Abnormal cleavage up to Day 3 does not compromise live birth and neonatal outcomes of embryos that have achieved full blastulation: a retrospective cohort study.

STUDY QUESTION: Do embryos displaying abnormal cleavage (ABNCL) up to Day 3 have compromised live birth rates and neonatal outcomes if full blastulation has been achieved prior to transfer? SUMMARY ANSWER: ABNCL is associated with reduced full blastulation rates but does not impact live birth rates and neonatal outcomes once full blastulation has been achieved. WHAT IS KNOWN ALREADY?: It is widely accepted that ABNCL is associated with reduced implantation rates of embryos when transferred at the cleavage stage. However, evidence is scarce in the literature reporting birth outcomes from blastocysts arising from ABNCL embryos, likely because they are ranked low priority for transfer. STUDY DESIGN, SIZE, DURATION: This retrospective cohort study included 1562 consecutive autologous in vitro fertilization cycles (maternal age 35.1 ± 4.7 years) performed at Fertility North, Australia between January 2017 and June 2022. Fresh transfers were performed on Day 3 or 5, with remaining embryos cultured up to Day 6 before vitrification. A total of 6019 embryos were subject to blastocyst culture, and a subset of 664 resulting frozen blastocysts was included for live birth and neonatal outcome analyses following single transfers. PARTICIPANTS/MATERIALS, SETTING, METHODS: ABNCL events were annotated from the first mitotic division up to Day 3, including direct cleavage (DC), reverse cleavage (RC) and <6 intercellular contact points at the 4-cell stage (<6ICCP). For DC and RC in combination, the ratios of affected blastomeres over the total number of all blastomeres up to Day 3 were also recorded. All pregnancies were followed up until birth with gestational age, birthweight, and sex of the baby being recorded. MAIN RESULTS AND THE ROLE OF CHANCE: Full blastulation rates for embryos showing DC (19.5%), RC (41.7%), <6ICCP (58.8%), and mixed (≥2) ABNCL types (26.4%) were lower than the rates for those without ABNCL (67.2%, P < 0.01 respectively). Subgroup analysis showed declining full blastulation rates with increasing ratios of combined DC/RC affected blastomeres over all blastomeres up to the 8-cell stage (66.2% when 0 affected, 47.0% when 0.25 affected, 27.4% when 0.5 affected, 14.5% when 0.75 affected, and 7.7% when all affected, P < 0.01). However, once full blastulation had been achieved, no difference was detected between DC, RC, <6ICCP, and no ABNCL blastocysts following single frozen transfers in subsequent live birth rates (25.9%, 33.0%, 36.0% versus 30.8%, P > 0.05, respectively), gestational age (38.7 ± 1.6, 38.5 ± 1.2, 38.3 ± 3.5 versus 38.5 ± 1.8 weeks, P > 0.05, respectively) and birthweight (3343.0 ± 649.1, 3378.2 ± 538.4, 3352.6 ± 841.3 versus 3313.9 ± 509.6 g, P > 0.05, respectively). Multiple regression (logistic or linear as appropriate) confirmed no differences in all of the above measures after accounting for potential confounders. LIMITATIONS, REASONS FOR CAUTION: Our study is limited by its retrospective nature, making it impossible to control every known or unknown confounder. Embryos in our dataset, being surplus after selection for fresh transfer, may not represent the general embryo population. WIDER IMPLICATIONS OF THE FINDINGS: Our findings highlight the incremental impact of ABNCL, depending on the ratio of affected blastomeres up to Day 3, on subsequent full blastulation. The reassuring live birth and neonatal outcomes of ABNCL blastocysts imply a potential self-correction mechanism among those embryos reaching the blastocyst stage, which provides valuable guidance for clinical practice and patient counseling. STUDY FUNDING/COMPETTING INTEREST(S): This research is supported by an Australian Government Research Training Program (RTP) Scholarship. All authors report no conflict of interest. TRIAL REGISTRATION NUMBER: N/A.

Inclusion of genital, sexual, and gender diversity in human reproductive teaching: impact on student experience and recommendations for tertiary educators.

