S-methylcysteine sulphoxide,the brassica anaemia factor (a valuable dietary factor for man?)

Ruminant animals fed mainly or exclusively on brassica crops may develop a severe Heinz body anaemia. Clinical signs of the disease include haemoglobinuria, jaundice, increase in pulse rate and fall in milk production; there may also be growth retardation. Nitrate and thiocyanates were excluded as primary haemolytic factors in kale which contains up to 2% in DM of S-methylcysteine sulphoxide (SMCO). In test goats, daily intakes of SMCO in the range 15 to 19 g/100 kg liveweight, whether given as kale or the pure synthetic compound, elicited comparable haemolytic responses. SMCO was thus implicated as the major primary toxin in brassica poisoning. Rumen organisms converted SMCO into dimethyl disulphide which mimicked the haemolytic action of kale and SMCO in goats and is thus considered to be the secondary haemolysin. The practical consequence of this disulphide mechanism is that, because there is no simple way of counteracting the systemic action of dimethyl disulphide, prophylactic measures are restricted to limiting the intake of SMCO or to inhibiting disulphide production. Manipulation of rumen flora by dietary means to exclude organisms with SMCO-lyase is seen as a possible way of minimising the problem which in the long term however calls for developing brassica varieties with low SMCO content. The SMCO content of leafy brassicas increases as the plants mature; within a single plant highest SMCO levels are found in young leaves and growing shoots. SMCO is a heritable character and its selection will probably not lead to marked changes in protein or carbohydrate contents. In steers acute haemolytic response with growth inhibition was obtained with daily SMCO intakes, given as cabbage, of 18 to 35 g/100 kg liveweight. At the lower daily intake of 10–15 g the response was sub-acute, but the low grade anaemia produced would probably be incompatible with maximum productivity. Daily intakes below 10 g/100 kg generally lead to mild disturbances only in blood picture; and it is speculated that, on balance, at this level the effects of SMCO may even be beneficial to the animal. The possible prophylactic action of the S-alkylcysteine sulphoxides in coronary heart disease in man is briefly discussed.Resume Les ruminants nourris principalement ou exclusivement de crucifères peuvent contracter une grave anémie à corpuscules endoglobulaires. Les signes cliniques de la maladie comprennent l'hémoglobinurie, la jaunisse, l'élévation du pouls et la baisse de la production de lait et éventuellement aussi un retard de la croissance. Le nitrate et les thiocyanates ont été exclus comme facteurs hémolytiques primaires du chou à feuilles, qui contient jus qu'à 2% en matières sèches du sulfoxyde de s-méthylcystéine (SMCO). Chez les chèvres utilisées comme sujets d'expériences, des doses quotidiennes de SMCO de l'ordre de 15 à 19 g par 100 kg de poids corporel, absorbée sous la forme de chou ou sous celle de composé synthétique pur ont donné des réactions hémolytiques comparables. On a donc conclu que le SMCO était la principale toxine primaire responsable de l'empoisonnement brassicaire. Les organismes du rumen ont transformé le SMCO en disulphide de diméthyle qui a reproduit l'action hémolytique du chou et du SMCO chez les chèvres, et qui est donc considéré comme l'hémolysine secondaire. Étant donné qu'il n'y a pas de moyens simples de contrecarrer l'action systémique du disulphide de diméthyle, les seules mesures prophylactiques sont la limitation de l'absorption de SMCO ou l'inhibition de la production de disulphide. L'action sur la flore du rumen par des moyens diététiques, à fin d'exclure les organismes à SMCO-lyase, semble un moyen possible d'atténuer le problème. Mais à long terme, il faudra développer des variétés de crucifères à faible teneur en SMCO. La teneur en SMCO des crucifères feuillus augmente au fur et à mesure de la maturation. Sur une même plante, les teneurs les plus élevées se rencontrent dans les jeunes feuilles et dans les pousses en croissance. Le SMCO est un caractère héréditaire et une sélection operée en fonction de lui n'entraînera probablement pas de modifications importantes des teneurs en protéines ou en hydrates de carbone. Une réaction hémolytique aiguë avec inhibition de la croissance a été provoquée chez les bouvillons au moyen de doses quotidiennes de SMCO de 18 à 35 g par 100 kg de poids corporel, administrées sous forme de chou. A la dose quotidienne plus faible de 10 à 15 g la réaction a été sub-aiguë mais l'anémie faible qui en résulte serait probablement incompatible avec une productivité optimale. Les doses quotidiennes inférieures à 10 g par 100 kg ne provoquent généralement que de légères perturbations de la formule hématologique. On estime qu'à ce taux les effets du SMCO peuvent même, dans l'ensemble, être bénéfiques à l'animal. L'action prophylactique possible des sulfoxydes de s-alkylcysteine dans les cas de maladies coronaires chez l'homme est examinée brièvement.

