Genome sequence of extracellular-protease-producing Alishewanella jeotgali isolated from traditional Korean fermented seafood

Alishewanella jeotgali MS1(T) (= KCTC 22429(T) = JCM 15561(T)) was isolated from a traditional Korean fermented seafood, gajami sikhae (jeotgal), and has been reported as a novel species. A. jeotgali was proven to have extracellular proteolytic activity, which may play an important role in the fermentation environment of food containing fish flesh. Here, we present the genome sequence of Alishewanella jeotgali MS1(T) as the first sequenced strain in the genus Alishewanella and its taxonomic relatives.     

Genome Sequence of Pectin-Degrading Alishewanella aestuarii Strain B11T, Isolated from Tidal Flat Sediment

We present the genome sequence of Alishewanella aestuarii B11(T) (=KCTC 22051(T)=DSM 19476(T)). This species, isolated from tidal flat sediment, was reported to be a novel species. A. aestuarii is known to degrade pectin, an important component of plant cell wall. The presence of the genes related to pectin metabolism in this strain indicates its capability to utilize pectin.     

T-helper cell and associated antibody response to synthetic peptides of the E glycoprotein of Murray Valley encephalitis virus.

A battery of 16 synthetic peptides, selected primarily by computer analysis for predicted B- and T-cell epitopes, was prepared from the deduced amino acid sequence of the envelope (E) glycoprotein of Murray Valley encephalitis (MVE) virus. We examined all of the peptides for T-helper (Th)-cell recognition and antibody induction in three strains of mice: C57BL/6, BALB/c, and C3H. Lymphoproliferative and interleukin-2 assays were performed on splenic T cells from mice inoculated with peptides in Freund's incomplete adjuvant or with MVE virus. Several peptides found to contain predicted T-cell epitopes elicited a Th-cell response in at least one strain of mice, usually with a concomitant antibody response. Peptides 145 (amino acids 145 to 169) and 17 (amino acids 356 to 376) were strongly recognized by T cells from all three inbred strains of mice. Peptide 06 (amino acids 230 to 251) primed C57BL/6 mice for Th- and B-cell reactivity with native MVE virus, and T cells from virus-immune mice were stimulated by this peptide. Peptide 06 was recognized by several Th-cell clones prepared from mice immunized with MVE, West Nile, or Kunjin virus. These results indicate that it may be feasible to design synthetic flavivirus peptides that define T-cell epitopes capable of generating a helper cell response for B-cell epitopes involved in protective immunity.     

Draft genome sequence of Pedobacter agri PB92T, which belongs to the family Sphingobacteriaceae

Strain PB92(T) of Pedobacter agri, which belongs to the family Sphingobacteriaceae, was isolated from soil in the Republic of Korea. The draft genome of strain PB92(T) contains 5,141,552 bp, with a G+C content of 38.0%. This is the third genome sequencing project of the type strains among the Pedobacter species.     

Biosynthesis of brassinolide from teasterone via typhasterol and castasterone in cultured cells of Catharanthus roseus

The biosynthesis of brassinolide (BL) in crown gall and nontransformed cells of Catharanthus roseus in which BL, castasterone (CS), typhasterol (TY), and teasterone (TE) are endogenous was investigated using deuterated TY and TE as substrates. The metabolites were analyzed by gas chromatography-mass spectrometry (GC-MS) and/or GC-selected ion monitoring (SIM). It was found that these cells converted TY to CS and BL, as well as TE to TY and CS. Because the pathway from CS to BL in the cells has already been confirmed, a biosynthetic sequence of TE TY CS BL was established. Reversible conversion between TE and TY was observed.Biosynthesis of brassinosteroids in Catharanthus roseus. Part III. Part II of this series: Suzuki et al. (1993).     

