NMR-based metabolomics of plasma from dairy calves infected with two primary causal agents of bovine respiratory disease (BRD).

Each year, bovine respiratory disease (BRD) results in significant economic loss in the cattle sector, and novel metabolic profiling for early diagnosis represents a promising tool for developing effective measures for disease management. Here, 1H-nuclear magnetic resonance (1H-NMR) spectra were used to characterize metabolites from blood plasma collected from male dairy calves (n = 10) intentionally infected with two of the main BRD causal agents, bovine respiratory syncytial virus (BRSV) and Mannheimia haemolytica (MH), to generate a well-defined metabolomic profile under controlled conditions. In response to infection, 46 metabolites (BRSV = 32, MH = 33) changed in concentration compared to the uninfected state. Fuel substrates and products exhibited a particularly strong effect, reflecting imbalances that occur during the immune response. Furthermore, 1H-NMR spectra from samples from the uninfected and infected stages were discriminated with an accuracy, sensitivity, and specificity ≥ 95% using chemometrics to model the changes associated with disease, suggesting that metabolic profiles can be used for further development, understanding, and validation of novel diagnostic tools.

Role of Adenylyl Cyclase Type 7 in Functions of BV-2 Microglia.

To assess the role of adenylyl cyclase type 7 (AC7) in microglia's immune function, we generated AC7 gene knockout (AC7 KO) clones from a mouse microglial cell line, BV-2, using the CRISPR-Cas9 gene editing system. The ability of BV-2 cells to generate cAMP and their innate immune functions were examined in the presence or absence of ethanol. The parental BV-2 cells showed robust cAMP production when stimulated with prostaglandin-E1 (PGE1) and ethanol increased cAMP production in a dose-dependent manner. AC7 KO clones of BV-2 cells showed diminished and ethanol-insensitive cAMP production. The phagocytic activity of the parental BV-2 cells was inhibited in the presence of PGE1; AC7 KO BV-2 cells showed lower and PGE1-insensitive phagocytic activity. Innate immune activities of the parental BV-2 cells, including bacterial killing, nitric oxide synthesis, and expression of arginase 1 and interleukin 10 were activated as expected with small effects of ethanol. However, the innate immune activities of AC7 KO cells were either drastically diminished or not detected. The data presented suggest that AC7 has an important role in the innate immune functions of microglial cells. AC7's involvement in ethanol's effects on immune functions remains unclear. Further studies are needed.

Streptococcus pneumoniae metal homeostasis alters cellular metabolism.

Streptococcus pneumoniae colonizes the human nasopharyngeal mucosa and is a leading cause of community-acquired pneumonia, acute otitis media, and bacterial meningitis. Metal ion homeostasis is vital to the survival of this pathogen across diverse biological sites and contributes significantly to colonization and invasive disease. Microarray and qRT-PCR analysis revealed an upregulation of an uncharacterized operon (SP1433-1438) in pneumococci subjected to metal-chelation by N,N,N',N'-tetrakis-(2-pyridylmethyl)ethylenediamine (TPEN). Supplementation of zinc, cobalt, and nickel following TPEN treatment significantly abrogated induction. BLASTP comparisons and protein topology analysis predicted this locus to encode components of ATP binding cassette (ABC) transporters involved in multidrug resistance (SP1434-1435) and energy-coupling factor (ECF) transporters (SP1436-1438). Inductively coupled plasma mass spectrometry (ICP-MS) analysis identified differences in intracellular metal content in a Δ1434-8 mutant strain compared to parental T4R. Further, analysis of the secreted metabolome of WT and Δ1434-8 strains identified significant changes in pneumococcal glycolytic and amino acid metabolic pathways, indicating a shift towards mixed acid fermentation. Additionally, proteomic analysis revealed differentially expressed proteins in the Δ1434-8 mutant strain, with nearly 20% regulated by the global catabolite repressor, CcpA. Based on these findings, we propose that the transporters encoded by SP1433-1438 are involved in regulating the central metabolism of S. pneumoniae and contributing to bacterial survival during metal stress.

Parent recommendations to support physical activity for families with young children: Results of interviews in deprived and affluent communities in South Wales (United Kingdom).

