High UV damage and low repair, but not cytosine deamination, stimulate mutation hotspots at ETS binding sites in melanoma.

Noncoding mutation hotspots have been identified in melanoma and many of them occur at the binding sites of E26 transformation-specific (ETS) proteins; however, their formation mechanism and functional impacts are not fully understood. Here, we used UV (Ultraviolet) damage sequencing data and analyzed cyclobutane pyrimidine dimer (CPD) formation, DNA repair, and CPD deamination in human cells at single-nucleotide resolution. Our data show prominent CPD hotspots immediately after UV irradiation at ETS binding sites, particularly at sites with a conserved TTCCGG motif, which correlate with mutation hotspots identified in cutaneous melanoma. Additionally, CPDs are repaired slower at ETS binding sites than in flanking DNA. Cytosine deamination in CPDs to uracil is suggested as an important step for UV mutagenesis. However, we found that CPD deamination is significantly suppressed at ETS binding sites, particularly for the CPD hotspot on the 5' side of the ETS motif, arguing against a role for CPD deamination in promoting ETS-associated UV mutations. Finally, we analyzed a subset of frequently mutated promoters, including the ribosomal protein genes RPL13A and RPS20, and found that mutations in the ETS motif can significantly reduce the promoter activity. Thus, our data identify high UV damage and low repair, but not CPD deamination, as the main mechanism for ETS-associated mutations in melanoma and uncover important roles of often-overlooked mutation hotspots in perturbing gene transcription.

The interplay of 3D genome organization with UV-induced DNA damage and repair.

The 3D organization of the eukaryotic genome is crucial for various cellular processes such as gene expression and epigenetic regulation, as well as for maintaining genome integrity. However, the interplay between UV-induced DNA damage and repair with the 3D structure of the genome is not well understood. Here, we used state-of-the-art Hi-C, Damage-seq, and XR-seq datasets and in silico simulations to investigate the synergistic effects of UV damage and 3D genome organization. Our findings demonstrate that the peripheral 3D organization of the genome shields the central regions of genomic DNA from UV-induced damage. Additionally, we observed that potential damage sites of pyrimidine-pyrimidone (6-4) photoproducts are more prevalent in the nucleus center, possibly indicating an evolutionary pressure against those sites at the periphery. Interestingly, we found no correlation between repair efficiency and 3D structure after 12 min of irradiation, suggesting that UV radiation alters the genome's 3D organization in a short period of time. Interestingly, however, 2 h after UV induction, we observed more efficient repair levels in the center of the nucleus relative to the periphery. These results have implications for understanding the etiology of cancer and other diseases, as the interplay between UV radiation and the 3D genome may play a role in the development of genetic mutations and genomic instability.

Cpd-A1 alleviates acute kidney injury by inhibiting ferroptosis.

Acute kidney injury (AKI) is defined as sudden loss of renal function characterized by increased serum creatinine levels and reduced urinary output with a duration of 7 days. Ferroptosis, an iron-dependent regulated necrotic pathway, has been implicated in the progression of AKI, while ferrostatin-1 (Fer-1), a selective inhibitor of ferroptosis, inhibited renal damage, oxidative stress and tubular cell death in AKI mouse models. However, the clinical translation of Fer-1 is limited due to its lack of efficacy and metabolic instability. In this study we designed and synthesized four Fer-1 analogs (Cpd-A1, Cpd-B1, Cpd-B2, Cpd-B3) with superior plasma stability, and evaluated their therapeutic potential in the treatment of AKI. Compared with Fer-1, all the four analogs displayed a higher distribution in mouse renal tissue in a pharmacokinetic assay and a more effective ferroptosis inhibition in erastin-treated mouse tubular epithelial cells (mTECs) with Cpd-A1 (N-methyl-substituted-tetrazole-Fer-1 analog) being the most efficacious one. In hypoxia/reoxygenation (H/R)- or LPS-treated mTECs, treatment with Cpd-A1 (0.25 µM) effectively attenuated cell damage, reduced inflammatory responses, and inhibited ferroptosis. In ischemia/reperfusion (I/R)- or cecal ligation and puncture (CLP)-induced AKI mouse models, pre-injection of Cpd-A1 (1.25, 2.5, 5 mg·kg-1·d-1, i.p.) dose-dependently improved kidney function, mitigated renal tubular injury, and abrogated inflammation. We conclude that Cpd-A1 may serve as a promising therapeutic agent for the treatment of AKI.

