A Single Dose of Ibuprofen Impacts IL-10 Response to 164-km Road Cycling in the Heat.

Purpose: The purpose was to determine the effect of a single-dose prophylactic ibuprofen use before a 164-km road cycling event in high ambient temperature on the circulating cytokine and leukocyte responses. Methods: Twenty-three men (53 ± 8 y, 172.0 ± 22.0 cm, 85.1 ± 12.8 kg, 19.6 ± 4.4% body fat) completed a 164-km self-paced recreational road cycling event in a hot, humid, sunny environment (WBGT = 29.0 ± 2.9°C) after consuming 600 mg of ibuprofen (n = 13) or a placebo (n = 10). Blood samples were obtained one to two hours before (PRE) and immediately after (POST) the event, and analyzed for concentrations of circulating cytokines interleukins (IL)-1ß, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12, IL-13, GM-CSF, IFN-γ, and TNF-α and leukocytes (total leukocytes, granulocytes, monocytes, and lymphocytes). Results: Event completion time was 400.2 ± 74.8 min. Concentrations of all cytokines (except IL-1ß, IL-2, IL-5, IL-12, GM-CSF, and IFN-γ) and of all leukocyte subsets increased from PRE to POST. Ibuprofen ingestion attenuated the increase in IL-10 (86% increase with Ibuprofen; 270% increase with placebo). Conclusions: Consuming 600 mg of Ibuprofen prior to a 164-km road cycling event in a hot-humid environment attenuates exercise-induced increases in the concentration of the anti-inflammatory cytokine IL-10, but does not alter the effect of the exercise event on concentrations of other circulating cytokines or leukocyte subset concentrations.

Factors Associated with Pre-Event Hydration Status and Drinking Behavior of Middle-Aged Cyclists.

OBJECTIVES: Water is an essential nutrient for thermoregulation, metabolism, cognition, and overall physiological homeostatic function. However, aging adults display a blunted thirst mechanism and subsequently have an increased risk for dehydration or hyponatremia. Fluid consumption behaviors are modifiable and the importance of practicing adequate drinking behaviors for aging adults is amplified during exercise. Identification of aging adult's hydration beliefs and how they attain hydration advice could provide valuable information into ways to promote better drinking habits to reduce fluid imbalances. Thus, this investigation evaluated the knowledge, beliefs and behaviors of middle-aged cyclists (MA) that were associated with hydration status and drinking behavior, before and during a 164-km mass-participation event (ambient temperature, 33.3±2.8ºC(mean±SD)). DESIGN: This cross-sectional field study retrospectively grouped participants by their second urine specific gravity (Usg) measurement of the event morning prior to a mass participation cycling event. Usg was assessed via handheld refractometer. SETTING: The Hotter N' Hell Hundred 164-km cycling event in Wichita Falls, Texas during the month of August. PARTICIPANTS: 36 male recreational cyclists (age, 53±9 y(mean±SD)). MEASUREMENTS: Participants were grouped according their urine specific gravity as either slightly hyperhydrated (SH; n=12, Usg≤1.014), euhydrated (EUH; n=12, Usg, 1.015-1.020), or slightly dehydrated (SD; n=12, Usg≥1.021). Exercise histories and questionnaires were recorded 24-48 h prior to the cycling event. RESULTS: Regardless of pre-event hydration status, all groups experienced a similar body mass loss during the 164-km event and finished with statistically similar exercise times; also, drinking behavior within all groups was influenced by multiple factors. The primary factors associated with MA cyclist drinking behavior were trial and error/personal history and thirst; further, the majority of cyclists (≥65%) in SH, EUH, and SD believed that dehydration affects performance negatively. The least important factors included rehydration recommendations from scientific and sports medicine organizations, plus information from sports drink manufacturers. CONCLUSION: Considering the complexity of the present findings and the physiological changes that accompany aging such as delayed thirst perception, we recommend that MA cyclists formulate an individualized drinking plan that is based on observations during exercise.

Extracellular and cellular Hsp72 differ as biomarkers in acute exercise/environmental stress and recovery.

