Single-cell profiling of Anopheles gambiae spermatogenesis defines the onset of meiotic silencing and premeiotic overexpression of the X chromosome.

Understanding development and genetic regulation in the Anopheles gambiae germline is essential to engineer effective genetic control strategies targeting this malaria mosquito vector. These include targeting the germline to induce sterility or using regulatory sequences to drive transgene expression for applications such as gene drive. However, only very few germline-specific regulatory elements have been characterised with the majority showing leaky expression. This has been shown to considerably reduce the efficiency of current genetic control strategies, which rely on regulatory elements with more tightly restricted spatial and/or temporal expression. Meiotic silencing of the sex chromosomes limits the flexibility of transgene expression to develop effective sex-linked genetic control strategies. Here, we build on our previous study, dissecting gametogenesis into four distinct cell populations, using single-cell RNA sequencing to define eight distinct cell clusters and associated germline cell-types using available marker genes. We reveal overexpression of X-linked genes in a distinct cluster of pre-meiotic cells and document the onset of meiotic silencing of the X chromosome in a subcluster of cells in the latter stages of spermatogenesis. This study provides a comprehensive dataset, characterising the expression of distinct cell types through spermatogenesis and widening the toolkit for genetic control of malaria mosquitoes.

A zebrafish model of growth hormone insensitivity syndrome with immune dysregulation 1 (GHISID1).

Signal transducer and activator of transcription (STAT) proteins act downstream of cytokine receptors to facilitate changes in gene expression that impact a range of developmental and homeostatic processes. Patients harbouring loss-of-function (LOF) STAT5B mutations exhibit postnatal growth failure due to lack of responsiveness to growth hormone as well as immune perturbation, a disorder called growth hormone insensitivity syndrome with immune dysregulation 1 (GHISID1). This study aimed to generate a zebrafish model of this disease by targeting the stat5.1 gene using CRISPR/Cas9 and characterising the effects on growth and immunity. The zebrafish Stat5.1 mutants were smaller, but exhibited increased adiposity, with concomitant dysregulation of growth and lipid metabolism genes. The mutants also displayed impaired lymphopoiesis with reduced T cells throughout the lifespan, along with broader disruption of the lymphoid compartment in adulthood, including evidence of T cell activation. Collectively, these findings confirm that zebrafish Stat5.1 mutants mimic the clinical impacts of human STAT5B LOF mutations, establishing them as a model of GHISID1.

Neural Entrainment to Musical Pulse in Naturalistic Music Is Preserved in Aging: Implications for Music-Based Interventions.

Neural entrainment to musical rhythm is thought to underlie the perception and production of music. In aging populations, the strength of neural entrainment to rhythm has been found to be attenuated, particularly during attentive listening to auditory streams. However, previous studies on neural entrainment to rhythm and aging have often employed artificial auditory rhythms or limited pieces of recorded, naturalistic music, failing to account for the diversity of rhythmic structures found in natural music. As part of larger project assessing a novel music-based intervention for healthy aging, we investigated neural entrainment to musical rhythms in the electroencephalogram (EEG) while participants listened to self-selected musical recordings across a sample of younger and older adults. We specifically measured neural entrainment to the level of musical pulse-quantified here as the phase-locking value (PLV)-after normalizing the PLVs to each musical recording's detected pulse frequency. As predicted, we observed strong neural phase-locking to musical pulse, and to the sub-harmonic and harmonic levels of musical meter. Overall, PLVs were not significantly different between older and younger adults. This preserved neural entrainment to musical pulse and rhythm could support the design of music-based interventions that aim to modulate endogenous brain activity via self-selected music for healthy cognitive aging.

Driving down malaria transmission with engineered gene drives.

The last century has witnessed the introduction, establishment and expansion of mosquito-borne diseases into diverse new geographic ranges. Malaria is transmitted by female Anopheles mosquitoes. Despite making great strides over the past few decades in reducing the burden of malaria, transmission is now on the rise again, in part owing to the emergence of mosquito resistance to insecticides, antimalarial drug resistance and, more recently, the challenges of the COVID-19 pandemic, which resulted in the reduced implementation efficiency of various control programs. The utility of genetically engineered gene drive mosquitoes as tools to decrease the burden of malaria by controlling the disease-transmitting mosquitoes is being evaluated. To date, there has been remarkable progress in the development of CRISPR/Cas9-based homing endonuclease designs in malaria mosquitoes due to successful proof-of-principle and multigenerational experiments. In this review, we examine the lessons learnt from the development of current CRISPR/Cas9-based homing endonuclease gene drives, providing a framework for the development of gene drive systems for the targeted control of wild malaria-transmitting mosquito populations that overcome challenges such as with evolving drive-resistance. We also discuss the additional substantial works required to progress the development of gene drive systems from scientific discovery to further study and subsequent field application in endemic settings.

Investigation of two suspected diarrhoeal-illness outbreaks in Northern Cape and KwaZulu-Natal provinces, South Africa, April-July 2013: The role of rotavirus.