Access to continuous professional development for capacity building among nurses and midwives providing emergency obstetric and neonatal care in Rwanda.

BACKGROUND: Nurses and midwives are at the forefront of the provision of Emergency Obstetric and Neonatal Care (EmONC) and Continuous Professional Development (CPD) is crucial to provide them with competencies they need to provide quality services. This research aimed to assess uptake and accessibility of midwives and nurses to CPD and determine their knowledge and skills gaps in key competencies of EmONC to inform the CPD programming. METHODS: The study applied a quantitative, cross-sectional, and descriptive research methodology. Using a random selection, forty (40) health facilities (HFs) were selected out of 445 HFs that performed at least 20 deliveries per month from July 1st, 2020 to June 30th, 2021 in Rwanda. Questionnaires were used to collect data on updates of CPD, knowledge on EmONC and delivery methods to accessCPD. Data was analyzed using IBM SPSS statistics 27 software. RESULTS: Nurses and midwives are required by the Rwandan midwifery regulatory body to complete at least 60 CPD credits before license renewal. However, the study findings revealed that most health care providers (HCPs) have not been trained on EmONC after graduation from their formal education. Results indicated that HCPs who had acquired less than 60 CPD credits related to EmONC training were 79.9% overall, 56.3% in hospitals, 82.2% at health centres and 100% at the health post levels. This resulted in skills and knowledge gaps in management of Pre/Eclampsia, Postpartum Hemorrhage and essential newborn care. The most common method to access CPD credits included workshops (43.6%) and online training (34.5%). Majority of HCPs noted that it was difficult to achieve the required CPD credits (57.0%). CONCLUSION: The findings from this study revealed a low uptake of critical EmONC training by nurses and midwives in the form of CPD. The study suggests a need to integrate EmONC into the health workforce capacity building plan at all levels and to make such training systematic and available in multiple and easily accessible formats. IMPLICATION ON NURSING AND MIDWIFERY POLICY: Findings will inform the revision of policies and strategies to improve CPD towards accelerating capacity for the reduction of preventable maternal and perinatal deaths as well as reducing maternal disabilities in Rwanda.

Identifying training needs of practising community pharmacists in Jordan-a self-assessment study.

BACKGROUND: Being the professional membership body for pharmacists in Jordan, the Jordan Pharmacists Association (JPA) took the initiative to establish a training centre for practising pharmacists. This study aims to identify the self-assessed training priorities of community pharmacists in Jordan. METHODS: In the period between August and October 2022, an online self-administered questionnaire was distributed using a variety of participants' identification and recruitment approaches. The questionnaire targeted currently practising community pharmacists. Data were analysed descriptively and inferentially. RESULTS: In total, 470 community pharmacists participated in this study. Of 470 participants, 307 (65.3%) were employees, of which 206 were full-time employees. Results showed that only 97 (21%) had access to an in-house training programme or scheme. Self-assessment of training needs highlighted differences between the three competencies clusters. While administrative and managerial skills and competencies were more frequently prioritised on average than the other two clusters, interpersonal and communication skills were needed the least. Evidence showed a significant difference between female and male participants regarding the need for training addressing maternity and early childhood health training issues. Lastly, the role-based comparison showed that, compared to pharmacy owners, employees had a significantly higher need for training related to bookkeeping and taxation returns preparation and how to handle and manage records of narcotic and controlled medicines. CONCLUSIONS: If training and development programmes are tailored to address specific needs in administrative, clinical, and interpersonal competencies, community pharmacists have the potential to enhance public health, expand their role, provide patient-centred care, and support the national healthcare system.

Beyond residency: 'The imperative of lifelong learning in medical practice'.

The CPD landscape is rapidly evolving and may be affecting patient outcomes. This also poses challenges to healthcare professionals, some of whom are experiencing a lot of stress leading to burnout. Medical and residency training are very structured and the importance of CPD occurring in a non-structured setting to prevent professional stagnation cannot be overemphasized. The need for lifelong learning post- residency, which encompasses a much longer period until retirement, is underscored hence the need for a cultural shift. Adult learning needs emphasis and a focus on addressing individual needs is required. Reasons for physician reluctance and inertia towards CPD such as lack of time, self-sufficiency, occasional journal reading, and skepticism need ongoing exploration. Comprehensive measures, including integrated EMR designs, personal learning projects, practice updates, reward systems, physician surveys, equal weight for formal and informal CPD, and promoting a learning mindset are proposed.

