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Povidone-iodine (Polidine) is a synthetic broad-spectrum antiseptic and being applied topically to treat wounds and prevent their infection. It is however used by poultry farmers, field veterinarians, and other animal health workers with the claim that it is effective for treatment of infectious bursal disease when administered orally. Hence, an acute oral toxicity study was conducted to ascertain its safety profile. Ten cockerel chicks were randomly selected and divided into 2 groups of 5 chicks per group. One group served as the negative control, whereas the other group was administered povidone-iodine at a dose of 2,000 mg/kg of BW orally. The blood sample was collected at the end of the study to determine changes in hematological and biochemical parameters. In addition, vital organs were also harvested and preserved for histopathological examinations. The result showed that the median lethal dose (LD50) of the povidone-iodine is higher than 2,000 mg/kg of BW in cockerels. There were no significant changes in the hematological parameters measured. Biochemical evaluation (renal and liver function test) showed an increase in aspartate aminotransferase, alanine aminotransferase, and alkaline phosphatase levels after administration of povidone-iodine. The study indicated that the LD50 of povidone-iodine is higher than 2,000 mg/kg of BW of cockerels, and there were increases in urinary and liver enzymes at this dose.

This study was conducted to evaluate potential hormonal mechanisms associated with the stress response, thermoregulation, and metabolic changes of broiler chickens exposed to high environmental temperature. Nine hundred 1-day-old male broiler chicks (Ross 708) were placed in floor pens and raised to 24 d. At 24 d, chicks were randomly assigned to 1 of 2 treatments, heat stress (HS) or no HS, and allocated into battery cages in 8 batteries (10 birds per cage, 2 cages per battery). On day 31, blood was collected prior to HS and analyzed using an iSTAT analyzer. Half of the batteries were then moved into 2 rooms with an elevated ambient temperature (35°C) for 8 h. The remaining batteries stayed in the thermoneutral rooms with an ambient temperature of 22°C. Beginning at 5 h after the initiation of HS, blood was collected and analyzed using an iSTAT analyzer, birds were euthanized, and hypothalamus and pituitary samples were collected (16 birds per treatment), flash frozen, and stored at -80°C until RNA extraction. Reverse transcription-quantitative PCR was used to compare mRNA levels of key corticotropic and thyrotrophic genes in the hypothalamus and pituitary. Levels of mRNA for each target gene were normalized to PGK1 (pituitary) and GAPDH (hypothalamus) mRNA. Differences were determined using mixed model ANOVA. HS decreased (P < 0.05) feed intake, BW, bicarbonate, potassium, CO2, and triiodothyronine, while it increased mortality, glucose, pH, plasma thyroxine, and corticosterone. Expression of pituitary corticotropin-releasing hormone receptor 1 was downregulated (P < 0.001), while corticotropin-releasing hormone receptor 2 mRNA levels were higher (P = 0.001) in HS birds. HS increased expression of thyroid hormone receptor β (P = 0.01) (2.8-fold) and thyroid stimulating hormone β (P = 0.009) (1.4-fold). HS did not affect levels of mRNA of genes evaluated in the hypothalamus. Results showed that HS significantly affected both the thyrotropic and corticotropic axes. Understanding the role and regulation of these pathways during HS will allow researchers to better evaluate management strategies to combat HS.

A total of 960 male Cobb 500 broilers were used in a growth performance study to explore the effect of coccidial vaccination and/or coccidial challenge on blood biochemistry and veterinary postmortem metrics. Day-old chicks were randomly divided into one of the 4 experimental treatments. Treatments were arranged in a 2 × 2 factorial arrangement, with the factors being without or with vaccination (administered on day 1) or coccidial challenge (oral gavage on day 7). Growth performance was monitored on a weekly basis. Blood sample collection, as well as full veterinary necropsies, were carried out on days 6, 8, 13, 20, 27, and 34. Birds that did not receive the vaccination but were challenged with coccidiosis had higher feed conversion ratio, lower body weights, and higher mortality than the other experimental groups, and this effect was particularly evident from day 13 to day 20. Birds challenged with coccidiosis had lower plasma sodium and total carotenoid concentrations and higher potassium and globulin concentrations than nonchallenged birds. Significant interactions between age and experimental treatment for these blood parameters were observed, particularly on day 13. The necropsy results confirmed the effectiveness of the challenge and vaccination treatments, wherein birds that were challenged had higher coccidiosis scores on day 13 and day 27 than birds that were not challenged. These results demonstrate the potential for plasma sodium, potassium, total protein, total carbon dioxide, globulin, and carotenoid analysis for early diagnosis of coccidiosis in growing broiler chickens. Further work is necessary to establish whether the changes in blood biochemistry observed in the present study are transferable to alternative flocks of chicken and whether early diagnosis and intervention may mitigate performance losses associated with this disease.

International audience; Apolipoproteins govern lipoprotein metabolism and are promising biomarkers of metabolic and cardiovascular diseases. Unlike immunoassays, MS enables the quantification and phenotyping of multiple apolipoproteins. Hence, here, we aimed to develop a LC-MS/MS assay that can simultaneously quantitate 18 human apolipoproteins [A-I, A-II, A-IV, A-V, B48, B100, C-I, C-II, C-III, C-IV, D, E, F, H, J, L1, M, and (a)] and determined apoE, apoL1, and apo(a) phenotypes in human plasma and serum samples. The plasma and serum apolipoproteins were trypsin digested through an optimized procedure and peptides were extracted and analyzed by LC-MS/MS. The method was validated according to standard guidelines in samples spiked with known peptide amounts. The LC-MS/MS results were compared with those obtained with other techniques, and reproducibility, dilution effects, and stabilities were also assessed. Peptide markers were successfully selected for targeted apolipoprotein quantification and phenotyping. After optimization, the assay was validated for linearity, lower limits of quantification, accuracy (biases: -14.8% to 12.1%), intra-assay variability [coefficients of variation (CVs): 1.5-14.2%], and inter-assay repeatability (CVs: 4.1-14.3%). Bland-Altman plots indicated no major statistically significant differences between LC-MS/MS and other techniques. The LC-MS/MS results were reproducible over five repeated experiments (CVs: 1.8-13.7%), and we identified marked differences among the plasma and serum samples. The LC-MS/MS assay developed here is rapid, requires only small sampling volumes, and incurs reasonable costs, thus making it amenable for a wide range of studies of apolipoprotein metabolism. We also highlight how this assay can be implemented in laboratories.