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Kurzfassung Bei Wiederkäuern, die hauptsächlich oder ausschliesslich Brassica-Futter erhalten, kann es zu einer schweren Innenkörperanämie kommen. Klinische Anzeichen hierfür sind u.a. Bluthernen, Gelbsucht, erhöhte Pulsfrequenz und Rückgang der Milcherzeugung, ausserdem unter Umständen ein verlangsamtes Wachstum. Nitrat und Thiocyanat wurden als primäre hämolytische Faktoren im Brassica oleracea, das bis zu 2% S-Methylcystein sulfoxid (SMCO) in der Trockenmasse enthält, ausgeschlossen. Bei Test-Ziegen führten Tagesdosen von SMCO im Bereich von 15 bis 19 g/100 kg Lebendgewicht in Form von Brassica oleracea oder dem reinen, synthetisierten Bestandteil zu vergleichbaren hämolytischen Reaktionen. SMCO war also als wichtigstes Primärtoxin bei Brassica-Vergiftungen beteiligt. Die Pansenorganismen setzten SMCO in Dimethyldisulfid um, das die selbe hämolytische Wirkung wie Brassica oleracea und SMCO bei Ziegen hatte und so als sekundäres Hämolysin gilt. Die praktische Folge dieses Disulfidmechanismus ist, dass vorbeugende Massnahmen auf die Begrenzung der SMCO-Aufnahme oder die Verhinderung der Disulfidproduktion beschränkt sind, da es keine einfache Möglichkeit gibt, der systematischen Aktion von Dimethyldisulfid entgegenzuwirken. Eine Manipulation der Pansenflora durch entsprechende Ernährung zur Ausschaltung von Organismen mit SMCO-Lyase gilt als Möglichkeit zur Minimalisierung des Problems, das langfristig gesehen jedoch die Entwicklung von Brassica-Arten mit geringem SMCO-Gehalt erforderlich macht. Der SMCO-Gehalt von Blattkohl nimmt mit zunehmender Reife der Pflanze zu; innerhalb einer Pflanze findet sich der grösste SMCO-Gehalt in jungen Blättern und wachsenden Trieben. SMCO ist ein erbliches Merkmal, und seine Selektion wird vermutlich nicht zu grösseren Aenderungen im Protein-oder Kohlehydratgehalt führen. Bei Ochsen wurde eine akute hämolytische Reaktion mit Wachstumsverzögerung bei täglichem SMCO-Dosen von 18 bis 35 g/100 kg Lebendgewicht in Form von Kohl erzielt. Bei der geringeren Tagesdosis von 10–15 g war die Reaktion subakut, die entwickelte geringfügige Anämie wäre jedoch vermutlich mit maximaler Produktivität unvereinbar. Tagesdosen unter 10 g/100 kg führen im allgemeinen nur im Blutbild zu geringen Störungen; man vermutet, dass alles in allem SMCO bei dieser Dosierung sogar eine positive Auswirkung auf das Tier haben kann. Die mögliche prophylaktische Wirkung der S-Alkylcystein sulfoxide bei Erkrankungen der Herzkranzgefässe beim Menschen wird kurz erörtert.

