Determinants of mortality status and population attributable risk fractions of the North West Province, South African site of the international PURE study.

BACKGROUND: Mortality data and comparative risk assessments from sub-Saharan Africa are limited. There is an urgent need for high quality population health surveys to be conducted, to improve the national health surveillance system. Our aim was to perform a comparative risk assesment and report on the mortality status and cause of death data of participants from a South African site of the international Prospective Urban Rural Epidemiology study. METHODS: 1 921 Black participants were included, with a median observational time of 13 years resulting in 21 525 person-years. We performed a comparative risk assessment considering four health status domains: locality (rural vs. urban), socio-economic status (SES) (education and employment), lifestyle factors (physical activity, smoking and alcohol consumption) and prevalent diseases (human immunodeficiency virus (HIV), type 2 diabetes mellitus and hypertension). Next, population-attributable fractions (PAFs) were calculated to determine the mortality risk attributable to modifiable determinants. RESULTS: 577 all-cause deaths occurred. Infectious diseases (28.1% of all deaths) were the most frequent cause of death, followed by cardiovascular disease (CVD) (22.4%), respiratory diseases (11.6%) and cancer (11.1%). The three main contributors to all-cause mortality were HIV infection, high SES and being underweight. HIV infection and underweight were the main contributors to infectious disease mortality and hypertension, the urban environment, and physical inactivity to CVD mortality. HIV had the highest PAF, followed by physical inactivity, alcohol and tobacco use and hypertension (for CVD mortality). CONCLUSION: This African population suffers from a quadruple burden of disease. Urban locality, high SES, prevalent disease (HIV and hypertension) and lifestyle factors (physical inactivity, tobacco and alcohol use) all contributed in varying degrees to all-cause and cause-specific mortalities. Our data confirm the public health importance of addressing HIV and hypertension, but also highlights the importance of physical inactivity, tobacco use and alcohol consumption as focal points for public health strategies to produce the most efficient mortality reduction outcomes.

Painted flowers: Eluta generates pigment patterning in Antirrhinum.

In the early 1900s, Erwin Baur established Antirrhinum majus as a model system, identifying and characterising numerous flower colour variants. This included Picturatum/Eluta, which restricts the accumulation of magenta anthocyanin pigments, forming bullseye markings on the flower face. We identified the gene underlying the Eluta locus by transposon-tagging, using an Antirrhinum line that spontaneously lost the nonsuppressive el phenotype. A candidate MYB repressor gene at this locus contained a CACTA transposable element. We subsequently identified plants where this element excised, reverting to a suppressive Eluta phenotype. El alleles inhibit expression of anthocyanin biosynthetic genes, confirming it to be a regulatory locus. The modes of action of Eluta were investigated by generating stable transgenic tobacco lines, biolistic transformation of Antirrhinum petals and promoter activation/repression assays. Eluta competes with MYB activators for promoter cis-elements, and also by titrating essential cofactors (bHLH proteins) to reduce transcription of target genes. Eluta restricts the pigmentation established by the R2R3-MYB factors, Rosea and Venosa, with the greatest repression on those parts of the petals where Eluta is most highly expressed. Baur questioned the origin of heredity units determining flower colour variation in cultivated A. majus. Our findings support introgression from wild species into cultivated varieties.

Assessment of non-communicable disease risk factors, functional performance, and health-related quality of life in adults: a comparative analysis in low-resourced urban and rural areas of South Africa.

