Effect of seed coat color on seed dormancy in different environments

Arachis glabrata Benth, variety glabrata coll. GK 10596 (PI 276233; ICG8176) belonging to section Rhizomatosae has multiple disease resistance. Fertile hybrids between A. hypogaea L. and A. glabrata, A. duranensis Krapov & W.C. Gregory and A. glabrata and A. diogoi Hoehne and A. glabrata were produced. Introgression of DNA from A. glabrata into A. hypogaea, A. duranensis and A. diogoi was analyzed by isozyme and RAPD analyses. Hybrids were backcrossed and BC₁ seeds were obtained in all the three hybrids. Hybrids were evaluated for the transfer of disease resistance genes from A. glabrata, which was confirmed. RAPD analysis with several primers showed that DNA fragment pattern were not simply represented, instead there were new bands and several parental bands were absent in the interspecific derivatives. This revised version was published online in August 2006 with corrections to the Cover Date.     

Involvement of Gonadotropin-Releasing Hormone in Thyroxine Release in Three different forms of Teleost Fish: Barfin Founder, Masu Salmon and Goldfish

Effects of gonadotropin-releasing hormone (GnRH) on thyroxine (T₄) release in vivo and in vitro were studied in barfin flounder Verasper moseri, masu salmon Oncorhynchus masou and goldfish Carassius auratus. Seabream GnRH (sbGnRH) at a dose of 200 ng/50 g body weight (BW) significantly increased plasma T₄ levels 1 h after the in vivo injection in the barfin flounder, but thereafter the levels normalized. Salmon GnRH (sGnRH) significantly increased plasma T₄ levels l h after the injection with a significant return to initial levels in male masu salmon and male goldfish. In contrast, sGnRH and cGnRH-II in barfin flounder, and cGnRH-II in male masu salmon and male goldfish were not effective in stimulating T₄ release. To clarify direct involvement of GnRH in T₄ release, dissected lower jaw including scattered thyroid follicles was incubated with sbGnRH (1 μg/well) in barfin flounder, and with two doses (0.1 and 1 μg/well) of sGnRH in masu salmon and goldfish in vitro. T₄ concentrations of control were stable during 24 h. Incubation of lower jaw with high dose (1 μg/well) of GnRH significantly (P<0.05) increased T₄ concentrations of incubation medium at 1 h in all experimental fishes. These results indicate that direct stimulation of T₄ secretion by GnRH occurs widely in teleost fish.     

Photic and circadian regulation of self-feeding activity in ayu

ABSTRACT:   The self-feeding rhythms of ayu Plecoglossus altivelis altivelis were examined. Individual ayu (mean body weight 40 g) were held in 60-L glass tanks equipped with self-feeders. Six of 14 fish learned self-feeding during the experiment. Under two different light–dark (LD) conditions (16 h:8 h and 8 h:16 h LD), self-feeding was synchronized to the LD cycle, and feeding occurred almost exclusively during the light phase. During exposure to constant light (LL), circadian feeding rhythms were observed. These results indicate self-feeding rhythms in ayu are restricted to the light phase under LD conditions and are controlled by the circadian clock under LL conditions.     

Toxicity of wild juvenile “komonfugu” Takifugu flavipterus in the Bay of Sanriku Coast,Tohoku Area,Northern Japan

To clarify the mechanism of tetrodotoxin (TTX) accumulation in pufferfish, we compared the toxicity of two sets of wild juvenile “komonfugu” Takifugu flavipterus. The first set was sampled from Onisawa Fishing Port (FP) located in Okirai Bay, the Pacific Coast of Sanriku, Tohoku Area, Northern Japan. The second set was collected from the Onisawa FP and reared in an outdoor laboratory tank supplied with different seawater (Yoshihama Bay). The fish were sampled regularly and on the same days. The amount of TTX (mouse unit (MU)/fish) in the fish at Onisawa FP increased until 20 days and thereafter it did not change, while the amount of TTX in the fish in the laboratory tank remained low, and the TTX concentration (MU/g fish) decreased. Next, we compared the toxicity of wild juvenile T. flavipterus collected from Okirai Bay (Onisawa FP and Okirai FP) and Yoshihama Bay (Yoshihama FP). Large differences in TTX levels were observed among the fish from the three FPs. The amounts and concentrations of TTX in the fish at Onisawa FP were higher than those in the fish from the other two FPs. These results indicate that a large variation in toxic activity exists in the juvenile T. flavipterus in the bay of the Sanriku Coast.

     

Inhibition activity of essential oils obtained from Japanese trees against <Emphasis Type="Italic">Skeletonema costatum</Emphasis>

The growth inhibition activities of essential oils obtained from Cryptomeria japonica, Chamaecyparis obtusa, and Pinus thunbergii were examined against the bacillariophyceae Skeletonema costatum, also known as red tide plankton. The essential oils were extracted from the heartwood, leaves, and bark of these typical indigenous Japanese conifers. The essential oils from C. japonica bark and P. thunbergii heartwood possessed strong growth inhibition activity. The chemical compositions of these essential oils were analyzed by gas chromatography/flame ionization detection (GC-FID) and gas chromatography/mass spectrometry (GC-MS). α-Terpineol and longifolene were the main components of the essential oil from P. thunbergii heartwood. The C. japonica bark essential oil was mainly composed of α-terpineol, δ-cadinene, isophyllocladene, and ferruginol. Ferruginol and longifolene showed more potent growth inhibition against S. costatum than hinokitiol (β-thujaplicine), which is known to be a strong antifungal compound among wood components. Ferruginol and longifolene were important factors for the growth inhibition activity of the essential oils from C. japonica bark and P. thunbergii heartwood, respectively. These results suggest the possibility of using C. japonica bark and P. thunbergii heartwood for the control of red tide plankton.     

