Brain cholesterol and Alzheimer's disease: challenges and opportunities in probe and drug development.

Cholesterol homeostasis is impaired in Alzheimer's disease; however, attempts to modulate brain cholesterol biology have not translated into tangible clinical benefits for patients to date. Several recent milestone developments have substantially improved our understanding of how excess neuronal cholesterol contributes to the pathophysiology of Alzheimer's disease. Indeed, neuronal cholesterol was linked to the formation of amyloid-ß and neurofibrillary tangles through molecular pathways that were recently delineated in mechanistic studies. Furthermore, remarkable advances in translational molecular imaging have now made it possible to probe cholesterol metabolism in the living human brain with PET, which is an important prerequisite for future clinical trials that target the brain cholesterol machinery in Alzheimer's disease patients-with the ultimate aim being to develop disease-modifying treatments. This work summarizes current concepts of how the biosynthesis, transport and clearance of brain cholesterol are affected in Alzheimer's disease. Further, current strategies to reverse these alterations by pharmacotherapy are critically discussed in the wake of emerging translational research tools that support the assessment of brain cholesterol biology not only in animal models but also in patients with Alzheimer's disease.

miR-221 and Parkinson's disease: A biomarker with therapeutic potential.

Parkinson's disease (PD) is a neurodegenerative disorder characterized by the loss of dopaminergic neurons in the substantia nigra, leading to various motor and non-motor symptoms. Several cellular and molecular mechanisms such as alpha-synuclein (α-syn) accumulation, mitochondrial dysfunction, oxidative stress and neuroinflammation are involved in the pathogenesis of this disease. MicroRNAs (miRNAs) play important roles in post-transcriptional gene regulation. They are typically about 21-25 nucleotides in length and are involved in the regulation of gene expression by binding to the messenger RNA (mRNA) molecules. miRNAs like miR-221 play important roles in various biological processes, including development, cell proliferation, differentiation and apoptosis. miR-221 promotes neuronal survival against oxidative stress and neurite outgrowth and neuronal differentiation. Additionally, the role of miR-221 in PD has been investigated in several studies. According to the results of these studies, (1) miR-221 protects PC12 cells against oxidative stress induced by 6-hydroxydopamine; (2) miR-221 prevents Bax/caspase-3 signalling activation by stopping Bim; (3) miR-221 has moderate predictive power for PD; (4) miR-221 directly targets PTEN, and PTEN over-expression eliminates the protective action of miR-221 on p-AKT expression in PC12 cells; and (5) miRNA-221 controls cell viability and apoptosis by manipulating the Akt signalling pathway in PD. This review study suggested that miR-221 has the potential to be used as a clinical biomarker for PD diagnosis and stage assignment.

Genetic association of ACE2 and TMPRSS2 polymorphisms with COVID-19 severity; a single centre study from Egypt.

BACKGROUND: Since the emergence of the COVID-19 infection in China, it has caused considerable morbidity, mortality, and economic burden. It causes the vast majority of clinical manifestations, ranging from mild or even no symptoms to severe respiratory failure. There are many risk factors for severe COVID-19, such as old age, male gender, and associated comorbidities. A major role for genetic factors may exist. The SARS-CoV-2 virus enters the cell primarily through ACE2 receptors. rs2285666 is one of many polymorphisms found in the ACE2 receptor gene. To enable endosome-independent entry into target cells, the transmembrane protease serine-type 2 (TMPRSS2) is necessary to cleave the virus' spike (S) glycoprotein. TMPRSS2 is characterized by an androgen receptor element. The rs12329760 polymorphism in TMPRSS2 may explain different genetic susceptibilities to COVID-19. METHOD: This cross-sectional study was held in Mansoura University Hospitals during the period from June 2020 to April 2022 on patients who had mild and severe COVID-19. Demographic, clinical, and laboratory data were collected, and the TaqMan real-time polymerase chain was used for allelic discrimination in the genotyping of rs2285666 and rs12329760. RESULTS: This study included 317 Egyptian patients, aged from 0.2 to 87 years. Males were 146, while females were 171. They were divided into mild and severe groups (91 and 226 patients, respectively) based on their clinical symptoms. There was a significant association between COVID-19 severity and male gender, hypertension, diabetes mellitus, and high CRP. The genotype and allele frequency distributions of the ACE2 rs2285666 polymorphism showed no significant association with the severity of COVID-19 in both. In contrast, in TMPRSS2 rs12329760 minor T allele and CT, TT genotypes were significantly associated with a reduced likelihood of developing severe COVID-19. CONCLUSION: Our study indicates that the ACE2 rs2285666 polymorphism is not related to the severity of COVID-19, whether genotypes or alleles. In TMPRSS2 rs12329760, the dominant model and T allele showed significantly lower frequencies in severe cases, with a protective effect against severity. The discrepancies with previous results may be due to variations in other ACE2 receptor-related genes, inflammatory mediators, and coagulation indicators. Haplotype blocks and differences in racial makeup must be taken into consideration. Future research should be done to clarify how ethnicity affects these polymorphisms and how other comorbidities combine to have an additive effect.

