Human interleukin-1β induces A375-S2 human melanoma apoptosis through caspase pathway

AIM: To study the molecular biological mechanism and signal transduction pathway of interleukin-1β (IL-1β)-induced apoptosis in A375-S2 melanoma cells. METHODS: Photomicrocropy showed typical apoptotic changes. The cytotoxic effect of IL-1β in vitro and influences of caspases in this effect were measured by MTT assay. The cytotoxicity of cells was assessed by LDH-based assay. Degradation of DNA was detected by agarose gel electrophoresis. RESULTS: The inhibitory effect of IL-1β on A375-S2 cell growth was in a dose and time-dependent manner, and cell death rate reached more than 90% at 72 h after treatment with 10^-9mol/L IL-1β. The inhibitors of caspase-family, -1, -3, -8, -9, and -10, partially blocked cell death at early stage. LDH assay showed that major IL-1β-induced cell death was apoptosis, and in a dose and time-dependent manner. Typical apoptotic DNA ladder was observed in agarose gel electrophoresis. CONCLUSION: IL-1β induced apoptosis in melanoma A375-S2 cells by activating caspase pathway.     

Clinical evaluation of growth hormone secretion in cattle using insulin tolerance test

Growth hormone secretion was evaluated in cattle. Clinically healthy bovine growth hormone (bGH) concentrations were 10.7 +/- 1.6 ng/ml in Holstein and 7.8 +/- 3.9 ng/ml in Japanese black cattle. The bGH concentration alternated at three-hour intervals, and tended to be higher at midnight and lower in the morning and before feeding. Insulin tolerance test (ITT) at an insulin dosage of 0.25 U/kg showed a significant increase of bGH concentration to 331 +/- 153% at 60 to 90 min after injection. In ITT applied to five under-growth calves of Japanese black cattle, the basal bGH concentrations were lower and peak values after insulin injection were shown to be significantly low. The ITT is useful for the clinical examination of bGH secretion in cattle.     

Host selection of small cedar longicorn beetle, <Emphasis Type="Italic">Callidiellum rufipenne</Emphasis> (Coleoptera:Cerambycidae), on Japanese cedar, <Emphasis Type="Italic">Cryptomeria japonica</Emphasis>, in terms of bark water content of host trees

We caught the adults of a secondary wood-boring insect, Callidiellum rufipenne (Motschulsky), on Japanese cedar, Cryptomeria japonica D. Don., trees and logs with manipulated bark water contents to clarify the mechanism of host selection by C. rufipenne. More C. rufipenne were trapped when the bark water content range was ca. 120–200%. Fewer C. rufipenne were trapped at higher and lower bark water contents. These findings indicate that C. rufipenne adults do not simply select weakened or dying trees. Bark water content is one of important factors for C. rufipenne choosing their host for oviposition.     

Changes in estrogen receptor expression in the chick thymus during late embryonic development

In chickens, although estrogen receptors (ER) are reported to be associated with the immunological processes, detailed information about the differences in ER expression in the tissues related to the development of lymphocytes is not fully known, especially during the developmental stage. To learn more about this immunological relationship, we used semi‐quantitative polymerase chain reaction method to detect the ER expression levels in the thymus tissues of chicks during the developmental stage. Furthermore, ER‐expressing cells were detected by immunohistochemistry. The results of this study show that the expression level of ER increased on embryonic day 16 and decreased on day 20. Furthermore, ER expression was significantly higher in male than in female chickens at day 16. The increased expression on day 16 and decreased level on day 20 were also reproduced in the incidence of immunoreactive cells, although there was a 1‐day delay in the elevated incidence of the cells. This study revealed the changes in ER expression and the incidence of ER‐positive cells in the thymus of chickens during the developmental stage.     

Distribution of three GnRHs in the brain and pituitary of the wild Japanese flounder Paralichthys olivaceus

ABSTRACT:   Wild adult maturing and immature female Japanese flounder Paralichthys olivaceus were collected in June 2004 and January 2005, respectively, to clarify a possible role of gonadotropin-releasing hormones (GnRHs) in reproduction. Levels of salmon GnRH (sGnRH), chicken GnRH-II (cGnRH-II) and sea bream GnRH (sbGnRH) in the brain and pituitary were examined by time-resolved fluoroimmunoassay. Three forms of GnRHs were detected in the discrete brain at various levels. In the pituitary of both maturing and immature fish, sbGnRH was abundant together with a pronounced amount of sGnRH, whereas cGnRH-II was almost below the detectable limit. In maturing fish, levels of sbGnRH were high in the telencephalon, hypothalamus and pituitary, while levels of sbGnRH of immature fish were very low in these regions. These results indicate that sbGnRH is mainly responsible for gonadotropin secretion, and that sbGnRH in the anterior part of the brain is associated with gonadal maturation in the Japanese flounder.     