Western societal norms have long been constrained by binary and exclusionary perspectives on matters such as infertility, contraception, sexual health, sexuality, and gender. These viewpoints have shaped research and knowledge frameworks for decades and led to an inaccurate and incomplete reproductive biology curriculum. To combat these deficiencies in reproductive systems-related education, our teaching team undertook a gradual transformation of unit content from 2018 to 2023, aiming to better reflect real diversity in human reproductive biology. This initiative involved intentional modifications, including clear use of pronoun self-identification by staff. We addressed the historical lack of representation of genital variation and helped students interrogate oversimplified reproductive biology binaries. A novel assignment was also introduced, prompting students to apply reproductive physiology knowledge to propose innovative assisted reproductive technology solutions for diverse demographics. The collective impact of these innovations had a positive effect on student learning. With improved lecture content and inclusive language, the proportion of inclusive group assignment topics chosen by students more than doubled in 2021. By 2022, coinciding with assessment topic changes, the percentage of inclusive assignments topics surpassed 50%. Further development of laboratory activities on intersex genital variation and genital modification raised further understanding of genital, sexual, gender, and cultural diversity. While implementing these changes posed challenges, pushing both staff and students out of their comfort zones at times, collaboration with relevant organizations and individuals with lived experience of queer identity proved integral. Ultimately, these relatively simple adjustments had a substantial impact on student experiences and appreciation for diversity.NEW & NOTEWORTHY We outline the teaching innovations that we have implemented to improve inclusion of diversity in reproductive biology and physiology contexts. This includes improved representation of genital, sexual, and gender diversity considerations in the curriculum. There is a critical need for these innovations as how we teach fundamentally shapes the understanding of our future medical and health professionals and researchers and thus influences the quality of future medical care and research.

Racial and ethnic disparities in major adverse limb events persist for chronic limb threatening ischemia despite presenting limb threat severity after peripheral vascular intervention.

OBJECTIVE: Racial and ethnic disparities have been well-documented in the outcomes for chronic limb threatening ischemia (CLTI). One purported explanation has been the disease severity at presentation. We hypothesized that the disparities in major adverse limb events (MALE) after peripheral vascular intervention (PVI) for CLTI would persist despite controlling for disease severity at presentation using the WIfI (Wound, Ischemia, foot Infection) stage. METHODS: The Vascular Quality Initiative PVI dataset (2016-2021) was queried for CLTI. Patients were excluded if they were missing the WIfI stage. The primary end point was the incidence of 1-year MALE, defined as major amputation (through the tibia or fibula or more proximally) or reintervention (endovascular or surgical) of the initial treatment limb. A multivariate hierarchical Fine-Gray analysis was performed, controlling for hospital variation, competing risk of death, and presenting WIfI stage, to assess the independent association of Black/African American race and Latinx/Hispanic ethnicity with MALE. A Cox proportional hazard regression model was used for the 1-year survival analysis. RESULTS: Overall, 47,830 patients (60%) had had WIfI scores reported (73% White, 20% Black, and 7% Latinx). The 1-year unadjusted cumulative incidence of MALE was 13.1% (95% confidence interval [CI], 12.6%-13.5%) for White, 14.3% (95% CI, 13.5%-15.3%) for Black, and 17.0% (95% CI, 15.3%-18.9%) for Latinx patients. On bivariate analysis, the occurrence of MALE was significantly associated with younger age, Black race, Latinx ethnicity, coronary artery disease, cerebrovascular disease, congestive heart failure, hypertension, diabetes, dialysis, intervention level, any prior minor or major amputation, and WIfI stage (P < .001). The cumulative incidence of 1-year MALE increased by increasing WIfI stage: stage 1, 11.7% (95% CI, 10.9%-12.4%); stage 2, 12.4% (95% CI, 11.8%-13.0%); stage 3, 14.8% (95% CI, 13.8%-15.8%); and stage 4, 15.4% (95% CI, 14.3%-16.6%). The cumulative incidence also increased by intervention level: inflow, 10.7% (95% CI, 9.8%-11.7%), femoropopliteal, 12.3% (95% CI, 11.7%-12.9%); and infrapopliteal, 14.1% (95% CI, 13.5%-14.8%). After adjustment for WIfI stage only, Black race (subdistribution hazard ratio [SHR], 1.30; 95% CI, 1.17-1.44; P < .001) and Latinx ethnicity (SHR, 1.58; 95% CI, 1.37-1.81; P < .001) were associated with an increased 1-year hazard of MALE compared with White race. On adjusted multivariable analysis, MALE disparities persisted for Black/African American race (SHR, 1.12; 95% CI, 1.01-1.25; P = .028) and Latinx/Hispanic ethnicity (SHR, 1.34; 95% CI, 1.16-1.54; P < .001) compared with White race. CONCLUSIONS: Black/African American and Latinx/Hispanic patients had a higher associated hazard of MALE after PVI for CLTI compared with White patients despite an adjustment for WIfI stage at presentation. These results suggest that disease severity at presentation does not account for disparities in outcomes. Further work should focus on better understanding the underlying mechanisms for disparities in historically marginalized racial and ethnic groups presenting with CLTI.

Structured discharge documentation reduces sex-based disparities in statin prescription in vascular surgery patients.

