Escherichia coli as a versatile cell factory: Advances and challenges in recombinant protein production.

E. coli plays a substantial role in recombinant protein production. Its importance increased with the discovery of recombinant DNA technology and the subsequent production of the first recombinant insulin in E. coli. E. coli is a widely used and cost-effective host to produce recombinant proteins. It is also noteworthy that a significant portion of the approved therapeutic proteins have been produced in this organism. Despite these advantages, it has some disadvantages, such as toxicity and lack of eukaryotic post-translational modifications that can lead to the production of misfolded, insoluble, or dysfunctional proteins. This study focused on the challenges and engineering approaches for improved expression and solubility in recombinant protein production in E. coli. In this context, solution strategies such as strain and vector selection, codon usage, mRNA stability, expression conditions, translocation to the periplasmic region and addition of fusion tags in E. coli were discussed.

The Effect of Hot and Cold Water Applications on Restless Legs Syndrome Complaints and Sleep Quality of Pregnant Women: A Randomized Controlled Study.

Background: Pregnant women experience restless legs syndrome and intervention studies are needed to prevent this. This study evaluated the effect of hot and cold water applications on the severity of restless leg syndrome and sleep quality in pregnant women. Methods: It is a randomized controlled trial. The subjects comprised 80 (hot water group = 26, cold water group = 27, control group = 27) pregnant women. Pregnant women in intervention groups applied hot or cold water to their legs before going to bed for seven days. The control group did not do any application other than routine care. The data were collected with the Restless Legs Syndrome Diagnostic Form, Personal Information Form, Restless Leg Syndrome Severity Grading Scale, The Pittsburgh Sleep Quality Index, Application Satisfaction Form, and Post-Application Follow-up Chart. Results: While the Restless Legs Syndrome Severity Grading Scale and Pittsburgh Sleep Quality Index pre-test scores of the groups were similar (P > .05), the post-test scores were lower in the intervention groups than in the control group (P < .001). The effect of the applications started on the second day and the level of satisfaction with the application was similarly high in both groups (P < .05). During the follow-up, the syndrome severity scores of both the intervention groups were similar (P > .05), while the sleep quality of the cold group was better (P < .05) and most of the pregnant women did not need reapplication and they were satisfied with the application (P < .05). Conclusion: These safe interventions reduced the severity of restless legs syndrome in pregnant women and improved the quality of their sleep. Nurses can use these applications in pregnancy care, which begin to show effect in as little as two days.

A Silver Compound with Potential Applications as an Anticancer, Antibacterial, Antifungal, and Anti-Alzheimer's Agent.

This study investigated the biological activity of the silver coordination compound K-22. The IC50 values of K-22 on cancer cell lines range from 0.797 µg/mL to 3.426 µg/mL, indicating that K-22 might preferably inhibit A549, Saos-2, MCF-7, and HT-29 cell proliferation and thus have better therapeutic activity. Furthermore, K-22 stimulated apoptosis via up-regulation of the mRNA and protein expression level of Bax/Bcl-2 ratio in A549, Saos-2, MCF-7, and HT-29. K-22 exhibited antimicrobial activity against S. aureus, E. faecalis, K. pneumonia, P. aeruginosa, C. utilis, and C. albicans. Experimental results show that the compound has inhibitory potential with an IC50 value of 178.10 µM for the BChE (butyrylcholinesterase) enzyme, which has a vital role in the progression of Alzheimer's disease. As a result, compound K-22 exhibits a strong potential for medical use due to its anticancer, antibacterial, antifungal, and anti-Alzheimer properties.

Incontinence-Associated Dermatitis: Prevalence in Intensive Care Units and Knowledge, Attitudes, and Practices of Nurses.

BACKGROUND: Incontinence-associated dermatitis (IAD) is a major concern among hospitals, especially in intensive care units (ICUs). PURPOSE: To describe ICU nurses' knowledge, attitudes, and practices of IAD and to examine the relationships with IAD prevalence in the ICU setting. METHODS: A descriptive correlational design was used including the Knowledge, Attitudes and Practices of Incontinence-Associated Dermatitis Questionnaire and 1-month IAD prevalence data. RESULTS: The prevalence of IAD in ICUs was 6.89%. A positive correlation was found between IAD prevalence and nurses' IAD knowledge and attitudes. Nurses working in the ICU for more than 7 years, caring for patients at high risk for or having IAD, and thinking IAD-related nursing practices were sufficient had significantly higher IAD knowledge, attitudes, and practices. CONCLUSIONS: Findings indicate that ICU nurses learned about IAD through clinical experience and have inadequate training on IAD. A standardized evidence-based care protocol for IAD should be developed.

