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Chemicals used for seed treatments help to increase the agricultural production by preventing pests and pathogens but also cause environmental and health problems. Thus, environmentally-friendly technologies need to be developed for a seed treatment that inactivates surface microflora and improves seed vigor. One such pulsed electric field (PEF) treatment applied to cucumber seeds in the range of 1.07-17.28 Joule (J) significantly enhanced a mean germination rate (MGR) by up to 9%, a normal seedling rate by 25.73%, and a resistance to 100 and 200 mM salt stresses by 96% and 91.67%, respectively, with a stronger and faster growth of roots and seedlings. PEF treatment provided 3.34 and 3.22 log-reductions in the surface microflora of total mold and yeast and total aerobic mesophilic bacteria, respectively. The electrical conductivity (EC) values of the control samples increased over time, from 4 to 24 h. Those of the PEF-treated samples after 4, 12, and 24th hours were also more affected by the measurement time not by the PEF treatment.

Finite element modeling has served as a cornerstone in understanding knee joint mechanics post-meniscectomy, yet the influence of varying knee geometries remains unknown. The present study aimed to fill that gap by employing statistical shape modeling to generate knee models from MRI data of 31 human knees, capturing the population's knee size and shape variations. Finite element simulations were conducted to replicate intact, partial, and total medial meniscectomy conditions during standing. The results revealed a substantial shift in load distribution from the medial to lateral compartment following medial meniscectomy with its magnitude depending on knee geometry. Cartilages experienced variable degrees of pressure changes at different sites, which could also be different for fluid and contact pressures. While changes in joint size led to somewhat predictable alterations in contact pressure, variations in joint shape resulted in unexpected changes in contact and fluid pressures, emphasizing the need for computational simulations. The average knee geometry exhibited the lowest contact and fluid pressures under the given loading and boundary conditions, in contrast to knees with shapes deviating from the average. This study highlights the significance of individual knee shape in the biomechanical outcome of meniscectomy, potentially explaining the variability in clinical outcomes observed post-surgery., Competing Interests: Declarations Competing interests The authors declare no competing interests. Ethical approval The use of MRI of human participants was approved by the University of Calgary, ethics ID#: REB15-1165. The MRI data used in the present study were obtained from the Osteoarthritis Initiative, https://nda.nih.gov/oai/. Dataset identifiers: 1200285; 1200816. No participant information is included in the manuscript., (© 2024. The Author(s).)

The purpose of this study was to assess the additional contribution of karyotyping compared with genome-wide non-invasive prenatal testing (NIPT) for pregnancies at intermediate risk for trisomy 21 (T21), calculated using the maternal serum screening without major structural anomalies detected through sonography. Karyotype results of all pregnancies undergoing invasive prenatal diagnostic testing between January 2013 and March 2022 were obtained from a large hospital-based laboratory. Pregnancies with no major structural anomalies on ultrasound (including soft markers) and an intermediate risk for T21 on maternal serum screening were included in this study. The additional contribution of karyotyping for abnormal karyotype results was calculated after excluding results that could theoretically be identified with genome-wide NIPT. Among the 511 pregnancies analyzed, 13 (2.54%) were found to have abnormal karyotype results, 9 (1.76%) of which could theoretically have been detected with genome-wide NIPT. Within the cohort, 6/263 (2.28%) of women aged 35 years and older, and 3/248 (1.20%) of women younger than 35 years had results that could have been detected with genome-wide NIPT. After excluding results detectable using genome-wide NIPT, the additional contribution of karyotyping was found as 4/502 (0.79%) for the entire cohort, 2/257 (0.77%) for women aged 35 years and older, 2/245 (0.81%) for women younger than 35 years. Of the 511 examined pregnancies at intermediate risk for T21 by maternal serum screening, genome-wide NIPT would have failed to detect 4 of 13 abnormal karyotype results. The findings hold importance in guiding couples' informed decision-making processes regarding their choice of genetic screening and diagnostic testing in case of intermediate risk for T21., (© 2024 The Author(s). Journal of Genetic Counseling published by Wiley Periodicals LLC on behalf of National Society of Genetic Counselors.)