OBJECTIVE: Perioperative statin use has been shown to improve survival in vascular surgery patients. In 2018, the Northern California Vascular Study Group implemented a quality initiative focused on the use of a SmartText in the discharge summary. We hypothesized that structured discharge documentation would decrease sex-based disparities in evidence-based medical therapy. METHODS: A retrospective analysis was conducted using Vascular Quality Initiative eligible cases at a single institution. Open or endovascular procedures in the abdominal aorta or lower extremity arteries from 2016 to 2021 were included. Bivariate analysis identified factors associated with statin use and sex. Multivariate logistic regression was performed with the end point of statin prescription at discharge and aspirin prescription at discharge. An interaction term assessed the differential impact of the initiative on both sexes. Analysis was then stratified by prior aspirin or statin prescription. An interrupted time series analysis was used to evaluate the trend in statin prescription over time. RESULTS: Overall, 866 patients were included, including 292 (34%) female and 574 (66%) male patients. Before implementation, statins were prescribed in 77% of male and 62% of female patients (P < .01). After implementation, there was no statistically significant difference in statin prescription (91% in male vs 92% in female patients, P = .68). Female patients saw a larger improvement in the adjusted odds of statin prescription compared with male patients (odds ratio: 3.1, 95% confidence interval: 1.1-8.6, P = .04). For patients not prescribed a statin preoperatively, female patients again saw an even larger improvement in the odds of being prescribed a statin at discharge (odds ratio: 6.4, 95% confidence interval: 1.8-22.7, P < .01). Interrupted time series analysis demonstrated a sustained improvement in the frequency of prescription for both sexes over time. The unadjusted frequency of aspirin prescription also improved by 3.5% in male patients vs 5.5% in female patients. For patients not prescribed an aspirin preoperatively, we found that the frequency of aspirin prescription significantly improved for both male (19% increase, P = .006) and female (31% increase, P = .001) patients. There was no significant difference in the perioperative outcomes between male and female patients before and after standardized discharge documentation. CONCLUSIONS: A simple, low-cost regional quality improvement initiative eliminated sex-based disparities in statin prescription at a single institution. These findings highlight the meaningful impact of regional quality improvement projects. Future studies should examine the potential for structured discharge documentation to improve patient outcomes and reduce disparities.

The influence of avoidant attachment and perceived support on disclosure about involvement in donor-assisted conception to family and friends.

STUDY QUESTION: Do the attachment-related dimensions Anxiety and Avoidance and perceived partner and social support in recipients and donors influence disclosure to others about their involvement in donor-assisted conception (DAC)? SUMMARY ANSWER: A higher global score on attachment Avoidance was associated with greater non-disclosure about involvement in DAC by participants to relationship-specific others. WHAT IS KNOWN ALREADY: Within the context of DAC, the topic of disclosure has been investigated in terms of the 'if', 'when', and the 'how' to disclose about circumstances of conception. Less focus, however, has been directed to investigating psychological theoretical frameworks that influence disclosure decisions to others, i.e. to whom information is disclosed and to what extent details are transparently revealed about the donor programme. STUDY DESIGN, SIZE, DURATION: The study was of a cross-sectional design and utilized a sample of 301 participants who were, or had been, involved in DAC, and were recruited across states of Australia. An online self-report questionnaire was completed between June 2014 and June 2017. PARTICIPANTS/MATERIALS, SETTING, METHODS: English speaking participants consisting of 209 female recipients and 92 donors (36 sperm; 48 egg; 8 embryo donors). Of the recipients, 104 had successfully conceived children via donated gametes (68 sperm, 23 eggs, 6 embryos, and 7 recipients where both gametes were donated from 2 donors to create the embryo). Participants anonymously completed an online questionnaire consisting of five sections: Demographics, Donor Conception and Disclosure Practices, the Experiences in Close Relationships-Relationships Structure, the Quality of Relationship Inventory, and the Multidimensional Scale of Perceived Social Support. Pearson correlations, independent samples t-tests, Chi-square, and ANOVA were used to explore the association between attachment Anxiety and Avoidance scores and disclosure about involvement in a DAC programme to significant others (i.e. parents, siblings, in-laws, and friends). MAIN RESULTS AND THE ROLE OF CHANCE: Compared to published community cohort data, participants reported lower global scores on attachment Anxiety and Avoidance and high levels of romantic partner and social network support, suggestive of secure relationships in the overall study sample. A higher score on attachment Avoidance was associated with less disclosure to significant others in their social network (i.e. parents, siblings, in-laws, and close friends), even in the presence of strong partner support (partial r = -0.248, P = 0.005). Higher scores on attachment Avoidance were inversely associated with level of perceived partner and social network support (all P < 0.05). Irrespective of attachment scores, more than 90% of all participants agreed that a child born of DAC should be told about mode of conception. LIMITATIONS, REASONS FOR CAUTION: This study utilized a cross-sectional design precluding causal inferences between dimensions of insecure attachment and disclosure practices. Participants were required to self-report on the quality of their relationships with the potential for social desirability respondent bias. The study's self-selecting sample may limit generalization to participants who were dis-inclined to participate. Specifically, respondents who have an Avoidant attachment style, may have elected not to participate in the study. WIDER IMPLICATIONS OF THE FINDINGS: Given the increased use of biotechnology and digital facial recognition enabling self-discovery of the donor and the donor's extended family, non-disclosure about involvement in DAC may have consequences. An 'Avoidant' attachment style is important to assess as a potential risk factor for non-disclosure about involvement in DAC across different relational contexts (e.g. close family members and friends). Fertility counsellors should consider introducing a measure of attachment screening as a pre-emptive psychoeducational strategy during donor implications counselling. This information could be used to offer patients insight into concerns they have about DAC disclosures to key important relationships, providing a target of clinical intervention. STUDY FUNDING/COMPETING INTEREST(S): No external funds were sought for this work. None of the authors have any competing interests to declare. TRIAL REGISTRATION NUMBER: N/A.

