Cholecystokinin like immunoreactivity in the brains of young Meishan and Duroc pigs(4)

SUMMARY: Cholecystokinin (CCK), a peptide found in both the gastrointestinal tract and brain, has been shown to be involved in the control of feed intake in a variety of animals including the pig. Chinese breeds of pigs such as the Meishan are noted for slow growth and heavy adipose deposition. In this study we have described the regional cholecystokinin-like immunoreactivity (CCK-IR) concentrations in the brain of young Duroc and Meishan pigs utilizing radioimmunoassay. Brains of days 1, 10, and 20 postnatal pigs from each breed were examined. The CCK-IR increased with age in all three areas examined (cortex, medulla, and hypothalamus). The cortical concentrations rose significantly from days 1 to 10 and from days 10 to 20. The levels in the hypothalamus and medulla increased significantly between days 1 and 20. There were no statistically significant differences in CCK-IR between the breeds at any of the three ages examined. Our results indicate that a rise in CCK-IR in the regions of the brain involved in the control of feed intake may parallel the ability of the young pigs to assimilate nutrients from a solid diet. ZUSAMMENFASSUNG: Cholecystokinin-?hnliche Immunreaktivit?t in den Gehirnen junger Meishan- und Durocschweine Das Peptid Cholecystokinin (CCK) wird im Gastrointestinaltrakt und im Gehirn gefunden und beeinflu?t Futteraufnahme in einer Reihe von Tieren einschlie?lich Schwein. Chinesische Rassen wie Meishan sind wegen ihres langsamen Wachstums und der starken Fettablagerung bekannt. In dieser Studie beschreiben wir regionale Cholecystokinin-?hnliche Immunreaktivit?ts-(CCK-IR)Konzentrationen im Gehirn junger Duroc- und Meishantiere, mittels Radioimmunassay bestimmt. Gehirne von 1, 10 und 20 Tage alten Ferkeln jeder Rasse wurden untersucht. CCK-IR nahm mit dem Alter in allen drei untersuchten Organen zu (Kortex, Medulla und Hypothalamus). Die kortikalen Spiegel stiegen vom Tag 1 bis 10 und vom Tag 10 bis 20 signifikant, die des Hypothalamus und der Medulla zwischen Tag 1 und 20. Zwischen Rassen waren keine statistisch signifikanten Unterschiede in CCK-IR. Unsere Ergebnisse zeigen einen Anstieg in CCK-IR in Gehirnregionen, die bei Futteraufnahme involviert sind, und k?nnten der F?higkeit junger Schweine zur Assimilation von N?hrstoffen aus festen Rationen parallel sein. RéSUMé: Immunoréactivité de type cholécyctokinine dans le cerveau de jeunes porcs Meishan et Duroc La cholécystokinine (CCK) est peptide présent à la fois dans le tractus gastro-intestinal et le cerveau. Son influence sur le contr?le de la consommation alimentaire a été mise en évidence dans plusieurs espèces d'animaux dont le porc. Les races porcines chinoises comme la Meishan présentent une faible croissance et un dép?t de gras important. Cette étude décrit les concentrations régionales des composés présentant une réactivité immunologique similaire à la cholécystokinine (CCK-IR), déterminés par dosage radio-immunologique, dans le cerveau de porcelets Duroc et Meishan. Des cerveaux de porcelets de chacune des deux races ont été examinés à 1, 10 et 20 jours après la naissance. La CCK-IR augmente avec l'age dans les trois zones du cerveau examinées (cortex, moelle et hypothalamus). Les concentrations corticales augmentent significativement etre 1 et 10 jours d'a ainsi qu'entre 10 et 20 jours d'age. Les concentrations dans l'hypothalamus et la moelle croissent significativement entre 1 et 20 jours d'age. Les différences de CCK-IR entre races ne sont statistiquement significatives pour aucun des trois ages étudiés. Nos résultats montrent qu'un accroissement de CCK-IR dans les régions du cerveau impliquées dans le contr?le de la consommation alimentaire peutètre mis en parallèle avec la capacité de porcelets à assimiler les éléments nutritifs d'un régime alimentaire solide. RESUMEN: Immunoreactividad similar a la de la colecistoquinina en los cerebros de lechones jovenes Meishan y Duroc La colecistoquinina (CCK), un péptido que se encuentra en el sistema digestivo y el cerebro, esta rela-cionado con el control del apetito en varias especies, incluido el cerdo. Las razas chinas como la Meishan se distinguen por su crecimiento lento y deposición de tejido adiposo. En este estudio hemos descrito las concentraciones de sustancias immunoló-gicamente similares a la colecistoquinina (CCK-IR) en tres regiones del cerebro de lechones Duroc y Meishan utilizando radioimmunoensayo. Los cerebros de cerdos de 1, 10 y 20 días de edad de cada raza fueron examinados. La CCK-IR aumentó con la edad en las tres áreas examinadas (córtex, médula e hipotálamo). Las concentraciones en el córtex aumentaron significativamente de 1 a 10 y de 10 a 20 dias de edad. Las concentraciones en el hipotálamo y médula aumentaron significativamente entre los días 1 y 20. No hubo diferencias significativas en CCK-IR entre las razas en ninguna de las edades estudiadas. Nuestros resultados indican que un aumento en CCK-IR en las regiones del cerebro que participan en el control del consumo de alimente podrían ir paralelas a la habilidad de los lechones para asimilar nutrientes de una dieta sólida.     

