Nonomuraea maheshkhaliensis sp. nov., a novel actinomycete isolated from mangrove rhizosphere mud

A strain of Nonomuraea was isolated from Maheshkhali, Cox's Bazar, an unexplored region of Bangladesh. Strain 16-5-14(T) is a Gram-positive, aerobic, non-motile actinomycete that formed branched substrate and aerial mycelia. On the basis of 16S rRNA gene sequence similarity studies, strain 16-5-14(T) was shown to belong to the genus Nonomuraea, being most closely related to Nonomuraea kuesteri. Chemotaxonomic data supported allocation of the strain as a member of the genus Nonomuraea. The strain 16-5-14(T) contained MK-9(H(4)) as the major menaquinone, the polar lipid was phosphatidylethanolamine and major cellular fatty acids were observed as C(16 : 0 )(15.5%), iso-C(16 : 0) (13.8%) and 10-methyl C(17 : 0) (9.6%). Results of DNA-DNA hybridization and physiological tests allowed genotypic and phenotypic differentiation of strain 16-5-14(T) from closely related species N. kuesteri. Thus 16-5-14(T) represents a novel species of the genus Nonomuraea. On the basis of evaluation of the morphological, physiological and chemotaxonomic characteristics, 16S rRNA gene sequence comparisons and DNA-DNA hybridization, Nonomuraea maheshkhaliensis sp. nov. (type strain, 16-5-14(T)=JCM 13929(T)=MTCC 8545(T)) is proposed.     

CD83 expression is a sensitive marker of activation required for B cell and CD4+ T cell longevity in vivo

CD83 is a surface marker that differentiates immature and mature human dendritic cell populations. Thymic epithelial cell expression of CD83 is also necessary for efficient CD4+ T cell development in mice. The altered phenotypes of peripheral B and CD4+ T cells, and the reduction of peripheral CD4+ T cells in CD83-/- mice, suggest additional functions for CD83. To assess this, a panel of mAbs was generated to characterize mouse CD83 expression by peripheral leukocytes. As in humans, activation of conventional and plasmacytoid murine dendritic cell subsets led to rapid up-regulation of CD83 surface expression in mice. In primary and secondary lymphoid compartments, a subset of B cells expressed low-level CD83, while CD83 was not detected on resting T cells. However, CD83 was prominently up-regulated on the majority of spleen B and T cells within hours of activation in vitro. In vivo, a low dose of hen egg lysozyme (1 microg) induced significant CD83 but not CD69 expression by Ag-specific B cells within 4 h of Ag challenge. Although B cell development appeared normal in CD83-/- mice, B and CD4+ T cell expression of CD83 was required for lymphocyte longevity in adoptive transfer experiments. Thus, the restricted expression pattern of CD83, its rapid induction following B cell and T cell activation, and its requirement for B cell and CD4+ T cell longevity demonstrate that CD83 is a functionally significant and sensitive marker of early lymphocyte activation in vivo.     

Rapid down-regulation of Ret following exposure of dopaminergic neurons to neurotoxins

The survival and functional maintenance of vertebrate neurons depends on the availability of specific neurotrophic factors. We studied the influence of neurotrophic support on responses of dopaminergic neurons to 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine, a neurotoxin known to damage the nigrostriatal dopaminergic pathway in humans and other mammals. Treatment of mice with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine caused decreases in levels of Ret, a tyrosine kinase receptor for glial cell line-derived neurotrophic factor (GDNF) in the striatum, under the condition in which tyrosine hydroxylase was moderately decreased and the GDNF family receptor alpha1, another receptor of GDNF that is the ligand-binding subunit, were unaffected. Down-regulation of Ret was also observed in dopamine-producing PC12 cells undergoing apoptosis induced by rotenone, another toxic substance for dopaminergic neurons, while other cellular components were not affected. Ret was also extremely vulnerable to other apoptotic inducing conditions. Taken together, these results indicate that Ret, an important signal molecule in dopaminergic neurons, may be down-regulated in the early stages of neuronal degeneration caused by various neurotoxic substances, and may lead to reduced neurotrophic influences.     