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Riassunto I ruminanti la cui dieta è in gran parte o esclusivamente rappresentata da cavoli (Brassica spp.) possono essere colpiti da una grave forma di anemia (Heinz body anaemia). I sintomi clinici della malattia comprendono emoglobinuria, ittero, aumento della frequenza cardiaca e caduta della produzione lattea. A volte si può anche avere ritardo della crescita. I nitrati e i tiocianati sono stati esclusi come fattori emolitici primari nei cavoli che contengono fino al 2% in DM di S-metilcisteina sulfossido (SMCO). In experimenti condotti sulle capre, la somministrazione giornaliera di SMCO in dosi di 15–19 g/100 kg di peso vivo, sia sotto forma di vegetale che di prodotto sintetico puro, ha provocato fenomeni emolitici relativamente simili. L'SMCO è stato pertanto indicato come la principale tossina che ha un ruolo primario nell'intossicazione da cavoli. La flora ruminale converte infatti l'SMCO in dimetil bisolfuro, il quale svolge un'azione emolitica simile a quella osservata nelle capre dopo l'assunzione di cavoli e di SMCO; il dimetil bisolfuro, pertanto, e ritenuto costituire l'emolisina secondaria. Le conseguenze pratiche di tale produzione di dimetil bisolfuro è che, non esistendo un metodo semplice per contrastare l'azione sistemica del dimetil bisolfuro, le misure profilattiche non possono essere volte ad altro se non a limitare l'assunzione di SMCO o ad inibire la produzione di bisolfuro. Gli interventi sulla flora del rumine mediante una dieta capace di eliminare i microrganismi provvisti di SMCO-liasi vengono considerati come un possibile mezzo per minimizzare il problema, il quale, tuttavia, se visto a lungo termine, richiede la creazione di varietà di cavoli a basso contenuto di SMCO. Il contenuto di SMCO delle foglie dei cavoli aumenta col maturare della pianta: nella pianta singola i livelli piu alti di SMCO si riscontrano nelle foglie giovani e nei germogli. La presenza di questa sostanza costituisce un carattere ereditario ed un intervento selettivo in tal senso non dovrebbe condurre a significative alterazioni del contenuto in proteine e carboidrati della pianta. Nei vitelloni e stato possibile indurre una risposta emolitica di tipo acuto somministrando giornalmente, sotto forma di cavoli, 18–35 g di SMCO per 100 kg di peso vivo. Un' assunzione giornaliera minore, pari a 10–15 g, ha dato luogo ad una forma emolitica subacuta, ma e tuttavia da ritenere che la lieve anemia cosi indotta non sia compatibile con una produttività ottimale. Livelli giornalieri al di sotto dei 10 g/100 kg hanno causato soltanto lievi alterazioni a carico del quadro ematico, ed è stata avanzata l'ipotesi che, tutto sommato, gli effetti dell'SMCO possano, a questi livelli, essere addirittura di beneficio all'animale. Viene discussa brevemente la possibile azione preventiva degli S-alchilcisteina sulfossidi sulle malattie cardio-coronariche dell'uomo.

     

Transforming growth factor β signaling in uterine development and function

Transforming growth factor β(TGFβ) superfamily is evolutionarily conserved and plays fundamental roles in cell growth and differentiation. Mounting evidence supports its important role in female reproduction and development. TGFBs1-3 are founding members of this growth factor family, however, the in vivo function of TGFβsignaling in the uterus remains poorly defined. By drawing on mouse and human studies as a main source, this review focuses on the recent progress on understanding TGFβ signaling in the uterus. The review also considers the involvement of dysregulated TGFβ signaling in pathological conditions that cause pregnancy loss and fertility problems in women.     

Expressions of tumor necrosis factor-α, its receptor I,II and receptor-associated factor 2 in the porcine corpus luteum during the estrous cycle and early pregnancy

