Short communication: Associations between blood fatty acids, ß-hydroxybutyrate, and α-tocopherol in the periparturient period in dairy cows: An observational study.

The objective of the present study was to examine the relationships between blood concentrations of fatty acids, ß-hydroxybutyrate (BHB), and α-tocopherol during the periparturient period in dairy cows. Blood samples were collected from 131 cows belonging to 4 different commercial dairy farms in southeastern Europe (Greece and Italy). We determined blood concentrations of fatty acids, BHB, and α-tocopherol at dry-off, at calving, and 30d postpartum. Results indicated that fatty acid concentrations were low at dry-off, reached maximum value at calving, and then declined at 30d postpartum. In fact, fatty acid concentrations at 30d postpartum were 50% lower than at calving. In contrast, BHB concentrations were low at dry-off, increased by 27% at calving, and continued to increase by another 20% at 30d postpartum. Overall, we found a weak correlation between fatty acids and BHB throughout the periparturient period. Concentrations of α-tocopherol were lowest at calving, and we detected no differences in α-tocopherol concentrations at dry-off or 30d postpartum. Negative correlations between fatty acids and α-tocopherol were highly significant at 30d postpartum and approached the level of significance at dry-off. However, both correlations became nonsignificant following the adjustment of α-tocopherol with cholesterol, indicating that the correlations were a reflection of changes in lipid transport. We found significant negative correlations (strong at dry-off and weak at 30d postpartum) between BHB and α-tocopherol after adjustment with cholesterol. The physiological basis for the negative correlations between BHB and α-tocopherol, especially that at dry-off, is not known and should not be taken to imply a cause-effect relationship. However, it opens the door to investigating the effects of vitamin E on liver function in dairy cows.

The effects of superoxide dismutase-rich melon pulp concentrate on inflammation, antioxidant status and growth performance of challenged post-weaning piglets.

Piglets can often suffer impaired antioxidant status and poor immune response during post-weaning, especially when chronic inflammation takes place, leading to lower growth rates than expected. Oral administration of dietary antioxidant compounds during this period could be a feasible way to balance oxidation processes and increase health and growth performance. The aim of the trial was to study the effects of an antioxidant feed supplement (melon pulp concentrate) that contains high concentration of the antioxidant superoxide dismutase (SOD) on inflammation, antioxidant status and growth performance of lipopolysaccharide (LPS) challenged weaned piglets. In total, 48 weaned piglets were individually allocated to four experimental groups in a 2×2 factorial design for 29 days. Two different dietary treatments were adopted: (a) control (CTR), fed a basal diet, (b) treatment (MPC), fed the basal diet plus 30 g/ton of melon pulp concentrate. On days 19, 21, 23 and 25 half of the animals within CTR and MPC groups were subjected to a challenge with intramuscular injections of an increasing dosage of LPS from Escherichia coli (serotype 0.55:B5) (+) or were injected with an equal amount of PBS solution (-). Blood samples were collected at the beginning of the trial and under the challenge period for interleukin 1ß, interleukin 6, tumour necrosis factor α, haptoglobin, plasma SOD activity, total antioxidant capacity, reactive oxygen species, red blood cells and plasma resistance to haemolysis, and 8-oxo-7, 8-dihydro-2'-deoxyguanosine. Growth performance was evaluated weekly. A positive effect of melon pulp concentrate was evidenced on total antioxidant capacity, half-haemolysis time of red blood cells, average daily gain (ADG) and feed intake, while LPS challenge increased pro-inflammatory cytokines and haptoglobin serum concentrations, with a reduced feed intake and gain : feed (G : F). The obtained results show that oral SOD supplementation with melon pulp concentrate ameliorates the total antioxidant capacity and the half-haemolysis time in red blood cell of post-weaning piglets, with positive results on growing performance.

Influence of estrus of dairy goats on somatic cell count, milk traits, and sex steroid receptors in the mammary gland.

