Serum Lipid Profile in Iranian Fat‐tailed Sheep in Late Pregnancy,at Parturition and During the Post‐parturition Period

Blood samples were obtained from 12 Iranian fat‐tailed sheep during 7 weeks pre‐partum, at parturition and 7 weeks post‐partum. The lipids measured were cholesterol, triglyceride, total lipid, high‐density lipoprotein (HDL)‐cholesterol, low‐density lipoprotein (LDL)‐cholesterol, and very low‐density lipoprotein (VLDL)‐cholesterol. The concentrations of cholesterol, triglyceride, HDL‐cholesterol and VLDL‐cholesterol during the 7 weeks pre‐partum, at parturition and the 7 weeks post‐partum were significantly different (P < 0.05). One week before parturition, the concentrations of cholesterol, triglyceride, HDL‐cholesterol and VLDL‐cholesterol were higher (P < 0.05) than at other periods. The lowest concentrations of these parameters were observed 2–3 weeks after parturition. In this study, significant positive correlations were observed between the time of sampling (pre‐partum, parturition and post‐partum) and serum cholesterol (r=0.22; P < 0.01) and HDL‐cholesterol (r=0.25; P < 0.01).     

Large‐Scale Cross‐Sectional Serological Survey of Schmallenberg Virus in Belgian Cattle at the End of the First Vector Season

A cross‐sectional survey was conducted in the Belgian cattle population after the first period of infection of the emerging Schmallenberg virus. A total number of 11 635 cattle from 422 herds sampled between 2 January and 7 March 2012 were tested for the presence of Schmallenberg‐specific antibodies using an ELISA kit. Between‐herd seroprevalence in cattle was estimated at 99.76% (95% CI: 98.34–99.97) and within‐herd seroprevalence at 86.3% (95% CI: 84.75–87.71). An Intraclass Correlation Coefficient of 0.3 (P < 0.001) was found, indicating that the correlation between two animals within a herd with respect to their serological status was high. Those results corroborate the conclusion that the Schmallenberg virus was widespread in Belgium during winter 2011. Seroprevalence was shown to be statistically associated to the animal's age (P < 0.0001): with 64.9% (95% CI: 61.34–68.3) estimated for the 6–12 months of age, 86.79% (95% CI: 84.43–88.85) for the 12–24 months of age and 94.4% (95% CI: 93.14–95.44) for the animals older than 24 months. Based on the results of the described serological survey, we can conclude that after the first Schmallenberg virus episode, almost every Belgian cattle has already been in contact with the virus. In consequence, the vast majority of the host animals should have developed post infection protective immunity against the virus.     

Transferring and Distributing Profiles of p,p′‐(DDT) in Egg‐forming Tissues and Eggs of Laying Hens Following a Single Oral Administration

Laying hens were administered orally with a single dose of p,p′‐(DDT) (1 mg/kg bodyweight). The concentrations (μg/g) of DDT or its metabolites, p,p′‐(DDE) and p,p′‐(DDD), in the main tissues involved in egg formation (blood, liver, ovary, and oviducts) and egg yolk, collected 1 day after DDT dosing, were determined by normal‐phase high‐performance liquid chromatography. The limits of detection were 0.04 μg/g for DDT, 0.07 μg/g for DDE and 0.06 μg/g for DDD. In extractable fats from the above tissues and egg yolk, DDT and DDE were transferred/distributed throughout the tissues and egg yolk. DDD was detected only in the liver. The findings indicate that DDT is metabolized instantaneously to DDE/DDD in the hen's body and they are transferred rapidly into the egg‐forming tissues and egg yolk. Among the four tissues and yolk fats examined, the DDT levels were high in the ovary, oviduct and egg yolk; the DDE levels were high in the liver, ovary and oviduct and lowest in the yolk (P < 0.01).     

