The need for and acceptability of a curriculum to train nursing and medical students in the sexual healthcare of clients with female genital mutilation/cutting in Tanzania.

BACKGROUND: Female genital mutilation/cutting (FGM/C) is tied to one of the most conservative cultures in the Mediterranean and Sub-Saharan Africa. More than 200 million girls and women in 30 African, Asian and the middle Eastern countries have undergone FGM/C. However, healthcare professionals are not adequately trained to prevent and manage FGM/C-related complications including sexual health problems. This study aimed to assess the need and acceptability of a curriculum to train nursing and medical students in the sexual healthcare of clients with FGM/C in Tanzania. METHODS: We used a descriptive and cross sectional study design to collect and analyse information from 271 medical and 137 nursing students in Tanzania. A Qualtrics online survey was used to obtain quantitative data on training interest, previous training received, and the curriculum delivery method. Open-ended questions were used to explore their insights on significance to obtain the necessary competencies to treat and prevent FGM/C. Descriptive statistics were used to analyze quantitative data while qualitative data were analyzed using a thematic approach. RESULTS: Almost half of the participants reported they had little to no training in sexual healthcare for women with FGM/C (47%). In all, 82.4% reported the training to be acceptable. Following thematic analysis of open-ended questions, participants expressed a desire to improve their competencies to meet the current and future sexual and psychological health needs of women and girls who have undergone FGM/C. CONCLUSION: It is a necessary and acceptable to develop a curriculum to train healthcare students to diagnose, treat and prevent sexual health complications related to FGM/C. In our study, designing a culturally sensitive curriculum and its delivery method, that includes practical sessions with simulated patients, was considered the most beneficial and favorable.

Addressing sexual health concerns in Tanzania: perceived barriers among healthcare professionals and students in the "training for health professionals" study.

Little is known about the factors that may prevent healthcare professionals as key stakeholders from exploring sexual health issues in Tanzania. This study examined healthcare professionals' perspectives on the barriers to addressing sexual health concerns in practice. In June 2019, we conducted an exploratory qualitative study involving 18 focus group discussions among healthcare professionals (n = 60) and students (n = 61) in the health professions (midwifery, nursing, medicine) in Dar es Salaam, Tanzania. Study participants and design were purposively selected and stratified. We used a focus group discussion guide in Kiswahili. Data were transcribed in Kiswahili and translated into English. A thematic analysis approach was used for data analysis. Two themes were developed: (1) differences between health care professional and patient socio-demographic characteristics; (2) health care system and patients' backgrounds, such as communication barriers, lack of confidentiality and privacy within health facilities, type of clinical presentation and complaint, patient behaviours, and their clinical background. Several key barriers prevented sexual health communication between healthcare professionals and patients, affecting the quality of sexual health service delivery. Additional sexual health clinical training is warranted for health professions students and professionals to optimise sexual health care delivery in a culturally conservative country like Tanzania.

Healthcare professional students' skills in sexual health communication and history taking: inter-rater reliability of standardized patients and faculty ratings in dar es Salaam, tanzania- a cross-sectional study.

BACKGROUND: Low- and middle-income countries face a disproportionate impact of sexual health problems compared to high-income countries. To address this situation proper interpersonal communication skills are essential for clinician to gather necessary information during medical history-taking related to sexual health. This study aimed to evaluate the interrater reliability of ratings on sexual health-related interpersonal communication and medical history-taking between SPs and trained HCP faculty for health care professional students. METHODS: We conducted a cross-sectional comparative study to evaluate the interrater reliability of ratings for sexual health-related interpersonal communication and medical history-taking. The data were collected from medical and nursing students at Muhimbili University of Health and Allied Sciences, who interviewed 12 Standardized Patients (SPs) presenting with sexual health issues. The video-recorded interviews rated by SPs, were compared to the one rated by 5 trained Health Care Professional (HCP) faculty members. Inter-rater reliability was evaluated using percent agreement (PA) and kappa statistics (κ). RESULTS: A total of 412 students (mean age 24) were enrolled in the study to conduct interviews with two SPs presenting with sexual health concerns. For interpersonal communication (IC), the overall median agreement between raters was slight (κ2 0.0095; PA 48.9%) while the overall median agreement for medical sexual history-taking was deemed fair (κ2 0.139; PA 75.02%). CONCLUSION: The use of SPs for training and evaluating medical and nursing students in Tanzania is feasible only if they undergo proper training and have sufficient time for practice sessions, along with providing feedback to the students.

