Designing and Expression of Recombinant Chimeric Spike Protein from SARS-CoV-2 in Escherichia coli and Its Immunogenicity Assessment.

Since December 2019, the world has been grappling with an ongoing global COVID-19 pandemic. Various virus variants have emerged over the past two years, each posing a greater threat than its predecessors. The recent appearance of the omicron variant (B.1.1.529) has raised significant alarm within the field of epidemiology due to its highly contagious nature and rapid transmission rate. The omicron variant possessed mutations in the key receptor-binding domain (RBD) region, the S region, and these modifications have shown a notable impact on the strain's susceptibility to neutralizing antibodies. Developing safe and efficient vaccines to prevent a future severe acute respiratory outbreak of coronavirus syndrome 2 (SARS-CoV-2) is significant. Viral surface spike proteins are ideal targets for vaccines. This study aimed to find a multi-subunit chimeric vaccine. After conducting bioinformatics analysis, the recombinant spike (RS) protein of SARS-CoV-2 was deliberately designed and subsequently produced using E. coli expression systems. The immunogenicity of RS and neutralizing antibody responses were evaluated on immunized BALB/c mice. There was a significant difference in antibody titers between RS-immunized mice and control groups. The endpoint of the serum antibody titer of mice immunized with our chimeric protein was 2.5 times higher than that of the negative control. The chimeric construct could present multiple antigens simultaneously, influentially affecting immunization. Sera from mice vaccinated by RS could recognize the SARS-CoV-2 virus and neutralize antibodies. Our chimeric peptide could bind to antibodies in the serum of patients infected with different serotypes of the SARS-CoV-2 virus, such as alpha, delta, and omicron variants. The results indicated that the RS protein would be a potential novel antigenic candidate for subunit vaccine development and could be used as a useful alternative to generate diagnostic serological tests for SARS-CoV-2 infection.

Antigenicity and immunogenicity of SARS-CoV-2 surface glycoprotein fragment in CHO cells.

Background and Objectives: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike (S) glycoprotein that projects from the virus surface is highly immunogenic. It is considered to be the target of many neutralizing antibodies as well as a target in vaccine design efforts. Evaluation the immunogenicity of a recombinant fragment of the spike protein (rfsp) that is comprised of Receptor Binding Domain (RBD), S1/S2 cleavage site, and fusion peptide (FP) as immunogenic proteins of SARS-COV-2, in BALB/c mice and evaluation of the efficacy of epitopes rfsp as a multi-subunit chimeric vaccine. Materials and Methods: The present study made use of CHO-K1 (Chinese hamster ovary K1) cells to create a cell line for constant expression rfsp. The rfsp was purified with Ni-NTA chromatography and confirmed by Western blotting. The immunogenicity and neutralizing antibody efficacy of rfsp were evaluated in BALB/c mice. ELISA was employed to test rfsp via sera of COVID-19 convalescent patients infected with SARS-CoV-2 alpha and delta variants. Results: Our results showed significant differences in antibody titers in immunized mice compared to the control groups and neutralizing antibodies were positive, sera from mice immunized are capable of bound SARS-CoV-2 virus, chimer peptide is capable bound antibodies patients infected with SARS-CoV-2 and patients infected with delta variant SARS-CoV-2. Conclusion: Overall, these results indicate that rfsp protein would be a novel potential antigen candidate for the development of a subunit SARS CoV-2 vaccine and rfsp has the potential to be a useful option for the development of the assays for serodiagnosis of SARS-CoV-2 infection.

Performance Challenges in a Newly Established Clean Room at Omid Hospital, Isfahan, Iran: A Descriptive Study.

Objective: This study aimed to comprehensively assess the challenges faced by a newly established clean room in the oncology center of Omid Hospital, Isfahan, Iran, one of the first of its kind in the country. The research also sought to identify the underlying causes of these challenges and propose potential solutions to address them. Methods: A 6-month cross-sectional study was conducted from December 2021 to May 2022. International guidelines such as British Columbia Cancer Agencies' guideline of hazardous drugs, the National Institute for Occupational Safety and Health guideline for working with hazardous drugs, and United States pharmacopeia related to cleanroom performance were studied, translated, and summarized into a checklist. The staff performance in Omid Hospital's clean room was compared to the data collection form, and all medication errors were documented and analyzed. The study also explained the underlying causes of these challenges and proposed potential solutions. Findings: Among 1005 chemotherapy regimens, 836 errors were detected, stemming from issues such as engineering and construction challenges, lack of human resources and essential equipment, and budgetary constraints. Conclusion: Despite the involvement of a trained oncology clinical pharmacist, Omid Hospital's cleanroom faces significant challenges within the medical and hospital system, leading to non-standard challenges. The study recommends multidisciplinary approaches in the hospital to mitigate these challenges and improve cleanroom performance.