Effects of photoperiod on gonadotropin-releasing hormone levels in the brain and pituitary of underyearling male barfin flounder

ABSTRACT:   A pleuronectiform fish, the barfin flounder Verasper moseri , expresses three gonadotropin-releasing hormone (GnRH) forms in the brain: salmon GnRH (sGnRH), chicken GnRH-II (cGnRH-II) and seabream GnRH (sbGnRH). To clarify the effects of photoperiod on GnRH systems, changes in brain and pituitary GnRH peptide levels were examined using time-resolved fluoroimmunoassays. In experiment 1, 5-month-old male barfin flounder (mean total length 9.0 cm, body weight 11.0 g) were divided into short (8:16 h light : dark [L:D] cycle; lights on 08.00–16.00 hours) and long photoperiod (16:8 h L:D cycle; lights on 04.00–20.00 hours) groups in mid September and maintained until November under natural water temperature (19.3–15.2°C). Brain sGnRH concentrations were significantly higher in the 16:8 h L:D group than in the 8:16 h L:D group, whereas no significant differences were observed in total length, body weight, plasma testosterone concentration, brain cGnRH-II concentration and pituitary sbGnRH content. In experiment 2, 7-month-old male barfin flounder (mean total length 16.5 cm, body weight 76.8 g) were divided into short and long photoperiod groups in mid December and maintained until February under natural water temperature (12.5–6.6°C). Total length, body weight and condition factor were significantly greater in the 16:8 h L:D group than in the 8:16 h L:D group, whereas no significant differences were observed in plasma testosterone concentration and GnRH levels in the brain and pituitary. These results indicate that levels of sGnRH in barfin flounder are influenced by photoperiodic treatment dependent on water temperature and/or body size.     

Production of aromatic amino acids and related compounds from p-hydroxyphenylacetic acid by rumen microorganisms in vitro

Suspensions of mixed rumen bacteria (B), protozoa (P), and mixed rumen microorganisms (BP) prepared from rumen contents of fistulated goats were anaerobically incubated with 1 mM p‐hydroxyphenylacetic acid (HPA) at 39°C for 24 h. Tyrosine (Tyr), phenylalanine (Phe), tryptophan (Trp) and other related compounds in both supernatants and hydrolyzates of microbial cells in all incubations were analyzed by HPLC. Large amounts of Tyr (32.1, 42.7 and 36.1% of disappeared HPA in B, P and BP, respectively) were produced from HPA during a 12 h incubation period. The formation of Tyr in P (178.6 µmol/g MN) was 1.5 and 2 times higher than in B and BP, respectively. Phe (7–11% of the disappeared HPA) and Trp (3–6% of the disappeared HPA) were also synthesized from HPA in B, P, and BP. Phe synthesis in P (46.3 µmol/g MN) was 1.7 times higher than in B but, in contrast, Trp synthesis in B, was 1.6 times higher than in P. The metabolites p‐hydroxyphenylpyruvic acid (in the range of 5–14% of disappeared HPA), phenylacetic acid (1–11%), p‐hydroxybenzoic acid (3–7%) and benzoic acid (1–6%) were produced from HPA in B, P and BP. Phenylpropionic acid (6% of the disappeared HPA) was produced only in B and BP.     

Effect of dietary addition of seaweed and licorice on the immune performance of pigs

In pig production, dietary additive antibiotics are usually used for growth stimulation and disease prevention, although there is public concern about the increased incidence of resistant antibiotics and food safety. It is possible that such antibiotics might be replaced by naturally derived products such as seaweed and licorice. In this study, we evaluated the effect of dietary addition of seaweed and licorice on enhancing the immune function in swine. The animals of each group (eight animals per group) were sensitized at day 42 and 49, and the immunoglobulin production and the expression of cytokines were detected by the ELISA and real‐time PCR. As the results, saliva IgA production of the seaweed‐treated group increased around five times compared to that of control (day 56). Delayed hypersensitivity reaction and IgG production of the seaweed‐treated group increased around 1.8–2.0 times. In addition, enhanced saliva IgA production was detected at day 50 (around two times) and day 51 (around five times) by the licorice treatment, and lower expression level of tumor necrosis factor‐α messenger RNA at day 51 (around 1/25) was observed in the licorice treatment. We conclude that the replacement of antibiotics by naturally derived dietary additives might be feasible for immune system enhancement.