Downer cows: a reanalysis of an old data set.

AIMS: To compare the performance of two predictive models for the survival of downer cows. METHODS: The first model had been developed in 1987 using a dataset containing missing values, while the second, new model was developed on the same dataset but using modern data imputation and analytical methods. Missing data were imputed using multiple imputation by chained equations and a logistic regression model fitted to the imputed data, with survival or not as the outcome variable. The predictive ability of the model built on the imputed data was contrasted with the original prognostic model by testing them both on a second smaller but complete data set, collected contemporaneously with the development of the original model but from a different region of New Zealand. Sensitivity, specificity, accuracy, and cut point for the two models were calculated. RESULTS: The original 1987 model had a slightly higher accuracy than that of the new one with a sensitivity of 0.85 (95% CI = 0.72-0.94) and a specificity of 0.82 (95% CI = 0.7-0.91), using a cut point for the probability of survival = 0.313. CONCLUSIONS: The original prognostic formula published by Clark et al. in 1987 performed as well as a modern model built on an imputed data set. CLINICAL RELEVANCE: The use of a prognostic test based on the Clark model should remain an important part of the clinical examination of downer cows by New Zealand veterinarians.Abbreviations: AUC: Area under the curve; AST: Aspartate transaminase activity; CK: Creatine phosphokinase activity; GAM: Generalised additive model; NSAID: Non-steroidal-anti-inflammatory drugs; PCV: Packed cell volume.

Exploring the perceptions and acceptability of an integrated lifestyle database for public health research and service commissioning: a qualitative study.

AIM: Public health lifestyle databases at local authority level are currently poorly aligned across the UK. The integration of lifestyle databases at a regional level could provide a rich resource to support research and help inform public health leads and service commissioners in improving service delivery, facilitating decision-making and developing key public health policies. Prior to its implementation, the acceptability of an integrated lifestyle database should be explored. The aim of this study was to consult with public health stakeholders to explore the acceptability of developing and implementing a regional integrated lifestyle database across four key areas of public health: smoking cessation, diet, physical activity and alcohol consumption. METHOD: Qualitative interviews were conducted with public health stakeholders recruited from across the East Midlands region of England. All interviews were conducted using video conferencing software and recorded, transcribed, and analysed using the Framework approach. Sixteen public health stakeholders were purposively identified and invited to participate in interviews. RESULTS: Stakeholders viewed the integrated database as having potential to support research, service development and commissioning decisions. Barriers such as providers' reluctance to reveal their business strategies to rival organisations, cost of setting up and running the proposed database, complex information-sharing and governance were identified. CONCLUSION: An integrated lifestyle database has the potential to support research and service commissioning regionally. However, several barriers were identified that must be addressed prior to the development and implementation of an integrated database.

The first week following insemination is the period of major pregnancy failure in pasture-grazed dairy cows.

A 60% pregnancy success for inseminations is targeted to optimize production efficiency for dairy cows within a seasonal, pasture-grazed system. Routine measures of pregnancy success are widely available but are limited, in practice, to a gestation stage beyond the first 28 d. Although some historical data exist on embryonic mortality before this stage, productivity of dairy systems and genetics of the cows have advanced significantly in recent decades. Accordingly, the aim was to construct an updated estimate of pregnancy success at key developmental stages during the first 70 d after insemination. Blood samples were collected for progesterone concentrations on d 0 and 7. A temporal series of 4 groups spanning fertilization through d 70 were conducted on 4 seasonal, pasture-grazed dairy farms (n = 1,467 cows) during the first 21 d of the seasonal breeding period. Morphological examination was undertaken on embryos collected on d 7 (group E7) and 15 (group E15), and pregnancy was diagnosed via ultrasonography on approximately d 28 and 35 (group E35) as well as d 70 (group E70). Fertilization, embryo, and fetal evaluation for viability established a pregnancy success pattern. Additionally, cow and on-farm risk factor variables associated with pregnancy success were evaluated. We estimated pregnancy success rates of 70.9%, 59.1%, 63.8%, 62.3%, and 56.7% at d 7, 15, 28, 35, and 70, respectively. Fertilization failure (15.8%) and embryonic arrest before the morula stage (10.3%) were the major developmental events contributing to first-week pregnancy failures. Embryo elongation failure of 7% contributed to pregnancy failure during the second week. The risk factors for pregnancy success that were related to the cows included interval between calving and insemination, and d-7 plasma progesterone concentrations, whereas insemination sire was associated with pregnancy outcome. Most pregnancy failure occurs during the first week among seasonal-calving pasture-grazed dairy cows.