The present work was carried out to investigate the influences of housing system and dietary essential oils (EOs) supplementation to laying hens on the productive performance, egg quality, immunity, antioxidant parameters, and hematology. A factorial arrangement (2 × 4) was performed, including 2 housing systems and 4 different types of EOs (without EOs, thymol, carvacrol, and euganol) during the production stages (from 28–78 wk of age). Birds were randomly divided into 2 groups with each of 2,000 birds. The first group was moved to laying cages while the second group was a floor reared. Each group was randomly divided into 4 groups (5 replicates of 100 birds each): The first were considered as a control group, and the second, third, and fourth groups were treated with thymol, carvacrol, and euganol EO, respectively. The results showed that hens reared in cage system had higher egg weight (P 0.05) by housing system. The groups fed EOs showed a rapid improvement (P

This study was performed to investigate the effects of lignocellulose supplementation (LS) on performance parameters, egg quality, aerobic bacterial load of eggshell, serum biochemical parameters, and jejunal histomorphological traits of laying hens between 18 and 38 wk of age. A total of 640 pullets at 16 wk of age were allotted to 4 treatment groups as 0 kg (control, CONT), 0.5 kg, 1 kg, and 2 kg LS per ton of feed. Body weight (BW), daily feed intake, egg production (EP), egg weight (EW), and efficiency of feed utilization (EF) were determined as the mean of each 3-wk period between 18 and 38 wk of age. Laying hens in the 1 kg LS group had a higher BW mean (1632.1 g, P < 0.001). the highest mean value of EP and EW were observed in 1 kg LS group (81.8% and 57.3 g, respectively), whereas the lowest values were found in the 2 kg LS group (78.6% and 54.4 g, respectively, P < 0.001). the mean of EF was the lowest in the 1 kg LS group (2.72, P < 0.001). There was a decline in eggshell breaking strength and eggshell thickness in the 2 kg LS, when compared with the 0.5 and 1 kg LS groups (P < 0.001). the total aerobic bacterial load of the eggshell was the lowest in the 1 kg LS group (4.7 log10 cfu/mL). the level of aspartate amino transferase and alanine amino transferase showed an incre-ment in both the CONT and 2 kg LS groups (P < 0.001). the high level of LS (2 kg per ton of feed) caused a decline in the levels of IgY, IgA and IgM, when compared to the 0.5 and 1 kg LS groups (P < 0.001). Laying hens in 0.5 and 1 kg LS groups had longer villus height (1335.9 mm) in the jejunum than the others (P < 0.001). These find-ings showed that the 1 kg LS per ton of feed improved EP and EW, eggshell quality, immunoglobulin levels and intestinal morphology, and decreased the total aerobic bacterial load., Global Nutritech Biotechnology LLC (Richmond, VA, USA), The authors would like to thank Global Nutritech Biotechnology LLC (Richmond, VA, USA) for their financial support.

Less evidence is available currently to reveal whether the immune system and productivity of laying hens change under long periods of ammonia exposure in hot climate. The present study was conducted to determine the effects of chronic exposure to high temperature and ammonia concentrations on health, immune response, and reproductive hormones of commercial laying hens. A total of five hundred and seventy six 20-week-old laying hens (Hy-Line Brown) were used in this study. Birds were housed in cages (4 birds per cage) and received 16-wk treatments in 6 artificial environmental chambers. Hens were allocated to 6 treatments: treatment 1 (T1, 20°C, ≤5 ppm, control group), treatment 2 (T2, 20°C, 20 ppm), treatment 3 (T3, 20°C, 45 ppm), treatment 4 (T4, 35°C, ≤5 ppm), treatment 5 (T5, 35°C, 20 ppm), and treatment 6 (T6, 35°C, 45 ppm). Blood samples were collected at 22, 26, 30, 34, and 38 wk of age and plasma IgG, IgM, IgA, corticosterone (CORT), total antioxidant capacity (T-AOC), luteinizing hormone (LH), estradiol (E2), and follicular stimulating hormone (FSH) were measured. The results of this study showed that high ambient temperature and excessive ammonia increased the concentration of IgG but decreased the concentration of IgA, T-AOC, LH, FSH, and E2 of hens compared with those of the control birds. From the age of 34 wk, significantly increased concentrations of IgG were observed in hens exposed to moderate and high levels of ammonia. CORT level showed marked differences between the treatments only at the age of 26 wk. In addition, LH and E2 of hens demonstrated significant differences among the treatments in the middle and later stages of the experiment, while FSH levels of the control birds were significantly higher than the others at the age of 38 wk. Excessive ammonia in high temperature was a physiological stress factor that had a negative effect, which inhibited immune function and impacted the reproductive hormones.

The phenotype of modern commercial turkeys is substantially different than that of unselected, heritage turkey lines. These phenotypic changes have arisen from alterations in the genome/transcriptome, as well as the influence of many external factors on growth performance including nutrition, environment, and management. To investigate the phenotypic changes resulting from genetic selection for increased body weight, The Ohio State University maintains 2 unique genetic turkey lines: the randombred control (RBC2) line, which is comprised of genetics from 1960 era commercial turkeys and has been maintained without conscious selection for any trait; and the F line, which was originally selected from the RBC2 line and has been selected for increased 16 wk body weight for over 50 generations. This study used broad-spectrum mass-spectrometry profiling techniques to identify and quantify differences in the metabolome of the serum of F and RBC2 turkey lines. Serum samples from both F and RBC2 turkeys were subject to quantitative time of flight liquid chromatography tandem mass spectrometry analyses. Principle component analyses showed distinct populations of metabolites in the F vs. RBC2 serum, suggesting that increased body weight is associated with the accumulation of several metabolites. Comparing the spectral features to online databases resulted in the selection of 104 features with potentially identifiable chemical structures. Of these 104 features, 25 were found at higher levels in the serum of the RBC2 line turkeys, while 79 were found at a greater abundance in the F line turkeys. A more detailed analysis of these 104 features allowed for the putative identification of 49 compounds, which were clustered into 6 functional groups: 1) energy metabolism; 2) vitamins; 3) hormones and signaling molecules; 4) lipid derivatives, fatty acid metabolites, and membrane components; 5) amino acid/protein metabolism; and 6) microbial metabolites. Further validation and experimentation is needed to confirm the identity of these metabolites and understand their biological relevance and association with selection for increased body weight.

This study evaluated the effects of different dietary zinc (Zn) levels on productive and reproductive performance, egg quality, tibial characteristics, plasma biochemical and antioxidant indices, and zinc deposition in laying duck breeders. A total of 504 Longyan duck breeders aged 21 wk were randomly allocated to 6 treatments and fed a basal diet (Zn, 27.7 mg/kg) or that basal diet supplemented with Zn (as ZnSO4·H2 O) at 10, 20, 40, 80, or 160 mg Zn per kg of feed for 20 wk. Each group had 6 replicates of 14 ducks each. Dietary Zn supplementation affected (P < 0.05) the egg production, FCR, and shell thickness of laying duck breeders from 21 to 40 wk, and there was a quadratic (P < 0.05) effect between them. Dietary Zn supplementation affected (P < 0.05) and quadratically (P < 0.001) increased the breaking strength, density, and dry defatted weight of tibias. Alkaline phosphatase, calcium, phosphorus, total superoxide dismutase, glutathione peroxidase (GSH-Px), and malondialdehyde (MDA) activities or content in plasma were affected (P < 0.05), and quadratically (P < 0.01) changed by dietary Zn levels. Dietary Zn supplementation affected (P < 0.01) and increased the Zn deposition in egg yolk (linear, P < 0.05; quadratic, P < 0.001) and tibia (linear, P < 0.05). The dietary Zn requirements, in mg/kg for a basal diet containing 27.7 mg/kg Zn, for Longyan duck breeders from 21 to 40 wk of age were estimated to be 65.4 for optimizing egg production, 68.6 for FCR, 102 for hatchling BW, 94.7 for eggshell thickness, 77.2 for tibial breaking strength, 81.4 for tibial density, 78.9 for tibial dry defatted weight, 69.5 for plasma GSH-Px activity, 72.4 for plasma MDA content, and 94.6 for Zn content in tibia. Overall, dietary Zn supplementation, up to 160 mg/kg feed, affected the productive performance, eggshell thickness, tibial characteristics, plasma antioxidant status, and Zn deposition of layer duck breeders. Supplementing this basal diet (27.7 mg/kg Zn) with 70 to 80 mg/kg additional Zn was adequate for laying duck breeders during the laying period.