Two experiments were conducted to study the effect of the stage of a spontaneous estrus cycle on milk yield and constituents [somatic cell count (SCC), fat, protein, caseins, lactose, and urea content] and on estrogen receptor-alpha (ERalpha ) and progesterone receptor (PR) immunostaining in the mammary gland. In experiment I, the major components of milk and SCC were monitored weekly in 80 lactating Saanen goats for 6 wk, whereas detection of estrus was daily. In experiment II, milk samples were collected daily for SCC determination during 1 spontaneous estrus (d 0) until the second spontaneous estrus in 14 Saanen goats. The day of the estrous cycle was confirmed by plasma progesterone and 17beta-estradiol levels. Immunoreactivity of ERalpha and PR was analyzed in mammary gland samples of 8 Saanen goats (d 0, n = 4; d 10, n = 4) and the number of positive nuclei and intensity of the staining were evaluated in 1,000 cells. In experiment I, milk casein and protein percentages were significantly affected by the stage of estrous cycle; during proestrus and estrus, these variables were higher (3.32 +/- 0.06 and 4.44 +/- 0.08) than during metestrus (3.03 +/- 0.07 and 4.07 +/- 0.10), but not higher than during diestrus (3.23 +/- 0.06 and 4.35 +/- 0.09, respectively). In experiment II, daily measurement of SCC revealed higher levels at estrus (7,195 +/- 672 x 10(3) cells/mL) and a decline toward the luteal phase (1,694 +/- 672 +/- 10(3) cells/mL). Estrogen receptor-alpha and PR immunostaining were exclusively detected on epithelial cells. The percentage of positive nuclei to ERalpha was higher on d 0 than on d 10 (75.4 +/- 8.8 vs. 68.3 +/- 8.8%), but no change was observed for PR (4.0 +/- 0.3 vs. 3.5 +/- 0.4%). The average immunostaining intensity for both receptors was greater on d 0 than on d 10 (ERalpha : 1.44 +/- 0.02 vs. 1.35 +/- 0.02; PR: 0.079 +/- 0.008 vs. 0.057 +/- 0.008). The high SCC at estrus in experiment II was associated with high plasma estradiol and low progesterone, suggesting that the increased SCC could be brought about by the estrogen-induced proliferation and exfoliation of epithelial cells. In addition, this action may be supported by the higher sensitivity to estrogens (ERalpha content) found at d 0.

UCP1 and UCP2 expression in different subcutaneous and visceral adipose tissue deposits in 30 days old goat kids and effect of fatty acid enriched diets.

Beside its importance in the first hours of life, brown adipose tissue has also significant roles in the following stages of growth and in adults by regulating energy metabolism, but its identification in adult ruminants is still controversial. Quantitative PCR, followed by histological confirmation, was used to investigate UCP expression and brown and white adipocytes' distribution in 30-day-old goat kids. The influence of maternal diet enriched with either fish oil or stearic acid was investigated as well. Results showed the differential expression of both UCP1 and UCP2 genes between subcutaneous and visceral adipose tissues, suggesting a different thermogenic activity between the two macro areas. The maternal diet influenced neither UCP1 nor UCP2 gene expression. The presence of multilocular adipocytes in 1-month goat kids is remarkable, as suggests thermogenic activity in non-newborn animals. Further insights into characteristics and functions of adipose tissue in young and adult goats are worth exploring.

Functional interaction between benzodiazepine and GABA recognition sites in aged rats.

The present study was undertaken to explore whether there may be age-related changes in benzodiazepine binding and in the functional interaction between GABA and benzodiazepine recognition sites. Data indicate an increase in benzodiazepine binding sites with age. Moreover the functional interactions between GABA and benzodiazepine receptor sites are differentially affected by aging. GABA is less active in enhancing benzodiazepine binding in the older animals because of the loss of GABA receptors, and Diazepam may be more active in enhancing GABA receptor binding in the aged animals because there are more benzodiazepine receptors in this group. An understanding of the relevance of the apparent alteration in coupling between GABA and benzodiazepine receptors must permit a better definition of the behavioral manifestation of their biochemical phenomenon.

Comparative efficacy of cyclandelate versus flunarizine in the prophylactic treatment of migraine.

In a double-blind, parallel-group randomised trial of 3 months' duration, the efficacy of cyclandelate 800 mg twice daily in migraine prophylaxis was compared with that of flunarizine 5mg daily in 40 patients. In comparison with placebo and baseline values, both drugs significantly relieved symptoms of migraine as assessed by indices of pain total index, headache index, analgesic consumption and number of migraine days. Patients taking flunarizine experienced side effects such as drowsiness, weight gain and asthenia, while the most common complaint reported with cyclandelate was gastric upset. These results suggest that cyclandelate may be a useful alternative in migraine prophylaxis.

