Effects of leptin and tumor necrosis factor-alpha on degranulation and superoxide production of polymorphonuclear neutrophils from Holstein cows

Leptin, a pleiotropic hormone regulating food intake and energy expenditure, has been shown to directly modulate human polymorphonuclear neutrophil (PMN) functions or indirectly through the action of tumor necrosis factor-alpha (TNF-alpha). Bovine PMN have considerable different characteristics from human PMN. For example, it does not respond to N-formyl-Methionyl-Leucyl-phenylalanine, a well known human PMN activator. In the present study, we tested the effects of leptin and TNF-alpha on superoxide production and degranulation of bovine peripheral PMN, in which both long isoform of leptin receptor (Ob-Rb) and TNF receptor 1 were expressed. Human leptin, human TNF-alpha, phorbol myristate acetate (PMA) and opsonized zymosan particles (OZP) did not stimulate degranulation responses, while zymosan-activated serum (ZAS) did. Neither leptin nor TNF-alpha enhanced the ZAS-induced degranulation responses. TNF-alpha, PMA, OZP and ZAS increased superoxide production in different magnitudes, whereas leptin did not. TNF-alpha, but not leptin, enhanced OZP- and ZAS-induced superoxide production, possibly, in part due to facilitating translocation of p47(phox), a component of NADPH oxidase. These results indicate that, unlike in human PMN, leptin does not have any direct effect on degranulation and superoxide production in bovine PMN, although TNF-alpha influences superoxide production.     

Effect of ovary lipid of skipjack tuna on rat serum components after stress application

Lipid extracted from the ovary of skipjack tuna by the method that we developed is rich in phospholipid-type docosahexaenoic acid. The ovary lipid of skipjack tuna (OLS) was studied for its anti-stress activity in male Wistar rats, focusing on stress-related blood components: recovery from stress was examined after application of water immersion restraint stress. As a result, serum corticosterone (CORT) secretion was inhibited and decreased rapidly after stress application in rats given OLS compared with control rats. As CORT acts as a glucocorticoid, non-esterified fatty acid (NEFA) is expected to increase by stress application. However, the concentration tended to be lower in rats given OLS than in control rats. With respect to OLS concentration, OLS increased serum dehydroepiandrosterone, secretion concentration-dependently. In addition, as with the recovery study, it tended to inhibit the increase in NEFA. These results indicate that OLS may have an anti-stress activity against acute stress.     

Influence of light intensity on feeding, growth, and early survival of leopard coral grouper (Plectropomus leopardus) larvae under mass-scale rearing conditions

This study investigated the effect of different light intensities on feeding, growth and survival of early stage leopard coral grouper Plectropomus leopardus larvae. Four different light intensities (0, 500, 1000 and 3000 lx) were used and larvae were kept under constant light conditions from 0 day after hatching (DAH) to 5 DAH. The larvae were fed a small S-type of Thai strain rotifers at a density of 20 individuals/mL from 2 DAH. The number of rotifers in larval digestive organ and total length of larvae were examined at 3 h intervals between 04:00 and 22:00 h on 3 DAH, and thereafter at 6 h intervals until the end of the experiment (5 DAH). Four experimental trials of the larval rearing were repeated using by 60 kL mass-scale rearing tanks. The results indicate that coral grouper larvae are visual feeders and their food intake increases with increasing light intensity. Food intake of larvae reared at 3000 lx was significantly higher than those reared at 0–1000 lx on 3 DAH despite being the first-feeding day (P < 0.01). On 4 DAH, total length of larvae reared at 3000 lx was significantly larger than those reared at the lower light intensities (0, 500 and 1000 lx), and thereafter light intensity significantly influenced larval feeding and growth until the end of the experiment. Survival on 5 DAH did not show a significant difference between light intensities, but survival rate at 3000 lx and 1000 lx had a tendency to be higher than those reared at the lower light intensities (0 and 500 lx). In contrast, larvae reared at 0 lx exhibited stagnant and/or negative growth. These results indicate that light intensity is significantly the factor affecting larval feeding, growth, and survival in coral grouper larvae under the rearing conditions.     