Increase of cells expressing PD-L1 in bovine leukemia virus infection and enhancement of anti-viral immune responses in vitro via PD-L1 blockade

ABSTRACT: The inhibitory receptor programmed death-1 (PD-1) and its ligand, programmed death-ligand 1 (PD-L1) are involved in immune evasion mechanisms for several pathogens causing chronic infections. Blockade of the PD-1/PD-L1 pathway restores anti-virus immune responses, with concomitant reduction in viral load. In a previous report, we showed that, in bovine leukemia virus (BLV) infection, the expression of bovine PD-1 is closely associated with disease progression. However, the functions of bovine PD-L1 are still unknown. To investigate the role of PD-L1 in BLV infection, we identified the bovine PD-L1 gene, and examined PD-L1 expression in BLV-infected cattle in comparison with uninfected cattle. The deduced amino acid sequence of bovine PD-L1 shows high homology to the human and mouse PD-L1. The proportion of PD-L1 positive cells, especially among B cells, was upregulated in cattle with the late stage of the disease compared to cattle at the aleukemic infection stage or uninfected cattle. The proportion of PD-L1 positive cells correlated positively with prediction markers for the progression of the disease such as leukocyte number, virus load and virus titer whilst on the contrary, it inversely correlated with the degree of interferon-gamma expression. Blockade of the PD-1/PD-L1 pathway in vitro by PD-L1-specific antibody upregulated the production of interleukin-2 and interferon-gamma, and correspondingly, downregulated the BLV provirus load and the proportion of BLV-gp51 expressing cells. These data suggest that PD-L1 induces immunoinhibition in disease progressed cattle during chronic BLV infection. Therefore, PD-L1 would be a potential target for developing immunotherapies against BLV infection.     

Molecular phylogeny and diversity of Myanmar and Bhutan mithun based on mtDNA sequences

The mithun (Bos frontalis), synonymous with mithan and gayal, is considered to be a domesticated form of gaur (B. gaurus). However, there has been a controversy concerning its origin. In an effort to address this issue, the mitochondrial cytochrome b (cytb) genes of 20 mithun from Myanmar and 13 from Bhutan were sequenced to trace its maternal origin. Seven cytb haplotypes were found in the 33 mithun, and the phylogenetic tree for these haplotypes clearly showed three embranchments involving five gaur types, a B. indicus type, and a B. taurus type. Sixteen Myanmar and 12 Bhutan mithun had gaur haplotypes, while a B. indicus haplotype was found in three Myanmar and one Bhutan mithun. The B. taurus haplotype was detected in a single Myanmar animal. These results demonstrated that the principal maternal origin of mithun was gaur and suggested that it was directly domesticated from gaur. However, some introgression of domestic cattle existed in current mithun populations. The presence of cattle mtDNA raised the question of how many cattle nuclear genes might have been integrated into the gene pool of mithun.     

Effects of reactive radicals and heat on <Emphasis Type="Italic">trans</Emphasis>-isomerization of eicosapentaenoic acid

Trans geometric isomers of eicosapentaenoic acid (TEPA) have been found as minor components in human platelets. However, there is little information on the mechanism of trans-isomerization of eicosapentaenoic acid (EPA) in vitro and in vivo. The effects of reactive radicals and heat on trans-isomerization of EPA were examined. Trans-isomerization occurred when EPA ethyl ester reacted with nitrogen dioxide radical (NO₂) but not with 2,2′-azobis(2,4-dimethylvaleronitrile). TEPA was also produced from EPA ethyl ester heated at 200°C for 60 h. No TEPA, however, was detected in sardines Sardinops melanostictus after boiling, roasting, or microwave heating. These results suggest that EPA is trans-isomerized by NO₂ in vivo while trans-isomerization of EPA does not occur during conventional cooking.     

Increased bovine Tim-3 and its ligand expressions during bovine leukemia virus infection

ABSTRACT: The immunoinhibitory receptor T cell immunoglobulin domain and mucin domain-3 (Tim-3) and its ligand, galectin-9 (Gal-9), are involved in the immune evasion mechanisms for several pathogens causing chronic infections. However, there is no report concerning the role of Tim-3 in diseases of domestic animals. In this study, cDNA encoding for bovine Tim-3 and Gal-9 were cloned and sequenced, and their expression and role in immune reactivation were analyzed in bovine leukemia virus (BLV)-infected cattle. Predicted amino acid sequences of Tim-3 and Gal-9 shared high homologies with human and mouse homologues. Functional domains, including tyrosine kinase phosphorylation motif in the intracellular domain of Tim-3 were highly conserved among cattle and other species. Quantitative real-time PCR analysis showed that bovine Tim-3 mRNA is mainly expressed in T cells such as CD4+ and CD8+ cells, while Gal-9 mRNA is mainly expressed in monocyte and T cells. Tim-3 mRNA expression in CD4+ and CD8+ cells was upregulated during disease progression of BLV infection. Interestingly, expression levels for Tim-3 and Gal-9 correlated positively with viral load in infected cattle. Furthermore, Tim-3 expression level closely correlated with up-regulation of IL-10 in infected cattle. The expression of IFN-γ and IL-2 mRNA was upregulated when PBMC from BLV-infected cattle were cultured with Cos-7 cells expressing Tim-3 to inhibit the Tim-3/Gal-9 pathway. Moreover, combined blockade of the Tim-3/Gal-9 and PD-1/PD-L1 pathways significantly promoted IFN-γ mRNA expression compared with blockade of the PD-1/PD-L1 pathway alone. These results suggest that Tim-3 is involved in the suppression of T cell function during BLV infection.