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a b s t r a c t The in vivo and post mortem performance and serum immunoglobulin G (IgG) concentration in kids born from goats fed conventional (group C) or genetically modified (group T) soy- bean meal were evaluated. The goat colostrum quality, in terms of chemical composition, as well as immunoglobulin concentration, and the presence of feed DNA fragments were also investigated. Kid birth weights were similar, while significantly (P < 0.05) higher in those born from goats in group C at day 30 and at slaughtering. In addition, kids from mothers fed conventional soybean recorded significant (P < 0.05) higher height at the withers and chest width. Concerning the post mortem measurements, only carcass weights were significantly affected by the treatment resulting in lighter T kids (P < 0.05). Colostrum from the treated groups recorded a significantly (P < 0.01) lower percentage of protein and fat. Similarly, both chemical parameters significantly differed in milk collected 15 days after kidding, although these differences disappeared in the successive samplings. Both colostrum and kids serum IgG concentration were significantly (P < 0.01) lower in the treated groups. Transgenic target DNA sequences (35S and CP4 EPSPS) were not detected in colostrum from goats that received a diet containing conventional soybean meal. By contrast, transgenic DNA fragments were amplified (P < 0.05) in samples from goats that received the transgenic soybean.

Aim of the present study was to evaluate the potential beneficial and/or adverse effects of Capsicum annuum L. (cv. Fiesta) extracts at two stage of ripening (immature and mature), and at two dosages (low and high) by evaluation of biochemical profile and oxidative status in CD-1 mice. The extracts were daily administered to mice by oral gavage for 20 days. At the end of the trial, the animals were euthanatized and blood was collected. Evidence of liver damage (increase of AST, ALT and bilirubin) in the group receiving the higher dosage of immature peppers extract were observed. Even if no adverse effects were seen at the lower doses, also no signs of beneficial effects in term of health status, biochemical profile and oxidative status were detected. These results are in contrast with in vitro findings and raise doubts about the possible use of Capsicum annuum L. (cv. Fiesta) as a nutritional supplement.

Simple Summary We evaluated the effects of a commercial nutraceutical supplementation in dogs naturally affected by Leishmania infantum. This nutraceutical supplement is proposed to be added to dog diet to foster physiological immune-response during chronic leishmaniasis. Parasite clearance is infrequently obtained by immune response in both the human and canine leishmaniasis. Pharmacological therapies frequently fail in the elimination of L. infantum that could have settled in deep organs, escaping immune responses and therapy. The conventional drug therapy improves clinical signs, reduces parasitemia as well as relapse of infection. This nutraceutical supplementation can improve the impaired immune response of the infected dogs to fight the disease. The use of nutraceuticals as immunomodulators in the treatment of visceral leishmaniasis has generated interest in the current approaches to treat the disease. In this clinical and immunological study, we investigated whether the administration of a nutritional supplement mediates the immune-modulatory response in canine leishmaniosis (CL) and improves the clinical outcome of the disease. With this purpose, we analysed T lymphocyte subsets in peripheral blood (PB) of 12 dogs naturally infected by Leishmania infantum, following treatment with a nutritional supplement. The regulatory T (Treg) cells and the T helper (Th) 1 population were specifically evaluated. The animals underwent complete clinical examination and blood sample collection for haematological, biochemical, serological and immunological analysis before treatment (T0), one month (T30) and 3 months (T90) after the onset of the nutraceutical supplementation. We observed that nutraceutical supplementation was associated with immunomodulation of Th1 response and significant clinical improvement of the animals. No side effects were observed. Therefore, a potential supportive role for the nutraceutical supplement during canine leishmaniasis is proposed.