The need for and acceptability of a cancer training course for medical and nursing students in Tanzania: a convergent mixed methods study.

BACKGROUND: The cancer burden in Africa is on the rise. A Cancer Training Course on screening, prevention, care, and community education is crucial for addressing a wide range of cancer health issues. When appropriately educated healthcare providers on cancer provide care, patient care improves, and healthcare costs decrease. However, in Tanzania, doctors and nurses receive little or no training in primary cancer care in their bachelor's program. AIM: This study assessed the need and acceptability of a cancer training course for nursing and medical doctor students at the Muhimbili University of Health and Allied Sciences (MUHAS) in Dar es Salaam, Tanzania. METHODS: This study was a cross-sectional parallel mixed method study during the 3-month follow-up within the larger study on sexual health training for health professionals. The study was a randomized controlled (RCT), single-blind, parallel trial of sexual health training versus a waitlist control among health students at MUHAS in Tanzania. Descriptive analysis was performed to analyze the participants' demographic information, need, and acceptability of the cancer training courseto determine the frequencies and percentages of their distribution between disciplines. In addition, inductive thematic analysis was performed for the qualitative data. The RCT study was registered at Clinical Trial.gov (NCT03923582; 01/05/2021). RESULTS: Data were collected from 408 students (272 medical doctors and 136 nursing students). The median age of the participants was 23 years. Most (86.0%) medical and 78.1% of nursing students reported receiving little to no cancer training. On the other hand, most (92.3%) medical and nursing (92.0%) students were interested in receiving cancer training. Furthermore, 94.1% of medical and 92.0% of nursing students needed a cancer training course in their undergraduate program. In addition, participants said a cancer training course would be important because it would help them improve the quality of cancer care and enhance the quality of life for patients by ensuring early diagnosis and treatment. CONCLUSION: A cancer training course is both highly needed and acceptable to medical and nursing students. Implementation of this cancer training course will improve students' knowledge and skills and eventually improve the quality of cancer care and patients' quality of life by ensuring early diagnosis and management.

MiR-128-3p as blood based liquid biopsy biomarker in childhood acute lymphoblastic leukemia.

BACKGROUND: Minimal residual disease (MRD) is one of the most valuable independent prognostic factors in acute lymphoblastic leukemia (ALL). Bone marrow (BM) aspiration, however, is an invasive process. Previous studies have shown that microRNAs (miR) and extracellular vesicle (EV)-related miRs show different expression profiles at the presence of malignant cells compared to healthy controls. In our previous project, we have reported that two miRs previously described to be overexpressed in blasts were significantly decreased over the first week of the therapy of patients with ALL in the platelet free plasma fraction (PFP) of peripheral blood samples (PB). The aim of the current study was to assess the relation between day 15 flow cytometry (FC) MRD and expression of miR-128-3p and miR-222-3p miRs in exosome-enriched fraction (EEF) of PFP to evaluate whether their expression in EEF correlates with day 15 FC MRD more precisely. METHODS: PB was collected from 13 patients diagnosed with pediatric pre-B ALL at 4 time points. Expression of miR-128-3p and miR-222-3p was measured by qPCR in PFP and EEF. RESULTS: Positive correlation was found between changes of miR-128-3p expression in EEF or PFP by day 8 of chemotherapy and day 15 FC MRD (rEEF = 0.99, pEEF = 1.13E-9 and rPFP = 0.99, pPFP = 4.75E-9, respectively). Furthermore, the decrease of miR-128-3p in EEF by day 15 of treatment also showed a positive correlation with day 15 FC MRD (rEEF = 0.96; pEEF = 4.89E-5). CONCLUSION: Our results show that circulating miRs are potential biomarkers of ALL MRD, asmiR-128-3p level both in PFP and EEF predicts day 15 FC MRD. In addition, the assessment of the EEF gave a more promising result.

Structure, function, and evolution of Gga-AvBD11, the archetype of the structural avian-double-ß-defensin family.