A new classification system for the anatomical variations of the human circle of Willis: A systematic review.

The circle of Willis (CoW) is an anastomotic arterial network located on the base of the brain. Studies have shown that it demonstrates considerable anatomical variation in humans. This systematic review aimed to identify and catalogue the described anatomical variations of the CoW in humans to create a new, comprehensive variation classification system. An electronic literature search of five databases identified 5899 studies. A two-phase screening process was performed, and studies underwent quality assessment. A total of 42 studies were included in the review. Data were extracted and circles were reconstructed digitally using graphics software. The classification system contains 82 CoW variations in five continuous groups. Group one contains 24 circles with one or more hypoplastic segments only. Group two contains 11 circles with one or more absent segments only. Group three contains 6 circles with hypoplastic and absent segments only. Group four contains 26 circles with one or more accessory segments. Group five contains 15 circles with other types of anatomical variation. Within each group, circles were subcategorised according to the number or type of segments affected. An original coding system was created to simplify the description of anatomical variations of the CoW. The new classification system provides a comprehensive ontology of the described anatomical variations of the CoW in humans. When used with the coding system, it allows the description and categorisation of recorded and unrecorded variants identified in past and future studies. It is applicable to current clinical practice and the anatomical community, including human anatomy education and research.

The seasonal relationships between household dietary diversity and child growth in a rural Timor-Leste community.

Both child growth and dietary diversity are poor in rural Timor-Leste. The rainy season is associated with food scarcity, yet the association between seasonal scarcity, food diversity, and child growth is underdocumented. This study assesses the relationship between household dietary diversity and children's standardized growth across the 2018 food-scarce (April-May; post-rainy period) and post-harvest (October) seasons in the agricultural community of Natarbora, on the south-coastal plains of Timor-Leste. We conducted household interviews and collected anthropometric data across 98 and 93 households in the post-rainy and post-harvest periods, respectively. Consumed household foods were obtained via 24-h diet recalls and were subsequently categorized into a nine-food-group dietary diversity score (DDS; number of different food groups consumed). The DDS was related to children's standardized short-term growth (z-weight, z-body mass index [BMI] and percent change in weight over the harvest season) via linear mixed models. Across seasons, DDS increased from 3.9 (standard deviation [SD] = 1.0) to 4.3 (SD = 1.4; p < 0.05). In the post-rainy season, children in high DDS households had higher z-weight than those in low DDS households and higher z-BMI than children in medium and low DDS households. In the post-harvest period, household DDS did not predict children's z-weight but predicted z-BMI. Consumption of protein-rich foods, particularly animal-source foods and legumes, in low- and medium-DDS households may be associated with improved child growth. While consuming more animal-source foods in the post-rainy season would be ideal, promoting the consumption of locally grown legumes, such as beans and pulses, may facilitate better nutritional outcomes for more children in rural Timor-Leste.

Understanding value and patient complexity among common inpatient vascular surgery procedures.

