Alterations of Circulating Biomarkers During Late Term Pregnancy Complications in the Horse Part I: Cytokines.

Equine abortions are attributed to both infectious and noninfectious causes. Clinical extrapolations are often made from the experimental model for ascending placentitis towards other causes of fetal compromise, including various markers of inflammation, including the cytokines IL-2, 5, IL-6, IL-10, IFNγ, and TNF. It is unknown if these cytokine changes are noted under field conditions, or if they increase preceding other pregnancy related complications. To assess this, Thoroughbred mares (n = 702) had weekly blood obtained beginning in December 2013 and continuing until parturition. Fetal membranes were submitted to the UKVDL for complete gross and pathologic assessment and classified as either ascending placentitis (n = 6), focal mucoid placentitis (n = 6), idiopathic abortion (n = 6) or control (n = 20). Weekly serum samples were analyzed via immunoassay for concentrations of IL-2, IL-5, IL-6, IL-10, IFNγ, and TNF. For both focal mucoid placentitis and ascending placentitis, an increase (P < .05) in the concentrations of IL-2, IL-5, IL-6, IL-10, IFNγ, and TNF was noted preceding parturition in comparison to controls. Cytokine profiles preceding idiopathic abortion did not differ from controls. In conclusion, serum cytokines may be considered potential biomarkers for the prediction of placental infection, while no changes in cytokine profiles were noted when noninfectious causes of abortion occurred. Additionally, this is the first study to report an increase in cytokines during the disease process of focal mucoid placentitis, the etiology of which includes Nocardioform placentitis.

Postmortem examination (2016-2017) of weanling and older horses for the presence of select species of endoparasites: Gasterophilus spp., Anoplocephala spp. and Strongylus spp. in specific anatomical sites.

Parasite infections are more quantifiable postmortem than antemortem in horses. Thus a study was carried out examining dead horses for specific parasite species. Most of the weanling and older horses submitted to the University of Kentucky Veterinary Diagnostic Laboratory (UKVDL) for postmortem examination between November 22, 2016 and March 23, 2017 were examined for certain species of internal parasites. The stomach and duodenum from 69 horses were examined for bots (Gasterophilus spp.). Combined data for both Thoroughbred and non-Thoroughbred (16 other than Thoroughbred breeds/mixed breeds) horses revealed that the prevalence of Gasterophilus intestinalis was 19% (n=12) with 2nd instars (x̄ 8.5) and 39% (n=27) with 3rd instars (x̄ 90). The prevalence of Gasterophilus nasalis was 1.5% (n=1) for 2nd instars (x̄ 1) and 7% (n=5) for 3rd instars (x̄ 25). A few third instar G. intestinalis placed in 10% formalin showed slight movement at over two hundred hours later. The cecum and about 25cm of the terminal part of the ileum were examined from 139 horses for tapeworms (Anoplocephala spp.) and large strongyles (Strongylus spp.). The prevalence of A. perfoliata was 44% (n=62) and the average number of specimens per infected horse was 92.5. Strongylus vulgaris and Strongylus edentatus were not found in the gut of any horse.

Food for thought: pilot randomized controlled trial of lay health trainers supporting dietary change to reduce cardiovascular disease in deprived communities.

BACKGROUND: Cardiovascular disease (CVD) accounts for 30% of UK deaths. It is associated with modifiable lifestyle factors, including insufficient consumption of fruit and vegetables (F&V). Lay health trainers (LHTs) offer practical support to help people develop healthier behaviour and lifestyles. Our two-group pilot randomized controlled trial (RCT) investigated the effectiveness of LHTs at promoting a heart-healthy lifestyle among adults with at least one risk factor for CVD to inform a full-scale RCT. METHODS: Eligible adults (aged 21-78 years), recruited from five practices serving deprived populations, were randomized to health information leaflets plus LHTs' support for 3 months (n = 76) versus health information leaflets alone (n = 38). RESULTS: We recruited 114 participants, with 60% completing 6 month follow-up. Both groups increased their self-reported F&V consumption and we found no evidence for LHTs' support having significant added impact. Most participants were relatively less deprived, as were the LHTs we were able to recruit and train. CONCLUSIONS: Our pilot demonstrated that an LHT's RCT whilst feasible faces considerable challenges. However, to justify growing investment in LHTs, any behaviour changes and sustained impact on those at greatest need should be demonstrated in an independently evaluated, robust, fully powered RCT.

Lay food and health worker involvement in community nutrition and dietetics in England: definitions from the field.

