Effects on the Local Immunity in the Testis by Exposure to Di-(2-ethylhexyl) Phthalate (DEHP) in Mice

Exposure to di-(2-ethylhexyl) phthalate (DEHP) has been reported to induce spermatogenic disturbance through oxidant stress and affect the immune system as an adjuvant. However, the effect of DEHP on the testicular immune microenvironment has not yet been investigated. In the present study, we examined the testicular immune microenvironment after exposure to doses of DEHP, previously identified as no-observed-adverse-effect levels. Adult male mice were administered food containing 0%, 0.01% or 0.1% DEHP and then testes were analyzed. The results showed that a slight but significant spermatogenic disturbance appeared in the 0.1% DEHP group but not in the 0.01% DEHP group at 8 weeks. It was also demonstrated that lymphocytes and F4/80- and MHC class II- positive cells were significantly increased with the elevation of IL-10 and IFN-γ mRNA expressions in the testes of not only the 0.1% DEHP group but also the 0.01% DEHP group at 8 weeks. Histochemical analyses involving horseradish peroxidase (HRP) as a tracer showed that a little blood-borne HRP had infiltrated into the lumen of a few seminiferous tubules beyond the blood-testis-barrier in both the 0.1% and 0.01% DEHP groups at 8 weeks. This indicates that a dose of DEHP that has little effects on spermatogenesis can change the testicular immune microenvironment with functional damage of the blood-testis barrier.     

CD56 is expressed exclusively on CD3+ T lymphocytes in canine peripheral blood

CD56+ cells in canine blood leukocytes were characterized by flow-cytometric analysis of peripheral blood of 30 healthy adult beagle-dogs (15 males and 15 non-pregnant females). In 19 of the 30 dogs, anti human CD56 antibody, Leu-19, reacted with 8.8-21.7% of peripheral blood lymphocytes. All CD56+ cells simultaneously expressed CD3 molecules on their surface. Further phenotypic analysis revealed that 50.6+/-13.1% of the CD56+ cells showed CD4-CD8+ phenotype and 43.7+/-10.1% showed CD4+CD8- phenotype. Expression intensity of CD56 on the CD4-CD8+CD56+ cells was significantly higher than that on CD4+CD8-CD56+ cells (P<0.001). These findings indicate that CD56, which is a neural cell adhesion molecule, is uniquely expressed on subsets of T lymphocytes in canine peripheral blood.     

A low pathogenic H5N2 influenza virus isolated in Taiwan acquired high pathogenicity by consecutive passages in chickens

H5N2 viruses were isolated from cloacal swab samples of apparently healthy chickens in Taiwan in 2003 and 2008 during surveillance of avian influenza. Each of the viruses was eradicated by stamping out. The official diagnosis report indicated that the Intravenous Pathogenicity Indexes (IVPIs) of the isolates were 0.00 and 0.89, respectively, indicating that these were low pathogenic strains, although the hemagglutinin of the strain isolated in 2008 (Taiwan08) had multibasic amino acid residues at the cleavage site (PQRKKR/G). In the present study, these H5N2 viruses were assessed for their intravenous and intranasal pathogenicity for chickens. It was examined whether Taiwan08 acquires pathogenicity through consecutive passages in chickens. Intravenous pathogenicity of Taiwan08 depended upon the age of the chickens used for the IVPI test; all of the eight-week-old chickens intravenously inoculated with Taiwan08 showed clinical signs but survived for ten days post inoculation (IVPI=0.68), whereas all the six-week-old chickens died (IVPI=1.86). Taiwan08-P8, which were passaged in chickens for eight times, killed all the eight-week-old chickens (IVPI=2.36). The four-week-old chickens died after intranasal inoculation of Taiwan08-P8, indicating that Taiwan08 must have become highly pathogenic during circulation in chicken flocks. These results emphasize the importance of a stamping out policy for avian influenza even if the IVPI of the causal virus is low.     

Effects of methoxychlor exposure during perinatal period on reproductive function after maturation in rats

Methoxychlor (MXC) is a non-steroidal pesticide that is known to possess estrogenic activity, and therefore may be potentially hazardous to the development and/or reproduction. The present study assessed the effects of perinatal exposure to MXC on reproductive function after maturation in both male and female rats. Pregnant rats were fed a phytoestrogen-free diet containing different doses of MXC (24, 240, and 1200 ppm) from day 15 of gestation to day 10 after parturition, and reproductive functions of offspring were examined after maturation. In males, MXC exposure during the perinatal period decreased serum LH and FSH, but not testosterone levels, but it did not affect copulatory behavior. In females, MXC exposure prolonged the days exhibiting cornified vaginal smears during the estrous cycle. In addition, both the lordosis reflex and preovulatory LH surge on the presumptive proestrous evening were suppressed in MXC-exposed females. These results suggest that perinatal exposure to MXC exerts permanent effects on several aspects of the reproductive function in both male and female rats.     

