A novel missense variant in HIKESHI: Clinical phenotype, in vitro functional testing, and potential for gene therapy.

A 7-month-old boy presented to our clinic with developmental delay, Magnetic Resonance Imaging (MRI) features of delayed myelination and diffusion restriction, and a homozygous variant of uncertain significance (c.4T>G, p.Phe2Val) in HIKESHI, a gene associated with autosomal-recessive hypomyelinating leukodystrophy 13. We hypothesized that the variant is disease-causing and aimed to rescue the cellular phenotype with vector-mediated gene replacement. HIKESHI mediates heat-induced nuclear accumulation of heat-shock proteins, including HSP70, to protect cells from stress. We generated skin fibroblasts from the proband and proband's mother (heterozygous) to compare protein expression and subcellular localization of HSP70 under heat stress conditions, and the effect of vector-mediated overexpression of HIKESHI in the proband's cells under the same heat stress conditions. Western blot analysis revealed absent HIKESHI protein from proband fibroblasts, contrasted with ample expression in parental cells. Under heat stress conditions, while the mother's cells displayed appropriate nuclear localization of HSP70, the proband's cells displayed impaired nuclear translocalization. When patient fibroblasts were provided exogenous HIKESHI, the transfected proband's cells showed restored heat-induced nuclear translocalization of HSP70 under conditions of heat stress. These functional data establish that the patient's variant is a pathogenic loss-of-function mutation, thus confirming a diagnosis of hypomyelinating leukodystrophy 13 and that vector-mediated gene replacement may be an effective treatment approach for patients with this disorder.

Microbacterium aquilitoris sp. nov. and Microbacterium gwkjiense sp. nov., isolated from beach.

Three Gram-reaction-positive bacterial strains, designated KSW-18T, KSW2-22, and KSW4-11T, were isolated from seawater, and two dried seaweed samples collected at Gwakji Beach in Jeju, Republic of Korea, respectively, and their taxonomic positions were examined by a polyphasic approach. The 16S rRNA gene phylogeny showed that strain KSW4-11T was tightly associated with Microbacterium oleivorans NBRC 103075T, while strains KSW-18T and KSW2-22 formed a distinctive subline at the base of a clade including the above two strains. The three isolates showed high sequence similarity with one another (99.7-99.9%; 1-4 nt differences) and Microbacterium oleivorans (99.8-99.9%; 1-3 nt differences). The chemotaxonomic features were typical for the genus Microbacterium; Lysine as the diagnostic diamino acid and N-glycolylated muramic acid of the peptidoglycans, the predominant menaquinones of MK-11, MK-10 and MK-12, the major fatty acids of anteiso-C15:0 and anteiso-C17:0, and the major polar lipids including diphosphatidylglycerol, phosphatidylglycerol, and two or three unidentified glycolipids. In core genome-based phylogenetic tree, strains KSW-18T and KSW2-22 were closely associated with Microbacterium oleivorans NBRC 103075T, while strain KSW4-11T formed a distinctive subline at the base of a clade including the above three strains, in contrast to the 16S rRNA gene tree. Strains KSW-18T and KSW2-22 shared an OrthoANIu of 98.6% and a digital DNA-DNA hybridization of 87.6% with each other, representing that they were strains of a species, while the OrthoANIu and digital DNA-DNA hybridization values between strains KSW-18T and KSW4-11T, and between both of these isolates and all members of the genus Microbacterium were ≤86.5% and ≤30.7%, respectively. The analyses of overall genomic relatedness indices and phenotypic distinctness support that the three isolates represent two new species of the genus Microbacterium. Based on the results obtained here, Microbacterium aquilitoris sp. nov. (type strain KSW-18T = KCTC 49623T = NBRC 115222T) and Microbacterium gwkjiense sp. nov. (type strain KSW4-11T = KACC 23321T = DSM 116380T) are proposed.

Gordonibacter faecis sp. nov., producing urolithin C from ellagic acid, isolated from feces of healthy Korean subjects.

