Whole-genome analysis of human papillomavirus genotypes 52 and 58 isolated from Japanese women with cervical intraepithelial neoplasia and invasive cervical cancer

BackgroundHuman papillomavirus genotypes 52 and 58 (HPV52/58) are frequently detected in patients with cervical intraepithelial neoplasia (CIN) and invasive cervical cancer (ICC) in East Asian countries including Japan. As with other HPV genotypes, HPV52/58 consist of multiple lineages of genetic variants harboring less than 10% differences between complete genome sequences of the same HPV genotype. However, site variations of nucleotide and amino acid sequences across the viral whole-genome have not been fully examined for HPV52/58. The aim of this study was to investigate genetic variations of HPV52/58 prevalent among Japanese women by analyzing the viral whole-genome sequences.MethodsThe entire genomic region of HPV52/58 was amplified by long-range PCR with total cellular DNA extracted from cervical exfoliated cells isolated from Japanese patients with CIN or ICC. The amplified DNA was subjected to next generation sequencing to determine the complete viral genome sequences. Phylogenetic analyses were performed with the whole-genome sequences to assign variant lineages/sublineages to the HPV52/58 isolates. The variability in amino acid sequences of viral proteins was assessed by calculating the Shannon entropy scores at individual amino acid positions of HPV proteins.ResultsAmong 52 isolates of HPV52 (CIN1, n = 20; CIN2/3, n = 21; ICC, n = 11), 50 isolates belonged to lineage B (sublineage B2) and two isolates belonged to lineage A (sublineage A1). Among 48 isolates of HPV58 (CIN1, n = 21; CIN2/3, n = 19; ICC, n = 8), 47 isolates belonged to lineage A (sublineages A1/A2/A3) and one isolate belonged to lineage C. Single nucleotide polymorphisms specific for individual variant lineages were determined throughout the viral genome based on multiple sequence alignments of the Japanese HPV52/58 isolates and reference HPV52/58 genomes. Entropy analyses revealed that the E1 protein was relatively variable among the HPV52 isolates, whereas the E7, E4, and L2 proteins showed some variations among the HPV58 isolates.ConclusionsAmong the HPV52/58-positive specimens from Japanese women with CIN/ICC, the variant distributions were strongly biased toward lineage B for HPV52 and lineage A for HPV58 across histological categories. Different patterns of amino acid variations were observed in HPV52 and HPV58 across the viral whole-genome

The relationship between torsion and of the length of the peduncle in patients with uterine subserosal leiomyoma

Objective: To clarify the characteristics of twistable pedunculated subserosal uterine leiomyomas. Methods: A retrospective study was conducted of female subserosal uterine leiomyoma patients who were treated by laparoscopic myomectomy at Showa University Hospital, Tokyo, Japan, between January 2011 and September 2015. The patients were classified according to their intraoperative findings. The maximum diameter and volume of the subserosal uterine leiomyoma, and the length and thickness of the peduncle were evaluated by magnetic resonance imaging, and torsion and nontorsion cases were compared. Results: A total of eight torsion cases and 20 nontorsion cases were analyzed. The average peduncle length was significantly higher in the torsion cases (18.7±3.5 mm) than in the nontorsion cases (9.0±3.9 mm). The maximum diameter and volume of subserosal uterine leiomyoma and the thickness of the peduncle did not differ between the groups. The length to thickness ratio (length to thickness index) was more clearly associated with the torsion than the length of the peduncle or the ratio of the maximum diameter of the leiomyoma to the length. Receiver operating characteristic curve analysis showed that a length to thickness ratio of > 1.0 was associated with a higher detection rate, with an area under the receiver operating characteristic curve of 0.95 (95% confidence interval 0.86–1.00, p<0.001). Conclusions: In the preoperative period, the length of the peduncle was associated with the subsequent occurrence of torsion in patients with subserosal uterine leiomyoma. The length to thickness index is useful for predicting the torsion of a subserosal uterus leiomyoma

Development of Surface Fluorescence X‐Ray Absorption Fine Structure Spectroscopy Using a Laue‐Type Monochromator