BACKGROUND: Physical inactivity is the fourth leading cause of mortality worldwide. Early childhood is a critical period when healthy behaviours can be instilled for a future active lifestyle. We explored community, societal and environmental factors affecting child and family physical activity and sought parent recommendations to support physical activity in families with young children. METHODS: We interviewed 61 parents expecting a child or with a baby ≤12 months (35 mother and father paired interviews and 26 interviews with mothers only). We purposively sampled families for neighbourhood deprivation status (Townsend Index; 26 affluent; 35 deprived). We conducted thematic analysis of interview transcripts using Bronfenbrenner's socio-ecological framework to guide interpretation. RESULTS: We identified four themes: work family-life balance; spaces for activity; beliefs and attitudes; and physical activity facilitators. We found that parents from deprived neighbourhoods were more likely to be underactive because of a complex web of community, social and personal factors which reduced motivation and hindered opportunity for physical activity. To increase knowledge and opportunity, respondents suggested 'help not tell' messages covering 'why', 'how' and 'where' information about physical activity, and using physical activity to support community engagement and social interaction. CONCLUSIONS: Recommendations from parents highlight effective communication about the importance of early child and family physical activity and improved community access to safe facilities and opportunities. Both parents need to be engaged in designing interventions to support greater physical activity and healthy behaviours which are relevant and achievable in individuals' lives.

Effect of Biochar on Microbial Growth: A Metabolomics and Bacteriological Investigation in E. coli.

Biochar has been proposed as a soil amendment in agricultural applications due to its advantageous adsorptive properties, high porosity, and low cost. These properties allow biochar to retain soil nutrients, yet the effects of biochar on bacterial growth remain poorly understood. To examine how biochar influences microbial metabolism, Escherichia coli was grown in a complex, well-defined media and treated with either biochar or activated carbon. The concentration of metabolites in the media were then quantified at several time points using NMR spectroscopy. Several metabolites were immediately adsorbed by the char, including l-asparagine, l-glutamine, and l-arginine. However, we find that biochar quantitatively adsorbs less of these metabolic precursors when compared to activated carbon. Electron microscopy reveals differences in surface morphology after cell culture, suggesting that Escherichia coli can form biofilms on the surfaces of the biochar. An examination of significant compounds in the tricarboxylic acid cycle and glycolysis reveals that treatment with biochar is less disruptive than activated carbon throughout metabolism. While both biochar and activated carbon slowed growth compared to untreated media, Escherichia coli in biochar-treated media grew more efficiently, as indicated by a longer logarithmic growth phase and a higher final cell density. This work suggests that biochar can serve as a beneficial soil amendment while minimizing the impact on bacterial viability. In addition, the experiments identify a mechanism for biochar's effectiveness in soil conditioning and reveal how biochar can alter specific bacterial metabolic pathways.

Protein Interactions with Nanoparticle Surfaces: Highlighting Solution NMR Techniques.

In the last decade, nanoparticles (NPs) have become a key tool in medicine and biotechnology as drug delivery systems, biosensors and diagnostic devices. The composition and surface chemistry of NPs vary based on the materials used: typically organic polymers, inorganic materials, or lipids. Nanoparticle classes can be further divided into sub-categories depending on the surface modification and functionalization. These surface properties matter when NPs are introduced into a physiological environment, as they will influence how nucleic acids, lipids, and proteins will interact with the NP surface. While small-molecule interactions are easily probed using NMR spectroscopy, studying protein-NP interactions using NMR introduces several challenges. For example, globular proteins may have a perturbed conformation when attached to a foreign surface, and the size of NP-protein conjugates can lead to excessive line broadening. Many of these challenges have been addressed, and NMR spectroscopy is becoming a mature technique for in situ analysis of NP binding behavior. It is therefore not surprising that NMR has been applied to NP systems and has been used to study biomolecules on NP surfaces. Important considerations include corona composition, protein behavior, and ligand architecture. These features are difficult to resolve using classical surface and material characterization strategies, and NMR provides a complementary avenue of characterization. In this review, we examine how solution NMR can be combined with other analytical techniques to investigate protein behavior on NP surfaces.

Genetic and pharmacological inhibition of TTK impairs pancreatic cancer cell line growth by inducing lethal chromosomal instability.