A Gram-stain-positive, anaerobic, motile, and short rod-shaped bacterium, designated KGMB12511T, was isolated from the feces of healthy Koreansubjects. Phylogenetic analysis based on the 16S rRNA gene sequence showed that strain KGMB12511T was closely related to Gordonibacter pamelaeae 7-10-1-bT (95.2%). The draft genome of KGMB12511T comprised 33 contigs and 2,744 protein-coding genes. The DNA G + C content was 59.9% based on whole-genome sequences. The major cellular fatty acids (>10%) of strain KGMB12511T were C18:1 cis9, C18:1 cis9 DMA (dimethylacetal), and C16:0 DMA. The predominant polar lipids included a diphosphatydilglycerol, four glycolipids, and an unidentified phospholipid. The major respiratory quinones were menaquinone 6 (MK-6) and monomethylmenaquinone 6 (MMK-6). Furthermore, HPLC analysis demonstrated the ability of strain KGMB12511T to convert ellagic acid into urolithin. Based on a comprehensive analysis of phenotypic, chemotaxonomic, and phylogenetic data, strain KGMB12511T represents a novel species in the genus Gordonibacter. The type strain is KGMB12511T (= KCTC 25343T = NBRC 116190T).

Adlercreutzia faecimuris sp. nov., producing propionate and acetate isolated from mouse feces.

A novel strictly anaerobic bacterium, strain JBNU-10 T, was isolated from BALB/c mouse feces. Cells of the strain JBNU-10 T were Gram-stain positive, non-motile and rod-shaped. Optimum growth occurred at 37℃, with 1% (w/v) NaCl and at pH 7. Phylogenetic analysis based on 16S rRNA gene sequence showed that strain JBNU-10 T belonged to the genus Adlercreutzia and were closely related to Adlercreutzia muris WCA-131-CoC-2 T (95.90%). The genome sequencing of strain JBNU-10 T revealed a genome size of 2,790,983 bp, a DNA G + C content of 69.4 mol%. It contains a total of 2,266 CDSs, 5 rRNA genes and 49 tRNA genes. According to the data obtained strain JBNU-10 T shared ANI value below 77.6- 67.7%, dDDH value below 23.8% with the closely type species. Strain JBNU-10 T possessed iso-C16:0 DMA, C18:1 CIS 9 FAME, and C18:0 DMA as the major fatty acids and had DMMK-6. The major end products of fermentation is propionate and acetate. Based on phylogenetic, physiological and chemotaxonomic characteristics, strain JBNU-10 T represent a novel species of the genus Adlercreutzia. The type strain is JBNU-10 T (= KCTC 25028 T = CCUG 75610 T).

Jatrophihabitans cynanchi sp. nov., isolated from rhizosphere soil of Cynanchum wilfordii.

A novel actinobacterial strain, SB3-54T was isolated from rhizosphere soil of Cynanchum wilfodill, Jaecheon, Chungcheongbuk-do, Republic of Korea. Cells of strain SB3-54T were Gram-stain-positive, aerobic, rod-shaped, and flagellated which formed pale yellow colonies on Reasoner's 2A (R2A) agar. Growth occurred at 15-30 °C (optimum 25 °C), pH 5-8 (optimum pH 7), and 0-2.5% NaCl (optimum 0%). Phylogenetic and phylogenomic analyses showed that strain SB3-54T formed a separate lineage in the genus Jatrophihabitans with Jatrophihabitans telluris N237T. Strain SB3-54T was positive for catalase activity. Genomic analysis showed that SB3-54T has plant-beneficial function contributing (referred to as PBFC) genes such as root colonization and plant protection from oxidative stress. Furthermore, genome of SB3-54T contained gene clusters related to cytokinin biosynthesis, auxin response, tryptophan biosynthesis, siderophore biosynthesis and bacterial toxin-antitoxin systems. Strain SB3-54T contained iso-C16:0 as the major fatty acid and MK-9(H4) and MK-9(H6) as the predominant quinones. The organism had meso-diaminopimelic acid as the diagnostic diamino acid in the peptidoglycan. The major polar lipids were diphosphatidylglycerol, phosphatidylinositol polymannosides, two unidentified aminoglycophospholipids and three unidentified phospholipids. Based on phylogenetic, physiological and chemotaxonomic characteristics, strain SB3-54T represents a novel species of the genus Jatrophihabitans. The type strain is SB3-54T (= KCTC 49134T = NBRC 114108T).