Immediate reinforcement increased duration of time riding the stationary bicycle in children with autism spectrum disorder: a pilot study.

The necessity of promoting physical activity in individuals with Autism Spectrum Disorder (ASD) has been emphasized for decades. One of the barriers to participate in physical activity for individuals with ASD is limited interest and motivation. Therefore, understanding the motivation to exercise in this population is important. The objective was to determine the effect of using contingent reinforcement in the form of watching a preferred DVD to increase duration of time pedalling on a stationary bicycle within their predetermined target heart rate zone (THRZ) in children with ASD. Using a crossover design, seven participants (11 2.7 years) who were diagnosed with ASD were randomly assigned to either Group A or B. Time spent pedalling on a bicycle within the THRZ was analysed using a linear mixed-effect model with Bonferroni adjustments. The results showed that the DVD intervention motivated children with ASD to exercise for more than 10 minutes in moderate to vigorous physical activity compared to when they were exercising without watching a DVD. This result is significant as number of studies have revealed that 10 minutes of exercise could bring improvements in activities of daily living such as behaviors and academic performance in school.

Where next for the design, delivery, and evaluation of community-based physical activity prescription? Emerging lessons from the United Kingdom.

Despite widespread use, community-based physical activity prescription is controversial. Data limitations have resulted in a lack of clarity about what works, under what circumstances, and for whom, reflected in conservative policy recommendations. In this commentary we challenge a predominantly negative discourse, using contemporary research to highlight promising findings and "lessons learnt" for design, delivery, and evaluation. In doing so, we argue for the importance of a more nuanced approach to future commissioning and evaluation. Novelty: Amalgamating learning from multiple research teams to create recommendations for advancing physical activity prescription.

Rabies risk and use of post-exposure prophylaxis associated with dog bites in Tennessee.

The canine variant of the rabies virus has been eliminated in the United States. Among the public and many healthcare providers, however, dog bites are still associated with risk for rabies transmission. This study examined the risk of rabies in biting dogs and the use of rabies post-exposure prophylaxis (rPEP) for dog bite victims in Tennessee. The study included a retrospective analysis of laboratory testing requisitions for dogs from 2002 to 2016, collection of clinical data on confirmed rabies-positive dogs from 2008 to 2016 and analysis of hospital discharge data for rPEP from 2007 to 2014. Among dogs submitted for rabies testing, those having a recent history of biting were significantly less likely to test positive for rabies than dogs with no reported bite (OR = 0.01; 95% CI [0.003-0.04]). The most common clinical signs reported among rabies-positive dogs were anorexia, dysphagia, ataxia, limb paresis or paralysis, and lethargy; aggressiveness was uncommon. Among hospital patients with an animal-related injury who received rPEP, more than half (52%) presented with dog bites. These data show that laboratory submissions for rabies testing and prescriptions for rPEP do not reflect the epidemiology of rabies in Tennessee. Education and outreach targeting the public and healthcare providers should emphasize the animal species and situations associated with a greater risk for rabies transmission, such as bites from rabies reservoir species or animals exhibiting signs of neurologic disease.

Temporal and spatial heterogeneity in milk and immune-related gene expression during mammary gland involution in dairy cows.