Inhibitory effect and mechanisms of an anthocyanins- and anthocyanidins-rich extract from purple-shoot tea on colorectal carcinoma cell proliferation

One newly bred variety of tea cultivar, purple-shoot tea, was selected to evaluate its antiproliferative effects on colorectal carcinoma cells, as well as normal colon cells. The phytochemicals and identified catechins of purple-shoot tea extract (PTE) were significantly higher than that of ordinary tea, especially the anthocyanins (surpassed by 135-fold) and anthocyanidins (surpassed by 3.5-fold). PTE inhibited the proliferation of COLO 320DM (IC(50) = 64.9 μg/mL) and HT-29 (IC(50) = 55.2 μg/mL) by blocking cell cycle progression during the G(0)/G(1) phase and inducing apoptotic death. Western blotting indicated that PTE induced cell cycle arrest by reducing the expression of cyclin E and cyclin D1 in COLO 320DM and the upregulation of p21 and p27 cyclin-dependent kinase inhibitors in HT-29. Two cells treated with PTE also indicated the cleavage of PARP, activation of caspase 3, and an increased Bax/Bcl-2 ratio. Our results showed that PTE is a potential novel dietary agent for colorectal cancer chemoprevention.     

Protective Effect of Deferricoprogen Isolated from Monascus purpureus NTU 568 on Citrinin-Induced Apoptosis in HEK-293 Cells

Monascus species have traditionally been used in Asian food, with rice as their fermentation substrate. Red mold rice (RMR) contains citrinin, a nephrotoxic agent capable of exerting oxidative stress and cellular apoptosis. We investigated the components in RMR that could minimize the adverse effects of citrinin. Combining chemical separations and bioactivity assays, we identified an antioxidative component called deferricoprogen (DFC) in the fermented rice of Monascus purpureus NTU 568. The DFC structure was confirmed by nuclear magnetic resonance (NMR) and mass spectra analysis. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) free-radical-scavenging activity of DFC was similar to that of vitamin E. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay and flow cytometric analysis showed the effect of DFC and citrinin on cell viability and cell cycle. DFC was found to be protective against the cytotoxicity and cell death induced by citrinin on human embryonic kidney (HEK-293) cells. DFC also demonstrated anti-apoptotic property in preventing citrinin-induced apoptosis.     

Purification and characterization of two chitosanase isoforms from the sheaths of bamboo shoots

Two thermally stable chitosanase isoforms were purified from the sheaths of chitosan-treated bamboo shoots. Isoforms A and B had molecular masses of 24.5 and 16.4 kDa and isoelectric points of 4.30 and 9.22, respectively. Using chitosan as the substrate, both isoforms functioned optimally between pH 3 and 4, and the optimum temperatures for the activities of isoforms A and B were 70 and 60 °C, respectively. The kinetic parameters K(m) and V(max) for isoform A were 0.539 mg/mL and 0.262 μmol/min/mg, respectively, and for isoform B were 0.183 mg/mL and 0.092 μmol/min/mg, respectively. Chitosans were susceptible to degradation by both enzymes and could be converted to low molecular weight chitosans between 28.2 and 11.7 kDa. Furthermore, the most susceptible chitosan substrates were 50-70 and 40-80% deacetylated for isoforms A and B, respectively. Both enzymes could also degrade chitin substrates with lower efficacy. N-Bromosuccinimide and Woodward's reagent K strongly inhibited both enzymes.     

