THE FUTURE OF NEXT-GENERATION SEQUENCING FOR BLOOD GROUP GENOTYPING AND ITS IMPLICATIONS IN TRANSFUSION MEDICINE

Alloimmunisation becomes a problem when serological discrepancies occur in matching antigens between donors and patients for blood transfusion. The rate of alloimmunisation has been increased especially in multiply transfused patients. Blood group genotyping (BGG) is a DNA-based assay that aids in reducing this situation. Currently, many platforms of BGG have become available, in which every technique has its own advantages and disadvantages. All these platforms lack the ability to identify novel alleles that might have an unknown clinical significance. The advent of next-generation sequencing (NGS) offers identification of the unprecedented alleles due to its basis of sequence-based typing. Moreover, it provides an extreme high-throughput which may be able to screen many donors and patients in a single run. In this project, two approaches have been developed in generating sequencing libraries followed by sequencing on the Ion Torrent Personal Genome MachineTM platform (Ion PGMTM). The first approach was amplicon-based target selection using Ion AmpliseqTM Custom Panel, designated as Human Erythrocyte Antigens and Human Platelet Antigens Panel (HEA and HPA Panel). This panel assay screens 11 blood group systems, as well as 16 human platelet antigens. The outcome was extraordinary, in particular four novel alleles had been identified out of 28 samples, one in the RHCE gene 208C>T (Arg70Trp) in exon 2 and three in the KEL gene. The first SNP was 331G>A (Ala111Thr) in exon 4. The second SNP was 1907C>T (Ala636Val) in exon 17 and the third SNP was 2165T>C (Leu722Pro) in exon 19. However, some issues occurred regarding co-amplification of homologous genes. The second approach was a long-range polymerase chain reaction (LR-PCR) based approach. This method provided a high resolution assay by amplification of entire genes, including the non-coding area, of the Kell and Rh blood group systems. The Kell blood group was initially utilised as a model in order to apply the same approach on the Rh system. Most alleles encoding the antigens of the Kell blood group, especially the high prevalence ones, were identified. The Rh LR-PCR approach was carried out by amplification of the RHD and RHCE genes with seven amplicons. For five RhD-positive samples no mutations were observed within the coding areas. On the other hand, five serotyped weak D samples were genotyped as; two weak D type 1, two weak D type 2 and one DAR3.1 weak partial D 4.0 (RHD*DAR3.01). Regarding the RHCE, the following antigens (C, c, E, c) were predicted properly from the sequencing data. Moreover, the RHCE*ceVS.02 was identified. 64 and 39 intronic SNPs were identified in RHD and RHCE genes, respectively. The intronic SNPs assisted the genotyping process by identifying the haplotype of interest. Interestingly, the novel allele identified in the RHCE gene by the HEA and HPA Panel was confirmed to belong to the RHCE gene by the LR-PCR approach, indicating the panel misaligned it to the RHD gene. In conclusion, NGS paves the way to be an alternative substitution to the previous molecular techniques. It would supplant the conventional serology for typing blood for transfusion.Jazan University, Kingdom of Saudi Arabi

Health professions’ students have an alarming prevalence of depressive symptoms: exploration of the associated factors

Contains fulltext : 172650.pdf (publisher's version ) (Open Access)BACKGROUND: There is a need to better understand the depression phenomenon and to clarify why some students become depressed and others don't. The purpose of this study was to compare the prevalence of depressive symptoms among health professions' (HP) students, and to explore the association between socio-demographic factors (e.g. year of study, discipline, gender) and depressive symptoms. METHODS: In this descriptive-analytic, cross-sectional study, stratified proportionate sampling strategy was used to select the study sample during the academic year 2012-2013. The students from four health professions' schools situated within a large, public university located in Riyadh, Saudi Arabia were screened for depressive symptoms using the 21-item Beck Depression Inventory (BDI II). Chi-square test, student t-test and ANOVA were used to compare different categorical variables. RESULTS: The overall response rate was 79.0 %, the highest among dental students 86.1 %, and lowest among nursing (49.7 %). The overall prevalence rate of depressive symptoms was 47.0 %; it was highest among dentistry students (51.6 %), followed by medicine (46.2 %), applied medical sciences (AMS) (45.7 %) and lowest among nursing students (44.2 %). A statistically significant association was found between the presence and severity of depressive symptoms on one hand and the female gender (p = 0.000) and year of study on the other hand. CONCLUSION: This study seems to indicate an alarming rate of depressive symptoms. Female gender, dentistry, the third year for all schools and fifth year for medicine and dentistry have the highest association with depressive symptoms. Future studies may be needed to explore further the reasons and explanations for the variation in the prevalence of depressive symptoms among these groups. The factors that deserve exploration include curricular variables and personal factors such as the students' study skills

