Effect of BCG on Friend Disease Virus in Mice

NONSPECIFIC stimulation of the reticulo-endothelial system and resistance against tumorigenesis are produced by injection of attenuated Mycobacterium bovis (BCG). Biozzi et al.¹ demonstrated resistance to Ehrlich's ascites tumour in BCG-immune mice and Halpern et al.² showed that such immunization effected control of T-8 tumours in rats. Further, at least partial control of other tumours^3–6 has been effected by this procedure in mice and hamsters.     

Anti-θ Serum-indueed Suppression of the Cellular Transfer of Tumour-specific Immunity to a Syngeneic Plasma Cell Tumour

IT has been well documented that tumour-bearing mice can become resistant to their own tumours, especially with chemically induced fibrosarcomas^1–3 and the importance of cell-mediated immune responses rather than humoral antibody in the resistance to tumour transplants has been emphasized^3,4, although the exact mechanism of tumour cell destruction remains ill-defined. Studies in mice^5,6, using allogeneic tumour cells, have demonstrated that thymus-derived (T) lymphocytes are essential for the killing of tumour cells. In addition, using an in vitro method of immunization against histocompatibility antigens, tumour cell destruction either in vitro¹ or in vivo⁸ was shown to be due to T cells alone. In all of these latter studies, however, it is the strong H-2 histocompatibility antigens that are inducing the immune response and not the tumour-specific transplantation antigens (TSTA). We describe here a specific anti-TSTA response to a murine plasma cell tumour which can be transferred with lymphoid cells and which can be shown to involve the essential participation of T cells.     

Stereochemistry of Intercalation: Interaction of Daunomycin with DNA

DAUNOMYCIN^1–3, a glycosidic anthracycline antibiotic from Streptomyces peucetius⁴, is being used in the treatment of acute leukaemia and solid tumours in man^5,6. The biological activity seems to be due to complex formation with the DNA of deoxyribonucleoprotein⁴. In vivo, daunomycin inhibits both RNA and DNA synthesis^7,8 and, in vitro, DNA-dependent RNA polymerase and DNA polymerase^7–9.     

Role of PKCtheta in macrophage-mediated immune response to <Emphasis Type="Italic">Salmonella typhimurium</Emphasis> infection in mice

Background

The serine/threonine protein kinase C (PKC) theta has been firmly implicated in T cell-mediated immunity. Because its role in macrophages has remained undefined, we employed PKCtheta-deficient (PKCtheta ^?/?) mice in order to investigate if PKCtheta plays a role in macrophage-mediated immune responses during bacterial infections.

Results

Our results demonstrate that PKCtheta plays an important role in host defense against the Gram-negative, intracellular bacterium Salmonella typhimurium, as reflected both by markedly decreased survival and a significantly enhanced number of bacteria in spleen and liver of PKCtheta ^?/? mice, when compared to wild-type mice. Of note, albeit macrophages do not express detectable PKCtheta, PKCtheta mRNA expression was found to be profoundly upregulated during the first hours of lipopolysaccharide (LPS)/interferon-gamma (IFNgamma)-, but not IL-4-mediated cell polarization conditions in vitro. Mechanistically, despite expressing normal levels of classically activated macrophage (CAM) markers, PKCtheta-deficient CAMs expressed significantly higher levels of the anti-inflammatory cytokine IL-10 in vivo and in vitro when challenged with S. typhimurium or LPS/IFNgamma. Neutralization of IL-10 recovered immune control to S. typhimurium infection in PKCtheta-deficient macrophages.

Conclusions

Taken together, our data provide genetic evidence that PKCtheta promotes a potent pro-inflammatory CAM phenotype that is instrumental to mounting protective anti-bacterial immunity. Mechanistically, PKCtheta exerts a host-protective role against S. typhimurium infection, and acts as an essential link between TLR4/IFNgammaR signaling and selective suppression of the anti-inflammatory cytokine IL-10 at the onset of CAM differentiation in the course of a bacterial infection.