BACKGROUND: Community-based food initiatives have developed in recent years with the aim of engaging previously 'hard to reach' groups. Lay workers engaged in community nutrition activities are promoted as a cost-effective mechanism for reaching underserved groups. The primary objective of the study was to explore perceptions and definitions of lay helping within the context of National Health Service (NHS) community nutrition and dietetic services to identify existing terms and definitions and propose an overarching term. METHODS: Interpretive qualitative inquiry; semi-structured interviews with lay food and health worker (LFHW) and NHS professionals employed by community-based programmes, serving 'hard-to-reach' neighbourhoods, across England. RESULTS: In total, 29 professionals and 53 LFHWs were interviewed across 15 of the 18 projects identified. Across all the projects, there was a preference for the use of one of two terms, either Community Food Worker or Community Nutrition Assistant, in reference to lay workers. There was no consensus in terms of a unifying term or definition for this new role. CONCLUSIONS: Current variation in the terms and definitions used for this role is problematic and is hindering development and effective utilization of lay helping within the broad remit of community food and health and dietetics. The umbrella term 'Lay Food and Health Worker' is proposed based upon definitions and interpretations from the field.

Lay food and health worker involvement in community nutrition and dietetics in England: roles, responsibilities and relationship with professionals.

BACKGROUND: Community-based food initiatives have developed in recent years with the aim of engaging previously 'hard to reach' groups. Lay workers engaged in community nutrition activities are promoted as a cost-effective mechanism for reaching underserved groups. The main objective of the study was to explore perceptions and definitions of lay food and health worker (LFHW) helping roles within the context of National Health Service (NHS) community nutrition and dietetic services in order to define the conceptual and practical elements of this new role and examine the interface with professional roles. METHODS: Interpretive qualitative inquiry; semi-structured interviews with LFHW and NHS professionals employed by community-based programmes, serving 'hard-to-reach' neighbourhoods, across England. A total sampling framework was used to capture all existing and 'fully operational' lay food initiatives in England at the commencement of fieldwork (January 2002). FINDINGS: In total, 29 professionals and 53 LFHWs were interviewed across 15 of the 18 projects identified. Although all 15 projects shared a universal goal, to promote healthy eating, this was achieved through a limited range of approaches, characterized by a narrow, individualistic focus. Lay roles spanned three broad areas: nutrition education; health promotion; and administration and personal development. Narratives from both professionals and LFHWs indicated that the primary role for LFHWs was to encourage dietary change by translating complex messages into credible and culturally appropriate advice. CONCLUSIONS: This research confirms the emerging discipline involving lay helping within the NHS and community dietetics. The primary role of LFHWs in the 15 projects involved was to support existing NHS services to promote healthy eating amongst 'hard to reach' communities. The activities undertaken by LFHWs are strongly influenced by professionals and the NHS. Inherent to this is a fairly narrow interpretation of health, resulting in a limited range of practice.

Three-dimensional microscopic image reconstruction of prostatic adenocarcinoma.

CONTEXT: Routine microscopy provides only a 2-dimensional view of the complex 3-dimensional structure that makes up human tissue. Three-dimensional microscopic image reconstruction has not been described previously for prostate cancer. OBJECTIVES: To develop a simple method of computerized 3-dimensional image reconstruction and to demonstrate its applicability to the study of prostatic adenocarcinoma. METHODS: Serial sections were cut from archival paraffin-embedded prostate specimens, immunostained using antikeratin CAM5.2, and digitally imaged. Computer image-rendering software was used to produce 3-dimensional image reconstructions of prostate cancer of varying Gleason grades, normal prostate, and prostatic intraepithelial neoplasia. RESULTS: The rendering system proved easy to use and provided good-quality 3-dimensional images of most specimens. Normal prostate glands formed irregular fusiform structures branching off central tubular ducts. Prostatic intraepithelial neoplasia showed external contours similar to those of normal glands, but with a markedly complex internal arrangement of branching lumens. Gleason grade 3 carcinoma was found to consist of a complex array of interconnecting tubules rather than the apparently separate glands seen in 2 dimensions on routine light microscopy. Gleason grade 4 carcinoma demonstrated a characteristic form of glandular fusion that was readily visualized by optically sectioning and rotating the reconstructed images. CONCLUSIONS: Computerized 3-dimensional microscopic imaging holds great promise as an investigational tool. By revealing the structural relationships of the various Gleason grades of prostate cancer, this method could be used to refine diagnostic and grading criteria for this common tumor.

Community involvement at what cost?--local appraisal of a pan-European nutrition promotion programme in low-income neighbourhoods.