Out of the 14 avian ß-defensins identified in the Gallus gallus genome, only 3 are present in the chicken egg, including the egg-specific avian ß-defensin 11 (Gga-AvBD11). Given its specific localization and its established antibacterial activity, Gga-AvBD11 appears to play a protective role in embryonic development. Gga-AvBD11 is an atypical double-sized defensin, predicted to possess 2 motifs related to ß-defensins and 6 disulfide bridges. The 3-dimensional NMR structure of the purified Gga-AvBD11 is a compact fold composed of 2 packed ß-defensin domains. This fold is the archetype of a structural family, dubbed herein as avian-double-ß-defensins (Av-DBD). We speculate that AvBD11 emanated from a monodomain gene ancestor and that similar events might have occurred in arthropods, leading to another structural family of less compact DBDs. We show that Gga-AvBD11 displays antimicrobial activities against gram-positive and gram-negative bacterial pathogens, the avian protozoan Eimeria tenella, and avian influenza virus. Gga-AvBD11 also shows cytotoxic and antiinvasive activities, suggesting that it may not only be involved in innate protection of the chicken embryo, but also in the (re)modeling of embryonic tissues. Finally, the contribution of either of the 2 Gga-AvBD11 domains to these biological activities was assessed, using chemically synthesized peptides. Our results point to a critical importance of the cationic N-terminal domain in mediating antibacterial, antiparasitic, and antiinvasive activities, with the C-terminal domain potentiating the 2 latter activities. Strikingly, antiviral activity in infected chicken cells, accompanied by marked cytotoxicity, requires the full-length protein.

Fear of childbirth: validation of the Kiswahili version of Wijma delivery expectancy/experience questionnaire versions A and B in Tanzania.

BACKGROUND: Fear of childbirth is common both before and after childbirth, often leading to complications in mother and new-born. The Wijma Delivery Expectancy/Experience Questionnaires (W-DEQ) are commonly used to measure fear of childbirth among women before (version A) and after childbirth (version B). The tools are not yet validated in the Tanzanian context. This study aimed to validate the reliability, validity, and factorial structure of their Kiswahili translations. METHODS: A longitudinal study was conducted in six public health facilities in the Pwani region, Tanzania. In all, 694 pregnant and 625 postnatal women were concurrently selected and responded to W-DEQ-A and W-DEQ-B. Validation involved: translating the English questionnaires into Kiswahili; expert rating of the relevancy of the Kiswahili versions' items; computing content validity ratio; piloting the tools; data collection; statistical analysis with reliability evaluated using Cronbach's alpha and the intraclass correlation coefficient. Tool validity was assessed using factor analysis, convergent and discriminant validity. Exploratory factor analysis and confirmatory factor analysis were conducted on data collected using W-DEQ-A and W-DEQ-B, respectively. RESULTS: Exploratory factor analysis revealed seven factors contributing to 50% of the total variation. Four items did not load to any factor and were deleted. The factors identified were: fear; lack of self-efficacy; lack of positive anticipation; isolation; concerns for the baby; negative emotions; lack of positive behaviour. The factors correlated differently with each other and with the total scores. Both Kiswahili versions with 33 items had good internal consistency, with Cronbach's alphas of .83 and .85, respectively. The concerns for the baby factor showed both convergent and discriminant validity. The other six factors showed some problems with convergent validity. The final model from the confirmatory factor analysis yielded 29 items with good psychometric properties (χ2/df = 2.26, p = < .001, RMSEA = .045, CFI = .90 and TLI = .81). CONCLUSIONS: The Kiswahili W-DEQ-A-Revised and W-DEQ-B-Revised are reliable tools and measure fear of childbirth with a multifactorial structure, encompassing seven factors with 29 items. They are recommended for measuring fear of childbirth among pregnant and postnatal Tanzanian women. Further studies are needed to address the inconsistent convergent validity in the revised versions and assess the psychometric properties of W-DEQ-A among pregnant women across gestational ages.

Functional Diversification of Oyster Big Defensins Generates Antimicrobial Specificity and Synergy against Members of the Microbiota.

Big defensins are two-domain antimicrobial peptides (AMPs) that have highly diversified in mollusks. Cg-BigDefs are expressed by immune cells in the oyster Crassostrea gigas, and their expression is dampened during the Pacific Oyster Mortality Syndrome (POMS), which evolves toward fatal bacteremia. We evaluated whether Cg-BigDefs contribute to the control of oyster-associated microbial communities. Two Cg-BigDefs that are representative of molecular diversity within the peptide family, namely Cg-BigDef1 and Cg-BigDef5, were characterized by gene cloning and synthesized by solid-phase peptide synthesis and native chemical ligation. Synthetic peptides were tested for antibacterial activity against a collection of culturable bacteria belonging to the oyster microbiota, characterized by 16S sequencing and MALDI Biotyping. We first tested the potential of Cg-BigDefs to control the oyster microbiota by injecting synthetic Cg-BigDef1 into oyster tissues and analyzing microbiota dynamics over 24 h by 16S metabarcoding. Cg-BigDef1 induced a significant shift in oyster microbiota ß-diversity after 6 h and 24 h, prompting us to investigate antimicrobial activities in vitro against members of the oyster microbiota. Both Cg-BigDef1 and Cg-BigDef5 were active at a high salt concentration (400 mM NaCl) and showed broad spectra of activity against bacteria associated with C. gigas pathologies. Antimicrobial specificity was observed for both molecules at an intra- and inter-genera level. Remarkably, antimicrobial spectra of Cg-BigDef1 and Cg-BigDef5 were complementary, and peptides acted synergistically. Overall, we found that primary sequence diversification of Cg-BigDefs has generated specificity and synergy and extended the spectrum of activity of this peptide family.

