Development and launch of the first obstetrics and gynaecology master of medicine residency training programme in Botswana.

BACKGROUND: Sub-Saharan Africa (SSA) faces a severe shortage of Obstetrician Gynaecologists (OBGYNs). While the Lancet Commission for Global Surgery recommends 20 OBGYNs per 100,000 population, Botswana has only 40 OBGYNs for a population of 2.3 million. We describe the development of the first OBGYN Master of Medicine (MMed) training programme in Botswana to address this human resource shortage. METHODS: We developed a 4-year OBGYN MMed programme at the University of Botswana (UB) using the Kern's approach. In-line with UB MMed standards, the programme includes clinical apprenticeship training complemented by didactic and research requirements. We benchmarked curriculum content, learning outcomes, competencies, assessment strategies and research requirements with regional and international programmes. We engaged relevant local stakeholders and developed international collaborations to support in-country subspecialty training. RESULTS: The OBGYN MMed curriculum was completed and approved by all relevant UB bodies within ten months during which time additional staff were recruited and programme financing was assured. The programme was advertised immediately; 26 candidates applied for four positions, and all selected candidates accepted. The programme was launched in January 2020 with government salary support of all residents. The clinical rotations and curricular development have been rolled out successfully. The first round of continuous assessment of residents was performed and internal programme evaluation was conducted. The national accreditation process was initiated. CONCLUSION: Training OBGYNs in-country has many benefits to health systems in SSA. Curricula can be adjusted to local resource context yet achieve international standards through thoughtful design and purposeful collaborations.

Is Greek affiliation a prescription for drug abuse? Examining misuse of prescription stimulants and downers in high school and college.

Objective: Compared to nonaffiliated students, Greek students engage in higher rates of prescription drug abuse (PDA). We examined PDA among Greek students compared to non-Greek students, and whether PDA in high school predicted joining a Greek organization in college.Participants: From November to December 2014, 509 undergraduate students at a large, southeastern institution participated in an online survey.Methods: Stratified random sampling with academic classification as the strata; chi-square analyses were conducted to test study hypotheses.Results: Greek students were no more likely than non-Greek students to ever engage in overall PDA. However, Greek students were significantly more likely to report abuse of stimulants, but not downers, in college and in high school.Conclusions: Future research to examine individual types of prescription drugs abused in high school and their affiliation with PDA among Greek students, compared to non-Greek students, in college is warranted.

Integrating Optical Tweezers, DNA Tightropes, and Single-Molecule Fluorescence Imaging: Pitfalls and Traps.

Fluorescence imaging is one of the cornerstone techniques for understanding how single molecules search for their targets on DNA. By tagging individual proteins, it is possible to track their position with high accuracy. However, to understand how proteins search for targets, it is necessary to elongate the DNA to avoid protein localization ambiguities. Such structures known as "DNA tightropes" are tremendously powerful for imaging target location; however, they lack information about how force and load affect protein behavior. The use of optically trapped microstructures offers the means to apply and measure force effects. Here we describe a system that we recently developed to enable individual proteins to be directly manipulated on DNA tightropes. Proteins bound to DNA can be conjugated with Qdot fluorophores for visualization and also directly manipulated by an optically trapped, manufactured microstructure. Together this offers a new approach to understanding the physical environment of molecules, and the combination with DNA tightropes presents opportunities to study complex biological phenomena.

Whole-genome association study of antibody response to Epstein-Barr virus in an African population: a pilot.

Epstein Barr virus (EBV) infects 95% of the global population and is associated with up to 2% of cancers globally. Immunoglobulin G (IgG) antibody levels to EBV have been shown to be heritable and associated with developing malignancies. We, therefore, performed a pilot genome-wide association analysis of anti-EBV IgG traits in an African population, using a combined approach including array genotyping, whole-genome sequencing and imputation to a panel with African sequence data. In 1562 Ugandans, we identify a variant in human leukocyte antigen (HLA)-DQA1, rs9272371 (p = 2.6 × 10-17) associated with anti-EBV nuclear antigen-1 responses. Trans-ancestry meta-analysis and fine-mapping with European-ancestry individuals suggest the presence of distinct HLA class II variants driving associations in Uganda. In addition, we identify four putative, novel, very rare African-specific loci with preliminary evidence for association with anti-viral capsid antigen IgG responses which will require replication for validation. These findings reinforce the need for the expansion of such studies in African populations with relevant datasets to capture genetic diversity.

Evaluating environmental and social influences on iron and zinc status of pregnant subsistence farmers in two geographically contrasting regions of Southern Malawi.