BACKGROUND: Globally, disparities between non-communicable disease (NCD) risk factors, functional performance, and health-related quality of life (HRQoL) exist in people living in rural and low-resourced urban settings. Evidence of these health differences determined with objective NCD risk factors and functional performance measurements in South Africa, is scarce. Therefore, the study aimed to determine the differences in NCD risk factors, functional performance and HRQoL between rural and low-resourced urban areas. METHODS: The study recruited 311 adults (35-80 years) presenting with at least one NCD risk factor from low-resourced urban- (n = 183) and rural (n = 128) communities. Objective measurements of physical activity (PA) by means of combined heart rate and accelerometery, body composition employing skinfolds, peripheral lipid and glucose concentrations, blood pressure, functional performance indicators (handgrip, single leg stand, sit-to-stand, timed-up-and-go speed, predicted peak VO2 max); and HRQoL were measured according to standard procedures. Independent t-tests, Mann-Whitney U, and chi-square tests were performed to determine differences between the variables of low-resourced urban and rural settings. RESULTS: The participants from the low-resourced urban setting were significantly older than the rural residents (59.1 ± 10.7 years vs. 52.8 ± 11.3 years; p = 0.001). NCD risk factors were significantly more prevalent in the low-resourced urban participants compared to rural participants, in particular for elevated systolic (85.8% vs. 62.5%; p = 0.001), and diastolic blood pressure (88.5% vs. 65.6%; p = 0.001), physical inactivity (95.9% vs. 87.7%; p = 0.026), increased cholesterol concentrations (22.1% vs. 8.7%; p = 0.002), and increased waist circumference (61.9% vs. 49.2%; p = 0.027). Low-resourced urban residents presented with a higher average body fat percentage (27.69% ± 7.65% vs. 12.23% ± 4.67%; p < 0.001), and lower moderate to vigorous PA levels (37.19 ± 49.55 [95% CI = 29.12-45.27] vs. 62.92 ± 60.43 min/week [95% CI = 47.95-77.90]; p = 0.003) compared to rural residents. Rural residents showed significantly better functional performance, including peak VO2 (23.99 ± 9.89 vs. 16.95 ± 7.64 ml/min/kg; p = 0.001) and single leg stand (right leg: 44.96 ± 18.47 vs. 20.87 ± 19.18 s; p = 0.001) as well as higher HRQoL for the physical (51.06 ± 8.14% vs. 45.62 ± 11.13%; p < 0.001) and mental (54.75 ± 8.24% vs. 48.91 ± 12.27%; p < 0.001) component scores compared to participants from the low-resourced urban areas. CONCLUSION: NCD risk factors, functional performance, and HRQoL significantly differ in rural communities compared to low-resourced urban communities in South Africa. Urban areas' most prevalent risk factors were elevated blood pressure, physical inactivity, and increased waist circumference. Participants from rural areas demonstrated significantly better functional performance, such as fitness and balance. HRQoL was better in rural settings than in urban settings. Future intervention programmes should be tailored for specific settings.

Relationships between Iron Status and Selected Physical Fitness Components of South African Adolescents: The PAHL-Study.

Poor iron status is detrimental to physical and cognitive performance in adolescents. Due to the limited studies investigating the association between iron status and physical fitness components in adolescents from low- and middle-income countries, we aimed to determine the association of iron status with selected physical fitness components in South African adolescents. A cross-sectional study design, including 178 adolescents (102 girls and 76 boys) from the Physical Activity and Health Longitudinal Study (PAHLS), was followed. Height and weight were measured to calculate the body mass index (BMI). Subsequently, WHO BMI-for-age-specific categorised body fatness. Cardiorespiratory fitness was determined with a 20-m shuttle run test (V˙O2max), and lower-body explosive power by the standing broad jump (SBJ). Fasting haemoglobin (Hb) and ferritin were analysed from blood samples. Correlation analyses determine the association between iron status, explosive power and cardiorespiratory fitness. Of the 178 participants, 18.5% (n = 33) had low Hb, and 14% (n = 25) iron deficiency without anaemia. Significant positive correlations were found between the selected physical fitness components, ferritin, and Hb. In boys, a positive association was found between Hb and SBJ (r = 0.30, p = 0.006), whilst in girls, positive associations were found between ferritin (r = 0.25, p = 0.04) and SBJ, and Hb with both SBJ (r = 0.21, p = 0.03) and V˙O2max (r = 0.32, p = 0.001). Hb concentration remained associated with V˙O2max and SBJ in girls after adjustment for age, whilst in boys, Hb concentration was associated with SBJ. Higher iron status in South African adolescents is associated with higher lower-limb explosive power and cardiorespiratory fitness. We suggest monitoring of haematological parameters, and interventions to improve the iron status of South African adolescents.