Apitherapy has started to gain tremendous recognition because of extraordinary pharmacological importance of honeybee-related ingredients and their derivatives. There has been a renewed interest in the bee venom-based therapies. Interdisciplinary researchers are studying the chemistry and translational value of venom for effective cancer treatment. Bee venom and its major component, melittin, are cytotoxic in cancer cells. In this study, MTT and scratch assays were performed for analysis of melittin-mediated antimetastatic effects. QPCR was used for expression profiling of metastasis-related genes. Three anti-metastatic genes (BRMS1, DRG1, and KAI1/CD82) were studied for the first time after bee venom and melittin treatment in MDA-MB-231 breast cancer cells compared with normal breast cells, and two prometastatic genes (EGFR and WNT7B) were also examined. KAI1/CD82 and BRMS1 are the negative regulators of EGFR. WNT7B is a negative regulator of KAI1/CD82. Selective cytotoxicity of bee venom and melittin was found to be higher as compared to cisplatin. Melittin induced an increase in the expression of BRMS1 and DRG1, whereas bee venom upregulated DRG1 and KAI1/CD82 expression in breast cancer. WNT7B was downregulated in bee venom-treated breast cancer cells. Results suggested that bee venom/melittin exerted antimetastatic effects primarily through upregulation of BRMS1, DRG1, and KAI1/CD82, and downregulation of WNT7B., (© 2024 John Wiley & Sons Ltd.)

Competing Interests: The authors declare that they have no conflict of interest.

The COVID-19 pandemic has caused havoc around the globe since 2019 and is considered the largest global epidemic of the twentieth century. Although the first antiviral drug, Remdesivir, was initially introduced against COVID‑19, virtually no tangible therapeutic drugs exist to treat SARS-CoV-2 infection. FDA-approved Paxlovid (Nirmatrelvir supplemented by Ritonavir) was recently announced as a promising drug against the SARS-CoV-2 major protease (M pro ). Here we report for the first time the remarkable inhibitory potentials of lead epigenetic-targeting drugs (epi-drugs) against SARS-CoV-2 M pro . Epi-drugs are promising compounds to be used in combination with cancer chemotherapeutics to regulate gene expression. The search for all known epi-drugs for the specific inhibition of SARS-CoV-2 M pro was performed for the first time by consensus (three high-order program) molecular docking studies and end-state free energy calculations. Several epi-drugs were identified with highly comparable binding affinity to SARS-CoV-2 M pro compared to Nirmatrelvir. In particular, potent histone methyltransferase inhibitor EPZ005687 and DNA methyltransferase inhibitor Guadecitabine were prominent as the most promising epi-drug inhibitors for SARS-CoV-2 M pro . Long Molecular dynamics (MD) simulations (200 ns each) and corresponding MM-GBSA calculations confirmed the stability of the EPZ005687-M pro complex with MM-GBSA binding free energy (ΔG bind ) -48.2 kcal/mol (EPZ005687) compared to Nirmatrelvir (-44.7 kcal/mol). Taken together, the antiviral activities of the highlighted epi-drugs are reported beyond widespread use in combination with anti-cancer agents. The current findings therefore highlight as yet unexplored antiviral potential of epi-drugs suitable for use in patients struggling with chronic immunosuppressive disorders.Communicated by Ramaswamy H. Sarma.

Tragopogon coelesyriacus is a pharmacotherapeutic herbaceous plant belonging to the Asteraceae family and consumed as a vegetable. Here, the methanolic and water extracts of T. coelesyriacus were obtained from its aboveground parts (stem, leaves, and flowers), and the phytochemical potentials were investigated by LC-HRMS (liquid chromatography-high resolution mass spectrometry) analysis for the first time. The antibacterial, antifungal, and anticarcinogenic activities of T. coelesyriacus extracts were investigated using experimental and in silico methods. T. coelesyriacus methanol extract revealed remarkable inhibitory activity against Staphylococcus aureus , Pseudomonas aeruginosa , and Klebsiella pneumonia (MICs = 0.83, 1.67, and 1.67 mg/mL, respectively) compared to Escherichia coli and Enterobacter aerogenes (MIC = 53.3 mg/mL). Inhibitory effects of T. coelesyriacus methanolic extracts were also observed in all Candida species tested, with the highest inhibition on Candida krusei (MIC = 0.83 mg/mL), whereas no inhibitory effect was identified from the water extract. Additionally, both T. coelesyriacus methanolic (IC50 = 86 μg/mL) and water (IC50 = 92 μg/mL) extracts revealed significant selective anticarcinogenic effects on AR42J pancreatic cancer cells. HeLa and MDA-MB-231 cells were, however, more resilient to methanol and water extract, respectively. In silico analyses further elucidated the noteworthy antibacterial potential of keracyanin chloride on S. aureus MurB enzyme and the remarkable inhibitory potential of naringin on FYN kinase specific for pancreatic cancer (AR42J) development. In conclusion, T. coelesyriacus phytochemicals with antibacterial, antifungal, and anticancer properties were revealed for the first time, and molecular docking studies on potential targets confirmed good agreement with experimental findings. Therefore, the current studies on T. coelesyriacus provide the basis for investigating new pharmaceutical potentials of other Tragopogon members., (© 2024 The Author(s). Food Science & Nutrition published by Wiley Periodicals LLC.)