The aim of this study was to investigate heterogeneity in tissue morphology, milk protein and immune-related gene expression, and apoptosis of epithelial cells in the lactating and involuting mammary glands of the dairy cow. Mammary tissue from different regions of the gland (alveolar, cisternal, and peripheral) was collected postmortem from nonpregnant, pasture-fed, Holstein-Friesian primiparous cows in mid-lactation that were killed at different time points postmilking: 0, 6, 12, 18, 24, 36, and 72 h (n = 6 per time point). The CSN1NS1 and LALBA mRNA was decreased in alveolar, cisternal, and peripheral tissue by 12 to 36 h postmilking. In contrast, lactoferrin (LF) and mammary serum amyloid A3 (M-SAA3) mRNA was increased in these regions by 36 to 72 h. During lactation, more variability was present in gene expression in alveolar tissue between cows and between quarters within a cow, than within quarters. Histological analysis indicated the alveolar tissue from lactating cows was mostly uniform in structure; however, in situ hybridization indicated that although most of the alveolar tissue expressed milk proteins, the level of expression varied within and between alveoli. This heterogeneity became more pronounced with involution and with increasing regions of alveoli expressing lactoferrin, indicating that alveoli enter involution asynchronously. The peripheral and cisternal tissue had more variability in gene expression between cows compared with the alveolar tissue. The M-SAA3 signal was more intense in the cisternal tissue and less intense than the peripheral compartment compared with LF particularly in the earlier time points. In addition, between cows within the later time points, differences were observed in tissue morphology, the levels of milk protein and immune-related gene expression, and phosphorylated signal transducer and activator of transcription (STAT) 5-P and STAT3-P proteins, and degree of apoptosis, indicating that involution of the mammary gland occurs at different rates between cows. Understanding the mechanisms initiating the process of involution of the mammary gland provides an opportunity for enhancing milk production of the dairy cow.

Blood borne virus testing of 2250 patients in an unusual, repeated dental patient notification exercise: challenges faced and lessons learnt.

OBJECTIVES: The objectives of this study were to ascertain the risk created for patients of two dental practices where infection control was found to be inadequate, and if the risk was deemed to be significant, initiate an investigation involving notification and blood borne virus (BBV) testing to establish if any patient-to-patient BBV transmissions had occurred as a result of these infection control breaches. STUDY DESIGN: A case study. METHODS: A public health investigation and patient notification. Investigations involved practice inspections, staff interviews and examination of invoices. The practices were not fully cooperative during the investigation and provided misleading information regarding the allegations. This led to two patient notification exercises, as more serious breaches were uncovered following the first notification exercise. Risk assessments of BBV transmission likelihood were undertaken and informed the nature of the advice given to patients. RESULTS: The health board wrote to 5100 patients informing them of the situation. BBV testing was offered in the second notification exercise and 2250 patients opted to be tested for HIV, hepatitis B and hepatitis C. There were no new cases of HIV or hepatitis B but less than five patients were found to be positive for hepatitis C. None of these cases were proven to have contracted their infection as a result of the dental infection control lapses. CONCLUSIONS: This incident was unusual in that the practice was found to be repeatedly and knowingly putting patients at risk, and attempts were made to cover up breaches during the investigation. In future, health boards would benefit from a risk assessment tool to aid decision making regarding notification exercises, and whether testing is indicated where risk to patients is low. This would help ensure that notification exercises do more good than harm.

Animal physiology and genetic aspects of ryegrass staggers in grazing sheep.

Ryegrass staggers (RGS) is a metabolic disease of herbivores, caused by the ingestion of perennial ryegrass (Lolium perenne L.) containing a fungal endophyte (Neotyphodium lolii) which produces a tremorgenic toxin, lolitrem B. RGS has a major economic impact for agriculture in New Zealand as well as internationally. Management of RGS in grazing sheep can be problematic, and there is an incomplete knowledge of the interaction between the toxin and the grazing animal. This review is focused on recent advances in understanding the molecular physiology of RGS in the affected animal as well as the influence of animal genetics on the degree of susceptibility to RGS. Investigations to date suggest that the primary target for toxin is the large conductance, calcium-activated, potassium (BK) channel, resulting in disruption of neuromuscular junction signalling. Genetic investigation has established the existence of genes influencing resistance to RGS, however their identity has not been confirmed and their impact has not been established. Studies to date suggest that a multi-gene selection approach will be necessary in order to develop an effective selection tool for use in the agricultural industries.