Stress-inducible Hsp72 is a potential biomarker to track risk of exertional heat illness during exercise/environmental stress. Characterization of extracellular (eHsp72) vs cellular Hsp72 (iHsp72) responses is required to define the appropriate use of Hsp72 as a reliable biomarker. In each of four repeat visits, participants (n = 6 men, 4 trials; total n = 24): (a) passively dehydrated overnight, (b) exercised (2 h) with no fluid in a hot, humid environmental chamber, (c) rested and rehydrated (1 h), (d) maximally exercised for 0.5 h, and (e) returned after 24 h of at-home recovery and rehydration. We measured rectal temperature, hydration status (% body mass loss, urine markers, serum osmolality), and Hsp72 (ELISA, flow cytometry. eHsp72 (circulating) and iHsp72 (CD3+ PBMCs) correlated (P < 0.05) with markers of heat, exercise, and dehydration stresses. eHsp72 immediately post-exercise (>15% above baseline, P < 0.05) decreased back to baseline levels by 1 h post-exercise, but iHsp72 expression continued to rise and remained elevated 24 h post-exercise (~2.5-fold baseline, P < 0.05). These data suggest that in addition to the classic physiological biomarkers of exercise heat stress, using cellular Hsp72 as an indicator of lasting effects of stress into recovery may be most appropriate for determining long-term effects of stress on risk for exertional heat illness.

RNA binding proteins regulate anabolic and catabolic gene expression in chondrocytes.

OBJECTIVE: Regulation of anabolic and catabolic factors is considered essential in maintaining the homoeostasis of healthy articular cartilage. In this study we investigated the influence of RNA binding proteins (RNABPs) in this process. DESIGN: Using small interfering RNA (siRNA), RNABP expression was knocked down in SW1353 chondrosarcoma cells and human articular chondrocytes. Gene expression and messenger RNA (mRNA) decay of anabolic (SOX9, Aggrecan) and catabolic (matrix metalloproteinase (MMP)13) factors were analysed using reverse transcription quantitative polymerase chain reaction (RT-qPCR). RNA-electromobility shift assays (EMSAs) were used to investigate RNABP interactions with the SOX9 mRNA 3' untranslated region (UTR). Immunohistochemical localisation of MMP13 and the RNABP human antigen R (HuR) was performed in E13.5 and E16.5 mouse embryo sections. RESULTS: SOX9 mRNA, mRNA half-life and protein expression were increased with siRNA targeting the RNABP tristetraprolin (TTP) in both HACs and SW1353s. TTP knockdown also stimulated aggrecan mRNA expression but did not affect its stability. RNA-EMSAs demonstrated that adenine uracil (AU)-rich elements in the SOX9 mRNA 3'UTR interacted with chondrocyte proteins with three specific elements interacting with TTP. HuR knockdown significantly increased MMP13 expression and also regulated the expression of a number of known transcriptional repressors of MMP13. HuR was ubiquitously expressed within mouse embryos yet displayed regional down-regulation within developing skeletal structures. CONCLUSION: This study demonstrates for the first time how RNABPs are able to affect the balance of anabolic and catabolic gene expression in human chondrocytes. The post-transcriptional mechanisms controlled by RNABPs present novel avenues of regulation and potential points of intervention for controlling the expression of SOX9 and MMP13 in chondrocytes.

Ageing does not result in a decline in cell synthetic activity in an injury prone tendon.

Advancing age is a well-known risk factor for tendon disease. Energy-storing tendons [e.g., human Achilles, equine superficial digital flexor tendon (SDFT)] are particularly vulnerable and it is thought that injury occurs following an accumulation of micro-damage in the extracellular matrix (ECM). Several authors suggest that age-related micro-damage accumulates due to a failure of the aging cell population to maintain the ECM or an imbalance between anabolic and catabolic pathways. We hypothesized that ageing results in a decreased ability of tendon cells to synthesize matrix components and matrix-degrading enzymes, resulting in a reduced turnover of the ECM and a decreased ability to repair micro-damage. The SDFT was collected from horses aged 3-30 years with no signs of tendon injury. Cell synthetic and degradative ability was assessed at the mRNA and protein levels. Telomere length was measured as an additional marker of cell ageing. There was no decrease in cellularity or relative telomere length with increasing age, and no decline in mRNA or protein levels for matrix proteins or degradative enzymes. The results suggest that the mechanism for age-related tendon deterioration is not due to reduced cellularity or a loss of synthetic functionality and that alternative mechanisms should be considered.

