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Abstract Background This study investigated the prevalence and predictors of episiotomy among puerperal women accessing postnatal care in the Kumasi Metropolis. Methods A cross-sectional survey was conducted among 1750 postnatal mothers who had spontaneous vaginal using a questionnaire. Descriptive statistics were conducted to show the rate of episiotomy. Chi-square and binary logistic regression analyses were conducted using Jamovi software version 2.5.3. Results The prevalence of episiotomy among the study population was 49.7% compared to the 10% of all deliveries recommended by World Health Organisation (WHO). Among those who had an episiotomy, 88.5% gave consent, and 77.6% required repairs. In the bivariate analysis educational status, age, perineal tearing, marital status, household monthly income, spontaneous vaginal deliveries and previous episiotomy had a significant association with episiotomy risk. In the multivariate analysis, married women, lower education levels, moderate household incomes, grand multiparous women, spontaneous vaginal delivery (SVD), instrumental deliveries, longer second stages of labour, and attending more antenatal care (ANC) sessions were all associated with a reduced likelihood of episiotomy. Higher one-minute Apgar scores and anemia also lowered the odds. Conversely, hypertensives, women with four or more previous vaginal deliveries, and those with a history of episiotomy were more likely to undergo the procedure. Conclusion Episiotomies conducted was high, indicating potential overuse compared to WHO’s recommendations. Antenatal attendance and timing should be prioritised to improve maternal preparation and outcomes. Policy interventions to reduce unnecessary episiotomies are needed. Midwifery education must also emphasise the importance of respectful maternity care.

Abstract Background Insecticide-treated nets (ITNs) are pivotal tools for malaria prevention in endemic regions like Ghana. Understanding the protective factors and barriers influencing ITN ownership and usage is crucial for designing effective interventions. A scoping review was conducted to identify studies exploring protective factors and barriers related to ITN ownership and usage. Methods This review followed the guidelines by Askey and O’Malley. Search was done in four major databases including PubMed, Science Direct, PubMed CENTRAL, and JSTOR. Additional searches were done in Google Scholar and Google. Peer-reviewed and grey literature were included. Results A total of 24 papers met the eligibility criteria and were included in the review. Included studies found regional disparities in ITN ownership and usage. Furthermore, included studies reported ownership rates between 97.8 and 28% and usage rates between 94 and 20%. Protective factors facilitating ITN ownership were marital status, higher educational attainment, higher income levels, and being aged 25 years or older. In contrast, the factors for its use included community-level campaigns advocating for ITN use and awareness, individuals with secondary education or higher and those residing in urban areas. Missed opportunities in free distribution exercises and the unavailability of subsidized ITNs at health facilities were barriers to ownership. Conclusion Understanding and addressing protective factors and barriers influencing ITN ownership and usage are crucial for enhancing malaria prevention strategies and achieving sustainable progress in combating malaria in endemic areas. Collaborative and evidence-based interventions are essential for addressing these challenges effectively.

Abstract Background A challenge facing many Academic Health Centers (AHCs) attempting to revise health professions education to include the impact of racism as a social and structural determinant of health (SSDoH) is a lack of broad faculty expertise to reinforce and avoid undermining learning modules addressing this topic. To encourage an institutional culture that is in line with new anti-racism instruction, we developed a six-part educational series on the history of racism in America and its impact on contemporary health inequities for teaching structural competency to health professions academicians. Methods We developed a six-hour elective continuing education (CE) series for faculty and staff with the following objectives: (1) describe and discuss race as a social construct; (2) describe and discuss the decolonization of the health sciences and health care; (3) describe and discuss the history of systemic racism and structural violence from a socio-ecological perspective; and (4) describe and discuss reconciliation and repair in biomedicine. The series was spread over a six-month period and each monthly lecture was followed one week later by an open discussion debriefing session. Attendees were assessed on their understanding of each objective before and after each series segment. Results We found significant increases in knowledge and understanding of each objective as the series progressed. Attendees reported that the series helped them grapple with their discomfort in a constructive manner. Self-selected attendees were overwhelmingly women (81.8%), indicating a greater willingness to engage with this material than men. Conclusions The series provides a model for AHCs looking to promote anti-racism and structural competency among their faculty and staff.