Blood tests, which are used to evaluate health status, are relatively non-invasive and provide a great deal of health-related information. Blood is usually collected using a standard venous blood sampling protocol, but it is possible to collect blood from a subject's fingertip, and previous studies have investigated whether fingertip-derived blood can be used for various blood tests. In this study, the proteomes and metabolomes of venous and fingertip plasma were analyzed using non-targeted proteomics and metabolomics, respectively. In proteomics, the levels of 523 proteins were compared between venous and fingertip plasma. The correlation coefficient (r) for the relationship between protein levels of venous and fingertip plasma was 0.9999. Some proteins had high fingertip to venous plasma level ratios (finger:venous ratios), whereas others had low finger:venous ratios, and the mean±standard deviation (SD) finger:venous ratio was 0.994 ± 0.304. In metabolomics, 40, 33, and 216 cationic metabolites, anionic metabolites, and lipids, respectively, were detected in venous plasma, and the equivalent figures for fingertip plasma were 40, 35, and 216, respectively. Regarding the correlations between metabolite levels in venous and fingertip plasma, the correlation coefficients (r) for cationic metabolites, anionic metabolites, and lipids were 0.9952, 0.9699, and 0.9980, respectively. The mean±SD finger:venous ratio was 1.19 ± 0.584 for cationic metabolites, 1.23 ± 0.548 for anionic metabolites, and 1.00 ± 0.245 for lipids. Our study suggests that it might be possible to use fingertip plasma to measure plasma protein and metabolite levels, and will contribute to development of a fingertip blood sampling procedure for measuring blood biomarker levels.

Native myocardial T1 and extracellular volume (ECV) by cardiovascular magnetic resonance imaging are key biomarkers for assessing several heart muscle diseases. Their measurement, however, is influenced by blood T1, either due to myocardial vascular blood volume or via blood pool partial voluming.

In many mammalian species, corpus luteum derived progesterone (P4) is the main functional gestagen during the estrous cycle and pregnancy. P4 can be metabolized into various metabolites, of which some are biologically active. While some metabolites target the classical nuclear progesterone receptor (PR), neurosteroids bind the receptors of type A γ-aminobutyric acid (GABAA-r) in the brain. According to the position of reduction within the molecule, metabolites of P4 can be characterized into C20-reduced progestogens (20α-dihydroprogesterone (20α-DHP) and 20β-dihydroprogesterone (20β-DHP)), C3-reduced progestogens (3α-dihydroprogesterone (3α-DHP) and 3β-dihydroprogesterone (3β-DHP)), 5α-reduced progestogens (5α-dihydroprogesterone (5α-DHP), allopregnanolone and isopregnanolone) and 5β-reduced progestogens (5β-dihydroprogesterone (5β-DHP), pregnanolone and epipregnanolone). We questioned whether the reduced progestogens are present in bovine plasma during the estrous cycle and whether their profiles differed from the profile of the common precursor P4 around the time of luteolysis. The analytes were monitored in plasma samples using liquid chromatography mass spectrometry (LC-MS). While progestogens lagged behind the drop of P4 at luteolysis, they followed the profile of P4 during the estrous cycle. The abundance of P4 was predominant followed by allopregnanolone, pregnanolone, epipregnanolone and 20β-DHP. Further studies will need to focus particularly on the period around luteolysis., Theriogenology, 142, ISSN:1879-3231, ISSN:0093-691x

One of the most important causes of the high mortality rate and low life expectancy of lung cancer is the detection at advanced stages. Thus, there is an urgent need for early diagnosis and the search of new selective biomarkers. Selenium is an important constituent of selenoproteins and a powerful antioxidant able to protect against cancer. In this work, the absolute quantification of selenium in selenoproteins and the total content in selenometabolites has been performed for the first time in serum from lung cancer patients (LC) and healthy controls (HC). To this end, a method for the simultaneous speciation of selenoproteins using size exclusion chromatography (SEC) and affinity chromatography (AF) with detection by ICP-QQQ-MS, and quantification by isotopic dilution (IDA) (SEC-AF-HPLC-SUID-ICP-QQQ-MS) was developed to determine the selenium concentration in eGPx, SEPP1 and SeAlb, as well as total selenometabolites, to find alterations that may serve as biomarkers of this disease. In the same way, a method based on anion-exchange chromatography coupled to ICP-QQQ-MS was developed to quantify selenometabolites (SeCys2, SeMeSeCys, SeMet, selenite and selenate) in the same LC and HC serum samples. The results showed that the averaged concentrations of selenium in eGPx, SeAlb and selenite were significantly higher in LC patients (LC (eGPx: 21.24 +/- 0.77 ng g(-1); SeAlb: 49.56 +/- 3.16 ng g(-1) and Se(IV): 6.20 +/- 1.22 ng g(-1)) than in HC group (eGPx: 16.96 +/- 0.53 ng g(-1); SeAlb: 38.33 +/- 2.66 ng g(-1) and Se(IV): 3.56 +/- 0.55 ng g(-1)). In addition, the ratios between selenoproteins and selenometabolites have been calculated for the first to study their potential use as LC biomarkers. The rates eGPx/SEPP1, SEPP1/SeAlb, eGPx/Se(IV) and SEPP1/Se(IV) were significantly different between LC and HC groups. (C) 2020 Elsevier B.V. All rights reserved This research was funded by the projects PGC2018-096608-B-C21 from the Spanish Ministry of Economy and Competitiveness, and P12-FQM-0442 from the Regional Ministry of Economy, Innovation, Science and Employment (Andalusian Government, Spain). Projects Neumosur (8/2012 and 9/2015) and SEPAR (124/2012 and 091/2016). Finally, authors are grateful to FEDER (European Community) for financial support, grants number UNHU13-1E-1611 and UNHU15-CE-3140.

Human blood is a self-regenerating lipid-rich biological fluid that is routinely collected in hospital settings. The inventory of lipid molecules found in blood plasma (plasma lipidome) offers insights into individual metabolism and physiology in health and disease. Disturbances in the plasma lipidome also occur in conditions that are not directly linked to lipid metabolism; therefore, plasma lipidomics based on MS is an emerging tool in an array of clinical diagnostics and disease management. However, challenges exist in the translation of such lipidomic data to clinical applications. These relate to the reproducibility, accuracy, and precision of lipid quantitation, study design, sample handling, and data sharing. This position paper emerged from a workshop that initiated a community-led process to elaborate and define a set of generally accepted guidelines for quantitative MS-based lipidomics of blood plasma or serum, with harmonization of data acquired on different instrumentation platforms across independent laboratories as an ultimate goal. We hope that other fields may benefit from and follow such a precedent.