The influence of different fibrous supplements in the diet on ruminal histology and histometry in veal calves.

The aim of this study was to determine whether the administration of four different solid feeds would influence selected morphological and morpho-functional aspects of the rumen mucosa in veal calves. The fibrous supplementation of the liquid diet of veal calves has been provided by recent EU formulation (EC Council Directive 91/629/1991; EC Council Directive 97/2/1997). Twenty-five Holstein calves were assigned to either exclusively liquid diet (milk replacer, control), or pelleted feed, corn silage, extruded feed, dried corn silage. The morpho-functional effects of the fibre-containing diets were examined evaluating histological and histometrical characteristics of ruminal mucosa after the slaughter of calves. There were slight to severe histological abnormalities in the rumens of all animals examined. The severe histological abnormalities were present in calves given pelleted feed, corn silage, and extruded feed. Dried corn silage caused less ruminal damage. We found that the length and epithelial thickness of ruminal papillae were higher in control veal calves than in dietary fibre-supplemented animals. The results of the present study, even if partially, support the EU prescription in the use of fibre diets in veal calves as integration of the traditional milk replacer diet.

Structural patterns of swine ileal mucosa following L-glutamine and nucleotide administration during the weaning period. An histochemical and histometrical study.

Dietary supplementations with L-glutamine and/or nucleotides were screened for their effects on intestinal mucosa in 16 female weaning piglets. The animals were transported to the university's facilities 24 hours after weaning. They were grouped four to a pen in controlled environmental conditions and fed one of the following four diets for 28 days: control diet (C); C+0.5% L-glutamine (G); C+0.05% "nucleotides" (N); and C+0.5 % L-glutamine+0.05% "nucleotides" (GN). Individual body weights and feed intake per group were recorded at the beginning and the end of the study as well as weekly during it. There were no significant performance differences among the groups. After 28 days the animals were slaughtered and the distal ileum and liver were examined histologically. Anti-proliferating cell nuclear antigen (PCNA) as well as anti-human macrophage immunostaining, and a modified TdT-mediated dUTP nick-end labeling technique (TUNEL) were performed, and intraepithelial lymphocyte percentage was evaluated to assess morpho-functional aspects of the ileum. Histometry was performed by assessing cell indices and counts of immuno-reactive structures. Feeding G and/or N resulted in an increase in villi (V) height, crypt (C) depth, and a decrease in V:C ratio (P<0.01). In addition, feeding G and/or N resulted in an increase in mitotic mucosal cells (M), and a decrease in apoptotic mucosal cells (A), thus decreasing the A:M index (P<0.01). The percentages of mucosal macrophages were greater in G and/or N groups (P<0.001) than in control piglets, and similarly among the groups the percentages of intraepithelial lymphocytes varied (P<0.01). Our data showed that the diet supplementation with G and/or N had positive effects on some morpho-functional characteristics of piglet ileal mucosa. These ameliorative effects may potentially be linked to a good responsiveness of piglets to a stressful period, like a precocious weaning is in this species.

Regulation of GABA receptor binding to synaptic plasma membrane of rat cerebral cortex: the role of endogenous phospholipids.

Triton X-100 treatments produced an extensive depletion of proteins and phospholipids and a marked increase of [3H] GABA binding on synaptic plasma membranes (SPM). Maximal [3H]GABA binding was obtained with three Triton X-100 treatments (+ 174% with respect to control). Phospholipase C, which removes only the phospholipid polar head, induces a 40% increase of [3H]GABA binding only after treatments resulting in extensive protein depletion. In reconstitution experiments phosphatidylethanolamine, the largest phospholipid removed, induced a 30-35% inhibition of [3H]GABA binding in Triton X-100 treated membranes; in contrast phosphatidylserine and phosphatidylcholine did not produce significant changes. The reconstitution of phospholipase C-treated SPM preparations with exogenous phosphatidylethanolamine, phosphatidylserine, phosphatidylcholine or phosphoethanolamine and 1,2-dipalmitoylglycerol, products of phospholipase C activity, did not yield significant changes. This evidence, which argues against a direct role of phospholipids on the regulation of GABA binding, should, however, suggest that the GABA binding component of the receptor site is a lipoprotein or a lipid-depending protein.