Micronutrient deficiency affects over 4.5 billion people worldwide, the majority in developing countries. Deficiencies of iron (and associated anaemia) and zinc in pregnancy are associated with complications, maternal and neonatal mortality, and developmental disorders in the foetus and growing child. We report the results of pilot study which used an interdisciplinary approach to explore environmental and sociocultural factors influencing the micronutrient status in the soil-plant-human transfer for pregnant subsistence farmers in two geographically contrasting regions of Southern Malawi. It evaluated micronutrient status in soil and the staple crop and explored the context for their transfer to pregnant women. Scientific and social science methods were used to collect data, following full sensitisation of the communities. A total of 99 participants were recruited from Chiradzulu (plateau) and Chikwawa (floodplain). Soil, maize and blood samples were collected, along with food frequency and health behaviour questionnaires and anthropological observation. Statistical analysis revealed that soil iron was significantly higher in Chiradzulu than in Chikwawa; total iron concentration is not deemed to be deficient in either area. Soil zinc was not significantly different between areas. Maize concentrations of iron and zinc were not significantly different between areas, and were not deficient relative to improved cultivars. Blood iron deficiency and associated anaemia were problematic in both areas, but more so in Chikwawa than in Chiradzulu, and zinc deficiency was similar in both areas. The study has identified a significant difference in the blood iron status of the participants of the two communities, and has shown that this difference is not accounted for by the staple crop maize. Socio-geographical factors appear to play a significant role in the micronutrient health of the populations. The findings lend support to multifaceted community intervention studies which educate communities on strategies to tackle micronutrient deficiencies.

Anti-streptococcal, tubulin, and dopamine receptor 2 antibodies in children with PANDAS and Tourette syndrome: single-point and longitudinal assessments.

Single-point-in-time ELISA optical densities for three putative antibodies identified in Sydenham's chorea, the streptococcal group A carbohydrate antigen, N-acetyl-beta-d-glucosamine, tubulin, and the dopamine 2 receptor, showed no differences in children with PANDAS (n=44) or Tourette syndrome (n=40) as compared to controls (n=24). Anti-tubulin and D2 receptor antibodies assessed in serial samples from 12 PANDAS subjects obtained prior to a documented exacerbation, during the exacerbation (with or without a temporally associated streptococcal infection), and following the exacerbation, showed no evidence of antibody levels correlating with a clinical exacerbation. These data do not support hypotheses suggesting an autoimmune hypothesis in either TS or PANDAS.

Time-resolved multiple probe spectroscopy.

Time-resolved multiple probe spectroscopy combines optical, electronic, and data acquisition capabilities to enable measurement of picosecond to millisecond time-resolved spectra within a single experiment, using a single activation pulse. This technology enables a wide range of dynamic processes to be studied on a single laser and sample system. The technique includes a 1 kHz pump, 10 kHz probe flash photolysis-like mode of acquisition (pump-probe-probe-probe, etc.), increasing the amount of information from each experiment. We demonstrate the capability of the instrument by measuring the photolysis of tungsten hexacarbonyl (W(CO)(6)) monitored by IR absorption spectroscopy, following picosecond vibrational cooling of product formation through to slower bimolecular diffusion reactions on the microsecond time scale.

A tripartite complex composed of ETV6-NTRK3, IRS1 and IGF1R is required for ETV6-NTRK3-mediated membrane localization and transformation.

ETV6-NTRK3 (EN), a chimeric tyrosine kinase generated by t(12;15) translocations, is a dominantly acting oncoprotein in diverse tumor types. We previously showed that insulin-like growth factor 1 receptor (IGF1R) is essential for EN-mediated oncogenesis and that insulin receptor substrate 1 (IRS1) is constitutively tyrosine phosphorylated and bound by EN in transformed cells. Given that IRS1 is also an adapter for IGF1R, we hypothesized that IRS1 might localize EN to IGF1R at the membrane to activate phosphatidylinositol 3-kinase (PI3K)-Akt, which is critical for EN oncogenesis. In this study, we examined EN/IRS1/IGF1R complexes in detail. We find that both IRS1 and kinase active IGF1R are required for EN transformation, that tyrosine phosphorylated IRS1 is present in high molecular weight complexes with EN and IGF1R, and that EN colocalizes with IGF1R at the plasma membrane. Both IGF1R kinase activity and an intact cytoplasmic Y950 residue, the IRS1-docking site of IGF1R, are required, confirming the importance of the IGF1R/IRS1 interaction for EN oncogenesis. The dual specificity IGF1R and insulin receptor (INSR) inhibitor, BMS-536924, blocks EN transformation activity, cell survival and its interaction with IRS proteins, and induces a striking shift of EN proteins to smaller sized molecular complexes. We conclude that a tripartite complex of EN, IRS1 and IGF1R localizes EN to the membrane and that this is essential for EN-mediated transformation. These findings provide an explanation for the observed IGF1R dependency of EN transformation. Blocking IGF1R kinase activity may, therefore, provide a tractable therapeutic strategy for the many tumor types driven by the EN oncoprotein.