A mutation on influenza C virus M1 protein affects virion morphology by altering the membrane affinity of the protein

Reverse genetics has been documented for influenza A, B, and Thogoto viruses belonging to the family Orthomyxoviridae. We report here the reverse genetics of influenza C virus, another member of this family. The seven viral RNA (vRNA) segments of C/Ann Arbor/1/50 were expressed in 293T cells from cloned cDNAs, together with nine influenza C virus proteins. At 48 h posttransfection, the infectious titer of the culture supernatant was determined to be 2.51 x 10(3) 50% egg infectious doses/ml, which is lower than the number of influenza C virus-like particles (VLPs) (10(6)/ml) generated using the same system. By generating influenza C VLPs containing a given vRNA segment, we showed that each of the vRNA segments was similarly synthesized in the plasmid-transfected cells but that some segments were less efficiently incorporated into the VLPs. This finding leads us to speculate that the differences in incorporation efficiency into VLPs between segments might be a reason for the inefficient production of infectious viruses. Second, we generated a mutant recombinant virus, rMG96A, which possesses an Ala-->Thr mutation at residue 24 of the M1 protein, a substitution demonstrated to be involved in the morphology (filamentous or spherical) of the influenza C VLPs. As expected, rMG96A exhibited a spherical morphology, whereas recombinant wild-type of C/Ann Arbor/1/50, rWT, exhibited a mainly filamentous morphology. Membrane flotation analysis of the cells infected with rWT or rMG96A revealed a difference in the ratio of membrane-associated M1 proteins, suggesting that the affinity of M1 protein to the cell membrane is a determinant for virion morphology.     

Estrogen prevents bone loss via estrogen receptor alpha and induction of Fas ligand in osteoclasts

Estrogen prevents osteoporotic bone loss by attenuating bone resorption; however, the molecular basis for this is unknown. Here, we report a critical role for the osteoclastic estrogen receptor alpha (ERalpha) in mediating estrogen-dependent bone maintenance in female mice. We selectively ablated ERalpha in differentiated osteoclasts (ERalpha(DeltaOc/DeltaOc)) and found that ERalpha(DeltaOc/DeltaOc) females, but not males, exhibited trabecular bone loss, similar to the osteoporotic bone phenotype in postmenopausal women. Further, we show that estrogen induced apoptosis and upregulation of Fas ligand (FasL) expression in osteoclasts of the trabecular bones of WT but not ERalpha(DeltaOc/DeltaOc) mice. The expression of ERalpha was also required for the induction of apoptosis by tamoxifen and estrogen in cultured osteoclasts. Our results support a model in which estrogen regulates the life span of mature osteoclasts via the induction of the Fas/FasL system, thereby providing an explanation for the osteoprotective function of estrogen as well as SERMs.     

Transforming growth factor-beta in the brain is activated by exercise and increases mobilization of fat-related energy substrates in rats

We have recently reported that inhibition of transforming growth factor (TGF)-beta in the brain reduced fat-related energy substrates concentrations in response to exercise. We investigated the relevance between the mobilization of fat-related energy substrates (nonesterified fatty acid and ketone bodies) during exercise and the effects of TGF-beta in the brain. Low-intensity exercise was simulated by contraction of the hindlimbs, induced by electrical stimulation at 2 Hz in anesthetized rats (Sim-Ex). As with actual exercise, it was confirmed that mobilization of carbohydrate-related energy substrates (glucose and lactic acid) occurred immediately after the onset of Sim-Ex, and mobilization of fat-related energy substrates followed thereafter. The timing of mobilization of fat-related substrates corresponded to that of the increase in TGF-beta in cerebrospinal fluid (CSF) in Sim-Ex. The level of TGF-beta in CSF significantly increased after 10 min of Sim-Ex and remained elevated until 30 min of Sim-Ex. Intracisternal administration of TGF-beta caused rapid mobilization of fat-related energy substrates. Meanwhile, there were no effects on the changes in carbohydrate-related substrates. The levels of catecholamines were slightly elevated after TGF-beta administration, and, although not significantly in statistical terms, we consider that at least a part of TGF-beta signal was transducted via the sympathetic nervous system because of these increases. These data indicate that TGF-beta in the brain is closely related to the mobilization of fat-related energy substrates during low-intensity exercise. We hypothesized that the central nervous system plays a role in the regulation of energy metabolism during low-intensity exercise and this may be mediated by TGF-beta.