We examined the gene and protein levels of tumor necrosis factor (TNF)-α, its receptors (types I and II, designated TNF-RI and TNF-RII, respectively), TNF receptor-associated factor 2 (TRAF2) and morphological features in the porcine corpus luteum (CL), on Days 13 and 17 (Day 0 = the last day of estrus) of the estrous cycle or of early pregnancy. Gene expression levels of TNF-α, TNF-RI, TNF-RII and TRAF2 were unaffected by the day or reproductive status. TNF-α concentration was significantly higher in the CL on Day 17 of pregnancy than on Day 13 of pregnancy and on day 17 of the estrous cycle. The TNF-RI protein level was significantly higher in the CL on Days 13 and 17 of pregnancy than those of the estrous cycle, significantly increasing on Day 17 compared with those on Day 13 in pregnancy. In relation to TNF-RII protein levels, although there were no change during pregnancy, there was a tendency (P?=?0.0524) to up-regulate as pregnancy proceeded. In estrous cycle, TNF-RII protein levels decreased significantly as luteolysis proceeded. TRAF2 protein level was significantly higher in the CL on Days 13 and 17 of pregnancy than during estrous. There were few apoptotic bodies in the CL between Days 13 and 17 of pregnancy than during esrous. There were few apoptotic bodies in the CL between Days 13 and 17 of pregnancy. The number of apoptotic bodies was much greater than the CL on Day 17 of the estrous than those of pregnancy. Thus, the TNF-α and TNF-RI and TNF-RII pathways including the TRAF2 protein, known to control of cell differentiation, tissue renewal and apoptosis, might participate in maintaining the porcine CL during early pregnancy.     

Relative condition factor and parasitism in anostomid fishes from the floodplain of the Upper Paraná River, Brazil

Individuals of four species of Leporinus were captured in the floodplain of the Upper Paraná River and their metazoan parasites were collected. Fifty-eight taxa of ectoparasites and endoparasites were recorded: 31 in Leporinus lacustris, 32 in Leporinus friderici, 28 in Leporinus obtusidens and 25 in Leporinus elongatus. The aim of this paper was to study the relationship between the host's condition and the parameters of infrapopulations and infracommunities. The health of the host was represented by the relative condition factor (Kn). Richness and number of individuals in the infracommunities of ectoparasites covariated with the Kn of the hosts in one species of fish. Some infrapopulations of ectoparasites covariated negatively and/or the mean Kn of parasitized individuals was lower than those without parasites. The abundance in some infrapopulations of endoparasites covariated positively with the Kn and/or the mean Kn of parasitized fish was better. These results may be related to different transmission strategies used by ecto and endoparasites.     

Fibroblast growth factor 21 in dairy cows:current knowledge and potential relevance

Fibroblast growth factor 21(FGF21)has been identified as an important regulator of carbohydrate and lipid metabolism,which plays an important role for metabolic regulation,particularly under conditions of energy deprivation or stress conditions.Dairy cows are subjected to a negative energy balance and various kinds of stress particularly during the periparturient phase and during early lactation.It has been shown that the plasma concentration of FGF21 in dairy cows is dramatically increased at parturition and remains high during the first weeks of lactation.This finding suggests that FGF21 might exert similar functions in dairy cows than in other species,such as mice or humans.However,the role of FGF21 in dairy cows has been less investigated so far.Following a brief summary of the previous findings about the function of FGF21 in humans and mice,the present review aims to present the current state of knowledge about the role of FGF21 in dairy cows.The first part of the review deals with the tissue localization of FGF21 and with conditions leading to an upregulation of FGF21 expression in the liver of dairy cows.In the second part,the influence of nutrition on FGF21 expression and the role of FGF21 for metabolic diseases in dairy cows is addressed.In the third part,findings of exogenous FGF21 application on metabolism in dairy cows are reported.Finally,the potential relevance of FGF21 in dairy cows is discussed.It is concluded that FGF21 might be of great importance for metabolic adaptation to negative energy balance and stress conditions in dairy cows.However,further studies are needed for a better understanding of the functions of FGF21 in dairy cows.     