Abstract Background The COVID-19 pandemic demonstrated the vital need for research to inform policy decision-making and save lives. The Wales COVID-19 Evidence Centre (WCEC) was established in March 2021 and funded for two years, to make evidence about the impact of the pandemic and ongoing research priorities for Wales available and actionable to policy decision-makers, service leads and the public. Objectives We describe the approaches we developed and our experiences, challenges and future vision. Program implementation The centre operated with a core team, including a public partnership group, and six experienced research groups as collaborating partners. Our rapid evidence delivery process had five stages: 1. Stakeholder engagement (continued throughout all stages); 2. Research question prioritisation; 3. Bespoke rapid evidence review methodology in a phased approach; 4. Rapid primary research; and 5. Knowledge Mobilisation to ensure the evidence was available for decision-makers. Main achievements Between March 2021–23 we engaged with 44 stakeholder groups, completed 35 Rapid Evidence Reviews, six Rapid Evidence Maps and 10 Rapid Evidence Summaries. We completed four primary research studies, with three published in peer reviewed journals, and seven ongoing. Our evidence informed policy decision-making and was cited in 19 Welsh Government papers. These included pandemic infection control measures, the Action Plan to tackle gender inequalities, and Education Renew and Reform policy. We conducted 24 Welsh Government evidence briefings and three public facing symposia. Policy implications Strong engagement with stakeholder groups, a phased rapid evidence review approach, and primary research to address key gaps in current knowledge enabled high-quality efficient, evidence outputs to be delivered to help inform Welsh policy decision-making during the pandemic. We learn from these processes to continue to deliver evidence from March 2023 as the Health and Care Research Wales Evidence Centre, with a broader remit of health and social care, to help inform policy and practice decisions during the recovery phase and beyond.

Abstract Background Therapeutic communication (TC) promotes effective patient care, emotional wellbeing, and improves patient outcomes. The purpose of this review was to synthesise evidence on barriers and outcomes of TC between nurses and patients in Africa and to inform future studies and systematic reviews. Methods Search for the records was done in four major databases including PubMed, Science Direct, PubMed CENTRAL, and JSTOR. Additional searches were done in Google Scholar and Google. Results and findings from published studies and grey literature were included. A total of 16 articles met the eligibility criteria and were included in the review. This scoping review followed the guidelines by Askey and O’Malley. Results Barriers associated with TC were grouped under five main themes: sociodemographic factors, patient-related factors, nurse-related factors, environmental and health service-related. Age, and religious beliefs were the main sociodemographic factors that hindered TC while stress and inadequate knowledge and skills were identified among the nurse-related barriers to TC. Extreme weather conditions and mosquito infestation were environmental factors while lack of access to TC educational programmes on TC was a health service-related factor that interfered with TC. Both positive and negative outcomes of TC were also identified. Conclusion Continuous professional development on TC is necessary to improve nurses’ attitudes and update their skills to enable them to render culturally competent nursing care to patients.

Abstract Background When an individual is detained in hospital it is important that they maintain contact with their family, friends and communities as these can be helpful for their well-being and recovery. Maintaining these relationships is also important to unpaid carers (family or friends), but they can be strained by carers’ instigation of, or compliance with, the involuntary detention. Section 17 of the Mental Health Act (1983) in England and Wales allows for temporary leave from hospital, from an hour in the hospital grounds to going home for a few days. However, carers are not always involved in decisions around statutory s.17 leave, even where they are expected to support someone at home. This study aimed to explore how practice can be improved to better involve and support carers around s.17 leave. Methods Semi-structured interviews and focus groups were held with 14 unpaid carers and 19 mental health practitioners, including four Responsible Clinicians, in three sites in England in 2021. The research explored views on what works well for carers around s.17 leave, what could be improved and the barriers to such improvements. Transcripts were analysed using reflexive thematic analysis. Results Three themes were identified in the analysis: the need for carer support and the challenges surrounding provision; challenges with communication, planning and feedback around s.17 leave; and inconsistency in involving carers around s.17 leave. Permeating all themes was a lack of resources presenting as under-staffing, high demands on existing staff, and lack of time and capacity to work and communicate with carers. Conclusion Implications include the need for more funding for mental health services for both prevention and treatment; staff training to increase confidence with carers; and standardised guidance for practitioners on working with carers around s.17 leave to help ensure consistency in practice. The study concluded with the production of a ‘S.17 Standard’, a guidance document based on the research findings consisting of 10 steps for practitioners to follow to support the greater involvement and support of carers.