Rapid ultrathin slice plastination of embalmed specimens with minimal tissue loss.

A modified technique of producing 1 mm (ultrathin) E12 plastinated slices of tissue specimens from embalmed cadavers for anatomical teaching and research is described. Specimens up to 150 mm in length and width were embedded in polyurethane foam and serially sectioned using an OMAS C300 food slicer. Individual slices were then processed by cold dehydration, degreasing, resin impregnation, mounting and curing. Bone-containing specimens were first decalcified. Tissue slices could be cut with remarkable accuracy (0.98 +/- 0.01 mm per slice) and minimal tissue loss (less than 2% per slice). The entire production process could be achieved in 25 days and was associated with tissue volume shrinkage of no more than 6%. These results demonstrate a relatively rapid method of producing ultrathin E12 slices with minimal tissue loss. The technique may be particularly applicable to submacroscopic morphologic studies and three-dimensional reconstruction in clinical anatomy.

Dynamic position and force measurement for multiple optically trapped particles using a high-speed active pixel sensor.

A high frame rate active pixel sensor designed to track the position of up to six optically trapped objects simultaneously within the field of view of a microscope is described. The sensor comprises 520 x 520 pixels from which a flexible arrangement of six independent regions of interest is accessed at a rate of up to 20 kHz, providing the capability to measure motion in multiple micron scale objects to nanometer accuracy. The combined control of both the sensor and optical traps is performed using unique, dedicated electronics (a field programmable gate array). The ability of the sensor to measure the dynamic position and the forces between six optically trapped spheres, down to femtonewton level, is demonstrated paving the way for application in the physical and life sciences.

An audit of completeness of HIV clinical histories: before and after introduction of an HIV proforma.

The completeness of a 'first consultation' human immunodeficiency virus (HIV) clinical history before and after the introduction of an HIV proforma was audited by a retrospective case notes review. Twenty key variables considered essential to every history were assessed. There was a significant improvement in the documentation of 14 of the 18 items for men and 14 of the 20 items for women post-proforma with no deterioration in documentation of any of the variables. Our study supports the introduction of a structured 'first consultation' notes proforma for use during consultations with both newly diagnosed HIV-positive patients and those transferring their care from other centres.

Anxiety related to dental general anaesthesia: changes in anxiety in children and their parents.

AIM: To examine the anxiety levels of children referred for dental general anaesthesia and their parents at various key points of the referral and anaesthetic procedure. METHODS: Structured interviews and anxiety measures were conducted with 50 children attending the Department of Paediatric Dentistry, Leeds Dental Institute, and progressing to general anaesthetic (GA) and their parents. Interviews were conducted with parents and children prior to initial assessment, following assessment and prior to a GA. Anxiety was measured at each interview, using the Visual Analogue Scale for parents and the Venham's Picture test for children. A fourth telephone interview was conducted with parents one week after the GA when the degree of upset caused to parents and children by the procedure was evaluated. RESULTS: Anxiety of children remained constant at each interview. Parent and child anxiety were not related. There was a rise in parent anxiety following initial assessment in those families attending in response to a routine referral and progressing to GA (p<0.05). There was a further rise in parent anxiety in these families immediately prior to the GA itself (p<0.001). Parent upset was strongly related to their anxiety at each of the three interviews prior to the GA (p<0.01, 0.05 and 0.001 respectively) and to the distress of their child (p<0.02). Child distress was strongly related to anxiety at each of the three interviews prior to the GA. CONCLUSION: The anxiety levels of children did not appear to change throughout the whole GA assessment and treatment process. Parent anxiety rose significantly following assessment and again just prior to the GA. Factors contributing to parent upset post treatment were child upset and pre treatment parent anxiety levels. Children who were most anxious prior to GA found the procedure most distressing.

Effects of gravity, hypergravity and microgravity on vestibular neurones of the crab.

Recordings were made from identified balancing system interneurones using implanted electrodes in crabs oscillated at 0.3 Hz during bouts of Parabolic flight. Repeatable non stabilized patterns of response firing were seen in head up and head down interneurones. During the hypergravity phases, the ratio of firing frequencies in the two directional categories of interneurones was altered showing that hypergravity produced effects normally seen during tilting of the crab, implying greater bending of the sensory thread hairs. During microgravity, firing levels remained low and constant or changed slowly towards initial firing levels.

A prospective clinical trial comparing preformed metal crowns and cast restorations for defective first permanent molars.

