Pre-translational regulation of luteinizing hormone receptor in follicular somatic cells of cattle

AbstractDifferential regulation of LHR in theca cells (TC) and granulosa cells (GC) is important for normal follicular development. Unlike TC, GC only acquire LH-responsiveness during the later stages of antral follicle development. This study tested the hypothesis that differential LH-responsiveness in these two cell types may be due, in part, to shifts in cellular patterns of alternatively spliced LHR mRNA transcripts which may not be obvious from analysis of total LHR gene expression. It also further explored the role of translation inhibition by an LHR binding protein (LHBP), normally associated with the production of endogenous cholesterol. LHR mRNA variation arises as a result of the alternative splicing of two variable deletion sites (VDS) designated 5′ VDS and 3′ VDS, and it was proposed that differences in cell sensitivity to LH may be due in part to variations in the pattern of the mRNA expression of the receptor variants. The outcomes of the present study support a dynamic multi-facetted regulation of LHR during pre-translation. Not only did the ratio between variants change during antral follicle growth and in vitro cell differentiation but also between TC and GC. Regulation could also be linked to LH concentration feedback mechanisms as the absence of LH caused cultured TC to markedly up-regulate amounts of LHR mRNA. In both TC and GC, LHR mRNA was greatly reduced after treatment to block mevalonate production in the de novo cholesterol pathway, adding further support for a regulatory mechanism linked to enriched cellular amounts of mevalonate kinase

Health knowledge and infection control by event horse owners

Infection control and quarantine measures are essential to minimise the impact of equine infectious diseases, but their uptake by equestrians involved with sport horses in the United Kingdom is undocumented. Using a questionnaire, this study aimed to: i) determine the knowledge and practices of eventing equestrians about their horses’ health and the clinical signs of endemic and exotic equine infectious diseases and ii) assess existing infection control and quarantine measures on event horse yards. A questionnaire was designed, piloted and distributed electronically over six weeks. The target respondents were equestrians involved in eventing, including jumping their horses over fences either 100cms. Data were described qualitatively and analysed statistically to identify any relationships between selected parameters. A total of 146 responses were analysed. The majority of respondents were female, involved as amateurs in eventing and their veterinary surgeon was the first choice for advice. To assess temperature,owners often used touch, but rectal temperature was taken rarely. The majority of horses were vaccinated against tetanus and influenza but a minority against EHV-1/-4. Most respondents identified the clinical signs of influenza and Streptococcus equi, but were less certain about EHV-1/-4. Only 30.1% of respondents had access to quarantine facilities, which were significantly more likely to be available in professional yards (p=0.043) and in riders competing above the 100cm level (p=0.0003). Professionals competing their horses above 100cm were significantly more likely to have quarantine facilities (p<0.05). A majority of yards with quarantine facilities isolated new horses for 2-4 weeks. Facilities included a separate stable (81.8%) or field (68.2%), but separate equipment (47.7%) and access to disinfectant (36.4%) were available less frequently. In conclusion, amateur equestrians involved in eventing require better education on the routine use of infection control and quarantine measures to minimise the impact of equine infectious diseases and thus strengthen infection control nationally

The efficacy of supplying supplemental cobalt, selenium and vitamin B₁₂ via the oral drench route in sheep

Cobalt and selenium are essential trace elements required for ruminants. There are many different methods of supplementation available to sheep including boluses, in feed, free access minerals, pasture dressing and oral drenches. Recent changes in European legislation have resulted in a reduction in the maximum permitted level (MPL) of cobalt to be included in ruminant diets from 2 mg/kg at 88% DM to 1 mg/kg at 88% DM with a suggested supplementary rate of 0.3 mg Co/kg at 88% DM. This study aimed to determine the efficacy of cobalt plus/minus vitamin B₁₂ and selenium supplementation from oral drenching over a 13 day time period. Seven groups of grass silage fed Suffolk cross mule lambs (n=56) were randomly allocated to one of 7 treatments, a 5 ml oral drench containing 700, 2300, 12000 mg Co/l, with or without 2300 mg vitamin B₁₂/l in a factoral design, with an additional control group which received no drench. All drenches also contained selenium at 625 mg/l. Lambs were weighed on days 0 and 13. The lambs were sampled by jugular venepuncture on days 0, 1, 2, 3, 4, 5, 6, 7, 9, 11 and 13, with samples analysed for plasma selenium and cobalt by ICP-MS, erythrocyte glutathione peroxidase by colourimetric assay and vitamin B₁₂ by immunoassay. The results showed that cobalt via a drench was able to raise plasma cobalt in a dose dependent manner, with each dose level significantly higher than the previous level at all post drenching time points (P<0.05). However, the cobalt containing drenches did not significantly raise vitamin B₁₂ concentrations in the plasma. Vitamin B₁₂ containing drenches were able to elevate vitamin B₁₂ in the plasma for a period of 7 days (P<0.05). The selenium content of the drench was able to significantly raise the plasma selenium from day 1 throughout the rest of the trial (day 13) (P<0.001). A significant increase in erythrocyte glutathione peroxidase activity of the sheep did eventually occur at day 13 (P<0.05). In summary, the addition of vitamin B₁₂ via the drench route resulted in a short term elevation (7 days) of plasma vitamin B₁₂ concentrations whereas cobalt sulphate alone was unable to significantly raise vitamin B₁₂ concentrations within this trial in sheep with marginally adequate cobalt status, despite elevating plasma cobalt concentrations. The drench route was also able to elevate plasma selenium concentrations from a marginal selenium status