     

The role of the P2X7 receptor in murine cutaneous leishmaniasis: aspects of inflammation and parasite control

Leishmania amazonensis is the etiological agent of diffuse cutaneous leishmaniasis. The immunopathology of leishmaniasis caused by L. amazonensis infection is dependent on the pathogenic role of effector CD4⁺ T cells. Purinergic signalling has been implicated in resistance to infection by different intracellular parasites. In this study, we evaluated the role of the P2X7 receptor in modulating the immune response and susceptibility to infection by L. amazonensis. We found that P2X7-deficient mice are more susceptible to L. amazonensis infection than wild-type (WT) mice. P2X7 deletion resulted in increased lesion size and parasite load. Our histological analysis showed an increase in cell infiltration in infected footpads of P2X7-deficient mice. Analysis of the cytokine profile in footpad homogenates showed increased levels of IFN-γ and decreased TGF-β production in P2X7-deficient mice, suggesting an exaggerated pro-inflammatory response. In addition, we observed that CD4⁺ and CD8⁺ T cells from infected P2X7-deficient mice exhibit a higher proliferative capacity than infected WT mice. These data suggest that P2X7 receptor plays a key role in parasite control by regulating T effector cells and inflammation during L. amazonensis infection.     

Improvement of seedling and panicle blast resistance in <Emphasis Type="Italic">Xian</Emphasis> rice varieties following <Emphasis Type="Italic">Pish</Emphasis> introgression

Rice blast is a serious disease caused by the filamentous ascomycetous fungus Magnaporthe oryzae. Incorporating disease resistance genes in rice varieties and characterizing the distribution of M. oryzae isolates form the foundation for enhancing rice blast resistance. In this study, the blast resistance gene Pish was observed to be differentially distributed in the genomes of rice sub-species. Specifically, Pish was present in 80.5% of Geng varieties, but in only 2.3% of Xian varieties. Moreover, Pish conferred resistance against only 23.5% of the M. oryzae isolates from the Geng-planting regions, but against up to 63.2% of the isolates from the Xian-planting regions. Thus, Pish may be an elite resistance gene for improving rice blast resistance in Xian varieties. Therefore, near-isogenic lines (NILs) with Pish and the polygene pyramid lines (PPLs) PPL^Pish/Pi1, PPL^Pish/Pi54, and PPL^Pish/Pi33 in the Xian background Yangdao 6 were generated using a molecular marker-assisted selection method. The results suggested that (1) Pish significantly improved rice blast resistance in Xian varieties, which exhibited considerably improved seedling and panicle blast resistance after Pish was introduced; (2) PPLs with Pish were more effective than the NILs with Pish regarding seedling and panicle blast resistance; (3) the PPL seedling and panicle blast resistance was improved by the complementary and overlapping effects of different resistance genes; and (4) the stability of NIL and PPL resistance varied under different environmental conditions, with only PPL^Pish/Pi54 exhibiting highly stable resistance in three natural disease nurseries (Jianyang, Jinggangshan, and Huangshan). This study provides new blast resistance germplasm resources and describes a novel molecular strategy for enhancing rice blast resistance.     

Intralesional uridine-5′-triphosphate (UTP) treatment induced resistance to <Emphasis Type="Italic">Leishmania amazonensis</Emphasis> infection by boosting Th<Subscript>1</Subscript> immune responses and reactive oxygen species production

Leishmania amazonensis is the etiologic agent of cutaneous leishmaniasis, an immune-driven disease causing a range of clinical symptoms. Infections caused by L. amazonensis suppress the activation and function of immune cells, including macrophages, dendritic cells, and CD4⁺ T cells. In this study, we analyzed the course of infection as well as the leishmanicidal effect of intralesional UTP treatment in L. amazonensis-infected BALB/c mice. We found that UTP treatment reduced the parasitic load in both footpad and lymph node sites of infection. UTP also boosted Th1 immune responses, increasing CD4⁺ T cell recruitment and production of IFN-γ, IL-1β, IL-12, and TNF-α. In addition, the role of UTP during innate immune response against L. amazonensis was evaluated using the air pouch model. We observed that UTP augmented neutrophil chemoattraction and activated microbicidal mechanisms, including ROS production. In conclusion, our data suggested an important role for this physiological nucleotide in controlling L. amazonensis infection, and its possible use as a therapeutic agent for shifting immune responses to Th1 and increasing host resistance against L. amazonensis infection.     

