Transcriptomic landscape of TIMP3 oncosuppressor activity in thyroid carcinoma.

BACKGROUND: Papillary thyroid cancer (PTC) is the most frequent thyroid tumor. The tissue inhibitor of metalloproteinase-3 (TIMP3) gene encodes a matrix metalloproteinases inhibitor that exerts a tumor suppressor role in several tumor types. TIMP3 is frequently downregulated in PTC by promoter methylation. We have previously functionally demonstrated that TIMP3 exerts an oncosuppressor role in PTC: TIMP3 restoration in the PTC-derived NIM1 cell line affects in vitro migration, invasion and adhesive capability, while reduces tumor growth, angiogenesis and macrophage recruitment in vivo. To get a deeper insight on the mediators of TIMP3 oncosuppressor activity in thyroid tumors, here we focused on the TIMP3 related transcriptome. METHODS: TCGA database was used for investigating the genes differentially expressed in PTC samples with low and high TIMP3 expression. Genome wide expression analysis of clones NIM1-T23 (expressing a high level of TIMP3 protein) and NIM1-EV (control empty vector) was performed. Gene sets and functional enrichment analysis with clusterProfiler were applied to identify the modulated biological processes and pathways. CIBERSORT was used to evaluate the distribution of different immunological cell types in TCGA-PTC tumor samples with different TIMP3 expression levels. Real time PCR was performed for the validation of selected genes. RESULTS: Thyroid tumors with TIMP3-high expression showed a down-modulation of inflammation-related gene sets, along with a reduced protumoral hematopoietic cells fraction; an enrichment of cell adhesion functions was also identified. Similar results were obtained in the TIMP3-overexpessing NIM1 cells in vitro model, where a down-regulation of immune-related function gene sets, some of which also identified in tumor samples, was observed. Interestingly, through enrichment analysis, were also recognized terms related to cell adhesion, extracellular matrix organization, blood vessel maintenance and vascular process functions that have been found modulated in our previous in vitro and in vivo functional studies. CONCLUSIONS: Our results highlight the correlation of TIMP3 expression levels with the regulation of inflammatory functions and the immune infiltration composition associated with different PTC prognosis, thus providing a broader view on the oncosuppressor role of TIMP3 in PTC.

The Special Case of the Spectral Emission of a Tb3+ Mono Metal Complex.

The fine structure in the spectral lines of the visible fluorescence of Tb3+ complexes are replaced by a single peak in the case of a singular molecular complex Tb(H3 PTC)3 , where H4 PTC represents perylene-3,4,9,10-tetracarboxylic acid, and its emission wavelength depends on the film thickness. This single peak challenges the old creed that the f-orbital electrons of Tb3+ are always protected from the influence of the surrounding atoms. We perform density functional theory calculations to show that the wavefunction of the ground state is localized and in addition, spin-polarized, and this facilitates fluorescent transitions under UV to the first excited state instead of the fundamental state. We discuss the possibility of making a spintronic device with the molecule, Tb(H3 PTC)3 .

All-perylene-derivative for white light emitting diodes.

An organic-based bright white light emitting compound, namely Tb(H3PTC)3 [H4PTC = perylene-3,4,9,10-tetracarboxylic acid], able to be used as part of a white diode and as a part of a RGB system that can withstand high temperatures (∼700 K), is developed using perylene-3,4,9,10-tetracarboxylic dianhydride (PTCDA) and terbium(iii) nitrate pentahydrate as precursors by hydrothermal synthesis. Using PTCDA as the red emitter and the new derivative of it, Tb(H3PTC)3, as the blue-green emitter, along with a common deep blue LED can form a RGB system for display technologies, around room temperature. Temperature-dependent photoluminescence properties of the Tb(H3PTC)3 compound are also investigated for the involved excitonic-emission processes and the respective recombination lifetimes. The terbium(iii) complex was prepared using a procedure that is reproducible, easily modifiable, inexpensive, and environmentally friendly, opening new pathways for its large-scale applications. Unlike PTCDA, Tb(H3PTC)3 has been shown to be soluble in N-methyl-2-pyrrolidone (NMP) as well as in dilute aqueous solutions of this organic solvent in a straightforward procedure. The light emission properties are intimately correlated with the molecular structure and electronic properties of Tb(H3PTC)3 elucidated by experimental results of X-ray Absorption Near Edge Spectroscopy (XANES), Extended X-ray Absorption Fine Structure (EXAFS) and Density Functional Theory (DFT) calculations. A bright fluorescence yield is attained with a small amount of material either in solution or in solid form showing its potential to be used in state-of-the-art organic optoelectronic devices.