In the UK, government has committed itself to improving health and reducing inequalities in health. For the first time, issues such as food poverty will be addressed by tackling the causes of poverty and wider determinants of ill health. The time has never been better, therefore, for health and local authorities to work collaboratively to promote and improve health. Community involvement is also paramount to sustainable programmes. However, such a dramatic shift in policy and greater emphasis on public health requires health professionals themselves to adopt a different approach. The World Health Organization (WHO) recommends a health promotion approach as a framework for action. But despite the existence of this framework there is little evidence that a wider understanding of health promotion and the necessary practical experience has been achieved. This has weakened the potential impact of health promotion and has possibly encouraged inappropriate use of health promotion principles in practice. The European Food and Shopping Research Project (SUPER project) was established under the WHO European network of Healthy Cities to help local projects implement the principles of health promotion (WHO, 1986). This paper describes the SUPER project and its implementation in Liverpool (1989-1997), where levels of unemployment, deprivation and ill health are amongst the highest in the UK. Participation in SUPER is appraised to identify the various benefits and obstacles involved and to identify links with progress at the local level. This appraisal is discussed and the use, and potential misuse, of participatory appraisal techniques to elicit information and mobilize communities is examined.

Tacrolimus: an alternative for graft-versus-host disease prevention.

OBJECTIVE: To evaluate the efficacy of tacrolimus for prevention of graft-versus-host disease (GVHD) in patients receiving allogeneic bone marrow transplants. DATA SOURCES: Published literature was identified through MEDLINE (January 1990-December 1998) using the key words tacrolimus, FK506, graft-versus-host disease, and bone marrow transplant. DATA SYNTHESIS: GVHD associated with allogeneic bone marrow transplant is a serious life-threatening complication. An evaluation of studies using tacrolimus for prevention of GVHD was conducted. CONCLUSIONS: Tacrolimus is effective for the prevention of GVHD in allogeneic bone marrow transplant. Further studies need to be conducted to optimize the dosage schedule and to determine therapeutic ranges, efficacy, and safety.

A quantitative technique for reporting surface degradation patterns of UHMWPE components of retrieved total knee replacements.

A quantitative method of reporting surface degradation of the ultra-high molecular weight polyethylene (UHMWPE) tibial component from retrieved total knee replacements (TKR) was developed. Specific features include a qualitative assessment expressing the patterns in which the damage was detected as well as a quantitative summary of the observed degradation mechanisms. In addition, a method of measuring lower limb alignment changes with time is described and related to the observed damage patterns. Two case studies are presented. One case illustrated that changes in alignment resulted from factors other than wear. The damage observed on the tibial plateau appeared to occur subsequent to the changes in alignment. The second case illustrated that the wear of the UHMWPE tibial insert lead to the changes in the overall lower limb alignment. The methods described provide additional information regarding TKR failure mechanisms compared to reporting methods currently available. In particular, the collection of temporal alignment data at clinical follow-up visits enhanced the assessment of the retrieved TKR.

Manipulation of dopamine receptors alters hypoxic pulmonary vasoconstriction in isolated perfused rat lungs.

Using an isolated, perfused rat lung model, we examined the hypoxic pulmonary vasoconstriction (HPV). We studied the alterations in HPV induced by the selective DA1 receptor agonist, fenoldopam, the selective DA1 antagonist, SCH 23390, as well as a combination of these agents. Fenoldopam significantly attenuated HPV. SCH 23390 had no effect on HPV, but was ableto block the effect of fenoldopam. These data confirm the presence of vasodilatory DA1 receptors in the pulmonary vascular bed. The data further suggest that ongoing DA1 activity may be important in counterbalancing some pathologic pulmonary hypertensive states.

X-ray crystal structure of the protease inhibitor domain of Alzheimer's amyloid beta-protein precursor.

Alzheimer's amyloid beta-protein precursor contains a Kunitz protease inhibitor domain (APPI) potentially involved in proteolytic events leading to cerebral amyloid deposition. To facilitate the identification of the physiological target of the inhibitor, the crystal structure of APPI has been determined and refined to 1.5-A resolution. Sequences in the inhibitor-protease interface of the correct protease target will reflect the molecular details of the APPI structure. While the overall tertiary fold of APPI is very similar to that of the Kunitz inhibitor BPTI, a significant rearrangement occurs in the backbone conformation of one of the two protease binding loops. A number of Kunitz inhibitors have similar loop sequences, indicating the structural alteration is conserved and potentially an important determinant of inhibitor specificity. In a separate region of the protease binding loops, APPI side chains Met-17 and Phe-34 create an exposed hydrophobic surface in place of Arg-17 and Val-34 in BPTI. The restriction this change places on protease target sequences is seen when the structure of APPI is superimposed on BPTI complexed to serine proteases, where the hydrophobic surface of APPI faces a complementary group of nonpolar side chains on kallikrein A versus polar side chains on trypsin.

Mouse placental receptor for basic somatomedin following maternal ethanol administration.