Antagonistic effect of isolated probiotic bacteria from natural sources against intestinal Escherichia coli pathotypes.

BACKGROUND: Probiotics are live microorganisms which are beneficial bacteria that are normal flora of the digestive system which, in determined amounts, show beneficial effects on host health, and can balance gastrointestinal microflora. Digestive tract diseases such as diarrhea are one of the major causes of child mortality in developing countries. Different pathotypes of Escherichia coli cause diarrhea that affects children, therefore reduction of these colonization strains in humans or animals can decline gastrointestinal disorders such as diarrhea. OBJECTIVE: The aim of this study was to determine the antimicrobial effect of probiotic bacterial strains isolated from different natural sources against 4 pathotypes of pathogenic E. coli using disk and well diffusion methods. METHODS: This cross-sectional study was conducted from December 2013 to July 2014 on Martyr Chamran University in Ahwaz city. A total of 13 probiotic colonies isolated from 20 samples of traditional dairy products including yogurt, cheese and milk, and 20 samples of vegetables including carrots and cabbages (red and white), of which 5 isolates were selected to determine the antimicrobial effect against 4 Escherichia coli pathotypes, randomly. The antimicrobial effect was evaluated using two methods: disk diffusion and well diffusion tests and measuring growth inhibition zones of probiotics against 4 pathotypes of pathogenic E. coli. RESULTS: Obtained results showed growth inhibition effects of all 5 probiotic strains against Escherichia coli pathotypes in both used methods. But in comparison Lactobacillus plantarum had higher growth inhibitory effects in both methods. CONCLUSION: results of this study demonstrated high antimicrobial effect of probiotic bacteria against pathogenic Escherichia coli strains. It indicated a positive and beneficial role of probiotics in human health and prevention of illness.

The antimicrobial activity of probiotic bacteria Escherichia coli isolated from different natural sources against hemorrhagic E. coli O157:H7.

BACKGROUND: Diarrheal diseases have been seen in all geographical areas throughout the world. Therefore, considering treatment, could be deemed a necessary action. OBJECTIVE: The aim of this study was to determine the antimicrobial effect of probiotic bacterial strains isolated from different natural sources against 2 pathotypes of pathogenic E. coli. METHODS: This cross-sectional study of Martyr Chamran University of Ahvaz was carried out from December 2013 to July 2014. A total of 13 probiotic colonies isolated from 20 samples of traditional dairy products including (yogurt, cheese, milk) and 20 samples of vegetables including carrots and cabbages (red and white) of which 5 isolates were selected to evaluate the antimicrobial effect against 2 Escherichia coli pathotypes, randomly. Antimicrobial effect was evaluated using two methods: disk diffusion and well diffusion tests and measuring growth inhibition zones of probiotics against 2 pathotypes of pathogenic E. coli. RESULTS: Obtained results showed growth inhibition effects of all 5 probiotic strains against Escherichia coli pathotypes in both used methods. All selected strains showed considerable antimicrobial effect on Escherichia coli O157:H7 strain, but had no inhibitory effect against Enterohemorrhagic Escherichia coli. CONCLUSION: This study demonstrated considerable antimicrobial effect against E. coli O157:H7 strain. Due to this, characteristic and similar antimicrobial effects of probiotics bacteria, increasing use of the probiotics as a natural and modern method for prevention of different diseases is recommended.

Evaluation of a Therapeutic Interchange from Fluticasone/Salmeterol to Mometasone/Formoterol in Patients with Chronic Obstructive Pulmonary Disease.