The Effect of the Flipped Classroom Model on Teaching Clinical Practice Skills.

INTRODUCTION: No evidence was found in the literature for the use of the flipped classroom model in teaching clinical practice skills in paramedics. The study aimed to determine the effect of the flipped classroom model in teaching clinical practice skills to paramedic program students. METHODS: The study was a single-center, randomized controlled, single-blind parallel-group study. The research was carried out with a university's first-year paramedic program students in the 2021 to 2022 academic year. The students were divided into groups by stratified sampling (intervention group = 21, control group = 21). Five clinical skills practices at various times were explained to the intervention group with the flipped classroom model and to the control group with the traditional lecture method. The research data were collected with the Introductory Characteristics Information Form, Checklists, Time Tracking Form, and Students' Questionnaire for Evaluating the flipped classroom model. RESULTS: Although the students in the intervention group received a higher total score from all 5 clinical practice skills than the students in the control group, this difference was not statistically significant (P > .05). However, although the time allocated for applications in the intervention group was 40 to 75 minutes longer than in the control group, most students stated that this model successfully taught skills and theory, increased their motivation during the application, and reduced application and exam anxiety. DISCUSSION: The flipped classroom model can be used as an alternative method to the traditional system in teaching clinical practice skills. This innovative educational approach can be recommended as a student-centered method in clinical skills teaching.

RESTLESS LEGS SYNDROME AND INSOMNIA FREQUENCY IN PATIENTS WITH PSORIASIS.

BACKGROUND: To assess the prevalence and severity of RLS in psoriasis patients and to investigate its effects on sleep and quality of life. METHODS: Seventy patients with psoriasis in Trakya University Medical Faculty Dermatology Department and also applied to Neurology Department in the same center and 70 volunteer controls were enrolled in the study. Severity of the Restless Legs Syndrome (RLS) was determined using International Restless Legs Syndrome Study Group (IRLSSG) criteria among the patients who have been diagnosed with RLS based on IRLSSG criteria. The presence of insomnia in patients was detected using International Classification of Sleep Disorders (ICSD-II) criteria. Additionally, to evaluate the severity of the disease and quality of life, Psoriasis Area Severity Index (PASI) and Dermatology Life Quality Index (DLQI) tests were applied to the patients with psoriasis. RESULTS: RLS frequency in patients with psoriasis was 28 (40%) compared to the control group 10 (14.2%), and the difference was statistically significant (p<0.001). IRLSSG severity scores were higher in patients with psoriasis who had insomnia secondary to RLS compared to those who did not have insomnia (p<0.001). The mean values of PASI were 7.54 +/- 6.52 in the presence of insomnia and 3.27 +/- 2.69 in the absence of insomnia. The difference was statistically significant (p<0.001). No significant difference was found in DLQI scores between patients with RLS and without RLS or between patients with and without insomnia (p>0.05). DISCUSSIONS: RLS frequency in patients with psoriasis was significantly higher than in the control group. In addition, we found that RLS is more common but RLS severity was mildly high in patients with psoriasis relative to controls. However, there was a correlation between the increase in severity of RLS with secondary insomnia patients who were diagnosed with RLS. It may be beneficial to consider RLS and insomnia for each patient diagnosed with psoriasis.

Molecular pathway of anticancer effect of next-generation HSP90 inhibitors XL-888 and Debio0932 in neuroblastoma cell line.

Neuroblastoma is a common nervous system tumor in childhood, and current treatments are not adequate. HSP90 is a molecular chaperone protein that plays a critical role in the regulation of cancer-related proteins. HSP90 inhibition may exert anticancer effects by targeting cancer-related processes such as tumor growth, cell proliferation, metastasis, and apoptosis. Therefore, HSP90 inhibition is a promising strategy in the treatment of various types of cancer, and the development of next-generation inhibitors could potentially lead to more effective and safer treatments. XL-888 and Debio0932 is a next-generation HSP90 inhibitor and can inhibit the correct folding and stabilization of client proteins that cancer-associated HSP90 helps to fold correctly. In this study, we aimed to investigate the comprehensive molecular pathways of the anticancer activity of XL-888 and Debio0932 in human neuroblastoma cells SH-SY5Y. The cytotoxic effects of XL-888 and Debio0932 on the neuroblastoma cell line SH-SY5Y cells were evaluated by MTT assay. Then, the effect of these HSP90 inhibitors on the expression of important genes in cancer was revealed by Quantitative Real Time Polymerase Chain Reaction (qRT-PCR) method. The qRT-PCR data were evaluated using Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) biological process tools. Finally, the effect of HSP90 inhibitors on HSP27, HSP70 and HSP90 protein expression was investigated by Western blotting analysis. The results revealed that XL-888 and Debio0932 had a role in regulating many cancer-related pathways such as migration, invasion, metastasis, angiogenesis, and apoptosis in SH-SY5Y cells. In conclusion, it shows that HSP90 inhibitors can be considered as a promising candidate in the treatment of neuroblastoma and resistance to chemotherapy.