A targeted quantitative metabolomics approach was used to study temporal changes of serum metabolites in cows that normally released their fetal membranes and those that retained the placenta. We identified and measured serum concentrations of 128 metabolites including amino acids, acylcarnitines, biogenic amines, glycerophospholipids, sphingolipids and hexose at -8 and -4 weeks before parturition, during the week of retained placenta (RP) diagnosis, and at +4 and +8 weeks after parturition. In addition, we aimed at identifying metabolite signatures of pre-RP in the serum that might be used as predictive biomarkers for risk of developing RP in dairy cows. Results revealed major alterations in the metabolite fingerprints of pre-RP cows starting as early as -8 weeks before parturition and continuing as far as +8 weeks after calving. Biomarker candidates found in this study are mainly biomarkers of inflammation which might not be specific to RP. Therefore, the relevance of serum Lys, Orn, acetylornithine, lysophophatidylcholine LysoPC a C28:0, Asp, Leu and Ile as potential serum biomarkers for prediction of risk of RP in dairy cows will have to be tested in the future. In addition, lower concentrations of LysoPCs, Trp, and higher kynurenine in the serum during prepartum and the week of occurrence of RP suggest involvement of inflammation in the pathobiology of RP.

Made available in DSpace on 2018-11-26T16:28:07Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-01-01 Intramural Research Program of the Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health National Institute for Health Research Background: A common challenge in medicine, exemplified in the analysis of biomarker data, is that large studies are needed for sufficient statistical power. Often, this may only be achievable by aggregating multiple cohorts. However, different studies may use disparate platforms for laboratory analysis, which can hinder merging. Methods: Using circulating placental growth factor (PIGF), a potential biomarker for hypertensive disorders of pregnancy (HDP) such as preeclampsia, as an example, we investigated how such issues can be overcome by inter-platform standardization and merging algorithms. We studied 16,462 pregnancies from 22 study cohorts. PIGF measurements (gestational age >= 20 weeks) analyzed on one of four platforms: R & Systems, Alere (R) Triage, Roche (R) Elecsys or Abbott (R) Architect, were available for 13,429 women. Two merging algorithms, using Z-Score and Multiple of Median transformations, were applied. Results: Best reference curves (BRC), based on merged, transformed PIGF measurements in uncomplicated pregnancy across six gestational age groups, were estimated. Identification of HDP by these PIGF-BRCS was compared to that of platform-specific curves. Conclusions: We demonstrate the feasibility of merging PIGF concentrations from different analytical platforms. Overall BRC identification of HDP performed at least as well as platform-specific curves. Our method can be extended to any set of biomarkers obtained from different laboratory platforms in any field. Merged biomarker data from multiple studies will improve statistical power and enlarge our understanding of the pathophysiology and management of medical syndromes. (C) 2015 International Society for the Study of Hypertension in Pregnancy. Published by Elsevier B.V. All rights reserved. Univ Oxford, Nuffield Dept Populat Hlth, Old Rd Campus, Oxford OX3 7LF, England Univ British Columbia, Dept Obstet & Gynaecol, Vancouver, BC V5Z 1M9, Canada Univ British Columbia, Child & Family Res Inst, Vancouver, BC V5Z 1M9, Canada Univ Oxford, Nuffield Dept Obstet & Gynaecol, Oxford, England Yale Univ, New Haven, CT USA Univ Milan, I-20122 Milan, Italy Kings Coll London, Womens Hlth Acad Ctr, London WC2R 2LS, England Univ Barcelona, Hosp Clin, Barcelona, Spain Hosp Univ 12 Octubre, Dept Obstet & Gynecol, Fetal Med Unit, Madrid, Spain Univ Complutense, E-28040 Madrid, Spain Michigan State Univ, E Lansing, MI 48824 USA Univ Pittsburgh, Magee Womens Res Inst, Pittsburgh, PA USA Aarhus Univ Hosp, Dept Obstet & Gynecol, DK-8000 Aarhus, Denmark Aarhus Univ, Aarhus, Denmark Aarhus Univ Hosp, Dept Oncol, DK-8000 Aarhus, Denmark Univ Helsinki, Med Genet Obstet & Gynaecol, Helsinki, Finland Univ Helsinki, Inst Mol Med Finland, Helsinki, Finland Helsinki Univ Hosp, Helsinki, Finland Univ Basel, Basel, Switzerland Harvard Univ, Brigham & Womens Hosp, Sch Med, Boston, MA 02115 USA Klagenfurt & Med Univ, Interdisciplinary Ctr Gynecol Gynecooncol & Fetom, Graz, Austria St Marys Hosp, Maternal & Fetal Hlth Res Ctr, Manchester M13 0JH, Lancs, England Univ Manchester, Manchester, Lancs, England Univ Texas Houston, Sch Publ Hlth, Houston, TX USA Univ Estadual Paulista, Dept Obstet, Botucatu, SP, Brazil Univ Texas Med Branch, Galveston, TX 77555 USA Univ Amsterdam, Acad Med Ctr, Dept Obstet & Gynaecol, Meibergdreef 9, NL-1105 AZ Amsterdam, Netherlands Erasmus MC, Rotterdam, Netherlands Univ Jena, D-07745 Jena, Germany Univ Leipzig, D-04109 Leipzig, Germany Univ Paris 05, Paris, France Charite, D-13353 Berlin, Germany Univ Helsinki, Obstet & Gynecol, Helsinki, Finland NIHR Univ Coll London Hosp, Biomed Res Ctr, Inst Womens Hlth, London, England Med Univ Vienna, Dept Obstet & Gynecol, Vienna, Austria Oslo Univ Hosp, Dept Obstet & Gynecol, Oslo, Norway Univ Oslo, Oslo, Norway Univ Estadual Paulista, Dept Obstet, Botucatu, SP, Brazil National Institute for Health Research: RP-2014-05-019

Residual feed intake (RFI), defined as the difference between an animal's actual feed intake and expected feed intake over a specific period, is an inheritable character of feed conversion efficiency in dairy cows. Research has shown that a lower RFI could improve the profitability of milk production. This study explored variation in RFI by comparing the differences in body size, milk performance, feeding behavior, and serum metabolites in 29 Holstein cows in mid lactation. The cows were selected from a total of 84 animals based on their RFI following feedlot tests. Selected cows were ranked into high RFI (RFI >1 SD above the mean, n=14) and low RFI (RFI