Lithium: Short-term and chronic effects on plasma testosterone and luteinizing hormone concentrations in mice

The effects of short-term and chronic lithium administration on the concentrations of plasma testosterone (T) and luteinizing hormone (LH) were evaluated in C57BL/6 mice, maintained on a fixed photo-period of LD 14:10 (white lights on at 06:00 h, CST). Lithium chloride was injected intraperitoneally twice daily (at 09:00 and 16:00 h) in groups of adult male mice at a dosage of 2.5 meq/kg for 7 days, and 1.25 meq/kg for 21 days. Circulating levels of T and LH were measured by standard radioimmunoassay (RIA) methods. Plasma T levels showed a significant increase in mice treated with lithium for 7 days as compared to those in saline-injected control animals. However, there was no significant difference in the concentrations of plasma T between chronic (21 days) lithium-treated mice and the matched control. Plasma LH levels remained unchanged following both short-term and chronic lithium treatment.     

Absolute Sustainability‐Based Life Cycle Assessment (ASLCA): A Benchmarking Approach to Operate Agri‐food Systems within the 2°C Global Carbon Budget

Given the increasing environmental impacts associated with global agri‐food systems, operating and developing these systems within the so‐called absolute environmental boundaries has become crucial, and hence the absolute environmental sustainability concept is particularly relevant. This study introduces an approach called absolute sustainability‐based life cycle assessment (ASLCA) that informs the climate impacts of an agri‐food system (on any economic level) in absolute terms. First, a global carbon budget was calculated that is sufficient to limit global warming to below 2°C. Next, a share of the carbon budget available to the global agri‐food sector was estimated, and then it was shared between agri‐food systems on multiple economic levels using four alternative methods. Third, the climate impacts of those systems were calculated using life cycle assessment methodology and were benchmarked against those carbon budget shares. This approach was used to assess a number of New Zealand agri‐food systems (agri‐food sector, horticulture industries and products) to investigate how these systems operated relative to their carbon budget shares. The results showed that, in 2013, the New Zealand agri‐food systems were within their carbon budget shares for one of the four methods, and illustrated the scale of change required for agri‐food systems to perform within their carbon budget shares. This method can potentially be extended to consider other environmental impacts with global boundaries; however, further development of the ASLCA is necessary to account for other environmental impacts whose boundaries are only meaningful when defined at a regional or local level.     

Coordinated function of murine cytomegalovirus genes completely inhibits CTL lysis

Murine CMV (MCMV) encodes three viral genes that interfere with Ag presentation (VIPRs) to CD8 T cells, m04, m06, and m152. Because the functional impact of these genes during normal infection of C57BL/6 mice is surprisingly modest, we wanted to determine whether the VIPRs are equally effective against the entire spectrum of H-2(b)-restricted CD8 T cell epitopes. We also wanted to understand how the VIPRs interact at a functional level. To address these questions, we used a panel of MCMV mutants lacking each VIPR in all possible combinations, and CTL specific for 15 H-2(b)-restricted MCMV epitopes. Only expression of all three MCMV VIPRs completely inhibited killing by CTL specific for all 15 epitopes, but removal of any one VIPR enabled lysis by at least some CTL. The dominant interaction between the VIPRs was cooperation: m06 increased the inhibition of lysis achieved by either m152 or m04. However, for 1 of 15 epitopes m04 functionally antagonized m152. There was little differential impact of any of the VIPRs on K(b) vs D(b), but a surprising degree of differential impact of the three VIPRs for different epitopes. These epitope-specific differences did not correlate with functional avidity, or with timing of VIPR expression in relation to Ag expression in the virus replication cycle. Although questions remain about the molecular mechanism and in vivo role of these genes, we conclude that the coordinated function of MCMV's three VIPRs results in a powerful inhibition of lysis of infected cells by CD8 T cells.     