Activity and Properties of Macrophage Migration Inhibitory Factor produced by Mixed Lymphocyte Cultures

THE macrophage migration test is an in vitro demonstration of delayed hypersensitivity. Supernatant fluids of sensitive lymphocytes cultured for 24 h in the presence of specific antigen contain migration inhibitory factor (MIF) that arrests the migration of macrophages of unsensitized animals in vitro¹,². In vivo, it induces delayed skin reactions³. The use of the macrophage migration test, based on differences of transplantation antigens in donor and recipient, to show histocompatibility has been suggested⁴. The test was also recommended as an indicator of immunological reactivity after organ transplantation, to demonstrate impending rejection⁵. It can demonstrate homograft sensitivity, for migration of peritoneal exudate cells (containing lymphocytes and macrophages) of CBA mice previously sensitized by grafts from A/Jax donors was inhibited when they were mixed with peritoneal exudate cells of the donor strain. However, histocompatibility was not demonstrated, for mixtures of peritoneal exudate cells of ungrafted CBA mice and A/Jax mice migrated regularly during the 24 h test⁶.     

Friend retrovirus drives cytotoxic effectors through Toll-like receptor 3

Background

Pathogen recognition drives host defense towards viral infections. Specific groups rather than single members of the protein family of pattern recognition receptors (PRRs) such as membrane spanning Toll-like receptors (TLRs) and cytosolic helicases might mediate sensing of replication intermediates of a specific virus species. TLR7 mediates host sensing of retroviruses and could significantly influence retrovirus-specific antibody responses. However, the origin of efficient cell-mediated immunity towards retroviruses is unknown. Double-stranded RNA intermediates produced during retroviral replication are good candidates for immune stimulatory viral products. Thus, we considered TLR3 as primer of cell-mediated immunity against retroviruses in vivo.

Results

Infection of mice deficient in TLR3 (TLR3^?/?) with Friend retrovirus (FV) complex revealed higher viral loads during acute retroviral infection compared to wild type mice. TLR3^?/? mice exhibited significantly lower expression levels of type I interferons (IFNs) and IFN-stimulated genes like Pkr or Ifi44, as well as reduced numbers of activated myeloid dendritic cells (DCs) (CD86⁺ and MHC-II⁺). DCs generated from FV-infected TLR3^?/? mice were less capable of priming virus-specific CD8⁺ T cell proliferation. Moreover, cytotoxicity of natural killer (NK) cells as well as CD8⁺ T cells were reduced in vitro and in vivo, respectively, in FV-infected TLR3^-/- mice.

Conclusions

TLR3 mediates antiretroviral cytotoxic NK cell and CD8⁺ T cell activity in vivo. Our findings qualify TLR3 as target of immune therapy against retroviral infections.

     

Cytotoxic Effects of Peritoneal Neutrophils on a Syngeneic Rat Tumour

SPECIFIC immunological phenomena directed against tumour cells which influence their behaviour and growth in vivo and in vitro have been clearly established^1,2. In contrast, nonspecific defences against cancer have received little attention, although such mechanisms are important in preventing microbial infection³. Recently there have been indications that such non-specific defences against cancer cells exist but the underlying processes are undefined. Increases in reticuloendothelial activity frequently occur during tumour growth⁴; stimulants of non-specific defences, such as the BCG strain of Mycobacterium tuberculosis and Corynebacterium parvum, inhibit tumour growth^5,6; and induction of inflammation by vaccinia⁷ or dichloronitrobenzene⁸ in skin overlying a tumour may induce regression. Most attempts to analyse these phenomena implicate unsensitized lymphocytes⁹ and macrophages¹⁰, although the cytotoxic properties of these cells compared with specifically sensitized cells are low¹¹. Little has been written about the possible role of neutrophils, although their presence in tumours is not infrequent¹². The probable role of neutrophils in contributing to the cytotoxic activity in vitro of leucocyte populations towards various allogeneic and syngeneic cell types has been discussed¹³ and Bubenik et al.¹⁴ have postulated an action of neutrophils against human bladder tumours. Recently Godleski et al.¹⁵ showed movement of neutrophils towards cells of Walker carcinosarcoma 256 growing on chick chlorioallantoic membranes or cover-slips, followed by contact and damage to tumour cell membranes. This article describes preliminary studies demonstrating in vitro and in vivo a significant cytotoxic effect of rat peritoneal neutrophils against a syngeneic ascites tumour, WBP1(A), < 10 cells of which will produce a tumour^16,17.     