Screening and identification of fetal alcohol spectrum disorder in criminal legal settings: A realist review.

BACKGROUND: Screening for fetal alcohol spectrum disorder (FASD) has been identified as a promising approach to improve recognition, understanding and effective response to the unique needs of those with FASD in criminal legal settings. However, to date, there has been limited synthesis of relevant screening tools, indicators, or implementation considerations in this context. AIMS: The present review aimed to synthesise evidence and develop a conceptual framework for understanding how, when, why, for whom and by whom FASD screening tools, items and/or indicators and characteristics serve to accurately identify people with FASD in criminal legal contexts, with consideration of individual and system needs relevant to effective implementation and response. METHODS: A preregistered search was conducted using a modified realist review framework for both peer-reviewed articles and grey literature. Included sources were available in English, which focused on individuals with prenatal alcohol exposure and/or FASD with criminal legal involvement and offered new empirical evidence. Sources were reviewed using the Quality Control Tool for Screening Titles and Abstracts by Second Reviewer framework, extracted using a structured coding form and narratively synthesised. RESULTS: The search yielded 52 sources, 11 FASD screening tools designed for or applied in criminal legal settings and 38 potential FASD indicators or characteristics relevant to identifying people who may have FASD in criminal legal settings, organised into six conceptually related domains. There was limited evidence supporting the psychometric properties of screening tools across populations or settings, though growing evidence highlights the promise of some instruments. Although few studies characterised potential considerations to be made when implementing a screening tool or approach, both system and individual level needs related to recognising and effectively responding to FASD in criminal legal contexts were identified, and findings revealed strong support among legal and clinical professionals regarding the need for FASD screening in these settings. CONCLUSIONS: Findings of this review can be used to inform the development, selection, implementation and evaluation of FASD screening tools in criminal legal settings and underscore a continued need for enhanced resources, policy and cross-sectoral response to better support the needs of people with FASD in the criminal legal contexts.

Isolated Fragments of Intact Microvessels: Tissue Vascularization, Modeling, and Therapeutics.

The microvasculature is integral to nearly every tissue in the body, providing not only perfusion to and from the tissue, but also homing sites for immune cells, cellular niches for tissue dynamics, and cooperative interactions with other tissue elements. As a microtissue itself, the microvasculature is a composite of multiple cell types exquisitely organized into structures (individual vessel segments and extensive vessel networks) capable of considerable dynamics and plasticity. Consequently, it has been challenging to include a functional microvasculature in assembled or fabricated tissues. Isolated fragments of intact microvessels, which retain the cellular composition and structures of native microvessels, are proving effective in a variety of vascularization applications including tissue in vitro disease modeling, vascular biology, mechanistic discovery, and tissue prevascularization in regenerative therapeutics and grafting. In this review, we will discuss the importance of recapitulating native tissue biology and the successful vascularization applications of isolated microvessels.

Strategies of Modelling Incident Outcomes Using Cox Regression to Estimate the Population Attributable Risk.

When the Cox model is applied, some recommendations about the choice of the time metric and the model's structure are often disregarded along with the proportionality of risk assumption. Moreover, most of the published studies fail to frame the real impact of a risk factor in the target population. Our aim was to show how modelling strategies affected Cox model assumptions. Furthermore, we showed how the Cox modelling strategies affected the population attributable risk (PAR). Our work is based on data collected in the North-West Province, one of the two PURE study centres in South Africa. The Cox model was used to estimate the hazard ratio (HR) of mortality for all causes in relation to smoking, alcohol use, physical inactivity, and hypertension. Firstly, we used a Cox model with time to event as the underlying time variable. Secondly, we used a Cox model with age to event as the underlying time variable. Finally, the second model was implemented with age classes and sex as strata variables. Mutually adjusted models were also investigated. A statistical test to the multiplicative interaction term the exposures and the log transformed time to event metric was used to assess the proportionality of risk assumption. The model's fitting was investigated by means of the Akaike Information Criteria (AIC). Models with age as the underlying time variable with age and sex as strata variables had enhanced validity of the risk proportionality assumption and better fitting. The PAR for a specific modifiable risk factor can be defined more accurately in mutually adjusted models allowing better public health decisions. This is not necessarily true when correlated modifiable risk factors are considered.