Competing Interests: The authors declare that they have no conflict of interest.

BACKGROUND AND OBJECTIVES: Children with tracheostomies are at increased risk of tracheostomy-related complications and require extra care. Standardized training programs for caregivers can improve tracheostomy care and reduce complications. In this study, we compared caregiver knowledge and skill scores after a standardized theoretical and practical training program on tracheostomy care (IStanbul PAediatric Tracheostomy (ISPAT) project) immediately and 1 year post-training and evaluated how this training affected the children's clinical outcomes., Materials and Methods: We included 32 caregivers (31 children) who had received standardized training a year ago and administered the same theoretical and practical tests 1 year after training completion. We recorded tracheostomy-related complications and the number and reasons for admission to the healthcare centers. All data just before the training and 1 year after training completion were compared., Results: After 1 year of training completion, the median number of correct answers on the theoretical test increased to 16.5 from 12 at pretest (p < 0.001). Compared with pretest, at 1-year post-training practical skills assessment scores, including cannula exchange and aspiration, were significantly higher (both p < 0.001) and mucus plug, bleeding, and stoma infection reduced significantly (p = 0.002, 0.022, and 0.004, respectively). Hands-on-training scores were better than pretest but declined slightly at 1 year compared to testing immediately after training. Emergency admission decreased from 64.5% to 32.3% (p = 0.013). Hospitalization decreased from 61.3% to 35.5% (p = 0.039)., Conclusion: Our findings indicate that caregiver training can lead to a persistent increase in knowledge and skill for as long as 1 year, as well as improvements in several measurable outcomes, although a slight decrease in scores warrants annual repetitions of the training program., (© 2023 Wiley Periodicals LLC.)

Purpose: Due to its link with the 2019 coronavirus, the multisystem inflammatory syndrome in children (MISC) has garnered considerable international interest. The aim of this study, in which MISC patients were evaluated multicenter, and the data of the third period of the Turk-MISC study group, to compare the clinical and laboratory characteristics and outcomes of MISC patients who did and did not require admission to an intensive care unit (ICU)., Methods: This retrospective multicenter observational study was carried out between June 11, 2021, and January 01, 2022. The demographics, complaints, laboratory results, system involvements, and outcomes of the patients were documented., Results: A total of 601 patients were enrolled; 157 patients (26.1%) required hospitalization in the intensive care unit (ICU). Median age was 8 years (interquartile range (IQR) 4.5-11.3 years. The proportion of Kawasaki disease-like features in the ICU group was significantly higher than in the non-ICU group (56.1% vs. 43.2% p = 0.006). The ICU group had considerably lower counts of both lymphocytes and platelets (lymphocyte count 900 vs. 1280 cells × μL, platelet count 153 vs. 212 cells × 10 3 / μL, all for p< 0.001). C-reactive protein, procalcitonin, and ferritin levels were significantly higher in the ICU group (CRP 164 vs. 129 mg/L, procalcitonin 9.2 vs. 2.2 μg/L, ferritin 644 vs. 334 μg/L, all for p< 0.001). Being between ages 5-12 and older than 12 increased the likelihood of hospitalization in the ICU by four [95% confidence intervals (CI)1.971-8.627] and six times (95% CI 2.575-14.654), respectively, compared to being between the ages 0-5. A one-unit increase in log D-dimer (µg/L) and log troponin (ng/L) was also demonstrated to increase the need for intensive care by 1.8 (95% CI 1.079-3.233) and 1.4 times (95% CI 1.133-1.789), respectively.     Conclusion: By comparing this study to our other studies, we found that the median age of MISC patients has been rising. Patients requiring an ICU stay had considerably higher levels of procalcitonin, CRP, and ferritin but significantly lower levels of lymphocyte and thrombocyte. In particular, high levels of procalcitonin in the serum might serve as a valuable laboratory marker for anticipating the need for intensive care., What Is Known: • Lymphopenia and thrombocytopenia were an independent predictor factors in patients with MISC who needed to stay in intensive care unit. • The possibility of the need to stay in the intensive care unit in patients with MISC who had Kawasaki disease-like findings was controversial compared with those who did not., What Is New: • A one-unit increase log D dimer and log troponin was demonstrated to require for intensive care unit by 1.8 and 1.4 times, respectively. • Serum procalcitonin levels had the best performance to predict stay in the intensive care unit stay., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Cancer has the second-highest mortality rate worldwide after cardiovascular disease. In addition, breast and cervical cancer are two of the leading causes of cancer-related deaths among women. The tumor microenvironment, which consists of fibroblasts, immune cells, cells that form blood vessels, and proteins, is a therapeutic target for cancer therapy. As part of the cellular microenvironment, glycosaminoglycan chondroitin sulfate is associated with various aspects of tumor progression and metastasis depending on the sulfate pattern of chondroitin sulfate. This study evaluated the roles of Microbial Chondroitin Sulfate (CS) and Commercial CS in tumor growth and metastasis comparatively using MDA-MB-231 metastatic breast cancer cells, HeLa cervical cancer cells, and normal fibroblasts. In addition, the role of CS types in wound healing was also assessed comparatively.  Microbial CS was more cytotoxic in MDA-MB-231 cells than HeLa compared to Commercial CS. Although both CS reduced cell viability in normal cells, the selective index of Microbial CS in MDA-MB-213 cells was higher than its commercial counterpart. In addition, the role of CS types in wound healing was also assessed comparatively. Both types of CS decreased the cell migration in MDA-MB-231 cancer cells, but HeLa cells were more sensitive to Microbial CS than Commercial CS to heal the wound. The wound healing of NIH3T3 cells after Microbial CS was similarly high to the healing after Commercial CS. This preliminary study shows that microbial CS produced by biotechnological methods from a recombinant source created by our team can be an effective therapeutic agent in various types of cancer.