Real-Time Fluorescence Microscopy on Living E. coli Sheds New Light on the Antibacterial Effects of the King Penguin ß-Defensin AvBD103b.

(1) Antimicrobial peptides (AMPs) are a promising alternative to conventional antibiotics. Among AMPs, the disulfide-rich ß-defensin AvBD103b, whose antibacterial activities are not inhibited by salts contrary to most other ß-defensins, is particularly appealing. Information about the mechanisms of action is mandatory for the development and approval of new drugs. However, data for non-membrane-disruptive AMPs such as ß-defensins are scarce, thus they still remain poorly understood. (2) We used single-cell fluorescence imaging to monitor the effects of a ß-defensin (namely AvBD103b) in real time, on living E. coli, and at the physiological concentration of salts. (3) We obtained key parameters to dissect the mechanism of action. The cascade of events, inferred from our precise timing of membrane permeabilization effects, associated with the timing of bacterial growth arrest, differs significantly from the other antimicrobial compounds that we previously studied in the same physiological conditions. Moreover, the AvBD103b mechanism does not involve significant stereo-selective interaction with any chiral partner, at any step of the process. (4) The results are consistent with the suggestion that after penetrating the outer membrane and the cytoplasmic membrane, AvBD103b interacts non-specifically with a variety of polyanionic targets, leading indirectly to cell death.

Aphid BCR4 Structure and Activity Uncover a New Defensin Peptide Superfamily.

Aphids (Hemiptera: Aphidoidea) are among the most detrimental insects for agricultural plants, and their management is a great challenge in agronomical research. A new class of proteins, called Bacteriocyte-specific Cysteine-Rich (BCR) peptides, provides an alternative to chemical insecticides for pest control. BCRs were initially identified in the pea aphid Acyrthosiphon pisum. They are small disulfide bond-rich proteins expressed exclusively in aphid bacteriocytes, the insect cells that host intracellular symbiotic bacteria. Here, we show that one of the A. pisum BCRs, BCR4, displays prominent insecticidal activity against the pea aphid, impairing insect survival and nymphal growth, providing evidence for its potential use as a new biopesticide. Our comparative genomics and phylogenetic analyses indicate that BCRs are restricted to the aphid lineage. The 3D structure of BCR4 reveals that this peptide belongs to an as-yet-unknown structural class of peptides and defines a new superfamily of defensins.

The Management of Masturbation as a Sexual Health Issue in Dar es Salaam, Tanzania: A Qualitative Study of Health Professionals' and Medical Students' Perspectives.

BACKGROUND: Across Africa, there are strong cultural taboos against masturbation. AIM: As part of a broader study investigating sexual health training needs of the health providers, researchers conducted a study to investigate how masturbation is addressed as a clinical issue in clinics in Dar es Salaam, Tanzania. METHODS: An exploratory qualitative study design conducted in June 2019 involving 18 focus groups among health care providers and students in the health professions (midwives, nurses, medical doctors). A total of 61 health care students and 58 health providers were interviewed. The study participants were purposively selected and the design was purposively stratified to examine findings across the 3 main health care providers and by experience (clinicians vs students). A semistructured interview guide in Kiswahili language was used. The study participants were presented a case scenario of a 14-year-old boy who was found masturbating in his room by his father, and asked how this case would be handled in a clinical setting. Data were transcribed in Kiswahili and Translated to English. OUTCOMES: Inductive-deductive thematic analysis was performed. Major themes and subthemes were identified. RESULTS: Two main themes emerged: (i) knowledge about the management of masturbation and (ii) views about the effects of masturbation. Clinical interventions providers would try to include normalization of masturbation as a pubescent behavior combined with advice to stop the adolescent from masturbating, a recommendation to watch for negative effects immediately postmasturbation, and referral to a psychologist for treatment. Across providers and students, masturbation in adolescence was seen as clinically problematic, potentially leading to multiple issues in adulthood including sexual dissatisfaction with a spouse, psychological dependency, and erectile dysfunction, loss of sexual sensitivity in intercourse, premature ejaculation, and penis size reduction. Several participants mentioned they received no training about masturbation to guide their clinical practice. CLINICAL IMPLICATIONS: These findings affirm the need for comprehensive sexual health training in Tanzanian universities. STRENGTHS AND LIMITATIONS: Use of stratified design by profession and experience allowed to explore if there appear to be differences between students and experienced providers. The findings cannot be generalizable to all health professional students and providers across Tanzania. CONCLUSION: When designing sexual health curricula for Tanzania, it is important to include accurate information about masturbation as a normal and healthy sexual practice to address widely held myths about its effects on health, and to train providers in how to counsel when concerns and inaccurate information are brought to the clinical encounter. Mushy SE, Rosser BRS, Ross MW, et al. The Management of Masturbation as a Sexual Health Issue in Dar es Salaam, Tanzania: A Qualitative Study of Health Professionals' and Medical Students' Perspectives. J Sex Med 2021;18:1690-1697.