Vascularized Tissue Organoids.

Tissue organoids hold enormous potential as tools for a variety of applications, including disease modeling and drug screening. To effectively mimic the native tissue environment, it is critical to integrate a microvasculature with the parenchyma and stroma. In addition to providing a means to physiologically perfuse the organoids, the microvasculature also contributes to the cellular dynamics of the tissue model via the cells of the perivascular niche, thereby further modulating tissue function. In this review, we discuss current and developing strategies for vascularizing organoids, consider tissue-specific vascularization approaches, discuss the importance of perfusion, and provide perspectives on the state of the field.

Relationships between physical activity, body mass index, waist circumference and handgrip strength amongst adults from the North West province, South Africa: The PURE study.

BACKGROUND:  Handgrip strength (HGS) serves as a proxy for the functional ability and its association with body composition (BC) and physical activity (PA) in South African adults are less clear. AIM:  We investigated the relationships between PA, body composition and HGS amongst adults. SETTING:  Rural and urban population from North West Province, South Africa. METHODS:  A cross-sectional study design was performed on 688 (198 men; 490 women) adults aged 35-70 years from the 2015 measurement wave of the Prospective Urban and Rural Epidemiological (PURE) study from the North West province of South Africa. The International Physical Activity Questionnaire-Short Form (IPAQ-SF) assessed and a dynamometer determined HGS in kilogram. Body mass index (BMI) and waist circumference were used as measures of body composition. Spearman correlation coefficients determined the relationship between PA, BMI and HGS. RESULTS:  In the study, 22% and 26%, respectively, were overweight and obese with women being more overweight and obese compared to men. Sixty percent of the participants met the recommended 150 min or more of moderate to vigorous PA (MVPA) per week. Handgrip strength of the participants in the study was weaker than the published norms. Handgrip strength significantly (p  0.05) differed by age. A significant positive association was found between HGS and BMI. Age negatively (r = -0.12; p = 0.001) correlated with MVPA per week. CONCLUSION:  High prevalence of overweight and obesity exists in the current adults' sample. It was also evident that poor upper limb muscle strength and MVPA were negatively associated with ageing. Given the health implications of poor strength indicators, PA intervention programmes, comprised of strength activities, for an adult population are urgently recommended.

Point-of-use, automated fabrication of a 3D human liver model supplemented with human adipose microvessels.

Advanced in vitro tissue models better reflect healthy and disease tissue conditions in the body. However, complex tissue models are often manufactured using custom solutions and can be challenging to manufacture to scale. Here, we describe the automated fabrication of a cell-dense, thick (≤ 1 cm), human vascularized liver tissue model using a robotic biomanufacturing platform and off-the-shelf components to build, culture, and sample liver tissues hands-free without compromising tissue health or function. Fabrication of the tissue involved 3D bioprinting and incorporation of primary human hepatocytes, primary human non-parenchymal cells, and isolated fragments of intact human microvessels as vascular precursors. No differences were observed in select assessments of the liver tissues fabricated by hand or via automation. Furthermore, constant media exchange, via perfusion, improved urea output and elevated tissue metabolism. Interestingly, inclusion of adipose-derived human microvessels enhanced functional gene expression, including an enhanced response to a drug challenge. Our results describe the fabrication of a thick liver tissue environment useful for a variety of applications including liver disease modeling, infectious agent studies, and cancer investigations. We expect the automated fabrication of the vascularized liver tissue, at the point of use and using off-the-shelf platforms, eases fabrication of the complex model and increases its utility.

Methods for vascularization and perfusion of tissue organoids.