Programmed cell death in the cyanobacterium <Emphasis Type="Italic">Microcystis aeruginosa</Emphasis> induced by allelopathic effect of submerged macrophyte <Emphasis Type="Italic">Myriophyllum spicatum</Emphasis> in co-culture system

This investigation demonstrates that programmed cell death (PCD) in a cyanobacterium, Microcystis aeruginosa, resulting from allelopathic stress induced by a submerged macrophyte, Myriophyllum spicatum, in a co-culture system. The hallmarks of PCD, caspase-3-like protease activity, DNA fragmentation, and destruction of cell ultrastructure, as well as intracellular PCD signaling radicals, reactive oxygen species (ROS), and nitric oxide (NO), were measured in M. aeruginosa cells co-cultured with M. spicatum for 7 days. The results showed a dose–response relationship between M. spicatum biomass and M. aeruginosa mortality. A caspase-3-like protease was activated and elevated from day 3. Thylakoid disintegration, cytoplasmic vacuolation, and fuzzy nuclear zone were observed by transmission electron microscopy, and distinct DNA fragmentation was detected in M. aeruginosa cells at a M. spicatum biomass of 6.0 g fresh weight (FW) L^?1 during the 7 days. Allelochemicals of total phenolic compounds (TPCs) were determined in co-culture water, and the concentration increased with increasing of M. spicatum biomass and co-culture time. Compared with the level of ROS production in the control group, a significant overproduction of ROS was detected in M. aeruginosa cells in the treatment group, and this was positively correlated with TPC concentration. Furthermore, the level of intracellular NO increased with the percent mortality of M. aeruginosa. The results indicated that a PCD pathway was induced in the cyanobacterium M. aeruginosa when co-cultured with the submerged macrophyte M. spicatum.     

Gene <Emphasis Type="Italic">angora</Emphasis> as a modifier of the <Emphasis Type="Italic">hairless</Emphasis> gene in mouse

The interaction between mouse angora-Y (Fgf5 ^go-Y) and hairless (hr) genes have been studied. Homozygous mutant gene Fgf5 ^go-Y increases length of all hair types, while homozygotes for the h gene lose hair completely starting on day 14 after birth. We obtained mice with genotypes +/+ hr/hr F₂, +/Fgf5 ^go-Y hr/hr and Fgf5 ^go-Y/Fgf5 ^go-Y hr/hr. Both +/Fgf5 ^go-Y hr/hr and +/+ hr/hr mice began to loose hair from their heads on day 14. This further extended on the whole body. On day 21 the mice were completely deprived of hair. Therefore a single dose of gene Fgf5 ^go-Y does not modify alopecia in mice homozygous for hr. However in double homozygotes Fgf5 ^go-Y/Fgf5 ^go-Y hr/hr alopecia started 4 days later, namely on day 18. It usually finished 10–12 days after detection of first bald patches. On days 28–30 double homozygotes lose coat completely. Hair loss in double homozygous mice was 1.5-fold slower than in +/+ hr/hr mice. This resulted from a significant extension of anagen phase induced by a mutant homozygous gene Fgf5 ^go-Y in morphogenesis of the hair follicle. The hr gene was expressed at the transmission phase from anagen to catagen. Our data shows that the angora gene is a modifier of the hairless gene and this results in a strong repression of alopecia progression in double homozygous mice compared to +/+ hr/hr animals.     