Insight into hazards and control of transfusion-transmitted infections in Egypt: A narrative review.

Blood transfusion is a critical life-saving medical intervention, but it carries the risk of transfusion-transmitted infections (TTIs) that can lead to serious consequences. TTIs include viral, bacterial, parasitic, and prion infections, transmitted through asymptomatic donor blood, contamination of stored blood products, or transfusion-related immunosuppression. Recognized global agents posing challenges to blood safety include human immunodeficiency virus (HIV), hepatitis C virus (HCV), hepatitis B virus (HBV), Syphilis, etc. Emerging pathogens like SARS-CoV-2, hepatitis E, and others present additional risks. The residual risk of TTIs, representing the likelihood of infected donations passing screening tests, varies globally. High-income countries generally show lower prevalence rates than low-income countries. In Egypt, the estimated prevalence rates for HIV, HBV, HCV, and syphilis markers among the donors are 0.23 %, 0.76 %, 2.33 %, and 0.24 %, respectively. In Egypt, specific residual risk estimates are scarce, but prevalence rates for key infections highlight existing challenges. The World Health Organization promotes a global blood safety strategy, advocating for national blood systems, voluntary non-remunerated donors, and quality-assured testing. Despite these measures, the establishment of a haemovigilance system which is critical for monitoring and preventing adverse events, including TTIs, is reported as lacking in Egypt. This highlights the importance of comprehensive surveillance and safety measures in the blood donation process to ensure universal access to safe blood. Primary health care can play a pivotal role in preventing TTIs.

Rest/stress myocardial perfusion imaging by positron emission tomography with 18F-Flurpiridaz: A feasibility study in mice.

BACKGROUND: Myocardial perfusion imaging by positron emission tomography (PET-MPI) is the current gold standard for quantification of myocardial blood flow. 18F-flurpiridaz was recently introduced as a valid alternative to currently used PET-MPI probes. Nonetheless, optimum scan duration and time interval for image analysis are currently unknown. Further, it is unclear whether rest/stress PET-MPI with 18F-flurpiridaz is feasible in mice. METHODS: Rest/stress PET-MPI was performed with 18F-flurpiridaz (0.6-3.0 MBq) in 27 mice aged 7-8 months. Regadenoson (0.1 µg/g) was used for induction of vasodilator stress. Kinetic modeling was performed using a metabolite-corrected arterial input function. Image-derived myocardial 18F-flurpiridaz uptake was assessed for different time intervals by placing a volume of interest in the left ventricular myocardium. RESULTS: Tracer kinetics were best described by a two-tissue compartment model. K1 ranged from 6.7 to 20.0 mL·cm-3·min-1, while myocardial volumes of distribution (VT) were between 34.6 and 83.6 mL·cm-3. Of note, myocardial 18F-flurpiridaz uptake (%ID/g) was significantly correlated with K1 at rest and following pharmacological vasodilation for all time intervals assessed. However, while Spearman's coefficients (rs) ranged between 0.478 and 0.681, R2 values were generally low. In contrast, an excellent correlation of myocardial 18F-flurpiridaz uptake with VT was obtained, particularly when employing the averaged myocardial uptake from 20 to 40 min post tracer injection (R2 ≥ 0.98). Notably, K1 and VT were similarly sensitive to pharmacological vasodilation induction. Further, mean stress-to-rest ratios of K1, VT, and %ID/g 18F-flurpiridaz were virtually identical, suggesting that %ID/g 18F-flurpiridaz can be used to estimate coronary flow reserve (CFR) in mice. CONCLUSION: Our findings suggest that a simplified assessment of relative myocardial perfusion and CFR, based on image-derived tracer uptake, is feasible with 18F-flurpiridaz in mice, enabling high-throughput mechanistic CFR studies in rodents.