Tissue organoids or "mini organs" can be invaluable tools for understanding health and disease biology, modeling tissue dynamics, or screening potential drug candidates. Effective vascularization of these models is critical for truly representing the in vivo tissue environment. Not only is the formation of a vascular network, and ultimately a microcirculation, essential for proper distribution and exchange of oxygen and nutrients throughout larger organoids, but vascular cells dynamically communicate with other cells to modulate overall tissue behavior. Additionally, interstitial fluid flow, mediated by a perfused microvasculature, can have profound influences on tissue biology. Thus, a truly functionally and biologically relevant organoid requires a vasculature. Here, we review existing strategies for fabricating and incorporating vascular elements and perfusion within tissue organoids.

A Biofabrication Strategy for a Custom-Shaped, Non-Synthetic Bone Graft Precursor with a Prevascularized Tissue Shell.

Critical-sized defects of irregular bones requiring bone grafting, such as in craniofacial reconstruction, are particularly challenging to repair. With bone-grafting procedures growing in number annually, there is a reciprocal growing interest in bone graft substitutes to meet the demand. Autogenous osteo(myo)cutaneous grafts harvested from a secondary surgical site are the gold standard for reconstruction but are associated with donor-site morbidity and are in limited supply. We developed a bone graft strategy for irregular bone-involved reconstruction that is customizable to defect geometry and patient anatomy, is free of synthetic materials, is cellularized, and has an outer pre-vascularized tissue layer to enhance engraftment and promote osteogenesis. The graft, comprised of bioprinted human-derived demineralized bone matrix blended with native matrix proteins containing human mesenchymal stromal cells and encased in a simple tissue shell containing isolated, human adipose microvessels, ossifies when implanted in rats. Ossification follows robust vascularization within and around the graft, including the formation of a vascular leash, and develops mechanical strength. These results demonstrate an early feasibility animal study of a biofabrication strategy to manufacture a 3D printed patient-matched, osteoconductive, tissue-banked, bone graft without synthetic materials for use in craniofacial reconstruction. The bone fabrication workflow is designed to be performed within the hospital near the Point of Care.

Exercise Intervention Changes the Perceptions and Knowledge of Non-Communicable Disease Risk Factors among Women from a Low-Resourced Setting.

We employed the Health Belief Model (HBM) as a theoretical lens to explore the influence of an exercise intervention on the perceptions and knowledge of modifiable risk factors for non-communicable diseases (NCDs) among women from a low-resource setting in South Africa. We used a mixed-methods design, gathering qualitative and quantitative data at baseline (n = 95) and again after 12 weeks (n = 55) and 24 weeks (n = 44) of an exercise intervention. Qualitative data consisted of focus group discussions exploring the knowledge and perceptions of modifiable risk factors for NCDs at the three time points. We collected quantitative measurements of modifiable risk factors for NCDs (waist-to-hip ratio, body mass index, blood pressure, peripheral blood glucose, and cholesterol) as well as objective physical activity (PA) data over seven consecutive days. Surveys on coronary heart disease and PA knowledge were conducted at all three time points. Qualitative findings indicated that health exposures and cultural traditions influenced the participant's perceptions about PA and NCDs. Waist circumference significantly decreased at 12 weeks compared to baseline MD = 4.16, p < 0.001. There was significant improvement at 12 weeks, compared to baseline, MD = 0.59, p = 0.009 for PA knowledge, and MD = 0.68, p = 0.003 for heart disease knowledge. There were reductions from baseline to 24 weeks in diastolic blood pressure (MD = 4.97, p = 0.045), waist circumference (MD = 2.85, p = 0.023) and BMI (MD = 0.82, p = 0.004). Significant heart disease knowledge improvements were found at 24 weeks compared to baseline (MD = 0.75, p < 0.001). Supervised exercise positively influenced Black African females' health behaviours by understanding cultural perceptions of modifiable risk factors for NCDs.

The red flesh of kiwifruit is differentially controlled by specific activation-repression systems.