Cell survival signaling in the bovine mammary gland during the transition from lactation to involution.

In dairy cows, mammary gland involution, and thus a decline in milk production, occurs following peak lactation. To examine the cell signaling pathways regulating involution of the mammary gland, signal transducer and activator of transcription factors (STAT5 and 3), suppressors of cytokine signaling (SOCS1-3 and CIS), insulin-like growth factors (IGF1 and 2), and protein kinase B (Akt) were examined. Mammary involution was induced by termination of milking, and alveolar tissue was collected from 52 nonpregnant, primiparous, mid-lactation Holstein-Friesian cows killed at 0, 6, 12, 18, 24, 36, 72, and 192h postmilking. Qualitative immunohistochemistry showed that activated (phosphorylated) STAT5-P was localized in nuclei of mammary epithelial cells at the early time points, with detection levels decreasing by 24h postmilking. In contrast, STAT3-P was barely detectable at the early time points, with detection levels increasing following longer postmilking periods. This was supported by Western analysis, which showed a decline in STAT5 and STAT5-P protein levels by 24h postmilking, no change in STAT3 levels, and an increase in STAT3-P protein (barely detectable at the early time points) by 72h postmilking. Quantitative real-time reverse transcription PCR analysis showed SOCS1 and SOCS3 mRNA increased by 72h postmilking compared with 6h postmilking. The SOCS2 mRNA remained unchanged across the time series, whereas CIS decreased by 18h postmilking and remained lower compared with that at 6h postmilking until 72h postmilking. The IGF1 mRNA increased by 192h postmilking, whereas IGF2 mRNA decreased by 18h postmilking compared with 6h postmilking. The IGFBP5 mRNA and protein levels of Akt and Akt-P remained unchanged over the time series. These results show that reciprocal activation of STAT5 and STAT3 occurs at the onset of mammary gland involution in the bovine, albeit at a slower rate than in rodents. Mathematical modeling of the pathways indicated that activated STAT3 could block the STAT5 pathway by upregulating SOCS3. The regulation of IGF1-Akt signaling suggests that by 192h postmilking in dairy cows, the involution process is still in the reversible phase, with quiescent mammary epithelial cells not yet in the senescent phase.

Ultrasonography to investigate the effect of supplementing whole milk with complex carbohydrates and specific amino acids on curd retention in the abomasum of dairy calves.

AIM: To determine whether the retention time of curd in the abomasum of calves was influenced by supplementing milk with a plant-derived carbohydrate and amino acid supplement, evaluated non-invasively using ultrasonography. METHODS: Female dairy calves aged between 2-6 days of age were sourced from a commercial farm in March 2013. All calves were fed whole milk until weaning (4 L per day); 21 calves were supplemented with a probiotic until 18 days of age, and thereafter with a plant-derived complex carbohydrate and amino acid supplement until weaning, and 22 calves were just fed whole milk. Treatment groups were balanced for age, weight and breed. At 9-14, 24-29 and 52-57 days of age, the abomasum of each calf was examined using ultrasonography immediately before and after feeding, 1 and 2 hours after feeding, and then at 30 minute intervals until curd was no longer visible in the abomasum. Abomasal volume and curd size were recorded to assess retention time of curd in the abomasum. RESULTS: At 9-14 days of age, mean retention time of curd in the abomasum was similar (4.6 hours) in both groups. At 24-29 days of age, when the supplemented calves had been receiving the supplement for approximately 10 days, mean curd retention time was longer by 1.4 (SE 0.28) hours in supplemented compared with unsupplemented calves (p<0.001). At 52-57 days of age, mean retention time was longer by 0.7 (SE 0.34) hours compared to unsupplemented calves (p=0.05). CONCLUSION: Using ultrasonography, changes in abomasal content could be followed non-invasively over time and it was demonstrated that the plant-derived complex carbohydrate supplement increased the curd retention time in the abomasum. We speculate that the increased retention time enables an increased availability of nutrients following a more complete digestion of milk, thereby improving animal performance.