We have previously reported a reduction in the basic somatomedin (B-SM) binding activity of day-15 mouse placental membranes following 3 days of acute maternal ethanol administration. In the present experiments, we have investigated the effects of acute maternal ethanol administration early in gestation on the subsequent development of the placental B-SM receptor, and its relationship to alcohol-related embryofetal growth deficits. Following administration of aqueous ethanol (0.0, 3.6, 5.5 or 7.1 ml/kg) by gavage on days 7, 8 and 9 of gestation, there was no evidence of reproductive impairment in alcohol-treated dams, but there was a significant reduction in day-15 embryonic body weight. Crown-rump length was unaffected. There was no treatment-related difference in the percentage specific binding of [125I]-B-SM by day-15 placental membranes, or in maternal serum B-SM concentrations. These observations suggest that embryofetal growth restriction following maternal ethanol administration is not mediated directly by peripheral unresponsiveness of the B-SM receptor, at least when the exposure occurs early in or prior to placental receptor ontogeny.

The ontogeny of placental Na+-K+ ATPase in the mouse and its impairment by ethanol.

We have investigated the normal ontogeny of Na+-K+ ATPase in the mouse placenta and the possibility that impairment in placental transport capacity, as reflected in reductions in NA+-K+ ATPase activity, is associated with alcohol-related embryonic growth restriction. We have demonstrated that over the normal course of pregnancy there is a dramatic increase in placental NA+-K+ ATPase activity which occurs in concert with the embryofetal body growth spurt. Maternal ethanol administration during the early period of placental enzymogenesis (days 7-9) resulted in a significant reduction (up to 40%) of placental Na+-K+ ATPase activity on day 15. Both the severity and the frequency of the reduction were dose dependent. The effect was associated with significant reductions in embryonic body and brain weight but no change in body length or prenatal mortality. Incubation of term placental fragments for 2 h in increasing concentrations of ethanol resulted in a comparable reduction in enzyme activity. Our studies demonstrate that direct ethanol exposure produces a reduction of placental Na+-K+ ATPase activity, that exposure during the early stages of enzymogenesis results in persistent reductions in Na+-K+ ATPase activity in the mature placenta, and that this effect is associated with deficits of embryonic body and brain growth. A direct causal relationship has not been proven; however, it is conceivable that the correlation between reduced placental Na+-K+ ATPase activity and impaired embryofetal growth reflects a common causal pathway.

Multimodal liquid chromatography columns for the separation of proteins in either the anion-exchange or hydrophobic-interaction mode.

Several high-performance stationary phases suitable for protein chromatography were synthesized. Columns packed with these materials could be operated independently in either the anion-exchange or hydrophobic-interaction mode. Two approaches were used to prepare these materials. In the first method, a polyamine was adsorbed on the surface of macroporous silica and then crosslinked with a multifunctional oxirane. The hydrophobicity of the crosslinking agent and the extent of interconnection were used to modulate the electrostatic and solvophobic interactions. The second approach also utilized a crosslinked polyamine stationary phase; however, the forces of interaction were attenuated through controlled acylation of surface amines with a small anhydride molecule. The resolving ability of these columns, functioning in either mode, was comparable to commercial high-performance liquid chromatographic columns, designed to operate by a single retention mechanism. Column selectivity for proteins was completely different in each mode. Protein fractions collected from a multimodal column, operated in the anion-exchange mode, could be further purified by rechromatographing them on the same column in the hydrophobic-interaction mode. Utility of the multimodal column was demonstrated with the fractionation of several cytochromes and ferredoxins from the cyanobacterium Microcystis aeruginosa.

Ethanol neurotoxicity. 2. Direct effects on differentiating astrocytes.

In a previous investigation, we demonstrated altered patterns of growth, as indicated by RNA, DNA and protein contents, in rapidly growing astrocytes exposed to ethanol in primary culture. The present experiments were conducted to investigate the direct effects of ethanol on the differentiation of astrocytes as reflected in the activity of the astrocyte-specific enzyme, glutamine synthetase (Glu-S). Astroblasts obtained from the newborn mouse neopallium were continuously exposed to ethanol (0.0, 11.0, 22.3 or 44.5 mM) in primary cultures for 4, 11 or 18 days during the peak period of cell growth and differentiation. As seen previously, the lowest ethanol concentration (0.06 g/dl) had a "growth-promoting" effect on astroglia, reflected in an increase in protein content in ethanol-exposed cultures compared to controls. At higher concentrations (0.12 and 0.24 g/dl), there was a progressive "growth-impairing" effect. In contrast, the specific activity of Glu-S was reduced at all ethanol concentrations compared to controls in a concentration-dependent manner. By increasing the duration of exposure to ethanol, the effects on both protein content and Glu-S activity became more pronounced. It appears, however, that the timing of exposure relative to critical events in astrogliogenesis is a more important determinant of ethanol's toxicity than is duration of exposure. Derangements in astrocyte growth and differentiation may be major contributors to the pathogenesis of brain abnormalities in the fetal alcohol syndrome.