Colon cancer is one of the major causes of cancer mortality worldwide. The analysed feeds, containing non-genetically modified (GM) soybean and Roundup Ready soybean, showed a different polyphenolic content and lipophilic composition. Non-GM soybean extract possessed twice the polyphenolic content of GM soybean and the highest number of sterols. Among them, γ-sitosterol was found to be the major constituent. Methanolic extract of non-GM soybean extract was more potent than GM soybean extract against colon carcinoma cell line LoVo using MTT assay, while the second one showed a slightly higher anti-inflammatory activity. The findings add to epidemiological evidence for the therapeutic effects of soy foods in colorectal carcinoma.

To investigate the effect of two insect meals (from Hermetia illucens, HI and Tenebrio molitor, TM larvae) on productive performance and blood profiles of Barbary partridge, ninety, seven days old partridges were divided into 5 groups (6 replicates, 3 partridges/replicate). Up to 64d, the groups fed 5 isoproteic and isoenergetic diets: the control fed a corn-soybean meal diet (SBM group); in TM25 and TM50 groups the 25 and 50% of SBM proteins were substituted by the protein from TM, respectively; in HI25 and HI50 groups the 25 and 50% of SBM were substituted by the protein from HI, respectively. The birds fed TM25 and both the HI levels reached a higher (P

The objective of the study was to evaluate the effect of passive transfer status, determined by measuring serum immunoglobulin (Ig) concentration 24 hours after parturition, on growth performance in buffalo calves allowed to nurse the dam during the first month of life. Serum Ig concentration 24 hours after birth ranged from 28.1 to 35.9 mg/ml, birth weight ranged from 29 to 41 kg, body weight 30 days after birth ranged from 48.5 to 62.9 kg. The Average Daily Gain (ADG) from birth to day 30 ranged from 448 to 1,089 g/d. Significant linear associations were detected between serum Ig concentration 24 hours after birth and day-30 weight (p< 0.05; R2 = 0.31) and between serum Ig concentration 24 hours after birth and ADG from birth to day 30 (p

A Cre/loxP-based fate mapping approach was used to follow the regions of the mouse thyroid labeled by the serotonin transporter SERT. Reporter gene expression (lacZ) is activated by Cre expression from the SERT locus in SERT(Cre/+) ;ROSA26R compound mouse embryos. Cell labeling, first detected in the thyroid primordium at the E10.5 prenatal stage, was followed until the postnatal day P30. The co-localization of lacZ staining in the same cells that express the transcription factors Nkx2.1 and Pax8 at the E12.5 stage confirms their identity as thyroid cell precursors. SERT immunohistochemistry on thyroid sections of E18.5 embryos showed SERT expression in thyroid follicular cells. Western blotting analysis confirmed the expression of the protein in adult thyroid tissue and cultured FRTL-5 cells. These results describe the fate of SERT-expressing cells during thyroid development, suggesting an active role of SERT in the development and functions of mammalian thyroid. They also highlight the possibility to use the SERT-Cre mouse line as a good Cre driver in early thyroid development.

This study aimed to investigate the antidiabetic, antilipidaemic and antioxidant activities of Citrus medica cv Diamante (Rutaceae) hydroalcoholic (CD) peel extract in Zucker diabetic fatty (ZDF) rats. The ability of CD to protect against oxidative stress was investigated by using different in vitro assays and in vivo by using the reactive oxygen metabolites-derived compounds (d-ROMs) test and the biological antioxidant potential test (BAP). Two different doses of CD extract (300 and 600 mg/kg/die) were administered at ZDF rats for 4 weeks. CD reduced cholesterol and triglycerides levels. A dose-dependent effect on body weight and serum glucose levels was observed. A decrease of d-ROMs and an increase of BAP were recorded by using the dose of 600 mg/kg. The extract inhibited lipid peroxidation (IC50 value of 0.23 mg/ml). These findings suggest as an efficient phytotherapeutic approach in combating hyperlipidaemic and hyperglycaemic disorders.