Histology, composition, and quality traits of chicken Pectoralis major muscle affected by wooden breast abnormality.

Only a few years ago, the poultry industry began to face a recent abnormality in breast meat, known as wooden breast, which frequently overlaps with white striping. This study aimed to assess the impact of wooden breast abnormality on quality traits of meat. For this purpose, 32 normal (NRM), 32 wooden (WB), and 32 wooden and white-striped (WB/WS) Pectoralis major muscles were selected from the same flock of heavy broilers (males, Ross 708, weighing around 3.7 kg) in the deboning area of a commercial processing plant at 3 h postmortem and used to assess histology, proximate (moisture, protein, fat, ash, and collagen) and mineral composition (Mg, K, P, Na and Ca), sarcoplasmic and myofibrillar protein patterns, and technological traits of breast meat. Compared to the normal group, WB/WS fillets showed more severe histological lesions characterized by fiber degeneration, fibrosis, and lipidosis, coupled with a significantly harder texture. With regard to proximate and mineral composition, abnormal samples exhibited significantly (P < 0.001) higher moisture, fat, and collagen contents coupled with lower (P < 0.001) amounts of protein and ash. Furthermore, increased calcium (131 vs. 84 mg kg(-1); P < 0.05) and sodium (741 vs. 393 mg kg(-1); P < 0.001) levels were found in WB/WS meat samples. The SDS-PAGE analysis revealed a significantly lower amount of calcium-ATPase (SERCA, 114 kDa), responsible for the translocation of Ca ions across the membrane, in normal breasts compared to abnormal ones. As for meat quality traits, fillets affected by wooden abnormality exhibited significantly (P < 0.001) higher ultimate pH and lower water-holding/water-binding capacity. In particular, compared to normal, abnormal samples showed reduced marinade uptake coupled with increased drip loss and cooking losses as well. In conclusion, this study revealed that meat affected by wooden breast or both wooden breast and white striping abnormalities exhibit poorer nutritional value, harder texture, and impaired water-holding capacity.

Excitatory and inhibitory enteric innervation of horse lower esophageal sphincter.

The lower esophageal sphincter (LES) is a specialized, thickened muscle region with a high resting tone mediated by myogenic and neurogenic mechanisms. During swallowing or belching, the LES undergoes strong inhibitory innervation. In the horse, the LES seems to be organized as a "one-way" structure, enabling only the oral-anal progression of food. We characterized the esophageal and gastric pericardial inhibitory and excitatory intramural neurons immunoreactive (IR) for the enzymes neuronal nitric oxide synthase (nNOS) and choline acetyltransferase. Large percentages of myenteric plexus (MP) and submucosal (SMP) plexus nNOS-IR neurons were observed in the esophagus (72 ± 9 and 69 ± 8 %, respectively) and stomach (57 ± 17 and 45 ± 3 %, respectively). In the esophagus, cholinergic MP and SMP neurons were 29 ± 14 and 65 ± 24 vs. 36 ± 8 and 38 ± 20 % in the stomach, respectively. The high percentage of nitrergic inhibitory motor neurons observed in the caudal esophagus reinforces the role of the enteric nervous system in the horse LES relaxation. These findings might allow an evaluation of whether selective groups of enteric neurons are involved in horse neurological disorders such as megaesophagus, equine dysautonomia, and white lethal foal syndrome.

Relationship between pectoralis major muscle histology and quality traits of chicken meat.