Abstract The generation of neuronal diversity remains incompletely understood. In Drosophila, the central brain is populated by neural stem cells derived from progenitors called neuroblasts (NBs). There are two types of NBs, type 1 and 2. T1NBs have a relatively simple lineage, whereas T2NBs expand and diversify the neural population with the generation of intermediate neural progenitors (INPs), contributing many neurons to the adult central complex, a brain region essential for navigation. However, it is not fully understood how neural diversity is created in T2NB and INP lineages. Imp, an RNA-binding protein, is expressed in T2NBs in a high-to-low temporal gradient, while the RNA-binding protein Syncrip forms an opposing gradient. It remains unknown if Imp expression is carried into INPs; whether it forms a gradient similar to NBs; and whether INP expression of Imp is required for generating neuronal identity or morphology. Here, we show that Imp/Syp are both present in INPs, but not always in opposing gradients. We find that newborn INPs adopt their Imp/Syp levels from their parental T2NBs; that Imp and Syp are expressed in stage-specific high-to-low gradients in INPs. In addition, there is a late INP pulse of Imp. We find that neurons born from old INPs (E-PG and PF-R neurons) have altered morphology following both Imp knock-down and Imp overexpression. We conclude that Imp functions in INPs and newborn neurons to determine proper neuronal morphology and central complex neuropil organization.

Abstract Background Globally, over 40 million lives have been claimed by HIV/AIDS. In Ghana, more than 350,000 people are living with HIV. Non-disclosure of HIV status is a major barrier to HIV/AIDS eradication; yet, little is known of the determinants of HIV status disclosure in resource limited settings in Africa like Ghana. Objective Determine the predictors of HIV status disclosure among people living with HIV (PLHIV) and stimulate policy discourse on support systems for self-disclosure in Africa. Methods This is a descriptive cross-sectional study among PLHIV (n = 181) in sub-Saharan Africa, specifically the Volta region of Ghana. Bivariate probit regression was run to determine factors associated with HIV status disclosure among PLHIV. Results HIV status self-disclosure was reported by 50% of the respondents; nearly 65% disclosed their status to non-family members and non-partners. Significant correlates of HIV status disclosure either to partners or non-partners were marital status, monthly income, type of occupation, and being divorced due to HIV status (p

Abstract The mechanisms that generate neural diversity during development remains largely unknown. Here, we use scRNA-seq methodology to discover new features of the Drosophila larval CNS across several key developmental timepoints. We identify multiple progenitor subtypes – both stem cell-like neuroblasts and intermediate progenitors – that change gene expression across larval development, and report on new candidate markers for each class of progenitors. We identify a pool of quiescent neuroblasts in newly hatched larvae and show that they are transcriptionally primed to respond to the insulin signaling pathway to exit from quiescence, including relevant pathway components in the adjacent glial signaling cell type. We identify candidate “temporal transcription factors” (TTFs) that are expressed at different times in progenitor lineages. Our work identifies many cell type specific genes that are candidates for functional roles, and generates new insight into the differentiation trajectory of larval neurons.

Abstract Background Opioid-related overdose deaths have surged in the USA over the last two decades. Overdose fatalities are preventable with the timely administration of naloxone. Syringe service programs (SSP) have pioneered community-based naloxone distribution through overdose prevention and naloxone distribution (OEND) programs. There is a dearth of information with regards to best practices for community-based OEND. Methods We utilized a modified Delphi approach to develop a set of best practices for OEND delivery. Starting with an initial list of best practices, we engaged 27 experts, in the field of OEND programming who reviewed, made recommendations for changes, and assigned a priority to each best practice. Results Two rounds of input resulted in a final list of 20 best practices organized into four categories. The mean priority scores ranged from 1.17 to 2.17 (range 1 to 3). The top 5 ranked best practices were ensuring that SSP participants have low barrier, consistent, needs-based access to naloxone and that there is ample naloxone available within communities. While the remaining fifteen best practices were deemed important, they had more to do with organizational culture and implementation climate. Conclusions Increasing community-based OEND delivery is essential to reduce opioid overdose deaths; however, it will be insufficient to add programs without an eye toward quality of implementation and fidelity to the model upon which the evidence is based. This list of best practices summarizes the consensus among OEND experts and can serve as a tool for SSPs providing OEND programming to improve services.