Does sequential examination under anaesthesia provide a reliable method to determine a management plan for unstable lateral compression pelvic ring injuries? a prospective study.

PURPOSE: To assess the reliability of sequential examination under anaesthesia (EUA) to determine pelvic instability and to evaluate radiological and functional outcomes in unstable lateral compression (LC) injuries. METHODS: A prospective case series study was conducted from 2020 to 2022 at a university hospital on 43 cases with LC injuries that met the inclusion criteria. Sequential EUA was carried out in three steps. Posterior-only fixation or anterior-posterior fixation was done according to the algorithm. Each patient was followed up for at least 12 months, both radiologically and functionally. RESULTS: Forty cases proved unstable and were fixed. None showed secondary displacement in the anterior-posterior fixation group. However, five cases (19.2%) of the posterior-only fixation group showed secondary displacement with a mean of 5.9 mm. Four cases of them had tetra-ramic injuries. There is a high tendency for secondary displacement at 14.5 mm or more preoperative displacement of the rami. Patients with secondary displacement showed comparable functional outcome scores to patients without secondary displacement. Posterior-only fixation showed shorter operative time, lesser radiological exposure, blood loss and iatrogenic nerve injury than anterior-posterior fixation. CONCLUSION: EUA is a reliable method to determine pelvic instability and management plan for LC fractures with unilateral anterior ring injury. Anterior-posterior fixation is needed if there is a tetra-ramic fracture or initial anterior ring displacement of 14.5 mm or more, irrespective of EUA.

Characterization in nonhuman primates of (R)-[18F]OF-Me-NB1 and (S)-[18F]OF-Me-NB1 for imaging the GluN2B subunits of the NMDA receptor.