Surface fluorescence X-ray absorption fine structure (XAFS) spectroscopy using a Laue-type monochromator has been developed to acquire structural information about metals with a very low concentrate on a flat highly oriented pyrolytic graphite (HOPG) surface in the presence of electrolytes. Generally, surface fluorescence XAFS spectroscopy is hindered by strong scattering from the bulk, which often chokes the pulse counting detector. In this work, we show that a bent crystal Laue analyzer (BCLA) can efficiently remove the scattered X-rays from the bulk even in the presence of solution. We applied the technique to submonolayer (similar to 10(14) atoms cm(-2)) Pt on HOPG and successfully obtained high signal/noise in situ XAFS data in combination with back-illuminated fluorescence XAFS (BI-FXAFS) spectroscopy. This technique allows in situ XAFS measurements of flat electrode surfaces to be performed in the presence of electrolytes

Crystal Structures of S‑Shaped Phenylenediurea Dibenzoic Acids and Their Cocrystals with Melamine: Unusual Zigzag Tape of H‑Bonded Melamine Network

Phenylenediurea dibenzoic acid derivatives <b>1</b>, <b>2</b>, <b>3</b>, and <b>5</b> and an amide derivative <b>4</b> were synthesized, and their molecular structures were elucidated to be an S-shape by single crystal X-ray analysis. Crystals of <b>1</b> were obtained as a DMF solvate, while those of <b>2</b> and <b>3</b> were not. Sheet and ladder structures were created in the crystal packing of <b>1</b> and <b>3</b>, and <b>2</b>, respectively. Unlike the para-substituted diureas <b>1</b>–<b>3</b> which possessed meso-conformation, the meta-substituted diurea <b>5</b> exhibited helical-conformation in its crystal structure. The way of crystal packing of <b>4</b> is analogous to that of the corresponding dicarboxylic acid derivative <b>2</b>. A water cluster of a chair-shaped hexagonal array of water molecules was created in the crystal structure of <b>4</b>. Diureas <b>1</b> and <b>2</b> gave cocrystals with melamine recrystallized from DMF/ethyl acetate and DMF/H₂O, respectively. Unusual zigzag tapes of a H-bonding network of melamine were created in both cocrystals. The zigzag tapes were connected with S-shaped diureas by H-bonding to furnish an interpenetrated fishnet-type crystal structure

Whole-genome analysis of human papillomavirus genotypes 52 and 58 isolated from Japanese women with cervical intraepithelial neoplasia and invasive cervical cancer

Abstract Background Human papillomavirus genotypes 52 and 58 (HPV52/58) are frequently detected in patients with cervical intraepithelial neoplasia (CIN) and invasive cervical cancer (ICC) in East Asian countries including Japan. As with other HPV genotypes, HPV52/58 consist of multiple lineages of genetic variants harboring less than 10% differences between complete genome sequences of the same HPV genotype. However, site variations of nucleotide and amino acid sequences across the viral whole-genome have not been fully examined for HPV52/58. The aim of this study was to investigate genetic variations of HPV52/58 prevalent among Japanese women by analyzing the viral whole-genome sequences. Methods The entire genomic region of HPV52/58 was amplified by long-range PCR with total cellular DNA extracted from cervical exfoliated cells isolated from Japanese patients with CIN or ICC. The amplified DNA was subjected to next generation sequencing to determine the complete viral genome sequences. Phylogenetic analyses were performed with the whole-genome sequences to assign variant lineages/sublineages to the HPV52/58 isolates. The variability in amino acid sequences of viral proteins was assessed by calculating the Shannon entropy scores at individual amino acid positions of HPV proteins. Results Among 52 isolates of HPV52 (CIN1, n = 20; CIN2/3, n = 21; ICC, n = 11), 50 isolates belonged to lineage B (sublineage B2) and two isolates belonged to lineage A (sublineage A1). Among 48 isolates of HPV58 (CIN1, n = 21; CIN2/3, n = 19; ICC, n = 8), 47 isolates belonged to lineage A (sublineages A1/A2/A3) and one isolate belonged to lineage C. Single nucleotide polymorphisms specific for individual variant lineages were determined throughout the viral genome based on multiple sequence alignments of the Japanese HPV52/58 isolates and reference HPV52/58 genomes. Entropy analyses revealed that the E1 protein was relatively variable among the HPV52 isolates, whereas the E7, E4, and L2 proteins showed some variations among the HPV58 isolates. Conclusions Among the HPV52/58-positive specimens from Japanese women with CIN/ICC, the variant distributions were strongly biased toward lineage B for HPV52 and lineage A for HPV58 across histological categories. Different patterns of amino acid variations were observed in HPV52 and HPV58 across the viral whole-genome