OBJECTIVE: Vascular surgery patients are highly complex, second only to patients undergoing cardiac procedures. However, unlike cardiac surgery, work relative value units (wRVU) for vascular surgery were undervalued based on an overall patient complexity score. This study assesses the correlation of patient complexity with wRVUs for the most commonly performed inpatient vascular surgery procedures. METHODS: The 2014 to 2017 National Surgical Quality Improvement Program Participant Use Data Files were queried for inpatient cases performed by vascular surgeons. A previously developed patient complexity score using perioperative domains was calculated based on patient age, American Society of Anesthesiologists class of ≥4, major comorbidities, emergent status, concurrent procedures, additional procedures, hospital length of stay, nonhome discharge, and 30-day major complications, readmissions, and mortality. Procedures were assigned points based on their relative rank and then an overall score was created by summing the total points. An observed to expected ratio (O/E) was calculated using open ruptured abdominal aortic aneurysm repair (rOAAA) as the referent and then applied to an adjusted median wRVU per operative minute. RESULTS: Among 164,370 cases, patient complexity was greatest for rOAAA (complexity score = 128) and the least for carotid endarterectomy (CEA) (complexity score = 29). Patients undergoing rOAAA repair had the greatest proportion of American Society of Anesthesiologists class of ≥IV (84.8%; 95% confidence interval [CI], 82.6%-86.8%), highest mortality (35.5%; 95% CI, 32.8%-38.3%), and major complication rate (87.1%; 95% CI, 85.1%-89.0%). Patients undergoing CEA had the lowest mortality (0.7%; 95% CI, 0.7%-0.8%), major complication rate (8.2%; 95% 95% CI, 8.0%-8.5%), and shortest length of stay (2.7 days; 95% CI, 2.7-2.7). The median wRVU ranged from 10.0 to 42.1 and only weakly correlated with overall complexity (Spearman's ρ = 0.11; P < .01). The median wRVU per operative minute was greatest for thoracic endovascular aortic repair (0.25) and lowest for both axillary-femoral artery bypass (0.12) and open femoral endarterectomy, thromboembolectomy, or reconstruction (0.12). After adjusting for patient complexity, CEA (O/E = 3.8) and transcarotid artery revascularization (O/E = 2.8) had greater than expected O/E. In contrast, lower extremity bypass (O/E = 0.77), lower extremity embolectomy (O/E = 0.79), and open abdominal aortic repair (O/E = 0.80) had a lower than expected O/E. CONCLUSIONS: Patient complexity varies substantially across vascular procedures and is not captured effectively by wRVUs. Increased operative time for open procedures is not adequately accounted for by wRVUs, which may unfairly penalize surgeons who perform complex open operations.

Pandemics, Protests, and Pronouns: The Changing Landscape of Biomedical Visualisation and Education.

Events in early 2020 changed the landscape of education for the foreseeable future, perhaps permanently. Three events had a significant impact; (1) the Coronavirus disease 2019 (COVID-19) pandemic, (2) the death of George Floyd, which resulted in the most recent Black Lives Matter (BLM) protests, and (3) the Twitter storm, the resultant societal fallout and freedom of speech campaigns, following comments made by author JK Rowling which many deemed transphobic. These events had a differential impact on biomedical sciences, when compared to other sectors. COVID-19 resulted in a global lockdown, with higher education institutions closing campuses and moving to online-only delivery. This rapid change required radical shifts in the use of technology, with mass delivery of teaching at short notice. The BLM protests further raised awareness of the inequalities within society, particularly those experienced by Black people and other oppressed groups. As a result, there have been calls for the decolonisation of the curriculum. The implications of these three key events have led institutions to rethink their policies, teaching delivery, assessment, curricula, and physical environments. This chapter considers (1) the implications of a swift change in the primary mode of curriculum delivery within Higher Education to online formats and (2) how recent adverse events have resulted in calls for much-needed changes in visual representations within biomedical sciences. Finally, we consider (3) the role of the hidden curriculum and the potential impact of visual representations in curricula on the delivery of healthcare and the fight against health inequalities, which are often as a result of implicit biases. The year 2020 has proven timely in presenting the opportunity for change, provided through the power of imagery.

Anatomical variations of the circle of Willis and their prevalence, with a focus on the posterior communicating artery: A literature review and meta-analysis.

The circle of Willis is an anastomotic network of arteries surrounding the base of the brain, providing collateral circulation to prevent ischemia. It has, however, long been established that it exhibits considerable anatomical variation when compared to Thomas Willis' originally described circle. This study aimed primarily to determine an accurate prevalence of the variation of the circle of Willis in the general population and the prevalence of common posterior communicating artery variations. Additional aims were to explain why such a wide range of reported variations exist, and whether different types of studies report significantly different prevalence of variation. A comprehensive literature search identified 764 papers. A three-phase screening process was undertaken, involving a critical analysis of papers, and a total of 33 papers were selected for analysis and literature review. A descriptive statistics test with bootstrap was performed to estimate the average prevalence of variations. The estimated prevalence of general variation, unilateral, and bilateral posterior communicating artery hypoplasia or aplasia was 68.22 ± 14.32%, 19.45 ± 8.63%, and 22.83 ± 14.58%, respectively. Over half of the population exhibit a circle of Willis with some form of variation. To provide a more accurate estimation for the prevalence of variations, a universal classification system needs to be established, collating all the work from high-quality studies, to provide a comprehensive database of the circle's variations. Knowing the prevalence of variations and how they can impact neurosurgical approaches or patterns of ischemic pathology can be crucial in providing effective patient care.

Determination of online quenching efficiency for an automated cellular microfluidic metabolomic platform using mass spectrometry based ATP degradation analysis.