Formulations for delivering Trichoderma atroviridae spores as seed coatings, effects of temperature and relative humidity on storage stability.

AIMS: We aimed to evaluate different formulations for their ability to adhere Trichoderma atroviridae spores to wheat seeds, and promote survival during storage at a range of temperatures and relative humidities (RH). METHODS AND RESULTS: We tested a range of formulations for their ability to adhere T. atroviridae spores to wheat seeds. Treated seeds were stored for 6 months at a range of temperatures and RH, and spore viability among formulation was compared over time. Spore survival within formulations interacted significantly with environmental conditions. Notably, under optimum conditions (low temperatures and RH) best spore survival was recorded with a xanthan-gum-based formulation. Conversely under suboptimum conditions (high temperatures and RH), survival of spores was best in a waxy-starch formulation, but very poor in the xanthan-gum formulation. CONCLUSIONS: These results indicate that T. atroviridae spores can be effectively delivered on to seeds and that a xanthan-gum formulation is promising when optimal storage conditions can be maintained. SIGNIFICANCE AND IMPACT OF THE STUDY: Most published formulation papers/patents only report survival of organisms over time at a single or limited number of temperatures and RH. For the first time, this study shows how different formulations are better suited to certain temperature and RH combinations.

The effect of milking reinitiation following extended nonmilking periods on lactation in primiparous dairy cows.

In dairy cows, extended periods of nonmilking results in reduced milk secretion, modifications in milk composition, and eventually involution of the mammary glands. The aim of this study was to determine the effect of extended nonmilking periods on the recovery of milk yield and composition, and levels of prolactin and insulin-like growth factor-I in pasture-fed cows after resuming milking. Pasture-fed, primiparous, nonpregnant, Friesian dairy cows at mid lactation (mean ± standard deviation, 97 ± 2d in milk, 14.0 ± 2.5 L/d) were divided into 3 groups (n=6 per group). The cows were subjected to nonmilking periods of 7, 14, or 28d. Twice-daily milking was resumed for 7d following the nonmilking periods. Milk yield recoveries at the end of the 7-d remilking period were 91, 51, and 29% for the 7, 14, and 28-d nonmilked groups, respectively. The somatic cell count declined to less than 400,000 cells/mL by d 3 and 6 of remilking for the 7- and 14-d-nonmilked groups, respectively, but remained greater than 800,000 cells/mL in the 28-d-nonmilked group through the 7-d remilking period. By d 7 of remilking, the somatic cell count for the 7-d-nonmilked group was not different from pretrial values. Upon remilking, the milk fat content returned to pretrial values for the 7- and 14-d-nonmilked groups, although it remained lower than pretrial for the 28-d-nonmilked group. All 3 nonmilked groups had a higher milk protein content following 7d of remilking, compared with pretrial values. The lactose content returned to pretrial values for the 7-d-nonmilked group but remained lower for the 14- and 28-d-nonmilked groups. Circulating prolactin concentrations increased once remilking was resumed, compared with the pretrial and nonmilking periods. Prolactin concentrations did not majorly differ between the groups, with the levels upon 7d of remilking remaining higher than the pretrial concentrations and the nonmilked periods. Plasma concentrations of insulin-like growth factor-I increased during the nonmilking period and were greater in all 3 nonmilked groups on d 1 of remilking than pretrial values and returned to pretrial concentrations following remilking for the 7-d-nonmilked group, whereas the 14- and 28-d-nonmilked groups remained higher than the pretrial values. These data indicate that the process of involution is fully reversed after remilking following 7d of milk stasis but more extended periods of nonmilking prevent the complete recovery of lactation. However, even after 28d of milk stasis, the milk synthesis capacity of the mammary gland could still be partially recovered.

