Rapid direct identification of Cryptococcus neoformans from pigeon droppings by nested PCR using CNLAC1 gene.

Isolation and identification of Cryptococcus neoformans and pathogenic yeast-like fungi from pigeon droppings has been taken for a long time and requires various nutrients for its growth. In this study, we attempted to establish a rapid direct identification method of Cr. neoformans from pigeon dropping samples by nested-PCR using internal transcribed spacer (ITS) CAP64 and CNLAC1 genes, polysaccharide capsule gene and laccase-associated gene to produce melanin pigment, respectively, which are common genes of yeasts. The ITS and CAP64 genes were amplified in all pathogenic yeasts, but CNLAC1 was amplified only in Cr. neoformans. The ITS gene was useful for yeast genotyping depending on nucleotide sequence. Homology of CAP64 genes among the yeasts were very high. The specificity of PCR using CNLAC1 was demonstrated in Cr. neoformans environmental strains but not in other yeast-like fungi. The CNLAC1 gene was detected in 5 serotypes of Cr. neoformans. The nested-PCR amplified up to 10(-11) µg of the genomic DNA and showed high sensitivity. All pigeon droppings among 31 Cr. neoformans-positive samples were positive and all pigeon droppings among 348 Cr. neoformans-negative samples were negative by the direct nested-PCR. In addition, after primary enrichment of pigeon droppings in Sabouraud dextrose broth, all Cr. neoformans-negative samples were negative by the nested-PCR, which showed high specificity. The nested-PCR showed high sensitivity without culture of pigeon droppings. Nested-PCR using CNLAC1 provides a rapid and reliable molecular diagnostic method to overcome weak points such as long culture time of many conventional methods.

Onset of labor epidural analgesia with ropivacaine and a varying dose of fentanyl: a randomized controlled trial.

BACKGROUND: This study was conducted to investigate the onset of labor epidural analgesia using 0.17% ropivacaine with a varying dose of fentanyl. We hypothesized that the onset of analgesia would be shortened in proportion to an increase in fentanyl dose. METHODS: Women requesting labor epidural analgesia were enrolled in this randomized controlled clinical trial. Each woman was randomly assigned to receive fentanyl 0, 50, 75, or 100 µg with 0.17% ropivacaine 12 mL. The onset and duration of analgesia, the incidence of side effects and patient satisfaction were measured. RESULTS: Data from 102 women were analyzed. The onset of analgesia (mean ± SD) was shortened with an increasing dose of fentanyl (14.3 ± 5.4, 14.2 ± 6.5, 12.1 ± 5.1, and 8.7 ± 3.8 min with fentanyl 0, 50, 75, or 100 µg, respectively, P=0.001). The duration of analgesia was prolonged with an increasing dose of fentanyl (87.4 ± 20.8, 112.3 ± 19.5, 140.8 ± 18.8, and 143.6 ± 18.6 min with fentanyl 0, 50, 75, or 100 µg, respectively, P<0.001). The incidence of pruritus increased with an increasing dose of fentanyl (P=0.027) but there were no differences for other maternal side effects. There was a significant difference in satisfaction score among groups (P=0.009). CONCLUSION: The addition of increasing doses of fentanyl to 0.17% ropivacaine contributed to shortened onset as well as prolonged duration of labor epidural analgesia and improved patient satisfaction.

A case of retroperitoneal lipoleiomyoma.

We report a case of lipoleiomyoma which arose in retroperitoneum and presented with progressively distended abdomen in a 56-yr-old woman. The tumor was well encapsulated and consisted of two components, benign smooth muscle cells and mature adipose tissue without any atypia. It is likely to be mistaken as extrarenal angiomyolipoma, well-differentiated liposarcoma and leiomyoma with fatty change. We review the histologic characteristics of previously reported myolipoma and describe essential points of differential diagnosis.

Anticancer activity of hydrophobic peptides from soy proteins.

An anticancer peptide from soy protein was purified and isolated. Defatted soy protein was hydrolyzed with thermoase and hydrophobic peptides were extracted with ethanol. The peptide extract was fractionated by XAD-2 hydrophobic, gel filtration chromatography, and different C18 HPLCs. Anticancer activity of each fraction was assayed by measuring in vitro cytotoxicity on P388D1, a mouse monocyte macrophage cell line. IC50 value of a peptide fraction from Sephadex G-25 chromatography was 0.16 mg/ml. This peptide fraction at 1 mg/ml significantly affected cell cycle progression by arresting P388D1 at G2/M phases. Finally purified peptide from analytical C18 HPLC was nonapeptide of which molecular weight was 1157 Da and the sequence was X-Met-Leu-Pro-Ser-Tye-Ser-Pro-Tyr.