A phage protein-derived antipathogenic peptide that targets type IV pilus assembly.

Phage-inspired antibacterial discovery is a new approach that recruits phages in search for antibacterials with new molecular targets, in that phages are the biological entities well adapted to hijack host bacterial physiology in favor of their own thrive. We previously observed that phage-mediated twitching motility inhibition was effective to control the acute infections caused by Pseudomonas aeruginosa and that the motility inhibition was attributed to the delocalization of PilB, the type IV pilus (TFP) assembly ATPase by binding of the 136-amino acid (aa) phage protein, Tip. Here, we created a series of truncated and point-mutant Tip proteins to identify the critical residues in the Tip bioactivity: N-terminal 80-aa residues were dispensable for the Tip activity; we identified that Asp82, Leu84, and Arg85 are crucial in the Tip function. Furthermore, a synthetic 15-aa peptide (P1) that corresponds to Leu73 to Ala87 is shown to suffice for PilB delocalization, twitching inhibition, and virulence attenuation upon exogenous administration. The transgenic flies expressing the 15-aa peptide were resistant to P. aeruginosa infections as well. Taken together, this proof-of-concept study reveals a new antipathogenic peptide hit targeting bacterial motility and provides an insight into antibacterial discovery targeting TFP assembly.

Pseudocnuella soli gen. nov., sp. nov., a bacterium from soil belonging to the family Chitinophagaceae.

A Gram-stain negative, aerobic, rod-shaped, yellow-coloured bacterium, designated strain 17J28-1T, was isolated from soil of Jeju Island, Korea. Optimal growth was found to occur at 30 °C, at pH 6.0 and in the absence of NaCl on R2A at 30 °C. Strain 17J28-1T showed resistance to UV-radiation. The draft genome sequence of strain 17J28-1T is 4804,510 bp long with a 49.4% G + C content. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain 17J28-1T forms a distinct lineage within the family Chitinophagaceae (order Chitinophagales, class Chitinophagia) and is closely related to Cnuella takakiae (93.7% 16S rRNA gene sequence similarity) and Flavisolibacter ginsengiterrae (93.1%). In addition, genomic comparison using digital DNA-DNA hybridization (dDDH) and OrthoANI analyses between the novel organism and the members of the family Chitinophagaceae revealed identities of 65.5-74.1% and 18.1-28%, respectively. The predominant cellular fatty acids were identified as C15:0 iso, C15:1 iso G and C17:0 iso 3-OH and the major respiratory quinone as MK-7. Based on these data we propose that strain 17J28-1T (= KCTC 62223T = JCM 33202T) represents a novel genus and species in the family Chitinophagaceae, for which the name Pseudocnuella soli gen. nov. sp. nov. is proposed.

Sphingomonas parva sp. nov., isolated from soil in Jeju Island.

A Gram staining-negative, yellow-colored, rod-shaped, non-motile and aerobic, designated strain 17J27-24T was isolated from a soil sample in Korea. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain 17J27-24T was related to the members of the family Sphingomonadaceae and formed a distinct monophyletic cluster within the genus Sphingomonas with Sphingomonas deserti (98.3% 16S rRNA gene sequence similarity). Growth was observed at 30 °C (optimum), at pH 7.0 (optimum), and in the absence of NaCl (%). The predominant cellular fatty acids were summed feature 8 (18:1 ω7c and/or 18:1 ω6c) and C17:1 ω6c. The major respiratory quinone was Q10. The polar lipids profile comprised of diphosphatidylglycerol (DPG), phosphatidylglycerol (PG), phosphatidylethanolamine (PE), and sphingoglycolipid (SGL). The DNA G + C content was 77.8 mol%. The average nucleotide identity (ANI) and digital DNA-DNA hybridisation (dDDH) values between 17J27-24T and its phylogenetically closest Sphingomonas deserti (KCTC 62411T) were below the established cut-off < 94% (ANI) and < 70% (dDDH) for species delineation. Moreover, the results of the polyphasic approach confirmed that strain 17J27-24T represents a novel species of the genus Sphingomonas within the family Sphingomonadaceae, for which the name Sphingomonas parva sp. nov. is proposed. The type strain of this species is 17J27-24T (= KCTC 62208T = JCM 3896T). An emended description of the species Sphingomonas parva is provided.

Occult lymph node metastasis and risk of regional recurrence in papillary thyroid cancer after bilateral prophylactic central neck dissection: A multi-institutional study.

BACKGROUND: The impact of occult lymph node metastasis on regional recurrence after prophylactic central neck dissection for preoperative, nodal-negative papillary thyroid cancer is controversial. We investigated risk factors for regional lymph node recurrence in papillary thyroid cancer patients who underwent total thyroidectomy and bilateral prophylactic central neck dissection. Analysis was according to clinicopathologic characteristics and occult lymph node metastasis patterns. METHODS: This multicenter study enrolled 211 consecutive patients who underwent total thyroidectomy with bilateral prophylactic central neck dissection for papillary thyroid cancer without evidence of central lymph node metastasis on preoperative imaging. Clinicopathologic features and central lymph node metastasis patterns were analyzed for predicting regional recurrence. Multivariate Cox regression analysis was used to identify independent factors for recurrence. RESULTS: Median follow-up time was 43 months (24-95 months). Ten patients (4.7%) showed regional lymph node recurrence. The estimated 5-year, regional recurrence-free survival was 95.2%. Tumor size ≥1 cm, central lymph node metastasis, lymph node ratio, and prelaryngeal lymph node metastasis were associated with regional recurrence in univariate analysis (P < .05). In multivariate analysis, a lymph node ratio ≥ 0.26 was a significant risk factor for regional lymph node recurrence (odds ratio = 11.63, P = .003). Lymph node ratio ≥ 0.26 was an independent predictor of worse recurrence-free survival on Cox regression analysis (hazard ratio = 11.49, P = .002). CONCLUSION: Although no significant association was observed between the presence of occult lymph node metastasis and regional recurrence, lymph node ratio ≥ 0.26 was an independent predictor of regional lymph node recurrence in papillary thyroid cancer patients who underwent total thyroidectomy and bilateral prophylactic central neck dissection.