Time-dependent changes in inhibitory action of lipopolysaccharide on intestinal motility in rat

Endotoxin causes gastrointestinal motility disorder. Aim of this study is to clarify inhibitory mechanisms of lipopolysaccharide (LPS) on smooth muscle contraction in rat ileum. Ileal tissues were isolated from control rat or from LPS-induced peritonitis model rat. Treatment with LPS inhibited carbachol (CCh)-mediated contraction in a time-dependent manner. Cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) genes were also upregulated, but iNOS expression was preceded by a rising of COX-2. All subtypes of prostaglandin E₂ (PGE₂) receptors (EP1-EP4) were expressed in ileum, and PGE₂ and selective EP2 or EP4 agonist inhibited CCh-mediated contraction. Selective iNOS inhibitor did not reverse LPS-induced inhibition of contraction by CCh at 1 and 2 hr, but reduced the inhibitory action at 4 hr after the LPS treatment. COX-2 inhibitor reversed the inhibitory action by LPS in all exposure time. Finally, in ileal tissues isolated from peritonitis model rat, iNOS expression was upregulated only at 4 hr after LPS administration, resulting in enhanced inhibitory action of LPS against CCh-induced contraction. In conclusion, LPS induces COX-2 to produce PGE₂, which initially activates EP2 and/or EP4 on smooth muscle cells to inhibit the contractility in early phase of LPS exposure. Moreover, in late phase of LPS treatment, iNOS is expressed to produce NO, which in turn inhibited the contraction by CCh. The inhibitory cascade is similar in the ileum isolated from peritonitis model rat, indicating time-dependent changes of inhibitory action by LPS on intestinal motility in peritonitis.     

Age-dependent changes in progranulin expression in the mouse brain

The progranulin (PGRN) gene is involved in sexual differentiation of the brain during the perinatal period and estrogen-induced adult neurogenesis in the hippocampus. Mutations in the PGRN gene are also implicated in human frontotemporal lobar degeneration. Thus, while PGRN appears to play important roles as a growth factor in the brain, the localization of PGRN-expressing cells throughout the brain has not been fully established. In the present study, we examined the localization of PGRN proteins in the brain using adult male wild-type mice and PGRN-deficient mice we had generated previously. We also evaluated age-dependent changes in PGRN expression at the mRNA and protein levels. As expected, no immunoreactivity was observed in the brains of the PGRN-deficient mice. In the wild-type mice, intense immunoreactivity was observed in several brain regions including the cingulate and piriform cortices, the pyramidal cell layer and dentate gyrus of the hippocampus, the amygdala, the ventromedial and arcuate nuclei of the hypothalamus and the Purkinje cell layer in the cerebellum. Moreover, PGRN mRNA and protein expression decreased in the cortex, hippocampus and hypothalamus in an age-dependent manner. Since many of these brain regions are involved in emotion, memory and recognition, PGRN may play roles as a growth factor in these brain functions that decline with age.     

Evaluation of krill (Euphausia superba) meal as a partial replacement for fish meal in rainbow trout (Oncorhynchus mykiss) diets

We studied partial replacement of fish meal (FM) by krill meal (KM) and how fluoride from KM enriched-muscles and bones of rainbow trout reared in freshwater affected these fish. Diets that replaced FM with KM at proportions of 0, 7, 15 and 30% (control, KM7, KM15, and KM30) were fed to groups of rainbow trout for 92 days and growth was observed. In fish fed KM7 and KM15, weight gain (WG), feed intake (FI) and specific growth rate (SGR) were unchanged compared with fish fed the control diet, but in fish fed KM30, WG, FI and SGR significantly decreased. After the experiment, fluoride concentration in dorsal muscles of each experimental group was below the detected limit (1 mg/kg), but in vertebral bones, the fluoride concentration increased with each increase in KM inclusion in the diets: 490 mg/kg (control), 755 mg/kg (KM7), 1100 mg/kg (KM15), and 2400 mg/kg (KM30). Tissue specimens of liver of each experimental group had no histopathological changes. Therefore, accumulation of fluoride in vertebral bones apparently adversely affected growth of the vertebral bones thus affecting the growth performance as shown by the decrease in WG, FI and SGR.     

A mesoscale iron enrichment in the western subarctic Pacific induces a large centric diatom bloom

We have performed an in situ test of the iron limitation hypothesis in the subarctic North Pacific Ocean. A single enrichment of dissolved iron caused a large increase in phytoplankton standing stock and decreases in macronutrients and dissolved carbon dioxide. The dominant phytoplankton species shifted after the iron addition from pennate diatoms to a centric diatom, Chaetoceros debilis, that showed a very high growth rate, 2.6 doublings per day. We conclude that the bioavailability of iron regulates the magnitude of the phytoplankton biomass and the key phytoplankton species that determine the biogeochemical sensitivity to iron supply of high-nitrate, low-chlorophyll waters.     

Artificial and natural cross breeding between Atlantic salmon and salmonids currently present in Japan

We evaluated the potential for hybridization between Atlantic salmon and salmonids currently present in Japan. We performed artificial and natural crosses and tested for hybridization using short interspersed elements. We documented the occurrence of hybrids and gynogenesis following artificial crossing. However, the survival rate of these offspring was low (<2 % to the fry stage). We were unable to obtain viable offspring from natural crosses of Atlantic salmon and chum salmon or masu salmon. Our results suggest that there is little risk of hybridization between Atlantic salmon and salmonids that are currently present in the wild in Japan. Furthermore, our results suggest that gynogenesis may occur naturally.     

Effect of EMC and air in wood on the new in-process moisture content monitoring concept under radiofrequency/vacuum (RF/V) drying

To improve the accuracy of the new in-process moisture content (MC) monitoring concept under radiofrequency/vacuum (RF/V) drying, equilibrium moisture content (EMC) tests were carried out under various ambient pressures, and pressure curves in wood were analyzed during chamber evacuation and heating phases. The results showed that EMC increased with a decrease in ambient pressure regardless of temperature, relative humidity (RH), and species. The accuracy of MC estimation for Hinoki under RF/V drying was improved from 1.5% maximum absolute errors to 0.6% after EMC modification. The pressure curves for Hinoki and Sugi under RF/V drying showed similar tendencies to an idealized process. Russian larch showed different curves, indicating that the pressure in the wood did not reach the ambient pressure because of its low permeability. Therefore, MC could not be estimated using this monitoring concept because of the presence of much air in the wood of Russian larch.