Cloning, expression and purification of escherichia coli modified phytase

Introduction: Phytases are the class of phosphatases, which are capable of hydrolyzing phytic acid. Phytases with the phytate degradation are able to reduce or eliminate the harmful effects of phytate. Acidic and thermal stable phytase with high yield and purity by a relatively inexpensive system had extensive application. So, in this study, by modification in enzyme sequence, recombinant phytase production with the shorter length and high expression level was assessed. Materials and Methods: The phytase gene sequence was obtained from the NCBI database. After bioinformatics studies and doing the noted modification for increasing protein expression, gene proliferation was done by using PCR. E. coli BL21 (DE3) was used to express the protein. Protein purification was performed by Ni-NTA kit and finally, enzyme activity was assessed. Results: Phytase was successfully expressed and purified. Enzyme activity assay showed a significant activity. Conclusion: Produced recombinant phytase had high activity in spite of eliminating parts of the enzyme. © 2017, Semnan University of Medical Sciences. All rights reserved

Comparison of the effects of preoperative celecoxib and gabapentin on pain, functional recovery, and quality of life after total knee arthroplasty: A randomized controlled clinical trial

Background: Acute pain is one of the main complaints of patients after total knee arthroplasty (TKA), which causes delayed mobility, increased morphine consumption, and subsequently increased costs. Therefore, the present study was performed to evaluate the preventive effect of preoperative celecoxib and gabapentin on reducing patient pain as a primary outcome after TKA surgery. Materials and Methods: This randomized, double-blind controlled clinical trial was performed on 270 patients with osteoarthritis that were candidates for TKA surgery allocated into three groups. In the first group, 900 mg of gabapentin was administered orally on a daily basis for 3 days before surgery. In the second group, 200 mg of oral celecoxib was administered twice daily for 3 days before surgery. In the third group, oral placebo was administered twice daily for 3 days before the surgery. The patients' pain score and knee and its functional score were recoded. Results: The mean of reduction pain in gabapentin and celecoxib groups was significantly lower than that of the control group at 12, 24, and 48 h after surgery (P 0.05). Furthermore, the two medication groups were not significantly different in this regard (P > 0.05). In addition, the knee score in the gabapentin group with the means of 85.40 +/- 5.47 and the celecoxib group with the means of 87.03 +/- 3.97 were significantly higher than those of the control group with the means of 78.90 +/- 4.39 in the 1(st) month after the surgery (P < 0.001). Conclusion: According to the results of the present study, the preventive administration of gabapentin and celecoxib showed a significant and similar effectiveness on reducing patient pain after TKA surgery and on improving the KSS and quality of life scores

Cloning and High-Level Expression of the Enzymatic Region of Phytase in E. coli

Phytase is an important enzyme poses great nutritional significance in humans and monogastric animals diets. The phytase production yield using wild sources, including micro-organisms, plants, and animals is sorely low. Thus, recombinant expression of phytase has received increasing interest for achieving production rate. Escherichia coli is the most preferred host for expression of heterologous proteins but overexpression of recombinant phytase in E. coli, met with limited success due to the sequestration of the enzyme into inclusion bodies. In the present study, artificial phytases gene with excellent thermostability and activity were designed by detecting the enzymatic region of the E. coli phytase gene by employing bioinformatics tools. Then, the PCR amplified recombinant gene was expressed in E. coli and the active enzyme was recovered from inclusion bodies. Employing cysteine amino acid in the dialysis buffer succeed to the superior activity of the enzyme with a specific activity of 73.8 U/mg. The optimum temperature and pH for enzyme activity were determined at 60 °C and 4, respectively. The novel recombinant enzyme illustrated perfect thermostability up to 70 °C with maintenance 75 of its activity. The enzyme was stable at pH range of 2�10. Moreover, the effects of ions and chemical compounds on enzyme stability and activity were assessed. © 2019, Springer Nature B.V

The Comparison Between 3.5-and 4.5-mm T-plates for Management of the Patients with Schatzker Type II Tibial Plateau Fractures that Referred to our Emergency Department: A Clinical Trial

Background: Tibial plateau fractures have become more frequent in recent years. The most prevalent Schatzker classification is type II, which is a lateral tibial plateau fracture with depression. Our null hypothesis was that the 3.5 T-plate and the 4.5 T-plate have no difference in the management of patients with Schatzker type II tibial plateau fractures. Materials and Methods: The current study is a clinical trial that was conducted on patients with tibial plateau fractures. The Knee Society Score (KSS) was this study's main outcome. Tourniquet time (TT) and patient quality of life using the 36-item Short Form Survey Instrument (SF-36) were secondary goals of the outcomes measurement study. VAS measured pain. Among 176 patients, 89 and 87 of cases underwent surgical treatment with 3.5-mm (group A) and 4.5-mm (group B) T-plate, respectively. The data were entered into SPSS software (version 25, IBM Corporation, Armonk, NY) and analyzed. Results: In our study, we evaluated 176 patients with a mean age of 34.8 +/- 15.2 years. Functional and clinical KSS scores were similar between the two groups throughout follow-up (P > 0.05). Regarding the other variables of VAS, TT, SF-36 physical function, and SF-36 mental health, no significant difference was observed between the two groups, and the two groups had similar averages in terms of these indicators (P > 0.05). Conclusion: According to the results, both plates had the appropriate functional outcomes in patients with split depression tibial plateau fracture