A trial was conducted to evaluate the influence of myodegeneration of pectoralis major muscle on quality traits and chemical composition of breast meat of heavy-size male broilers. For this purpose, a total of 72 pectoralis major muscles were randomly collected from broilers farmed under homogeneous conditions and graded into three categories (mild, n=22; moderate, n=33; and severe, n=17) based on the presence of abnormal fibers (giant fibers, fibers with hyaline degeneration, and damaged and/or necrotic fibers) evaluated by histological and immunohistochemical analysis. Color, pH, drip loss, Allo-Kramer shear values, and chemical composition (moisture, proteins, total lipids, ashes, and collagen) were determined on nonmarinated breast meat. Purge loss and cook loss, total yield, and Allo-Kramer shear values were measured on vacuum-tumbled samples. Samples showing moderate myodegeneration had the highest mean cross-sectional area of the fibers, while samples with severe myodegeneration had myofibers of different diameter and without the characteristic polygonal shape, multifocal degeneration and necrosis, as well as infiltration of CD3-immunoreactive cells. Cooking losses of nonmarinated meat were lower in the mild group with respect to moderate and severe groups (21.4 vs. 24.7 and 24.7%; P<0.001). Breast muscles with severe damage, in comparison with mild degenerated samples, showed higher moisture (75.4 vs. 74.4%; P<0.05) and lower protein percentages (21.1 vs. 22.6%; P<0.001). The lipid percentage of severely degenerated samples was higher than that from moderate group (2.94 vs. 2.36; P<0.05), while collagen content was not modified by histological lesion levels. Marinated meat from the mild group had higher uptake and total marinade yield after cooking. In conclusion, almost all breast fillets of heavy broiler chickens produced under intensive farming systems had histological lesions, which reflected on the chemical composition of the meat and the impaired water holding/binding capacities of the meat.

Effect of fasting and refeeding on expression of genes involved in the gastric nutrient sensing and orexigenic control of pigs.

Knowledge on orexigenic signals in the pig stomach is poor. Gastric amino acid sensing by taste receptor type 1 member 3 (T1R3) and calcium-sensing receptor (CASR), and active ghrelin release, controlled by preproghrelin, proprotein convertase (PC1/3) and ghrelin O-acyltransferase (GOAT) genes, may be affected by fasting or refeeding. Twelve pigs (12.0 kg LW) were adapted to a base diet and assigned to three individual feeding schedules: Control (C), fed twice a day; Fasting (F), fasted for 24 h; Refeeding (R), fasted for 24 h and refed before slaughtering. Gastrointestinal segments were collected for histology and molecular biology analyses. Total RNA isolated from oxyntic and pyloric mucosae was reverse transcribed, specific porcine primers were designed and transcript quantification was performed by real-time RT-PCR. F decreased villus height in duodenum (p < 0.01) and ileum (p < 0.05) vs. C and R. R increased oxyntic PC1/3 (p < 0.05) and tended to increase oxyntic preproghrelin (p = 0.06), and pyloric GOAT (p = 0.07) gene expression vs. C. PC1/3 gene expression was higher in pyloric mucosa. Ghrelin-positive cells numbers were not different between the two gastric mucosae. Gastrin expression tended to be higher in R than in C and F (p = 0.068 and p = 0.055). CASR was higher in pyloric than in oxyntic mucosa, and pyloric CASR expression tended to be higher in R than in C (p = 0.072). T1R1 was not affected by treatment. Our results indicate that the pool of genes involved in the secretion of active ghrelin is active both in oxyntic and pyloric mucosa of pig. Refeeding can significantly affect the expression of genes that control octanoyl-ghrelin production and partially the amino acid sensing by CASR gene, while the absence of effect of fasting on the expression of ghrelin-related genes needs further confirmations.

Effect of added dietary threonine on growth performance, health, immunity and gastrointestinal function of weaning pigs with differing genetic susceptibility to Escherichia coli infection and challenged with E. coli K88ac.