Pancreatic ductal adenocarcinoma, which accounts for the majority of pancreatic cancers, is a lethal disease with few therapeutic options. Genomic profiling of pancreatic ductal adenocarcinoma has identified a complex and heterogeneous landscape. Understanding the molecular characteristics of pancreatic ductal adenocarcinoma will facilitate the identification of potential therapeutic strategies. We analyzed the gene expression profiles of primary tumors from patients compared to normal pancreas and identified high co-overexpression of core components of the spindle assembly checkpoint, including the protein kinase TTK (also known as MPS-1). We found overexpression of TTK protein in a subset of pancreatic ductal adenocarcinoma primary tumors and cell lines. siRNA-mediated depletion or catalytic inhibition of TTK resulted in an aberrant cell cycle profile, multi- and micro-nucleation, induction of apoptosis, and decreased cell proliferation and transformed growth. Selective catalytic inhibition of TTK caused override of the spindle assembly checkpoint-induced cell cycle arrest. Interestingly, we identified ubiquitin specific peptidase 16 (Usp16), an ubiquitin hydrolase, as a phosphorylation substrate of TTK. Usp16 regulates chromosomal condensation and G2/M progression by deubiquitinating histone H2A and polo-like kinase 1. Phosphomimetic mutants of Usp16 show enhanced proteosomal degradation and may prolong the G2/M transition allowing for correction of replication errors. Taken together, our results suggest a critical role for TTK in preventing aneuploidy-induced cell death in pancreatic cancer.

Educational Attainment at Age 10-11 Years Predicts Health Risk Behaviors and Injury Risk During Adolescence.

PURPOSE: To examine the effect of educational attainment in primary school on later adolescent health. METHODS: Education data attainments at age 7 and 11 were linked with (1) primary and secondary care injury consultation/admissions and (2) the Health Behaviour in School-aged Children survey. Cox regression was carried out to examine if attainment in primary school predicts time to injury in adolescence. RESULTS: Pupils that achieve attainment at age 7 but not at age 11 (i.e., declining attainment over time in primary school) are more likely to have an injury during adolescence. These children are also more likely to self-report drinking in adolescence. CONCLUSIONS: Interventions aimed at children with declining attainment in primary school could help to improve adolescent health.

Role of an adenylyl cyclase isoform in ethanol's effect on cAMP regulated gene expression in NIH 3T3 cells.

Previous research has indicated that the cyclic AMP (cAMP) signal transduction system plays an important role in the predisposition to and development of ethanol abuse in humans. Our laboratory has demonstrated that ethanol is capable of enhancing adenylyl cyclase (AC) activity. This effect is AC isoform-specific; type 7 AC (AC7) is most enhanced by ethanol. Therefore, we hypothesized that the expression of a specific AC isoform will play a role on the effect of ethanol on cAMP regulated gene expression. We employed NIH 3T3 cells transfected with AC7 or AC3 as a model system. To evaluate ethanol's effects on cAMP regulated gene expression, a luciferase reporter gene driven by a cAMP inducing artificial promoter was utilized. Stimulation of AC activity leads to an increase in the reporter gene activity. This increase was enhanced in the presence of ethanol in cells expressing AC7, while cells expressing AC3 did not respond to ethanol. cAMP reporter gene expression was increased in the presence of 8-bromo-cAMP; this expression was not enhanced by ethanol. These observations are consistent with our hypothesis. The basal level of CREB phosphorylation was high and did not change by cAMP stimulation or in the presence of ethanol. However, there were significant changes in the TORC3 amount in nuclei depending on stimulation conditions. The results suggest that nuclear translocation of TORC3 plays a more important role than CREB phosphorylation in the observed changes in the cAMP driven reporter gene activity.

Obesity in pregnancy: infant health service utilisation and costs on the NHS.