Rationale & Objective Chronic kidney disease (CKD) is complicated by abnormalities that reflect disruption in filtration, tubular, and endocrine functions of the kidney. Our aim was to explore the relationship of specific laboratory result abnormalities and hypertension with the estimated glomerular filtration rate (eGFR) and albuminuria CKD staging framework. Study Design Cross-sectional individual participant-level analyses in a global consortium. Setting & Study Populations 17 CKD and 38 general population and high-risk cohorts. Selection Criteria for Studies Cohorts in the CKD Prognosis Consortium with data for eGFR and albuminuria, as well as a measurement of hemoglobin, bicarbonate, phosphorus, parathyroid hormone, potassium, or calcium, or hypertension. Data Extraction Data were obtained and analyzed between July 2015 and January 2018. Analytical Approach We modeled the association of eGFR and albuminuria with hemoglobin, bicarbonate, phosphorus, parathyroid hormone, potassium, and calcium values using linear regression and with hypertension and categorical definitions of each abnormality using logistic regression. Results were pooled using random-effects meta-analyses. Results The CKD cohorts (n = 254,666 participants) were 27% women and 10% black, with a mean age of 69 (SD, 12) years. The general population/high-risk cohorts (n = 1,758,334) were 50% women and 2% black, with a mean age of 50 (16) years. There was a strong graded association between lower eGFR and all laboratory result abnormalities (ORs ranging from 3.27 [95% CI, 2.68-3.97] to 8.91 [95% CI, 7.22-10.99] comparing eGFRs of 15 to 29 with eGFRs of 45 to 59 mL/min/1.73 m 2), whereas albuminuria had equivocal or weak associations with abnormalities (ORs ranging from 0.77 [95% CI, 0.60-0.99] to 1.92 [95% CI, 1.65-2.24] comparing urinary albumin-creatinine ratio > 300 vs < 30 mg/g). Limitations Variations in study era, health care delivery system, typical diet, and laboratory assays. Conclusions Lower eGFR was strongly associated with higher odds of multiple laboratory result abnormalities. Knowledge of risk associations might help guide management in the heterogeneous group of patients with CKD. © 2018. This is the authors’ accepted and refereed manuscript to the article. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/ "

The ultimate goal of an artificial pancreas is finding the optimal insulin rates that can effectively reduce high blood glucose (BG) levels in type 1 diabetic patients. To achieve this, most closed-loop control strategies need to compute the optimal insulin action on the basis of precedent glucose and insulin levels. Unlike glucose levels which can be measured in real-time, unavailability of insulin sensors makes it essential the use of mathematical models to estimate plasma insulin concentrations. Between others, filtering techniques based on a generalization of the Kalman filter (KF) have been the most widely applied in the estimation of hidden states in nonlinear dynamic systems. Nevertheless, poor predictability of BG levels is a key issue since the glucose-insulin dynamics presents great inter- and intra-patient variability. Here, the question arises as to whether glycemic variability is not properly taken into account in models formulations and whether or it would compromise proper estimation of plasma insulin concentration. In order to tackle this point, a deterministic model describing glucose-insulin interaction plus a stochastic process to account for BG fluctuations were incorporated into the extended (EKF), cubature (CKF) and unscented (UKF) configurations of the Kalman filter to provide an estimate of the plasma insulin concentration. We found that for low glycemic variability, insulin state estimation can be attained with acceptable accuracy; however, as glycemic variability rises, Kalman filters rapidly degrade their performance as a consequence of large nonlinearities. Fil: Avila, Luis Omar. Universidad Nacional de San Luis. Facultad de Ciencias Físico Matemáticas y Naturales. Departamento de Informática. Laboratorio Investigación y Desarrollo en Inteligencia Computacional; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: de Paula, Mariano. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigaciones en Física e Ingeniería del Centro de la Provincia de Buenos Aires. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigaciones en Física e Ingeniería del Centro de la Provincia de Buenos Aires. - Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Centro de Investigaciones en Física e Ingeniería del Centro de la Provincia de Buenos Aires; Argentina Fil: Sánchez Reinoso, Carlos Roberto. Universidad Nacional de Catamarca. Facultad Tecnología y Ciencias Aplicadas. Centro de Investigación y Desarrollo en Modelado, Simulación y Optimización de Sistemas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

The determination of ethanol intoxication in whole blood samples may open the opportunity for a precise and quick point-of-measurement in the ambit of medical emergency or law enforcement. In contrast with traditional techniques based on breath sampling, direct blood measurements present greater immunity to errors specially in case of unconscious or non-collaborative patients. In this context, a portable, sensitive and easy-to-use instrument is highly desirable. In the current work we present a smartphone-based µPotentiostat which combines a novel circuital technique for sensor readout digitalization with a reusable lab-on-a-chip (LoC) concept. Such system allows both chronoamperometric and cyclic voltammetry measurements with a reduced number of electronic components on a very compact PCB (38.5 × 22.5 mm2). Power, data-link and user interface are provided in combination with a standard smartphone, enabling cost-effectiveness and reconfigurability without sacrificing precision. The readout platform discussed in this work has been coupled to a LoC for point-of-care combining Pt electrodes microfabricated on silicon substrate for electrochemical measurement and a microfluidic structure of methacrylate for fluid management. Biosensing is enabled by in situ electrodeposition of a calcium alginate hydrogel containing horseradish peroxidase (HPR) and alcohol oxidase (AOx) for selective ethanol detection. Alginate membrane electrodeposition has been here optimized for rapid generation (2 min) and to retain the cellular fraction, thus allowing the measurement in whole blood samples. The µPotentiostat features a sensitivity of 36 nA/g L−1 to ethanol concentration in blood in the 0–1.25 g;L−1 range, with a limit of quantification (LoQ) of 4.5 nA, which is a suitable response for discerning the legal, illegal, severely illegal thresholds in a 40 µL sample of blood.

Acenocoumarol (Sintrom®) is an oral anticoagulant prescribed for the treatment of a variety of thromboembolic disorders such as atrial fibrillation and thrombosis or embolism. It inhibits fibrin production preventing clot formation. Acenocoumarol has a narrow therapeutic range, and its effects depend on several factors, such as body weight, age, metabolism, diet, certain medical conditions or the intake of additional drugs, among others. A higher dose may result in the risk of bleeding, while if it is too low, the risk of blood clot can increase. Complementary tools that allow the therapeutic drug monitoring (TDM) of acenocoumarol plasmatic levels from the starting of the treatment would be of paramount importance to personalize the treatment. Point-of-care (POC) devices can offer an added value in facilitating on-site monitoring (i.e. hospitals, primary care doctor or even by the patient itself) and can aid in dosage management. With this aim, we have developed a compact and simple nanoplasmonic sensing device based on gold nanodisks for the rapid monitoring of acenocoumarol, using highly specific polyclonal antibodies produced against this drug. A specific and reproducible label free indirect competitive assay has been developed and the viability of performing the evaluation directly in plasma diluted 1:1 has been demonstrated. A limit of detection (LOD) of only 0.77 ± 0.69 nM, an IC50 of 48.2 ± 5.12 nM and a dynamic range between 3.38 ± 1.33 nM and 1154 ± 437 nM were achieved, which easily fit within the drug plasma levels of acenocoumarol, making this approach a highly attractive option for its decentralized monitoring in human plasma., The ICN2 is funded by the CERCA programme/Generalitat de Catalunya. The ICN2 is supported by the Severo Ochoa programme of the Spanish Ministry of Economy, Industry and Competitiveness (MINECO, grant no. SEV-2013-0295). The Nb4D group (formerly Applied Molecular Receptors group, AMRg) is a consolidated research group (Grup de Recerca) of the Generalitat de Catalunya and has support from the Department d’Universitats, Recerca i Societat de la Informació de la Generalitat de Catalunya (2017 SGR 1441). CIBER-BBN is an initiative funded by the Spanish National Plan for Scientific and Technical Research and Innovation 2013–2016, Iniciativa Ingenio 2010, Consolider Program, CIBER Actions are financed by the Instituto de Salud Carlos III with assistance from the European Regional Development Fund.