Threonine (Thr) is important for mucin and immunoglobulin production. We studied the effect of added dietary Thr on growth performance, health, immunity and gastrointestinal function of weaning pigs with differing genetic susceptibility to E. coli K88ac (ETEC) infection and challenged with ETEC. Forty-eight 24-day-old weaned pigs were divided into two groups by their ETEC susceptibility using mucin 4 (MUC4) gene as a marker (2 MUC4(-/-) , not-susceptible, and 2 MUC4(+/+) , susceptible, pigs per litter). Within genotype, pigs were fed two different diets: 8.5 (LThr) or 9.0 (HThr) g Thr/kg. Pigs were orally challenged on day 7 after weaning and slaughtered on day 12 or 13 after weaning. Before ETEC challenge, HThr pigs ate more (p < 0.05). The diet did not affect post-challenge growth, but HThr tended to increase post-challenge feed efficiency (p = 0.087) and overall growth (p = 0.087) and feed efficiency (p = 0.055). Before challenge, HThr pigs excreted less E. coli (p < 0.05), while after challenge, diet did not affect the number of days with diarrhoea and ETEC excretion. MUC4(+/+) pigs responded to the challenge with more diarrhoea, ETEC excretion and anti-K88 IgA in blood and jejunal secretion (p < 0.001). HThr pigs had a higher increase of anti-K88 IgA values in jejunal secretion (p = 0.089) and in blood (p = 0.089, in MUC4(+/+) pigs only). Thr did not affect total IgA and IgM values, morphometry of jejunum, goblet cells count in colon, total mucin from jejunum and colon, but varied jejunal goblet cells counts (p < 0.05). In the first two post-weaning weeks, 8.5 g Thr/kg diet may be not sufficient to optimize initial feed intake, overall feed efficiency and intestinal IgA secretion and to control the gut microbiota in the first post-weaning week, irrespective of the pig genetic susceptibility to ETEC infection.

Graphene­boron nitride superlattices: the role of point defects at the BN layer.

We investigate, by means of first-principles calculations, the role of hBN point defects on the energetical stability and electronic structure of heterostructures composed of graphene atop hBN, rotated at angles of 13.17°, 9.43° and 7.34°. We consider, as possible point defects, boron and nitrogen vacancies and antisites, substitutional oxygen at the nitrogen site ON, substitutional carbon dimers, and nitrogen interstitials. The electronic and structural properties of all defects were analyzed. Among these, the most stable is ON, with negative formation energies at several possible rotation angles and chemical environments. Under such conditions, ON doping can raise the Fermi level of the neutral system by as much as 1 eV relative to graphene's Dirac point, reaching the band crossing between adjacent Dirac cones at the M point of the heterostructure Brillouin zone. This could lead to interesting electronic transport properties without the need for electrostatic doping.

Generation of induced pluripotent stem cells (CSSi017-A)(12862) from an ALS patient carrying a repeat expansion in the C9orf72 gene.

Genetic expansions of the hexanucleotide repeats (GGGGCC) in the C9orf72 gene appear in approximately 40% of patients with familial ALS and 7% of patients with sporadic ALS in the European population, making this mutation one of the most prevalent genetic mutations in ALS. Here, we generated a human induced pluripotent stem cell (hiPSC) line from the dermal fibroblasts of a patient carrying a 56-repeat expansion in an ALS disease-causing allele of C9orf72. These iPSCs showed stable amplification in vitro with normal karyotype and high expression of pluripotent markers and differentiated spontaneously in vivo into three germ layers.

MRI versus conventional measures of disease activity and structural damage in evaluating treatment efficacy in juvenile idiopathic arthritis.

OBJECTIVE: To compare the American College of Rheumatology paediatric (ACRp) response criteria and conventional radiography with MRI findings in a cohort of patients with juvenile idiopathic arthritis. METHODS: Forty consecutive patients (30 girls, 10 boys; median age 10.8 years) with arthritis of the wrist starting treatment with disease-modifying antirheumatic drugs or biological agents were recruited. At 1-year follow-up the treatment response was assessed by ACRp criteria and radiographic progression using the adapted Sharp/van der Heijde method. Wrist MRIs were evaluated using both the paediatric-MRI and the OMERACT rheumatoid arthritis MRI scores. Sensitivity to change of clinical and imaging variables was assessed by standardised response mean (SRM) and relative efficiency (RE) was used to compare SRMs. RESULTS: ACRp90 responders showed a significantly higher decrease in MRI synovitis score (median change -4) than non-responders (median change 0), ACRp30-50 responders (median change 0) and ACRp70 responders (median change -1) (p=0.0006, Kruskal-Wallis test). Non-responders showed significantly higher radiographic progression than ACRp90 responders (pB=0.016). The MRI synovitis score showed a greater responsiveness to change (SRM 1.69) compared with the majority of ACR core set of variables. MRI erosion scores were less responsive than conventional radiography in detecting destructive changes (RE <1). MRI follow-up revealed no signs of inflammation in four out of 24 wrists with clinically inactive disease. CONCLUSION: Only ACRp90 responders showed a significant decrease in synovitis and the halting of structural damage, suggesting that levels of response higher than ACRp30 are more appropriate for assessing drug efficacy. The excellent responsiveness of MRI and its ability to detect subclinical synovitis make it a promising outcome measure.