PURPOSE: GluN2B containing N-methyl-D-aspartate receptors (NMDARs) play an essential role in neurotransmission and are a potential treatment target for multiple neurological and neurodegenerative diseases, including stroke, Alzheimer's disease, and Parkinson's disease. (R)-[18F]OF-Me-NB1 was reported to be more specific and selective than (S)-[18F]OF-Me-NB1 for the GluN2B subunits of the NMDAR based on their binding affinity to GluN2B and sigma-1 receptors. Here we report a comprehensive evaluation of (R)-[18F]OF-Me-NB1 and (S)-[18F]OF-Me-NB1 in nonhuman primates. METHODS: The radiosynthesis of (R)-[18F]OF-Me-NB1 and (S)-[18F]OF-Me-NB1 started from 18F-fluorination of the boronic ester precursor, followed by removal of the acetyl protecting group. PET scans in two rhesus monkeys were conducted on the Focus 220 scanner. Blocking studies were performed after treatment of the animals with the GluN2B antagonist Co101,244 or the sigma-1 receptor antagonist FTC-146. One-tissue compartment (1TC) model and multilinear analysis-1 (MA1) method with arterial input function were used to obtain the regional volume of distribution (VT, mL/cm3). Occupancy values by the two blockers were obtained by the Lassen plot. Regional non-displaceable binding potential (BPND) was calculated from the corresponding baseline VT and the VND derived from the occupancy plot of the Co101,244 blocking scans. RESULTS: (R)- and (S)-[18F]OF-Me-NB1 were produced in > 99% radiochemical and enantiomeric purity, with molar activity of 224.22 ± 161.69 MBq/nmol at the end of synthesis (n = 10). Metabolism was moderate, with ~ 30% parent compound remaining for (R)-[18F]OF-Me-NB1 and 20% for (S)-[18F]OF-Me-NB1 at 30 min postinjection. Plasma free fraction was 1-2%. In brain regions, both (R)- and (S)-[18F]OF-Me-NB1 displayed fast uptake with slower clearance for the (R)- than (S)-enantiomer. For (R)-[18F]OF-Me-NB1, both the 1TC model and MA1 method gave reliable estimates of regional VT values, with MA1 VT (mL/cm3) values ranging from 8.9 in the cerebellum to 12.8 in the cingulate cortex. Blocking with 0.25 mg/kg of Co101,244 greatly reduced the uptake of (R)-[18F]OF-Me-NB1 across all brain regions, resulting in occupancy of 77% and VND of 6.36, while 0.027 mg/kg of FTC-146 reduced specific binding by 30%. Regional BPND, as a measure of specific binding signals, ranged from 0.40 in the cerebellum to 1.01 in the cingulate cortex. CONCLUSIONS: In rhesus monkeys, (R)-[18F]OF-Me-NB1 exhibited fast kinetics and heterogeneous uptake across brain regions, while the (S)-enantiomer displayed a narrower dynamic range of uptake across regions. A Blocking study with a GluN2B antagonist indicated binding specificity. The value of BPND was > 0.5 in most brain regions, suggesting good in vivo specific binding signals. Taken together, results from the current study demonstrated the potential of (R)-[18F]OF-Me-NB1 as a useful radiotracer for imaging the GluN2B receptors.

Role of sex hormones in modulating myocardial perfusion and coronary flow reserve.

BACKGROUND: A growing body of evidence highlights sex differences in the diagnostic accuracy of cardiovascular imaging modalities. Nonetheless, the role of sex hormones in modulating myocardial perfusion and coronary flow reserve (CFR) is currently unclear. The aim of our study was to assess the impact of female and male sex hormones on myocardial perfusion and CFR. METHODS: Rest and stress myocardial perfusion imaging (MPI) was conducted by small animal positron emission tomography (PET) with [18F]flurpiridaz in a total of 56 mice (7-8 months old) including gonadectomized (Gx) and sham-operated males and females, respectively. Myocardial [18F]flurpiridaz uptake (% injected dose per mL, % ID/mL) was used as a surrogate for myocardial perfusion at rest and following intravenous regadenoson injection, as previously reported. Apparent coronary flow reserve (CFRApp) was calculated as the ratio of stress and rest myocardial perfusion. Left ventricular (LV) morphology and function were assessed by cardiac magnetic resonance (CMR) imaging. RESULTS: Orchiectomy resulted in a significant decrease of resting myocardial perfusion (Gx vs. sham, 19.4 ± 1.0 vs. 22.2 ± 0.7 % ID/mL, p = 0.034), while myocardial perfusion at stress remained unchanged (Gx vs. sham, 27.5 ± 1.2 vs. 27.3 ± 1.2 % ID/mL, p = 0.896). Accordingly, CFRApp was substantially higher in orchiectomized males (Gx vs. sham, 1.43 ± 0.04 vs. 1.23 ± 0.05, p = 0.004), and low serum testosterone levels were linked to a blunted resting myocardial perfusion (r = 0.438, p = 0.020) as well as an enhanced CFRApp (r = -0.500, p = 0.007). In contrast, oophorectomy did not affect myocardial perfusion in females. Of note, orchiectomized males showed a reduced LV mass, stroke volume, and left ventricular ejection fraction (LVEF) on CMR, while no such effects were observed in oophorectomized females. CONCLUSION: Our experimental data in mice indicate that sex differences in myocardial perfusion are primarily driven by testosterone. Given the diagnostic importance of PET-MPI in clinical routine, further studies are warranted to determine whether testosterone levels affect the interpretation of myocardial perfusion findings in patients.

