Ancestry of American polyploid Hordeum species with the I genome inferred from 5S and 18S-25S rDNA

* BACKGROUND AND AIMS: The genus Hordeum exists at three ploidy levels (2x, 4x and 6x) and presents excellent material for investigating the patterns of polyploid evolution in plants. Here the aim was to clarify the ancestry of American polyploid species with the I genome. * METHODS: Chromosomal locations of 5S and 18S-25S ribosomal RNA genes were determined by fluorescence in situ hybridization (FISH). In both polyploid and diploid species, variation in 18S-25S rDNA repeated sequences was analysed by the RFLP technique. * KEY RESULTS: Six American tetraploid species were divided into two types that differed in the number of rDNA sites and RFLP profiles. Four hexaploid species were similar in number and location of both types of rDNA sites, but the RFLP profiles of 18S-25S rDNA revealed one species, H. arizonicum, with a different ancestry. * CONCLUSIONS: Five American perennial tetraploid species appear to be alloploids having the genomes of an Asian diploid H. roshevitzii and an American diploid species. The North American annual tetraploid H. depressum is probably a segmental alloploid combining the two closely related genomes of American diploid species. A hexaploid species, H. arizonicum, involves a diploid species, H. pusillum, in its ancestry; both species share the annual growth habit and are distributed in North America. Polymorphisms of rDNA sites detected by FISH and RFLP analyses provide useful information to infer the phylogenetic relationships of I-genome Hordeum species because of their highly conserved nature during polyploid evolution.     

Species-specific TNF induction of thymocyte proliferation

Summary Recombinant murine (rMu) tumor necrosis factor (TNF), in a standard comitogenic assay with phytohemagglutinin, induced murine thymocyte proliferation, while up to 10,000-fold higher concentrations of recombinant human TNF did not. The induction of thymocyte proliferation was dependent upon TNF concentration in a biphasic manner. Thus, 100 to 1000 units/ml TNF were near optimal while concentrations 1,000 units/ml caused apparent down regulation. The effect was abrogated by neutralizing antibody to rMu-TNF but not by neutralizing antibody to rMu-interleukin 1 or . The rMu-TNF did not induce proliferation of the mature murine T-helper cell line, D10.G4.1, in the presence of mitogen. Taken together the results indicate that TNF, in a strictly species-specific manner, can regulate thymocyte proliferation independently of interleukin 1.Supported in part by Asahi Chemical Industry Co., Inc. and by USPHS Grants CA-24538, CA-15142 and CA-09072 awarded by the National Cancer Institute, Department of Health and Human Services     

Chronic exposure to an extremely low‐frequency magnetic field induces depression‐like behavior and corticosterone secretion without enhancement of the hypothalamic–pituitary–adrenal axis in mice

An extremely low‐frequency magnetic field (ELF‐MF) is generated by power lines and household electrical devices. Many studies have suggested an association between chronic ELF‐MF exposure and anxiety and/or depression. The mechanism of these effects is assumed to be a stress response induced by ELF‐MF exposure. However, this mechanism remains controversial. In the present study, we investigated whether chronic ELF‐MF exposure (intensity, 3 mT; total exposure, 200 h) affected emotional behavior and corticosterone synthesis in mice. ELF‐MF‐treated mice showed a significant increase in total immobility time in a forced swim test and showed latency to enter the light box in a light–dark transition test, compared with sham‐treated (control) mice. Corticosterone secretion was significantly high in the ELF‐MF‐exposed mice; however, no changes were observed in the amount of the adrenocorticotropic hormone and the expression of genes related to stress response. Quantification of the mRNA levels of adrenal corticosteroid synthesis enzymes revealed a significant reduction in Cyp17a1 mRNA in the ELF‐MF‐exposed mice. Our findings suggest the possibility that high intensity and chronic exposure to ELF‐MF induces an increase in corticosterone secretion, along with depression‐ and/or anxiety‐like behavior, without enhancement of the hypothalamic–pituitary–adrenal axis. Bioelectromagnetics 34:43–51, 2013. © 2012 Wiley Periodicals, Inc.     

Alterations in hepatitis B virus nucleotide sequences in a chronic virus carrier from immunotolerant to immunoactive phase

Factors involved in transition from the immunotolerant to immunoactive phase in chronic hepatitis B virus (HBV) infection remain unclear. We investigated viral mutations occurring during transition and elucidated their virological and immunological significance. Full-length HBV DNA sequences were serially determined in a chronic HBV carrier from the immunotolerant to immunoactive phase. Viral replicative competence was examined by transfection analysis. HBV-specific CD8⁺ T cell response was evaluated by coculture of CD8⁺ T cells with autologous dendritic cells followed by interferon-γ Elispot assay. Eleven point mutations and two deletions appeared around the onset of the immunoactive phase. Viral replicative competence declined significantly after the onset of active hepatitis. Examination of the CD8⁺ T cell response against two putative T-cell epitopes, which contained substituted amino acids from the immunotolerant to immunoactive phase, showed that mutant HBV epitopes gave a lesser T cell response than wild-type HBV ones. In summary, point mutations and deletions may occur prior to or concurrent with the onset of the immunoactive phase during chronic HBV infection. These mutations may result in a significant decrease in both viral replicative competence and HBV-specific CD8⁺ T cell response, suggesting a possible adaptation for the maintenance of viral persistence.     