BACKGROUND: Combination treatment with an inhaled corticosteroid and long-acting beta2-agonist is among the many treatment options for chronic obstructive pulmonary disease (COPD) that has been shown to improve clinical outcomes. While mometasone/formoterol does not currently have an FDA-approved indication for COPD, evidence from 2 phase 3 trials demonstrated that mometasone/formoterol can improve lung function and was well tolerated in patients with moderate-to-very severe COPD. Based on these data, a therapeutic interchange was implemented in the Kaiser Permanente Mid-Atlantic States region to convert patients with a COPD diagnosis from fluticasone/salmeterol to mometasone/formoterol. OBJECTIVE: To evaluate the impact of a therapeutic interchange from fluticasone/salmeterol to mometasone/formoterol on health outcomes in patients with COPD in a large ambulatory and managed care setting. METHODS: The investigators retrospectively reviewed the electronic medical records of patients with a COPD diagnosis who had a prescription for fluticasone/salmeterol converted to mometasone/formoterol between March 6, 2011, to March 6, 2013. Kaiser Permanente's Pharmacy and Therapeutics Committee provided recommended equivalent doses for conversion from fluticasone/salmeterol to mometasone/formoterol. Nonetheless, the final approval for the change in medication and selection of the dose was left to each physician's clinical judgment. Patients were excluded if they were (a) prescribed fluticasone/salmeterol 100/50 mcg, which has no equivalent mometasone/formoterol dose; (b) less than aged 18 years; or (c) prescribed fluticasone/salmeterol for a duration of less than 6 months preconversion to mometasone/formoterol. In addition, patients who left the Kaiser Permanente network or became deceased during the study period of interest were excluded. After the application of the inclusion and exclusion criteria, 521 patients were included in the data analysis. The primary endpoint was the determination of the difference in the occurrence of COPD exacerbations 6 months pre- and postconversion from fluticasone/salmeterol to mometasone/formoterol. COPD exacerbations were defined by the diagnosis or documentation of a COPD exacerbation during any hospitalizations, urgent care (UC)/emergency department (ED) visits, or clinic encounters. Secondary outcomes included the determination of the difference in the occurrence of intensive care unit admissions, hospitalizations, UC/ED visits, and clinic encounters for COPD exacerbations 6 months pre- and postconversion; number of patients who required modification in therapy; and any reasons for mometasone/for-moterol discontinuation postconversion. Patients served as their own controls to compare any differences in outcomes while taking mometasone/formoterol versus fluticasone/salmeterol. RESULTS: Within our patient population, 34.2% (n = 178) of patients experienced at least 1 COPD exacerbation while prescribed fluticasone/salmeterol compared with 28.6% (n = 149) of patients while prescribed mometasone/formoterol (P = 0.030). Mometasone/formoterol therapy did not demonstrate any statistically significant differences in the secondary outcomes (P < 0.050). A later subgroup analysis of the primary outcome revealed that factors associated with a statistically significant decrease in the occurrence of COPD exacerbations were male sex (P = 0.023), comorbid asthma (P = 0.026), and conversion from fluticasone/salmeterol to a more potent dose of mometasone/formoterol (P = 0.014). CONCLUSIONS: There was a statistically significant decrease in the proportion of patients who experienced COPD exacerbations postconversion from fluticasone/salmeterol to mometasone/formoterol. This study is an example of a real-world therapeutic interchange that provides additional data to support the use of mometasone/formoterol for its unlabeled COPD indication. DISCLOSURES: No outside funding supported this study. The authors report no financial or other conflicts of interest related to the subject of this article. All authors contributed to study design and manuscript revision. Yip collected and analyzed data and prepared the manuscript.

Anticholinergic burden: clinical implications for seniors and strategies for clinicians.

OBJECTIVE: To review the current literature assessing the risks associated with the use of anticholinergic (AC) medications in older adults and to provide recommendations for pharmacists to incorporate the evaluation of AC burden as a component of a medication therapy management program for older adults. DATA SOURCES: A MEDLINE/PubMed search was conducted from January 1990 to July 2011 using the terms anticholinergic, antimuscarinic, geriatric, aged, and elderly. References cited in studies and reviews identified in this search were also evaluated. Articles published in languages other than English or conducted in nonhuman species were not evaluated. STUDY SELECTION: Studies and reviews were included if they evaluated adverse events associated with the use of AC medications in the older adult population. DATA EXTRACTION: Data were extracted by the method of independent extraction by multiple observers based on their selection of sections. DATA SYNTHESIS: The review presents evidence that AC medications are associated with negative outcomes in older adults and the importance of health care provider interventions to avoid these consequences. CONCLUSION: AC burden is associated with adverse drug events and negative health outcomes in older adults. Health care providers should carefully assess the risks versus benefits of using medications with AC properties to minimize AC burden and prevent adverse outcomes in this vulnerable patient population. AC burden should be considered as acomponent of a medication therapy management program for seniors.