Tocopherols and tocotrienols are metabolized via hydroxylation and oxidation of their hydrophobic side chain to generate 13′-hydroxychromanols (13′-OHs) and various carboxychromanols, which can be further metabolized by conjugation including sulfation. Recent studies indicate that long-chain carboxychromanols, especially 13′-carboxychromanol (13′-COOH), appear to be more bioactive than tocopherols in anti-inflammatory and anticancer actions. To understand the potential contribution of metabolites to vitamin E-mediated effects, an accurate assay is needed to evaluate bioavailability of these metabolites. Here we describe an LC/MS/MS assay for quantifying vitamin E metabolites using negative polarity ESI. This assay includes a reliable sample extraction procedure with efficacy of ≥ 89% and interday/intraday variation of 3–11% for major metabolites. To ensure accurate quantification, short-chain, long-chain, and sulfated carboxychromanols are included as external/internal standards. Using this assay, we observed that sulfated carboxychromanols are the primary metabolites in the plasma of rodents fed with γ-tocopherol or δ-tocopherol. Although plasma levels of 13′-COOHs and 13′-OHs are low, high concentrations of these compounds are found in feces. Our study demonstrates an LC/MS/MS assay for quantitation of sulfated and unconjugated vitamin E metabolites, and this assay will be useful for evaluating the role of these metabolites in vivo.

The aim of this study was to evaluate the effects of geese’s maternal diet supplemented with flaxseed on the fatty acid profiles of egg yolks and the antioxidant status of their offspring. A total of 288 female Huoyan geese (42 weeks old) were randomly allotted to four experimental groups in this 56-day experiment and fed on diets containing flaxseed at 0% (control), 5%, 10% and 15%, respectively. There were nine replicate pens per treatment, with eight geese per replicate pen. The concentration of α-linolenic acid (linear, P

This study was conducted by two trials to investigate effects of sequential feeding with low- and high-protein diets on growth traits and plasma metabolites in geese. In Trial I, the effect of sequential feeding under time-restricted feeding system was investigated. Seventy-two White Roman goslings were randomly allotted into either sequential feeding (S1) or control feeding (C1) group. All goslings were fed for 1 h at morning and at evening, respectively, from 2 to 8 weeks of age. S1 group was offered 13% CP diet at morning and 19% CP diet at evening. C1 group was offered the same diet (16% CP; mixed equally with the two diets mentioned above) at both morning and evening. Blood samples were hourly collected for 4 h after feeding at both morning and evening for the determination of the postprandial plasma levels of glucose, triacylglycerol and uric acid at the end of experiment. Results showed that BW, average daily gain (ADG), and daily feed intake (FI) were not different between groups, but the feed efficiency (FE) in S1 group was significantly higher than that in C1 group (P

International audience; Based on the potential benefits for long-term human health, there is interest in developing sustainable nutritional strategies for lowering medium-chain saturated fatty acids (FA) and increasing specific unsaturated FA in ruminant milk. Dietary supplements of extruded linseeds (EL), fish oil (FO) or a mixture of EL and FO increase cis-9,trans-11 CLA and long-chain n-3 polyunsaturated FA in bovine milk. Supplements of FO cause milk fat depression in lactating cows, but information for dairy goats is limited. A total of 14 Alpine goats were used in a replicated 3×3 Latin square with 28-days experimental periods to examine the effects of EL alone or in combination with FO on animal performance, milk fat synthesis and milk FA composition. Treatments comprised diets based on natural grassland hay supplemented with no additional oil (control), 530 of EL or 340 g/day of EL and 39 g/day of FO (ELFO). Compared with the control, ELFO tended (P=0.08) to lower milk fat yield, whereas EL increased (P

An amperometric sensor for the determination of epinephrine (EP) was fabricated by modifying the carbon paste electrode (CPE) with pristine multi-walled carbon nanotubes (pMWCNTs) using bulk modification followed by drop casting of sodium dodecyl sulfate (SDS) onto the surface for its optimal potential application. The modified electrode showed an excellent electrocatalytic activity towards EP by decreasing the overpotential and greatly enhancing the current sensitivity. FE-SEM images confirmed the dispersion of pMWCNTs in the CPE matrix. EDX analysis ensured the surface coverage of SDS. A comparative study of pMWCNTs with those of oxidized MWCNTs (MWCNTsOX) modified electrodes reveals that the former is the best base material for the construction of the sensor with advantages of lower oxidation overpotential and the least background current. The performance of the modified electrode was impressive in terms of the least charge transfer resistance (Rct), highest values for diffusion coefficient (DEP) and standard heterogeneous electron transfer rate constant (k°). Analytical characterization of the modified electrode exhibited two linear dynamic ranges from 1.0×10-7 to 1.0×10-6 M and 1.0×10-6 to 1.0×10-4 M with a detection limit of (4.5±0.18)×10-8 M. A 100-fold excess of serotonin, acetaminophen, folic acid, uric acid, tryptophan, tyrosine and cysteine, 10-fold excess of ascorbic acid and twofold excess of dopamine do not interfere in the quantification of EP at this electrode. The analytical applications of the modified electrode were demonstrated by determining EP in spiked blood serum and adrenaline tartrate injection. The modified electrode involves a simple fabrication procedure, minimum usage of the modifier, quick response, excellent stability, reproducibility and anti-fouling effects.

Dogs play an important role in infectious disease transmission as reservoir hosts of many zoonotic and wildlife pathogens. Nevertheless, unlike wildlife species involved in the life cycle of pathogens, whose health status might be a direct reflection of their fitness and competitive abilities, dog health condition could be sensitive to socio-economic factors impacting the well-being of their owners. Here, we compare several dog health indicators in three rural communities of Panama with different degrees of socio-economic deprivation. From a total of 78 individuals, we collected blood and fecal samples, and assessed their body condition. With the blood samples, we performed routine hematologic evaluation (complete blood counts) and measured cytokine levels (Interferon-γ and Interleukin-10) through enzyme-linked immunosorbent assays. With the fecal samples we diagnosed helminthiases. Dogs were also serologically tested for exposure to Trypanosoma cruzi and canine distemper virus, and molecular tests were done to assess T. cruzi infection status. We found significant differences between dog health measurements, pathogen prevalence, parasite richness, and economic status of the human communities where the dogs lived. We found dogs that were less healthy, more likely to be infected with zoonotic pathogens, and more likely to be seropositive to canine distemper virus in the communities with lower economic status. This study concludes that isolated communities of lower economic status in Panama may have less healthy dogs that could become major reservoirs in the transmission of diseases to humans and sympatric wildlife., Acta Tropica, 135, pp.67-74; 2014