OBJECTIVE: To estimate the direct healthcare cost of infants born to overweight or obese mothers to the National Health Service in the UK. DESIGN: Retrospective prevalence-based study. SETTING: Combined linked anonymised electronic data sets on a cohort of mother-child pairs enrolled on the Growing Up in Wales: Environments for Healthy Living (EHL) study. Infants were categorised according to maternal early-pregnancy body mass index (BMI): healthy weight mother (18.5≤BMI<25 kg/m(2); n=342), overweight mother (25≤BMI≤29.9 kg/m(2); n=157) and obese mother (BMI≥30; n=110). PARTICIPANTS: 609 singleton pregnancies with available health service records and an antenatal maternal BMI. PRIMARY OUTCOME MEASURE: Total health service utilisation and direct healthcare costs for providing these services in the year 2012-2013. Costs are calculated as cost of the infant (no maternal costs considered) and are related to health service usage from birth to age 1 year. RESULTS: A strong association existed between healthcare usage cost and BMI (p<0.001). Mean total costs were 72% higher among children born to obese mothers (rate ratio (RR) 1.72, 95% CI 1.71 to 1.73) compared with infants born to healthy weight mothers. Higher costings were attributed to a significantly greater number (RR 1.39, 95% CI 1.04 to 1.84) and duration (RR 1.55, 95% CI 1.37 to 1.74) of inpatient visits and a higher number of general practitioner visits (RR 1.10, 95% CI 1.03 to 1.16). Total mean additional resource cost was estimated at £65.13 for infants born to overweight mothers and £1138.11 for infants born to obese mothers, when compared with infants of healthy weight mothers. CONCLUSIONS: Increasingly infants born to mothers with high BMIs consume additional health service resources in the first year of life; this was apparent across inpatient and general practitioner services. Considering both maternal and infant health service use, interventions that cost less than £2310 per person in reducing obesity early pregnancy could be cost-effective.

Classification of accelerometer wear and non-wear events in seconds for monitoring free-living physical activity.

OBJECTIVE: To classify wear and non-wear time of accelerometer data for accurately quantifying physical activity in public health or population level research. DESIGN: A bi-moving-window-based approach was used to combine acceleration and skin temperature data to identify wear and non-wear time events in triaxial accelerometer data that monitor physical activity. SETTING: Local residents in Swansea, Wales, UK. PARTICIPANTS: 50 participants aged under 16 years (n=23) and over 17 years (n=27) were recruited in two phases: phase 1: design of the wear/non-wear algorithm (n=20) and phase 2: validation of the algorithm (n=30). METHODS: Participants wore a triaxial accelerometer (GeneActiv) against the skin surface on the wrist (adults) or ankle (children). Participants kept a diary to record the timings of wear and non-wear and were asked to ensure that events of wear/non-wear last for a minimum of 15 min. RESULTS: The overall sensitivity of the proposed method was 0.94 (95% CI 0.90 to 0.98) and specificity 0.91 (95% CI 0.88 to 0.94). It performed equally well for children compared with adults, and females compared with males. Using surface skin temperature data in combination with acceleration data significantly improved the classification of wear/non-wear time when compared with methods that used acceleration data only (p<0.01). CONCLUSIONS: Using either accelerometer seismic information or temperature information alone is prone to considerable error. Combining both sources of data can give accurate estimates of non-wear periods thus giving better classification of sedentary behaviour. This method can be used in population studies of physical activity in free-living environments.

Parental recommendations for population level interventions to support infant and family dietary choices: a qualitative study from the Growing Up in Wales, Environments for Healthy Living (EHL) study.

BACKGROUND: Childhood obesity presents a challenge to public health. This qualitative study explored the main barriers to dietary choices faced by parents with infants, and the types of interventions and policy level recommendations they would like to see put in place, to promote a healthier food environment. METHODS: 61 semi-structured interviews with prospective parents and parents of infants (61 mothers and 35 fathers) were conducted. Families were selected according to community deprivation levels using the Townsend Deprivation Index to ensure a representative sample from deprived and affluent neighbourhoods. Inductive thematic analysis was used to analyse the data. RESULTS: Parents identified triggers which led to unhealthy dietary choices such as reliance on fast food outlets due to; shift work, lack of access to personal transport, inability to cook, their own childhood dietary experiences, peer pressure and familial relationships. Parents who made healthy dietary choices reported learning cooking skills while at university, attending community cooking classes, having access to quality food provided by church and community organisations or access to Healthy Start vouchers. They called for a reduction in supermarket promotion of unhealthy food and improved access to affordable and high-quality fresh produce in the local area and in supermarkets. There was a strong message to policy makers to work with commercial companies (food manufactures) as they have resources to lower costs and target messages at a diverse population. Provision of targeted advice to fathers, minority ethnic parents, and tailored and practical advice and information on how to purchase, prepare, store and cook food was requested, along with community cookery classes and improved school cookery lessons. CONCLUSIONS: There is a need for parent directed community/population level interventions that aims to reduce socio-ecological barriers to making healthy dietary choices. Parents desired improvements in meals provided in workplaces, schools and hospitals, as well as increased access to healthy foods by increasing local healthy food outlets and reducing unhealthy, fast food outlets. Knowledge and skills could then be enhanced in line with these improvements, with confidence gained around cooking and storing food appropriately.

