Culture-based study of the faecal microbiome in two adult female subjects revealed the presence of two obligately anaerobic, non-spore-forming, rod-shaped, non-motile, Gram-negative bacterial strains that represent novel species. The first strain, designated 627^T, was a fastidious, slow-growing, indole-positive bacterium with a non-fermentative type of metabolism. The strain was characterized by the production of acetic and succinic acids as metabolic end products, the prevalence of iso-C_15 : 0fatty acid and the presence of menaquinones MK-10 and MK-11. The DNA G+C content was found to be 56.6 mol%. The second strain, designated 177^T, was capable of fermenting a rich collection of carbohydrate substrates, producing acetic acid as a terminal product. The strain was indole-negative and resistant to bile. The major cellular fatty acids were iso-C_15 : 0and anteiso-C_15 : 0(in a 1 : 1 ratio) and the predominant menaquinone was MK-11. The DNA G+C content was 37.8 mol%. A phylogenomic analysis of the draft genomes of strains 627^Tand 177^Tplaced these bacteria in the generaAlistipes(familyRikenellaceae) andCoprobacter(familyPorphyromonadaceae), respectively.On the basis of the phenotypic and genotypic properties of strains 627^Tand 177^T, we conclude that these strains from human faeces represent two novel bacterial species, for which the namesAlistipes inopssp. nov. (type strain 627^T = DSM 28863^T = VKM B-2859^T) andCoprobacter secundussp. nov. (type strain 177^T = DSM 28864^T = VKM B-2857^T) are proposed.

Two novel strains of Gram-negative staining, rod-shaped, obligately anaerobic, non-sporeforming, non-motile bacteria were isolated from the feces of healthy human subjects. The strains designated as 585-1T and 668 are characterized by mesophilic fermentative metabolism, production of D-lactic, succinic, and acetic acids as end products of D-glucose fermentation, prevalence of C18:1ω9, C18:1ω9a, C16:0, and C16:1ω7-сis fatty acids, presence of glycine, glutamic acid, lysine, alanine, and aspartic acid in petidoglycan peptide moiety and lack of respiratory quinones. Whole genome sequencing revealed the DNA G+C content was 56.4-56.6 mol%. Complete 16S rRNA gene sequences shared 91.7/91.6% identity with Anaerofilum pentosovorans FaeT, 91.3/91.2% with Gemmiger formicilis ATCC 27749T, and 88.9/88.8% with Faecalibacterium prausnitzii ATCC 27768T. On the basis of chemotaxonomic and genomic properties it was concluded that the strains represent a new species in a new genus within the family Ruminococcaceae, for which the name Ruthenibacterium lactatiformans gen. nov., sp. nov. is proposed. The type strain is 585-1T (= DSM 100348T, = VKM B-2901T).

Th1 lymphocytes are considered the main mediators of protection against tuberculosis (TB); however, their phenotypic characteristics and relationship with Th17 and Th1Th17 populations during TB are poorly understood. We have analyzed Th1, Th17, and Th1Th17 lymphocytes in the blood and pulmonary lesions of TB patients. The populations were identified based on the production of IFN-γ and/or IL-17 and the coexpression of CXCR3 (X3) and CCR6 (R6). In the blood, IL-17⁺ and IFN-γ⁺IL-17⁺ lymphocytes were barely detectable (median, <0.01% of CD4⁺ lymphocytes), whereas IFN-γ⁺ lymphocytes predominated (median, 0.45%). Most IFN-γ⁺ lymphocytes (52%) were X3⁺R6⁺, suggesting their “nonclassical” (ex-Th17) nature. In the lungs, IL-17⁺ and IFN-γ⁺IL-17⁺ lymphocytes were more frequent (0.3%, p < 0.005), yet IFN-γ⁺ cells predominated (11%). Phenotypically, lung CD4⁺ cells were X3^+/loR6⁻. The degree of differentiation of blood effector CD4⁺ lymphocytes (evaluated based on CD62L/CD27/CD28 coexpression) increased as follows: X3⁺R6⁺ < X3⁺R6⁻ < X3⁻R6⁻, with X3⁻R6⁻ cells being largely terminally differentiated CD62L⁻CD27⁻CD28⁻ cells. Lung CD4⁺ lymphocytes were highly differentiated, recalling blood X3^+/−R6⁻ populations. Following in vitro stimulation with anti-CD3/anti-CD28 Abs, X3⁺R6⁺CD4⁺ lymphocytes converted into X3⁺R6⁻ and X3⁻R6⁻ cells. The results demonstrate that, during active TB, Th1 lymphocytes predominate in blood and lungs, document differences in X3/R6 expression by blood and lung CD4⁺ cells, and link the pattern of X3/R6 expression with the degree of cell differentiation. These findings add to the understanding of immune mechanisms operating during TB and are relevant for the development of better strategies to control it.

This work presents a detailed evaluation of the performance of density functional theory (DFT) for the prediction of zero-field splittings (ZFSs) in high-spin nitrenes. A number of well experimentally characterized triplet mononitrenes, quartet nitrenoradicals, quintet dinitrenes, and septet trinitrenes have been considered. Several DFT-based approaches for the prediction of ZFSs have been compared. It is shown that the unrestricted Kohn–Sham and the Pederson–Khanna approaches are the most successful for the estimation of the direct spin-spin (SS) interaction and the spin-orbit coupling (SOC) parts, respectively, to the final ZFS parameters. The most accurate theoretical predictions (within 10%) are achieved by using the PBE density functional in combination with the DZ, EPR-II, and TZV basis sets. For high-spin nitrenes constituted from light atoms, the contribution of the SOC part to ZFS parameters is quite small (7%–12%). By contrast, for chlorine-substituted septet trinitrenes, the contribution of the SOC part is small only to D value but, in the case of E value, it is as large as the SS part and has opposite sign. Due to this partial cancellation of two different contributions, SS and SOC, the resulting values of E in heavy molecules are almost two times smaller than those predicted by analysis of the widely used semiempirical one-center spin-spin interaction model. The decomposition of DSS into n-center (n=1–4) interactions shows that the major contribution to DSS results from the one-center spin-spin interactions. This fact indicates that the semiempirical SS interaction model accurately predicts the ZFS parameters for all types of high-spin nitrenes with total spin S=2 and 3, if their molecules are constructed from the first-row atoms.