Cultural and clinical challenges in sexual health care provision to men who have sex with men in Tanzania: a qualitative study of health professionals' experiences and health students' perspectives.

BACKGROUND: Health care providers across sub-Saharan Africa continue to face challenges while delivering sexual health care services. We explored the experiences, views and challenges of health care professionals and health students across different disciplines in Tanzania, towards delivery of sexual health services to men who have sex with men. METHODS: Utilizing a qualitative approach, we recruited 121 health care professionals (providers) and students from the fields of midwifery, nursing and medicine in Dar es Salaam, Tanzania. We conducted 18 focus groups discussions, stratified by profession and experience, to investigate clinical management and challenges while addressing a case of an adult male presenting with rectal gonorrhea. RESULTS: Findings indicated this case as extremely sensitive, clinical management involved establishing rapport and consent, medical care from history taking to treatment, and referral to other specialties. However, the illegal status of homosexuality in Tanzania was a primary concern to participants, this triggered the clinical care of this case scenario as challenging. There were uncertainties whether or not that such a case should be reported to the authorities. CONCLUSION: Findings from this study revealed a need for training health students in Tanzania to address sexual health issues including accurate information on homosexuality, reporting requirements and clinical management in the legal and socio-cultural context of the African continent.

Would you offer contraception to a 14-year-old girl? Perspectives of health students and professionals in Dar es Salaam, Tanzania.

BACKGROUND: Rates of unplanned adolescent pregnancy and unsafe induced abortions are very high in Sub-Saharan African countries including Tanzania. Despite their availability and accessibility, modern family planning methods are reported to be critically underutilized by adolescents. This study is part of a broader study that aims to develop a curriculum that will be used in training health professionals by investigating the sexual health training needs of health providers and students in Tanzania. AIM: This study describes the perceptions of health professionals and students on the provision of contraceptives to adolescents. METHODS: Qualitative formative assessment type of research was conducted using 18 focus groups stratified among health professionals and students (midwives, nurses, and medical doctors). Study participants were presented with the theoretical scenario of a 14-year-old girl who sought contraceptive services at a family planning clinic. This theoretical scenario was used to determine how health professionals and students would handle the case. Thematic analysis guided the examination and determination of data results. RESULTS: Three main themes emerged from the data, including (1) knowledge about the provision of contraceptives to adolescents, (2) perception of the adolescents' right to contraceptive use, and (3) barriers to the provision of contraceptives to adolescents. Participants stated that having a baseline knowledge of contraceptive services for adolescents and their rights to contraceptives would trigger their decision on offering the contraceptive. On the other hand, being unaware of the reproductive health rights for adolescents, judgmental behavior of providers, as well as religious and cultural dynamics were all found to be major barriers for providers to offer contraceptive services to the 14-year-old adolescent girl in the theoretical scenario. CONCLUSION: These findings support the need for comprehensive sexual health education in Tanzanian health professional training programs.