Synergistic induction of apoptosis in liver cancer cells: exploring the combined potential of doxorubicin and XL-888.

Combination therapy has been frequently preferred in treating various types of cancer in recent years. Targeted cancer therapy has also become one of the remarkable treatment modalities. Therefore, the aim of the study to investigate its cytotoxic and apoptotic effects on liver cancer cell lines by combining the classical chemotherapeutic drug doxorubicin (DOX) and a targeted agent, the new generation HSP90 inhibitor XL-888. The molecular docking method was used to predict the binding conformation of XL-888 on the human Hsp90. The single and combined cytotoxic effects of DOX and XL-888 on liver cancer cell lines HepG2 and HUH-7 were determined by MTT assay. The effect of the combined use of two drugs was evaluated using Chou and Talalay method. The levels of apoptotic genes and heat shock proteins gene and protein expression levels were investigated by quantitative real-time polymerase chain reaction and western blotting, respectively. Molecular docking results showed that XL-888 selectively binds to the ATP binding pocket of HSP90 with an estimated free binding energy of - 7.8 kcal/mol. DOX and XL-888 and their combination showed dose-dependent cytotoxic effect. The combination of drugs showed a synergistic effect on both cell lines. The results revealed that the combination of DOX and XL-888 potently induced apoptosis in liver cancer cell lines rather than using drugs alone. The combined treatment of DOX and XL-888 demonstrated synergistic cytotoxic and apoptotic effects on liver cancer cell lines, presenting a promising approach for combination therapy in liver cancer.

Thiolated α-cyclodextrin: The likely smallest drug carrier providing enhanced cellular uptake and endosomal escape.

This study aimed to evaluate the effect of thiolated α-cyclodextrin (α-CD-SH) on the cellular uptake of its payload. For this purpose, α-CD was thiolated using phosphorous pentasulfide. Thiolated α-CD was characterized by FT-IR and 1H NMR spectroscopy, differential scanning calorimetry (DSC), and powder X-ray diffractometry (PXRD). Cytotoxicity of α-CD-SH was evaluated on Caco-2, HEK 293, and MC3T3 cells. Dilauryl fluorescein (DLF) and coumarin-6 (Cou) serving as surrogates for a pharmaceutical payload were incorporated in α-CD-SH, and cellular uptake was analyzed by flow cytometry and confocal microscopy. Endosomal escape was investigated by confocal microscopy and hemolysis assay. Results showed no cytotoxic effect within 3 h, while dose-dependent cytotoxicity was observed within 24 h. The cellular uptake of DLF and Cou was up to 20- and 11-fold enhanced by α-CD-SH compared to native α-CD, respectively. Furthermore, α-CD-SH provided an endosomal escape. According to these results, α-CD-SH is a promising carrier to shuttle drugs into the cytoplasm of target cells.

Unchanged serum adipokine concentrations in the setting of short-term thyroidectomy-induced hypothyroidism.

OBJECTIVE: To investigate short-term effects of thyroidectomy-induced hypothyroidism on leptin, adiponectin, and resistin concentrations in association with anthropometric data. METHODS: Thirty premenopausal women with euthyroid nodular goiter-mean age, 44.0 ± 11.6 years; mean body mass index (BMI), 28.6 ± 5.9 kg/m2; 13 obese, 7 overweight, and 10 normal weight subjects-scheduled for total thyroidectomy were included in the study. Serum leptin, adiponectin, resistin, free triiodothyronine, free thyroxine, thyroid-stimulating hormone, glucose, insulin, and C-reactive protein concentrations, lipid profile, and anthropometric variables were determined in the euthyroid state (preoperatively) and the hypothyroid state (postoperatively, with a thyroid-stimulating hormone concentration >30 mIU/L). RESULTS: Body weight, BMI, waist and hip circumferences, body fat mass, and serum lipid concentrations increased significantly after thyroidectomy. No significant difference was found between preoperative and postoperative serum leptin, adiponectin, and resistin concentrations. Fat tissue mass-corrected leptin, adiponectin, and resistin concentrations did not differ significantly between euthyroid and hypothyroid periods. Thyroid hormone concentrations showed no significant correlations with adipokine levels. CONCLUSION: Serum adipokine concentrations seem not to change significantly during short-term thyroidectomy-induced hypothyroidism despite significant increases in body weight, BMI, fat mass, and lipid concentrations.