Molecular mapping of a new temperature-sensitive gene <Emphasis Type="Italic">LrZH22</Emphasis> for leaf rust resistance in Chinese wheat cultivar Zhoumai 22

Leaf rust, caused by Puccinia triticina, is one of the most widespread diseases in common wheat globally. The Chinese wheat cultivar Zhoumai 22 is highly resistant to leaf rust at the seedling and adult stages. Seedlings of Zhoumai 22 and 36 lines with known leaf rust resistance genes were inoculated with 13 P. triticina races for gene postulation. The leaf rust response of Zhoumai 22 was different from those of the single gene lines. With the objective of identifying and mapping, the new gene(s) for resistance to leaf rust, F₁, F₂ plants and F_2:3 lines from the cross Zhoumai 22/Chinese Spring were inoculated with Chinese P. triticina race FHDQ at the seedling stage. A single dominant gene, tentatively designated LrZH22, conferred resistance. To identify other possible genes in Zhoumai 22, ten P. triticina races avirulent on Zhoumai 22 were used to inoculate 24 F_2:3 lines. The same gene conferred resistance to all ten avirulent races. A total of 1300 simple sequence repeat (SSR) markers and 36 EST markers on 2BS were used to test the parents, and resistant and susceptible bulks. Resistance gene LrZH22 was mapped in the chromosome bin 2BS1-0.53-0.75 and closely linked to six SSR markers (barc183, barc55, gwm148, gwm410, gwm374 and wmc474) and two EST markers (BF202681 and BE499478) on chromosome arm 2BS. The two closest flanking SSR loci were Xbarc55 and Xgwm374 with genetic distances of 2.4 and 4.8 cM from LrZH22, respectively. Six designated genes (Lr13, Lr16, Lr23, Lr35, Lr48 and Lr73) are located on chromosome arm 2BS. In seedling tests, LrZH22 was temperature sensitive, conferring resistance at high temperatures. The reaction pattern of Zhoumai 22 was different from that of RL 4031 (Lr13), RL 6005 (Lr16) and RL 6012 (Lr23), Lr35 and Lr48 are adult-plant resistance genes, and Lr73 is not sensitive to the temperature. Therefore, LrZH22 is likely to be a new leaf rust resistance gene or allele.     

Apoptotic Endonuclease EndoG Induces Alternative Splicing of Telomerase TERT Catalytic Subunit,Caspase-2, DNase I,and BCL-x in Human,Murine, and Rat CD4<Superscript>+</Superscript> T Lymphocytes

Apoptotic endonuclease EndoG plays a key role in the alternative splicing of mRNA of human TERT telomerase catalytic subunit. The aim of this work was to test the ability of EndoG to induce alternative splicing of mRNA of other genes and in other organisms. To determine new mRNA splice-variants, EndoG overexpression was induced in human, mouse and rat CD4⁺-T-lymphocytes followed by sequencing of total RNA of these cells. Sequencing results showed that besides TERT, EndoG induced alternative splicing of deoxyribonuclease I (DNase I), caspase-2 (Casp-2) and BCL-x. The expression level of EndoG strongly correlated with mRNA splicing-variants of TERT, DNase I, Casp-2, and BCL-x in intact CD4⁺-T cells of healthy donors as well as different lines of mice and rats. EndoG overexpression induced down-regulation of fulllength mRNAs of TERT, DNase I, Casp-2, and BCL-x and up-regulation of their short-length mRNAs. Alternative splicing of studied mRNAs resulted in down-regulation of enzymatic activity of proteins in vitro and in vivo. The results of this work confirm the ability of endonuclease EndoG to induce alternative splicing of several mRNAs in human, mice and rats.     

Regulatory roles of miR-155 and let-7b on the expression of inflammation-related genes in THP-1 cells: effects of fatty acids