Comparison of breast muscle traits and meat quality characteristics in 2 commercial chicken hybrids.

A trial was conducted to compare muscle traits and meat quality characteristics of the pectoralis muscle in 2 chicken commercial hybrids having standard (SBY) and high breast yield (HBY), respectively. A total of 2,124 one-day-old male chicks, equally divided into 2 experimental groups represented by strains (SBY and HBY), were grown using homogenous conditions and fed the same standard diets until reaching live weight of 4.2 kg at 53 and 55 d for the SBY and HBY groups, respectively. Thirty-six birds per each genotype were randomly selected, and their pectoralis major muscles were used to assess meat quality properties (color attributes, pH, drip loss, cook loss, Allo-Kramer shear values after cooking, moisture, proteins, total lipids, and ashes) as well as histological traits (cross-sectional area, frequency of abnormal fibers, and intramuscular fat infiltration). As expected, HBY genotype had higher breast yield (31.0 vs. 30.0%; P ≤ 0.05). Histological evaluations showed that HBY pectoralis muscles had higher cross-sectional fiber area coupled with a dramatically higher (P ≤ 0.001) incidence of abnormal fibers and more abundant infiltration of intramuscular fat. Moreover, histopathological anomalous features such as central nuclei, proliferation of endomysial and perimysial collagen, inflammatory infiltrate, and necrosis of the fibers were also observed. As for meat quality, SBY hybrid showed lower ultimate pH values (5.97 vs. 6.07; P ≤ 0.01), whereas overall color parameters were not affected by genotype. Breast meat from the HBY genotype also exhibited significantly lower ability to retain liquid during refrigerated storage (drip loss, 2.46 vs. 2.06%; P ≤ 0.05) and cooking (26.2 vs. 21.1%; P ≤ 0.05) as well as higher shear-force values (2.59 vs. 2.11 kg/g; P ≤ 0.001). Finally, with regard to chemical composition, significant differences (P ≤ 0.05) were detected in protein (22.8 vs. 23.5%) and lipid (1.65 vs. 1.82%) contents, which were significantly lower in the HBY hybrid, whereas moisture content tended (P = 0.07) to be inferior in the SBY hybrid.

Blend of natural and natural identical essential oil compounds as a strategy to improve the gut health of weaning pigs.