Background: Cystic fibrosis (CF) is an autosomal recessive disorder caused by CF transmembrane conductance regulator (CFTR) genetic variants. CFTR modulators improve pulmonary function and reduce respiratory infections in CF. This study investigated the clinical and laboratory follow-up parameters over 1 year in patients with CF who could not receive this treatment., Methods: This retrospective cohort study included 2018 and 2019 CF patient data from the CF registry of Turkey. Demographic and clinical characteristics of 294 patients were assessed, who had modulator treatment indications in 2018 but could not reach the treatment., Results: In 2019, patients younger than 18 years had significantly lower BMI z-scores than in 2018. During the 1-year follow-up, forced expiratory volumes (FEV1) and FEV1 z-scores a trend toward a decrease. In 2019, chronic Staphylococcus aureus colonization, inhaled antipseudomonal antibiotic use for more than 3 months, oral nutritional supplement requirements, and oxygen support need increased., Conclusions: Patients who had indications for modulator treatments but were unable to obtain them worsened even after a year of follow-up. This study emphasized the importance of using modulator treatments for patients with CF in our country, as well as in many countries worldwide., (© 2023 Wiley Periodicals LLC.)

Hardaliye, as one of the oldest and lesser known traditional beverages, is produced using red grape pomace from wine production. This drink production is achieved through lactic acid fermentation, with the addition of sour cherry leaves and mustard seeds-either heat-treated, grinded, or whole-in various concentrations. Hardaliye has a very short shelf life; thus, efforts have recently been made to process hardaliye with novel processing technologies in order to achieve shelf-life extension. Therefore, the high-hydrostatic-pressure (HHP) processing of hardaliye was performed to determine its impact on important properties, including in microbial inactivation and shelf-life extension, with respect to a Box-Behnken experimental design. Maximum log reductions of 5.38 ± 0.6, 5.10 ± 0.0, 5.05 ± 0.2, and 4.21 ± 0.0 with HHP were obtained for Brettanomyces bruxellensis , total mesophilic aerobic bacteria, Lactobacillus brevis , and total mold and yeast, respectively. The processing parameters of 490 MPa and 29 °C for 15 min were found as the optimal conditions, with the response variables of an optical density at 520 nm and the inactivation of L. brevis . The samples processed at the optimal conditions were stored at both 4 and 22 °C for 228 d. While the non-treated control samples at 4 and 22 °C were spoiled at 15 and 3 d, the HHP-treated samples were spoiled after 228 and 108 d at 4 and 22 °C, respectively.

Bee venom is a natural mixture and candidate anti-cancer agent with selective cytotoxic effect on some cancer cells. However, the cellular mechanisms of how bee venom selectively targets cancer cells remain elusive. The aim of this study was to reveal the genotoxic effect of bee venom in concordance with the location of β-actin protein throughout the nucleus or/and cytoplasm. For this aim, the level of H2AX phosphorylation (γH2AX) and intracellular location of β-actin were assessed by immunofluorescence in liver (HEPG2) and metastatic breast (MDA-MB-231) cancer cell lines compared to normal fibroblasts (NIH3T3) after bee venom treatment. Colocalisation profiles of γH2AX and β-actin in each cell line were also analysed. The results showed that the levels of γH2AX staining decreased in normal cells but increased in cancer cells. The majority of β-actin was localised within the cytoplasm of normal cells after bee venom treatment, but it was mostly accumulated within the nucleus in cancer cells. Colocalisation of β-actin and γH2AX both in nucleus and cytoplasm was induced in each cancer cell by different patterns. The results showed that normal and cancerous cells had different responses against bee venom, and suggested that bee venom induced a cellular response by the interaction between γH2AX and β-actin., (© 2023 Wiley-VHCA AG, Zurich, Switzerland.)