Chronic GM1 ganglioside treatment reduces dopamine cell body degeneration in the substantia nigra after unilateral hemitransection in rat.

The effect of GM1 ganglioside on the recovery of dopaminergic nigro-striatal neurons was studied in rats after unilateral hemitransection. GM1 treatment partially prevented the decrease of tyrosine hydroxylase (TH) activity caused by hemitransection in the substantia nigra ipsilateral to the lesion. Concomitantly a significant increase of TH-immunoreactivity in the substantia nigra was also detected. In particular, chronic treatment with GM1 prevented the disappearance of TH-positive cell bodies in the substantia nigra and induced the appearance of longer TH-positive dendrites with respect to the saline treatment. These data indicate that GM1 treatment maintains the number of dopaminergic cell bodies in the substantia nigra after hemitransection by protecting against retrograde neuronal degeneration.

Monosialoganglioside internal ester stimulates the dopaminergic reinnervation of the striatum after unilateral hemitransection in rat.

The effects of the administration of GM1 monosialoganglioside internal ester, AGF2, on the dopaminergic reinnervation of the striatum in rats with unilateral hemitransection has been studied. AGF2 increases the apparent Vmax and the density of tyrosine-hydroxylase-positive nerve terminals in the striatum of the lesioned side, without modification of the tyrosine-hydroxylase activity in the unlesioned side. AGF2, at lower doses, is more active than its parent natural molecule GM1. AGF2 has a larger half-life and a higher distribution volume than GM1, and undergoes a slow hydrolysis in the serum releasing the original natural compound GM1. Mannitol and dexamethazone, often used to prevent swelling of the brain after injury, or isoaxonine, proposed to stimulate neurite growth are unable to reproduce the effects of AGF2 on the recovery of striatal tyrosine-hydroxylase activity after hemitransection. The data are compatible with the view that AGF2, through its conversion into GM1, facilitates the collateral sprouting of the nigro-striatal dopaminergic neurons.

In rats, the metabotropic glutamate receptor-triggered hippocampal neuronal damage is strain-dependent.

The effect of intrahippocampal (i.h.) and intraocular (i.o.) administration of the selective metabotropic glutamate receptor (mGluR) agonist (1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid (1S,3R-ACPD) was studied in different rat strains. A massive hippocampal damage was observed in CD/SD and Fischer 344 but not in SD/Rij and Brown Norway rats 7 days following the i.h. injection of 1S,3R-ACPD, while no retinal damage was observed following its i.o. administration. Moreover, 1S,3R-ACPD reduced the N-methyl-D-aspartate (NMDA) toxicity in the retina of both CD/SD and SD/Rij rats. Regardless of its toxic action on hippocampal neurons the i.h. injection of 1S,3R-ACPD caused an acute stimulation of motor activity in both CD/SD and SD/Rij rats. This effect was blocked by the intracerebroventricular (i.c.v.) administration of the putative mGluR antagonist L-2-amino-3-phosphono-propionic acid (L-AP3). It is suggested that the differential expression of mGluR subtypes might determine their role in brain pathology.

Effects of gangliosides on the functional recovery of damaged brain.

The effect of GM1 ganglioside on the recovery of dopaminergic nigro-striatal neurons was studied in rats after unilateral hemitransection. GM1 treatment favoured the collateral sprouting of dopaminergic axons in the striatum as indicated by the induced increase of tyrosine hydroxylase (TH) activity and immunofluorescence. Concomitantly GM1 partially prevented the decrease of TH activity caused by the hemitransection in the substantia nigra ipsilateral to the lesion. A significant increase of TH immunoreactivity was also detected in the substantia nigra: GM1 prevented the disappearance of TH-positive cell bodies and increased the formation of TH-positive collaterals and dendrites with respect to the saline treatment. The addition of GM1 to embryonic dissociated mesencephalic cell cultures stimulates the expression of dopaminergic characteristics as suggested by the increase of 3H-DA uptake.

[Cost methodologies in dialysis in relation to the Italian National Health Service]. / Metodologie dei costi in dialisi in rapporto all'azienda salute italiana.

This work analyses the economic aspects of dialysis in Italy in relation to government resources allocated to the health service in general. The authors illustrate the procedures used to estimate the resources required by the dialytic programme. The costs of dialytic programmes in different cities and at different periods in the history of the Italian health service are compared. A concrete example is outlined of the economic management of dialysis and the authors demonstrate how the results were obtained using cost analysis.