Health knowledge and infection control by event horse owners

Infection control and quarantine measures are essential to minimise the impact of equine infectious diseases, but their uptake by equestrians involved with sport horses in the United Kingdom is undocumented. Using a questionnaire, this study aimed to: i) determine the knowledge and practices of eventing equestrians about their horses’ health and the clinical signs of endemic and exotic equine infectious diseases and ii) assess existing infection control and quarantine measures on event horse yards. A questionnaire was designed, piloted and distributed electronically over six weeks. The target respondents were equestrians involved in eventing, including jumping their horses over fences either 100cms. Data were described qualitatively and analysed statistically to identify any relationships between selected parameters. A total of 146 responses were analysed. The majority of respondents were female, involved as amateurs in eventing and their veterinary surgeon was the first choice for advice. To assess temperature,owners often used touch, but rectal temperature was taken rarely. The majority of horses were vaccinated against tetanus and influenza but a minority against EHV-1/-4. Most respondents identified the clinical signs of influenza and Streptococcus equi, but were less certain about EHV-1/-4. Only 30.1% of respondents had access to quarantine facilities, which were significantly more likely to be available in professional yards (p=0.043) and in riders competing above the 100cm level (p=0.0003). Professionals competing their horses above 100cm were significantly more likely to have quarantine facilities (p<0.05). A majority of yards with quarantine facilities isolated new horses for 2-4 weeks. Facilities included a separate stable (81.8%) or field (68.2%), but separate equipment (47.7%) and access to disinfectant (36.4%) were available less frequently. In conclusion, amateur equestrians involved in eventing require better education on the routine use of infection control and quarantine measures to minimise the impact of equine infectious diseases and thus strengthen infection control nationally

Mimicking the Impact of Infant Tongue Peristalsis on Behavior of Solid Oral Dosage Forms Administered During Breastfeeding.

An in vitro simulation system was developed to study the effect of an infant's peristaltic tongue motion during breastfeeding on oral rapidly disintegrating tablets in the mouth, for use in rapid product candidate screening. These tablets are being designed for use inside a modified nipple shield worn by a mother during breastfeeding, a proposed novel platform technology to administer drugs and nutrients to breastfeeding infants. In this study, the release of a model compound, sulforhodamine B, from tablet formulations was studied under physiologically relevant forces induced by compression and rotation of a tongue mimic. The release profiles of the sulforhodamine B in flowing deionized water were found to be statistically different using 2-way ANOVA with matching, when tongue mimic rotation was introduced for 2 compression levels representing 2 tongue strengths (p = 0.0013 and p < 0.0001 for the lower and higher compression settings, respectively). Compression level was found to be a significant factor for increasing model compound release at rotational rates representing nonnutritive breastfeeding (p = 0.0162). This novel apparatus is the first to simulate the motion and pressures applied by the tongue and could be used in future infant oral product development.This work was made possible through the generous support of the Saving Lives at Birth partners: the United States Agency for International Development (USAID), the Government of Norway, the Bill & Melinda Gates Foundation (grant number: OPP1129832), Grand Challenges Canada, and the UK Department for International Development (DFID); as well as the Gates Cambridge Trust.This is the final version of the article. It first appeared from Elsevier via http://dx.doi.org/10.1016/j.xphs.2016.08.00

Liver copper concentrations in cull cattle in the UK: are cattle being copper loaded?