Background: We aimed to determine the number of cystic fibrosis (CF) patients recorded in the Cystic Fibrosis Registry of Türkiye (CFRT) who were in need of lung transplantation (LT) referral and examine clinical differences between patients who were LT candidates due to rapid forced expiratory volume in one second (FEV₁) decline and LT candidates without rapid FEV₁ decline in the last year to identify a preventable cause in patients with such rapid FEV₁ decline., Methods: All CF patients recorded in the CFRT in 2018 were evaluated in terms of LT. Patients were divided into those with FEV₁ below 50% and in need of LT due to a decrease of 20% or more in the previous year (Group 1) and those who did not have FEV₁ decline of more than 20% in the previous year but had other indications for LT (Group 2). Demographic and clinical features were compared between the two groups., Results: Of 1488 patients registered in CFRT, 58 had a need for LT. Twenty patients were included in Group 1 and others in Group 2. Our findings did not reveal any significant variations in treatment, chronic infection status, or complications between the two groups. The average weight z-score was significantly higher in Group 1. Positive correlations were detected between weight z-score and FEV₁ in 2017 in Group 1 and between FEV₁ values in 2017 and 2018 in Group 2., Conclusions: There appears to be a relationship between the nutritional status and weight z-scores of CF patients and pulmonary function, which may indirectly affect the need for lung transplantation referral.

Recently, AZ31B magnesium alloy has been widely employed in automotive, aerospace, and bio implant industries due to its light-weight and biocompatibility properties. However, the equilibrium of ductility and strength of this material and the negativity brought by its poor wear behavior have limited its versatile use. Friction stir processing (FSP) has been commonly used as severe plastic deformation method for improving mechanical and tribological properties of metal sheets. The effect of this method on the biocompatibility of materials is a matter of curiosity that should be emphasized. So, the present study aims to investigate the effect of friction stir process on the mechanical, tribological, and biocompatibility properties of AZ31B magnesium alloy. It is observed that FSP enhanced the tensile properties of the alloy but decreased its elongation. It was determined that the base material exhibited ductile character on the fracture surface of the specimens, and mixed ductile/brittle fracture was evident with the FSP. In the FSP zone, the hardness value was improved by 17% compared to the base material. Also, the wear performance of the alloy enhanced in ambient air and Simulated Body Fluid (SBF) solution. Wear properties in SBF solution were better due to less adhesive bonds between the friction surfaces. This assessment was supported by SEM images of the wear path and surface of counter bodies. On the other hand, FSPed AZ31B alloy materials with improved strength properties were not cytotoxic for human gingival fibroblasts, and these results may suggest that the materials are safe for clinical uses.