Expression of matrix metalloproteinases in bovine luteal cells induced by prostaglandin F2α, interferon γ and tumor necrosis factor α

We recently demonstrated that luteal cells flow out from the ovary via lymphatic vessels during luteolysis. However, the regulatory mechanisms of the outflow of luteal cells are not known. Matrix metalloproteinases (MMPs) can degrade the extracellular matrix and basal membrane, and tissue inhibitors of matrix metalloproteinases (TIMPs) inhibit the activity of MMPs. To test the hypothesis that MMP expression in luteal cells is regulated by luteolytic factors, we investigated the effects of prostaglandin F2α (PGF), interferon γ (IFNG) and tumor necrosis factor α (TNF) on the mRNA expression of MMPs and TIMPs in cultured luteal cells. Luteal cells obtained from the CL at the mid-luteal stage (days 8–12 after ovulation) were cultured with PGF (0.01, 0.1, 1 μM), IFNG (0.05, 0.5, 5 nM) and TNF (0.05, 0.5, 0.5 nM) alone or in combination for 24 h. PGF and IFNG significantly increased the expression of MMP-1 mRNA. In addition, 1 μM PGF in combination with 5 nM IFNG stimulated MMP-1 and MMP-9 mRNA expression significantly more than either treatment alone. In contrast, IFNG significantly decreased the level of MMP-14 mRNA. The mRNA expression of TIMP-1, which preferentially inhibits MMP-1, was suppressed by 5 nM INFG. One μM PGF and 5 nM IFNG suppressed TIMP-2 mRNA expression. These results suggest a new role of MMPs: luteal MMPs stimulated by PGF and IFNG break down the extracellular matrix surrounding luteal cells, which accelerates detachment from the CL during luteolysis, providing an essential prerequisite for outflow of luteal cells from the CL to lymphatic vessels.     

Apoptosis and abundance of Bcl-2 family and transforming growth factor β1 signaling proteins in canine myxomatous mitral valves

ObjectivesTo determine the percentage of cells undergoing apoptosis within canine myxomatous valves and to evaluate whether TGFβ1 can be implicated as an anti-apoptosic signal through the Bcl-2 family of signaling proteins.AnimalsPost-mortem mitral valve leaflets harvested from 5 normal dogs, 5 dogs with early-stage myxomatous mitral valve disease (MMVD), and 5 dogs with late-stage MMVD.Materials and methodsThe number of cells expressing cleaved caspase-3, DNA fragmentation (TUNEL marker) and apoptotic bodies were evaluated as a measure of apoptosis. To evaluate the relationship between TGFβ1 signaling and apoptosis, the abundance of activated TGFβ1 signaling protein, phosphorylated Smad 2/3 (p-Smad 2/3), and Bcl-2 family proteins (pro-apoptotic Bax and anti-apoptotic Bcl-2) was determined by immunohistochemistry.ResultsCells in normal and both stages of MMVD expressed the TUNEL marker and cleaved caspase-3, but not apoptotic bodies. The percentage of TUNEL marker and cleaved caspase-3 positive nuclei was not significantly different between groups of dogs (p > 0.05). P-Smad 2/3 and Bax were more abundant in myxomatous mitral valves while Bcl-2 was less abundant. P-Smad 2/3 primarily increased in the atrialis layer and was abundantly increased only in late-stage MMVD.ConclusionsThese data suggest that interstitial cells in MMVD are in a pro-apoptotic condition; however, they do not execute apoptosis. Thus, apoptosis does not explain differences in cellular density in canine MMVD. TGFβ1 signaling through the canonical SMAD pathway is increased in myxomatous mitral valves, but does not apparently mediate interstitial cell apoptosis in canine MMVD.     

Luteolysis and associated interrelationships among circulating PGF2α, progesterone, LH, and estradiol in mares

The changing concentrations and temporal relationships among a PGF2α metabolite (PGFM), progesterone (P₄), LH, and estradiol-17β (E₂) before, during, and after luteolysis were studied in 10 mares. Blood samples were collected every hour for ≥4 d beginning on day 12 after ovulation. The luteolytic period extended from a decrease in P₄ at a common transitional hour (Hour 0) at the end of preluteolysis and beginning of luteolysis to a defined ending when P₄ reached 1 ng/mL. The length of luteolysis was 22.9 ± 0.9 h, contrasting with 2 d in published P₄ profiles from sampling every 6 to 24 h. In mares with complete data for Hours −40 to −2 (n = 6), PGFM concentrations remained below assay sensitivity (n = 2) or two or three small pulses (peak, 29 ± 4 pg/mL) occurred. During luteolysis, the pulses became more prominent (peak, 193 ± 36 pg/mL). Rhythmicity of PGFM pulses was not detected by a pulsatility program during preluteolysis but was detected in seven of nine mares during luteolysis and postluteolysis combined. The nadir-to-nadir interval for LH pulses and the peak-to-peak interval between adjacent pulses were longer (P < 0.05) during preluteolysis than during luteolysis (nadir to nadir, 5.2 ± 0.3 h vs 3.6 ± 0.4 h; peak to peak, 9.4 ± 1.0 h vs 4.7 ± 0.5 h). Unlike reported findings in cattle, concentrations of P₄ decreased linearly within the hours of each PGFM pulse during luteolysis, and a positive effect of an LH pulse on P₄ and E₂ concentration was not detected. The reported balancing of P₄ concentrations between a negative effect of PGF2α and a positive effect of LH in heifers was not detected in mares.     