As microfluidic cell culture progresses, the need for robust and reproducible intracellular analyses grows. In particular, intracellular metabolites are subject to perturbation and degradation during the lysing process. The reliability of intracellular metabolomic analysis in microfluidic devices depends on the preservation of metabolite integrity during sample preparation and storage. Described here is a novel automated microfluidic system exhibiting the necessary rapid cellular lysis and quenching of enzymatic activity. Quenching efficiency was assessed using a novel ratiometric MALDI-MS-based assay of exogenous isotopic adenosine triphosphate (ATP) hydrolysis to isotopic adenosine diphosphate (ADP) as a marker of metabolite degradation. The lysis system of the microfluidic device was enhanced using a Peltier cooler to chill the lysate and quench aberrant enzymatic activity. Parameter optimization (flow rate, collection time, and temperature control) improved the endogenous and exogenous ADP/ATP ratios by 44.9% and 39.8% respectively consistent with traditional quenching techniques. The effects of chilling/quenching on metabolism were evaluated resulting in over 500 significant features compared to non-chilled from untargeted capillary LC-MS metabolomic analyses. These include increased levels of tryptophan, histidine, and pyruvate as well as decreased levels in UDP-N-acetylglucosamine. The results illustrate the need for both rapid lysis and quenching in microfluidic cell culture platforms. Graphical abstract.

Determinants of objectively measured physical activity in rural East Timorese children.

OBJECTIVES: The human juvenile period evolved as a period of learning and physical development in a family environment that subsidizes the costs of these processes. Children allocate energy to physical activity, maintenance, and growth. How energy is allocated has consequences for adult body size and other life-history traits. In subsistence agriculture populations, where child contributions to the household economy are common and energy availability is low, trade-offs in energy expenditure between activity and growth may help explain poor growth. METHODS: Using accelerometry, we measured physical activity over 2 years in 88 free-living children aged 5-19 years in two ecologically varying communities in rural Timor-Leste. We model characteristics related to variation in activity, and subsequently, activity is modeled against growth, illness, and aspects of household and local ecology using linear mixed models. RESULTS: Physical activity in Timorese children is characterized by high levels of moderate ( x ¯ = 8.8 h/day), no sustained vigorous, and little sedentary activity ( x ¯ = 4.6 h/day). Children in the mountainous community show a slight trade-off between activity and growth (P = .077). Males down-regulate both growth and activity relative to females. Variation in household characteristics does not predict child activity. Both activity and growth are lower in the mountainous community than in the flat, coastal community. CONCLUSIONS: Household demands on child behavior may constrain children's ability to moderate activity relative to nutritional status. Activity in this population is high relative to other subsistence populations, possibly because children face the dual pressures of contributing to household subsistence and attending school.

Growth curves and the international standard: How children's growth reflects challenging conditions in rural Timor-Leste.

OBJECTIVES: Population-specific growth references are important in understanding local growth variation, especially in developing countries where child growth is poor and the need for effective health interventions is high. In this article, we use mixed longitudinal data to calculate the first growth curves for rural East Timorese children to identify where, during development, deviation from the international standards occurs. MATERIALS AND METHODS: Over an eight-year period, 1,245 children from two ecologically distinct rural areas of Timor-Leste were measured a total of 4,904 times. We compared growth to the World Health Organization (WHO) standards using z-scores, and modeled height and weight velocity using the SuperImposition by Translation And Rotation (SITAR) method. Using the Generalized Additive Model for Location, Scale and Shape (GAMLSS) method, we created the first growth curves for rural Timorese children for height, weight and body mass index (BMI). RESULTS: Relative to the WHO standards, children show early-life growth faltering, and stunting throughout childhood and adolescence. The median height and weight for this population tracks below the WHO fifth centile. Males have poorer growth than females in both z-BMI (p = .001) and z-height-for-age (p = .018) and, unlike females, continue to grow into adulthood. DISCUSSION: This is the most comprehensive investigation to date of rural Timorese children's growth, and the growth curves created may potentially be used to identify future secular trends in growth as the country develops. We show significant deviation from the international standard that becomes most pronounced at adolescence, similar to the growth of other Asian populations. Males and females show different growth responses to challenging conditions in this population.

Household resources and seasonal patterns of child growth in rural Timor-Leste.

OBJECTIVES: This study aimed to determine through detailed contextual investigation the effects of seasonal resource shortages, and household and individual level differences, on child growth in rural Timor-Leste. We compared trends in growth across two rural Timorese villages with different ecologies. METHODS: Heads of 104 households in Natarbora, Timor-Leste, were interviewed and resource levels assessed during the food shortage season. In these households, 337 children were measured for height, weight, and mid-upper arm circumference. World Health Organization standardized measures were calculated and compared with dry season measurements. Using hierarchical linear models, child growth was related to household resource levels. Results were then compared to data from rural mountainous Ossu, Timor-Leste. RESULTS: z BMI declined over the wet season when food resources were scarce compared with the dry season (P < .001). Both age and sex were strong predictors of child growth, with older children having worse z height-for-age (P = .001) and z weight-for-age (P < .001) and boys shorter for age than girls (P = .049). Children were taller in households with modern flushable toilets (P = .005). Agricultural strategies such as crop diversity and land cultivation were linked to child growth. Results parallel findings from Ossu on the effects of season, child age and sex, but not household level socioeconomic differences. CONCLUSIONS: Results highlight the importance of subsistence-based resource stabilization and of early intervention to prevent growth faltering. Predicting growth using ecological models requires small-scale investigation, as variation exists among rural areas within an ecologically and culturally diverse country.