Presence of aflatoxins in agricultural products is a worldwide problem. Because of their high heat stability and resistance to most of the food processing technologies, aflatoxin degradation is still a big challenge. Thus, efficacy of pulsed electric fields (PEF) by energies ranging from 0.97 to 17.28 J was tested to determine changes in quality properties in red pepper flakes, mitigation of aflatoxins, inactivation of aflatoxin producing Aspergillus parasiticus, reduction in aflatoxin mutagenity, and modelling of A. parasiticus inactivation in addition to aflatoxin mitigation. Maximum inactivation rate of 64.37 % with 17.28 J was encountered on the mean initial A. parasiticus count. A 99.88, 99.47, 97.75, and 99.58 % reductions were obtained on the mean initial AfG 1 , AfG 2 , AfB 1 , and AfB 2 concentrations. PEF treated samples by 0.97, 1.36, 5.76, and 17.28 J at 1 μg/plate, 0.97, 1.92, 7.78, 10.80 J at 10 μg/plate, and 0.97, 1.92, 2.92, 4.08, 5.76, 4.86, 6.80, 9.60, 10.80, and 10.89 J at 100 μg/plate were not mutagenic. Modelling with gradient boosting regression tree (GBRT), random forest regression (RFR), and artificial neural network (ANN) provided the lowest RMSE and highest R 2 value for GBRT model for the predicted inactivation of A. parasiticus, whereas ANN model provided the lowest RMSE and highest R 2 for predicted mitigation of AfG 1 , AfB 1 , and AfB 2 . PEF treatment possess a viable alternative for aflatoxin degradation with reduced mutagenity and without adverse effect on quality properties of red pepper flakes., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Functional recovery is provided by some neurotrophic factors released from the near vicinity of the injury site. Ultrasound treatment is known to increase neurotrophic factor expression. This study was aimed at determining the effect of ultrasound treatment on the expression of vascular endothelial growth factor (VEGF), its receptors and new vessel formation after facial nerve injury. Sixty-four Wistar rats were divided into four groups: control (group 1), sham (group 2), facial-facial coaptation (group 3), and facial-facial coaptation and ultrasound treatment (group 4). Animals in each group were evaluated on the 14th and 28th days. Immunohistochemical staining and electrophysiological and gene-level evaluations were performed for the expression of VEGF and its receptors. When the results were evaluated, it was determined that VEGF, VEGFR1 (VEGF receptor 1), VEGFR2 (VEGF receptor 2) and CD31 levels were significantly higher in groups 3 and 4 compared with the control and sham groups. The increase in these values was more prominent after 28 d of ultrasound treatment than all groups. Electrophysiological results revealed similar evident functional improvement in group 4 with decreased latency and increased amplitudes compared with group 3. Our findings suggest that ultrasound treatment might promote injured facial nerve regeneration by stimulating release of VEGF and its receptors and may result in functional improvement., Competing Interests: Declaration of Competing Interest The authors have no conflicts of interest to declare., (Copyright © 2022 World Federation for Ultrasound in Medicine & Biology. Published by Elsevier Inc. All rights reserved.)

Background: Accumulation of unfolded or misfolded proteins in the cellular environment result in ER stress and activates the unfolded protein response (UPR). The UPR alleviates ER stress and restores homeostasis, but it triggers cell death under prolonged stress. Here, we aimed to investigate the involvement of Sec71, an Arf-GEF involved in vesicular transport, in the tunicamycin-induced ER stress response. Since deubiquitinases and ER stress are known to be closely linked, we investigated this response by evaluating the potential role of Ubp2, a deubiquitinase, in the ER stress response in fission yeast., Methods and Results: Tunicamycin-induced ER stress responses were assessed by analyzing cell viability, apoptosis, intracellular oxidation levels, and proteasomal activities in sec71 and ubp2-deficient cells. The cell viability of Δsec71 and Δubp2 decreased after exposure to 0.5 µg/mL tunicamycin. Deleting either ubp2 or sec71 genes significantly decreased proteasomal activity and sensitized cells to ER stress, resulting in increased apoptosis compared with wild-type cells after tunicamycin treatment. DCFDA (2,7-dichlorodihydrofluorescein diacetate) reduction increased in correlation with apoptosis observed in the mutant cells, indicating higher levels of reactive oxygen species., Conclusions: The results highlight the involvement of S. pombe Ubp2 in the known role of the ubiquitin-proteasome system in the ER stress response. We hypothesise that Sec71 is associated with ER homeostasis, and our findings on Sec71 provide new insight into the regulation of cell death mechanisms arising from the ER stress., (© 2022. The Author(s), under exclusive licence to Springer Nature B.V.)

Introduction: The diagnosis of childhood tuberculosis is difficult and most of the patients are diagnosed clinically. The objective of this study is to reveal the diagnostic and therapeutic components of childhood pulmonary tuberculosis and to analyze the changes that occurred in our country over the years., Methodology: All patients diagnosed with tuberculosis between 2006 and 2016 were included. Demographic characteristics, diagnostic and treatment outcomes were recorded and patients were followed up prospectively., Results: A total of 492 patients were included in the study. 97% had Bacillus Calmette-Guerin vaccine, 36% were diagnosed with microbiologically-confirmed tuberculosis and 64% were diagnosed with clinically-proven tuberculosis. 94% of the patients had symptoms consistent with tuberculosis, all patients had radiologic findings, 74% had a history of tuberculosis contact and 63% had tuberculin skin test positivity. The diagnoses included primary tuberculosis in 62%, secondary tuberculosis in 21%, progressive primary tuberculosis in 13% and miliary tuberculosis in 4%. 48% of the patients received a treatment regimen containing three drugs as the initial treatment, and drug-related side effects developed in 12%. Isoniazid resistance was detected in 13% of the patients and rifampicin resistance was detected in 8%. None of the patients died due to tuberculosis. In the last 50 years in Turkey, the rates of Bacillus Calmette-Guerin vaccination and diagnosis of tuberculosis cases have increased and the mortality rates have decreased over the years., Conclusions: Our study is one of the few prospective studies and revealed the differences between the recent data and the past 50 years in childhood tuberculosis in Turkey., Competing Interests: No Conflict of Interest is declared, (Copyright (c) 2022 Aysegul Dogan Demir, Arif Kut, Erhan Ozaydin, Mustafa Nursoy, Fatma Betul Cakir, Feyza Ustabas Kahraman, Ufuk Erenberk, Selcuk Uzuner, Abdulhamit Collak, Zeynep Ebru Cakin, Erkan Cakir.)