Diagnostic orientation values for ACTH and other parameters for clinically healthy donkeys and mules (insulin,triglycerides, glucose,fructosamines, and ɣ-GT)

Pituitary pars intermedia dysfunction is the most prevalent endocrine disease in horses. Although donkeys and mules may also be affected, only a few data have been published. Reference values for diagnostic parameters, such as adrenocorticotropic hormone (ACTH), are especially scarce or even lacking. Therefore, in the present study, available data from the literature have been verified and completed to facilitate a reliable diagnosis. Clinical inspections and haematological and biochemical examinations were carried out four times in a three-month interval (February to November) in 44 donkeys and 31 mules. Data from clinically healthy animals were used as an orientation. Plasma ACTH concentrations showed seasonal changes in both animal groups. However, it was generally higher in donkeys than mules. Although blood glucose (EDTA plasma) showed no difference between groups, serum insulin concentrations were consistently higher in donkeys. Serum fructosamine levels were slightly higher in mules, whereas, in some cases, serum triglyceride levels were considerably higher in donkeys. Serum gamma-glutamyltransferase showed a striking peak in mules in August, whereas the remaining gamma-glutamyltransferase values were lower compared to donkeys. By comparing donkeys and mules, the present work reveals differences in various blood parameters which should be considered for diagnoses and future studies.     

Choosing among correlative,mechanistic, and hybrid models of species’ niche and distribution

Different models are available to estimate species’ niche and distribution. Mechanistic and correlative models have different underlying conceptual bases, thus generating different estimates of a species’ niche and geographic extent. Hybrid models, which combining correlative and mechanistic approaches, are considered a promising strategy; however, no synthesis in the literature assessed their applicability for terrestrial vertebrates to allow best-choice model considering their strengths and trade-offs. Here, we provide a systematic review of studies that compared or integrated correlative and mechanistic models to estimate species’ niche for terrestrial vertebrates under climate change. Our goal was to understand their conceptual, methodological, and performance differences, and the applicability of each approach. The studies we reviewed directly compared mechanistic and correlative predictions in terms of accuracy or estimated suitable area, however, without any quantitative analysis to support comparisons. Contrastingly, many studies suggest that instead of comparing approaches, mechanistic and correlative methods should be integrated (hybrid models). However, we stress that the best approach is highly context-dependent. Indeed, the quality and effectiveness of the prediction depends on the study's objective, methodological design, and which type of species’ niche and geographic distribution estimated are more appropriate to answer the study's issue.     

Hypoxia inducible factor 1α expression and effects of its inhibitors in canine lymphoma

Hypoxic conditions in various cancers are believed to relate with their malignancy, and hypoxia inducible factor-1α (HIF-1α) has been shown to be a major regulator of the response to low oxygen. In this study, we examined HIF-1α expression in canine lymphoma using cell lines and clinical samples and found that these cells expressed HIF-1α. Moreover, the HIF-1α inhibitors, echinomycin, YC-1 and 2-methoxyestradiol, suppressed the proliferation of canine lymphoma cell lines. In a xenograft model using NOD/scid mice, echinomycin treatment resulted in a dose-dependent regression of the tumor. Our results suggest that HIF-1α contributes to the proliferation and/or survival of canine lymphoma cells. Therefore, HIF-1α inhibitors may be potential agents to treat canine lymphoma.     