Usefulness of routine pairing of anaerobic with aerobic blood culture bottles and decision making on antimicrobial therapy

ObjectivesTo evaluate the growth concordance in paired aerobic/anaerobic sets, and the impact of the anaerobic growth on patients' antimicrobial management.MethodThis is a prospective multicenter study which was conducted in three hospitals, with total beds of 750 beds and 52 ICU beds. Prospectively, laboratory blood cultures logbooks were daily reviewed and patients from whom blood cultures were ordered were followed, their chart were reviewed. Entries on antimicrobial therapeutic changes were noted for all paired sets. Clinicians were blinded to the study, though they were informed about culture results via the usual work protocol in each hospital.ResultsCollected Blood culture sets totaled 2492; 172 single sets were excluded, and 1160 paired sets were analyzed. 1046 were concordant; 79 sets had bacterial growth and 967 sets had no bacterial growth. 114 sets were discordant; 97 in aerobic bottles, 13 in anaerobic, and 4 in both.The proportion of agreement for the concordant paired growth sets was 90.2%.  The composite proportion of agreement for sets with any growth (N = 193, composite proportion of agreement = 56%, 95% C.I., 34% - 48%). Cohen kappa composite agreement, measured for the total analyzed paired-sets (N = 1160, K = .52, SE = .038. 95% C.I., .447 - .595). The odds of modifying antimicrobial regimen were for total and subgroups intent to treat odds, based on paired sets showed that one modification took place in one anaerobic growth set (N = 1160, Odds = 0.0008), the odds for all sets with any growth (N = 193, odds = .005), and based on any anaerobic sets (79 concordant, 13 anaerobic, and 4 discordant) with bacterial growth (N = 96: odds = 0.010).ConclusionThe study demonstrates that the proportion of agreement among paired sets were high, and needless to include anaerobic sets in routine blood culture collection. Also the decision-making of anti-infective treatment on patients based on anaerobic blood culture growth was not evident

Medical doctors\u27 offline computer-assisted digital education: Systematic review by the digital health education collaboration

Background: The widening gap between innovations in the medical field and the dissemination of such information to doctors may affect the quality of care. Offline computer-based digital education (OCDE) may be a potential solution to overcoming the geographical, financial, and temporal obstacles faced by doctors.Objective: The objectives of this systematic review were to evaluate the effectiveness of OCDE compared with face-to-face learning, no intervention, or other types of digital learning for improving medical doctors\u27 knowledge, cognitive skills, and patient-related outcomes. Secondary objectives were to assess the cost-effectiveness (CE) of OCDE and any adverse effects.Methods: We searched major bibliographic databases from 1990 to August 2017 to identify relevant articles and followed the Cochrane methodology for systematic reviews of intervention.Results: Overall, 27 randomized controlled trials (RCTs), 1 cluster RCT (cRCT), and 1 quasi-RCT were included in this review. The total number of participants was 1690 in addition to the cRCT, which included 24 practices. Due to the heterogeneity of the participants, interventions, and outcomes, meta-analysis was not feasible, and the results were presented as narrative summary. Compared with face-to-face learning, the effect of OCDE on knowledge gain is uncertain (ratio of the means [RM] range 0.95-1.17; 8 studies, 495 participants; very low grade of evidence). From the same comparison, the effect of OCDE on cognitive skill gain is uncertain (RM range 0.1-0.9; 8 studies, 375 participants; very low grade of evidence). OCDE may have little or no effect on patients\u27 outcome compared with face-to-face education (2 studies, 62 participants; low grade of evidence). Compared with no intervention, OCDE may improve knowledge gain (RM range 1.36-0.98; 4 studies, 401 participants; low grade of evidence). From the same comparison, the effect of OCDE on cognitive skill gain is uncertain (RM range 1.1-1.15; 4 trials, 495 participants; very low grade of evidence). One cRCT, involving 24 practices, investigated patients\u27 outcome in this comparison and showed no difference between the 2 groups with low-grade evidence. Compared with text-based learning, the effect of OCDE on cognitive skills gain is uncertain (RM range 0.91-1.46; 3 trials with 4 interventions; 68 participants; very low-grade evidence). No study in this comparison investigated knowledge gain or patients\u27 outcomes. One study assessed the CE and showed that OCDE was cost-effective when compared with face-to-face learning if the cost is less than or equal to Can $200. No trial evaluated the adverse effect of OCDE.Conclusions: The effect of OCDE compared with other methods of education on medical doctors\u27 knowledge and cognitive skill gain is uncertain. OCDE may improve doctors\u27 knowledge compared with no intervention but its effect on doctors\u27 cognitive skills is uncertain. OCDE may have little or no effect in improving patients\u27 outcome