The presence of DNA fragments in tissues from rabbits given genetically modified (GM) soya-bean meal (solvent extracted) was investigated by using the polymerase chain reaction (PCR) approach. Moreover, the possible effects on cell metabolism were evaluated by determination of several specific enzymes in serum, heart, skeletal muscle, liver and kidney. The chloroplast sequence for tRNA Leu by using the Clor1/Clor2 primers designed on chloroplast trnL sequence was clearly detected. On the contrary, two couples of species specific primers for conventional (Le1-5/Le 1-3 which amplifies the soya bean lectin gene) and genetically modified (35S1/35S2 which amplifies the 35S CMV promoter that is present in the genomic structure of GM soya bean) soya bean were not found in all samples. No differences in enzyme levels were detected in serum, but a significant increase of lactic dehydrogenase, mainly concerning the LDH1 isoenzyme was found in particular in kidney and heart but not in the muscle, thus suggesting a potential alteration in the local production of the enzyme. Finally, no significant differences were detected concerning body weight, fresh organ weights and no sexual differences were detected.

Aim of this study was to investigate the effects of supplementing dairy goat diets with hydrogenated palm oil (HPO; 50 g/day/head) on milk yield and composition, and on milk fatty acids profile and Stearoyl-CoA desaturase gene expression. Compared with the control diet (C), supplementation with HPO increased milk yield and fat percentage. By contrast, milk protein percentage was not different between groups. Supplementation with HPO increased (P < 0.01) short and medium chain fatty acids, as well as C14:0 and C16:1 cis-9 acids (P < 0.05). Milk from control group showed significantly (P < 0.01) higher concentration of C18:0 and lower (P < 0.05) of cis-9 trans-11CLA. Concerning the desaturase index, the HPO addition sig- nificantly affected C16:1/C16:0 and cis-9 trans-11CLA/C18:1 trans-11 (P < 0.01) and C18:1/C18:0 (P < 0.05) ratios. The SCD gene expression (arbitrary units) was higher, even if not significantly, in the control com- pared to the treated group and in both groups decreased along the trial (May: 0.55 vs 0.41; June: 0.51 vs 0.38; July: 0.50 vs 0.37 for C and T group, respectively). Concerning health properties of milk, showing beneficial (CLAs and palmitoleic acid increase) and adverse (myristic acid increase) effect, our results suggest that further studies are needed to address a risk/benefit assessment of HPO supplementation in goats.

The presence of DNA fragments in blood and milk from goats fed conventional (control) or Roundup Ready® soybean meal solvent extracted (s.e.; treated) was investigated by using a polymerase chain reaction approach. The same investigation was carried out on blood, skeletal muscle and organs from kids of both groups fed only dams' milk until weaning. Moreover, the possible effects on cell metabolism were evaluated by determination of several specific enzymes in serum, heart, skeletal muscle, liver and kidney. Fragments of the multicopy chloroplast (trnL) gene were found in blood and milk samples from goats of both groups. In kids, the chloroplast fragments were found in samples of both groups. In samples, which proved positive for the presence of chloroplast DNA, fragments of the specific soybean single copy gene (lectin) were detected in several blood and milk samples. The same fragment was also found in control and treated groups of kids. Transgenic fragments were not found in those samples, which were found positive for chloroplast fragments of control groups of either goats or kids. On the contrary, in blood and milk of treated goats, fragments both of the 35S promoter and the CP4 epsps gene were detected. These fragments were also found in treated kids with a significant detection of the 35S promoter in liver, kidney and blood, and of the CP4 epsps gene fragment in liver, kidney, heart and muscle. A significant increase in lactic dehydrogenase, mainly concerning the lactic dehydrogenase-1 isoenzyme was found in heart, skeletal muscle and kidney of treated kids, thus suggesting a change in the local production of the enzyme. Finally, no significant differences were detected concerning kid body and organ weight.