With the release of the Department for the Environment, Food and Rural Affairs/Advisory Committee on Animal Feed Guidance Note for Supplementing Copper to Bovines it was noted that the current copper status of the national herd was not known. Liver samples were recovered from 510 cull cattle at a single abattoir across a period of three days. The samples were wet-ashed and liver copper concentrations determined by inductively coupled plasma mass spectrometry analysis. Breed, age and previous location information were obtained from the British Cattle Movement Service. Dairy breeds had higher liver copper concentrations than beef breeds. Holstein-Friesian and ‘other’ dairy breeds had 38.3 per cent and 40 per cent of cattle above the Animal Health and Veterinary Laboratories Agency (AHVLA) reference range (8000 µmol/kg dry matter), respectively, whereas only 16.9 per cent of animals in the combined beef breeds exceeded this value. It was found that underlying topsoil copper concentration was not related to liver copper content and that age of the animal also had little effect on liver concentration. In conclusion, over 50 per cent of the liver samples tested had greater-than-normal concentrations of copper with almost 40 per cent of the female dairy cattle having liver copper concentrations above the AHVLA reference range, indicating that a significant proportion of the UK herd is at risk of chronic copper toxicity

Health knowledge and infection control by event horse owners

Infection control and quarantine measures are essential to minimise the impact of equine infectious diseases, but their uptake by equestrians involved with sport horses in the United Kingdom is undocumented. Using a questionnaire, this study aimed to: i) determine the knowledge and practices of eventing equestrians about their horses’ health and the clinical signs of endemic and exotic equine infectious diseases and ii) assess existing infection control and quarantine measures on event horse yards. A questionnaire was designed, piloted and distributed electronically over six weeks. The target respondents were equestrians involved in eventing, including jumping their horses over fences either 100cms. Data were described qualitatively and analysed statistically to identify any relationships between selected parameters. A total of 146 responses were analysed. The majority of respondents were female, involved as amateurs in eventing and their veterinary surgeon was the first choice for advice. To assess temperature,owners often used touch, but rectal temperature was taken rarely. The majority of horses were vaccinated against tetanus and influenza but a minority against EHV-1/-4. Most respondents identified the clinical signs of influenza and Streptococcus equi, but were less certain about EHV-1/-4. Only 30.1% of respondents had access to quarantine facilities, which were significantly more likely to be available in professional yards (p=0.043) and in riders competing above the 100cm level (p=0.0003). Professionals competing their horses above 100cm were significantly more likely to have quarantine facilities (

Characterising the disintegration properties of tablets in opaque media using texture analysis.

Tablet disintegration characterisation is used in pharmaceutical research, development, and quality control. Standard methods used to characterise tablet disintegration are often dependent on visual observation in measurement of disintegration times. This presents a challenge for disintegration studies of tablets in opaque, physiologically relevant media that could be useful for tablet formulation optimisation. This study has explored an application of texture analysis disintegration testing, a non-visual, quantitative means of determining tablet disintegration end point, by analysing the disintegration behaviour of two tablet formulations in opaque media. In this study, the disintegration behaviour of one tablet formulation manufactured in-house, and Sybedia Flashtab placebo tablets in water, bovine, and human milk were characterised. A novel method is presented to characterise the disintegration process and to quantify the disintegration end points of the tablets in various media using load data generated by a texture analyser probe. The disintegration times in the different media were found to be statistically different (P<0.0001) from one another for both tablet formulations using one-way ANOVA. Using the Tukey post-hoc test, the Sybedia Flashtab placebo tablets were found not to have statistically significant disintegration times from each other in human versus bovine milk (adjusted P value 0.1685).This work was made possible through the generous support of the Saving Lives at Birth partners: the United States Agency for International Development (USAID), the Government of Norway, the Bill & Melinda Gates Foundation, Grand Challenges Canada and the UK Department for International Development (DFID).This is the accepted manuscript. The final version is available at http://www.sciencedirect.com/science/article/pii/S0378517315002392#

The efficacy of supplying supplemental cobalt, selenium and vitamin B₁₂ via the oral drench route in sheep

Cobalt and selenium are essential trace elements required for ruminants. There are many different methods of supplementation available to sheep including boluses, in feed, free access minerals, pasture dressing and oral drenches. Recent changes in European legislation have resulted in a reduction in the maximum permitted level (MPL) of cobalt to be included in ruminant diets from 2 mg/kg at 88% DM to 1 mg/kg at 88% DM with a suggested supplementary rate of 0.3 mg Co/kg at 88% DM.This study aimed to determine the efficacy of cobalt plus/minus vitamin B₁₂ and selenium supplementation from oral drenching over a 13 day time period.Seven groups of grass silage fed Suffolk cross mule lambs (n=56) were randomly allocated to one of 7 treatments, a 5 ml oral drench containing 700, 2300, 12000 mg Co/l, with or without 2300 mg vitamin B₁₂/l in a factoral design, with an additional control group which received no drench. All drenches also contained selenium at 625 mg/l. Lambs were weighed on days 0 and 13. The lambs were sampled by jugular venepuncture on days 0, 1, 2, 3, 4, 5, 6, 7, 9, 11 and 13, with samples analysed for plasma selenium and cobalt by ICP-MS, erythrocyte glutathione peroxidase by colourimetric assay and vitamin B₁₂ by immunoassay.The results showed that cobalt via a drench was able to raise plasma cobalt in a dose dependent manner, with each dose level significantly higher than the previous level at all post drenching time points (