Klymollins T–X,Bioactive Eunicellin-Based Diterpenoids from the Soft Coral Klyxum molle

Five new eunicellin-based diterpenoids, klymollins T–X (1–5), along with two known compounds (6 and 7) have been isolated from the soft coral Klyxum molle. The structures of these new metabolites were elucidated by extensive spectroscopic analysis and by comparison with related known compounds. Compound 5 was found to exert significant in vitro anti-inflammatory activity against LPS-stimulated RAW264.7 macrophage cells. Furthermore, compounds 4 and 7 were shown to exhibit cytotoxicity against a limited panel of human cancer cell lines.     

Effect of Phosphate Salts on the Pasting Properties of Korean Instant‐Fried Noodle

Up to 15% modified potato starch (MPS) is often added into Korean instant noodle formulas to improve the cooking and eating quality. However, the addition of MPS leads to higher raw material costs. The effect of phosphate salts (PS) on Korean instant‐fried noodle pasting properties was investigated in this research. When 33% of total MPS in the regular Korean instant noodle formula was substituted by a small amount of PS (≤0.3%), the results clearly indicated that all the PS improved the pasting properties of noodle dough and ground instant‐fried noodle powder, especially the peak viscosity of the Rapid ViscoAnalyser (RVA) curves. However, the pasting temperature was affected little by PS except that tripotassium phosphate increased it significantly. The RVA results showed that the pasting properties of ground instant noodle powder were significantly improved when 33% of MPS was substituted by 0.300% dipotassium phosphate. Meanwhile, the pasting properties were close to those of the regular formula ground instant noodle powder when 33% of MPS was substituted by 0.030% monosodium phosphate or 0.300% disodium phosphate. Therefore, we suggest that MPS used in Korea instant‐fried noodle could be partially substituted by the blends of selected PS.     

The effect of polyclonal antibody on intracellular calcium increase induced by Mycoplasma hyopneumoniae in porcine tracheal cells

An investigation was undertaken to assess whether polyclonal convalescent and hyperimmune sera obtained from pigs inhibit Mycoplasma hyopneumoniae induced increases in intracellular calcium [Ca2+](i) in ciliated porcine tracheal cells. Basal [Ca2+](i) in the tracheal cells was 97+/-13 nM (n=22 cells in four experiments) and after exposure to M. hyopneumoniae (300 micro g/mL or 10(11) CCU/mL), [Ca2+](i) increased by 246+/-56 nM within 100 s. After pre-treatment with hyperimmune or convalescent serum, M. hyopneumoniae increased [Ca2+](i) by 196+/-43 and 223+/-65 nM, respectively. It was found that neither hyperimmune nor convalescent serum significantly prevented the increase in [Ca2+](i) compared with M. hyopneumoniae alone. It was concluded that polyclonal antibodies produced by mycoplasma vaccination or exposure to the pathogen do not prevent M. hyopneumoniae-induced increase in [Ca2+](i).     

Development of the activities of pancreatic and caecal enzymes in White Roman goslings