In Tanzania, adolescent pregnancies are culturally unwanted, abortion remains illegal and yet the utilization of family planning services among adolescents is still low. The community myths and misconceptions about the side effects of contraceptives, healthcare providers' attitudes, and barriers within the healthcare system hinder adolescents' utilization of family planning services. The effective use of family planning methods will prevent the use of unsafe induced abortions and hence reduce morbidity and mortality among young women. This will be possible only if healthcare providers are equipped with the knowledge and skills to address these sexual and reproductive health challenges among adolescents in Tanzania. Therefore, in this study, we decided to explore the perceptions of health professionals and students on the provision of contraception to adolescents. We conducted 18 focus groups stratified among health professionals and students (midwives, nurses, medical doctors). Participants were supplied with a theoretical scenario wherein a 14-year-old girl presented to a family planning clinic seeking contraceptive services to determine how health professionals and students would handle the case. We found that the decision of providers with regards to offering contraception is highly dependent on having a baseline knowledge of the rights and contraceptive services available to adolescents. Some barriers identified as reasons for providers not to offer contraceptive services include a lack of awareness of the reproductive health rights for adolescents, judgmental behavior (implicit and explicit biases among providers), as well as religious and cultural dynamics. Participants also acknowledged that the presence of guidelines and proper training among providers might avoid the infringement of adolescents' rights to sexual and reproductive health services. In conclusion, these findings support the need for comprehensive sexual health education in Tanzanian health professional training programs.

Investigation of circulating lncRNAs as potential biomarkers in chronic respiratory diseases.

BACKGROUND: In the present study the blood expression level of inflammatory response and autoimmunity associated long non-coding RNAs (lncRNAs) were compared in patients with different chronic respiratory diseases and investigated whether they could be used as biomarkers in these diseases. METHODS: In the discovery cohort, the gene expression level of 84 lncRNAs were measured in the blood of 24 adult patients including healthy controls and patients with asthma and COPD. In the replication cohort the expression of 6 selected lncRNAs were measured in 163 subjects including healthy controls and adults with allergic rhinitis, asthma, COPD and children with asthma. It was evaluated whether these lncRNAs can be used as diagnostic biomarkers for any studied disease. With systems biology analysis the biological functions of the selected lncRNAs were predicted. RESULTS: In the discovery cohort, the mean expression of 27 lncRNAs showed nominally significant differences in at least one comparison. OIP5-AS1, HNRNPU, RP11-325K4.3, JPX, RP11-282O18.3, MZF1-AS1 were selected for measurement in the replication cohort. Three lncRNAs (HNRNPU, RP11-325K4.3, JPX) expressed significantly higher in healthy children than in adult controls. All the mean expression level of the 6 lncRNAs differed significantly between adult allergic rhinitis patients and controls. RP11-325K4.3, HNRNPU and OIP5-AS1 expressed higher in allergic asthma than in non-allergic asthma. COPD and asthma differed in the expression of RP11-325K4.3 from each other. In examining of the lncRNAs as biomarkers the weighted accuracy (WA) values were especially high in the comparison of healthy controls and patients with allergic rhinitis. OIP5-AS1 and JPX achieved 0.98 and 0.9 WA values, respectively, and the combination of the selected lncRNAs also resulted in a high performance (WA = 0.98). Altogether, OIP5-AS1 had the highest discriminative power in case of three out of six comparisons. CONCLUSION: Differences were detected in the expression of circulating lncRNAs in chronic respiratory diseases. Some of these differences might be utilized as biomarkers and also suggest a possible role of these lncRNAs in the pathomechanism of these diseases. The lncRNAs and the associated pathways are potential therapeutic targets in these diseases, but naturally additional studies are needed for the confirmation of these results.

MicroRNA-181a as novel liquid biopsy marker of central nervous system involvement in pediatric acute lymphoblastic leukemia.

BACKGROUND: Refractory central nervous system (CNS) involvement is among the major causes of therapy failure in childhood acute leukemia. Applying contemporary diagnostic methods, CNS disease is often underdiagnosed. To explore more sensitive and less invasive CNS status indicators, we examined microRNA (miR) expressions and extracellular vesicle (EV) characteristics. METHODS: In an acute lymphoblastic leukemia (ALL) discovery cohort, 47 miRs were screened using Custom TaqMan Advanced Low-Density Array gene expression cards. As a validation step, a candidate miR family was further scrutinized with TaqMan Advanced miRNA Assays on serial cerebrospinal fluid (CSF), bone marrow (BM) and peripheral blood samples with different acute leukemia subtypes. Furthermore, small EV-rich fractions were isolated from CSF and the samples were processed for immunoelectron microscopy with anti-CD63 and anti-CD81 antibodies, simultaneously. RESULTS: Regarding the discovery study, principal component analysis identified the role of miR-181-family (miR-181a-5p, miR-181b-5p, miR-181c-5p) in clustering CNS-positive (CNS+) and CNS-negative (CNS‒) CSF samples. We were able to validate miR-181a expression differences: it was about 52 times higher in CSF samples of CNS+ ALL patients compared to CNS‒ cases (n = 8 vs. n = 10, ΔFC = 52.30, p = 1.5E-4), and CNS+ precursor B cell subgroup also had ninefold higher miR-181a levels in their BM (p = 0.04). The sensitivity of CSF miR-181a measurement in ALL highly exceeded those of conventional cytospin in the initial diagnosis of CNS leukemia (90% vs. 54.5%). Pellet resulting from ultracentrifugation of CNS+ CSF samples of ALL patients showed atypical CD63-/CD81- small EVs in high density by immunoelectron microscopy. CONCLUSIONS: After validating in extensive cohorts, quantification of miR-181a or a specific EV subtype might provide novel tools to monitor CNS disease course and further adjust CNS-directed therapy in pediatric ALL.

