Human Papillomavirus Test for Triage of Japanese Women With Low-Grade Squamous Intraepithelial Lesions.

We evaluated high-risk human papillomavirus (HR-HPV) DNA testing for high-grade cervical intraepithelial neoplasia (CIN) lesions by cobas HPV test and diagnostic HPV16/18 genotyping in Japanese women with low-grade squamous intraepithelial lesions. Of 357 patients, HR-HPV positivity prevalence was 75.6%, and 21.3% had grade 2 or higher CIN lesions (CIN2+), with the highest prevalence at 30 to 34 years. Negative predictive values of HR-HPV for CIN2+ in our patients were 93.1% (any age) and 94.9% (40-50 years). Absolute risk for CIN2+ in HR-HPV positive and HPV16/18 positive individuals was 25.9 and 35.1, respectively. Relative risk for CIN2+ lesions was 5.1 for HPV16/18 positive versus HR-HPV negative, and 3.8 for HR-HPV positive versus HR-HPV negative women. Predictive values of CIN2+ positive were higher for HPV16/18 positive women (any age) than 12 other HPV positive-genotypes, and highest (50%) at 40-50 years. The HPV16/18 genotyping might prevent women (>40 years) at risk of high-grade CIN lesions from undergoing unnecessary colposcopy/overtreatment of nonprogressive lesions.

Potential impact of combined high- and low-risk human papillomavirus infection on the progression of cervical intraepithelial neoplasia 2.

AIM: Few studies have examined the effect of combined low-risk human papillomavirus (LR-HPV) and high-risk human papillomavirus (HR-HPV) infection on the progression of cervical intraepithelial neoplasia (CIN)2 to CIN3. This multi-institutional prospective cohort study investigated the risk of progression of CIN2 with various combinations of HR-HPV and LR-HPV infection. METHODS: Between January 2007 and May 2008, 122 women with CIN2 (aged 20-50 years) from 24 hospitals throughout Japan were enrolled in the study. Ninety-three women were analyzed after a 2-year follow-up with cytology, colposcopy, HR-HPV testing and HPV genotyping. Colposcopy-directed biopsy was performed at entry and the end of this study, or when disease progression was suspected. RESULTS: Among 93 women with CIN2, 87 (93.5%) had HR-HPV infection. Among these 87 cases, 24 (27.6%) progressed to CIN3 and 49 (56.3%) regressed. None of the six women with CIN2 without HR-HPV infection progressed. The progression rate was significantly lower in women with combined HR-HPV and LR-HPV infection (3/28, 10.7%) than in those with HR-HPV infection only (21/59, 35.6%; P = 0.016). Multivariate analyses showed that CIN2 progression in women with HR-HPV infection was negatively associated with LR-HPV co-infection (hazard ratio = 0.152; 95% confidence interval [CI] = 0.042-0.553). CIN2 regression was positively associated with LR-HPV co-infection (odds ratio = 4.553; 95% CI = 1.378-15.039). CONCLUSION: The risk of CIN2 progression is low in women with combined infection of HR-HPV and LR-HPV. The finding may be useful for management of women diagnosed with CIN2.

Prospective evaluation of the Amplicor HPV test for predicting progression of cervical intraepithelial neoplasia 2.

AIM: The aim of this study was to evaluate the clinical performance of the Amplicor HPV test, which detects 13 high-risk human papillomaviruses (HR-HPV), and to determine the association between consistent HR-HPV infection and progression of cervical intraepithelial neoplasia (CIN) 2 to CIN3. MATERIAL AND METHODS: This multi-institutional prospective study enrolled 122 women diagnosed with CIN2 by central pathological review. Subjects were tested at study entry and every 6 months over a 24-month period by cytology, Amplicor HPV test and colposcopy. Central pathological review was performed at the end of the study or if CIN progression was suspected. RESULTS: Ninety-three of the 122 participants completed all tests in the study and were included in the analysis. HR-HPV was detected in 87/93 (93.5%) participants at study entry. Twenty-four of the 87 HR-HPV-positive participants progressed to ≥CIN3, compared with none of the six participants who were HR-HPV-negative at study entry. The positive predictive value, negative predictive value, sensitivity and specificity of the Amplicor HPV test at study entry for predicting ≥CIN3 progression were 27.6%, 100%, 100% and 8.7%, respectively. Sixty-two participants were HR-HPV-positive from study entry through to study completion, 24 of whom progressed to ≥CIN3. None of 31 participants without continuous HR-HPV detection progressed to ≥CIN3. For continuous HR-HPV detection, the positive predictive value, negative predictive value, sensitivity and specificity of the Amplicor HPV test were 38.7%, 100%, 100% and 44.9%, respectively. CONCLUSIONS: All participants who progressed to ≥CIN3 were continuously HR-HPV-positive. The Amplicor HPV test thus demonstrated a good performance for predicting CIN3 progression.

