Differential effects of flavonoids on barrier integrity in human intestinal Caco-2 cells

Flavonoids, present in fruits, vegetables, and teas, provide beneficial effects for our health. We investigated the effect of a number of flavonoids on tight junction (TJ) barrier integrity in human intestinal Caco-2 cells. Transepithelial electrical resistance (TER; a TJ integrity marker) across cell monolayers was measured in cells incubated with flavonoids for 24 h. Chrysin decreased the TER, indicating a decrease in TJ integrity. Daidzein, hesperetin, naringenin, and morin increased the TER, indicating increased TJ integrity. Luteolin and genistein increased or normalized the TER after a transient decrease. Immunoblot analysis revealed that these changes in TER were caused by modification of the cytoskeletal association and expression of TJ proteins, zonula occludens (ZO)-1, ZO-2, occludin, junctional adhesion molecule-1, and/or claudins. Our results suggest that various flavonoids participate in the regulation of intestinal TJ barrier integrity and that this regulation may partially contribute to the flavonoid-mediated biological effects on our health.     

Kinematic gait analysis and lactation performance in dairy cows fed a diet supplemented with zinc,manganese, copper and cobalt

This study investigated how supplementation of the diet of dairy cows with trace minerals (zinc, manganese, copper and cobalt) affected kinematic gait parameters and lactation performance. Eight Holstein cows were divided into two groups, with each group receiving a different dietary treatment (control diet, or control diet supplemented with trace minerals) in a two‐period crossover design. Kinematic gait parameters were calculated by using image analysis software. Compared to cows fed the control diet, cows that received the trace mineral‐supplemented diet exhibited significantly increased walking and stepping rates, and had a shorter stance duration. Feed intake and milk production increased in cows fed the trace mineral‐supplemented diet compared with control groups. The plasma manganese concentration was not different in control and experimental cows. In contrast, cobalt was only detected in the plasma of cows fed the supplemented diet. These results provide the first evidence that trace mineral supplementation of the diet of dairy cows affects locomotion, and that the associated gait changes can be detected by using kinematic gait analysis. Moreover, trace mineral supplementation improved milk production and only minimally altered blood and physiological parameters in dairy cows.     

Comparison of learning ability and memory retention in altricial (Bengalese finch,Lonchura striata var. domestica) and precocial (blue‐breasted quail,Coturnix chinensis) birds using a color discrimination task

The present study sought to assess the potential application of avian models with different developmental modes to studies on cognition and neuroscience. Six altricial Bengalese finches (Lonchura striata var. domestica), and eight precocial blue‐breasted quails (Coturnix chinensis) were presented with color discrimination tasks to compare their respective faculties for learning and memory retention within the context of the two developmental modes. Tasks consisted of presenting birds with discriminative cues in the form of colored feeder lids, and birds were considered to have learned a task when 80% of their attempts at selecting the correctly colored lid in two consecutive blocks of 10 trials were successful. All of the finches successfully performed the required experimental tasks, whereas only half of the quails were able to execute the same tasks. In the learning test, finches required significantly fewer trials than quails to learn the task (finches: 13.5 ± 9.14 trials, quails: 45.8 ± 4.35 trials, P < 0.05), with finches scoring significantly more correct responses than quails (finches: 98.3 ± 4.08%, quails: 85.0 ± 5.77% at the peak of the learning curve). In the memory retention tests, which were conducted 45 days after the learning test, finches retained the ability to discriminate between colors correctly (95.0 ± 4.47%), whereas quails did not retain any memory of the experimental procedure and so could not be tested. These results suggested that altricial and precocial birds both possess the faculty for learning and retaining discrimination‐type tasks, but that altricial birds perform better than precocial birds in both faculties. The present findings imply that developmental mode is an important consideration for assessing the suitability of bird species for particular experiments.     

Genogrouping of birnaviruses isolated from marine fish: a comparison of VP2/NS junction regions on genome segment A

Yellowtail ascites virus and related strains isolated from marine fish have been shown to be similar to infectious necrosis virus (IPNV) in terms of biological and serological characteristics. This paper explores the relationship of aquatic birnaviruses at the genetic level. The junction region on the genome segment A coding viral capsid protein VP2 and viral protease NS was amplified by PCR in six marine strains. Analysis of nucleotide and the deduced amino acid sequences revealed that the six marine strains have amino acid variations in the possible amino terminus of NS when compared to IPNV. The six marine strains form a new genogroup which is distinguished from three serotypes ofPNV.     

Identification of a heparin-binding,mesoderm-inducing peptide in the swim-bladder of the red seabream,Pagrus major: a probable fish fibroblast growth factor

The brain, pituitary, liver, ovary and swim-bladder of the red seabream (Pagrus major) were screened for the presence of a growth factor capable of stimulating the proliferation of BHK-21 cells, and the swimbladder extract was found to exert the most powerful activity. Swim-bladder derived growth factor (SBDGF) with heparin-binding ability was isolated by heparin-affinity and size-exclusion chromatography using heparin-Sepharose CL-6B and Superose 12, respectively. The relative molecular mass of SBDGF was 22,500 and the isoelectric point was 9.4. The growth of mammalian fibroblasts (BALB/3c 3T3, BHK-21) and endothelial cells (CPAE) was markedly stimulated by SBDGF. Its growth-promoting activity was also effective in fish fibroblasts (BF-2 and RTG-2), and was heat- and acid-labile. In the mesoderm-induction assay, explants of presumptive ectoderm dissected from Xenopus laevis blastulae (stage 9) were incubated with SBDGF. SBDGF induced ventral mesoderm in the explants, including hemocyte-like cells, mesenchyme and coelomic epithelium. These properties suggest that SBDGF is a fish fibroblast growth factor and that the swim-bladder is the source.     