Anxiety and depression following cumulative low-level exposure to organophosphate pesticides

Previous research suggests that individuals with a prior history of pesticide poisoning are at increased risk of psychiatric disorder (Freire and Koifman, 2013), but findings regarding the impact of cumulative low-level exposure are inconsistent. The aim of the current study was to investigate whether sheep farmers with a history of low-level exposure to organophosphate pesticides (1) report a higher level of psychological distress on subjective symptom questionnaires, compared to unexposed controls (2) also meet internationally agreed diagnostic criteria for a psychiatric disorder more often than unexposed controls. 127 sheep farmers were evaluated and compared to 78 unexposed controls, matched in terms of gender, education, level of intelligence, working status and area of residence. Both self-report measures and structured clinical interviews were used to assess mental health. The exposed cohort reported significantly higher rates of anxiety and depression when self-report questionnaires were used to evaluate mood, even when stressful life events, demographic and physical health factors were taken into account. However, when diagnostic interviews were used to assess mood, this pattern only held true for anxiety

Systematic review and meta-analysis of the effectiveness of pre-pregnancy care for women with diabetes for improving maternal and perinatal outcomes.

BACKGROUND: Pre-gestational diabetes mellitus is associated with increased risk of maternal and perinatal adverse outcomes. This systematic review was conducted to evaluate the effectiveness and safety of pre-conception care (PCC) in improving maternal and perinatal outcomes. METHODS: Databases from MEDLINE, EMBASE, WEB OF SCIENCE, and Cochrane Library were searched, including the CENTRAL register of controlled trials, and CINHAL up until March 2019, without any language restrictions, for any pre-pregnancy care aiming at health promotion, glycemic control, and screening and treatment of diabetes complications in women with type I or type II pre-gestational diabetes. Trials and observational studies were included in the review. Newcastle-Ottawa scale and the Cochrane collaboration methodology for data synthesis and analysis were used, along with the GRADE tool to evaluate the body of evidence. RESULTS: The search identified 8500 potentially relevant citations of which 40 reports of 36 studies were included. The meta-analysis results show that PCC reduced congenital malformations risk by 71%, (Risk ratio (RR) 0.29; 95% CI: 0.21-0.40, 25 studies; 5903 women; high-certainty evidence). The results also show that PCC may lower HbA1c in the first trimester of pregnancy by an average of 1.27% (Mean difference (MD) 1.27; 95% CI: 1.33-1.22; 4927 women; 24 studies, moderate-certainty evidence). Furthermore, the results suggest that PCC may lead to a slight reduction in the risk of preterm delivery of 15%, (RR 0.85; 95% CI: 0.73-0.99; nine studies, 2414 women; moderate-certainty evidence). Moreover, PCC may result in risk reduction of perinatal mortality by 54%, (RR 0.46; 95% CI: 0.30-0.73; ten studies; 3071 women; moderate-certainty evidence). There is uncertainty about the effects of PCC on the early booking for antenatal care (MD 1.31; 95% CI: 1.40-1.23; five studies, 1081 women; very low-certainty evidence) and maternal hypoglycemia in the first trimester, (RR 1.38; 95% CI: 1.07-1.79; three studies; 686 women; very low- certainty evidence). In addition, results of the meta-analysis indicate that PCC may lead to 48% reduction in the risk of small for gestational age (SGA) (RR 0.52; 95% CI: 0.37-0.75; six studies, 2261 women; moderate-certainty evidence). PCC may reduce the risk of neonatal admission to intensive care unit (NICU) by 25% (RR 0.75; 95% CI: 0.67-0.84; four studies; 1322 women; moderate-certainty evidence). However, PCC may have little or no effect in reducing the cesarean section rate (RR 1.02; 95% CI: 0.96-1.07; 14 studies; 3641 women; low-certainty evidence); miscarriage rate (RR 0.86; 95% CI: 0.70-1.06; 11 studies; 2698 women; low-certainty evidence); macrosomia rate (RR 1.06; 95% CI: 0.97-1.15; nine studies; 2787 women, low-certainty evidence); neonatal hypoglycemia (RR 0.93; 95% CI: 0.74-1.18; five studies; 880 women; low-certainty evidence); respiratory distress syndrome (RR 0.78; 95% CI: 0.47-1.29; four studies; 466 women; very low-certainty evidence); or shoulder dystocia (RR 0.28; 95% CI: 0.07-1.12; 2 studies; 530 women; very low-certainty evidence). CONCLUSION: PCC for women with pre-gestational type 1 or type 2 diabetes mellitus is effective in improving rates of congenital malformations. In addition, it may improve the risk of preterm delivery and admission to NICU. PCC probably reduces maternal HbA1C in the first trimester of pregnancy, perinatal mortality and SGA. There is uncertainty regarding the effects of PCC on early booking for antenatal care or maternal hypoglycemia during the first trimester of pregnancy. PCC has little or no effect on other maternal and perinatal outcomes