Comprehensive protein profiling of synovial fluid in osteoarthritis following protein equalization.

OBJECTIVE: The aim of the study was to characterise the protein complement of synovial fluid (SF) in health and osteoarthritis (OA) using liquid chromatography mass spectrometry (LC-MS/MS) following peptide-based depletion of high abundance proteins. DESIGN: SF was used from nine normal and nine OA Thoroughbred horses. Samples were analysed with LC-MS/MS using a NanoAcquity™ LC coupled to an LTQ Orbitrap Velos. In order to enrich the lower-abundance protein fractions protein equalisation was first undertaken using ProteoMiner™. Progenesis-QI™ LC-MS software was used for label-free quantification. In addition immunohistochemistry, western blotting and mRNA expression analysis was undertaken on selected joint tissues. RESULTS: The number of protein identifications was increased by 33% in the ProteoMiner™ treated SF compared to undepleted SF. A total of 764 proteins (462 with≥2 significant peptides) were identified in SF. A subset of 10 proteins were identified which were differentially expressed in OA SF. S100-A10, a calcium binding protein was upregulated in OA and validated with western blotting and immunohistochemistry. Several new OA specific peptide fragments (neopeptides) were identified. CONCLUSION: The protein equalisation method compressed the dynamic range of the synovial proteins identifying the most comprehensive SF proteome to date. A number of proteins were identified for the first time in SF which may be involved in the pathogenesis of OA. We identified a distinct set of proteins and neopeptides that may act as potential biomarkers to distinguish between normal and OA joints.

Cartilage damage involving extrusion of mineralisable matrix from the articular calcified cartilage and subchondral bone.

Arthropathy of the distal articular surfaces of the third metacarpal (Mc3) and metatarsal (Mt3) bones in the Thoroughbred racehorse (Tb) is a natural model of repetitive overload arthrosis. We describe a novel pathology that affects the articular calcified cartilage (ACC) and subchondral bone (SCB) and which is associated with hyaline articular cartilage degeneration. Parasagittal slices cut from the palmar quadrant of the distal condyles of the left Mc3/Mt3 of 39 trained Tbs euthanized for welfare reasons were imaged by point projection microradiography, and backscattered electron (BSE) scanning electron microscopy (SEM), light microscopy, and confocal scanning light microscopy. Mechanical properties were studied by nanoindentation. Data on the horses' training and racing career were also collected. Highly mineralised projections were observed extending from cracks in the ACC mineralising front into the hyaline articular cartilage (HAC) up to two-thirds the thickness of the HAC, and were associated with focal HAC surface fibrillation directly overlying their site. Nanoindentation identified this extruded matrix to be stiffer than any other mineralised phase in the specimen by a factor of two. The presence of projections was associated with a higher cartilage Mankin histology score (P<0.02) and increased amounts of gross cartilage loss pathologically on the condyle (P<0.02). Presence of projections was not significantly associated with: total number of racing seasons, age of horse, amount of earnings, number of days in training, total distance galloped in career, or presence of wear lines.

Longitudinal correlates of the persistence of irregular eating from age 5 to 14 years.