Obesity and psychological wellbeing in patients undergoing fertility treatment.

Obesity negatively affects reproductive functioning and psychological wellbeing. Distress experienced by infertile women with elevated body mass index (BMI) was investigated. Infertile women (n = 403) were stratified according to World Health Organization (2000) BMI categories (normal, overweight and obese) and infertility category (polycystic ovary syndrome [PCOS] or non-PCOS). Participants anonymously completed a Demographics Questionnaire, International Physical Activity Questionnaire, Depression, Anxiety and Stress Scale, Positive and Negative Affect Schedule, Rosenberg Self Esteem Scale, Fertility Problem Inventory, Clinical Perfectionism Questionnaire and the Eating Disorder Examination Questionnaire. Women in the obese BMI group were no more psychologically vulnerable to general mood (depression, anxiety and stress) or fertility-specific distress than normal or overweight BMI women. Independent of their PCOS status, obese women reported more frequent episodes of binge eating, shape concerns and low self-esteem symptoms associated with disordered eating. Women with PCOS had elevated shape concerns and anxiety independent of their BMI category compared with women who did not have PCOS. Obese infertile women presenting with the characteristics of binge eating, low self-esteem and body shape concerns may represent a vulnerable subgroup that could benefit from accessing targeted psychological interventions as do women with PCOS who have body shape concerns.

Automated sample preparation in a microfluidic culture device for cellular metabolomics.

Sample pretreatment in conventional cellular metabolomics entails rigorous lysis and extraction steps which increase the duration as well as limit the consistency of these experiments. We report a biomimetic cell culture microfluidic device (MFD) which is coupled with an automated system for rapid, reproducible cell lysis using a combination of electrical and chemical mechanisms. In-channel microelectrodes were created using facile fabrication methods, enabling the application of electric fields up to 1000 V cm(-1). Using this platform, average lysing times were 7.12 s and 3.03 s for chips with no electric fields and electric fields above 200 V cm(-1), respectively. Overall, the electroporation MFDs yielded a ∼10-fold improvement in lysing time over standard chemical approaches. Detection of multiple intracellular nucleotides and energy metabolites in MFD lysates was demonstrated using two different MS platforms. This work will allow for the integrated culture, automated lysis, and metabolic analysis of cells in an MFD which doubles as a biomimetic model of the vasculature.

Disordered eating attitudes and exercise in women undergoing fertility treatment.

BACKGROUND: Previous research has found a higher lifetime prevalence of eating disorders in women undergoing fertility treatment. AIMS: This study aimed to gauge the prevalence of eating disorders in women attending a fertility clinic and to compare current disordered eating attitudes and exercise amongst different infertility categories. MATERIALS AND METHODS: Three hundred and eighty-five women were grouped according to infertility diagnosis: male factor, unexplained infertility, ovulatory disorders, polycystic ovarian syndrome (PCOS) and heterogeneous causations. Participants anonymously completed the Eating Disorder Examination Questionnaire (EDE-Q), the International Physical Activity Questionnaire (IPAQ) and a Demographics questionnaire. RESULTS: The lifetime history of self-reported eating disorders for women in the sample was similar to Australian community rates. Profile analysis revealed on the EDE-Q that women with ovulatory disorders were not significantly different from women with PCOS; however, they were significantly different to women with other infertility diagnoses (all P < 0.05), suggesting increased vulnerability to disordered eating. There were no between-group differences in exercise quantity (IPAQ, P = 0.625) although women with ovulatory disorders and PCOS had a significantly higher risk of engaging in compulsive, 'driven' exercise (OR = 6.98, CI = 1.39, 34.90, P = 0.018) as a means to control weight or shape. CONCLUSIONS: Contrary to previous research, our findings do not confirm a greater lifetime prevalence rate of eating disorders in women attending an infertility clinic. This study does highlight the importance of screening women with forms of an ovulatory disorder for features of disordered eating attitudes and driven exercise behaviours.

Poor Sperm Chromatin Condensation Is Associated with Cryopreservation-Induced DNA Fragmentation and Cell Death in Human Spermatozoa.