Medical education in Georgia.

Since 1991, there have been significant changes in medical education in Georgia. Key changes include adapting national legislation toward international standards, establishing the National Center for Education Quality Enhancement (NCEQE), which was recognized in 2018 by the World Federation for Medical Education (WFME) as an accrediting agency and opening the Association for Medical Education in Europe (AMEE) International Networking Center in 2019. Undergraduate medical education, regulated by the Ministry of Education, Science and Youth of Georgia, spans six years. MD graduates then have options for further career paths, including working as junior doctors, residency, and/or pursuing PhD research.The main challenges the country presently faces are:the need to reduce the increasing number of (mainly) private medical schools. Recent updates to the national standards for undergraduate medical education have imposed stricter accreditation requirements for MD programs, resulting in the closure of schools that fail to meet these standards;postgraduate medical education is governed by the Ministry of Internally Displaced Persons from the Occupied Territories, Labor, Health and Social Affairs of Georgia (MOH) and needs further reform due to limited and paid residency positions;continuous professional development (CPD) was optional until recently, which led to an increase in professional inaccuracy and malpractice cases. To address this, regulatory bodies, including the MOH and professional associations, are preparing the legal basis for introducing compulsory CPD.

An overview of global CME/CPD systems.

PURPOSE: Engagement in CME/CPD has a positive impact on healthcare professionals' (HCPs) knowledge, skills, and performance, and on patient outcomes, therefore it is critical to better understand the components of CME/CPD systems that foster engagement, high-quality education, and impact. METHODS: An assessment of CME/CPD systems was conducted using a mixed-methods approach that included interviews with in-country subject matter experts and qualitative and quantitative data from practicing in-country physicians. RESULTS: Results demonstrate areas of consistency in CME/CPD systems across world regions that included: types of educational providers; types of credit; educational formats; self-tracking of participation; high-degree of compliance when education is mandatory; overall satisfaction with available education; strong support for interprofessional education; and lack of alignment or evaluation of engagement in education with population health outcomes. Areas of variation included: whether engagement in education is required as a condition to practice medicine; whether regulations are uniformly applied; if mechanisms to ensure independence existed; and physician perceptions of independence. CONCLUSION: Results of this assessment maybe used by a variety of different stakeholders to assess how well country-level CME/CPD systems are meeting the needs of practicing physicians and determine what, if any, changes might need to be implemented to improve outcomes.

Knowledge, usability and challenges of e-learning platforms for continuing Professional Development of healthcare professionals at University Teaching Hospital of Kigali.

BACKGROUND: Healthcare professionals constitute a critical component of clinical care services. To provide the expected service, they must continuously develop their profession through continuous learning. This kind of learning is recognized as continuing professional development (CPD). Traditionally, CPD is offered onsite. Onsite training is associated with some barriers that prevent healthcare professionals from attending such educational activities, including financial difficulties and long distance. This is why online learning is proposed to overcome these barriers. OBJECTIVE: The main purpose was to evaluate usability, knowledge and challenges of e-learning platforms for CPD of healthcare professionals at University Teaching Hospital of Kigali (CHUK). METHODS: The cross-sectional quantitative study approach was utilized; the data was collected at the workplace of nurses, midwives, and allied health professionals by using a pre-designed questionnaire. The data were analyzed using Statistical Package for the Social Sciences (SPSS) version 25 and presented as frequencies. RESULTS: A significant majority was aware of CPD e-learning platforms. For example, 95.7% of the participants were familiar with these platforms, indicating that they had some degree of knowledge about their existence and purpose. Regarding the mode of accessing CPD courses, 82.1% of participants preferred online platforms, demonstrating a strong will to use e-learning platforms. CONCLUSION: This study highlighted a high level of awareness and utilization of CPD e-learning platforms among healthcare professionals at CHUK, additionally, participants expressed confidence in using the platforms but emphasized the need for further support and training.