AIM: To report the stability of parent-perceived child irregular eating from 6 months to 14 years of age and to investigate a predictive model inclusive of child and parent factors. METHODS: Of the 7223 singleton children in a birth cohort, 5122 children were re-interviewed at 5 years and 4554 for the 14-year analysis. Information was obtained from structured interviews including questions answered by parents of the child at birth, 6 months, 5 years and 14 years; and by teenagers at age 14 years and from physical measures of the child. The mother's perception that the child was an irregular eater at age 14 years was the major outcome variable of interest. RESULTS: Approximately 40% of irregular eaters at age 5 will still be irregular eaters at age 14 years. This was not related to maternal education or socio-economic class. Significant at multivariate analysis were infant feeding problems and the children's ability to regulate their sleep and mood. Significant maternal factors were greater age, not feeling positive about the baby and persistent maternal anxiety during the child's early years. CONCLUSION: Irregular eating behaviour displays considerable continuity from childhood to mid-adolescence. Independent contributions to this behavioural phenotype include child biological and psychological factors and maternal anxiety during the child's early years.

Predictors of maternal misclassifications of their offspring's weight status: a longitudinal study.

BACKGROUND: Very little is known about the factors influencing parental misclassifications of a child's weight status. The aim of this study is to examine the predictors of maternal misclassifications of their adolescent offspring's weight status. METHODS: A mother-child linked analysis was carried out using 14-year follow-up data from a population-based prospective birth cohort of 2650 children (52% males) who were participants in the Mater-University Study of Pregnancy in Brisbane (Australia) in 1981. Offspring's observed height and weight and maternal perception of offspring weight were reported when they were 14 years old and predictors were prospectively recorded either at first clinical visit of mothers or at 5 or 14 years follow-up. Maternal misclassifications were defined combining observed body mass index (BMI) categories and maternal perceptions of their offspring's weight status. RESULTS: We found that maternal misclassification of child's weight status was common and included misclassifications both to higher and lower weight categories. Forty percent of mothers of overweight children misclassified their child as normal or underweight, more so in males than females. Fifteen percent of mothers of normal weight children misclassified their child as underweight, again more so in males than females. The main independent predictors of maternal misclassifications of child weight status were gender, child dissatisfaction with appearance, shape, size and weight, dieting to lose weight, general health status, maternal BMI and family meals. Gender, child dissatisfaction, dieting and maternal overweight were especially associated with misclassifications of overweight children. CONCLUSIONS: This study identified a number of maternal, child and family factors associated with maternal misclassifications of child weight status. Although relevant for clinical practice, further study is needed, however, to evaluate the benefits and harms of promoting increasing parental and child awareness of the child's weight status at a population level.

Intermedin (adrenomedullin-2): a novel counter-regulatory peptide in the cardiovascular and renal systems.

Intermedin (IMD) is a novel peptide related to calcitonin gene-related peptide (CGRP) and adrenomedullin (AM). Proteolytic processing of a larger precursor yields a series of biologically active C-terminal fragments, IMD(1-53), IMD(1-47) and IMD(8-47). IMD shares a family of receptors with AM and CGRP composed of a calcitonin-receptor like receptor (CALCRL) associated with one of three receptor activity modifying proteins (RAMP). Compared to CGRP, IMD is less potent at CGRP(1) receptors but more potent at AM(1) receptors and AM(2) receptors; compared to AM, IMD is more potent at CGRP(1) receptors but less potent at AM(1) and AM(2) receptors. The cellular and tissue distribution of IMD overlaps in some aspects with that of CGRP and AM but is distinct from both. IMD is present in neonatal but absent or expressed sparsely, in adult heart and vasculature and present at low levels in plasma. The prominent localization of IMD in hypothalamus and pituitary and in kidney is consistent with a physiological role in the central and peripheral regulation of the circulation and water-electrolyte homeostasis. IMD is a potent systemic and pulmonary vasodilator, influences regional blood flow and augments cardiac contractility. IMD protects myocardium from the deleterious effects of oxidative stress associated with ischaemia-reperfusion injury and exerts an anti-growth effect directly on cardiomyocytes to oppose the influence of hypertrophic stimuli. The robust increase in expression of the peptide in hypertrophied and ischaemic myocardium indicates an important protective role for IMD as an endogenous counter-regulatory peptide in the heart.

NPY and cardiac diseases.