Virtual Handover of Patients in the Pediatric Intensive Care Unit During the Covid-19 Crisis

Objective: A key measure to mitigate coronavirus disease 2019 (COVID-19) has been social distancing. Incorporating video-conferencing applications in the patient handover process between healthcare workers can enhance social distancing while maintaining handover elements. This study describes pediatric intensive care unit (PICU) physicians' experience using an online video-conferencing application for handover during the COVID-19 pandemic. Design: Qualitative content analysis. Setting: PICU at a university hospital in Riyadh, Saudi Arabia. Subjects: PICU Physicians. Interventions: Due to the pandemic, the hospital's PICU used Zoom® as a remote conferencing application instead of a face-to-face handover. Following institutional review board approval, data were collected over two weeks (1 Jul 2020 to 14 Jul 2020). Measurements: An online survey was conducted using open-ended questions to capture demographic data and the perceived efficacy of remote handovers. Thematic framework analysis process included open coding, creating categories, and abstraction. Main Results: All 37 PICU physicians who participated in the handover completed the survey. The participants comprised six attendings, nine specialists, and 22 residents. While 20 (54.1%) physicians reported attending 1-5 Zoom handovers by the time of the study, some (n. 6, 16.2%) had more than ten virtual handovers. They had variable previous teleconferencing experiences. Most physicians (78.4%) were comfortable conducting a remote handover. Most found that Situation-Background-Assessment-Recommendation handover elements were properly achieved through this remote handover process. The perceived advantages of online handover included fewer interruptions, time efficiency, and facilitation of social distancing. The perceived disadvantages were the paucity of nonverbal communication and teaching during virtual meetings. Conclusion: Video-conferencing applications for online handovers could supplement traditional face-to-face intensive care unit patient handover during outbreaks of infectious diseases. The use of video streaming and more emphasis on teaching should be encouraged to optimize the users' experience

Sustained proliferation in cancer: mechanisms and novel therapeutic targets

Proliferation is an important part of cancer development and progression. This is manifest by altered expression and/or activity of cell cycle related proteins. Constitutive activation of many signal transduction pathways also stimulates cell growth. Early steps in tumor development are associated with a fibrogenic response and the development of a hypoxic environment which favors the survival and proliferation of cancer stem cells. Part of the survival strategy of cancer stem cells may manifested by alterations in cell metabolism. Once tumors appear, growth and metastasis may be supported by overproduction of appropriate hormones (in hormonally dependent cancers), by promoting angiogenesis, by undergoing epithelial to mesenchymal transition, by triggering autophagy, and by taking cues from surrounding stromal cells. A number of natural compounds (e.g., curcumin, resveratrol, indole-3-carbinol, brassinin, sulforaphane, epigallocatechin-3-gallate, genistein, ellagitannins, lycopene and quercetin) have been found to inhibit one or more pathways that contribute to proliferation (e.g., hypoxia inducible factor 1, nuclear factor kappa B, phosphoinositide 3 kinase/Akt, insulin-like growth factor receptor 1, Wnt, cell cycle associated proteins, as well as androgen and estrogen receptor signaling). These data, in combination with bioinformatics analyses, will be very important for identifying signaling pathways and molecular targets that may provide early diagnostic markers and/or critical targets for the development of new drugs or drug combinations that block tumor formation and progression