Cortical and Trabecular Bone Microstructure Did Not Recover at Weight‐Bearing Skeletal Sites and Progressively Deteriorated at Non‐Weight‐Bearing Sites During the Year Following International Space Station Missions

Risk for premature osteoporosis is a major health concern in astronauts and cosmonauts; the reversibility of the bone lost at the weight‐bearing bone sites is not established, although it is suspected to take longer than the mission length. The bone three‐dimensional structure and strength that could be uniquely affected by weightlessness is currently unknown. Our objective is to evaluate bone mass, microarchitecture, and strength of weight‐bearing and non‐weight‐bearing bone in 13 cosmonauts before and for 12 months after a 4‐month to 6‐month sojourn in the International Space Station (ISS). Standard and advanced evaluations of trabecular and cortical parameters were performed using high‐resolution peripheral quantitative computed tomography. In particular, cortical analyses involved determination of the largest common volume of each successive individual scan to improve the precision of cortical porosity and density measurements. Bone resorption and formation serum markers, and markers reflecting osteocyte activity or periosteal metabolism (sclerostin, periostin) were evaluated. At the tibia, in addition to decreased bone mineral densities at cortical and trabecular compartments, a 4% decrease in cortical thickness and a 15% increase in cortical porosity were observed at landing. Cortical size and density subsequently recovered and serum periostin changes were associated with cortical recovery during the year after landing. However, tibial cortical porosity or trabecular bone failed to recover, resulting in compromised strength. The radius, preserved at landing, unexpectedly developed postflight fragility, from 3 months post‐landing onward, particularly in its cortical structure. Remodeling markers, uncoupled in favor of bone resorption at landing, returned to preflight values within 6 months, then declined farther to lower than preflight values. Our findings highlight the need for specific protective measures not only during, but also after spaceflight, because of continuing uncertainties regarding skeletal recovery long after landing. © 2017 American Society for Bone and Mineral Research.     

Long‐term Effects of Dietary Anion–Cation Balance on Acid–Base Status and Bone Morphology in Reproducing Ewes

The beneficial effects of anionic salts on calcium metabolism have been shown by supplementing rations with such salts during the last 3 weeks of pre‐partum. However, there are few reports on the effects of anionic salts supplementation for periods of 4 weeks or longer on acid–base status, mineral metabolism and bone morphology. This study was conducted to evaluate the effects of the long‐term dietary supplementation of anionic salts on the acid–base status, plasma minerals concentrations and bone morphology in sheep. Twenty‐seven twin‐bearing sheep were assigned to two experimental groups and a control group, depending on dietary cation–anion difference (DCAD) (+272.6, −88.9 and +164.5 mEq/kg DM, respectively). Sheep assigned to each dietary treatment received their respective rations beginning 6 weeks pre‐partum and continuing until 12 days post‐partum. Diets containing anionic salts induced a mild metabolic hyperchloraemic acidosis from 1 week pre‐partum to 2 days post‐partum that was completely compensated by non‐respiratory mechanisms. These changes on acid–base status were accompanied by an increase of plasma ionized calcium levels. Plasma total calcium, phosphorus and magnesium concentrations were not affected by dietary treatment. Parathyroid hormone concentrations were related to the concentration of ionized calcium of plasma and were higher in sheep fed the cationic diet. Plasma osteocalcin levels were increased in sheep fed the anionic diet and cortical bone remodelling occurred in all the animals during late pregnancy in light and electron microscopy observation, but was particularly evident in the sheep fed the anionic diet. Bone turnover might be stimulated because of the role of the bone in buffering systemic acidosis. The data suggest that anionic salts ameliorated calcium metabolism around parturition by increasing bone resorption and the concentration of ionised calcium in plasma, possibly mediated by a mild hyperchloraemic metabolic acidosis induced by the salts.     

Changes in the Level of Endogenous Leptin,FSH, 17β‐Oestradiol and Metabolites during Lupin‐induced Increase in Ovulation Rate in Ewes