Abstract Background Intracerebral hemorrhage (ICH) is a significant cause of death and disabilities. Recently, cell therapies using mesenchymal stem cells have been shown to improve ICH-induced neurobehavioral deficits. Based on these findings, we designed this study to evaluate the therapeutic efficacy and underlying mechanisms by which human amnion-derived stem cells (hAMSCs) would ameliorate neurobehavioral deficits of ICH-bearing hosts. Methods hAMSCs were induced from amnia obtained by cesarean section and administered intravenously to ICH-bearing mice during the acute phase. The mice were then subject to multitask neurobehavioral tests at the subacute phase. We attempted to optimize the dosage and timing of the hAMSC administrations. In parallel with the hAMSCs, a tenfold higher dose of human adipose-derived stem cells (ADSCs) were used as an experimental control. Specimens were obtained from the ICH lesions to conduct immunostaining, flow cytometry, and Western blotting to elucidate the underlying mechanisms of the hAMSC treatment. Results The intravenous administration of hAMSCs to the ICH-bearing mice effectively improved their neurobehavioral deficits, particularly when the treatment was initiated at Day 1 after the ICH induction. Of note, the hAMSCs promoted clinical efficacy equivalent to or better than that of hADSCs at 1/10 the cell number. The systemically administered hAMSCs were found in the ICH lesions along with the local accumulation of macrophages/microglia. In detail, the hAMSC treatment decreased the number of CD11b+CD45+ and Ly6G+ cells in the ICH lesions, while splenocytes were not affected. Moreover, the hAMSC treatment decreased the number of apoptotic cells in the ICH lesions. These results were associated with suppression of the protein expression levels of macrophage-related factors iNOS and TNFα. Conclusions Intravenous hAMSC administration during the acute phase would improve ICH-induced neurobehavioral disorders. The underlying mechanism was suggested to be the suppression of subacute inflammation and apoptosis by suppressing macrophage/microglia cell numbers and macrophage functions (such as TNFα and iNOS). From a clinical point of view, hAMSC-based treatment may be a novel strategy for the treatment of ICH.

Abstract Background Ghana has a generalized HIV epidemic and efforts have been made to curb the spread and reverse its effects on the general population. In the wake of COVID-19 pandemic, the health system was unsettled and antiretroviral therapy (ART) care has been impacted in diverse ways. The study sought to explore the effects of COVID-19 on ART service provision in Ghana from the perspectives of the healthcare workers. Methods An exploratory-descriptive qualitative approach was employed in this study. Using maximum variation sampling method, fifteen healthcare workers; nurses, data managers and pharmacists were recruited from an ART clinic in a Teaching Hospital in Ghana. In-depth interviews were conducted and analysed using Braun and Clarke thematic approach. Results Three themes emerged from the data; “… And the pandemic struck”, “Impact of the pandemic on ART service delivery”; “Effecting the needed change”. The healthcare workers’ initial reactions to the pandemic and their show of commitment in ensuring continued ART service was evident. COVID-19 impacted service delivery in three main ways; (1) clients’ clinic attendance was erratic at various stages of the pandemic, (2) irregular resource availability as shortage was reported due to affected last mile delivery as a result of the lockdown in Accra, and (3) the health worker-patient interaction became less engaging because of established COVID-19 protocols. The healthcare workers however instituted strategies such as adjusting the patient appointment schedule, health professionals’ work schedule, establishing several work stations, task-shifting, and ensuring the implementation of all the COVID-19 protocols within the ART unit to ensure consistent service delivery as well as patient and staff safety. The study also found a decline in the implementation of several strategies established in the ART clinic during the initial phases of the pandemic such as a decline in the supply of Personal Protective Equipment (PPEs) by hospital management. Conclusions Although several strategies were implemented to manage the effects of the pandemic on ART care, there is a need to establish pathways of support for healthcare workers within the ART clinic and to consolidate as well as institutionalise the changes that ensured continuous but safe service delivery.