Genome-wide DNA methylation analysis: no evidence for stable hemimethylation in the sheep muscle genome.

The importance of maintaining DNA methylation patterns and faithful transmission of these patterns during cell division to ensure appropriate gene expression has been known for many decades now. It has largely been assumed that the symmetrical nature of CpG motifs, the most common site for DNA methylation in mammals, together with the presence of maintenance methylases able to methylate newly synthesised DNA, ensures that there is concordance of methylation on both strands. However, although this assumption is compelling in theory, little experimental evidence exists that either supports or refutes this assumption. Here, we have undertaken a genome-wide single-nucleotide resolution analysis to determine the frequency with which hemimethylated CpG sites exist in sheep muscle tissue. Analysis of multiple independent samples provides strong evidence that stably maintained hemimethylation is a very rare occurrence, at least in this tissue. Given the rarity of stably maintained hemimethylation, next-generation sequencing data from both DNA strands may be carefully combined to increase the accuracy with which DNA methylation can be measured at single-nucleotide resolution.

Influence of pathogenic bacteria species present in the postpartum bovine uterus on proteome profiles.

In the first 2-3 weeks after parturition >90% of dairy cows will have some form of uterine infection. Uterine contamination with pathogens, such as Trueperella (formerly Arcanobacterium) pyogenes increases the risk of developing more severe endometritis, which can reduce conception rates. In this study, we compared the uterine proteome of cows infected with Trueperella pyogenes with that of uninfected cows, using 2D gel electrophoresis, and identified annexins A1 and A2 (ANXA1 and ANXA2), apolipoprotein A-1, calprotectin (S100A9), cathelicidin, enolase 1 (ENO1), peptidoglycan recognition protein 1 (PGLYRP1), phosphoglycerate mutase 1 (PGAM1), serine dehydratase (SDS) and serine protease inhibitors (SERPIN) B1, B3 and B4 proteins as differing in abundance in endometritis. Subsequently, levels of ten of these proteins were monitored in uterine samples collected from a herd of lactating, dairy cows at 15 and 42 days post-partum (DPP). The levels were compared with the cytology scores of the samples and the bacterial species isolated from the uterus. Cathelicidin, PGLYRP1, SERPINB1 and S100A9 levels at 15DPP showed strong positive correlations (r=0.78, 0.80, 0.79, and 0.68 respectively; P<0.001) with % of polymorphonuclear neutrophils (PMN). When compared with other bacterial pathogens identified, Streptococcus agalactiae and Truperella pyogenes induced increased expression of the indicator proteins, suggesting that these organisms may adversely affect the subsequent ability of the cow to conceive. Interestingly, there was no difference in the proportion of cows pregnant at 6 and 17 weeks after start of mating between the cows with high or low %PMN.

The phenotype of an IVF child is associated with peri-conception measures of follicular characteristics and embryo quality.