Ultrastructural characteristics of two distinct gonadotropes (GTH I- and GTH II-cells) in the pituitary of rainbow trout Oncorhynchus mykiss

The salmonid pituitary produces two chemically distinct gonadotropins (GTHI and GTHII). Ultrastructural characteristics of GTHI- and GTHII-producing cells were studied in the trout pituitary with electronmicroscopic immunocytochemistry using antisera against salmon GTHIβ- and IIβ-subunits. In pituitaries from vitellogenic fish, GTHI-cells were characterized by numerous dilated cisternae of the granular endoplasmic reticulum (GER) and a small number of Iβ-positive granules (diameter, 100–300 nm), whereas GTHIIβ-immunoreactivity was found on granules (diameter, 200–400 nm) and large globules (diameter, 500–4000 nm) in apparently different cells (GTHII-cells). Distinct cellular distributions of GTHI and GTHII were maintained during gametogenesis, although morphological characteristics of GTHI- and GTHII-cells overlapped each other due to changes in number and size of the granules, globules and cisternae of the GER. Interestingly, the globules in the GTHI-cells were immunonegative for GTHIβ, although in the GTHII-cells they were always stained with GTHIIβ-antiserum. These results confirm that GTHIβ and GTHIIβ are synthesized in distinctly different cell-types in the salmonid pituitary and indicate that morphological characteristics cannot be used to distinguish these two cell-types.

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Résumé L'hypophyse des salmonidés produit deux gonadotropines (GTHI et GTHII) chimiquement distinctes. Les caractéristiques ultrastructurales des cellules produisant la GTHI et la GTHII ont été étudiées par immunocytochimie en microscopie électronique en utilisant des anticorps dirigés contre les sousunités GTHIβ et GTHIIβ de saumon. Chez les poissons en vitellogenèse, les cellules à GTHI se caractèrisent par un réticulum endoplasmique granulaire (REG) contenant de nombreuses citernes dilatées et un petit nombre de granules positives à la GTHIβ (diamètre 100–300 nm) tandis qu'une immunoréactivité à la GTHII était trouvée sur des granules (diamètre 200–4000 nm) et de grands globules (diamètre 500–4000 nm) dans des cellules apparemment différentes (cellules à GTHII). Des distributions cellulaires distinctes de la GTHI et la GTHII se sont maintenues à des stades plus avancées de la gamétogenèse, bien que les caractéristiques morphologiques des cellules à GTHI et GTHII se recoupent suite à des changements dans le nombre et la taiile des granules, des globules et des citernes du REG. II est à remarquer que les globules prśents dans les cellules à GTHI étaient immunonégatifs à la GTHIβ alors que dans les cellules à GTHII ils étaient toujours marqués par l'anticorps contre la GTHIIβ. Ces résultats confirment que, dans l'hypophyse des salmonidés, les GTHIβ et GTHIIβ sont synthetisées dans des types cellulaires différents et indiquent que des caractéristiques morphologiques ne peuvent être utilisées pour distinguer ces deux types cellulaires.

     

Effects of temperature on vitellogenesis in Japanese eel Anguilla japonica

ABSTRACT:   Cultured immature female Japanese eels acclimated to sea water at either 10 or 20°C were treated weekly with salmon gonadotropin fraction (sGTH) in order to investigate the effects of water temperature on artificial induction of ovarian maturation. In eels maintained at 20°C, ovulation was induced in 11 of 18 fish during the experimental period of 13 weeks, whereas at 10°C all fish showed a low gonadosomatic index (GSI) at the end of the experiment. Plasma vitellogenin levels were higher in eels kept at 20°C than in eels at 10°C throughout the experiment. However, no significant differences were observed in the plasma testosterone and estradiol-17β levels between groups. Eels pretreated with sGTHs at 10°C for 13 weeks were separated to two different temperature (10 and 20°C) groups, and received the same weekly sGTH injections. In eels transferred to 20°C, plasma vitellogenin levels, GSI and oocyte diameter were increased, but these values were maintained at low levels in eels that remained at 10°C. These results clearly indicate that water temperature is an important factor regulating vitellogenesis in the Japanese eel.     

DNA markers linked to Pga1, an adzuki bean gene that confers resistance to Cadophora gregata race 1

Brown stem rot (BSR) caused by Cadophora gregata f. sp. adzukicola (syn. Phialophora gregata) is a serious soilborne disease of adzuki bean (Vigna angularis) in Japan. Cultivation of resistant cultivars is the most effective disease control method, therefore the selection of resistant lines is a priority for breeders. BSR-resistant adzuki bean lines have been screened in pathogen-infected fields. However, field selection using the pathogen and artificial inoculation methods is time-consuming and labor-intensive. In the present study, we used 105 F₃ lines derived from a cross between a BSR-resistant cultivar ‘Syumari’ and a susceptible cultivar ‘Buchishoryukei-1’ for BSR inoculation tests. Amplified fragment-length polymorphism (AFLP) analyses with 1024 primer sets revealed that six fragments were polymorphic between resistance and susceptible bulked groups. Five DNA markers (Pg77, Pg118, Pg138, Pg139 and Pg126) were developed from the nucleotide sequences of polymorphic AFLP markers and their flanking regions. Pg118, which was derived from E-ACT/M-ACT-118, was tightly linked to the resistance gene Pga1 and was converted into a codominant marker for its easier use in marker-assisted selection for adzuki bean BSR resistance. Finally, the applicability of the developed markers for BSR resistance was tested on 32 adzuki bean accessions or cultivars.