Abstract Background The Performance and Fitness (PERF-FIT) test battery for children is a recently developed, valid assessment tool for measuring motor skill-related physical fitness in 5 to 12-year-old children living in low-income settings. The aim of this study was to determine: (1) inter-rater reliability and (2) test-retest reliability of the PERF-FIT in children from 3 different countries (Ghana, South Africa and the Netherlands). Method For inter-rater reliability 29 children, (16 boys and 13 girls, 6–10 years) were scored by 2 raters simultaneously. For test–retest reliability 72 children, (33 boys and 39 girls, 5–12 years) performed the test twice, minimally 1 week and maximally 2 weeks apart. Relative and absolute reliability indices were calculated. ANOVA was used to examine differences between the three assessor teams in the three countries. Results The PERF-FIT demonstrated excellent inter-rater reliability (ICC, 0.99) and good test-retest reliability (ICC, ≥ 0.80) for 11 of the 12 tasks, with a poor ICC for the Jumping item, due to low spread in values. A significant difference between first and second test occasion was present on half of the items, but the differences were small (Cohen’s d 0.01–0.17), except for Stepping, Side jump and Bouncing and Catching (Cohen’s d 0.34, 0.41 and 0.33, respectively). Overall, measurement error, Limits of Agreement and Coefficient of Variation had acceptable levels to support clinical use. No systematic dissimilarities in error were found between first and second measurement between the three countries but for one item (Overhead throw). Conclusions The PERF-FIT can reliably measure motor skill related fitness in 5 to 12-year-old children in different settings and help clinicians monitor levels of fundamental motor skills (throwing, bouncing, catching, jumping, hopping and balance), power and agility.

Abstract Neuronal networks are capable of undergoing rapid structural and functional changes called plasticity, which are essential for shaping circuit function during nervous system development. These changes range from short-term modifications on the order of milliseconds, to long-term rearrangement of neural architecture that could last for the lifetime of the organism. Neural plasticity is most prominent during development, yet also plays a critical role during memory formation, behavior, and disease. Therefore, it is essential to define and characterize the mechanisms underlying the onset, duration, and form of plasticity. Astrocytes, the most numerous glial cell type in the human nervous system, are integral elements of synapses and are components of a glial network that can coordinate neural activity at a circuit-wide level. Moreover, their arrival to the CNS during late embryogenesis correlates to the onset of sensory-evoked activity, making them an interesting target for circuit plasticity studies. Technological advancements in the last decade have uncovered astrocytes as prominent regulators of circuit assembly and function. Here, we provide a brief historical perspective on our understanding of astrocytes in the nervous system, and review the latest advances on the role of astroglia in regulating circuit plasticity and function during nervous system development and homeostasis.

Abstract Background Developing cortical neurons express a tightly choreographed sequence of cytoskeletal and transmembrane proteins to form and strengthen specific synaptic connections during circuit formation. Nectin-3 is a cell-adhesion molecule with previously described roles in synapse formation and maintenance. This protein and its binding partner, nectin-1, are selectively expressed in upper-layer neurons of mouse visual cortex, but their role in the development of cortical circuits is unknown. Methods Here we block nectin-3 expression (via shRNA) or overexpress nectin-3 in developing layer 2/3 visual cortical neurons using in utero electroporation. We then assay dendritic spine densities at three developmental time points: eye opening (postnatal day (P)14), one week following eye opening after a period of heightened synaptogenesis (P21), and at the close of the critical period for ocular dominance plasticity (P35). Results Knockdown of nectin-3 beginning at E15.5 or ~ P19 increased dendritic spine densities at P21 or P35, respectively. Conversely, overexpressing full length nectin-3 at E15.5 decreased dendritic spine densities when all ages were considered together. The effects of nectin-3 knockdown and overexpression on dendritic spine densities were most significant on proximal secondary apical dendrites. Interestingly, an even greater decrease in dendritic spine densities, particularly on basal dendrites at P21, was observed when we overexpressed nectin-3 lacking its afadin binding domain. Conclusion These data collectively suggest that the proper levels and functioning of nectin-3 facilitate normal synapse formation after eye opening on apical and basal dendrites in layer 2/3 of visual cortex.