Background: Tracheostomy-related morbidity and mortality mainly occur due to decannulation, misplacement, or obstruction of the tube. A standardized training can improve the skills and confidence of the caregivers in tracheostomy care (TC)., Objective: Our primary aim was to evaluate the efficiency of standardized training program on the knowledge and skills (changing-suctioning the tracheostomy tube) of the participants regarding TC., Materials and Methods: Sixty-five caregivers of children with tracheostomy were included. First, participants were evaluated with written test about TC and participated in the practical tests. Then, they were asked to participate in a standardized training session, including theoretical and practical parts. Baseline and postintervention assessments were compared through written and practical tests conducted on the same day., Results: A significant improvement was observed in the written test score after the training. The median number of correct answers of the written test including 23 questions increased 26%, from 12 to 18 (p < .001). The median number of correct steps in tracheostomy tube change (from 9 to 16 correct steps out of 16 steps, 44% increase) and suctioning the tracheostomy tube (from 9 to 17 correct steps out of 18 steps, 44% increase) also improved significantly after the training (p < .001, for both)., Conclusion: Theoretical courses and practical hands-on-training (HOT) courses are highly effective in improving the practices in TC. A standardized training program including HOT should be implemented before discharge from the hospital. Still there is a need to assess the impact of the program on tracheostomy-related complications, morbidity, and mortality in the long term., (© 2021 Wiley Periodicals LLC.)

Objective: Having a child with a chronic illness is a source of stress for the whole family, especially the primary caregiver. The aim of this study was to evaluate the associations between caregiver burden and both the caregiver's and child's psychological symptoms in a cohort of children with systemic lupus erythematosus (SLE)., Methods: Thirty-four patients (aged 9-18 years) with childhood-onset SLE and their caregivers participated in this study. The control group was composed of healthy children and their caregivers. Questionnaires were used to evaluate caregiver burden and the psychological status of parents and children and adolescents with and without SLE., Results: No significant difference was found between the study and control groups for caregiver burden, anxiety and depression in parents, and psychological status in children. Caregiver burden was positively correlated with parent's depression, anxiety, and behavioral and peer problems of the children, and it was negatively correlated with the children's prosocial behaviors. According to regression analyses, the parents' depression and children's peer relationship had a positive effect on caregiver burden scores., Conclusion: Physicians should be aware of the presence of psychological symptoms in patients with childhood-onset SLE and their caregivers because it can affect caregiver burden and the caregiver's psychological state. Key points •Caregiver burden was positively correlated with parent's depression and anxiety. •Caregiver burden was positively correlated with children's behavioral and peer problems. •Caregiver burden was negatively correlated with child's prosocial behaviors., (© 2021. International League of Associations for Rheumatology (ILAR).)

Background and Objective: This study aims to evaluate clinical and radiological findings and treatment outcomes of the patients with PIBO., Methods: One hundred fourteen children were enrolled. Initial demographic and clinical findings were evaluated. Pre- and post-treatment clinical and radiological findings were compared., Results: The median age of the patients at initial pulmonary injury was 7,2 months, the median age at diagnosis was 17.5 months. Persistent wheezing was the most common complaint. Thirty-five patients had mechanical ventilation history. 82,5% of patients had clinical improvement. Bronchiectasis, atelectasis, hyperinflation and air trapping in HRCT improved significantly with treatment. Post-treatment Bhalla scores decreased from 8.3 to 6.5 (p= 0,001). Improvement was observed in radiological and clinical findings after treatment., Conclusions: This study is one of the largest studies in the literature and one of the few studies that evaluate clinical and radiological outcomes of patients with PIBO., Competing Interests: Declaration of Competing Interest No conflict of interest was declared by the authors., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Immunostaining is one of the advantageous methods for the qualitative analysis of cellular markers of interest. Immuno-stained cells are typically analyzed by fluorescence microscopy or flow cytometry. Flow cytometry has the advantage of being able to process large numbers of cells in a short time thus enhancing its quantitative capacity. The staining protocol typically includes fixation of cells followed by permeabilization, blocking procedures to reduce non-specific binding of the label, and staining with specific antibodies labeled directly or indirectly with fluorescence-tags. Important controls include staining with a relevant non-immune antibody to identify any non-specific imminent globally binding and measurements in the absence of any fluorescent tag to detect non-specific sources of fluorescence. The most common source of non-specific fluorescence is caused by autofluorescence of naturally occurring chemicals within the cells of interest. In this study, we found high levels of cellular autofluorescence in mouse embryonic fibroblasts, at levels that interfered with the detection of a number of cellular antigens using common fluorophores. This autofluorescence was detected in three of the four fluorescence channels restricting useful analysis to only one channel (red) on the instrument. The study highlights an important limitation to immunostaining techniques and reinforces the need for the use of a thorough set of controls to ensure specificity of quantitative analysis.