11-Hydroxy-β-steroid dehydrogenase gene expression in canine adipose tissue and adipocytes: stimulation by lipopolysaccharide and tumor necrosis factor α

The enzyme 11β-hydroxysteroid dehydrogenase 1 (11β-HSD-1) is expressed in a number of tissues in rodents and humans and is responsible for the reactivation of inert cortisone into cortisol. Its gene expression and activity are increased in white adipose tissue (WAT) from obese humans and may contribute to the adverse metabolic consequences of obesity and the metabolic syndrome. The extent to which 11β-HSD-1 contributes to adipose tissue function in dogs is unknown; the aim of the present study was to examine 11β-HSD-1 gene expression and its regulation by proinflammatory and anti-inflammatory agents in canine adipocytes. Real-time PCR was used to examine the expression of 11β-HSD-1 in canine adipose tissue and canine adipocytes differentiated in culture. The mRNA encoding 11β-HSD-1 was identified in all the major WAT depots in dogs and also in liver, kidney, and spleen. Quantification by real-time PCR showed that 11β-HSD-1 mRNA was least in perirenal and falciform depots and greatest in subcutaneous, omental, and gonadal depots. Greater expression was seen in the omental depot in female than in male dogs (P = 0.05). Gene expression for 11β-HSD-1 was also seen in adipocytes, from both subcutaneous and visceral depots, differentiated in culture; expression was evident throughout differentiation but was generally greatest in preadipocytes and during early differentiation, declining as cells progressed to maturity. The inflammatory mediators lipopolysaccharide and tumor necrosis factor α had a main stimulatory effect on 11β-HSD-1 gene expression in canine subcutaneous adipocytes, but IL-6 had no significant effect. Treatment with dexamethasone resulted in a significant time- and dose-dependent increase in 11β-HSD-1 gene expression, with greatest effects seen at 24 h (2nM: approximately 4-fold; 20nM: approximately 14-fold; P = 0.010 for both). When subcutaneous adipocytes were treated with the peroxisome proliferator activated receptor γ agonist rosiglitazone, similar dose- and time-dependent effects were noted. However, no effects were seen when adipocytes from the gonadal WAT depot were treated with rosiglitazone. The induction of 11β-HSD-1 expression, by the pro-inflammatory cytokine tumor necrosis factor α and by lipopolysaccharide may have implications for the pathogenesis of obesity and its associated diseases in the dog.     

Circadian and circannual rhythms of cortisol,ACTH, and α-melanocyte-stimulating hormone in healthy horses

Cosinor analysis was used to evaluate whether pituitary and adrenal hormones exhibit circadian rhythmicity in horses. The effect of season and animal age on their respective rhythms was also determined. In addition, the usefulness of evaluating cortisol rhythmicity for the diagnosis of pituitary pars intermedia dysfunction (PPID) was assessed. Serum cortisol concentrations (P < 0.01), but not plasma ACTH or α-melanocyte-stimulating hormone (α-MSH), showed a significant circadian periodicity in horses. An effect of season on hormone concentration was observed with plasma ACTH and α-MSH concentration greater in the fall and cortisol concentration greater in the spring (P < 0.001). Age did not affect cortisol rhythm, but it did blunt the variation in cortisol concentration in horses, similar to what has been previously reported to occur in aged people and dogs. In addition, our results suggest that clinically and diagnostically normal, non–PPID-affected horses commonly have a loss of cortisol diurnal rhythm. Therefore, measurement of circadian rhythm is not an appropriate diagnostic test for PPID.     

Activated platelet-derived growth factor β receptor and Ras-mitogen-activated protein kinase pathway in natural bovine urinary bladder carcinomas

Bovine papillomavirus types 1 or 2 (BPV-1/2) are involved in the aetiopathogenesis of bovine urinary bladder cancer. BPV-1/2 E5 activates the platelet-derived growth factor β receptor (PDGFβR). The aim of this study was to analyse the Ras/mitogen-activated protein kinase (MAPK) pathway in relation to activation of PDGFβR in natural bovine urinary bladder carcinomas. Co-immunoprecipitation and Western blot analysis demonstrated that recruitment of growth factor receptor bound protein 2 (GRB-2) and Sos-1 to the activated PDGFβR was increased in carcinomas compared to normal tissues. Higher grade bovine urinary bladder carcinomas were associated with activation of Ras, but not with activation of downstream mitogen-activated protein kinase/extracellular signal-regulated kinase (Mek 1/2) or extracellular signal-regulated kinase (Erk 1/2).     