Phocaeicola acetigenes sp. nov., producing acetic acid and iso-butyric acid, isolated faeces from a healthy human.

An obligately anaerobic, non-motile, Gram-stain-negative, and rod-shaped strain KGMB11183T was isolated from the feces of healthy Koreans. The growth of strain KGMB11183T occurred at 30-45 °C (optimum 37 °C), at pH 6-9 (optimum pH 7), and in the presence of 0-0.5% NaCl (optimum 0%). Strain KGMB11183T showed 16S rRNA gene sequence similarities of 95.4% and 94.2% to the closest recognized species, Phocaeicola plebeius M12T, and Phocaeicola faecicola AGMB03916T. Phylogenetic analysis showed that strain KGMB11183T is a member of the genus Phocaeiocla. The major end products of fermentation are acetic acid and isobutyric acid. The major cellular fatty acids (> 10%) of this isolate were C18:1 cis 9, anteiso-C15:0, and summed feature 11 (iso-C17:0 3-OH and/or C18:2 DMA). The assembled draft genome sequences of strain KGMB11183T consisted of 3,215,271 bp with a DNA G + C content of 41.4%. According to genomic analysis, strain KGMB11183T has a number of genes that produce acetic acid. The genome of strain KGMB11183T encoded the starch utilization system (Sus) operon, SusCDEF suggesting that strain uses many complex polysaccharides that cannot be digested by humans. Based on the physiological, chemotaxonomic, phenotypic, and phylogenetic data, strain KGMB11183T is regarded a novel species of the genus Phocaeicola. The type strain is KGMB11183T (= KCTC 25284T = JCM 35696T).

Collinsella urealyticum sp. nov., a urease-positive bacterial strain isolated from swine faeces.

A novel actinobacterial strain, designated AGMB00827T, was isolated from swine faeces. Strain AGMB00827T was obligately anaerobic, Gram-stain-positive, non-motile, non-spore-forming and rod-shaped bacterium. Comparative analyses based on the 16S rRNA gene and whole genome sequence revealed that strain AGMB00827T was affiliated to the genus Collinsella, and was most closely related to Collinsella vaginalis Marseille-P2666T (= KCTC 25056T). Biochemical analysis showed strain AGMB00827T was negative for catalase and oxidase. Interestingly, strain AGMB00827T possessed urease activity, which was determined by traditional methods (API test and Christensen's urea medium), unlike related strains. Furthermore, the major cellular fatty acids (> 10%) of the isolate were C18:1 ω9c, C16:0, C16:0 DMA and C18:2 ω9,12c DMA. Based on the whole genome sequence analysis, the DNA G + C content of strain AGMB00827T was 52.3%, and the genome size and numbers of rRNA and tRNA genes were 1,945,251 bp, 3 and 46, respectively. The average nucleotide identity and digital DNA-DNA hybridization values between strain AGMB00827T and C. vaginalis KCTC 25056 T were 71.0 and 23.2%, respectively. Additionally, the genome analysis revealed that strain AGMB00827T possesses urease gene cluster including ureABC and ureDEFG while the related strains do not have those genes, which is consistent with the urease activity. On the basis of polyphasic taxonomic approach, strain AGMB00827T represents a novel species within the genus Collinsella, for which the name Collinsella urealyticum sp. nov. is proposed. The type strain is AGMB00827T (= KCTC 25287T = GDMCC 1.2724T).

Leeia speluncae sp. nov., isolated from Stegodon Sea Cave in Thailand.