Proteomics as a complementary tool for identifying unintended side effects occurring in transgenic maize seeds as a result of genetic modifications

To improve the probability of detecting unintended side effects during maize gene manipulations by bombardment, proteomics was used as an analytical tool complementary to the existing safety assessment techniques. Since seed proteome is highly dynamic, depending on the species variability and environmental influence, we analyzed the proteomic profiles of one transgenic maize variety (event MON 810) in two subsequent generations (T05 and T06) with their respective isogenic controls (WT05 and WT06). Thus, by comparing the proteomic profiles of WT05 with WT06 we could determine the environmental effects, while the comparison between WT06 and T06 seeds from plants grown under controlled conditions enabled us to investigate the effects of DNA manipulation. Finally, by comparison of T05 with T06 seed proteomes, it was possible to get some indications about similarities and differences between the adaptations of transgenic and isogenic plants to the same strictly controlled growth environment. Approximately 100 total proteins resulted differentially modulated in the expression level as a consequence of the environmental influence (WT06 vs WT05), whereas 43 proteins resulted up- or down-regulated in transgenic seeds with respect to their controls (T06 vs WT06), which could be specifically related to the insertion of a single gene into a maize genome by particle bombardment. Transgenic seeds responded differentially to the same environment as compared to their respective isogenic controls, as a result of the genome rearrangement derived from gene insertion. To conclude, an exhaustive differential proteomic analysis allows to determine similarities and differences between traditional food and new products (substantial equivalence), and a case-by-case assessment of the new food should be carried out in order to have a wide knowledge of its features.     

A complementary peptide vaccine that induces T cell anergy and prevents experimental allergic neuritis in Lewis rats.

We have developed and described a new method of altering T cell-mediated autoimmune diseases by immunization with the complementary peptide against T cell epitopes. The complementary peptide (denoted NAE 07-06) to the bovine P2 protein, residues 60-70 (denoted EAN 60-70), was tested in the Lewis rat model of experimental allergic neuritis (EAN). Immunization with NAE 07-06 induced polyclonal and monoclonal Abs that inhibited the proliferation of the P2-specific T cell line, stimulated with EAN 60-70, and recognized Vbeta, but not Valpha, of TCRs. Proliferation of T cells treated with anti-NAE 07-06 Abs could be partially restored by treatment with rIL-2, in accordance with an anergy model. A homologous sequence was found between NAE 07-06 and the VDJ junction of the TCR beta-chain from an EAN 60-70-specific T cell line. Rats preimmunized with NAE 07-06 in vivo before EAN induction showed less disease severity clinically and histologically. These data suggest a new therapeutic approach for T cell-mediated autoimmune disorders through the induction of anti-TCR Abs with complementary peptide Ags.     

Arsukibacterium ikkense gen. nov., sp. nov, a novel alkaliphilic, enzyme-producing gamma-Proteobacterium isolated from a cold and alkaline environment in Greenland

A novel aerobic, Gram-negative, non-pigmented bacterium, GCM72(T), was isolated from the alkaline, low-saline ikaite columns in the Ikka Fjord, SW Greenland. Strain GCM72(T) is a motile, non-pigmented, amylase- and protease-producing, oxidase-positive, and catalase-negative bacterium, showing optimal growth at pH 9.2-10.0, at 15 degrees C, and at 3% (w/v) NaCl. Major fatty acids were C(12:0) 3-OH (12.2+/-0.1%), C(16:00) (18.0+/-0.1%), C(18:1)omega7c (10.7+/-0.5%), and summed feature 3 comprising C(16:1)omega7c and/or iso-C(15:0) 2-OH (36.3+/-0.7%). Phylogenetic analysis based on 16S rRNA gene sequences showed that isolate GCM72(T) was most closely related to Rheinheimera baltica and Alishewanella fetalis of the gamma-Proteobacteria with a 93% sequence similarity to both. The G+C content of DNA isolated from GCM72(T) was 49.9mol% and DNA-DNA hybridization between GCM72T and R. baltica was 9.5%. Fatty acid analysis and G+C content supports a relationship primarily to R. baltica, but several different features, such as a negative catalase-response and optimal growth at low temperature and high pH, together with the large phylogenetic distance and low DNA similarity to its closest relatives, lead us to propose a new genus, Arsukibacterium, gen. nov., with the new species Arsukibacterium ikkense sp. nov. (type strain is GCM72(T)).     