Physical activity and excess weight in pregnancy have independent and unique effects on delivery and perinatal outcomes.

BACKGROUND: This study examines the effect of low daily physical activity levels and overweight/obesity in pregnancy on delivery and perinatal outcomes. METHODS: A prospective cohort study combining manually collected postnatal notes with anonymised data linkage. A total of 466 women sampled from the Growing Up in Wales: Environments for Healthy Living study. Women completed a questionnaire and were included in the study if they had an available Body mass index (BMI) (collected at 12 weeks gestation from antenatal records) and/or a physical activity score during pregnancy (7-day Actigraph reading). The full statistical model included the following potential confounding factors: maternal age, parity and smoking status. Main outcome measures included induction rates, duration of labour, mode of delivery, infant health and duration of hospital stay. FINDINGS: Mothers with lower physical activity levels were more likely to have an instrumental delivery (including forceps, ventouse and elective and emergency caesarean) in comparison to mothers with higher activity levels (adjusted OR:1.72(95%CI: 1.05 to 2.9)). Overweight/obese mothers were more likely to require an induction (adjusted OR:1.93 (95%CI 1.14 to 3.26), have a macrosomic baby (adjusted OR:1.96 (95%CI 1.08 to 3.56) and a longer hospital stay after delivery (adjusted OR:2.69 (95%CI 1.11 to 6.47). CONCLUSIONS: The type of delivery was associated with maternal physical activity level and not BMI. Perinatal outcomes (large for gestational age only) were determined by maternal BMI.

Obesity in pregnancy: a retrospective prevalence-based study on health service utilisation and costs on the NHS.

OBJECTIVE: To estimate the direct healthcare cost of being overweight or obese throughout pregnancy to the National Health Service in Wales. DESIGN: Retrospective prevalence-based study. SETTING: Combined linked anonymised electronic datasets gathered on a cohort of women enrolled on the Growing Up in Wales: Environments for Healthy Living (EHL) study. Women were categorised into two groups: normal body mass index (BMI; n=260) and overweight/obese (BMI>25; n=224). PARTICIPANTS: 484 singleton pregnancies with available health service records and an antenatal BMI. PRIMARY OUTCOME MEASURE: Total health service utilisation (comprising all general practitioner visits and prescribed medications, inpatient admissions and outpatient visits) and direct healthcare costs for providing these services in the year 2011-2012. Costs are calculated as cost of mother (no infant costs are included) and are related to health service usage throughout pregnancy and 2 months following delivery. RESULTS: There was a strong association between healthcare usage cost and BMI (p<0.001). Adjusting for maternal age, parity, ethnicity and comorbidity, mean total costs were 23% higher among overweight women (rate ratios (RR) 1.23, 95% CI 1.230 to 1.233) and 37% higher among obese women (RR 1.39, 95% CI 1.38 to 1.39) compared with women with normal weight. Adjusting for smoking, consumption of alcohol, or the presence of any comorbidities did not materially affect the results. The total mean cost estimates were £3546.3 for normal weight, £4244.4 for overweight and £4717.64 for obese women. CONCLUSIONS: Increased health service usage and healthcare costs during pregnancy are associated with increasing maternal BMI; this was apparent across all health services considered within this study. Interventions costing less than £1171.34 per person could be cost-effective if they reduce healthcare usage among obese pregnant women to levels equivalent to that of normal weight women.