Circulating microRNAs as minimal residual disease biomarkers in childhood acute lymphoblastic leukemia.

BACKGROUND: Treatment stratification based on bone marrow minimal residual disease (MRD) at set time points has resulted in considerably improved survival in pediatric acute lymphoblastic leukemia (ALL). Treatment response is assessed using bone marrow samples. MicroRNAs (miRs) easily traffic among fluid spaces and are more stable than most other RNA classes. We examined the role of circulating miRs as putative less invasive MRD biomarkers. METHODS: In an exploratory experiment, expression of 46 preselected miRs was studied in platelet-free blood plasma samples of 15 de novo, 5 relapsed ALL patients and 10 controls by Custom TaqMan Array Advanced MicroRNA Card. Based on their high expression in ALL compared to controls, and on the reduction observed along the induction therapy, four miRs were selected for further analyses: miR-128-3p, -181a-5p, -181b-5p and 222-3p. Their expression was measured by qPCR at 4 time points in 27 de novo ALL patients treated in the ALL IC-BFM 2009 study. RESULTS: The expression of all 4 miRs significantly decreased over the first week of therapy (miR-128-3p: log2 fold change - 2.86; adjusted p 3.6 × 10-7; miR-181b-5p: log2 fold change - 1.75; adjusted p 1.48 × 10-2; miR-181a-5p: log2 fold change -1.33; adjusted p 3.12 × 10-2; miR-222-3p: log2 fold change - 1.25; adjusted p 1.66 × 10-2). However, no significant further reduction in miR expression was found after the 8th day of therapy. Measured drop in expression of 2 miRs at day 8 strongly correlated with day 15 bone marrow flow cytometry MRD results (miR-128-3p: Pearson's r = 0.88, adjusted p = 2.71 × 10-4; miR-222-3p: r = 0.81, adjusted p = 2.99 × 10-3). CONCLUSION: In conclusion, these circulating miRs might act as biomarkers of residual leukemia. MiR-128-3p and miR-222-3p in blood predict day 15 flow cytometry MRD results 7 days earlier. Although, their sensitivity falls behind that of bone marrow flow cytometry MRD at day 15.

Digital behavior surveillance: Monitoring dental caries and toothache interests of Google users from developing countries.

OBJECTIVE: To determine the differences between the levels of interest of South American Google users on information related to dental caries and toothache over time. MATERIALS AND METHODS: The monthly variation of Relative Search Volume (RSV) for both queries was obtained in Google Trends between 2004 and 2017. The association between RSVs, Internet penetration, and disability-adjusted life-years (DALYs) for dental caries in permanent teeth were tested by distinct regression models. Forecasting ARIMA models were developed to analyze trends and predict the interests until December 2018. RESULTS: In general, toothache-related searches presented an uptrend associated positively with Internet penetration and negatively with DALYs, whereas dental caries-related searches presented a downtrend associated negatively with Internet penetration and positively with DALYs. The seasonality affected only dental caries curves, with the highest rates observed in the first trimester and the lowest rates in June, July, and December. CONCLUSION: The levels of interests of South American Google users in seeking dental caries and toothache information appear to have a direct connection with the burden of untreated dental caries lesions and Internet penetration. The no association between dental caries- and toothache-related interests reinforces the unfamiliarity of most people about the relationship of dental pain with the final stages of chronic oral diseases.

Possible roles of genetic variations in chemotherapy related cardiotoxicity in pediatric acute lymphoblastic leukemia and osteosarcoma.