Serum MicroRNAs as predictors for fibrosis progression and response to direct-acting antivirals treatment in hepatitis C virus genotype-4 Egyptian patients.

BACKGROUND: MicroRNA (miRNAs) are small non-coding molecules that play an important role in hepatitis C virus (HCV) replication and liver diseases progression. The current study aimed to evaluate serum miRNAs as potential biomarkers for diagnosis, monitoring of fibrosis progression and prediction of responses to direct-acting antivirals (sofosbuvir + daclatasvir + ribavirin) in HCV genotype-4 patients. METHODS: The serum levels of four miRNAs (miRNA-21, 199, 448 and 181c) were assessed in 150 HCV patients and 50 healthy controls using quantitative real-time PCR. The diagnostic accuracy was determined using receiver operating characteristic (ROC) curve. RESULTS: The four studied miRNAs showed significant upregulation in HCV patients compared with controls. There were significant upregulation of MiR-199 and significant downregulation of miR-448 in late stages of fibrosis with high diagnostic accuracy (area under the curve "AUC" = 0.989%; P < .001) and (AUC = 0.0.672; P > .001), respectively. Regarding response to treatment, only miR-199 showed a significant upregulation in non-responder patients with high diagnostic accuracy (AUC = 0.968; P < .001). CONCLUSION: miR-199 and miR-448 could serve as valuable non-invasive biomarkers for assessment of liver fibrosis progression. Additionally, miR-199 could be also a potential biomarker for assessment of treatment efficacy among HCV patients. Therefore, miR-199 and miR-448 serum levels should be considered during the treatment of HCV genotype-4 patients in Egypt and the world.

Association between Patients' Knowledge and Adherence to Anticoagulants, and Its Effect on Coagulation Control.

BACKGROUND: Warfarin requires strict monitoring by measuring prothrombin time (PT), international normalized ratio (INR), and time in therapeutic range (TTR). Several factors can lead to poor PT/INR control including vitamin K status, medication adherence, knowledge, and quality of life. The present study aims to assess patient adherence to warfarin treatment and its correlation with INR control. METHODS: A cross-sectional study was conducted between October 2017 and January 2018 at Tripoli University Hospital. Data were collected by structured questionnaires which included; demographic and clinical characteristics, the Oral Anticoagulation Knowledge (OAK) test, and the Morisky Medication Adherence Scale (MMAS-8). RESULTS: The final analysis included 88 out of 140 patients (73.33%). There were significant differences in age range, gender, marital status, and education level between the 2 groups (poor knowledge and adequate knowledge) (p < 0.05). There was a significant positive correlation between OAK test score and TTR. Overall, 76.2% of patients were adherent to warfarin (MMAS score ≥6) and 20.45% of patients were of high adherence (MMAS score of 8). The median score was 6 (IQR 6-7). A statistically significant, strong positive correlation between adherence to medication and TTR as an indicator of INR control was found (rs[86] = 0.472, p < 0.0001). CONCLUSION: The study addressed and identified several areas for future improvement of patient outcomes. The implementation of new approaches to enhance patient knowledge and adherence is warranted, and measures to provide treatment for all patients that require it are needed, to improve outcomes and decrease adverse drug effects.

Informed self-assessment versus preceptor evaluation: a comparative study of pediatric procedural skills acquisition of fifth year medical students.