The main aim of this investigation was to study the regulatory roles of let-7b and miR-155-3p on the expression of inflammation-associated genes in monocytes, macrophages, and lipopolysaccharide (LPS)-activated macrophages (AcM). A second goal was to analyze the potential modulatory roles of different fatty acids, including oleic, palmitic, eicosapentaenoic (EPA), and docosahexaenoic (DHA), on the expression of these miRNAs in the three cell types. This hypothesis was tested in human acute monocytic leukemia cells (THP-1), which were differentiated into macrophages with 2-O-tetradecanoylphorbol-13-acetate (TPA) and further activated with LPS for 24 h. Monocytes, macrophages, and AcM were transfected with a negative control, or mimics for miR-155-3p and miR-let-7b-5p. The expression of both miRNAs and some proinflammatory genes was analyzed by qRT-PCR. Interestingly, let-7b mimic reduced the expression of IL6 and TNF in monocytes, and SERPINE1 expression in LPS-activated macrophages. However, IL6, TNF, and SERPINE1 were upregulated in macrophages by let-7b mimic. IL6 expression was higher in the three types of cells after transfecting with miR-155-3p mimic. Similarly, expression of SERPINE1 was increased by miR-155-3p mimic in monocytes and macrophages. However, TLR4 was downregulated by miR-155-3p in monocytes and macrophages. Regarding the effects of the different fatty acids, oleic acid increased the expression of let-7b in macrophages and AcM and also increased the expression of miR-155 in monocytes when compared with DHA but not when compared with non-treated cells. Overall, these results suggest anti- and proinflammatory roles of let-7b and miR-155-3p in THP-1 cells, respectively, although these outcomes are strongly dependent on the cell type. Noteworthy, oleic acid might exert beneficial anti-inflammatory effects in immune cells (i.e., non-activated and LPS-activated macrophages) by upregulating the expression of let-7b.     

Nitrification in intertidal sponge <Emphasis Type="Italic">Cinachyrella cavernosa</Emphasis>

The nitrification process in different sections of the sponges remains unresolved, despite several studies on the nitrogen cycling pathways in the tissues of temperate and Arctic sponges. In this study, the abundance, diversity and activity of the associated nitrifying organisms in intracellular, intercellular, extracellular and cortex of a tropical intertidal sponge, Cinachyrella cavernosa, were investigated using most probable number, next-generation sequencing and incubation method, respectively. The nitrification rate and the abundance of nitrifying bacteria showed significant difference among different sections. The nitrification rate in C. cavernosa was 2–12× higher than the reported values in other sponge species from temperate and Arctic regions. Nitrification rate in sponge cortex was 2× higher than in intercellular and extracellular sections. Ammonium and nitrite oxidisers ranged from 10³ to 10⁴ CFU g^?1 in the sponge with a high number of ammonium and nitrite oxidisers in the cortex. Nitrifiers belonging to Nitrosomonas, Nitrospira, Nitrospina, Nitrobacter and Nitrosopumilus were present in different sections of the sponge, with nitrifying archaea dominating the intracellular section and nitrifying bacteria dominating other sections. This study reports for the first time the nitrification inside the sponge cells. The study also suggests that the intertidal sponge, C. cavernosa, harbours metabolically active nitrifiers in different sections of the sponge body with different rates of nitrification. Thus, nitrifiers play an important role in ammonia detoxification within the sponge and also contribute to the nitrogen budget of the coastal ecosystem.     

Evaluation of phylogenetic relationships in the genus <Emphasis Type="Italic">Mus</Emphasis> using <Emphasis Type="Italic">t</Emphasis>-specific microsatellite DNA markers

The t-complex includes a complex system of genes localized in the proximal region of chromosome 17 of house mouse Mus musculus. The results of microsatellite analysis of laboratory stocks of house mice carrying t ¹², t ^w5, t ^w12, and t ^w73 haplotypes and wild mice from natural populations of Russia (Volgograd, Rostov, Saratov oblasts, and Kalmykia), Armenia, Bulgaria, Iran, and Mongolia performed by the PCR method with the use of eight pairs of D17Mit primers (16, 21, 23, 28, 32, 57, 63, 78) are presented. These pairs of primers amplify microsatellite DNA sequences on mouse chromosome 17 in the region from 7.6 to 18.8 cM that correspond to inversions (In (17) 3.4). Each pair of primers recognized three to six variants of nucleotide sequences ranging in size from 90–120 bp (D17Mit 16) to 300–330 bp (D17Mit 57). In most cases, two variants of nucleotide sequences were detected in each individual, i. e., most individuals were heterozygous for the microsatellite loci under study. The highest similarity of the spectra of microsatellite DNA fragments was revealed in laboratory stocks of house mice carrying the t ^w5 and t ^w73 haplotypes. The spectra of animals from the Rostov and Volgograd oblasts appeard to be most similar to them. The microsatellite spectra of individuals from Iran closely resemble the spectrum of an individual from Armenia. It was demonstrated that amplified microsatellite fragments localized in the region of the t-complex can be used to identify representatives of the Mus genus from wild populations.     