Surface-active properties of particle size fractions in two humic acids

The relationships between surface active properties and humic acid (HA) particle sizes were investigated. Two HAs from an Ando soil and a Brown forest soil were separated into 6 particle size fractions by gel permeation chromatography. Surface-active properties characterized by surface excess value (\gT mol cm^-2), cross-sectional surface area per molecule (A nm²), critical micelle concentration (CMC g L^-1), efficiency and effectiveness of water surface tension reduction were obtained by the measurement of the surface tension of HA solutions from different particle size fractions. For the HA from the Ando soil, except for the smallest particle size fraction, increasing particle size enhanced the efficiency of reduction of the water surface tension and decreased the CMC, while the effectiveness of reduction of the water surface tension was about the same. The surface activity of the HA from the Ando soil increased with increasing particle size. This phenomenon was similar to the surface activity of a homologous series of surfactants, which increased with increasing alkyl chain length. For the HA from the Brown forest soil, the smallest particle size fraction and three large fractions showed a high efficiency, namely a high surface activity. The smallest fraction from the Brown forest soil showed the highest efficiency and the lowest CMC value. In both HAs, the smallest particle size fraction showed exceptional surface-active properties compared with the other fractions and three fractions with large particle size showed a higher surface activity than other smaller fractions.     

Isolation and culture of rabbit primordial germ cells

Primordial germ cells (PGCs) are embryonic precursors of the gametes of adult animals and are considered stem cells of the germline. Since their proliferation in vitro correlates well with the schedule of developmental changes in vivo, they might be interesting research tools for genomic imprinting, germ-cell tumors and fertility. Furthermore, once primordial germ cells are separated and placed on a feeder layer with cytokines, they become cultured pluripotent cell lines called embryonic germ (EG) cells. EG cells share several important characteristics with embryonic stem (ES) cells as they can also contribute to the germ line of chimeras. To investigate the characteristics of PGCs and establish rabbit EG (rEG) cells, we cultured rabbit PGCs (rPGCs) in vitro with various combinations of leukemia inhibitory factor (LIF), basic fibroblast growth factor (bFGF) and forskolin on inactivated mouse embryonic fibroblast (MEF) feeder layers. The present study found PGC proliferation in early cultures and induction of rEG-like colonies. These cells expressed pluripotent markers, such as alkaline phosphatase activity, OCT-4, Sox-2 and SSEA-1, in the undifferentiated state; however, the cells did not develop into a teratoma when injected into the kidney capsules of SCID mice, although the restricted differentiation potentials to neural cells were determined via embryoid body formation. From these characteristics and further characterization of the germ stem cell markers Vasa, SCP-1 and SCP-3, we suggested that these were hybrid cells with characteristics somewhere between PGC and EG cells.     

Severe nonpurulent encephalitis with mortality and feather lesions in call ducks (Anas platyrhyncha var. domestica) inoculated intravenously with H5N1 highly pathogenic avian influenza virus

One-day-old, 2-wk-old, and 4-wk-old call ducks (Anas platyrhyncha var. domestica) inoculated intravenously with the H5N1 highly pathogenic avian influenza virus A/chicken/Yamaguchi/7/2004 isolate (Ck/Yama/7/04) were examined clinically, pathologically, and virologically. Clinically, the birds exhibited mild-to-severe neurologic signs and corneal opacity. All birds in the 1-day-old group and one bird in the 4-wk-old group died within 4 days after the virus inoculation. Histologic changes were characterized by severe nonpurulent encephalitis and necrotic lesions of feather epithelium on day 3 postinoculation (PI) or later. Focal necrosis of myocardial cells, pancreatic acinar cells, skeletal myocytes, and corneal epithelial cells was observed. Viral antigens were detected in association with necrotic changes. Viruses were isolated from all examined organs including the skin with many feathers. Serum antibody against the virus was detected in all surviving birds on day 10 PI by hemagglutination-inhibition tests. These results suggest that Ck/Yama/7/04 has a pathogenicity that causes neurologic sign, nonpurulent encephalitis with mortality, and feather lesions for call ducks. Feather lesions with viral antigens and the virus isolation from the skin suggest that Ck/Yama/ 7/04 has a predilection for feathers in call ducks.     