Mothers' perspectives on the delivery of childhood injury messages: a qualitative study from the growing up in Wales, environments for healthy living study (EHL).

BACKGROUND: Childhood injury is the second leading cause of death for infants aged 1-5 years in the United Kingdom (UK) and most unintentional injuries occur in the home. We explored mothers' knowledge and awareness of child injury prevention and sought to discover mothers' views about the best method of designing interventions to deliver appropriate child safety messages to prevent injury in the home. METHODS: Qualitative study based on 21 semi-structured interviews with prospective mothers and mothers of young children. Mothers were selected according to neighbourhood deprivation status. RESULTS: There was no difference in awareness of safety devices according to mothers' deprivation status. Social networks were important in raising awareness and adherence to child safety advice. Mothers who were recent migrants had not always encountered safety messages or safety equipment commonly used in the UK. Mothers' recommended that safety information should be basic and concise, and include both written and pictorial information and case studies focus on proactive preventive messages. Messages should be delivered both by mass media and suitably trained individuals and be timed to coincide with pregnancy and repeated at age appropriate stages of child development. CONCLUSIONS: The findings suggest that timely childhood injury-related risk messages should be delivered during pregnancy and in line with developmental milestones of the child, through a range of sources including social networks, mass media, face-to-face advice from health professionals and other suitably trained mothers. In addition information on the safe use of home appliances around children and use of child safety equipment should be targeted specifically at those who have recently migrated to the United Kingdom.

Protocol of the baseline assessment for the Environments for Healthy Living (EHL) Wales cohort study.

BACKGROUND: Health is a result of influences operating at multiple levels. For example, inadequate housing, poor educational attainment, and reduced access to health care are clustered together, and are all associated with reduced health. Policies which try to change individual people's behaviour have limited effect when people have little control over their environment. However, structural environmental change and an understanding of the way that influences interact with each other, has the potential to facilitate healthy choices irrespective of personal resources. The aim of Environments for Healthy Living (EHL) is to investigate the impact of gestational and postnatal environments on health, and to examine where structural change can be brought about to optimise health outcomes. The baseline assessment will focus on birth outcomes and maternal and infant health. METHODS/DESIGN: EHL is a longitudinal birth cohort study. We aim to recruit 1000 pregnant women in the period April 2010 to March 2013. We will examine the impact of the gestational environment (maternal health) and the postnatal environment (housing and neighbourhood conditions) on subsequent health outcomes for the infants born to these women. Data collection will commence during the participants' pregnancy, from approximately 20 weeks gestation. Participants will complete a questionnaire, undergo anthropometric measurements, wear an accelerometer, compile a food diary, and have environmental measures taken within their home. They will also be asked to consent to having a sample of umbilical cord blood taken following delivery of their baby. These data will be complemented by routinely collected electronic data such as health records from GP surgeries, hospital admissions, and child health and development records. Thereafter, participants will be visited annually for follow-up of subsequent exposures and child health outcomes. DISCUSSION: The baseline assessment of EHL will provide information concerning the impact of gestational and postnatal environments on birth outcomes and maternal and infant health. The findings can be used to inform the development of complex interventions targeted at structural, environmental factors, intended to reduce ill-health. Long-term follow-up of the cohort will focus on relationships between environmental exposures and the later development of adverse health outcomes, including obesity and diabetes.

Selected integrative medicine treatments for depression: considerations for women.

This review evaluates the research published between 1966 and 2004 on several integrative treatments for depression, including omega-3 fatty acids, Hypericum perforatum (St. John's Wort), S-adenosyl-methionine, folate, 5-Hydroxytryptophan, acupuncture, exercise, and light therapy, with a particular emphasis on issues pertinent to women. Data from double-blind, placebo-controlled trials support each of these as treatment interventions for depression. We discuss both the strength of the evidence for each treatment and methodological issues related to interpretation of efficacy. Available data pertaining to considerations in women, including use during pregnancy and breastfeeding and interactions with hormonal therapies are discussed. The reviewed treatments deserve further research. Their appropriate place in the armamentarium of depression treatments for women must be defined. An evidence-based integrative medicine approach brings together treatment options with proven efficacy and the public's desire for complementary and alternative medicine treatments.