Hypertension-induced left ventricular hypertrophy (LVH), along with ischemic heart disease, result in LV remodeling as part of a continuum that often leads to congestive heart failure. The neurohormonal model has been used to underpin many treatment strategies, but optimal outcomes have not been achieved. Neuropeptide Y (NPY) has emerged as an additional therapeutic target, ever since it was recognised as an important mediator released from sympathetic nerves in the heart, affecting coronary artery constriction and myocardial contraction. More recent interest has focused on the mitogenic and hypertrophic effects that are observed in endothelial and vascular smooth muscle cells, and cardiac myocytes. Of the six identified NPY receptor subtypes, Y(1), Y(2) and Y(5) appear to mediate the main functional responses in the heart. Plasma levels of NPY become elevated due to the increased sympathetic activation present in stress-related cardiac conditions. Also, NPY and Y receptor polymorphisms have been identified that may predispose individuals to increased risk of hypertension and cardiac complications. This review examines what understanding exists regarding the likely contribution of NPY to cardiac pathology. It appears that NPY may play a part in compensatory or detrimental remodeling of myocardial tissue subsequent to hemodynamic overload or myocardial infarction, and in angiogenic processes to regenerate myocardium after ischemic injury. However, greater mechanistic information is required in order to truly assess the potential for treatment of cardiac diseases using NPY-based drugs.

Isolation of E. coli O157:H7 from milk and cattle faeces from urban dairy farming and non dairy farming neighbour households in Dagoretti Division, Nairobi, Kenya: prevalence and risk factors.

OBJECTIVE: To estimate the prevalence of E. coli O157:H7 in milk and cattle faecal samples dairy and non dairy neighbouring households and to relate this prevalence to the risk to human health. DESIGN: Cross sectional study. SETTING: Urban and peri-urban households of Dagoretti, Division, Nairobi, Kenya. SUBJECTS: Dairy farming households and non dairy farming neighbouring households. RESULTS: E coli O157:H7 was isolated from milk samples at three of 136 non-dairy neighbour households (2.2% C.I. 0.5%,6.3%) but was not found in any of the milk samples from the 260 milk samples from dairy households (0% C.I. 0.0%,1.4%). E.coli O157:H7 was also found in fifteen of 285 pooled household cattle faecal sample (5.2%, C.I. 3.1%, 8.7%). One of the faecal isolates was found to have the marker for the production of VT1. Discussions with focus groups revealed that the participants had limited knowledge about E. coli O157:H7. Focus group discussions and household questionnaires revealed practices increasing risk of E. coli infections to humans are associated with milking hygiene, drinking water source and treatment, and manure handling. CONCLUSIONS: E. coli O157:H7 exists in urban setting and continuous surveillance is needed in case conditions and practices change favoring an increase in its prevalence and transmission to people.

Investigation into the prevalence of bovine cryptosporidiosis among small-holder dairy households in Dagoretti Division, Nairobi, Kenya.

OBJECTIVES: To determine the prevalence of bovine cryptosporidiosis and knowledge of this disease among smallholder dairy households in Dagoretti Division, Nairobi, Kenya. DESIGN: Cross-sectional survey of 299 dairy households and 149 non-dairy neighbouring households. SETTING: Dagoretti Division, a mixed urban and peri-urban area of Nairobi. SUBJECTS: Dairy householders and their immediate, non-cattle keeping neighbours. RESULTS: There were 50 positive households from 285 households where a fecal sample was collected giving an apparent prevalence of 18% [13, 23]%. Positive households were positively associated with watering cattle with well water, presence of shallow well, cemented cattle shed flooring and number of household cattle. (p < 0.05) Knowledge of cryptosporidiosis was generally low with more dairy respondents, 19% (56/292), who said they had heard of the disease than their non-dairy neighbours, 4% (6/146). CONCLUSION: This 18% bovine cryptosporidiosis prevalence does not constitute a risk to human health unless Cryptosporidium parvum is present. Typing of these positive samples is needed to assist in accurately determining the risk and enable evidence based management of urban dairy farming.

SB-271258, a major metabolite in the dog of the thiazolidinedione, rosiglitazone, stimulates protein synthesis in re-differentiated rat cardiomyocytes in vitro.