This study assessed the effects of dietary supplementation with extruded linseed on milk yield and composition, milk fatty acid (FA) profile and renal and hepatic metabolism of grazing goats in mid-lactation. Forty Saanen goats were divided into two isoproductive groups: one group was fed the control diet (CON) composed of hay and pelleted concentrate and the other group was supplemented with additional 180 g/day of extruded linseed (LIN; dry matter basis), which supplied 70 g/day of fat per head for 9 weeks. Animals grazed on pasture for ∼3 h/day after the first of the 2 daily milkings. Milk samples were collected weekly and analyzed for fat, protein, lactose, milk urea nitrogen (MUN) and somatic cell count. Blood samples were collected every 2 weeks and analyzed for total bilirubin, creatinine, aspartate transaminase (AST), alanine transaminase (ALT), gamma glutamyl transpeptidase, alkaline phosphatase, total protein and urea nitrogen. Milk yield was higher in the LIN than in the CON group (2369 v. 2052 g/day). LIN group had higher milk fat (37.7 v. 33.4 g/kg) and protein (30.7 v. 29.1 g/kg) concentration and lower MUN (35.0 v. 43.3 mg/dl) than CON group. Goats fed LIN had greater proportions of 18:1 trans11, 18:2 cis9trans11 and total polyunsatured fatty acids n-3 in milk fat, because of higher 18:3n-3 and 20:5n-3 FA, and lower proportions of short- and medium-chain FAs than goats fed CON. All kidney and liver function biomarkers in serum did not differ between dietary groups, except for AST and ALT, which tended to differ. Extruded linseed supplementation to grazing mid-lactating goats for 2 months can enhance the milk performance and nutritional profile of milk lipids, without altering the general hepatic and renal metabolism.

Chronic kidney disease (CKD) patients exhibit high levels of genetic damage. Part of this genetic damage is supposed to be caused by the hemodialysis (HD) therapy. Different and more efficient HD procedures could reduce the genetic damage and improve health status of CKD patients. In the present study, we analyzed if changing to online hemodiafiltration (OL-HDF) has a beneficial effect on the levels of genetic damage. The levels of genetic damage (DNA breaks and oxidatively damaged DNA) were analyzed in peripheral blood lymphocytes by using the comet assay. Forty-nine patients submitted to HD, 34 of them changing to OL-HDF and 15 patients continuing in low-flux HD, were included in the study. Plasma antioxidant capacity was also determined. Second sampling period was established after 6 months on the new or traditional HD protocol. A slight decrease in the levels of DNA damage was observed in patients who switched to OL-HDF (P=0.048) in relation to the reference group. This reduction is indicative that OL-HDF shows greater efficiency than low-flux HD in the reduction of basal levels of genetic damage. (C) 2016 Elsevier B.V. All rights reserved.

Hydroxytyrosol (HT) from olive oil, a potent bioactive molecule with health benefits, has a poor bioavailability, its free form (free HT) being undetectable so far. This fact leads to the controversy whether attained HT concentrations after olive oil polyphenol ingestion are too low to explain the observed biological activities. Due to this, an analytical methodology to determine free HT in plasma is crucial for understanding HT biological activity. Plasma HT instability and low concentrations have been major limitations for its quantification in clinical studies. Here, we describe a method to detect and quantify free HT in human plasma by using liquid chromatography coupled to tandem mass spectrometry. The method encompasses different steps of sample preparation including plasma stabilization, protein precipitation, selective derivatization with benzylamine, and purification by solid-phase extraction. A high sensitivity (LOD, 0.3ng/mL), specificity and stability of HT is achieved following these procedures. The method was validated and its applicability was demonstrated by analyzing human plasma samples after olive oil intake. A pharmacokinetic comparison was performed measuring free HT plasma concentrations following the intake of 25mL of ordinary olive oil (nearly undetectable concentrations) versus an extra-virgin olive oil (Cmax=4.40ng/mL). To our knowledge, this is the first time that an analytical procedure for quantifying free HT in plasma after olive oil dietary doses has been reported. The present methodology opens the door to a better understanding of the relationship between HT plasma concentrations and its beneficial health effects. This work was supported by grants from Instituto de Salud Carlos III FEDER, (PI14/00072), the CICYT-FEDER AGL2009-13517- C03-01 and AGL2012-40144-C03-01), grants from DIUE of the Generalitat de Catalunya (2014 SGR 680). CIBER de Fisiopatología de la Obesidad y Nutrición (CIBEROBN) is an initiative of the Instituto de Salud Carlos III, Madrid, Spain. J.R-M. was supported by a FI-DGR2012 predoctoral fellowship from the Generalitat de Catalunya. C.P-M. was supported by a Juan Rodés contract (JR15/00005).

A high degree of uremia is common in patients with end-stage renal disease and has been linked to the development of chronic inflammation and cardiovascular diseases. In conditions where transplantation is not possible, uremia can be reduced by hemodialysis although the repeated interventions have been implicated in loss of renal function, partially as a result of chronic inflammation and/or oxidative stress processes. In this context, it has been suggested that myeloperoxidase (MPO) can contribute to the oxidative stress during hemodialysis and to the cardiovascular risk. Protein damages due to MPO activity have never been assessed during hemodialysis although two of its reaction products, 3-chlorotyrosine and homocitrulline, are of interest. Indeed, the first one is a specific product of MPO activity and the formation of the second one could be catalyzed by MPO. In order to analyze these products in plasma proteins, a total hydrolysis method followed by liquid chromatography mass spectrometry analysis was developed. Different conditions of hydrolysis were tested and the optimized procedure was assessed for complete hydrolysis and artifactual chlorination. Finally, the method was used for analyzing 3-chlorotyrosine and homocitrulline in plasma proteins during a hemodialysis session in fifteen patients and data were related to measurements of MPO concentration and activity. Both increases in MPO activity and protein-bound 3-chlorotyrosine were observed, highlighting the involvement of MPO in oxidative stress during hemodialysis and further demonstrating the link between hemodialysis and cardiovascular diseases.

Lipid extraction from biological samples is a critical and often tedious preanalytical step in lipid research. Primarily on the basis of automation criteria, we have developed the BUME method, a novel chloroform-free total lipid extraction method for blood plasma compatible with standard 96-well robots. In only 60 min, 96 samples can be automatically extracted with lipid profiles of commonly analyzed lipid classes almost identically and with absolute recoveries similar or better to what is obtained using the chloroform-based reference method. Lipid recoveries were linear from 10–100 µl plasma for all investigated lipids using the developed extraction protocol. The BUME protocol includes an initial one-phase extraction of plasma into 300 µl butanol:methanol (BUME) mixture (3:1) followed by two-phase extraction into 300 µl heptane:ethyl acetate (3:1) using 300 µl 1% acetic acid as buffer. The lipids investigated included the most abundant plasma lipid classes (e.g., cholesterol ester, free cholesterol, triacylglycerol, phosphatidylcholine, and sphingomyelin) as well as less abundant but biologically important lipid classes, including ceramide, diacylglycerol, and lyso-phospholipids. This novel method has been successfully implemented in our laboratory and is now used daily. We conclude that the fully automated, high-throughput BUME method can replace chloroform-based methods, saving both human and environmental resources.