A scoping review of continuing education models and statutory requirements for pharmacists globally.

BACKGROUND: In the dynamic field of pharmacy amongst a diverse array of countries with disparate income levels, pharmacists play a pivotal role in integrating emerging scientific knowledge into their practice while adapting to evolving therapeutic interventions and expanding service delivery responsibilities. Lifelong Learning (LLL) is cultivated through continuing professional education (CPE) and continuing professional development (CPD), indispensable components ensuring sustained professional competence and heightened patient care quality. The global landscape witnesses diverse LLL activities tailored to pharmacists' learning needs and preferences. This scoping review maps and synthesises a comprehensive global perspective on the existing knowledge regarding CPE/CPD models, statutory requirements, and pharmacists' preferences for LLL activities. OBJECTIVE: To comprehensively investigate global models of CPE/CPD for pharmacists' and examine the statutory requirements governing pharmacists' registration and licensure. METHOD: A literature search of PubMed, Google Scholar, Web of Science, and the University of KwaZulu-Natal library search engine was undertaken for studies between January 2012 and February 2023. The article selection and reporting followed the recommendations made by PRISMA (Preferred Reporting Items of Systematic Reviews and Meta-Analyses) guidelines. The articles were tabulated based on their respective country's income level, continuing education models employed, country-specific statutory requirements, and pharmacists' preferences for LLL activities. RESULTS: Of the initial 3974 publications identified through the database search, 24 studies met the review criteria. The majority of the articles originated from high-income countries (HICs) (14/24, 58.3%), and most employed the mandatory CPD points system (21/24, 87.5%). However, in some HICs and upper-middle income countries (UMICs), the CPE/CPD is non-mandatory. While most countries (19/24, 79.2%) offer various LLL formats, the preference of pharmacists remains primarily face-to-face learning (13/24, 54.2%). However, workplace learning (3/24, 12.5%) and blended learning (7/24, 29.1%) are mentioned in some studies. CONCLUSION: Diverse models of CPE/CPD alongside statutory requirements persist globally and evolve, shaped by varied implementation experiences. HICs lead in CPD models, while the implementation in low- and middle-income countries (LMICs) and low-income countries (LICs) requires further exploration for inclusivity and effectiveness. A few UMICs are either initiating or in early stages of implementing the CPD models. Structured planning for LLL activities is increasingly a global requirement for pharmacists' licensure. The essential progression of pharmacy practice in developing healthcare systems necessitates a mandatory CPD model. Ongoing research is crucial to fortify the implementation, align and unify the CPD model with evolving pharmacy profession needs.

Evaluating the impact of the supporting the advancement of research skills (STARS) programme on research knowledge, engagement and capacity-building in a health and social care organisation in England.

OBJECTIVES: To evaluate the impact a novel education programme - to improve research engagement, awareness, understanding and confidence - had on a diverse health and social care workforce. Barriers and facilitators to engagement were explored together with research capacity-building opportunities and ways to embed a research culture. The programme is entitled 'Supporting The Advancement of Research Skills' (STARS programme); the paper reports findings from a health and social care setting in England, UK. METHODS: A four-level outcome framework guided the approach to evaluation and was further informed by key principles of research capacity development and relevant theory. Quantitative data were collected from learners before and after engagement; these were analysed descriptively. Semi-structured online interviews were conducted with learners and analysed thematically. A purposive sample was achieved to include a diversity in age, gender, health and social care profession, and level of attendance (regular attendees, moderate attendees and non-attenders). RESULTS: The evaluation spanned 18 half-day workshops and 11 seminars delivered by expert educators. 165 (2% of total staff at Midlands Partnership University NHS Foundation Trust (MPFT)) staffs booked one or more education sessions; 128 (77%) including Allied Health Professionals (AHPs), psychologists, nursing and midwifery, and social workers attended one or more session. Key themes of engagement with teaching sessions, relevance and impact of training and promoting a research active environment were identified with relevant sub-themes. Positive impacts of training were described in terms of research confidence, intentions, career planning and application of research skills as a direct result of training. Lack of dedicated time for research engagement, work pressures and time commitments required for the programme were key barriers. Facilitators that facilitated engagement are also described. CONCLUSIONS: Findings demonstrate the impact that a free, virtual and high-quality research education programme had at individual and organisational levels. The programme is the product of a successful collaboration between health and social care and academic organisations; this provides a useful framework for others to adapt and adopt. Key barriers to attendance and engagement spoke to system-wide challenges that an education programme could not address in the short-term. Potential solutions are discussed in relation to protecting staff time, achieving management buy-in, recognising research champions, and having a clear communication strategy.