Spermatogenic effects of male-fertile translocations in the mouse

Four male-fertile translocations, T(2;4)13H, T(2.8)26H, T(7;18)50H and T(1;13)70H were crossed to the inbred strains CBA/H and C57BL/6J. F1 heterozygotes were compared with wild-type litter-mates for signs of spermatogenic impairment, in view of previous reports that the C57BL strain had this effect in the T(14;15)6Ca translocation. There was a general tendency for body-weights to be slightly reduced in translocation carriers vs. wild-type. Mean testis weights were significantly reduced on the C57BL background with all four translocations as compared to wild-type, but also significantly increased in T26H on CBA. Sperm counts were also reduced on the C57BL background in T13H, T50H and T70H (significantly so in the last two) but were significantly increased in T13H on a CBA background. Only in T50H did the frequency of sperm-head abnormalities show any marked change in the translocation heterozygotes, being approximately doubled with both CBA and C57BL backgrounds although still remaining at a low level. It was concluded that the deleterious effects of C57BL strains on spermatogenesis in translocation heterozygotes were not confined to T6Ca but were probably widespread. Some inconclusive evidence suggested that this might be because some genetic factors associated with C57BL tended to reduce chiasma frequencies in translocation heterozygotes.     

Enhancement of somatic embryogenesis and plant regeneration in Japanese red pine (Pinus densiflora)

The highest proliferation rate (9.8-fold) of embryogenic suspensor mass (ESM) was obtained from half-strength Litvay (½LM, Litvay et al. 1985) medium supplemented with 3.42 mM l-glutamine, while the lowest rate was noted when 0.84 mM l-glutamine (0.6-fold) was added to the medium. The highest growth ratio with brassinolide (BL) was observed for 1.0 μM (2.3-fold, line 05-21) and 0.05 μM (2.9-fold, line 06-22). However, in the ESM lines 05-21 and 06-22, high ESM growth rates (2.3-fold, line 05-21 and 2.1-fold, line 06-3) were seen without BL when compared with 1.0 μM (05-21) or 0.05 μM (06-22). BL-supplemented medium has been shown to have diverse, genotype-specific effects on the degree of ESM proliferation. For somatic embryo maturation with 0.05 % activated charcoal (AC), the highest number (798 g^?1 FW) of cotyledonary somatic embryos (line 06-29) were obtained on a maturation medium supplemented with AC. The influence of light-emitting diode (LED) sources on the germination of somatic embryos (four genotypes) in this species was studied and was strongly genotype-dependent. Germination of somatic embryos from ESM line 05-3 was strongly inhibited by fluorescent, and red + blue light, while lines 05-12, 05-29, and 05-37 showed a similar germination frequency for the five LED sources, where red light most stimulated somatic embryo germination.     

H-2 antigen expression and sensitivity of BL6 melanoma cells to natural killer cell cytotoxicity

The sensitivity of H-2b-high and H-2b-low variants of BL6 melanoma to the cytotoxic action of NK and lymphokine-activated killer cells was investigated. BL6 mouse melanoma cells lack detectable H-2Kb and had low levels of expression of H-2Db Ag. The BL6T2 variant cells, obtained after treatment of BL6 cells with mutagen N-methyl-N-nitro-N'-nitro-soguanidine, had relatively high levels of expression of class I H-2b Ag. Poly(I:C)-stimulated spleen cells of nude mice were highly cytotoxic for BL6T2, whereas H-2b-low BL6 cells were less sensitive to NK activity in an 18-h 51Cr-release assay. Similar results were obtained after 4-h incubation of radio-labeled tumor cells with IL-2-activated effector cells. In contrast, both lines were equally sensitive to lysis by purified granules derived from rat large granular lymphocytes (LGL) or by macrophages. By using various clones selected from BL6 or BL6T2 cells, it was found that BL6 or BL6T2 clones with low H-2b Ag expression were less sensitive to lysis by NK cells than H-2b-high clones. After IFN treatment of either BL6 or BL6T2, the target cells became more resistant to lysis by either NK cells or by purified LGL granules. IFN-treated BL6 cells had substantially increased expression of H-2b Ag and in this respect became similar to untreated BL6T2. However, IFN-treated BL6 cells were more resistant than BL6T2 cells to lysis by NK cells and LGL granules, suggesting that augmentation of H-2b Ag expression and NK resistance could be two independent IFN-induced effects. With a cold target inhibition assay, it was found that BL6T2 or its H-2 positive clones were highly competitive and inhibited the cytotoxic activity of NK and lymphokine-activated killer cells against radiolabeled YAC-1 and BL6T2, whereas BL6 cells or H-2-negative clones of BL6T2 and BL6 lines showed poor competitive ability. Thus, our data indicate that the NK resistance of H-2-low BL6 cells may be due to a paucity of NK recognizable determinants. N-Methyl-N-nitro-N'-nitroguanidine treatment of BL6 melanoma cells was associated with an increase in class I H-2b Ag expression and NK sensitivity, suggesting the involvement of class I MHC Ag in the sensitivity of tumor cells to NK cell-mediated cytotoxicity.     