Most athletic horses are fed a high-starch diet despite the risk of health problems. Replacing starch concentrate with high-energy forage would alleviate these health problems, but could result in a shift in major substrates for muscle energy supply from glucose to short-chain fatty acids (SCFA) due to more hindgut fermentation of fibre. Dietary fat inclusion has previously been shown to promote aerobic energy supply during exercise, but the contribution of SCFA to exercise metabolism has received little attention. This study compared metabolic response with exercise and lactate threshold (VLa4) in horses fed a forage-only diet (F) and a more traditional high-starch, low-energy forage diet (forage-concentrate diet - FC). The hypothesis was that diet F would increase plasma acetate concentration and increase VLa4 compared with diet FC. Six Standardbred geldings in race training were used in a 29-day change-over experiment. Plasma acetate, non-esterified fatty acids (NEFA), lactate, glucose and insulin concentrations and venous pH were measured in samples collected before, during and after a treadmill exercise test (ET, day 25) and muscle glycogen concentrations before and after ET. Plasma acetate concentration was higher before and after exercise in horses on diet F compared with diet FC, and there was a tendency (P = 0.09) for increased VLa4 on diet F. Venous pH and plasma glucose concentrations during exercise were higher in horses on diet F than diet FC, as was plasma NEFA on the day after ET. Plasma insulin and muscle glycogen concentrations were lower for diet F, but glycogen utilisation was similar for the two diets. The results show that a high-energy, forage-only diet alters the metabolic response to exercise and, with the exception of lowered glycogen stores, appears to have positive rather than negative effects on performance traits.

The quality of the laboratory diagnostic approach in farm animals can be severely affected by pre-analytical factors of variation. They induce increase/decrease of biochemical and hematological analyte concentrations and, as a consequence, they may cause unsuitable conclusions and decisions for animal health management and research projects. The pre-analytical period covers the preparation of sampling, the sampling procedure itself, as well as all specimen handling until the beginning of the specific laboratory analysis. Pre-analytical factors may have either an animal-related or a technique-related background. Animal-related factors cover daytime/season, meals/fasting, age, gender, altitude, drugs/anesthesia, physical exercise/stress or coinfection. Technique-related factors are the choice of the tube including serum v. plasma, effects of anticoagulants/gel separators, the anticoagulant/blood ratio, the blood collection procedure itself, specimen handling, contamination, labeling, storage and serum/plasma separation, transportation of the specimen, as well as sample preparation before analysis in the laboratory. It is essential to have proper knowledge about the importance and source of pre-analytical factors to alter the entire diagnostic process. Utmost efforts should be made to minimize controllable factors. Analytical results have to be evaluated with care considering that pre-analytical factors of variation are possible causes of misinterpretation.

Cooling to 33.5 °C in babies with neonatal encephalopathy significantly reduces death and disability, however additional therapies are needed to maximize brain protection. Following hypoxia–ischemia we assessed whether inhaled 45–50% Argon from 2–26 h augmented hypothermia neuroprotection in a neonatal piglet model, using MRS and aEEG, which predict outcome in babies with neonatal encephalopathy, and immunohistochemistry. Following cerebral hypoxia–ischemia, 20 Newborn male Large White piglets, Graphical abstract, Highlights • 45–50% Argon augments brain protection above hypothermia after neonatal hypoxia–ischemia. • No physiological or biochemical change occurred with the 24 h exposure of 45–50% Argon. • 45–50% Argon led to faster recovery of aEEG, and improved cerebral metabolism on MRS. • TUNEL + cells were ~ 24 points lower per mm2 with Argon augmented cooling v cooling. • Argon was practical to administer with a standard neonatal ventilator (unlike xenon).

The composition of lipoproteins and the association of proteins with various particles are of much interest in the context of cardiovascular disease. Here, we describe a technique for the multidimensional analysis of lipoproteins and their associated apolipoproteins. Plasma is separated by size exclusion chromatography (SEC), and fractions are analyzed by reverse-phase arrays. SEC fractions are spotted on nitrocellulose slides and incubated with different antibodies against individual apolipoproteins or antibodies against various apolipoproteins. In this way, tens of analytes can be measured simultaneously in 100 μl of plasma from a single SEC separation. This methodology is particularly suited to simultaneous analysis of multiple proteins that may change their distribution to lipoproteins or alter their conformation, depending on factors that influence circulating lipoprotein size or composition. We observed changes in the distribution of exchangeable apolipoproteins following addition of recombinant apolipoproteins or interaction with exogenous compounds. While the cholesteryl ester transfer protein (CETP)-dependent formation of pre-β-HDL was inhibited by the CETP inhibitors torcetrapib and anacetrapib, it was not reduced by the CETP modulator dalcetrapib. This finding was elucidated using this technique.

Conserving irreplaceable, archived serum samples may sometimes conflict with the objective of minimizing measurement error due to laboratory effects. We sought to determine whether we could successfully combine assay results for DDT-related compounds and polychlorinated biphenyls (PCBs) in serum from the same birth cohort obtained from different laboratories over time. Using the Child Health and Development Studies (CHDS) serum archive, we compared variability for assays of a quality control pool to variability for assays of subject serum. The quality control pool was created from native archived serum samples that were pooled, then aliquoted, blinded and inserted pair-wise into assay batches along with the subject serum for 5 studies using CHDS samples conducted over a 13 year period by three different laboratories. We found that the variability between laboratory and over time within laboratory was small relative to inter-individual variability for p,p′-DDT (1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane), p,p′-DDE (1,1′-dichloro-2,2′-bis(p-chlorophenyl)ethylene) and o,p′-DDT (1,1,1-trichloro-2-(p-chlorophenyl)-2-(o-chlorophenyl)-ethane). Results were also consistent for most PCB congeners which were detectable in 85% or more of samples. Our results suggest that it is possible to combine assays for DDT and PCB congeners measured at positive levels as they are accumulated for cohort subjects without risking meaningful misclassification due to variation stemming from laboratory or time period. This has significant implications for future study costs, conservation of irreplaceable archived samples and for leveraging past investments for future research. For PCB congeners with very low levels, findings caution against pooling of assays without further exploration. Keywords: CV, Assay library, Organochlorines, Measurement error, Biomarkers, Epidemiology

Mice were gavaged with either 14C-labeled 2,2′5,5′ tetrachlorobiphenyl; 3,3′,4,4′ tetrachlorobiphenyl; or perfluorooctanoic acid. Absorption of these compounds was determined by assay of feces collected for 48 h after the gavage. Part of the animals received test diets containing olestra during this 48-hour period to determine its effect on absorption of the compounds. Mice that received the diet without olestra during this period were divided into groups that either continued the diet without olestra or changed to a diet containing olestra. These diets were continued for 7 days, and a second 48-hour fecal collection was made to measure the effect of olestra on enterohepatic circulation of the compounds and their metabolites. The animals were sacrificed, and blood, fat, and liver concentrations of 14C were measured. Olestra decreased the absorption of 2,2′,5,5′ tetrachlorobiphenyl. It also reduced tissue and blood concentrations of this compound. Olestra also decreased the absorption of 3,3′,4,4′ tetrachlorobiphenyl, but it did not alter enterohepatic circulation or tissue concentrations. Olestra significantly increased the excretion of perfluorooctanoic acid in the second 48-hour collection, suggesting an effect on enterohepatic circulation. It did not, however, alter tissue concentrations of perfluorooctanoic acid. These data are consistent with previously observed effects of olestra on the absorption and storage of lipophilic compounds. Keywords: Olestra, Polychlorinated biphenyl, PCB, Perfluorooctanoic acid, Rat, Absorption