MHC class I A loci polymorphism and diversity in three Southeast Asian populations of cynomolgus macaque

Cynomolgus macaques (Macaca fascicularis, Mafa) have emerged as important animal models for biomedical research, necessitating a more extensive characterization of their major histocompatibility complex polymorphic regions. The current information on the polymorphism or diversity of the polygenetic Mafa class I A loci is limited in comparison to the more commonly studied rhesus macaque Mafa class I A loci. Therefore, in this paper, to better elucidate the degree and types of polymorphisms and genetic differences of Mafa-A1 among three native Southeast Asian populations (Indonesian, Vietnamese, and Filipino) and to investigate how the allele differences between macaques and humans might have evolved to affect their respective immune responses, we identified 83 Mafa-A loci-derived alleles by DNA sequencing of which 66 are newly described. Most alleles are unique to each population, but seven of the most frequent alleles were identical in sequence to some alleles in other macaque species. We also revealed (1) the large and dynamic genetic and structural differences and similarities in allelic variation by analyzing the population allele frequencies, Hardy-Weinberg’s equilibrium, heterozygosity, nucleotide diversity profiles, and phylogeny, (2) the difference in genetic structure of populations by Wright’s FST statistic and hierarchical analysis of molecular variance, and (3) the different demographic and selection pressures on the three populations by performing Tajima’s D test of neutrality. The large level of diversity and polymorphism at the Mafa-A1 was less evident in the Filipino than in the Vietnam or the Indonesian populations, which may have important implications in animal capture, selection, and breeding for medical research.     

Isolation and Structure of Neoannonin,a Novel Insecticidal Compound from the Seeds of Annona squamosa

Two insecticidal compounds were isolated from the seeds of Annona squamosa. One was identified as annonin (squamocin) and the other was characterized as a novel dihydroxy-bistetrahydrofuran fatty acid lactone (acetogenin) with 35 carbons. These two compounds were toxic to eggs, larvae and adults of the fruitfly.     

Relationships between stretch-shortening cycle performance and maximum muscle strength

This study aimed to examine the relationships between muscle power output using the stretch-shortening cycle (SSC) and maximum strength, as measured by the 1 RM (1 repetition maximum) test and the isokinetic dynamometer under elbow flexion. Sixteen trained, young adult males pulled a constant load of 40% MVC (maximum voluntary elbow flexion contraction) by ballistic elbow flexion under the following two preliminary conditions: 1) the static relaxed muscle state (SR condition) and 2) using the SSC (SSC condition). Muscle power was determined from the product of the pulling velocity and load mass by a power measurement instrument with a rotary encoder. The 1 RM bench press (1RM BP) and isokinetic maximum strength under elbow flexion with the Cybex-325 were measured as indicators of dynamic maximum strength. 1) The early power output exerted under the SSC condition showed a significant and high correlation with the 1 RM BP (r = 0.83), but only moderate correlation with the isokinetic muscle strength (r = 0.50-0.67). 2) The contribution of the 1 RM BP to the early muscle contraction velocity exerted under the SSC condition was large. These results suggested that muscle power exerted using the SSC shows a stronger relationship with maximum muscle strength measured by a 1 RM test rather than isokinetic maximum strength.     

The orexigenic effect of GnIH is mediated by central opioid receptors in chicks

Gonadotropin-inhibiting hormone (GnIH) is a newly discovered hypothalamic hormone which suppresses gonadotropin synthesis and release from the anterior pituitary. Recently, we found that intracerebroventricular (ICV) injection of GnIH stimulated feeding behavior of chicks (Gallus gallus) and suggested that GnIH is one of orexigenic peptides. However, the mechanism underlying the orexigenic effect is still unknown. In the present study, we examined whether the orexigenic effect of GnIH is related to opioid and nitric oxide (NO) systems. The orexigenic effect of ICV-injected GnIH was attenuated by co-injection of beta-funaltrexamine (an opioid mu-receptor antagonist) but not ICI-174,864 and nor-binaltorphimine (antagonists of opioid delta- and kappa-receptors, respectively). The co-injection of non-selective NO synthase inhibitor did not affect GnIH-induced feeding behavior. The present study demonstrated that the GnIH-induced feeding might be mediated by opioid mu-receptor in chicks.     

Presence of xyloglucan-like polysaccharide in Spirogyra and possible involvement in cell–cell attachment

By methylation analysis, it was found that the cell walls of Spirogyra contained 4,6-linked glucose, 4-linked glucose and terminal xylose, which could be components of xyloglucan. Immunocytochemical analysis was carried out using an anti-serum against xyloglucan. After removal of pectic substances, the cell walls of both rhizoid cells and inner cells were stained. Crude protein extract from Spirogyra had a hydrolase activity for xyloglucans. In addition, the exogenously applied xyloglucan prevented the detachment of the cell wall of the severed cell. Involvement of xyloglucan-like polysaccharide in cell–cell attachment was discussed.     

Conformation-activity relationship on novel 4-pyridylmethylthio derivatives with antiangiogenic activity

We found 4-pyridylmethylthio derivative 1 to be very effective in using antiangiogenesis activity to prevent proliferation of HUVECs (Human Umbilical Vein Endothelial Cells), which was induced by vascular endothelial growth factor (VEGF). Compound 1 was equally effective in inhibiting VEGF receptor2 tyrosine kinase (KDR, IC₅₀ = 26 nM). We deduced that the inhibition was the result of binding the catalytic domain of VEGF receptor2 tyrosine kinase in a similar fashion to both phthalazine derivative PTK787 2 and anthranylamide derivative AAL993 3. In this report, we will describe the conformational analyses, from ab initio MO calculation and X-ray crystallographic analyses, of compound 1 and the analogs, which include non-active 9, all in comparison with 2 and 3. The conformation–activity relationships suggest that a nonbonded intramolecular interaction between the sulfur and the carbonyl oxygen of 1 was very important in inhibiting KDR.