The aim of the study was to determine the changes in the plasma concentration of leptin during lupin feeding‐induced increase in the ovulation rate (OR) in ewes. Additionally, alterations in the plasma level of glycogenic amino acids and glucose (as the factors influencing leptin secretion) and the levels of follicle‐stimulating hormone (FSH) and 17β‐oestradiol (E‐2) (as the hormones regulated by leptin and engaged in recruitment, selection and development of ovulatory follicles) were analysed. Ninety‐six female Polish Lowland Sheep were used. All ewes were cyclic and synchronized with PGF_2α. The ewes were divided into two groups: control (n = 48), fed only with hay, and experimental (n = 48), received additionally lupin (Lupinus angustifolius) grain as a high‐protein and a high‐energy supplement. They were given lupin from the second to 13th day of the oestrous cycle at increasing doses (150–750 g/day per ewe). On the 11th day of cycle blood samples for analysis of hormones, amino acids and total glucose concentration, were collected from the jugular vein. OR was determined by laparoscopy of ovaries on the sixth day of the following oestrous cycle. Mean OR of ewes supplemented with lupin grain (1.687 ± 0.463) was 30.67% higher than that of control (1.291 ± 0.454). In spite of the unchanged body mass, a significant increase (P ≤ 0.05) in mean concentration of plasma leptin in the experimental ewes [2.17 ± 0.15 ng/ml human equivalent (HE)] was found in comparison with control (1.42 ± 0.12 ng/ml HE). A significantly (P ≤ 0.05) higher plasma FSH level in the ewes fed lupin (105.21 ± 5.87 ng/ml) compared with those fed hay (67.88 ± 6.03 ng/ml) was also found. However, plasma level of E‐2 decreased after lupin feeding. Moreover, in the ewes fed lupin the plasma concentrations of glucose and nine glycogenic amino acids (Gly, Ala, Val, Met, Leu, Ile, Tyr, Phe and Arg) were increased. It can be concluded that lupin feeding exerts the stimulatory effect on the OR in Polish Lowland Sheep. The increase in OR is connected with significantly higher plasma leptin level and coincident with rise in FSH, glycogenic amino acids and glucose concentration. In contrast, the level of plasma E‐2 was significantly decreased in lupin‐fed ewes.     

Galectins‐1 and ‐3 in Human Intervertebral Disc Degeneration: Non‐Uniform Distribution Profiles and Activation of Disease Markers Involving NF‐κB by Galectin‐1

Degeneration of the human intervertebral disc (IVD) is assumed to underlie severe clinical symptoms, in particular chronic back pain. Since adhesion/growth‐regulatory galectins are linked to arthritis/osteoarthritis pathogenesis by activating a pro‐degradative/‐inflammatory gene expression signature, we hypothesized a similar functional involvement of galectins in IVD degeneration. Immunohistochemical evidence for the presence of galectins‐1 and ‐3 in IVD is provided comparatively for specimens of spondylochondrosis, spondylolisthesis, and spinal deformity. Immunopositivity was detected in sections of fixed IVD specimens in each cellular compartment with age‐, disease‐, and galectin‐type‐related differences. Of note, presence of both galectins correlated with IVD degeneration, whereas correlation with age was seen only for galectin‐3. In addition, staining profiles for these two galectins showed different distribution patterns in serial sections, an indication for non‐redundant functionalities. In vitro, both galectins bound to IVD cells in a glycan‐dependent manner. However, exclusively galectin‐1 binding triggered a significant induction of functional disease markers (i.e., IL6, CXCL8, and MMP1/3/13) with involvement of the nuclear factor‐kB pathway. This study thus gives direction to further network analyses and functional studies on galectins in IVD degeneration. © 2019 The Authors. Journal of Orthopaedic Research® published by Wiley Periodicals, Inc. on behalf of Orthopaedic Research Society. J Orthop Res 37:2204–2216, 2019     

Plasma Concentrations of 15‐Ketodihydro‐PGF2α, Cortisol and Progesterone during Manual Twin Reduction in Thoroughbred Mares

The aim of the present study was to highlight the effect of two different techniques of one embryo crushing on some hormonal changes. Ten twinning mares were submitted to the mobile or fixed manual crushing of one blastocyst within day 19 after the last mating. Blood sample was collected from 20 min before to 90 min, 24 and 72 h after the procedure was performed to analyse 15‐ketodihydro‐PGF_2α, cortisol and progesterone plasma concentrations. Singleton pregnancy diagnosis was checked 72 h after crushing and at term of pregnancy. Because the unwanted crushing of both embryos occurred in one mare during the attempt of manual separation of the twins, that mare was not included in the evaluation of crushing‐induced hormonal changes. No significant differences in hormonal concentrations were observed after one embryo crushing and also when the effect of the mobile (n = 6) or fixed (n = 3) technique was specifically evaluated. When the effect of the two techniques on each post‐crushing sampling time hormonal levels was analysed, only a higher cortisol level 30 min after the fixed compared with the mobile technique was observed. The crushing performed within 19 days of gestation does not induce significant changes in 15‐ketodihydro‐PGF_2α, cortisol or progesterone plasma concentrations. When the fixed technique was performed, only a temporary higher cortisol concentration was seen 30 min after crushing, suggesting that the fixed technique might be responsible for a slight level of stress for the mare.     