Abstract Background Spatial patterning specifies neural progenitor identity, with further diversity generated by temporal patterning within individual progenitor lineages. In vertebrates, these mechanisms generate “cardinal classes” of neurons that share a transcription factor identity and common morphology. In Drosophila, two cardinal classes are Even-skipped (Eve)+ motor neurons projecting to dorsal longitudinal muscles, and Nkx6+ motor neurons projecting to ventral oblique muscles. Cross-repressive interactions prevent stable double-positive motor neurons. The Drosophila neuroblast 7–1 (NB7–1) lineage uses a temporal transcription factor cascade to generate five distinct Eve+ motor neurons; the origin and development of Nkx6+ motor neurons remains unclear. Methods We use a neuroblast specific Gal4 line, sparse labelling and molecular markers to identify an Nkx6+ VO motor neuron produced by the NB7–1 lineage. We use lineage analysis to birth-date the VO motor neuron to the Kr+ Pdm+ neuroblast temporal identity window. We use gain- and loss-of-function strategies to test the role of Kr+ Pdm+ temporal identity and the Nkx6 transcription factor in specifying VO neuron identity. Results Lineage analysis identifies an Nkx6+ neuron born from the Kr+ Pdm+ temporal identity window in the NB7–1 lineage, resulting in alternation of cardinal motor neuron subtypes within this lineage (Eve>Nkx6 > Eve). Co-overexpression of Kr/Pdm generates ectopic VO motor neurons within the NB7–1 lineage – the first evidence that this TTF combination specifies neuronal identity. Moreover, the Kr/Pdm combination promotes Nkx6 expression, which itself is necessary and sufficient for motor neuron targeting to ventral oblique muscles, thereby revealing a molecular specification pathway from temporal patterning to cardinal transcription factor expression to motor neuron target selection. Conclusions We show that one neuroblast lineage generates interleaved cardinal motor neurons fates; that the Kr/Pdm TTFs form a novel temporal identity window that promotes expression of Nkx6; and that the Kr/Pdm > Nkx6 pathway is necessary and sufficient to promote VO motor neuron targeting to the correct ventral muscle group.

Abstract Background The Beck Depression Inventory II (BDI-II) has been accepted as one of the most commonly used self-report measures for depression. However, there is no study examining the psychometric properties of the BDI-II among substance users in low- and middle-income countries such as Myanmar. Therefore, this study aimed to evaluate the suitability of using this instrument in substance users and to examine the reliability and diagnostic efficiency to be compared with the International Classification of Diseases (ICD-10) among substance users in Northern Shan State of Myanmar. Methods A respondent-driven sampling (RDS) method was applied for recruiting the participants in this study, and total 230 substance users were recruited. On the other hand, 50 participants who visited the drop-in center (DIC) were screened for depressive symptoms using ICD-10 during the days when one consultant psychiatrist doctor was presented. These participants were interviewed face-to-face by the consultant psychiatrist using a semi-structured questionnaire including the Myanmar version of the BDI-II (mBDI-II). Results The mBDI-II showed moderate accuracy with an area under the curve of 0.68. The optimal cutoff score was 10 with the highest Youden index (0.48), and it had high sensitivity and specificity (0.78 and 0.70). The Cronbach’s alpha coefficients for clinically depressed and non-clinically depressed substance users were 0.91 and 0.93, respectively. Confirmatory factor analysis of the mBDI-II indicated that a three-factor solution (cognitive, affective, and somatic) was the best fit for substance users. Conclusions The mBDI-II has sound psychometric properties among substance users with moderate accuracy and range of possible cutoff scores together with sensitivity and specificity.

Abstract More than 30 years of studies into Drosophila melanogaster neurogenesis have revealed fundamental insights into our understanding of axon guidance mechanisms, neural differentiation, and early cell fate decisions. What is less understood is how a group of neurons from disparate anterior-posterior axial positions, lineages and developmental periods of neurogenesis coalesce to form a functional circuit. Using neurogenetic techniques developed in Drosophila it is now possible to study the neural substrates of behavior at single cell resolution. New mapping tools described in this review, allow researchers to chart neural connectivity to better understand how an anatomically simple organism performs complex behaviors.