A novel Gram-stain-negative, obligately aerobic, short rod-shaped and motile bacterium, designated strain BC00092T, was isolated from brackish ground water collected in Stegodon Sea Cave located at Satun UNESCO Global Geopark, Satun Province, Thailand. The phylogenetic analysis of BC00092T based on 16S rRNA gene sequences revealed that the strain represented a member of the genus Leeia and was closely related to Leeia oryzae DSM 17879T (96.68 %) and Leeia aquatica IMCC25680T (94.89 %). The average nucleotide identity and digital DNA-DNA hybridization values calculated from the whole-genome sequences between BC00092T and closely related type strains of species within the family Leeiaceae were lower than the species demarcation threshold values of 95 and 70 %, respectively. Moreover, five conserved signature indels of members of the family Leeiaceae were found in the protein sequences from the annotated assembled genome of BC00092T. According to the results of the polyphasic taxonomic study, strain BC00092T represents a novel species within the genus Leeia, for which the name Leeia speluncae sp. nov. is proposed. The type strain is BC00092T (TBRC 13508T = KCTC 92111T).

Actinomadura terrae sp. nov. and Actinomadura litoris OS3-89, isolated from rhizosphere soil of cactus.

Two mycelium-forming actinobacterial strains, designated OS3-83 T and OS3-89, were isolated from rhizosphere soil of a cactus (Opuntia ficus-indica) sampled on Mara Island, Jeju, Republic of Korea. Both of the isolates were found to grow at 20-37 °C, pH 6.0-10.0 and with 0-2 % (w/v) NaCl. Their taxonomic positions were investigated by a polyphasic approach. Strains OS3-83T and OS3-89 were most closely related to the type strain of Actinomadura litoris (99.5 % and 98.9 % 16S rRNA gene sequence similarity, respectively). Both of the isolates shared 99.2 % sequence similarity to each other. Morphological and chemotaxonomic characteristics supported the affiliation of the two isolates to the genus Actinomadura. 16S rRNA gene phylogeny exhibited that strain OS3-83T formed a tight cluster with A. litoris, while strain OS3-89 occupied a position located remotely from A. litoris. Nevertheless, phylogenomic analysis based on 92 core gene sequences showed that both of the isolates formed a tight clade with A. litoris. The values of average nucleotide identity and digital DNA-DNA hybridization between strain OS3-83T and the closest relative, A. litoris, were 92.2 and 46.2 %, respectively, whereas strain OS3-89 shared an average nucleotide identity value of 97.5 % and a digital DNA-DNA hybridization value of 76.9 % with A. litoris. These results strongly suggested that strain OS3-83T (=KACC 19752T=NBRC 114688T) represents a novel species and strain OS3-89 (=KACC 19753=NBRC 114400) is a strain of A. litoris. On the basis of the data obtained here, strain OS3-83T is considered to represent a new species of the genus Actinomadura, for which the name Actinomadura terrae sp. nov. is proposed.

Pseudomonas aestuarii sp. nov., isolated from tidal flat sediment.

Two novel Pseudomonas strains, SA3-5T and SA3-6, were isolated from a tidal flat (getbol) in the Republic of Korea. Strains SA3-5T and SA3-6 were subjected to polyphasic characterization to determine their taxonomic affiliations. Cells were Gram-stain-negative, aerobic, rod-shaped and motile by using peritrichous flagella. Based on their 16S rRNA gene sequences, strains SA3-5T and SA3-6 exhibited a high degree of similarity (100 %) and were classified within the genus Pseudomonas. Furthermore, the closest related species to SA3-5T and SA3-6 were Pseudomonas taeanensis MS-3T (98.3 %). The ranges of average nucleotide identity and digital DNA-DNA hybridization values between SA3-5T and closely related species were 75.9-89.1% and 21.3-38.7%, respectively, both of which being below the thresholds for delineating novel strains. Strain SA3-5T and SA3-6 contained C16 : 1 ω6с and/or C16 : 1 ω7с (summed feature 3), C16 : 0 and C18 : 1 ω6с and/or C18 : 1 ω7с (summed feature 8) as the major fatty acids. The predominant respiratory quinone was Q-9. The DNA G+C content of strain SA3-5T was 62.5 mol%. Based on their combined phenotypic, chemotaxonomic and phylogenetic characterisitics, strains SA3-5T and SA3-6 represent a novel species of the genus Pseudomonas for which the name Pseudomonas aestuarii sp. nov. is proposed. The type strain is SA3-5T (=KCTC 92395T=JCM 35697T).