Immunohistochemical study on the delayed progression of epithelial apoptosis in follicle-associated epithelium of rat Peyer's patch

It is well known that some caspases in apoptosis is involved in determinant of terminal differentiation and maturation of various cells. Our previous study ultrastructurally clarified the differentiation into M cells from immature microvillous epithelial cells and the redifferentiation from M cells to microvillous epithelial cells in the follicle-associated epithelium (FAE) of rat Peyer's patch. In this study, the difference of epithelial apoptosis between the FAE of Peyer's patch and intestinal villi was immunohistochemically investigated in rat jejunoileum. As a result, cleaved caspase-3 was limited to several epithelial cells at the tip of FAE, whereas almost all of the epithelial cells were cleaved caspase-3 positive in intestinal villi. Cleaved caspase-9 was detected only in a few exfoliating or exfoliated epithelial cells of both FAE and intestinal villi. Nuclear DNA-fragmentation was detected only in several epithelial cells of the tip of FAE, while it was expressed from the middle regions in the intestinal villi. The DNase I expression of the epithelial cytoplasm was much weaker in FAE than in intestinal villi. Bcl-x expression was restricted in the apical cytoplasms of epithelial cells in the FAE, whereas it was restricted in whole cytoplasms in villous epithelial cells. These findings suggest that the progression of the apoptotic process in the epithelial cells of FAE is later than in the intestinal villi, so that the possibility of epithelial differentiation might be remained in the FAE, unlike in the intestinal villi.     

Effects of testosterone on rat placental development

We investigated the morphological effects of testosterone on placental development in a rat model of polycystic ovarian syndrome (PCOS). Testosterone propionate (TP), which was subcutaneously administered to pregnant rats with 5 mg/animal from gestation day (GD) 14 to GD 18, induced a maternal weight reduction without mortality or clinical signs from GD 19 onwards. A decrease in fetal and placental weight, an increase in intrauterine growth retardation (IUGR) rates, and histological changes in the placenta were observed on GD 21 but not on GD15 or 17. Histopathologically, on GD 21, the trophoblast septa thickened, and the maternal sinusoids were narrowed in the labyrinth zone, resulting in a small placenta. Additionally, the placental weight, thickness, and histological morphology in the labyrinth zone on GD 21 in the TP-treated group were nearly identical to those on GD 17 in the control and TP-treated groups. Therefore, it was assumed that the testosterone-induced small placenta was induced in association with the developmental inhibition of the fetal part of the placentas from GD 17 onwards.     

Recording cerebellar neuron activities in swimming goldfish

ABSTRACT:   Neuronal activities were investigated in the cerebellum of immobilized and swimming goldfish Carassius auratus . Extracellularly recorded neural activities of the cerebellum in immobilized goldfish were characterized and classified into five types. Based on the waveforms and recording depths, these five neural activity types were estimated to originate from three identified classes of cerebellar neurons: Purkinje cells, eurydendroid cells, and granule cells. Chronic recording of cerebellar neuron activities in unrestrained goldfish was performed for more than 100 h. During the chronic recordings, a submersible amplifier attached to the goldfish head, and a multielectrode array developed for the present study were used. Neuronal activities in the cerebellum of free-swimming fish could also be classified into five types as in the immobilized condition. Firing patterns of two neurons identified as Purkinje cells and eurydendroid cells were analyzed during turning movements of the goldfish. The firing patterns of these neurons changed in relation to turning movements. Although some improvements are required, the chronic recording method developed in the present study can be applied to further investigations concerning the direct relationship between brain neural activities and certain behavior.