Differences in parameters of the explosive grip force test between young and older women.

BACKGROUND: Decreases of strength and speed of muscle contraction are considered major causes of functional impairments in older people. However, the age-related changes in grip force-generating capacity are not clear. The purpose of this study was to measure the parameters of the explosive grip force test and to compare the differences between young and older women. METHODS: Thirty healthy young women (mean age: 22.3 years) and 27 healthy older women (mean age: 78.5 years) participated in this study. All participants performed the maximal explosive grip test three times. Data were recorded as a force-time curve, and the maximal rate of grip force development (max RGFD) and RGFD at intervals of 10 ms up to 250 ms from the onset of contraction were calculated. RESULTS: The majority of RGFDs of young women were higher than those of the older ones. The maximal grip strength, max RGFD, and max RGFD normalized by the maximal grip strength of older women were 28.3%, 52.4%, and 25.2% less than those of young women, respectively. RGFDs of the older women were not influenced by the maximal grip strength, whereas in young women, those in the late phase of explosive grip force generation showed a moderate correlation with the maximal grip strength. CONCLUSIONS: The present results showed a decrease of the output parameters of the explosive grip force test in older women. Evaluation of explosive grip force generation using RGFD may be used as an assessment tool, providing more detailed information on the grip function.

Ribosomal protein profiling by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for phylogenety-based subspecies resolution of Bifidobacterium longum.

The taxonomic positions of the subspecies of Bifidobacterium longum (B. longum subsp. longum, subsp. infantis, and subsp. suis) have been controversial. A current proposal is that the former two species "B. infantis" and "B. suis" be unified with B. longum and all three reclassified as three subspecies. To test this proposal, ribosomal protein profiling as observed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was applied to the classification of 17 strains of B. longum, including three subspecies. Among 41 different kinds of ribosomal proteins selected as biomarkers whose masses were calculated from their amino acid sequences, 31-41 ribosomal proteins were observed in sample strains with the same masses as the references. The high matching rate indicates high conservation of ribosomal proteins within the sample strains, and therefore strongly supports the unification of the former species. However, the masses of some ribosomal proteins varied within species. The phylogenetic tree constructed from the profiles of ribosomal proteins matched the references, showing a clear cluster of the subsp. longum and the subsp. infantis strains. This result supports the proposal to reclassify B. longum into subsp. longum and subsp. infantis. The subsp. suis strains formed an individual sub-cluster within the infantis cluster. However, their ribosomal proteins have both characters of longum and infantis types. This result suggests that the taxonomic position of the subsp. suis should be reconsidered.

Effects of propolis from different areas on mast cell degranulation and identification of the effective components in propolis.

Propolis is considered to down-regulate type I allergy, but the effective components of propolis remain unknown. In addition, propolis components vary depending on the area from which they are collected due to variations among wild plants in an area. Therefore, we compared the effects of water and ethanol extracts of propolis from Brazil and China on mast cell degranulation and cytokine production, thereby identifying effective components in propolis. The amount of released beta-hexosaminidase via high-affinity IgE receptor I (Fc epsilon RI) from rat basophilic leukemia (RBL-2H3) cells was used as an index of degranulation. All propolis extracts inhibited degranulation from antigen-stimulated RBL-2H3 cells, but the effective doses differed according to collection areas. The ethanol extract of Chinese propolis, which was the strongest inhibitor of mast cell degranulation, was divided into compounds using normal- and reversed-phase liquid chromatography. The isolated anti-allergic components were identified as chrysin, kaempferol and its derivative, and chrysin was revealed to inhibit IL-4 and MCP-1 production from antigen-stimulated RBL-2H3 cells. HPLC quantification also revealed the Brazilian propolis extract to contain only small amounts of these flavonoids, which suggested that variation in propolis components could affect anti-allergic properties.

Clinical significance of human papillomavirus genotype by linear array assay in Japanese women with uterine cervical lesions and type 16 physical status by in situ hybridization.