Anthocyanins are visual cues for pollination and seed dispersal. Fruit containing anthocyanins also appeals to consumers due to its appearance and health benefits. In kiwifruit (Actinidia spp.) studies have identified at least two MYB activators of anthocyanin, but their functions in fruit and the mechanisms by which they act are not fully understood. Here, transcriptome and small RNA high-throughput sequencing were used to comprehensively identify contributors to anthocyanin accumulation in kiwifruit. Stable overexpression in vines showed that both 35S::MYB10 and MYB110 can upregulate anthocyanin biosynthesis in Actinidia chinensis fruit, and that MYB10 overexpression resulted in anthocyanin accumulation which was limited to the inner pericarp, suggesting that repressive mechanisms underlie anthocyanin biosynthesis in this species. Furthermore, motifs in the C-terminal region of MYB10/110 were shown to be responsible for the strength of activation of the anthocyanic response. Transient assays showed that both MYB10 and MYB110 were not directly cleaved by miRNAs, but that miR828 and its phased small RNA AcTAS4-D4(-) efficiently targeted MYB110. Other miRNAs were identified, which were differentially expressed between the inner and outer pericarp, and cleavage of SPL13, ARF16, SCL6 and F-box1, all of which are repressors of MYB10, was observed. We conclude that it is the differential expression and subsequent repression of MYB activators that is responsible for variation in anthocyanin accumulation in kiwifruit species.

Back-Carrying in Children Is Related to Lower Limb Development.

Back-carrying of children is a culturally accepted method of transport and safekeeping of babies in many cultures. Developmental consequences related to back-carrying practices have not been directly investigated. This study determined the relationship between frontal and transverse plane lower limb (LL) development, and back-carrying practices, in black Setswana-speaking children. In 691 2- to 9-year-old Setswana-speaking children, the tibiofemoral angle, intermalleolar distance, femoral anteversion angle (AVA) and tibial torsion angle (TTA), were measured to determine LL development. Back-carrying practices were recorded with a questionnaire and Classification and Regression Tree (CART) was used for the analyses. Significant (p < 0.001) relationships, between back-carrying practices and LL development, were discovered. Statistically significant greater genu valgum (F(5, 690) = 7.2, p < 0.001), greater internal TTAs (F(9, 684) = 17.8, p < 0.001), and smaller AVAs (F(13, 685) = 5.1, p < 0.001) were observed in children back-carried more frequently than children back-carried less frequently. There are relationships between back-carrying practices and LL development in both the frontal and transverse plane. However, the genu valgum, internal TTA and smaller AVA noted in more frequently back-carried children is still within normal limits, thus no educational intervention in back-carrying methods or durations is required. Further research should determine the exact back-carrying practice factors (age until which the child is back-carried) impacting lower limb development the greatest.

Lifestyle factors associated with the transition from healthy to unhealthy adiposity among black South African adults over 10 years.

BACKGROUND AND AIMS: Obesity is associated with an increasing prevalence of cardiovascular diseases in Africa, but some obese individuals maintain cardiometabolic health. The aims were to track metabolically healthy overweight or obesity (MHO) over 10 years in African adults and to identify factors associated with a transition to metabolically unhealthy overweight or obesity (MUO). METHODS AND RESULTS: The participants were the South African cohort of the international Prospective Urban and Rural Epidemiological study. From the baseline data of 1937 adults, 649 women and 274 men were followed for 10 years. The combined overweight and obesity prevalence of men (19.2%-23.8%, p = .02) and women (58%-64.7%, p < .001), and the prevalence of the metabolic syndrome in all participants (25.4%-40.2%, p < .001) increased significantly. More than a quarter (26.2%) of the women and 10.9% of men were MHO at baseline, 11.4% of women and 5.1% of men maintained MHO over 10 years, while similar proportions (12.3% of women, 4.7% of men) transitioned to MUO. Female sex, age, and total fat intake were positively associated with a transition to MUO over 10 years, while physical activity was negatively associated with the transition. HIV positive participants were more likely to be MHO at follow-up than their HIV negative counterparts. CONCLUSIONS: One in two black adults with BMI ≥25 kg/m2 maintained MHO over 10 years, while a similar proportion transitioned into MUO. Interventions should focus on lower fat intakes and higher physical activity to prevent the transition to MUO.