Brucellosis is one of the most common zoonosis worldwide. It is still endemic in many regions of the world. A 6-year-old female was admitted to the emergency department (ED) due to a sudden change in consciousness, urinary incontinence, vomiting, and difficulty in walking. Neurological examination demonstrated abducens nerve paralysis, mild-to-moderate motor deficit in hemiparesis in the left arm. Brain magnetic resonance imaging showed a hemorrhagic focus at the right frontal lobe and thrombosis in the superior sagittal sinus of the brain. The diagnosis of neurobrucellosis was confirmed by identifying Brucella spp. in the blood culture on the day 6 of pediatric intensive care unit admission; thus, trimethoprim-sulfamethoxazole and rifampicin, and ceftriaxone were promptly initiated. Despite neuroprotective management and acetazolamide, the patient's neurological problems and high intracranial pressure (ICP) persisted. An external ventricular drainage tube and a Codman ICP monitor were placed to be on the consent vigilance of the patient's neurological condition. The patient's ICP continued to increase despite the current treatment regimen; therefore, a decompressive bitemporal craniectomy was performed. The ICP level of the patient returned to its normal range immediately after the craniectomy. The patient did not have any notable neurologic sequelae at the first-year follow-up. Neurobrucellosis is a rare complication of systemic brucellosis and may present as meningitis, encephalitis, myelitis, radiculitis, and/or neuritis. Herein, we describe a six-year-old girl with brucellosis complicated with cerebral vein thrombosis. This case illustrates the need for close monitoring of patients with unexplained neurological signs or symptoms for brucellosis in endemic areas., Competing Interests: None declared., (Thieme. All rights reserved.)

Immunostaining (also called as immunofluorescence) is a fluorescence labeling method to stain one or more epitopes of interest on DNA and/or protein using specific antibodies. Cytosine modifications can be detected quantitatively by immunostaining. The protocol commonly includes sequential steps. These include fixation, permeabilization, antigen retrieval, blocking, incubation with primary and secondary antibodies, and visualization under the microscope followed by image-based intensity analysis of staining. Each step is important, but antigen retrieval is especially necessary for DNA epitopes such as cytosine modifications as antibodies can access cytosines in DNA only once the DNA double-strand is denatured and DNA-packaging proteins have been removed. Hydrochloric acid is commonly used for this purpose. However, there are additional treatments with enzymes to enhance antigen retrieval and improve the detection by increasing staining intensity. This chapter describes current methodology for improving antigen retrieval for the staining of the cytosine modifications 5'-methylcytosine (5meC), 5'-hydroxymethylcytosine (5hmC), 5'-formylcytosine (5fC), and 5'-carboxycytosine (5caC).

The menisci play a vital role in the mechanical function of knee joint. Unfortunately, meniscal tears often occur. Meniscectomy is a surgical treatment for meniscal tears; however, mechanical changes in the knee joint after meniscectomy is a risk factor to osteoarthritis (OA). The objective of this study was to investigate the altered cartilage mechanics of different medial meniscectomies using a poromechanical model of the knee joint. The cartilaginous tissues were modeled as nonlinear fibril-reinforced porous materials with full saturation. The ligaments were considered as anisotropic hyperelastic and reinforced by a fibrillar collagen network. A compressive creep load of ¾ body weight was applied in full extension of the right knee during 200 s standing. Four finite element models were developed to simulate different meniscectomies of the joint using the intact model as the reference for comparison. The modeling results showed a higher load support in the lateral than medial compartment in the intact joint, and the difference in the load share between the compartments was augmented with medial meniscectomy. Similarly, the contact and fluid pressures were higher in the lateral compartment. On the other hand, the medial meniscus in the normal joint experienced more loading than the lateral one. Furthermore, the contact pressure distribution changed with creep, resulting in a load transfer between cartilage and meniscus within each compartment while the total load born by the compartment remained unchanged. This study has quantified the altered contact mechanics on the type and size of meniscectomies, which may be used to understand meniscal tear or support surgical decisions., (Copyright © 2020 by ASME.)