STUDY QUESTION: Are childhood measures of phenotype associated with peri-conception parental, IVF treatment and/or embryonic characteristics of IVF children? SUMMARY ANSWER: Birthweight, childhood body mass index (BMI) and height of pre-pubertal IVF children were strongly associated with peri-conception factors, including follicular and embryonic characteristics. WHAT IS KNOWN ALREADY: A growing number of studies have identified a range of phenotypic differences between IVF and naturally conceived pre-pubertal children; for example, birthweights are lower following a fresh compared with a thawed embryo transfer. STUDY DESIGN, SIZE, DURATION: This retrospective cohort study included IVF children (n = 96) born at term (>37 weeks) after a singleton pregnancy from the transfer of either fresh or thawed embryos in New Zealand. Between March 2004 and November 2008, these children were subjected to clinical assessment before puberty. PARTICIPANTS/MATERIALS, SETTING, METHODS: Clinical assessment provided anthropometric measures of children aged 3.5-11 years old. Peri-conception factors (n = 36) derived retrospectively from parental, treatment, laboratory and embryonic variables (n = 69) were analysed using multiple stepwise regression with respect to standard deviation scores (SDSs) of the birthweight, mid-parental corrected BMI and height of the IVF children. Data from children conceived from fresh (n = 60) or thawed (n = 36) embryos, that met inclusion criteria and had high-quality data with >90% completeness, were analysed. MAIN RESULTS AND THE ROLE OF CHANCE: Embryo treatment at transfer was identified as a predictor of birthweight with thawed embryos resulting in heavier birthweights than fresh embryos [P = 0.02, 95% confidence interval (CI) fresh minus thawed: -1.047 to -0.006]. Birthweight SDS was positively associated with mid-parental corrected BMI SDS (P = 0.003, slope 0.339 ± 0.100). Four factors were related (P < 0.05) to mid-parental corrected height SDS. In particular, child height was inversely associated with the diameter of lead follicles at oocyte retrieval (P < 0.0001, slope -0.144 ± 0.040) and with the quality score of embryos at transfer (P = 0.0008, slope -0.425 ± 0.157), and directly associated with the number of follicles retrieved (P = 0.05, slope 1.011 ± 0.497). Child height was also positively associated with the transfer of a fresh as opposed to thawed embryo (P < 0.001, 95% CI 0.275-0.750). LIMITATIONS, REASONS FOR CAUTION: More than one embryo was transferred in most cycles so mean development and quality data were used. The large number of variables measured was on a relatively small sample size. Large cohorts from multiple clinics using a variety of treatment protocols and embryology methods are needed to confirm the associations identified and ultimately to test these factors as possible predictors of phenotype. WIDER IMPLICATIONS OF THE FINDINGS: This is the first study to directly associate peri-conception measures of IVF treatment with a pre-pubertal child's phenotype. Demonstration that peri-conception measures relate to a pre-pubertal child's phenotype extends the range of factors that may influence growth and development. These findings, if corroborated by larger studies, would provide invaluable information for practitioners, who may want to consider the impact of ovarian stimulation protocols as well as the quality of the embryo transferred on a child's growth and development, in addition to their impact on pregnancy rate. STUDY FUNDING/COMPETING INTERESTS: This work was supported by grants from the National Research Centre of Growth and Development New Zealand (grant 3682065) and the Australasian Paediatric Endocrine Group (APEG; grant 3621994), as well as a fellowship from Fertility Associates New Zealand awarded to M.P.G. In terms of competing interest, J.C.P is a shareholder of Fertility Associates. M.P.G. currently holds the position of Merck Serono Lecturer in Reproductive Biology. W.S.C. and P.L.H. have also received grants and non-financial support from Novo Nordisk, as well as personal fees from Pfizer that are unrelated to the current study. The other authors have no conflict of interest to declare.

Acanthamoeba encephalitis: A Case Report and Review of Therapy.

BACKGROUND: Acanthamoeba is a rare cause of encephalitis yet is associated with high mortality. Treatment protocols vary greatly and generally include combination therapy across a wide spectrum of antiinfective classes. CASE DESCRIPTION: A 63-year-old male who underwent renal transplantation presented 6 months after transplantation with depressed level of consciousness. Imaging of the head with computerized tomography showed an enhancing lesion suspicious for brain abscess. Biopsy of the lesion showed Acanthamoeba cysts. The patient was treated with sulfadiazine, fluconazole, flucytosine, azithromycin, and miltefosine but without success. We review recently published cases of Acanthamoeba encephalitis with an emphasis on treatment protocols and outcomes. CONCLUSION: Free-living protozoans such as Acanthamoeba are ubiquitous in the environment and should be suspected in immunosuppressed persons who present with central nervous system findings and brain abscess. Biopsy is critical to establish the etiology so that appropriate combination therapy can be deployed.

Implementing a community-based obesity prevention programme: experiences of stakeholders in the north east of England.