Peripheral tumor necrosis factor α regulation of adipose tissue metabolism and adipokine gene expression in neonatal pigs

The neonatal pig is susceptible to stress and infection, conditions which favor tumor necrosis factor α (TNFα) secretion. This study examined whether TNFα can alter metabolic activity and cytokine gene expression within neonatal pig adipose tissue. Cell cultures were prepared from neonatal subcutaneous adipose tissue using standard procedures. Cultures (5 experiments) were incubated with medium containing ¹⁴C-glucose for 4 h to measure glucose conversion to lipid in the presence of combinations of TNFα (10 ng), insulin (10 nM) and an anti-pig TNFα antibody (5 μg). Basal lipogenesis was not affected by TNFα treatment (P?>?0.05). However, insulin stimulated lipogenesis was reduced by TNFα (P?<?0.02). For gene expression studies, cultures were incubated with 0, 2.5, 5.0 or 10 ng TNFα for 2, 4 or 24 h (n?=?4 experiments). Interleukin 6 and TNFα gene expression were acutely (2–4 h) stimulated by exogenous TNFα treatment (P?<?0.05), as analyzed by real-time PCR. Adiponectin mRNA abundance was reduced (P?<?0.001) while monocyte chemotactic gene expression was increased by TNFα treatment at all time points (P?<?0.001). Chronic treatment (24 h) was required to increase monocyte multiplication inhibitory factor or suppress lipoprotein lipase gene expression (P?<?0.02). These data suggest conditions which increase serum TNFα, like sepsis, could suppress lipid accumulation within adipose tissue at a time of critical need in the neonate and induce a variety of adipose derived cytokines which may function to alter adipose physiology.     

Effect of ovine granulocyte-macrophage colony-stimulating factor on bovine in vitro embryo development and blastocyst interferon-τ secretion

This experiment examined the effects of including recombinant ovine granulocyte-macrophage colony-stimulating factor (GMCSF) in in vitro culture on secretion of interferon-τ (IFNT) by bovine blastocysts. At 32 h post-insemination (p.i.), cleaved bovine zygotes were selected and incubated with or without GMCSF for either 48 h only (between 32 and 80 h p.i., Early) or until day 9 p.i. (Throughout). Concentrations of GMCSF (ng/ml) examined were as follows: Experiment 1: 2, 5, 10 and 50 (Early only); Experiment 2: 50 (Early and Throughout); Experiment 3: 2 and 10 (Early and Throughout). In none of the experiments did GMCSF have an effect (p > 0.05) on the numbers of blastocysts formed or blastocyst characteristics as assessed by cell number, proportion of apoptotic cells or oxidation of pyruvate. When GMCSF was included in culture medium between 32 and 80 h p.i. (Early), IFNT concentrations were lower (in media drops recovered after culture of groups of embryos for 48 h between days 7 and 9 p.i. and normalized by the numbers of blastocysts developing within each drop) compared to no inclusion of GMCSF or GMCSF present Throughout culture (Experiment 2, p > 0.05; Experiment 3, p = 0.038). IFNT was present in media drops in which groups of embryos had been incubated between days 7 and 9 p.i. but in which no blastocysts had developed. Experimental treatment did not influence (p > 0.05) IFNT secretion by blastocysts incubated individually for 24 h. However, during the 24-h individual culture, blastocysts recovered on day 7 secreted less IFNT than blastocysts recovered on day 8 (mean ± SE; 15 ± 1.3 v 30 ± 3.6 pg/ml; p < 0.001). In conclusion, in contrast to previous studies in the ovine, GMCSF did not increase IFNT secretion but in agreement with the ovine did not affect bovine blastocyst development.