BACKGROUND: The treatment of acute lymphoblastic leukemia (ALL) and osteosarcoma (OSC) is very effective: the vast majority of patients recover and survive for decades. However, they still need to face serious adverse effects of chemotherapy. One of these is cardiotoxicity which may lead to progressive heart failure in the long term. Cardiotoxicity is contributed mainly to the use of anthracyclines and might have genetic risk factors. Our goal was to test the association between left ventricular function and genetic variations of candidate genes. METHODS: Echocardiography data from medical records of 622 pediatric ALL and 39 OSC patients were collected from the period 1989-2015. Fractional shortening (FS) and ejection fraction (EF) were determined, 70 single nucleotide polymorphisms (SNPs) in 26 genes were genotyped. Multivariate logistic regression and multi-adjusted general linear model were performed to investigate the influence of genetic polymorphisms on the left ventricular parameters. Bayesian network based Bayesian multilevel analysis of relevance (BN-BMLA) method was applied to test for the potential interaction of the studied cofactors and SNPs. RESULTS: Our results indicate that variations in ABCC2, CYP3A5, NQO1, SLC22A6 and SLC28A3 genes might influence the left ventricular parameters. CYP3A5 rs4646450 TT was 17% among ALL cases with FS lower than 28, and 3% in ALL patients without pathological FS (p = 5.60E-03; OR = 6.94 (1.76-27.39)). SLC28A3 rs7853758 AA was 12% in ALL cases population, while only 1% among controls (p = 6.50E-03; OR = 11.56 (1.98-67.45)). Patients with ABCC2 rs3740066 GG genotype had lower FS during the acute phase of therapy and 5-10 years after treatment (p = 7.38E-03, p = 7.11E-04, respectively). NQO1 rs1043470 rare T allele was associated with lower left ventricular function in the acute phase and 5-10 years after the diagnosis (p = 4.28E-03 and 5.82E-03, respectively), and SLC22A6 gene rs6591722 AA genotype was associated with lower mean FS (p = 1.71E-03), 5-10 years after the diagnosis. CONCLUSIONS: Genetic variants in transporters and metabolic enzymes might modulate the individual risk to cardiac toxicity after chemotherapy.

HLA-DRB1*07:01-HLA-DQA1*02:01-HLA-DQB1*02:02 haplotype is associated with a high risk of asparaginase hypersensitivity in acute lymphoblastic leukemia.

Hypersensitivity reactions are the most frequent dose-limiting adverse reactions to Escherichia coli-derived asparaginase in pediatric acute lymphoblastic leukemia (ALL) patients. The aim of the present study was to identify associations between sequence-based Human Leukocyte Antigen Class II region alleles and asparaginase hypersensitivity in a Hungarian ALL population. Four-digit typing of HLA-DRB1 and HLA-DQB1 loci was performed in 359 pediatric ALL patients by using next-generation sequencing method. Based on genotypic data of the two loci, haplotype reconstruction was carried out. In order to investigate the possible role of the HLA-DQ complex, the HLA-DQA1 alleles were also inferred. Multivariate logistic regression analysis and a Bayesian network-based approach were applied to identify relevant genetic risk factors of asparaginase hypersensitivity. Patients with HLA-DRB1*07:01 and HLA-DQB1*02:02 alleles had significantly higher risk of developing asparaginase hypersensitivity compared to non-carriers [P=4.56×10-5; OR=2.86 (1.73-4.75) and P=1.85×10-4; OR=2.99 (1.68-5.31); n=359, respectively]. After haplotype reconstruction, the HLA-DRB1*07:01-HLA-DQB1*02:02 haplotype was associated with an increased risk. After inferring the HLA-DQA1 alleles the HLA-DRB1*07:01-HLA-DQA1*02:01-HLA-DQB1*02:02 haplotype was associated with the highest risk of asparaginase hypersensitivity [P=1.22×10-5; OR=5.00 (2.43-10.29); n=257]. Significantly fewer T-cell ALL patients carried the HLA-DQB1*02:02 allele and the associated haplotype than did pre-B-cell ALL patients (6.5%; vs. 19.2%, respectively; P=0.047). In conclusion, we identified a haplotype in the Human Leukocyte Antigen Class II region associated with a higher risk of asparaginase hypersensitivity. Our results confirm that variations in HLA-D region might influence the development of asparaginase hypersensitivity.

Efficient synthesis of cysteine-rich cyclic peptides through intramolecular native chemical ligation of N-Hnb-Cys peptide crypto-thioesters.

We herein introduce a straightforward synthetic route to cysteine-containing cyclic peptides based on the intramolecular native chemical ligation of in situ generated thioesters. Key precursors are N-Hnb-Cys crypto-thioesters, easily synthesized by Fmoc-based SPPS. The strategy is applied to a representative range of naturally occurring cyclic disulfide-rich peptide sequences.