Visualization, fate, and pathogenicity of antigen-specific CD8+ T cells in the graft-versus-host reaction.

To follow the fate of alloreactive T cell effectors in graft-vs-host disease, Ld-specific CD8+ T cells from C57BL/6 2C TCR-transgenic donors were transplanted into sublethally irradiated (750 cGy) Ld+ or Ld- recipients. In Ld- C57BL/6 or (BALB/c-dm2 x C57BL/6)F1 recipients, naive 2C T cells engrafted and survived long term, but did not acquire effector function. In Ld+ (BALB/c x C57BL/6)F1 recipients, 2C T cells engrafted, expanded, became cytolytic, destroyed host B cells and double-positive thymocytes, and later disappeared. Despite marked damage to lymphoid and hemopoietic cells by 2C T cells, no significant pathology was detected in other organs, and recipients survived. Ld+ (BALB/c x C57BL/6)F1 recipients died when LPS/endotoxin was administered on day 7 after cell transfer, while Ld- (BALB/c-dm2 x C57BL/6)F1 recipients survived. Our findings show that under certain conditions, a CD8+ T cell population recognizing an extremely limited repertoire of Ags can initiate graft-vs-host disease.     

Discovery of a unique strain-specific idiotypic antigenic determinant of mouse T-lymphocyte antigen-recognizing receptors

The cellular radioimmunoassay using rabbit anti-CBA-anti-C57BL/6 antiidiotypic serum was performed to detect a private strain-specific antigenic idiotypic determinant (s) on activated CBA-anti-C57BL/6 T cells. This private idiotype was not expressed on other activated T cells under study: CBA-anti-BALB/C; C3H-anti-C57BL/6; AKR-anti-C57BL/6; A/Sn-anti-C57BL/6; BALB/C-anti-C57BL/6; DBA/2-anti-C57BL/6. It is suggested that gene (s) coding the private strain-specific idiotypic antigenic determinant of the antigen-recognition receptor of T cells is not localized on the H-2 complex of mice.     

Identification of ‘Amigo’ and ‘Kavkaz’ translocations in Ohio soft red winter wheats (Triticum aestivum L.)

One cultivar (GR876) and two advanced Ohio soft red winter wheat lines (OH413 and OH414), with Kavkaz in their pedigrees, were examined for the presence of the Kavkaz, 1RS/1BL rye/wheat chromosome translocation. Another advanced line (OH416), with Amigo in its pedigree, was examined for the presence of the Amigo, 1RS/1AL translocation. Only two satellited chromosomes were observed in most mitotic root-tip cells from GR876, OH413, and OH414, compared to four in most cells from OH416. Heteromorphic bivalents were observed in most PMCs from hybrids produced by crossing GR876, OH413, and OH414 as females to Chinese Spring. No heteromorphic bivalents were observed in PMCs from OH416 x Chinese Spring hybrids. When GR876 and the Ohio lines were hybridized with Chinese Spring dimonotelosomic-1B, telosomic trivalents, consisting of the short- and longarm telosomes paired with chromosome 1B, were only observed in PMCs from 43-chromosome hybrids involving OH416. The long-arm telosome paired with the translocation chromosome, while the short-arm telosome remained unpaired in all other 43-chromosome hybrids. Separation of gliadin proteins from GR876 and the Ohio lines by PAGE revealed that secalin bands for GR876, OH413, and OH414, migrated similarly to the secalins for Kavkaz. Bands for OH416, identified as possible secalins, migrated similarly to those for Amigo. Cultivar GR876 and advanced Ohio soft red winter wheat lines OH413 and OH414 carry the Kavkaz translocation, while OH416 carries the Amigo translocation.Communicated by K. Tsunewaki     