Variable Inhibition of DNA Unwinding Rates Catalyzed by the SARS-CoV-2 Helicase Nsp13 by Structurally Distinct Single DNA Lesions.

The SARS-CoV-2 helicase, non-structural protein 13 (Nsp13), plays an essential role in viral replication, translocating in the 5' → 3' direction as it unwinds double-stranded RNA/DNA. We investigated the impact of structurally distinct DNA lesions on DNA unwinding catalyzed by Nsp13. The selected lesions include two benzo[a]pyrene (B[a]P)-derived dG adducts, the UV-induced cyclobutane pyrimidine dimer (CPD), and the pyrimidine (6-4) pyrimidone (6-4PP) photolesion. The experimentally observed unwinding rate constants (kobs) and processivities (P) were examined. Relative to undamaged DNA, the kobs values were diminished by factors of up to ~15 for B[a]P adducts but only by factors of ~2-5 for photolesions. A minor-groove-oriented B[a]P adduct showed the smallest impact on P, which decreased by ~11% compared to unmodified DNA, while an intercalated one reduced P by ~67%. However, the photolesions showed a greater impact on the processivities; notably, the CPD, with the highest kobs value, exhibited the lowest P, which was reduced by ~90%. Our findings thus show that DNA unwinding efficiencies are lesion-dependent and most strongly inhibited by the CPD, leading to the conclusion that processivity is a better measure of DNA lesions' inhibitory effects than unwinding rate constants.

1,25-Dihydroxyvitamin D3 Suppresses UV-Induced Poly(ADP-Ribose) Levels in Primary Human Keratinocytes, as Detected by a Novel Whole-Cell ELISA.

Photoprotective properties of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) to reduce UV-induced DNA damage have been established in several studies. UV-induced DNA damage in skin such as single or double strand breaks is known to initiate several cellular mechanisms including activation of poly(ADP-ribose) (pADPr) polymerase-1 (PARP-1). DNA damage from UV also increases extracellular signal-related kinase (ERK) phosphorylation, which further increases PARP activity. PARP-1 functions by using cellular nicotinamide adenine dinucleotide (NAD+) to synthesise pADPr moieties and attach these to target proteins involved in DNA repair. Excessive PARP-1 activation following cellular stress such as UV irradiation may result in excessive levels of cellular pADPr. This can also have deleterious effects on cellular energy levels due to depletion of NAD+ to suboptimal levels. Since our previous work indicated that 1,25(OH)2D3 reduced UV-induced DNA damage in part through increased repair via increased energy availability, the current study investigated the effect of 1,25(OH)2D3 on UV-induced PARP-1 activity using a novel whole-cell enzyme- linked immunosorbent assay (ELISA) which quantified levels of the enzymatic product of PARP-1, pADPr. This whole cell assay used around 5000 cells per replicate measurement, which represents a 200-400-fold decrease in cell requirement compared to current commercial assays that measure in vitro pADPr levels. Using our assay, we observed that UV exposure significantly increased pADPr levels in human keratinocytes, while 1,25(OH)2D3 significantly reduced levels of UV-induced pADPr in primary human keratinocytes to a similar extent as a known PARP-1 inhibitor, 3-aminobenzamide (3AB). Further, both 1,25(OH)2D3 and 3AB as well as a peptide inhibitor of ERK-phosphorylation significantly reduced DNA damage in UV-exposed keratinocytes. The current findings support the proposal that reduction in pADPr levels may be critical for the function of 1,25(OH)2D3 in skin to reduce UV-induced DNA damage.

Evaluation of factors affecting pharmacists and pharmacy technicians' satisfaction towards practicing CE activities in Saudi Arabia.