AIM: This study investigated two methods for the restoration of permanent molars affected by amelogenesis imperfecta (AI) or severe enamel defects. METHODS: A prospective clinical trial was carried out on 17 subjects affected by AI or severe enamel defects of first permanent molars. A split mouth design was used so that each right or left permanent molar in both jaws was restored using either a preformed metal crown (SSC) or a cast adhesive coping (CAC). Subjects were followed for up to 24 months and assessed for longevity and quality of the restorations. Sequential analysis was used to compare longevity. RESULTS: Records for 42 restorations (19 SSC; 23 CAC) were kept. The split mouth design was possible on 24 occasions (right versus left=14; maxilla versus mandible=10). Three restorations, one SSC (at 6 months) and two CAC (at 2 and 19 months) failed and required replacement. There was no significant statistical difference between the two types of restorations. CONCLUSION: While there was no difference between the two restorations for quality and longevity, the SSC was considerably cheaper to use and needed only one visit, but more tooth tissue was lost in preparation and fitting. The CAC was significantly more expensive but left nearly all of the tooth crown intact. The choice of which restoration to use is indicated by the immediate and long-term needs of each individual patient.

Magnetic bead purification of labeled DNA fragments for high-throughput capillary electrophoresis sequencing.

We have developed an automated purification method for dye-terminator-based DNA sequencing products using a magnetic bead approach. This 384-well protocol generates sequence fragments that are essentially free of template DNA, salt, and excess dye-terminator products. In comparison with traditional ethanol precipitation protocols, this method uses no centrifugation, is rapid, completely automated, and increases the phred-20 read length by an average of 40 bases. To date, we have processed over 4 million samples with 94% averaging 641 phred-20 bases on the MegaBACE 1000 and 4000 and the ABI PRISM 3700 capillary instruments.

Diet and the duration of testosterone-dependent prostate cancer in Lobund-Wistar rats.

Hormone-related cancers in the prostate, breast, endometrium, ovary and testicle account for 30% of malignancies in humans. We have developed a unique model of spontaneous prostate cancer (PC) in Lobund-Wistar [L-W] rats that shares many of its characteristics with the natural history of PC in man, including (a) inherent predisposition, high production of testosterone and aging risk factors, (b) endogenous tumorigenic mechanisms, and (c) early stage testosterone-dependent and late stage testosterone-independent tumors. About 30% of L-W rats on diet L-485 develop spontaneous palpable cancer in the anterior prostate-seminal vesicle (P-SV) complex in average of 20.5 months. At age 12 months early stage spontaneous PC was prevented or reversed by testosterone-deprivation through change of diet from L-485 to soy protein isolate/isoflavone (SPII) diet, thereby preventing the late lethal clinical disease: about 75% of rats at risk of developing testosterone-independent P-SV tumors were free of detectable cancer and about 25% had developed testosterone-independent cancer at age 12 months. The duration of the dependent stage exceeded age 12 months in 75% of the rats at risk. Dietary soymeal, found in most natural ingredient diets, may promote PC tumorigenesis, but only in L-W rats.

Human chromosome 19 and related regions in mouse: conservative and lineage-specific evolution.

To illuminate the function and evolutionary history of both genomes, we sequenced mouse DNA related to human chromosome 19. Comparative sequence alignments yielded confirmatory evidence for hypothetical genes and identified exons, regulatory elements, and candidate genes that were missed by other predictive methods. Chromosome-wide comparisons revealed a difference between single-copy HSA19 genes, which are overwhelmingly conserved in mouse, and genes residing in tandem familial clusters, which differ extensively in number, coding capacity, and organization between the two species. Finally, we sequenced breakpoints of all 15 evolutionary rearrangements, providing a view of the forces that drive chromosome evolution in mammals.

Differential sympathetic nerve and heart rate spectral effects of nonhypotensive lower body negative pressure.

Lower body negative pressure (LBNP; -5 and -15 mmHg) was applied to 14 men (mean age 44 yr) to test the hypothesis that reductions in preload without effect on stroke volume or blood pressure increase selectively muscle sympathetic nerve activity (MSNA), but not the ratio of low- to high-frequency harmonic component of spectral power (P(L)/P(H)), a coarse-graining power spectral estimate of sympathetic heart rate (HR) modulation. LBNP at -5 mmHg lowered central venous pressure and had no effect on stroke volume (Doppler) or systolic blood pressure but reduced vagal HR modulation. This latter finding, a manifestation of arterial baroreceptor unloading, refutes the concept that low levels of LBNP interrogate, selectively, cardiopulmonary reflexes. MSNA increased, whereas P(L)/P(H) and HR were unchanged. This discordance is consistent with selectivity of efferent sympathetic responses to nonhypotensive LBNP and with unloading of tonically active sympathoexcitatory atrial reflexes in some subjects. Hypotensive LBNP (-15 mmHg) increased MSNA and P(L)/P(H), but there was no correlation between these changes within subjects. Therefore, HR variability has limited utility as an estimate of the magnitude of orthostatic changes in sympathetic discharge to muscle.