1. This study was to investigate the development of the activities of pancreatic and caecal enzymes in White Roman goslings from hatching to 28 d of age. 2. A total of 80 1-d-old goslings were used. At hatching, 3 and 7 d of age, 16 goslings (8 males and 8 females) were used. At 11, 14, 21 and 28 d of age, 8 goslings, 4 males and 4 females were selected. The activities of amylase, lipase, trypsin and chymotrypsin in each segment of the small intestine (duodenum, jejunum, ileum) and their contents and the activity of cellulase in the caecal contents were measured. 3. The specific activity (SA) of amylase in the duodenal mucosa and contents increased significantly both from 11 to 14 and 14 to 21 d of age and declined after 28 d of age. In the jejunum and ileum, there was a significant peak in the SA of amylase in the mucosa in goslings from 7 to 21 d of age. On average, the SA of lipase of mucosa and content in the small intestine was initially low but gradually increased from 14 to 21 and from 21 to 28 d of age. 4. The SA of trypsin in the contents of the duodenum and the jejunum increased both from 7 to 11 and 11 to 14 d of age and there was high activity in the ileal contents from 14 to 21 d of age. From 3 to 14 d of age, the SA of chymotrypsin in the duodenal mucosa significantly increased and peaked. There was a significant peak at 11 d of age in the SA of chymotrypsin in the duodenal or jejunal contents and in the ileal contents at 7 d. The SA of cellulase in the caecal contents increased linearly with age, reaching a plateau at 28 d of age. 5. The rate of activity development varied from hatching to 28 d of age. However, the mean SA of amylase and lipase of intestinal contents reached a peak at 21 d, trypsin and chymotrypsin at 11 d and cellulase at 28 d of age, respectively. The quantitative changes in SA of cellulase, amylase, lipase, trypsin and chymotrypsin increased by about 4-, 3-, 5-, 2- and 3-fold, respectively, in the intestinal content of goslings. Thus, development of proteases in the intestine of goslings peaked more rapidly than amylase, lipase and cellulase during the first 4 weeks.     

Curcuminoids-cellular uptake by human primary colon cancer cells as quantitated by a sensitive HPLC assay and its relation with the inhibition of proliferation and apoptosis

Curcumin, which is a bright orange-yellow pigment of turmeric with antioxidant properties, has been shown to produce a potent preventative action against several types of cancers in recent studies. It has also been reported to protect the development of colon tumor in animals being fed with carcinogen. In the colon cancer cells, curcumin was illustrated to inhibit cell proliferation and induce apoptosis. As an antioxidant, it acts as an anti-inflammatory as well as an antitumor agent. Curcumin has been detected to exist in nature in the form of curcuminoids, a mixture of curcumin, the major component, with two of its related demethoxy compounds (demethoxycurcumin and bisdemethoxycurcumin). In the present study, we have investigated the antiproliferation and induced apoptosis effects of curcuminoids on colon cancer, using the primary cancer cells isolated from Taiwanese colon cancer patients as the model for colorectal cancer. Results showed that curcuminoids inhibited cell proliferation and induced apoptosis of these human primary colon cancer cells. The effects were observed in a dose-dependent manner as dose increased from 12.5 to 100 microM. With the aim of furthering the fundamental understanding of the mechanisms underlying the antiproliferation and induced apoptosis effects of curcuminoids on these human colon cancer cells, we developed a sensitive, rapid, and reproducible assay method based on high-performance liquid chromatography (HPLC). This HPLC technique developed was found to successfully determine, in a quantitative manner, the cellular uptake of curcuminoids. The uptake of these curcuminoids by the colon cancer cells was shown to increase as the dose of curcuminoids was increased. The observations of inhibited proliferation and increased apoptosis in the colon cancer cells appeared to be associated with the cellular uptake of curcuminoids.     

Proteomic analysis of pycnogenol effects in RAW 264.7 macrophage reveals induction of cathepsin D expression and enhancement of phagocytosis

Pycnogenol, polyphenolic compounds extracted from the pine bark, is beneficial for human health. To understand more of its effects, the present study is to explore the protein expression pattern induced by pycnogenol in RAW 264.7 cells. Global analysis using two-dimensional gel electrophoresis indicated that treatment with pycnogenol induces upregulation of four proteins, whose identities were revealed by mass spectrometry as cathepsin D, keratinocyte lipid-binding protein, proteasome subunit alpha type 1, and annexin IV. The pycnogenol effect displayed a time- and concentration-dependent manner. Unlike pycnogenol, N-acetyl cysteine and vitamin C had no effect on cathepsin D expression. Further studies showed that cathepsin D induction is correlated with an increase of lysosomal staining and enhancement of phagocytosis. These results reveal the novel effects of pycnogenol on protein expression and phagocytic functions and illustrate the advantage of proteomics-based strategy in unveiling the molecular basis of phytochemicals.