Background: Continuing education (CE) is an essential requirement for pharmacy professionals to stay abreast with the evolving knowledge and skills of the practice and meet the regulatory mandate. The purpose of this research is to assess factors affecting the satisfaction of pharmacists and pharmacy technicians towards CE practices in Saudi Arabia. Material and methods: A self-administered survey instrument was developed following an extensive literature search. The questionnaire consisted of three sections: participants' demographics, data on CE activities over the past year and overall satisfaction, and statements of barriers (14 items) and facilitators (12 items) for participation in CE activities (scored on a 5-point Likert scale (5 = always, 1 = never)). The survey was piloted and then distributed as a link through the Saudi Commission for Health Specialties and Saudi Pharmaceutical Society (SPS) between Jan 2018 and Feb 2019. Results: Data was available on 398 pharmacists and 40 pharmacy technicians (completion rate was 55 %). The majority were practitioners, male, working in a hospital setting and had more than five years of practice experience. Half of the participants were from the Central Region and about one-third were non-Saudi. Only a quarter of the participants were satisfied/very satisfied with the current CE practices in Saudi Arabia. Job constraints (62.7 %), cost (55.9 %), schedule of CE activities (55.4 %), lack of information on CE opportunities (53 %) and professional burnout (49.7 %) were the top barriers. There was a significant level of dissatisfaction among pharmacy technicians when compared to pharmacists (p = 0.003), as well as among Saudi pharmacists when compared to non-Saudi pharmacists (p = 0.002). Lack of relevant CE activities (p = 0.05), lack of quality activities (p = 0.002), lack of recognition (p = 0.013) and lack of internet access (p = 0.006) were significantly more barriers for pharmacy technicians compared to pharmacists. The most identified facilitators to engage in CE activities were a personal desire to learn (78.4 %), the requirement to maintain a professional license (73.8 %) and relaxation provided by learning (58.5 %) and networking opportunities (53.4 %). The majority of the participants preferred conferences or interactive workshops, short CE over half a day or less, and the topic of disease management/drug therapy. Conclusion: The findings of the study highlight the need for a partnership strategy that includes various stakeholders to improve CE program quality and accessibility that supports and promotes the professional development of pharmacists and pharmacy technicians in Saudi Arabia. It also underscores the importance of meeting the preferences of pharmacy practitioners when designing CE programs and aligning such activities with their practices.

Autophagy modulators influence the content of important signalling molecules in PS-positive extracellular vesicles.

Extracellular vesicles (EVs) are important mediators of intercellular communication in the tumour microenvironment. Many studies suggest that cancer cells release higher amounts of EVs exposing phosphatidylserine (PS) at the surface. There are lots of interconnections between EVs biogenesis and autophagy machinery. Modulation of autophagy can probably affect not only the quantity of EVs but also their content, which can deeply influence the resulting pro-tumourigenic or anticancer effect of autophagy modulators. In this study, we found that autophagy modulators autophinib, CPD18, EACC, bafilomycin A1 (BAFA1), 3-hydroxychloroquine (HCQ), rapamycin, NVP-BEZ235, Torin1, and starvation significantly alter the composition of the protein content of phosphatidylserine-positive EVs (PS-EVs) produced by cancer cells. The greatest impact had HCQ, BAFA1, CPD18, and starvation. The most abundant proteins in PS-EVs were proteins typical for extracellular exosomes, cytosol, cytoplasm, and cell surface involved in cell adhesion and angiogenesis. PS-EVs protein content involved mitochondrial proteins and signalling molecules such as SQSTM1 and TGFß1 pro-protein. Interestingly, PS-EVs contained no commonly determined cytokines, such as IL-6, IL-8, GRO-α, MCP-1, RANTES, and GM-CSF, which indicates that secretion of these cytokines is not predominantly mediated through PS-EVs. Nevertheless, the altered protein content of PS-EVs can still participate in the modulation of the fibroblast metabolism and phenotype as p21 was accumulated in fibroblasts influenced by EVs derived from CPD18-treated FaDu cells. The altered protein content of PS-EVs (data are available via ProteomeXchange with identifier PXD037164) also provides information about the cellular compartments and processes that are affected by the applied autophagy modulators. Video Abstract.

Identification of a Noxo1 inhibitor by addition of a polyethylene glycol chain.