Background/Objectives: Semen cryopreservation is routinely performed in fertility clinics for a variety of reasons, including fertility preservation and storage of donor sperm, yet the freeze-thaw process leads to cellular damage via ice crystal formation, osmotic shock, and supraphysiological levels of oxidative stress. Sperm resistance to damage during the freeze-thaw process varies widely, yet the intrinsic factors associated with sperm cryotolerance are largely unknown. The study aimed to investigate whether poor chromatin condensation renders sperm vulnerable to DNA fragmentation and cell death induced by the freeze-thaw process. Methods: Participants (n = 51) from the general community who met the inclusion criteria collected a semen sample after 3-8 days of abstinence. Neat semen samples underwent traditional semen analysis, aniline blue (AB)-eosin staining for chromatin condensation, the terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) assay for DNA fragmentation, and the Annexin V assay for apoptosis/necrosis, prior to being cryopreserved using the liquid nitrogen vapour method and stored at -196 °C. Stored samples were later thawed at room temperature and processed using density gradient centrifugation. Motile sperm concentration, DNA fragmentation and apoptosis/necrosis were analysed in post-thaw samples. Results: As indicated by a significant interaction effect in linear mixed models, an increased proportion of AB-positive sperm in the pre-freeze sample exacerbated the adverse effect of freezing on sperm DNA fragmentation (p = 0.004), late apoptosis (p = 0.007), and necrosis (p = 0.007). AB-staining was positively correlated with all three parameters in the post-thaw sample (all rs ≥ 0.424, all p < 0.01) and remained significant after adjusting for neat sperm concentration (all partial rs ≥ 0.493, all p < 0.01). Similarly, AB-staining was significantly correlated with the percentage point change in sperm DNA fragmentation (rs = 0.366, p = 0.014) and necrosis (rs = 0.403, p = 0.009), both of which remained significant after adjusting for neat sperm concentration (both partial rs ≥ 0.404, both p < 0.01), and borderline significantly correlated with percentage point change in late apoptosis (rs = 0.307, p = 0.051). Conclusions: Sperm with poorly condensed chromatin may be more susceptible to cellular damage during the freeze-thaw process, independent of pre-freeze sperm concentration. These findings may help to explain the intrinsic variation in sperm resistance to cryodamage within and between individuals that is poorly understood.

Multi-omics analysis reveals COVID-19 vaccine induced attenuation of inflammatory responses during breakthrough disease.

The immune mechanisms mediating COVID-19 vaccine attenuation of COVID-19 remain undescribed. We conducted comprehensive analyses detailing immune responses to SARS-CoV-2 virus in blood post-vaccination with ChAdOx1 nCoV-19 or a placebo. Samples from randomised placebo-controlled trials (NCT04324606 and NCT04400838) were taken at baseline, onset of COVID-19-like symptoms, and 7 days later, confirming COVID-19 using nucleic amplification test (NAAT test) via real-time PCR (RT-PCR). Serum cytokines were measured with multiplexed immunoassays. The transcriptome was analysed with long, short and small RNA sequencing. We found attenuation of RNA inflammatory signatures in ChAdOx1 nCoV-19 compared with placebo vaccinees and reduced levels of serum proteins associated with COVID-19 severity. KREMEN1, a putative alternative SARS-CoV-2 receptor, was downregulated in placebo compared with ChAdOx1 nCoV-19 vaccinees. Vaccination ameliorates reductions in cell counts across leukocyte populations and platelets noted at COVID-19 onset, without inducing potentially deleterious Th2-skewed immune responses. Multi-omics integration links a global reduction in miRNA expression at COVID-19 onset to increased pro-inflammatory responses at the mRNA level. This study reveals insights into the role of COVID-19 vaccines in mitigating disease severity by abrogating pro-inflammatory responses associated with severe COVID-19, affirming vaccine-mediated benefit in breakthrough infection, and highlighting the importance of clinically relevant endpoints in vaccine evaluation.

Male age negatively influences clinical pregnancy rate in women younger than 40 years undergoing donor insemination cycles.

The influence of paternal age on clinical pregnancy was examined within younger patients undergoing donor insemination (DI) cycles in Western Australia. A retrospective analysis of 2142 DI cycles was carried out, including only women <40 years with no history of fertility problems. Logistic regression analysis was used to relate donor age to clinical pregnancy rate. Proportional hazards regression analysis was used to relate male age to time to pregnancy. Mediation analysis was conducted to adjust for influence of sperm concentration and motility. Analysis was controlled for female age, luteal-phase progesterone support and insemination year. There was no effect of female age on pregnancy rate or time to pregnancy. Older males (age ≥ 45 years) were significantly associated with a reduced pregnancy rate and longer time to pregnancy. Mediation analysis indicated that sperm concentration and motility did not fully account for the effect of male age on pregnancy. This study reveals an adverse effect of male age on clinical pregnancy in women <40 years of age that is not solely mediated by decreased sperm concentration or motility. This has implications for recruitment systems that attract older donors such as open-identity systems, which may benefit from actively recruiting younger donors.