BACKGROUND: Simulation training is widely used in medical education as students rarely perform clinical procedures, and confidence can influence practitioners' ability to perform procedures. Thus, this study assessed students' perceptions and experiences of a pediatric skills program and compared their informed self-assessment with their preceptor-evaluated performance competency for several pediatric clinical procedures. METHODS: A total of 65 final-year medical students attended a weeklong pediatric skills training course by the University of Tripoli that used a manikin and various clinical scenarios to simulate real-life cases. Participants completed questionnaires self-assessing their performance skills, while examiners evaluated each students' competency on five procedural skills (lumbar puncture, nasogastric tube insertion, umbilical vein catheterization, intraosseous access, and suprapubic aspiration) using an objective structured clinical examination (OSCE) model. Differences between agreement levels in question responses were evaluated through a nonparametric chi-square test for a goodness of test fit, and the relationship between confidence levels and the OSCE scores for each procedure was assessed using Spearman's rank-order correlation. RESULTS: All participants completed the informed self-assessment questionnaire and OSCE stations. The frequency differences in agreement levels in students' questionnaire responses were statistically significant. No significant differences were found between students' self-assessment and preceptors' evaluation scores. For each procedure's passing score rate, umbilical vein catheterization had the highest passing rate (78.5%) and nasogastric tube placement the lowest (56.9%). The mean performance scores were above passing for all procedures. The Wilcoxon signed-rank test revealed no significant differences between participants' self-assessment and their preceptor-evaluated competency; students correctly perceived and assessed their ability to perform each procedure. CONCLUSIONS: High competence in several life-saving procedures was demonstrated among final-year medical students. The need for consistent and timely feedback, methods to increase medical students' confidence, and further development and improvement of competency-based assessments are also highlighted.

Symphyseal plate with trans-symphyseal cross-screws for fixation of tile-type B1 pelvic ring injuries: radiological and functional evaluation.

INTRODUCTION: Failure to neutralize the different physiological forces acting on the inferior as well as the superior regions of the pubic symphysis, the long healing time, is blamed for the repeated failures of conventional superior symphyseal plating. MATERIAL AND METHODS: A three years prospective case series study between January 2017 and December 2019 was done, to evaluate the radiological and the functional outcomes, using Matta/Tornetta and Majeed criteria respectively, of the combination of trans-symphyseal cross-screws configuration and superior symphyseal plate in Tile-type B1 pelvic injuries. RESULTS: Thirty patients, 18 with anteroposterior compression type II and 12 with type III, with a mean follow-up of 20 months ±5 were included. Radiologically, 26 (86.6%) cases showed an excellent, one (3.3%) good and three (10%) fair outcome. Clinically, excellent outcome in 26 (86.6%) cases, good in two (6.6%) cases, and fair in two (6.6%). Intra-operative drill bit breakage occurred in three (10%) cases and was the only reported technical complication. Significant re-displacement was reported in three (10%) cases. CONCLUSION: The open trans-symphyseal cross-screws for fixation of the superior symphyseal plate is a simple, efficient, and safe technique with the biomechanical advantages of an extra-fixation point to the inferior symphysis together with a long and a strong bony anchorage.

Treatment of non-united femoral neck fracture by a novel subtrochanteric angulation lateral translation valgus osteotomy (SALVA osteotomy).

PURPOSE: The ideal treatment of non-united and neglected fracture neck femur in the young adult still remains unclear and is characterized by many biological and biomechanical challenges. METHODS: Twenty-one patients with non-united or neglected fracture neck femur aged between 19 and 50 years were treated by a novel subtrochanteric valgus osteotomy and were followed up for a mean of 26.7 months. Patients were assessed by radiological parameters, the Harris Hip Score, Oxford Knee Score, and Askin Bryan Criteria to categorize the overall outcome of the patients at 24 months. Other outcome measures included the occurrence of AVN, adductor lever arm, leg length discrepancy, and mechanical implant failure. RESULTS: All patients treated with the SALVA osteotomy consolidated and displayed a marked improvement of functional and radiological outcome measures. Nevertheless, there were 2 mechanical failures in patients with marked osteopenia and three developed AVN. CONCLUSIONS: In patients with un-united/neglected fracture neck femur, SALVA osteotomy appears to be reliable and reproducible. It also restores the abductor lever arm and improves the leg length discrepancy. Technically less demanding conversion to arthroplasty remains still possible prospectively.

Neuroimaging with Radiopharmaceuticals Targeting the Glutamatergic System.