Adenovirus and RNA-based COVID-19 vaccines’ perceptions and acceptance among healthcare workers in Saudi Arabia: a national survey

Objectives The aim of this study was to compare the perception, confidence, hesitancy and acceptance rate of various COVID-19 vaccine types among healthcare workers (HCWs) in Saudi Arabia, a nation with Middle East respiratory syndrome coronavirus experience. Design National cross-sectional, pilot-validated questionnaire. Setting Online, self-administered questionnaire among HCWs. Participants A total of 2007 HCWs working in the Kingdom of Saudi Arabia participated; 1512 (75.3%) participants completed the survey and were included in the analysis. Intervention Data were collected through an online survey sent to HCWs during 1–15 November 2020. The main outcome measure was HCW acceptance of COVID-19 candidate vaccines. The associated factors of vaccination acceptance were identified through a logistic regression analysis and via measurement of the level of anxiety, using the Generalised Anxiety Disorder 7 scale. Results Among the 1512 HCWs who were included, 62.4% were women, 70.3% were between 21 and 40 years of age, and the majority (62.2%) were from tertiary hospitals. In addition, 59.5% reported knowing about at least one vaccine; 24.4% of the participants were sure about their willingness to receive the ChAdOx1 nCoV-19 vaccine, and 20.9% were willing to receive the RNA BNT162b2 vaccine. However, 18.3% reported that they would refuse to receive the Ad5-vectored vaccine, and 17.9% would refuse the Gam-COVID-Vac vaccine. Factors that influenced the differential readiness of HCWs included their perceptions of the vaccine’s efficiency in preventing the infection (33%), their personal preferences (29%) and the vaccine’s manufacturing country (28.6%). Conclusions Awareness by HCWs of the several COVID-19 candidate vaccines could improve their perceptions and acceptance of vaccination. Reliable sources on vaccine efficiency could improve vaccine uptake, so healthcare authorities should use reliable information to decrease vaccine hesitancy among frontline healthcare providers

SARS-CoV-2 B.1.1.7 UK Variant of Concern Lineage-Related Perceptions, COVID-19 Vaccine Acceptance and Travel Worry Among Healthcare Workers

Background: Healthcare workers' (HCWs') travel-related anxiety needs to be assessed in light of the emergence of SARS-CoV-2 mutations. Methods: An online, cross-sectional questionnaire among HCWs between December 21, 2020 to January 7, 2021. The outcome variables were HCWs' knowledge and awareness of the SARS-CoV-2 B.1.1.7 lineage that was recently reported as the UK variant of concern, and its associated travel worry and Generalized Anxiety Disorder (GAD-7) score. Results: A total of 1,058 HCWs completed the survey; 66.5% were female, 59.0% were nurses. 9.0% indicated they had been previously diagnosed with COVID-19. Regarding the B.1.1.7 lineage, almost all (97.3%) were aware of its emergence, 73.8% were aware that it is more infectious, 78.0% thought it causes more severe disease, and only 50.0% knew that current COVID-19 vaccines are effective in preventing it. Despite this, 66.7% of HCWs were not registered to receive the vaccine. HCWs' most common source of information about the new variant was social media platforms (67.0%), and this subgroup was significantly more worried about traveling. Nurses were more worried than physicians (P = 0.001). Conclusions: Most HCWs were aware of the emergence of the SARS-CoV-2 B.1.1.7 variant and expressed substantial travel worries. Increased worry levels were found among HCWs who used social media as their main source of information, those with lower levels of COVID-19 vaccine uptake, and those with higher GAD-7 scores. The utilization of official social media platforms could improve accurate information dissemination among HCWs regarding the Pandemic's evolving mutations. Targeted vaccine campaigns are warranted to assure HCWs about the efficacy of COVID-19 vaccines toward SARS-CoV-2 variants