Outbreaks of Pox Disease Due to Canarypox‐Like and Fowlpox‐Like Viruses in Large‐Scale Houbara Bustard Captive‐Breeding Programmes,in Morocco and the United Arab Emirates

Infectious diseases can be serious threats for the success of reinforcement programmes of endangered species. Houbara Bustard species (Chlamydotis undulata and Chlamydotis macqueenii), whose populations declined in the last decades, have been captive‐bred for conservation purposes for more than 15 years in North Africa and the Middle East. Field observations show that pox disease, caused by avipoxviruses (APV), regularly emerges in conservation projects of Houbara Bustard, despite a very strict implementation of both vaccination and biosecurity. Data collected from captive flocks of Houbara Bustard in Morocco from 2006 through 2013 and in the United Arab Emirates from 2011 through 2013 were analysed, and molecular investigations were carried out to define the virus strains involved. Pox cases (n = 2311) were observed during more than half of the year (88% of the months in Morocco, 54% in the United Arab Emirates). Monthly morbidity rates showed strong variations across the time periods considered, species and study sites: Four outbreaks were described during the study period on both sites. Molecular typing revealed that infections were mostly due to canarypox‐like viruses in Morocco while fowlpox‐like viruses were predominant in the United Arab Emirates. This study highlights that APV remain a major threat to consider in bird conservation initiatives.     

Metabolic Profiles and Bile Acid Extraction Rate in the Liver of Cows with Fasting‐induced Hepatic Lipidosis

This study was designed to monitor lipid profile in the portal and hepatic blood of cows with fasting‐induced hepatic lipidosis, and to compare the results with those in the jugular blood. The work was also carried out to investigate bile acid (BA) in these vessels, and further to investigate BA extraction rate in the liver. Five cows were equipped with catheters in the portal, hepatic and jugular veins (day 0), fasted for 4 days (day 1–day 4) and then refed (day 5–day 11). Before morning feeding, blood was sampled before, during and after fasting from the catheterized vessels. In the portal blood, the concentration of non‐esterified fatty acids (NEFA) showed a progressive increase and at day 5 there was an approximate twofold rise. Increased NEFA concentrations were also found similarly in the other two veins. At day 5, β‐hydroxybutyrate (BHBA) in the portal, hepatic and jugular blood rose to 197, 190 and 186% of the pre‐fasting value, respectively. However, the concentrations of NEFA and BHBA in the three veins gradually returned to pre‐fasting concentration during the refeeding period. Compared with the pre‐fasting value at day 0, the content of liver triglyceride (TG) increased significantly at day 5 (P < 0.01). In the liver, the hepatic extraction rate of BA dropped from 3.1 times pre‐fasting to 2.2 times during fasting. There were no significant differences in the concentrations of glucose, TG, total cholesterol, cholesterol esters, free cholesterol and phospholipids. The results of the current study show that metabolic alterations occur in the portal, hepatic and jugular veins during induction of hepatic lipidosis in cows, and mostly metabolites, with exception of BA concentration, run parallel. The decreased BA extraction rate in the liver of fasted cows was considered to reflect hepatic cell impairment caused by TG accumulation. Hopefully, the findings, at least in part, contribute to the explanation of the pathophysiology of hepatic lipidosis in dairy cows.     

Outcome of Patients with Preformed Donor‐Specific Antibodies Following Alemtuzumab Induction and Tacrolimus Monotherapy