Radiopharmacy at ETH has worked on the development of novel PET tracers for neuro-, cardiac- and tumor imaging for many years. In this paper, our efforts on targeting the glutamatergic system of the metabotropic glutamate receptor subtype 5 (mGluR5) and the ionotropic N-methyl-D-aspartate (NMDA) receptor are summarized. We briefly described the principles of positron emission tomography (PET) tracer development for the central nervous system (CNS) and the radiolabeling methods used in our laboratory. To assess the radioligands, results of in vitro autoradiography, biodistribution, and metabolite studies as well as PET imaging data are discussed. Furthermore, key PET parameters for kinetic modeling and quantification methods are provided. Two mGluR5 PET tracers, [11C]ABP688 and [18F]PSS232, were translated in our GMP labs and evaluated in human subjects. The newly developed GluN2B PET tracer [11C]Me-NB1 is currently being investigated in a first-in-human PET study and several F-18 labeled tracers are being evaluated in non-human primates in which the first-in-class will be translated for human studies.

An Integrative Proteomic Approach Identifies Novel Cellular SMYD2 Substrates.

Protein methylation is a post-translational modification with important roles in transcriptional regulation and other biological processes, but the enzyme-substrate relationship between the 68 known human protein methyltransferases and the thousands of reported methylation sites is poorly understood. Here, we propose a bioinformatic approach that integrates structural, biochemical, cellular, and proteomic data to identify novel cellular substrates of the lysine methyltransferase SMYD2. Of the 14 novel putative SMYD2 substrates identified by our approach, six were confirmed in cells by immunoprecipitation: MAPT, CCAR2, EEF2, NCOA3, STUB1, and UTP14A. Treatment with the selective SMYD2 inhibitor BAY-598 abrogated the methylation signal, indicating that methylation of these novel substrates was dependent on the catalytic activity of the enzyme. We believe that our integrative approach can be applied to other protein lysine methyltransferases, and help understand how lysine methylation participates in wider signaling processes.

Structural Basis for the Discriminative Recognition of N6-Methyladenosine RNA by the Human YT521-B Homology Domain Family of Proteins.

N(6)-Methyladenosine (m(6)A) is the most abundant internal modification in RNA and is specifically recognized by YT521-B homology (YTH) domain-containing proteins. Recently we reported that YTHDC1 prefers guanosine and disfavors adenosine at the position preceding the m(6)A nucleotide in RNA and preferentially binds to the GG(m(6)A)C sequence. Now we systematically characterized the binding affinities of the YTH domains of three other human proteins and yeast YTH domain protein Pho92 and determined the crystal structures of the YTH domains of human YTHDF1 and yeast Pho92 in complex with a 5-mer m(6)A RNA, respectively. Our binding and structural data revealed that the YTH domain used a conserved aromatic cage to recognize m(6)A. Nevertheless, none of these YTH domains, except YTHDC1, display sequence selectivity at the position preceding the m(6)A modification. Structural comparison of these different YTH domains revealed that among those, only YTHDC1 harbors a distinctly selective binding pocket for the nucleotide preceding the m(6)A nucleotide.

Association of treatment satisfaction and physician trust with glycemic control among primary care patients with type 2 diabetes in Egypt.

Objectives: To evaluate the association of diabetes treatment satisfaction and trust in family physicians with glycemic control among primary care patients with type 2 diabetes mellitus. Methods: A cross-sectional study on 319 patients with type 2 diabetes mellitus from five primary healthcare centers in Egypt. Data were collected from February to August 2021 using a structured questionnaire that contained six parts: sociodemographic data, disease profile, the Diabetes Treatment Satisfaction Questionnaire (DTSQ), 8-item Morisky Medication Adherence Scale (MMAS-8), self-reported medication knowledge questionnaire (MKQ), and revised healthcare relationship trust scale (HCR). Multiple linear regression analysis was used to assess predictors of treatment satisfaction, physician trust, and HbA1c level. P values less than 0.05 were considered significant. Results: The mean age was 59.66 years (± 7.87 years) and 55.17% were females. Multiple linear regression analysis for predicting HbA1c showed that HbA1c level was lower in patients with higher treatment satisfaction scores (ß = - 0.289, p < 0.001) and higher medication adherence scores (ß = - 0.198, p = 0.001). Treatment satisfaction scores were positively predicted by higher physician trust scores (ß = 0.301, p < 0.001), increased medication adherence scores (ß = 0.160, p = 0.002), and longer duration of diabetes (ß = 0.226, p < 0.001). Positive predictors for physician trust included HbA1c level (ß = 0.141, p = 0.012), medication knowledge (ß = 0.280, p < 0.001), diabetes treatment satisfaction (ß = 0.366, p < 0.001) and medication adherence (ß = 0.146, p = 0.011). Conclusion: Optimizing diabetes treatment satisfaction and physician trust could have favorable associations with medication adherence and medication knowledge with a possible improvement in glycemic control. Family physicians should incorporate patients reported outcomes alongside traditional clinical measures in evaluating diabetes management in primary care.