In dogs, chronic administration of thiazolidinediones causes cardiac hypertrophy in vivo at high doses. The hypertrophic action of rosiglitazone in dogs might be attributed to production of SB-271258 (a major metabolite in dogs but minor in rats and man), rather than a direct effect on myocardium. The hypothesis that SB-271258 had potential to initiate cardiomyocyte hypertrophy or to modify responses elicited by other hypertrophic stimuli was tested in an in vitro bioassay utilising adult rat ventricular cardiomyocytes. SB-271258 increased protein and incorporation of 14C-phenylalanine, a marker of protein synthesis, in rat cardiomyocytes maintained in serum-free culture (24 h), maximally at 1 microM by 15.0% and 9.1%, respectively. In the presence of serum (10% v/v), SB-271258 elicited a moderate trophic effect: cellular protein and incorporation of 14C-phenylalanine were increased, maximally at 100 nM by 31.7% and 36.3%, respectively, above basal values (18.6% and 13.3% increases above serum response). In the presence of IGF-1 (10 nM) plus SB-271258, protein synthesis was increased, maximally by 45.5% above basal value (increase of 6.9% above IGF-1 alone). In contrast, SB-271258 attenuated the increase (12.0%) in cellular protein elicited by IGF-1. In re-differentiated cardiomyocytes, a model of relevance to established hypertrophy, SB-271258 (1 nM-1 microM) elicited a marked trophic effect per se, as evidenced by the maximum increase (at 100 nM), in protein synthesis of 24.5%. In conclusion, these data imply that cardiac hypertrophy associated with chronic administration of rosiglitazone in dogs in previous in vivo studies might be partly attributable to production of the metabolite SB-271258 since this metabolite was shown to elicit trophic effects directly on rat cardiomyocytes.

Phenol red thread test vs Schirmer's test: a comparative study.

AIM: To measure the agreement between Schirmer's and phenol red thread tests in detecting dry eyes. PATIENTS AND METHODS: A total of 103 patients attending preoperative cataract assessment clinic who agreed to be involved in the study were recruited. Each patient had one eye examined by both tests in a random order by two different investigators who were unaware of the results of the other test. Dry eye symptoms were assessed using an interviewer-administered questionnaire. The data were collected after the study period and analysed using kappa statistics to assess the agreement between the two tests. RESULTS: Schirmer's test was positive in 25 patients when a cutoff point of 5 mm was used and positive in 41 patients with a cutoff point of 10 mm. Phenol red thread test was positive in four patients when a cutoff point of 10 mm was used and in 32 patients with a cutoff point of 20 mm. Kappa ranged from 0.067 to 0.3 indicating very weak agreement between the two tests. In all, 27% of the patients had symptoms of dry eyes; however, the agreement between each test and the symptoms was very poor. CONCLUSION: There is very weak agreement between Schirmer's test and phenol red thread tests and between each test and symptoms of dry eyes.

Eating disorders and oral health: a review of the literature.

This article is a review of the recent literature pertaining to the oral sequelae of eating disorders (EDs). Dentists are recognized as being some of the first health care professionals to whom a previously undiagnosed eating disorder patient (EDP) may present. However, despite the prevalence (up to 4 per cent) of such conditions in teenage girls and young adult females, there is relatively little published in the recent literature regarding the oral sequelae of EDs. This compares unfavourably with the attention given recently in the dental literature to conditions such as diabetes mellitus, which have a similar prevalence in the adult population. The incidence of EDs is increasing and it would be expected that dentists who treat patients in the affected age groups would encounter more individuals exhibiting EDs. Most of the reports in the literature concentrate on the obvious clinical features of dental destruction (perimolysis), parotid swelling and biochemical abnormalities particularly related to salivary and pancreatic amylase. However, there is no consistency in explanation of the oral phenomena and epiphenomena seen in EDs. Many EDPs are nutritionally challenged; there is a relative lack of information pertaining to non-dental, oral lesions associated with nutritional deficiencies.