Caproiciproducens faecalis sp. nov., Isolated from Cow Faeces.

The bacterial strain AGMB10547T was isolated from cow faeces deposited by the National Institute of Animal Science in Cheonan, Republic of Korea. The strain AGMB10547T possessed the phenotypic, biochemical and chemotaxonomic characteristics of the bacteria of the family Oscillospiraceae. The isolate was obligately anaerobic, non-motile, Gram-positive and rod-shaped bacteria. The growth of strain AGMB10547T occurred within 35-40 °C (optimum at 37 °C), at pH 6-7 (optimum of 7) and in the presence of 0.5-2.0% (w/v) NaCl. Based on 16S rRNA gene sequence similarity, strain AGMB10547T belonged to the genus Caproiciproducens and was most closely related to Caproiciproducens galactitolivorans BS-1T (96.9%). The DNA G+C content was 49.0 mol%. The major cellular fatty acids (> 10%) of the isolate were C14:0, C14:0 DMA, C16:1 ω9c and C16:0. The average nucleotide identity (ANI) and digital DNA-DNA Hybridization (dDDH) values between strain AGMB10547T and C. galactitolivorans BS-1T were 75.5% and 19.2%. Based on the phenotypic, genotypic, biochemical and chemotaxonomic analyses, strain AGMB10547T represents a novel species of the genus Caproiciproducens, for which the name Caproiciproducens faecalis sp. nov. is proposed. The type strain AGMB10547T (=KCTC 25200T=NBRC 115006T=GDMCC 1.2575T).

Parabacteroides faecalis sp. nov. Isolated from Swine Faeces.

A Gram-negative, obligate anaerobic, non-motile, non-spore-forming, rod-shaped bacterial strain designated AGMB00274T was isolated from swine faeces. An 16S rRNA gene analysis indicated that strain AGMB00274T belonged to the genus Parabacteroides, with the highest similarity to Parabacteroides johnsonii (P. johnsonii) DSM 18315T (sequence similarity of 94.9%). The genome size of strain AGMB00274T was 4,308,683 bp, with a DNA G+C content of 42.5 mol%. The biochemical analysis of strain AGMB00274T showed that it was positive for gelatin hydrolysis and α-fucosidase, but negative for the acid production from D-glucose, D-mannitol, D-maltose, salicin, glycerol, D-cellobiose, D-mannose, D-melezitose, D-sorbitol, D-trehalose, and negative for α-arabinosidase, glutamic acid decarboxylase, and pyroglutamic acid arylamidase. The dominant cellular fatty acids (> 10%) of the isolate were anteiso-C15: 0 (23.2%), iso-C15: 0 (16.6%), C18: 1 ω9c (16.4%), summed feature 11 (iso-C17: 0 3-OH and/or C18: 2 DMA) (12.5%), and C16: 0 (11.3%). The major respiratory quinones of strain AGMB00274T were MK-9 (55.4%) and MK-10 (44.6%). The major polar lipid was phosphatidylethanolamine. Based on phylogenetic, genetic, physiological, and chemotaxonomic analyses, as a novel species of the genus Parabacteroides, strain AGMB00274T was proposed with the name Parabacteroides faecalis sp. nov. The type strain used was AGMB00274T (= KCTC 25286T = GDMCC 1.2742T).

Effectiveness of Project ECHO Programs in Improving Clinician Knowledge and Confidence in Managing Complex Psychiatric Patients: a Waitlist-Controlled Study.