Health coaching for hypertension control in primary care patients with uncontrolled hypertension in Egypt.

BACKGROUND: Health coaching effectively improves hypertension self-care activities and the control of blood pressure (BP) in hypertensive patients. Studies on the effects of health coaching on patients in primary care with uncontrolled hypertension in developing countries are limited. In this study, the effectiveness of health coaching on hypertension self-care and BP control was assessed in patients who have uncontrolled hypertension compared to standard care in Egypt. MATERIALS AND METHODS: Our quasi-experimental study included control and intervention groups. The intervention group included 70 participants who received health coaching sessions (face-to-face and by telephone) besides the standard care, whereas the control group included 71 participants who only received the standard care. The study was conducted between July 2020 and November 2021. The participants were recruited from three primary healthcare settings in the Port Said Governorate. Personal and medical history, BP measurements, and hypertension self-care activity level effects (H-SCALE) were obtained. Paired-t-test was used to assess the changes in BP measurement, and H-SCALE score before and after receiving the health coaching. McNemar's test was used to assess changes in controlled BP and optimal hypertension self-care activities between control and health coached groups. Multiple logistic regression analysis assessed the predictors of better BP control. RESULTS: Health coaching resulted in more controlled BP (51.4%, P < 0.001) compared to the delivery of only usual care (11.3%, P = 0.008). The intervention showed a significant promotion in hypertension self-care activities, including medication usage (P < 0.001), low-salt diet (P < 0.001), and weight management (P < 0.001). The H-SCALE score mean change was the only predictor for BP control (odds ratio 1.057, P = 0.048) in the intervention group after 6 months. CONCLUSION: Intervention including traditional health coaching and phone calls is a beneficial modality for the promotion of hypertension self-care and improvement of BP control in primary care patients with uncontrolled hypertension.

The Cytotoxic Effect of Ciprofloxacin Laetrile Combination on Esophageal Cancer Cell Line.

OBJECTIVE: aim of this study was to examine the synergistic effect between the antibacterial drug ciprofloxacin and the natural compound laetrile on esophageal cancer cells, specifically focusing on their combined cytotoxic effect. METHODS: The combined cytotoxic effects of two alternative incubation durations (24 and 72 hours) were studied using an esophageal cancer cell line.  Ciprofloxacin, laetrile, and their combinations were tested at concentrations ranging from 1 to 1000 micrograms/milliliter, to enhance the safety of the combination, the concentrations of the combination constituents were reduced by half compared to when they are used individually, the combination index was then calculated to estimate the components' possible synergistic effects. RESULT: The results indicate that the combined cytotoxicity of ciprofloxacin and laetrile was greater than the cytotoxicity of either ciprofloxacin or laetrile alone, the combination cytotoxicity increased with higher concentrations and longer incubation periods, in other words, the cytotoxicity pattern of the combination was time-dependent (cell-cycle specific), and concentration dependent, (cell-cycle non-specific). CONCLUSION: The study found that the combination of ciprofloxacin and laetrile had a greater inhibitory effect on the growth of esophageal cancer cells compared to ciprofloxacin or laetrile alone. This suggests a synergistic effect between the components of the mixture, which can be attributed to a complementary mechanism between the ingredients in the combination.