Interaction of thymus lymphocytes with histoincompatible cells. IV. Mixed lymphocyte reactions of activated thymus lymphocytes

CS7BL-activated CBA T cells (T.TDL) were obtained by thoracic duct cannulation of (CBA × C57BL)F₁ mice 4 days after heavy irradiation and injection of CBA thymus cells. T.TDL behaved differently from the TDL of normal CBA mice in unidirectional mixed lymphocyte culture in a number of respects: (a) the response of T.TDL was directed specifically against C57BL antigens, whereas normal TDL responded to both C57BL and BALB/c antigens; (b) the response of T.TDL was rapid but transient compared to that of TDL; (c) whereas only approximately 3% of TDL synthesized DNA specifically in response to C57BL antigens, as many as 25% of C57BL-activated T.TDL responded to these antigens. Evidence is presented which suggests that the T.TDL have a very limited capacity to proliferate. Most of the cells which responded to antigen synthesized DNA without subsequently entering mitosis.     

Chromosome composition of wheat-rye lines and the influence of rye chromosomes on disease resistance and agronomic traits

Identification of the chromosomal composition of common wheat lines with rye chromosomes was carried out using genomic in situ hybridization and 1RS- and 5P-specific PCR markers. It was demonstrated that wheat chromosomes 5A or 5D were substituted by rye chromosome 5R in the wheat-rye lines. It was established that one of the lines with complex disease resistance contained rye chromosome 5R and T1RS.1BL, while another line was found to contain, in addition to T1RS.1BL, a new Robertsonian translocation, T5AS.5RL. Substitution of the wheat chromosome 5A with the dominant Vrn-A1 gene for the Onokhoiskaya rye chromosome 5R led to lengthening of the germination-heading period or to a change in the type of development. A negative influence of T1RS.1BL on SDS sedimentation volume and grain hardness was demonstrated, along with a positive effect of the combination of T1RS.1BL and 5R(5D) substitution on grain protein content. Quantitative traits of the 5R(5A) and 5R(5D) substitution lines were at the level of recipient cultivars. A line with two translocations, T1RS.1BL + T5AS.5Rl, appeared to be more productive as compared to the line carrying T1RS.1BL in combination with the 5R(5D) substitution.     

海南热带雨林腐木上两个无性型子囊菌酵母新种

用含木糖为唯一碳源和含葡萄糖及7.6%乙醇的两种富集培养基对采自海南热带雨林腐木样品中的酵母菌进行了分离培养。对分离出的酵母菌株进行的分子分类学研究表明,其中两株酵母菌X2WZ07-4和G2WZ06-1代表两个无性型子囊菌酵母新种。大亚基(26S)rRNA基因D1/D2域序列分析显示,与X2WZ07-4和G2WZ06-1亲缘关系最近的已知种分别为Candida cylindracea和C.llanquihuensis。在D1/D2域,X2WZ07-4与C.cylindracea模式菌株的碱基序列差异为2.5%;G2WZ06-1与C.llanquihuensis模式菌株的序列差异为3.9%,均远大于酵母菌种间在此区域的序列差异(～1%)。这两个新种分别被命名为拟柱形假丝酵母Candida pseudocylindracea sp.nov.(模式菌株:X2WZ07-4T=AS2.3788T=CBS10854T)和五指山假丝酵母Candida wuzhishanensis sp.nov.(模式菌株:G2WZ06-1T=AS2.3784T=CBS10850T)。