<Emphasis Type="Italic">Oceanobacillus gochujangensis</Emphasis> sp. nov., isolated from <Emphasis Type="Italic">gochujang</Emphasis> a traditional Korean fermented food

A Gram-stain-positive, polar flagella-containing, rod-shaped, obligate aerobic, endospore-forming bacterium, strain TK1655^T, was isolated from the traditional Korean food gochujang. The 16S rRNA sequence of strain TK1655^T was a member of the genus Oceanobacillus similar to that of the type strain of Oceanobacillus oncorhynchi subsp. incaldanensis DSM 16557^T (97.2%), O. oncorhynchi subsp. oncorhynchi JCM 12661^T (97.1%), O. locisalsi KCTC 13253^T (97.0%), and O. sojae JCM 15792^T (96.9%). Strain TK1655^T was oxidase and catalase positive. Colonies were circular, smooth, low convex, cream in colour, and measured about 0.5–1.0 mm in diameter. The range for growth was 20–40°C (optimal, 30°C), pH 6.0–10.0 (optimal, 7.0), and 2–16% (w/v) NaCl (optimal, 2%). Additionally, the cells contained meso-DAP, and the predominant isoprenoid quinone was MK-7. The complex polar lipids were consisted of diphosphatidylglycerol (DPG), phosphatidylglycerol (PG), phosphatidylcholine (PC). The major cellular fatty acid components were iso-C_15:0, anteiso-C_15:0, iso-C_16:0, and anteiso-C_17:0, and the DNA G+C content was 40.5%. DNA-DNA relatedness of our novel strain and reference strain O. locisalsi KCTC 13253^T, O. oncorhynchi subsp. incaldanensis DSM 16557^T, O. oncorhynchi subsp. oncorhynchi JCM 12661^T was 45.7, 43.8, and 41.9%. From the results of phenotypic, chemotaxonomic, and phylogenetic analyses of strain TK1655^T, we propose the novel species Oceanobacillus gochujangensis sp. nov. The type strain is TK1655^T (=KCCM 101304^T =KCTC 33014^T =CIP 110582^T =NBRC 109637^T).     

Inflammation in Lafora Disease: Evolution with Disease Progression in Laforin and Malin Knock-out Mouse Models

Lafora progressive myoclonus epilepsy (Lafora disease, LD) is a fatal rare autosomal recessive neurodegenerative disorder characterized by the accumulation of insoluble ubiquitinated polyglucosan inclusions in the cytoplasm of neurons, which is most commonly associated with mutations in two genes: EPM2A, encoding the glucan phosphatase laforin, and EPM2B, encoding the E3-ubiquitin ligase malin. The present study analyzes possible inflammatory responses in the mouse lines Epm2a ^?/? (laforin knock-out) and Epm2b ^?/? (malin knock-out) with disease progression. Increased numbers of reactive astrocytes (expressing the GFAP marker) and microglia (expressing the Iba1 marker) together with increased expression of genes encoding cytokines and mediators of the inflammatory response occur in both mouse lines although with marked genotype differences. C3ar1 and CxCl10 messenger RNAs (mRNAs) are significantly increased in Epm2a ^?/? mice aged 12 months when compared with age-matched controls, whereas C3ar1, C4b, Ccl4, CxCl10, Il1b, Il6, Tnfα, and Il10ra mRNAs are significantly upregulated in Epm2b ^?/? at the same age. This is accompanied by increased protein levels of IL1-β, IL6, TNFα, and Cox2 particularly in Epm2b ^?/? mice. The severity of inflammatory changes correlates with more severe clinical symptoms previously described in Epm2b ^?/? mice. These findings show for the first time increased innate inflammatory responses in a neurodegenerative disease with polyglucosan intraneuronal deposits which increase with disease progression, in a way similar to what is seen in neurodegenerative diseases with abnormal protein aggregates. These findings also point to the possibility of using anti-inflammatory agents to mitigate the degenerative process in LD.