Reactive oxygen species (ROS) are a heterogeneous group of highly reactive ions and molecules derived from molecular oxygen (O2) which can cause DNA damage and lead to skin cancer. NADPH oxidase 1 (Nox1) is a major producer of ROS in the skin upon exposure to ultraviolet light. Functionally, Nox1 forms a holoenzyme complex that generates two superoxide molecules and reduces NADPH. The signaling activation occurs when the organizer subunit Noxo1 translocates to the plasma membrane bringing a cytochrome p450, through interaction with Cyba. We propose to design inhibitors that prevent Cyba-Noxo1 binding as a topical application to reduce UV-generated ROS in human skin cells. Design started from an apocynin backbone structure to generate a small molecule to serve as an anchor point. The initial compound was then modified by addition of a polyethylene glycol linked biotin. Both inhibitors were found to be non-toxic in human keratinocyte cells. Further in vitro experiments using isothermal calorimetric binding quantification showed the modified biotinylated compound bound Noxo1 peptide with a KD of 2 nM. Both using isothermal calorimetric binding and MALDI (TOF) MS showed that binding of a Cyba peptide to Noxo1 was blocked. In vivo experiments were performed using donated skin explants with topical application of the two inhibitors. Experiments show that ultraviolet light exposure of with the lead compound was able to reduce the amount of cyclobutene pyrimidine dimers in DNA, a molecule known to lead to carcinogenesis. Further synthesis showed that the polyethylene glycol but not the biotin was essential for inhibition.

Evaluation of the Diagnostic Value of Cell Population Data in Sepsis in Comparison to Localized Infection, Chronic Inflammation, and Noninfectious Inflammation Cases.

INTRODUCTION: Sepsis, defined as an increase of 2 points or more in the sequential organ failure assessment score, is a life-threatening organ dysfunction caused by the dysregulated host response to infection. Volume-conductivity-scatter (VCS) parameters of cell counters which are known as cell population data (CPD) have been suggested to be beneficial in diagnosing sepsis. We aimed to evaluate the diagnostic value of CPD parameters in sepsis in comparison to nonsystemic infection cases (NSI) and non-infectious acute and chronic inflammatory conditions. MATERIALS AND METHODS: We prospectively included four groups of patients" data: sepsis (n = 66), localized infection (pneumonia, n = 59), chronic inflammation (rheumatoid arthritis, n = 92) and noninfectious inflammation (coronary artery bypass graft operation, n = 56) groups, according to their clinical status and laboratory results. Samples for cell counting and serum markers were collected on the same day of culture collection. VCS parameters were measured by Unicel DxH800 Coulter Cellular Analyzer (Beckman Coulter, USA). RESULTS: Mean neutrophil volume (MN-V-NE), was highest in the sepsis group [155(149-168)] compared to the localized infection [148(140-158)], chronic inflammation [144.5(142-149)] and noninfectious inflammation [149(145.2-153.7)] (P = 0.001, P < 0.001, P < 0.001, respectively). Neutrophil volume SD (SD-V-NE) was higher in the sepsis [21(18.8-23.7)], significantly differentiating sepsis from other groups. The area under curves of procalcitonin and hs-C-reactive protein were 0.846 and 0.837, respectively, in the receiver-operating characteristic curves (ROC) . CPD combinations, (SD-V NE + SD-V LY + SD-V MO), (SD-V NE + SD-V MO), and (MN-V NE + SD-V NE + SD-C LY + SD-V MO) had greater AUC values than procalcitonin's. CONCLUSION: VCS parameters might be promising for differentiating sepsis and non-sepsis cases. Additionally, obtaining these data routinely makes their prospects promising without any additional cost and time.

Continuing professional development (CPD) training needs assessment for medical laboratory professionals in Ethiopia.