OBJECTIVE: The authors aimed to determine if Project Extension for Community Healthcare Outcomes (ECHO), a health-education model utilising teleconferencing technology, improves the capacity of clinicians in assessing and managing complex psychiatric patients. METHODS: Three pilot Project ECHO programs were evaluated as a prospective waitlist-controlled trial, focusing on Adult Eating Disorders, Adult Intellectual Disability Mental Health, and General Mental Health. Each program comprised 9-10 weekly teleconferencing group sessions. Participants and waitlist-controls completed pre- and post-program surveys. The primary outcomes were self-reported knowledge and confidence in assessing and managing complex patients relevant to each group. Linear mixed models were used to assess the group-by-time interaction, or change over time, as appropriate. RESULTS: Between July 2020 and June 2021, three series of the Adult Intellectual Disability Mental Health program, two series of the Adult Eating Disorders program, and two series of the General Mental Health program were delivered. Compared to waitlist-controls (n = 21), there were statistically significant improvements in self-reported knowledge and confidence for all topics amongst participants of the Adult Eating Disorders program (n = 44). In the Adult Intellectual Disability Mental Health program, there were significant improvements in self-reported knowledge and confidence amongst participants (n = 67) for most topics compared to controls (n = 21). There were no waitlist-controls for the General Mental Health program, but within-group analysis (n = 28) showed significant improvements in participants' knowledge and confidence following program completion, compared to baseline. CONCLUSION: Project ECHO is a feasible and effective model to develop workforce capacity in managing complex psychiatric conditions.

TBX6 as a cause of a combined skeletal-kidney dysplasia syndrome.

TBX6 encodes transcription-factor box 6, a transcription factor critical to paraxial mesoderm segmentation and somitogenesis during embryonic development. TBX6 haploinsufficiency is believed to drive the skeletal and kidney phenotypes associated with the 16p11.2 deletion syndrome. Heterozygous and biallelic variants in TBX6 are associated with vertebral and rib malformations (TBX6-associated congenital scoliosis) and spondylocostal dysostosis, and heterozygous TBX6 variants are associated with increased risk of genitourinary tract malformations. Combined skeletal and kidney phenotypes in individuals harboring heterozygous or biallelic TBX6 variants are rare. Here, we present seven individuals with vertebral and rib malformations and structural kidney differences associated with heterozygous TBX6 gene deletion in trans with a hypomorphic TBX6 allele or biallelic TBX6 variants. Our case series highlights the association between TBX6 and both skeletal and kidney disease.

Bacteroides humanifaecis sp. nov., isolated from faeces of healthy Korean.

Two obligately anaerobic, Gram-stain-negative, non-motile, non-spore-forming and short rod shaped bacteria, designated KGMB07931T and KGMB10229, were isolated from faeces of two Korean persons. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strains KGMB07931T and KGMB10229 were very similar to each other (99.9%) and grouped within the genus Bacteroides, displaying the highest similarity with Bacteroides uniformis ATCC 8492T (97.5%), Bacteroides rodentium JCM 16496T (96.6%), and Bacteroides fluxus YIT 12057T (94.5%). Both strains grew optimally at 37 °C and pH 7.5 in the presence of 0.5% (w/v) NaCl. The complete genome of KGMB07931T comprises 3,335 protein-coding genes with a total of 4,240,638 bp and an average G + C content of 46.3 mol%. The major fatty acids were C18:1 cis9, anteiso-C15:0, iso-C15:0, and Summed Feature 11 (iso-C17:0 3OH and/or C18:2 DMA); the predominant respiratory quinones were MK-9 and MK-10; the major fermentation end products were acetate and isobutyrate. The genome of strain KGMB07931T encoded the starch utilization system (Sus) operon, susABCDEFG, suggesting that this strain uses many complex polysaccharides that cannot be digested by humans. Based on polyphasic taxonomic data, strains KGMB07931T and KGMB10229 represent a novel species within the genus Bacteroides, for which the name Bacteroides humanifaecis sp. nov. is proposed. The type strain is KGMB07931T (= KCTC 25160T = NBRC 115005T).

Paenibacillus vietnamensis sp. nov., isolated from the rhizosphere soil of Arachis hypogaea.