Results of a safety initiative for patients on concomitant amiodarone and simvastatin therapy in a Veterans Affairs medical center.

BACKGROUND: The FDA revised the labels of amiodarone and simvastatin in 2002 to warn of increased risk of rhabdomyolysis, the most severe form of myopathy, when the 2 drugs are taken concomitantly in doses greater than 20 milligrams (mg) per day of simvastatin. The FDA reissued the warning in 2008 after receiving reports of 52 cases of rhabdomyolysis in the Adverse Event Reporting System (AERS) after the label changes in 2002 and suggested use of an alternative statin for patients receiving amiodarone who require more than 20 mg simvastatin to attain lipid goals. OBJECTIVES: To (a) assess the prevalence of concomitant amiodarone and simvastatin in doses greater than 20 mg per day and the frequency of additional risk factors for myopathy in these patients, and (b) implement and evaluate a protocol to convert patients receiving this combination to alternative statins. METHODS: A review was conducted of all patients with active prescriptions for both simvastatin at doses greater than 20 mg per day and amiodarone from a Veterans Affairs (VA) medical center as of November 1, 2008. Data collected included demographics, duration of therapy, baseline lipid and aminotransferase values, and risk factors for myopathy (i.e., aged 80 years or older; female sex; small body frame; hypothyroidism; hepatic or renal insufficiency; diabetes; alcohol abuse; and use of medications, such as gemfibrozil and nicotinic acid, that may increase the risk of myopathy). Patients were converted to either pravastatin or rosuvastatin based on baseline simvastatin dose, low-density lipoprotein cholesterol (LDL-C) level, and renal function. The conversion protocol was developed to maintain LDL-C lowering with potentially safer statins. Follow-up lipid and aminotransferase values were collected as customary clinical markers of efficacy and safety, respectively, for patients converted per the protocol. Because creatine kinase values are not routinely assessed in clinical practice, they were not available as part of the current protocol. RESULTS: Of the 48,612 patients who accessed the pharmacy in this VA medical center, 17,760 (36.5%) had an active order for simvastatin 40 mg or 80 mg per day, and 92 of these patients (0.52%) also had an active order for amiodarone. These patients were prescribed simvastatin primarily for secondary prevention (88 [95.7%] with coronary artery disease [CAD] as the indication for statin therapy), and were highly controlled with mean (SD) baseline LDL-C of 71 (21) mg per deciliter. The mean (median) duration of therapy on the combination of amiodarone plus simvastatin 40 mg or more per day was 43 (37) months. Of the 92 patients, 26 (28.3%), 35 (38.0%), and 18 (19.6%) patients had 1, 2, or 3 or more additional risk factors for myopathy, respectively. 16 patients were not converted per protocol to an alternate statin (4 were taken off amiodarone, 2 were taken off statin therapy, 6 had the simvastatin dose reduced to 20 mg per day or less, and 4 were converted to an alternate statin off-protocol), and 14 patients did not have follow-up laboratory values. For the 62 patients converted per protocol and with follow-up laboratory values, there were no statistically significant changes in mean lipid or aminotransferase values after conversion. One patient reported symptoms of myalgia after conversion to rosuvastatin; however, the conversion protocol did not require obtaining creatine kinase values. CONCLUSION: 0.19% of patients (n=92) with pharmacy dispensing records in this VA facility in 2008 received amiodarone in combination with simvastatin in doses greater than 20 mg per day, and the majority of patients had additional risk factors for myopathy. There were no significant changes in mean laboratory values for lipids or aminotransferase for the 62 patients (67%) who were converted and who had baseline and follow-up values; there was 1 case of self-reported myalgia in a patient converted to rosuvastatin.