Quantifying Vascular Density in Tissue Engineered Constructs Using Machine Learning.

Given the considerable research efforts in understanding and manipulating the vasculature in tissue health and function, making effective measurements of vascular density is critical for a variety of biomedical applications. However, because the vasculature is a heterogeneous collection of vessel segments, arranged in a complex three-dimensional architecture, which is dynamic in form and function, it is difficult to effectively measure. Here, we developed a semi-automated method that leverages machine learning to identify and quantify vascular metrics in an angiogenesis model imaged with different modalities. This software, BioSegment, is designed to make high throughput vascular density measurements of fluorescent or phase contrast images. Furthermore, the rapidity of assessments makes it an ideal tool for incorporation in tissue manufacturing workflows, where engineered tissue constructs may require frequent monitoring, to ensure that vascular growth benchmarks are met.

The Association between Maladaptive Coping and Treatment Motivation in Individuals who are Sexually Attracted to Children Living in the Community.

There is a need for community treatment programs for people who are sexually attracted to children, but individuals report difficulty accessing services. Individuals who are sexually attracted to children (n = 293) completed an online anonymous survey that revealed a significant positive association between maladaptive coping and two factors of the treatment motivation measure. The association between maladaptive coping and treatment motivation was attenuated at higher levels of ego dystonic distress/aversion. Results highlight the importance of targeting stigma toward those who are sexually attracted to children to increase treatment seeking behavior.

Predictors of Vaping Behavior Change in Young Adults Using the Transtheoretical Model: A Multi-Country Study.

Electronic nicotine products remain popular among college students. These products contain heavy metals, carcinogens and the addictive substance nicotine. By understanding where students are in their behavior change can help to determine and focus messages and campaigns. The aim of this study was to assess predictors of the Transtheoretical model in college-aged vaping. This study consisted of an online/Ipad delivery of a voluntary survey to 1249 young adults/college students at 5 universities (International and within the U.S.). Data was analyzed using Stata. Regression analysis was performed to assess predictors of the stages in the Transtheoretical Model of behavior change. Our study found that women tended to be further along in the stages of change as compared to their male counterparts. Additionally, the older students were more likely to be in maintenance stage as compared to the younger students. The students who vaped longer tended to not have advance into any of the stages of change besides precontemplation. Understanding where students are in the stages of change can help to inform behavioral message campaigns enabling more focused targeting of messages and efforts to reduce consumption. Electronic nicotine products are highly prevalent on college campuses, both nationally and internationally. The nicotine is addictive and may result in less of a desire or ability to quit as young adults age.

Discrete bHLH transcription factors play functionally overlapping roles in pigmentation patterning in flowers of Antirrhinum majus.

Floral pigmentation patterning is important for pollinator attraction as well as aesthetic appeal. Patterning of anthocyanin accumulation is frequently associated with variation in activity of the Myb, bHLH and WDR transcription factor complex (MBW) that regulates anthocyanin biosynthesis. Investigation of two classic mutants in Antirrhinum majus, mutabilis and incolorata I, showed they affect a gene encoding a bHLH protein belonging to subclade bHLH-2. The previously characterised gene, Delila, which encodes a bHLH-1 protein, has a bicoloured mutant phenotype, with residual lobe-specific pigmentation conferred by Incolorata I. Both Incolorata I and Delila induce expression of the anthocyanin biosynthetic gene DFR. Rosea 1 (Myb) and WDR1 proteins compete for interaction with Delila, but interact positively to promote Incolorata I activity. Delila positively regulates Incolorata I and WDR1 expression. Hierarchical regulation can explain the bicoloured patterning of delila mutants, through effects on both regulatory gene expression and the activity of promoters of biosynthetic genes like DFR that mediate MBW regulation. bHLH-1 and bHLH-2 proteins contribute to establishing patterns of pigment distribution in A. majus flowers in two ways: through functional redundancy in regulating anthocyanin biosynthetic gene expression, and through differences between the proteins in their ability to regulate genes encoding transcription factors.