Two days before kindling, 228 New Zealand White rabbit does were homogeneously divided into two groups (114 does per group) and fed the same diet. After delivery, the litters were equalized to 8 pups. From 1 to 35 days of age (weaning), the control group (CONT) did not receive any treatment while in the experimental group (LAC) the nests were sprayed with a commercial product containing lyophilized Lactobacillus plantarum dissolved in water (12 g/L). L. plantarum was sprayed on the litters (5 mL per rabbit) once a day during seven consecutive days after delivery. After one week of rest, the treatment was repeated for another week according to the same experimental protocol. Mortality rate, recorded on all the litters (912 rabbits per group) was significantly lower in the LAC group (9.9 vs 17.2%; Pin vivo performance of the 24 litters per group, and rabbits of both groups reached a similar weight at weaning (938 vs 932 g for LAC and CONT groups, respectively). Rabbits from the LAC group showed fermentative activity of caecal microflora (total volatile fatty acids 24.8 vs 14.5 mmol/L; Pvs 63.9% of total white blood cells; PL. plantarum (1.25x106 CFU/g). It might be supposed that the changes in caecal microflora can affect our results and improve the sanitary status of Lactobacillus-sprayed rabbits in the period 1-35 days of age.

This study aimed to evaluate the total phenolic and flavonoid content, radical scavenging activity (by DPPH and ABTS tests) and antioxidant capacity (by β-carotene bleaching test) of Annona cherimola (cherimoya) fruits cultivated in Italy for human consumption. The metal chelating activity and ferric reducing power were also determined. A. cherimola peel and pulp were characterized by a total phenolic content of 14.6 and 12.6 mg chlorogenic acid equivalents/100 g fresh weight, respectively. A similar trend was observed with flavonoid content. Both extracts exhibited high antioxidant activity through different mechanisms of action. In particular, peel extract demonstrated the strongest free radical scavenging activity with an IC 50 value of 57.7 μg/mL. The same extract was more effective in preventing β-carotene oxidation (IC 50 value of 63.5 μg/mL after 60 min) and showed higher chelating ability (IC 50 value of 79.6 μg/mL) than pulp extract. This work demonstrated the high quality of cherimoya fruits cultivated in Italy, and recommends the peel of this fruit product that may be of interest from a functional point of view as a major source of natural antioxidants.

Hepatocyte Growth Factor (HGF)/c-MET signaling has an emerging role in promoting cell proliferation, survival, migration, wound repair and branching in a variety of cell types. HGF plays a crucial role as a mediator of stromal–epithelial interactions in the normal prostate but the precise biological function of HGF/c-Met interaction in the normal prostate and in prostate cancer is not clear. HGF has two naturally occurring splice variants and NK1, the smallest of these HGF variants, consists of the HGF amino terminus through the first kringle domain. We evaluated the intracellular signaling cascades and the morphological changes triggered by NK1 in human prostate epithelial cell line PNT1A which shows molecular and biochemical properties close to the normal prostate epithelium. We demonstrated that these cells express a functional c-MET, and cell exposure to NK1 induces the phosphorylation of tyrosines 1313/1349/1356 residues of c-MET which provide docking sites for signaling molecules. We observed an increased phosphorylation of ERK1/2, Akt, c-Src, p125FAK, SMAD2/3, and STAT3, down-regulation of the expression of epithelial cell–cell adhesion marker E-cadherin, and enhanced expression levels of mesenchymal markers vimentin, fibronectin, vinculin, α-actinin, and α-smooth muscle actin. This results in cell proliferation, in the appearance of a mesenchymal phenotype, in morphological changes resembling cell scattering and in wound healing. Our findings highlight the function of NK1 in non-tumorigenic human prostatic epithelial cells and provide a picture of the signaling pathways triggered by NK1 in a unique cell line.