A novel bacterial strain, N4T, was isolated from the soil of a groundnut Arachis hypogaea field in Nghean province, Vietnam. The phylogenetic, chemotaxonomic and phenotypic characteristics of this strain were determined. Cells of strain N4T were Gram-negative, aerobic, endospore-forming and rod-shaped. Strain N4T grew at 20-37 °C (optimum, 30 °C), pH 6-10 (optimum, pH 7) and 0-5 % NaCl (optimum, 0 %). Phylogenetic analysis based on 16S rRNA gene sequences showed that strain N4T belonged to the genus Paenibacillus and was closely related to Paenibacillus harenae B519T (=KCTC 3951T) and Paenibacillus alkaliterrae KSL-134T (=KCTC 3956T) with 96.3 and 96.5% gene sequence similarity, respectively. It contained meso-diaminopimelic acid as the diagnostic diamino acid in the cell-wall peptidoglycan. The DNA G+C content was 52.9 mol%. The major isoprenoid quinone was MK-7. Anteiso-C15 : 0 and iso-C16 : 0 were the dominant cellular fatty acids. Based on phylogenetic, physiological and biochemical characteristics, strain N4T represents a novel species of the genus Paenibacillus, for which the name Paenibacillus vietnamensis sp. nov. is proposed. The type strain is N4T (=KCTC 33932T= VTCC 12236T).

Succinivibrio faecicola sp. nov., isolated from cow faeces.

A Gram-stain-negative, obligately anaerobic, non-motile, non-spore-forming, helical rod-shaped bacterium, designated AGMB01872T, was isolated from faeces of a cow deposited in the National Institute of Animal Science (Wanju, Republic of Korea). Phylogenetic analysis based on the 16S rRNA gene sequences showed that strain AGMB01872T was most closely related to Succinivibrio dextrinosolvens DSM 3072T (= KCTC 25222T, 96.6 %) which belonged to the family Succinivibrionaceae. Growth was occurred at 30-40 °C (optimum, 37 °C), pH 6-7 (optimum, pH 7) and in the presence of 0.5-1.0 % (w/v) NaCl. The genomic DNA G+C content of strain AGMB01872T was 35.9 mol%. The average nucleotide identity value between strain AGMB01872T and S. dextrinosolvens DSM 3072T was 72.1 %. Cells of strain AGMB01872T utilized d-glucose, maltose, d-xylose and l-arabinose. The major fatty acids (>10 %) were C14 : 0 (23.9 %), C16 : 0 (29.4 %), summed feature 5 (10.8 %) and summed feature 10 (30.3 %). The major end-product of glucose fermentation was succinate. Based on the phenotypic, phylogenetic, biochemical, genotypic and chemotaxonomic data, AGMB01872T represents a novel species within the genus Succinivibrio, for which the name Succinivibrio feacicola sp. nov. is proposed. The type strain is AGMB01872T (= KCTC 25201T=NBRC 115007T=GDMCC 1.2573T).

Nicotine Rather Than Non-Nicotine Substances in 3R4F WCSC Increases Behavioral Sensitization and Drug-Taking Behavior in Rats.

INTRODUCTION: Nicotine increases reinforcing effects of cigarette smoking by upregulating glutamate and dopamine releases via stimulation of nicotinic acetylcholine receptors (nAChRs) in the dorsal striatum (CPu). The present study was conducted to evaluate whether non-nicotine substances in cigarette smoke potentiate nicotine-induced behaviors by increasing glutamate and dopamine concentrations in the CPu. AIMS AND METHODS: Changes in the levels of glutamate and dopamine in the CPu were analyzed using a glutamate colorimetric assay and dopamine enzyme-linked immunosorbent assay, respectively, after repeated administration of nicotine or whole cigarette smoke condensate (WCSC) in male Sprague-Dawley rats. Changes in locomotion and drug-taking behavior were analyzed using the measurements of locomotor activity and self-administration under a fixed ratio 1 schedule in response to repeated administration of nicotine or WCSC. RESULTS: Repeated subcutaneous (s.c.) injections of nicotine (0.25 mg/kg/day) for 7 consecutive days significantly increased the levels of glutamate and dopamine in the CPu. Similar results were obtained from repeated injections of WCSC (0.25 mg/kg nicotine/day, s.c.) extracted from 3R4F Kentucky reference cigarettes. Parallel with the increases in the neurotransmitter levels in the CPu, both nicotine and WCSC increased locomotor activity and self-administration (0.03 mg/kg nicotine/infusion). However, repeated injections of WCSC did not change the nicotine-induced increases in neurotransmitter levels, locomotor activity, and self-administration. CONCLUSIONS: Nicotine rather than non-nicotine substances in WCSC play a major role in potentiating behavioral sensitization and drug-taking behavior via elevation of glutamate and dopamine concentrations in the CPu of rats. IMPLICATIONS: WCSC does not augment the nicotine-induced increases in behavioral sensitization, drug-taking behavior, and glutamate and dopamine concentrations, suggesting that non-nicotine substances do not potentiate the nicotine-induced behaviors by increasing the concentrations of the neurotransmitters in the CPu. These findings imply that nicotine, but not non-nicotine substances in WCSC, may be a major contributor that induces tobacco dependence in rats.