Weaning is one of the most critical phases in pig's life, often leading to postweaning diarrhoea (PWD). Zinc oxide (ZnO), at pharmacological doses, has been largely used to prevent PWD; however, due to antimicrobial co-resistant and environmental pollution issues, the EU banned its use in June 2022. Natural or natural identical components of essential oils and their mixture with organic acids are possible alternatives studied for their antimicrobial, anti-inflammatory and antioxidant abilities. This study aimed to evaluate the effect of two blends of natural or natural identical components of essential oils and organic acids compared to ZnO on health, performance, and gut health of weaned pigs. At weaning (d0), 96 piglets (7 058 ± 895 g) were assigned to one of four treatments balanced for BW and litter: CO (control treatment), ZnO (2 400 mg/kg ZnO from d0 to d14); Blend1 (cinnamaldehyde, ajowan and clove essential oils, 1 500 mg/kg feed); Blend2 (cinnamaldehyde, eugenol and short- and medium-chain fatty acids, 2 000 mg/kg feed). Pigs were weighed weekly until d35. Faeces were collected at d13 and d35 for microbiota (v3-v4 regions of the 16 s rRNA gene) and Escherichia coli (E. coli) count analysis. At d14 and d35, eight pigs/treatment were slaughtered; pH was recorded on intestinal contents and jejunal samples were collected for morphological and gene expression analysis. From d7-d14, the Blend2 had a lower average daily gain (ADG) than CO and ZnO (P < 0.05). ZnO and Blend1 never differed in ADG and feed intake. At d14, ZnO had a lower caecum pH than all other treatments. The CO treatment had a higher abundance of haemolytic E. coli than Blend1 (P = 0.01). At d13, the ZnO treatment had a lower alpha diversity (P < 0.01) and a different microbial beta diversity (P < 0.001) compared to the other treatments. At d13, the ZnO treatment was characterised by a higher abundance of Prevotellaceae_NK3B31_group (Linear Discriminant Analysis (LDA) score = 4.5, P = 0.011), Parabacteroides (LDA score = 4.5, P adj. = 0.005), the CO was characterised by Oscillospiraceae UCG-005 (LDA score = 4.3, P adj. = 0.005), Oscillospiraceae NK4A214_group (LDA score = 4.2, P adj. = 0.02), the Blend2 was characterised by Megasphaera (LDA score = 4.1, P adj. = 0.045), and Ruminococcus (LDA score = 3.9, P adj. = 0.015) and the Blend1 was characterised by Christensenellaceae_R-7_group (LDA score = 4.6, P adj. < 0.001) and Treponema (LDA score = 4.5, P adj. < 0.001). In conclusion, Blend1 allowed to maintain the gut health of postweaning piglets through modulation of the gut microbiome, the reduction of haemolytic E. coli while Blend2 did not help piglets.

Metastable phase formation and structural evolution of epitaxial graphene grown on SiC(100) under a temperature gradient.

Multilayer epitaxial graphene was obtained from a 6H-SiC(001) substrate subjected to a temperature gradient from 1250 to 1450 °C. Scanning tunneling microscopy and x-ray diffraction were used to identify the structure and morphology of the surface, from which the formation of a metastable phase was inferred. By a comparison between microscopy and diffraction data, we report the appearance of misoriented Si-doped graphene in cold regions (1250 °C) of the substrate. This metastable phase occurs in domains where silicon sublimation is incomplete and it coexists with small domains of epitaxial graphene. At 1350 °C this phase disappears and one observes complete graphene-like layers (although misoriented), where rotational registry between the underlying epitaxial graphene and additional layers is absent. At 1450 °C the stacking among layers is established and the formation of highly oriented single crystalline graphite is complete. The stability of this Si-rich metastable phase at 1250 °C was confirmed by first-principles calculations based on the density functional theory.

Nanometrological porphyrins.

A new cationic silver N-alkylpyridylporphyrin complex is able to 'sense' nanometric conductive particles with a diameter below 10 nm. The luminescence of the molecule changes its maximum from red to blue when it embraces a conductive (metallic or semiconducting) nanoparticle. The change is explained on the basis of a charge transfer between the molecule and the conductive nanoparticle along with a geometrical distortion of the porphyric ring and pyridinium substituents. This new molecule could be used to sense nanoparticle contamination in the environment, in the industry of heterogeneous catalysis and many other branches of nanometrological applications.

Anomalous response of supported few-layer hexagonal boron nitride to DC electric fields: a confined water effect?

We use electric force microscopy (EFM) to study the response of supported few-layer hexagonal boron nitride (h-BN) to an electric field applied by the EFM tip. Our results show an anomalous behavior in the dielectric response of h-BN atop Si oxide for different bias polarities: for a positive bias applied to the tip, h-BN layers respond with a larger dielectric constant than the dielectric constant of the substrate, while for a negative bias, the h-BN dielectric constant appears to be smaller. Based on ab initio calculations, we propose that this behavior is due to a water layer confined between the Si oxide substrate and h-BN layers. This hypothesis was experimentally confirmed by sample annealing and also by a comparative analysis with h-BN on a non-polar substrate.

Effect of chronic heat stress on gastrointestinal histology and expression of feed intake-regulatory hormones in broiler chickens.

