and 120 mM) and the duration of stress was for four weeks. Salinity stress reduced seed germination and disturbed the morphological and physiological attributes of B. napus. Application of EA as seed soaking reduced the effect of salinity and enhanced the growth of plants. Overall, we could confirm a significant role of EA by inducing salinity tolerance in B. napus

Clinically Meaningful Change in 6 Minute Walking Test and the Incremental Shuttle Walking Test following Coronary Artery Bypass Graft Surgery

The 6-min walk test (6MWT) and incremental shuttle walk test (ISWT) are widely used measures of exercise tolerance, which depict favorable performance characteristics in a variety of cardiac and pulmonary conditions. Both tests are valid and reliable method of assessing functional ability in cardiac rehabilitation population. Several studies have calculated the minimal clinically important difference (MCID) of these exercise tests in different populations. The current study aims to estimate MCID of 6MWT and ISWT in patients after Coronary artery bypass graft (CABG) surgery. In this descriptive observational study, nonprobability purposive sampling technique was used to assess 89 post CABG patients. The participants performed the 6MWT and ISWT along with vital monitoring on third, fifth and seventh post operative days. The data was with calculation of 6MWT and ISWT MCID through distribution and anchor-based methods. Results showed significant improvement (p < 0.001) in 6MWT as well as in ISWT after seven days of in-patient cardiac rehabilitation. The minimal detectable difference of 6MWT determined by the distribution-based method was 36.11 whereas MCID calculated by Anchor based method was 195 m. The minimal detectable difference of ISWT determined by the distribution-based method was 9.94 whereas MCID calculated by Anchor based method was 42.5 m. In conclusion our results will assist the future researchers and clinicians to interpret clinical trials as well as to observe the clinical course of post operative cardiac patients

Effect of ellagic acid on growth and physiology of canola (Brassica napus L.) under saline conditions

Salinity stress is limiting growth and productivity of plants in many areas of the world. Plants adopted different strategies to minimize the effect of salt stress. A pot experiment was conducted to investigate the morphological and physiological changes produced in Canola (Brassica napus) by exogenous application of ellagic acid (EA) under saline conditions. EA is an antioxidant, expected to reduce the effect of salinity stress. The seeds of two canola cultivars, Rainbow and Oscar, were soaked for 6 h with different concentrations of EA (0, 55 and 110 µg/ml). The soaked seeds were sown in small pots. Salt stress was imposed on the plants by applying NaCl solutions of different concentrations (0, 60 and 120 mM) and the duration of stress was for four weeks. Salinity stress reduced seed germination and disturbed the morphological and physiological attributes of B. napus. Application of EA as seed soaking reduced the effect of salinity and enhanced the growth of plants. Overall, we could confirm a significant role of EA by inducing salinity tolerance in B. napus.© 2017 The Author(s

Ni tolerance and its distinguished amelioration by chelating agents is reflected in root radius of B. napus cultivars

The negative effect of excess nickel (Ni) on plants is well investigated but there is only little information on its influence on root anatomy and a possible amelioration by chelating agents. In this study, we utilized light microscopy to observe anatomical changes in canola (Brassica napus) roots and investigated the element content by X-ray microanalysis. Ni-tolerant (Con-II) and Ni-sensitive cultivars (Oscar) were selected for this purpose. The plants were treated with 30 ppm NiSO4. Then, citric acid and ethylene-diamine-tetra-acetic acid (EDTA) (alone or in combination) were applied to observe the influence of chelating agents in metal stress amelioration. Ni treatment led to significant swelling of the roots in the Con-II variety as compared to the cultivar Oscar. Application of EDTA reduced the root radius of Con-II plants and this effect for Ni tolerance is discussed. According to X-ray microanalyses, Ni ions were more dispersed in the sensitive cultivar as indicated by metal adsorption to the cell wall. We investigate the hypothesis that an enhanced capacity of binding metals to the cell wall allows the plants to tolerate more heavy metals.© Springer-Verlag GmbH Austria, part of Springer Nature 201

Effects of pre-operative isolation on postoperative pulmonary complications after elective surgery: an international prospective cohort study an international prospective cohort study

We aimed to determine the impact of pre-operative isolation on postoperative pulmonary complications after elective surgery during the global SARS-CoV-2 pandemic. We performed an international prospective cohort study including patients undergoing elective surgery in October 2020. Isolation was defined as the period before surgery during which patients did not leave their house or receive visitors from outside their household. The primary outcome was postoperative pulmonary complications, adjusted in multivariable models for measured confounders. Pre-defined sub-group analyses were performed for the primary outcome. A total of 96,454 patients from 114 countries were included and overall, 26,948 (27.9%) patients isolated before surgery. Postoperative pulmonary complications were recorded in 1947 (2.0%) patients of which 227 (11.7%) were associated with SARS-CoV-2 infection. Patients who isolated pre-operatively were older, had more respiratory comorbidities and were more commonly from areas of high SARS-CoV-2 incidence and high-income countries. Although the overall rates of postoperative pulmonary complications were similar in those that isolated and those that did not (2.1% vs 2.0%, respectively), isolation was associated with higher rates of postoperative pulmonary complications after adjustment (adjusted OR 1.20, 95%CI 1.05–1.36, p = 0.005). Sensitivity analyses revealed no further differences when patients were categorised by: pre-operative testing; use of COVID-19-free pathways; or community SARS-CoV-2 prevalence. The rate of postoperative pulmonary complications increased with periods of isolation longer than 3 days, with an OR (95%CI) at 4–7 days or ≥ 8 days of 1.25 (1.04–1.48), p = 0.015 and 1.31 (1.11–1.55), p = 0.001, respectively. Isolation before elective surgery might be associated with a small but clinically important increased risk of postoperative pulmonary complications. Longer periods of isolation showed no reduction in the risk of postoperative pulmonary complications. These findings have significant implications for global provision of elective surgical care. We aimed to determine the impact of pre-operative isolation on postoperative pulmonary complications after elective surgery during the global SARS-CoV-2 pandemic. We performed an international prospective cohort study including patients undergoing elective surgery in October 2020. Isolation was defined as the period before surgery during which patients did not leave their house or receive visitors from outside their household. The primary outcome was postoperative pulmonary complications, adjusted in multivariable models for measured confounders. Pre-defined sub-group analyses were performed for the primary outcome. A total of 96,454 patients from 114 countries were included and overall, 26,948 (27.9%) patients isolated before surgery. Postoperative pulmonary complications were recorded in 1947 (2.0%) patients of which 227 (11.7%) were associated with SARS-CoV-2 infection. Patients who isolated pre-operatively were older, had more respiratory comorbidities and were more commonly from areas of high SARS-CoV-2 incidence and high-income countries. Although the overall rates of postoperative pulmonary complications were similar in those that isolated and those that did not (2.1% vs 2.0%, respectively), isolation was associated with higher rates of postoperative pulmonary complications after adjustment (adjusted OR 1.20, 95%CI 1.05–1.36, p = 0.005). Sensitivity analyses revealed no further differences when patients were categorised by: pre-operative testing; use of COVID-19-free pathways; or community SARS-CoV-2 prevalence. The rate of postoperative pulmonary complications increased with periods of isolation longer than 3 days, with an OR (95%CI) at 4–7 days or ≥ 8 days of 1.25 (1.04–1.48), p = 0.015 and 1.31 (1.11–1.55), p = 0.001, respectively. Isolation before elective surgery might be associated with a small but clinically important increased risk of postoperative pulmonary complications. Longer periods of isolation showed no reduction in the risk of postoperative pulmonary complications. These findings have significant implications for global provision of elective surgical care