Oncologic Outcomes of Nipple-sparing Mastectomy and Immediate Reconstruction After Neoadjuvant Chemotherapy for Breast Cancer.

OBJECTIVES: To evaluate the oncologic outcomes and risk factors for locoregional recurrence (LRR) and nipple-areola complex recurrence (NR) in a large series of breast cancer patients who underwent nipple-sparing mastectomy (NSM) and immediate reconstruction after neoadjuvant chemotherapy (NACT). SUMMARY OF BACKGROUND DATA: The use of NSM and immediate reconstruction in breast cancer patients receiving NACT is increasing. However, the oncologic safety of this approach is unclear. PATIENTS AND METHODS: A total of 310 breast cancer patients (319 breasts) who underwent NACT and NSM between February 2010 and November 2016 were retrospectively analyzed. Clinical and pathologic factors associated with increased risks of LRR and NR were analyzed using univariate (Chi-square or Fisher exact test) and multivariate (Cox proportional hazard regression model) analyses. RESULTS: During a mean follow-up of 63 ± 22 months, 38 cases had LRR as the first event, including 6 cases of NR as the first event. The 5-year cumulative LRR and NR rates were 11.0% and 1.9%, respectively. In univariate analysis, clinical T stage, pathologic nodal status, histologic grade, lymphovascular invasion, and post-NACT Ki67 status were associated with increased LRR risk, and post-NACT Ki67 status was the only significant risk factor for NR. In multivariate analysis, post-NACT Ki67 ≥10% (hazard ratio, 4.245; 95% confidence interval, 1.865-9.663; P = 0.001) was an independent risk factor for LRR. CONCLUSIONS: NSM and immediate reconstruction seem to be oncologically safe with acceptable LRR and NR rates for appropriately selected breast cancer patients treated with NACT. Post-NACT Ki67 ≥10% was associated with increased risk of LRR or NR, and therefore, necessitates cautious follow-up.

Engineering of an EpCAM-targeting cyclic peptide to improve the EpCAM-mediated cellular internalization and tumor accumulation of a peptide-fused antibody.

Fusion of a target-specific peptide to a full-length antibody (Ab) can result in a peptide-Ab fusion protein with additional specificity and enhanced activity. We recently developed an intracellular pan-RAS-targeting cytosol-penetrating antibody, RT22-ep59, in which a tumor-specific targeting ability was achieved via the fusion of an epithelial cell adhesion molecule (EpCAM) targeting cyclic peptide (ep133). Here, the aim was to enhance EpCAM-mediated endocytosis and tumor accumulation of the peptide-fused RAS-targeting Ab. Accordingly, we engineered a cyclic peptide (from ep133) that has stronger affinity for EpCAM by using yeast surface display technology and then rationally designed cyclic peptides in the Ab-fused form to enhance colloidal stability. The finally engineered EpCAM-targeting cyclic peptide (ep6)-fused Ab, ep6Ras37, has ∼10-fold stronger affinity (KD ≈ 1.9 nM) for EpCAM than that of RT22-ep59, without deterioration of biophysical properties. Compared with the parental antibody (RT22-ep59), ep6Ras37 more efficiently reached the cytosol of EpCAM-expressing cells and showed greater preferential tumor homing and accumulation in mice bearing EpCAM-expressing LoVo xenograft tumors. Thus, the high-affinity EpCAM-targeting peptide ensures efficient cellular internalization and better tumor accumulation of the peptide-fused Ab.