Heat stress (HS) dramatically impairs the growth performance of broiler chickens, mainly as a consequence of reduced feed intake due to the loss of appetite. This study was aimed at evaluating the alterations induced by chronic HS conditions on the morphological and morphometric features of the gastrointestinal (GI) tract and on the expression of some enteroendocrine cells (EECs) involved in the regulation of feed intake in chickens. Three hundred male chickens (Ross 308) were divided into two experimental groups and raised either in thermoneutral environment for the whole fattening period (0-41 days) (TNT group) or subjected to chronic HS conditions (30 °C for 24 h/day) from 35 to 41 days (HS group). Samples of proventriculus, duodenum, jejunum and cecum were collected from 24 broilers (12/group). Haematoxylin-eosin was used for the morphometric evaluations, while immunohistochemistry was applied for the evaluation of EECs expressing ghrelin (GHR), cholecystokinin (CCK), neuropeptide Y (NPY), glucagon-like peptide-1 (GLP-1), and serotonin (5-HT). In the proventriculus, HS reduced total wall thickness and mucous layer height (P ≤ 0.01) as well as mean diameter, circumference, and area of the compound tubular glands (P ≤ 0.001) with respect to TNT. The small intestine of HS birds was characterised by decreased villous height and total thickness (duodenum, P ≤ 0.01; jejunum, P ≤ 0.001), whereas crypt depth and width were reduced only in the jejunum (P ≤ 0.01). HS had negligible effects on the morphological aspects of the cecum. In the proventriculus, an increase in GHR and NPY EECs was observed in response to HS (P ≤ 0.001). Similarly, the small intestine villi of the HS group showed greater GLP-1 (P ≤ 0.05), 5-HT (P ≤ 0.001) and CCK (P ≤ 0.01) EECs. Moreover, the expression of 5-HT EECs was higher in the duodenal (P ≤ 0.01) and jejunal (P ≤ 0.01) crypts of HS birds, whereas GLP-1 and CCK EECs increased only in jejunal crypts (P ≤ 0.05). Finally, 5-HT EEC expression was increased in the cecum of HS group (P ≤ 0.01). In conclusion, these outcomes demonstrate that chronic HS induces morphometric alterations not only in the small intestine but also in a key organ such as the proventriculus. Furthermore, HS conditions affect the presence and distribution of EECs, suggesting that some GI peptides and biogenic amine may be implicated in the regulation of appetite and voluntary feed intake in heat-stressed broiler chickens.

Effect of an Escherichia coli F4/F18 bivalent oral live vaccine on gut health and performance of healthy weaned pigs.

Oral live vaccines stimulate host immunity, but they could also affect intestinal mucosa development and gut microbiota of piglets during the postweaning. The aim of this study was to determine the effect of an oral vaccine against Escherichia coli F4 and F18 (Coliprotec F4/F18®), on gut functionality and integrity, growth performance and health status of postweaning piglets. A total of 96 weaned piglets (23.30 ±â€¯1.85 days of age; 7334 ±â€¯1039 g BW) were divided into two groups (16 replicates/group; three piglets/replicate) as follows: (1) Control (CO), fed a standard diet (prestarter up to 14 days, then starter feed); (2) Treated (TRT): as CO but vaccinated with Coliprotec F4/F18® at weaning (day 0). Piglets were weighed at day 0 and weekly until day 35. Individual faecal score was recorded daily. Piglets were sacrificed at days 10 (1/3 of total) and 35 (2/3). Samples of jejunum mucosa and of cecum content were collected for morphometric, immunohistochemistry analysis and for microbiota profile analysis, respectively. Data were fitted using a linear model including treatment, class of starting BW as fixed factors and litter as random factor. From days 0 to 7, piglets from the TRT group tended to have a higher average daily gain (+22.6%, P = 0.08) and average daily feed intake compared to the CO group (+13.2%, P = 0.022). Gain to feed ratio was lower in the TRT group from days 14 to 35 (-6.6%, P = 0.011). From days 7 to 14, the TRT group had a higher diarrhoea index (-199%, P < 0.001). Crypt depth was higher in the CO group (+10.9%, P = 0.04) at day 10, but not at day 35. Jejunal expression of Claudin-4 (probability of having a score = 3) was higher in the TRT group at day 10 (CO = 1.50% vs TRT = 2.69%, P < 0.0001) and day 35 (CO = 1.29% vs TRT = 1.92%, P = 0.012). Oral vaccine affected beta diversity at day 10 (P = 0.040; R2 = 0.05) and increased the abundance of specific taxa and genera in the cecum at day 10, including Prevotella (lg2FC = 23.2, FDR < 0.001). The results showed how an Escherichia coli-based vaccine supplied to weaned pigs can promote gut health by controlling symptoms of the postweaning perturbation in the first 2 weeks postweaning. In addition, the vaccine strains showed a probiotic-like effect by modulating gut microbiota favouring the establishment of beneficial bacteria, and by promoting gut barrier integrity.