Recent literature indicates the potential of community-based obesity prevention programmes in the endeavour to reduce the prevalence of obesity in developed nations. Considerable suggestion and advocacy come from theoretical standpoints and little is known on actual practical application of this type of multi-component health promotion programme. This article explores the experiences of 'implementation' by stakeholders of a large community-based obesity prevention programme, facilitated by a National Health Service Care Trust in the north-east of England, UK. Three stakeholder groups (senior health officials, public health workers and community members) who had administrated and experienced the programme since its conception in 2006 provide perspectives on the aspects of local delivery and receipt. Semi-structured interviews and focus groups were conducted with stakeholders (28 participants in total). The participants felt there were three broad aspects which shaped and constrained the delivery and receipt of the programme, namely partnership working, integration of services and quality issues. Data indicated that it had taken time to establish working partnerships between the multi-agencies involved in the community-based obesity programme. Strategic management would aid the processes of communication and collaboration between agencies and also the local community involved in the administration, delivery and participation of interventions in the programme. Secondly, the way in which the programme is justified and sustained will have to be reviewed, with the intention of using a suitable evaluative framework or tool for monitoring purposes.

Distinct mechanisms of relaxation to bioactive components from chamomile species in porcine isolated blood vessels.

German chamomile (Matricaria recutita L.), a widely-used herbal medicine, has been reported to have a wide range of biological effects, including smooth muscle relaxation. The aim of this study was to compare the effects of representative compounds from chamomile (apigenin, luteolin, (-)-α-bisabolol, farnesene, umbelliferone; 3-30 µM) on vascular tone using porcine coronary and splenic arteries mounted for isometric tension recording in isolated tissue baths and precontracted with the thromboxane-mimetic U46619. Apigenin, luteolin, and (-)-α-bisabolol produced slow, concentration-dependent relaxations in both the coronary and splenic arteries that were not blocked by inhibition of nitric oxide synthase or potassium channels. Removal of extracellular calcium inhibited the relaxations to all three compounds, and these compounds also inhibited calcium re-addition-evoked contractions, indicating that the relaxation response may be mediated through inhibition of calcium influx. Apigenin and luteolin, but not (-)-α-bisabolol, enhanced the relaxation to the nitric oxide donor sodium nitroprusside, indicating that apigenin and luteolin may act to regulate cyclic GMP levels. Umbelliferone produced a rapid, transient relaxation in the splenic artery, but not the coronary artery, that was inhibited by L-NAME and removal of the endothelium, suggesting an influence on nitric oxide production. Farnesene, at concentrations up to 30 µM, was without effect in either blood vessel. In conclusion, hydroxylated compounds (apigenin, luteolin and (-)-α-bisabolol) found in chamomile all caused a slow relaxation of isolated blood vessels through an effect on calcium influx. Umbelliferone, on the other hand, produced a rapid, transient relaxation dependent upon release of nitric oxide from the endothelium.

A qualitative exploration of stakeholder perspectives on a school-based multi-component health promotion nutrition programme.

BACKGROUND: Food for Fitness is an on-going multi-component health promotion programme, delivered in primary and secondary schools by community nutrition assistants. The programme uses nutritional interventions aimed at promoting healthier eating practices for children. This service evaluation investigated the receipt and delivery of the programme, as perceived by local stakeholders who had experienced and administered the service. METHODS: Semi-structured interviews and focus groups were carried out with three key stakeholder groups: health professionals (n = 9), school teachers (n = 10) and senior health officials (n = 3). Qualitative data were transcribed verbatim and received thematic analysis with deductive and inductive processes. RESULTS: Stakeholders reported that the programme contributed to the development of food education and healthy-eating practices of children in the local area. Stakeholders considered that the main concern was the limited capacity and size of the service. They described problems with long-term sustainability in supporting schools with maintaining nutritional interventions, highlighting issues regarding contact, planning and organisation of several interventions. CONCLUSIONS: The findings of the service evaluation inform service management, organisation and ground-level delivery. The use of stakeholder opinion provided contextualised information on the factors that impact on the implementation of the programme. The richness of the qualitative results can guide future planning and provision for similar health promotion nutrition programmes delivered in the school environment.