BACKGROUND: Suspected diarrhoeal-illness outbreaks affecting mostly children < 5 years were investigated between May and July 2013 in Northern Cape province (NCP) and KwaZulu-Natal (KZN) province. This study describes the epidemiological, environmental and clinical characteristics and diarrhoeal-illnesses causative agent(s). METHODS: A descriptive cross-sectional study was conducted. Cases were patients presenting at healthcare facilities with diarrhoeal-illness between 09 April and 09 July 2013 in NCP and 01 May and 31 July 2013 in KZN. Laboratory investigations were performed on stools and water samples using microscopy, culture and sensitivity screening and molecular assays. RESULTS: A total of 953 cases including six deaths (case fatality rate [CFR]: 0.6%) were recorded in the Northern Cape province outbreak. Children < 5 years accounted for 58% of cases. Enteric viruses were detected in 51% of stools, with rotavirus detected in 43%. The predominant rotavirus strains were G3P[8] (45%) and G9P[8] (42%). Other enteric viruses were detected, with rotavirus co-infections (63%). No enteric pathogens detected in water specimens. KwaZulu-Natal outbreak: A total of 1749 cases including 26 deaths (CFR: 1.5%) were recorded. Children < 5 years accounted for 95% of cases. Rotavirus was detected in 55% of stools; other enteric viruses were detected, mostly as rotavirus co-infections. The predominant rotavirus strains were G2P[4] (54%) and G9P[8] (38%). CONCLUSION: Although source(s) of the outbreaks were not identified, the diarrhoeal-illnesses were community-acquired. It is difficult to attribute the outbreaks to one causative agent(s) because of rotavirus co-infections with other enteric pathogens. While rotavirus was predominant, the outbreaks coincided with the annual rotavirus season.

Recommendations for radiographers and radiation therapists drawn from an analysis of errors on Australian Radiation Incident Registers.

INTRODUCTION: The Australian Radiation Protection and Nuclear Safety Agency (ARPANSA) collect reported incidents for inclusion in the Australian Radiation Incident Register (ARIR), a database of radiation incident reports that occur within Australia. While the information on previous radiation incidents is available, there is little information on the lessons that can be learned from those past incidents to help prevent the same errors reoccurring. The aims of the study were to investigate what radiation incident registers are publicly available in Australia and to utilise the information contained within the ARIR and any other state or territory radiation protection authority registers to make recommendations for radiographers and radiation therapists to prevent future adverse events. METHODS: A search was conducted to locate what radiation incident registers within Australia were available to the public. All adverse events from 2003 to 2014 were compiled into a spreadsheet for analysis. An error-type classification taxonomy was used to classify the adverse events. Conclusions were drawn from the determined causes to make recommendations to change work practices in an attempt to prevent similar adverse events reoccurring. RESULTS: Incident registers were located from New South Wales, South Australia, Tasmania, Victoria and Western Australia. Radiography represented 76% (243) of the adverse events. A vast majority of the incidents were a failure to comply with time-out protocols (77%, 248). CONCLUSION: There are several radiation adverse event registers publicly available to utilise in Australia. All departments need to adopt and strictly adhere to time-out protocols. This in conjunction with the other recommendations in this article has the potential to dramatically reduce radiation adverse events.

Metabolic effects of incremental doses of intraperitoneal amino acids on automated peritoneal dialysis.

BACKGROUND: The use of amino acid (AA) dialysate to ameliorate protein-energy malnutrition has been limited by adverse metabolic effects. OBJECTIVE: We undertook this study to examine the acute metabolic effects of escalating doses of AAs delivered with lactate/bicarbonate dialysate on automated peritoneal dialysis (APD). PATIENTS AND METHODS: 12 APD patients were treated with conventional lactate-buffered dialysate (week 1), followed by lactate/bicarbonate-buffered dialysate (week 2), then 2 - 2.5 L 1.1% AA solution were added (week 3), and then an additional 2 - 2.5 L 1.1% AA were added (week 4). The primary outcomes were change in serum bicarbonate and pH, change in protein catabolic rate (PCR), and change in normalized ultrafiltration (milliliters/gram of carbohydrate infused). RESULTS: Serum bicarbonate rose from week 1 to week 2 (28.9 +/- 3.2 vs 26.9 +/- 4.1 mmol/L, p = 0.03). Addition of one bag of AAs led to a decline in plasma bicarbonate (26.9 +/- 2.1 vs 28.9 +/- 3.2 mmol/L, p < 0.01), which was further magnified by the addition of the second bag of AAs (23.8 +/- 2.7 vs 26.9 +/- 2.1 mmol/L, p < 0.01). Serum bicarbonate fell significantly by week 4 compared to week 1 (23.8 +/- 2.7 vs 26.9 +/- 3.2 mmol/L, p < 0.01) although there was no significant change in venous pH or PCR when week 4 was compared to week 1. Normalized ultrafiltration was stable for the first 3 weeks but rose significantly in week 4 compared to week 1 (5.32 +/- 2.30 vs 4.14 +/- 1.58 mL/g, p = 0.03). CONCLUSIONS: Higher doses of AAs mixed with newer bicarbonate/lactate dialysate on APD result in a small decrease in serum bicarbonate but improved normalized ultrafiltration. This merits further study as both a nutritional supplement and a glucose-sparing strategy.