It has been shown that low‐level preformed donor‐specific antibodies (DSAbs) detected by luminex beads in the setting of a negative CDC and flow cytometry crossmatch (CDC/FCXM) are associated with inferior allograft outcomes. The relevance of preformed DSAbs in patients receiving alemtuzumab induction and tacrolimus monotherapy has not been studied. Four hundred and eighty renal transplant recipients with a negative CDC/FCXM had their pretransplant sera retrospectively screened for DSAbs. 45/480 (9.4%) of patients were found to have preformed DSAbs. Females and patients receiving regrafts were more likely to have a DSAb (p = 0.008 and p < 0.0001, respectively). Patients with DSAbs had inferior allograft survival (p = 0.047), increased incidence of antibody‐mediated rejection (p < 0.0001) and inferior allograft function at 6 months posttransplant (p = 0.017). Patients with HLA class I DSAb (alone or in combination with a Class II DSAb) with high mean fluorescence intensities (MFIs) were at highest risk. We conclude that patients with preformed DSAb are at high risk of adverse outcomes when receiving a minimal immunosuppressive regime incorporating alemtuzumab induction. Patients found to have a preformed DSAb despite a negative crossmatch might benefit from augmented immunosuppression.     

Quantitation of cyclosporine‐sensitive and ‐resistant allospecific cytotoxic cells at birth

Abstract In the absence of clinically relevant models of acute rejection we have attempted to develop an assay to measure cyclosporine‐resistant allospecific cytotoxic cells in vitro, beginning at birth. The principle of limiting dilution analysis was applied to investigate umbilical cord bloods as responders. Responders were incubated for 1 h in different concentrations of cyclosporine and irradiated HLA mismatched stimulator cells from healthy adults added, followed by recombinant IL‐2. After 7 days, responders were tested against three europium‐labelled PHA blasts: stimulator, responder and third party. A significant number of cyclosporine‐resistant allospecific cytotoxic cell precursors were found in cord blood indicating prior activation. They may have been primed in utero against non‐inherited maternal HLA antigens. Cyclosporine‐resistant allospecific cytotoxic cell precursors were demonstrated in human umbilical cord blood using a quantitative assay. These cells may influence the reaction to subsequent transplants.     

Bile Acid Extraction Rate in the Liver of Cows Fed High‐Fat Diet and Lipid Profiles in the Portal and Hepatic Veins

The purposes of this study are to assess the responses of increased supplemental dietary fat in the cow, without upsetting rumen fermentation, on the bile acid (BA) extraction rate in the liver and to determine whether this diet would affect the postprandial lipid profiles in the portal and hepatic venous blood. Six Holstein cows were equipped with catheters fitted in the portal and hepatic veins. Two cows each were assigned randomly to a sequence of three dietary treatments of 21‐day period. The methodology of this study was based on the supplementation of the basal concentrate diet with 0 (control), 5, or 10% calcium salts of fatty acids (CSFA). The total bile acids were significantly increased in the portal and hepatic veins with the 5% CSFA diet, whereas no increase occurred with the 10% CSFA diet. Data obtained in this study showed that 10% CSFA diet failed to stimulate BA secretion to exceed the values obtained with 5% CSFA‐diet. Moreover, there was no change in the hepatic extraction rate of BA in animals fed either the 0 or 5% CSFA diets which ranged from 2.4 to 6.5‐fold and 3.1 to 7.3‐fold, respectively. However, the extraction rate increased sharply with the 10% CSFA diet (27‐fold). The median portal and hepatic concentrations of total lipids, triglycerides, total cholesterol, phospholipids and non‐esterified fatty acids did not show any significant increase during feeding of the control diet. Moreover, feeding either the 5 or 10% CSFA diet did not significantly increase these values in either vein.     

The sweets standing at the borderline between allo‐ and xenotransplantation

Animal cells are densely covered with glycoconjugates, such as N‐glycan, O‐glycan, and glycosphingolipids, which are important for various biological and immunological events at the cell surface and in the extracellular matrix. Endothelial α‐Gal carbohydrate epitopes (Galα3Gal‐R) expressed on porcine tissue or cell surfaces are such glycoconjugates and directly mediate hyperacute immunological rejection in pig‐to‐human xenotransplantation. Although researchers have been able to develop α1,3‐galactosyltransferase (GalT) gene knockout (KO) pigs, there remain unclarified non‐Gal antigens that prevent xenotransplantation. Based on our expertise in the structural analysis of xenoantigenic carbohydrates, we describe the immunologically significant non‐human carbohydrate antigens, including α‐Gal antigens, analyzed as part of efforts to assess the antigens responsible for hyperacute immunological rejection in pig‐to‐human xenotransplantation. The importance of studying human, pig, and GalT‐KO pig glycoprofiles, and of developing adequate pig‐to‐human glycan databases, is also discussed.