Abstract Background Fresh coconut (Cocos nucifera L) water is a clear, sterile, colourless, slightly acidic and naturally flavoured drink, mostly consumed in tropical areas. It is a rich source of nutrients and has been used for medical purposes. This study was designed to investigate changes in selected characteristics of coconut water after autoclaving, gamma irradiation and storage. Also, the study was designed for assessing the possibility of measuring the growth of bacterial in fresh, stored or sterilised coconut water using turbidity measurements (at wavelengths between 600 nm and 800 nm) or by dry biomass determinations. Results Portions of coconut water aseptically extracted from the matured fruit, (average pH of 6.33 ± 0.17) were either stored at 4°C, autoclaved at 121°C for 20 min., or irradiated with gamma rays at 5 kGy. Subsequent changes in selected characteristics were determined. Autoclaving, gamma irradiation and long term storage of coconut water at 4°C resulted both in the development of a pale to intense yellow colour and changes in turbidity. After storage, the dry matter content of fresh, autoclaved and irradiated coconut water by 52.0%, 23.5% and 5.0% respectively. There were also significant differences in the UV spectra before and after sterilisation and during the storage of the coconut water. Although changes in total carbohydrates were observed, they were not significant (p > 0.05). Conclusions The enormous differences in the characteristics before and after storage suggests that the use of turbidity and dry biomass measurements for measuring the growth of bacteria in fresh, autoclaved and gamma irradiated coconut water before storage is practicable without any possibility of interference by the innate turbidity, colour and dry matter of the coconut water. However, this is not practicable after storing the coconut waters at 4°C, since there were increases in the turbidity and dry matter of the coconut water to levels that will mask the turbidity of a growing bacteria culture.

Abstract Tissue homeostasis depends on the ability of stem cells to properly regulate self-renewal versus differentiation. Drosophila neural stem cells (neuroblasts) are a model system to study self-renewal and differentiation. Recent work has identified two types of larval neuroblasts that have different self-renewal/differentiation properties. Type I neuroblasts bud off a series of small basal daughter cells (ganglion mother cells) that each generate two neurons. Type II neuroblasts bud off small basal daughter cells called intermediate progenitors (INPs), with each INP generating 6 to 12 neurons. Type I neuroblasts and INPs have nuclear Asense and cytoplasmic Prospero, whereas type II neuroblasts lack both these transcription factors. Here we test whether Prospero distinguishes type I/II neuroblast identity or proliferation profile, using several newly characterized Gal4 lines. We misexpress prospero using the 19H09-Gal4 line (expressed in type II neuroblasts but no adjacent type I neuroblasts) or 9D11-Gal4 line (expressed in INPs but not type II neuroblasts). We find that differential prospero expression does not distinguish type I and type II neuroblast identities, but Prospero regulates proliferation in both type I and type II neuroblast lineages. In addition, we use 9D11 lineage tracing to show that type II lineages generate both small-field and large-field neurons within the adult central complex, a brain region required for locomotion, flight, and visual pattern memory.

Abstract Background It is well known that many anurans do not reproduce easily in captivity. Some methods are based on administration of mammalian hormones such as human chorionic gonadotropin, which are not effective in many frogs. There is a need for simple, cost-effective alternative techniques to induce spawning. Methods Our new method is based on the injection of a combination of a gonadotropin-releasing hormone (GnRH) agonist and a dopamine antagonist. We have named this formulation AMPHIPLEX, which is derived from the combination of the words amphibian and amplexus. This name refers to the specific reproductive behavior of frogs when the male mounts and clasps the female to induce ovulation and to fertilize the eggs as they are laid. Results We describe the use of the method and demonstrate its applicability for captive breeding in 3 different anuran families. We tested several combinations of GnRH agonists with dopamine antagonists using Lithobates pipiens. The combination of des-Gly10, D-Ala6, Pro-LHRH (0.4 microrams/g body weight) and metoclopramide (10 micrograms/g BWt. MET) was most effective. It was used in-season, after short-term captivity and in frogs artificially hibernated under laboratory conditions. The AMPHIPLEX method was also effective in 3 Argentinian frogs, Ceratophrys ornata, Ceratophrys cranwelli and Odontophrynus americanus. Conclusion Our approach offers some advantages over other hormonally-based techniques. Both sexes are injected only once and at the same time, reducing handling stress. AMPHIPLEX is a new reproductive management tool for captive breeding in Anura.