BACKGROUND: Continuing professional development (CPD) is required for health workers in practice to update knowledge and skills regularly to match the changing complexity of healthcare needs. The objective of this study was to identify the training needs of Medical Laboratory professionals in Ethiopia. METHODS: A total of 457 medical laboratory professionals from five regions and two city administrations were involved in the study. Data were collected from August 02 to 21, 2021 with structured self-administered online tool with five-point Likert scale. The tool had consent, demography, cross-cutting issues, and main activity area specific to medical laboratory. RESULTS: Majority of the participants were male (80.1%). Participants from Amhara region 110 (24.1%) were the largest groups in the survey followed by Oromia 105 (23%) and Addis Ababa 101 (22.1%). The study participants comprised 54.7% with a bachelor's degree, 31.3% with a diploma (associate degree), and 14% with a master's degree. The participants had varying years of service, ranging from less than one year to over 10 years of experience. Most of the participants work as generalists (24.1%) followed by working in microbiology (17.5%) and parasitology (16%). The majority (96.9%) were working in a public sector or training institutions and the rest were employed in the private sector. Our study showed that the three most important topics selected for training in the cross-cutting health issues were health and emerging technology, computer skills and medico-legal issues. Topics under microbiology, clinical chemistry and molecular diagnostics were selected as the most preferred technical areas for training. Participants have also selected priority topics under research skill and pathophysiology. When the laboratory specific issues were regrouped based on areas of application as technical competence, research skill and pathophysiology, thirteen topics under technical competence, four topics under research skill and three topics under pathophysiology were picked as priority areas. CONCLUSION: In conclusion, our study identified that CPD programs should focus on topics that address technical competence in microbiology, clinical chemistry and molecular diagnostics. Additionally competencies in research skill and updating knowledge in pathophysiology should also receive due attention in designing trainings.

National Continuing Professional Development (CPD) training needs of pharmacists in Ethiopia.

BACKGROUND: Continuing Professional Development (CPD) in pharmacy is a lifelong learning approach whereby individual pharmacists are responsible for updating and broadening their knowledge, skills, and attitudes. This is vital to ensure the delivery of high-quality patient care services. However, there is a lack of available data revealing the CPD needs of Ethiopian pharmacists. Thus, the objective of this study was to identify CPD training needs of pharmacists practicing in Ethiopia. METHODS: An institution-based cross-sectional study design with a quantitative approach was employed in this study. This assessment involved 640 pharmacists representing various sectors of the profession. Data were collected through a combination of an online platform and a face-to-face questionnaire administered in person. RESULT: A total of 634 participants completed and returned the questionnaires, resulting in an impressive response rate of 99.1%. A significant majority (74.1%) of the participants possessed bachelor's degree in pharmacy (B. Pharm). Pharmaceutical Logistics and Pharmacy administration was preferentially selected as a prior CPD course by 36% of participants, of them while Pharmacotherapy (17%), Leadership/Governance (13%), Community Pharmacy (12%), Research and Development (11%) were also the subsequent top choices by participants. Off-site face-to-face lectures (59.2%), Hybrid (face-to-face + e-learning) (54.8%), and on-site on-the-job training (45.5%) were the most convenient means of CPD course delivery. On the other hand, the participants least favored print-based or correspondence programs for CPD course delivery. CONCLUSIONS: CPD holds great importance in the professional lives of pharmacists. It is critical for pharmacists, CPD providers, and those responsible for accrediting CPD programs to recognize the specific CPD requirements, preferred methods of delivery, and obstacles involved. This understanding is vital for establishing priorities and effectively planning CPD activities. In light of this, our study identified the most preferred CPD training courses and convenient delivery methods for pharmacists in Ethiopia. We recommend that CPD providers and accrediting bodies in Ethiopia refer to our findings when approving CPD courses.

The 6-4 photoproduct is the trigger of UV-induced replication blockage and ATR activation.

The most prevalent human carcinogen is sunlight-associated ultraviolet (UV), a physiologic dose of which generates thousands of DNA lesions per cell, mostly of two types: cyclobutane pyrimidine dimers (CPDs) and 6-4 photoproducts (6-4PPs). It has not been possible, in living cells, to precisely characterize the respective contributions of these two lesion types to the signals that regulate cell cycle progression, DNA replication, and cell survival. Here we coupled multiparameter flow cytometry with lesion-specific photolyases that eliminate either CPDs or 6-4PPs and determined their respective contributions to DNA damage responses. Strikingly, only 6-4PP lesions activated the ATR-Chk1 DNA damage response pathway. Mechanistically, 6-4PPs, but not CPDs, impeded DNA replication across the genome as revealed by microfluidic-assisted replication track analysis. Furthermore, single-stranded DNA accumulated preferentially at 6-4PPs during DNA replication, indicating selective and prolonged replication blockage at 6-4PPs. These findings suggest that 6-4PPs, although eightfold fewer in number than CPDs, are the trigger for UV-induced DNA damage responses.