Computer image analysis: an additional tool for the identification of processed poultry and mammal protein containing bones.

The aims of this study were (1) to evaluate the potential of image analysis measurements, in combination with the official analytical methods for the detection of constituents of animal origin in feedstuffs, to distinguish between poultry versus mammals; and (2) to identify possible markers that can be used in routine analysis. For this purpose, 14 mammal and seven poultry samples and a total of 1081 bone fragment lacunae were analysed by combining the microscopic methods with computer image analysis. The distribution of 30 different measured size and shape bone lacunae variables were studied both within and between the two zoological classes. In all cases a considerable overlap between classes meant that classification of individual lacunae was problematic, though a clear separation in the means did allow successful classification of samples on the basis of averages. The variables most useful for classification were those related to size, lacuna area for example. The approach shows considerable promise but will need further study using a larger number of samples with a wider range.

The chemical code of porcine enteric neurons and the number of enteric glial cells are altered by dietary probiotics.

BACKGROUND: The enteric nervous system (ENS) contains chemically coded populations of neurons that serve specific functions for the control of the gastrointestinal tract. The ability of neurons to modify their chemical code in response to luminal changes has recently been discovered. It is possible that enteric neuronal plasticity may sustain the adaptability of the gut to changes in intestinal activity or injury, and that gut neurons may respond to an altered intestinal environment by changing their neuropeptide expression. METHODS: We used immunohistochemical methods to investigate the presence and localization of several neuronal populations and enteric glia in both the small (ileum) and large (cecum) intestine of piglets. We assessed their abundance in submucosal and myenteric plexus from animals treated with the probiotic Pediococcus acidilactici compared with untreated controls. KEY RESULTS: The treated piglets had a larger number of galanin- and calcitonin gene-related peptide (CGRP)-immunoreactive neurons than controls, but this was limited to the submucosal plexus ganglia of the ileum. Moreover, immunohistochemistry revealed that glial fibrillary acidic protein-positive enteric glial cells were significantly higher in the inner and outer submucosal plexuses of treated animals. CONCLUSIONS & INFERENCES: The neuronal and glial changes described here illustrate plasticity of the ENS in response to an altered luminal environment in the gastrointestinal tract.

Effects of protected fish oil in the diet of periparturient dairy goats on phenotypic variation in blood and milk leukocytes.

The goal of this study was to evaluate the effects of dietary protected fish oil (FO) on phenotypic variation in blood, milk leukocytes, and some productive and metabolic parameters in periparturient dairy goats. About 12 Alpine goats, selected from a larger group of second-parity animals, were fed from 15 days before kidding until the 15th day of lactation with the same basal diet that had been supplemented with either 47 g/head per day of FO or 47 g/head per day hydrogenated palm oil (PO). Dry matter intake, live body weight (LBW), body condition score (BCS), and productive performance were evaluated in 2 weeks after kidding. On days 15, 7, and 2 before kidding and days 2, 7, and 15 after kidding, plasma samples were collected for evaluation of alanine aminotransferase, aspartate aminotransferase, non-esterified fatty acids, glucose, beta-hydroxybutyrate, cholesterol, and urea levels. White blood cell and blood leukocyte subsets were counted in whole blood samples on the kidding day, as well as at 1, 4, and 15 days after kidding. In addition, milk somatic cell count, intramammary infection (IMI), and milk leukocyte subsets were evaluated on days 4 and 15 after kidding. No differences were observed in dry matter intake and BCS, while LBW was higher in FO-fed animals. Milk production and composition, plasma metabolites, and liver enzymes were similar in both experimental groups. Blood CD4 positive cells increased constantly (P = 0.05) in FO-fed group, while CD8 and CD14 cell counts significantly increased 4 days after kidding (P < 0.01). Milk leukocyte subsets showed a significant (P < 0.01) decrease in PO-fed group and a non-significant increase (P = 0.34) in FO-fed group, despite the presence of coagulase negative staphylococci IMI. The results of the productive performance evaluation agreed with those of many other studies, which did not find any significant differences between dairy goats fed diets enriched with FO or PO supplements. The administration of FO to dairy goats in transition appeared to affect the variation in blood leukocytes with a constant increase in CD4- and CD8-positive cells in comparison with a PO fat-supplemented diet.