The aim of this study was to clarify the relation of human papillomavirus (HPV) genotypes and physical status in the cervical neoplasm of Japanese patients with the grade of the disease. Human papillomavirus genotype was detected using a linear array genotyping assay. Human papillomavirus status, diffuse or punctate signal pattern, was studied by biotynyl-tyramide-based in situ hybridization for positive cases of HPV-16. Human papillomavirus types 16, 52, 58, and 31, in descending order of frequency, were prevalent. The rates of HPV infection in patients with cervical intraepithelial neoplasia (CIN) or squamous cell carcinoma (SCC) were significantly higher than those in patients without cervical lesions. The frequency of HPV single infection in SCC was higher than that in CIN1 or CIN2. In an unspecified-risk HPV, types 66 and 70 were found in SCC and 62, 71, and 82 were detected in CIN3. The diffuse pattern was more frequent in CIN, and the punctate pattern was more frequent in SCC. Human papillomavirus types 16, 52, 58, and 31 were frequently detected in Japanese women with cervical neoplasias, and several unspecified-risk HPVs might be high-risk types. A single infection of HPV and a punctate signal pattern seemed to be closely correlated with cervical carcinogenesis.

Fad24 causes hyperplasia in adipose tissue and improves glucose metabolism.

We have previously reported that a novel gene, factor for adipocyte differentiation (fad) 24, promotes adipogenesis in vitro. To examine the role of fad24 in adipogenesis in vivo and the development of obesity, transgenic mice overexpressing fad24 were generated using mouse fad24 cDNA under the control of a chicken beta-actin promoter and cytomegalovirus enhancer. The comparison of the ability of fibroblasts from fad24 transgenic embryos to differentiate into adipocytes with that of fibroblasts from wild-type embryos revealed that fad24 overexpression promotes adipogenesis in embryonic fibroblasts. The weight and histology of white adipose tissues, and serum adipocytokine levels were compared between fad24 transgenic mice and wild-type mice, and we found that fad24 overexpression increased the number of smaller adipocytes, caused hyperplasia rather than hypertrophy in white adipose tissue and increased the serum adiponectin level in mice fed both normal chow and a high-fat diet. Glucose and insulin tolerance tests indicated that the activity for glucose metabolism is improved in fad24 transgenic mice fed normal chow in comparison with that in wild-type mice. Our findings suggest that fad24 is a positive regulator of adipogenesis in vivo. Moreover, the increase in the number of smaller adipocytes caused by the overexpression of fad24 appears to enhance glucose metabolic activity, perhaps by increasing the serum adiponectin level.

Effect of alachlor on hepatic cytochrome P450 enzymes in rats.

Alachlor ((2-chloro-N-methoxymethyl)-N-(2,6-diethylphenyl)acetamide) is a widely used preemergence herbicide which has been classified by the USEPA as a probable human carcinogen. The herbicide has been suggested to be metabolized by hepatic cytochrome P450 system. We examined the effects of alachlor on cytochrome P450 enzymes in rat liver microsomes. Rats were treated intraperitoneally with alachlor daily for 5 days, at doses of 25, 50 and 100 mg/kg. Among the cytochrome P450-dependent monooxygenase activities, 7-pentoxyresorufin O-depentylase, which is associated with CYP2B1, was dose-dependently increased by alachlor. The induction relative to control activity was 1.7-4.2-fold. The activities of CYP1A-dependent monooxygenases such as 7-ethoxy-resorufin O-deethylase and acetanilide 4-hydroxylase were also significantly increased by alachlor at doses of 50 and 100 mg/kg (1.7-2.1-fold). Furthermore, immunoblotting showed that alachlor significantly increased CYP2B1/2 and CYP1A1/2 protein levels by 4.2-6.3- and 1.8-fold, respectively. Although 7-ethoxycoumarin O-deethylase, bufuralol 1'-hydroxylase and 4-nitrophenol 2-hydroxylase activities were significantly increased by alachlor at higher doses (> or = 50 mg/kg), the induction ratios were less than 1.6-fold. The activities of other cytochrome P450-dependent monooxygenases, namely testosterone 7 alpha-hydroxylase, testosterone 2 alpha-hydroxylase, testosterone 6 beta-hydroxylase and lauric acid omega-hydroxylase, were not affected by alachlor at any dose. In addition, there was no significant change in the protein levels of CYP2C11/6, CYP2D1, CYP2E1, CYP3A2/1 and CYP4A1/2/3. These results suggest that alachlor selectively induces cytochrome P450 isoforms of the CYP1A and CYP2B subfamilies in rat liver microsomes, and that the expression of these isoforms is closely related to the toxicity of alachlor.