The serotonin transporter (SERT) gene has been proposed as a candidate gene responsible for the sudden infant death syndrome (SIDS). In this study, for the first time we obtained a SERT-knockout (KO) mouse model which reproduces SIDS phenotype. SERT-KO mice were generated by mating SERT(Cre/+) heterozygous mice. The SERT-KO mouse embryos at the pre-natal stage E18.5 were lacking of SERT mRNA and protein expression in the heart. A premature death of 75% of SERT-KO mice occurred in the first week after birth. LacZ staining of whole mounts and tissue sections of the heart from SERT(Cre/+);ROSA26R adult mice and E18.5 embryos demonstrated a marked localized expression of SERT in the right ventricle, the conal region, the vasculature, the atrial septum, the ventricular valves, and the sinoatrial node of the conduction system. These data suggest a cardiac phenotype for the sudden death of SERT-KO mice. Histological analysis of heart sections showed that SERT-KO mice develop cardiac fibrosis. Increased collagen accumulation in the myocardium and the valvular and perivascular regions, and enhanced expression of alpha-smooth muscle actin were detected in the heart of SERT-KO mice versus wild-type (WT) mice. Interestingly, higher expression levels of the 5-HT2A receptor and increased levels of phospho-SMAD2/3 and phospho-ERK1/2 were detected in SERT-KO mouse heart versus WT mice. Overall, our findings provide i) new insights into the role of SERT gene in SIDS, and ii) the first in vivo validation of the molecular mechanism involving the activation of TGF-beta1 signalling in the cardiac fibrosis.

Multiple evidence demonstrate a role for serotonin and its transporter SERT in heart valve development and disease. By utilizing a Cre/loxP system driven by SERT gene expression, we recently demonstrated a regionally restricted distribution of SERT-expressing cells in developing mouse heart. In order to characterize the cell types exhibiting SERT expression within the mouse heart valves at early developmental stages, in this study we performed immunohistochemistry for Islet1 (Isl1) and connexin-43 (Cx-43) on heart sections from SERT Cre/+;ROSA26R embryos previously stained with X-gal. We observed the co-localization of LacZ staining with Isl1 labelling in the outflow tract, the right ventricle and the conal region of E11.5 mouse heart. Cx-43 labelled cells co-localized with LacZ stained cells in the forming atrioventricular valves. These results demonstrate the cardiomyocyte phenotype of SERT-expressing cells in heart valves of the developing mouse heart, thus suggesting an active role of SERT in early heart valve development. © 2008 Elsevier Inc. All rights reserved.

Expression of the serotonin transporter (SERT) gene during mouse development L. M. Pavone & P. Mithbaokar & V. Mastellone & R. Lo Muto & A. Spina & V. Maharajan & G. Paino & L. Avallone Published online: 7 August 2008 # Springer Science + Business Media B.V. 2008

Choroid plexuses (CPs) play pivotal roles in a wide range of processes that establish, survey, and maintain the biochemical and cellular status of the central nervous system. Mammalian CPs contain a very high density of serotonin receptors, and serotonin has been shown to affect CP functions. The serotonin transporter (SERT) regulates the entire serotonergic system, including serotonin receptors by means of modulation of serotonin concentration in the extracellular fluid. In this study, the expression of SERT in the CPs from the brain of a mammalian species, Bubalis bubalis, was established. By immunogold labeling in scanning electron microscopy, SERT immunoreactivity was found to be localized on the apical surface of the choroid epithelium. In particular, SERT positivity was detected on the apical portion of villi, and both on the membrane and in the cytoplasm of grouped cells on the surface of the choroid epithelium. Significantly, no SERT was detected in blood vessels irrigating the CPs. The expression of SERT mRNA transcripts of 440 bp in the CPs was detected by reverse-transcription polymerase chain reaction, and Western blotting analysis revealed the presence of three isoforms of the protein with molecular masses of approximately 70, 80, and 140 kDa, respectively, probably corresponding to differently glycosylated SERT. Our findings provide the first report of SERT detection in the CPs of buffalo brain and indicate that this protein is locally synthesized from the choroid epithelial cells. We suggest that SERT might have an important role in mammalian CPs, possibly regulating the serotonin flow between brain and rest of the body.