Development of a simple and practical method of discrimination between Vibrio furnissii and V. fluvialis based on single-nucleotide polymorphisms of 16S rRNA genes observed in V. furnissii but not in V. fluvialis.

Vibrio furnissii and V. fluvialis are closely related, the discrimination of which by conventional biochemical assay remains a challenge. Investigation of the sequence of the 16S rRNA genes in a clinical isolate of V. furnissii by visual inspection of a sequencing electropherogram revealed two sites of single-nucleotide polymorphisms (SNPs; positions 460 A/G and 1261 A/G) in these genes. A test of 12 strains each of V. fluvialis and V. furnissii revealed these SNPs to be common in V. furnissii but not in V. fluvialis. Divergence of SNP frequency was observed among the strains of V. furnissii tested. Because the SNPs described in V. furnissii produce a difference in the target sequence of restriction enzymes, a combination of polymerase chain reaction (PCR) of the 16S rRNA genes using conventional primers and restriction fragment length polymorphism analysis using Eco RV and Eae I was shown to discriminate between V. fluvialis and V. furnissii. This method is simple and alleviates the need for expensive equipment or primer sets specific to these bacteria. Therefore, we believe that this method can be useful, alongside specific PCR and mass spectrometry, when there is a need to discriminate between V. fluvialis and V. furnissii.

Angiomyofibroblastoma of the vulva: report of a case with immunohistochemical and molecular analysis.

A 43-year-old woman presented with a mass in the subcutaneous tissue of the right labium majus. A lipoma or Bartholin gland cyst was suspected and excision of the lesion was performed. The lesion was well circumscribed, and histological examination revealed a typical angiomyofibroblastoma. The lesion was composed of alternating hypocellular edematous and hypercellular areas with abundant vessels, and plump tumor cells were loosely dispersed or aggregated mainly around the vessels. Tumor cells were immunoreactive for vimentin and desmin but negative for muscle actins. Ultrastructurally, the tumor cells contained a moderate amount of rough endoplasmic reticulum and abundant intermediate filaments, and had primitive junctions. Pinocytotic vesicles or basal lamina were not evident. Immunohistochemical studies also revealed that the tumor cells expressed basic fibroblast-growth factor, vascular-endothelial-growth factor, and stem-cell factor, factors that may contribute to the rich vascularity and mast cells within the tumor. Reverse transcription-polymerase chain reaction detected high mobility group I-C (HMGI-C) transcripts in the tumor tissue. Because the expression of HMGI-C is regulated by developmental and differentiation processes and is not found in adult normal tissues, HMGI-C may be involved in the tumorigenesis of angiomyofibroblastoma.