Accumulation of Selected Trace Elements in Shads from Three Lakes: First Insights from Italian Pre-Alpine Area.

The investigation of trace element pollution is important for the environmental assessment and management of lacustrine ecosystems, especially when these represent critical freshwater resources in densely populated areas. In this context, this study determined the levels of 15 trace elements in muscles of shad, Alosa agone (Scopoli 1786), a commercialized zooplanktivourous fish, from three primary, but currently still poorly studied, Italian lakes, namely, Lake Como, Iseo, and Garda. Research findings show that shads present similar trace element accumulation patterns among lakes, except for arsenic, which occurs at lower levels in Lake Como. Results provide evidence also for mercury biomagnification in fish, whereas all the other selected trace elements undergo bio-dilution through the same trophic chain. Maximum allowable limits for foodstuff were exceeded for chromium and selenium in shads, whereas mercury levels exceeded the European Environmental Quality Standard biota. These results highlight the need for regular monitoring activities of trace elements in the biota of these lakes.

Maternal antibiotic treatment affects offspring gastric sensing for umami taste and ghrelin regulation in the pig.

BACKGROUND: Scarce is knowledge on the process regulating the development of acid secretion, orexigenic signaling, and chemosensing in the stomach of young pigs. Changes of early microbial encounters by suckling pigs can interact with the gut maturation, by the induction of different molecular signaling. Our goal was to assess if the age of offspring and the maternal environment, influenced by sow antibiotic treatment peripartum, could affect gastric morphology and the expression of genes involved in the control of hydrochloric secretion, feed intake, taste, and inflammation in offspring stomach. METHODS: 84 pigs from sows fed a diet with amoxicillin (on -d10 to +d21 from farrowing, ANT) or without (CON) were sacrificed at d14, d21, d28 (weaning) or d42. Samples of oxyntic (OXY), pyloric (PY) and cardiac mucosae close to OXY were collected and parietal and enteroendocrine cells (EECs) were counted. Relative gene expression of a set of 11 key genes (ATP4A, SSTR2, GAST, GHRL, MBOAT4, PCSK1, GNAT1, TAS1R1, TAS1R3, IL8 and TNF) was assessed by qRT-PCR. In addition, 40 offspring obtained from the same ANT and CON sows were offered a normal or a fat-enriched diet for 4 weeks between 140 and 169 d of age, and then OXY and PY were sampled. RESULTS: The number of parietal and EECs increased with age (P < 0.001). ATP4A increased with age (within suckling, P = 0.043, post-weaning vs. suckling, P < 0.001), SSTR2 increased only after weaning (P < 0.001). In OXY, GHRL increased during suckling (P = 0.012), and post-weaning as a trend (P = 0.088). MBOAT4 tended to increase during suckling (P = 0.062). TAS1R1 increased from suckling to post-weaning period (P =0.001) and was lower in ANT offspring (P = 0.013). GNAT1 in PY was higher in ANT offspring (P = 0.041). Antibiotic treatment of sows peripartum increased expression of GHRL and MBOAT4 in OXY of growing-finishing offspring aged 5 months. CONCLUSIONS: Data show that sensing for umami taste and ghrelin regulation can be affected by maternal environment, but the development of acid secretion, orexigenic signaling and taste perception in the stomach are mostly developmentally controlled.