Abstract Background The emergence of drug resistant strains of Mycobacterium tuberculosis complex has made the management of tuberculosis difficult. Also, Mycobacterium species has a peculiar cell wall, made of an impermeable complex structure rich in mycolate, making the lyses of its cell difficult. In order to apply a radio-labelled-probe based detection of mutations in selected genes leading to drug resistance, we concede that the evaluation and modifications of nucleic acid extraction protocols that are less sophisticated and less prone to contamination would be useful in the management of tuberculosis in a resource-constrained setting. Findings The average amount of nucleic acids was determined for different extraction treatments. High temperature treatment only, yielded the lowest amount of nucleic acids, i.e. 15.7 ± 3.2 μg. The average amount of nucleic acids obtained with the addition of TE and triton-X100, was 133.7 ± 8.9 μg, while that obtained with the addition of TE only, and TE and SDS were 68.4 ± 22.7 μg and 70.4 ± 20.3 μg respectively. Other treatments yielded 28.8 ± 6.7 μg, 32.5 ± 2.4 μg and 36.9 ± 15.5 μg. The average amount of nucleic acids obtained with high temperature treatment in TE, and that obtained by freezing prior to high temperature treatment, successfully amplified for the genes of interest (rpoB, KatG, rrs). Conclusion We strongly recommend the use of 1× TE buffer, and freezing and heating for improved lysis of cultured M. tuberculosis, and therefore, as an effective method for the preparation of M. tuberculosis nucleic acid useful for PCR.

Abstract Background Proper neuronal function depends on forming three primary subcellular compartments: axons, dendrites, and soma. Each compartment has a specialized function (the axon to send information, dendrites to receive information, and the soma is where most cellular components are produced). In mammalian neurons, each primary compartment has distinctive molecular and morphological features, as well as smaller domains, such as the axon initial segment, that have more specialized functions. How neuronal subcellular compartments are established and maintained is not well understood. Genetic studies in Drosophila have provided insight into other areas of neurobiology, but it is not known whether flies are a good system in which to study neuronal polarity as a comprehensive analysis of Drosophila neuronal subcellular organization has not been performed. Results Here we use new and previously characterized markers to examine Drosophila neuronal compartments. We find that: axons and dendrites can accumulate different microtubule-binding proteins; protein synthesis machinery is concentrated in the cell body; pre- and post-synaptic sites localize to distinct regions of the neuron; and specializations similar to the initial segment are present. In addition, we track EB1-GFP dynamics and determine microtubules in axons and dendrites have opposite polarity. Conclusion We conclude that Drosophila will be a powerful system to study the establishment and maintenance of neuronal compartments.

Abstract Background The choice of a stem cell to divide symmetrically or asymmetrically has profound consequences for development and disease. Unregulated symmetric division promotes tumor formation, whereas inappropriate asymmetric division affects organ morphogenesis. Despite its importance, little is known about how spindle positioning is regulated. In some tissues cell fate appears to dictate the type of cell division, whereas in other tissues it is thought that stochastic variation in spindle position dictates subsequent sibling cell fate. Results Here we investigate the relationship between neural progenitor identity and spindle positioning in the Drosophila optic lobe. We use molecular markers and live imaging to show that there are two populations of progenitors in the optic lobe: symmetrically dividing neuroepithelial cells and asymmetrically dividing neuroblasts. We use genetically marked single cell clones to show that neuroepithelial cells give rise to neuroblasts. To determine if a change in spindle orientation can trigger a neuroepithelial to neuroblast transition, we force neuroepithelial cells to divide along their apical/basal axis by misexpressing Inscuteable. We find that this does not induce neuroblasts, nor does it promote premature neuronal differentiation. Conclusion We show that symmetrically dividing neuroepithelial cells give rise to asymmetrically dividing neuroblasts in the optic lobe, and that regulation of spindle orientation and division symmetry is a consequence of cell type specification, rather than a mechanism for generating cell type diversity.