Molecular Diagnosis of Thiophanate-Methyl-Resistant Strains of Fusarium fujikuroi in Japan.

Fusarium fujikuroi is the pathogen of rice bakanae disease and is subclassified into gibberellin and fumonisin groups (G and F groups). Thiophanate-methyl (TM), a benzimidazole fungicide, has been used extensively to control F. fujikuroi. Previous investigation showed that F-group strains are TM sensitive (TMS), whereas most G-group strains are TM resistant (TMR) in Japan. The minimum inhibitory concentration in TMS strains was 1 to 10 µg ml-1, whereas that in TMR strains was >100 µg ml-1. E198K and F200Y mutations in ß2-tubulin were detected in TMR strains. A loop-mediated isothermal amplification-fluorescent loop primer method was developed for diagnosis of these mutations and applied to 37 TMR strains and 56 TMS strains. The results indicated that 100% of TMR strains were identified as having either the E198K mutation (41%) or the F200Y mutation (59%), whereas none of the TMS strains tested showed either mutation. We found one remarkable TMR strain in the F group that had an F200Y mutation. These results suggest that E198K and F200Y mutations in ß2-tubulin contribute to TM resistance in F. fujikuroi.

Physiological and Transcriptional Response of Xanthomonas oryzae pv. oryzae to Berberine, an Emerging Chemical Control.

Berberine, a botanical drug, has great ability to inhibit the growth of Xanthomonas oryzae pv. oryzae. However, the antibacterial mechanism of berberine against X. oryzae pv. oryzae remains poorly understood. In this study, we investigated the physiological and transcriptional response of X. oryzae pv. oryzae to berberine. When strain X. oryzae pv. oryzae GX13 was treated with berberine (10 µg/ml), the hypersensitive response in tobacco, virulence to rice, pathogen population in the rice xylem, production of extracellular polysaccharide (EPS), and activity of extracellular hydrolases decreased, but the levels of pyruvate and ATP increased. Moreover, biofilm formation was inhibited, and the cell membrane was damaged. Transcriptome sequencing analysis showed downregulated expression of gspD, gspE, and gspF, involved in the type II secretion system (T2SS); hrcC, hrcJ, hrcN, and others, involved in the type III secretion system (T3SS); gumB and gumC, associated with EPS; zapE, ftsQ, and zapA, associated with cell division; lpxH, lpxK, kdtA, and others, associated with the membrane; and pyk, pgk, and mdh, encoding pyruvate kinase, phosphoglycerate kinase, and malate dehydrogenase, respectively. Upregulated expression was observed for nuoA, nuoB, and nuoH, encoding the NADH dehydrogenase complex, and atpF, atpC, and atpB, encoding ATP synthase. An adenylate cyclase (CyaA) fusion assay showed that berberine affects type three effector protein secretion via the T3SS and reduces effector translocation in X. oryzae pv. oryzae. It is speculated that the negative growth and virulence phenotypes of berberine-treated X. oryzae pv. oryzae GX13 may involve differentially expressed genes associated with cytoarchitecture and energy metabolism, and these effects on primary cell function may further dampen virulence and result in differential expression of T3SS- and T2SS-related genes.

Kinetic Curve Type Assessment for Classification of Breast Lesions Using Dynamic Contrast-Enhanced MR Imaging.

OBJECTIVE: The aim of this study was to employ a kinetic model with dynamic contrast enhancement-magnetic resonance imaging to develop an approach that can efficiently distinguish malignant from benign lesions. MATERIALS AND METHODS: A total of 43 patients with 46 lesions who underwent breast dynamic contrast enhancement-magnetic resonance imaging were included in this retrospective study. The distribution of malignant to benign lesions was 31/15 based on histological results. This study integrated a single-compartment kinetic model and dynamic contrast enhancement-magnetic resonance imaging to generate a kinetic modeling curve for improving the accuracy of diagnosis of breast lesions. Kinetic modeling curves of all different lesions were analyzed by three experienced radiologists and classified into one of three given types. Receiver operating characteristic and Kappa statistics were used for the qualitative method. The findings of the three radiologists based on the time-signal intensity curve and the kinetic curve were compared. RESULTS: An average sensitivity of 82%, a specificity of 65%, an area under the receiver operating characteristic curve of 0.76, and a positive predictive value of 82% and negative predictive value of 63% was shown with the kinetic model (p = 0.017, 0.052, 0.068), as compared to an average sensitivity of 80%, a specificity of 55%, an area under the receiver operating characteristic of 0.69, and a positive predictive value of 79% and negative predictive value of 57% with the time-signal intensity curve method (p = 0.003, 0.004, 0.008). The diagnostic consistency of the three radiologists was shown by the κ-value, 0.857 (p<0.001) with the method based on the time-signal intensity curve and 0.826 (p<0.001) with the method of the kinetic model. CONCLUSIONS: According to the statistic results based on the 46 lesions, the kinetic modeling curve method showed higher sensitivity, specificity, positive and negative predictive values as compared with the time-signal intensity curve method in lesion classification.

Left ventricular ejection fraction estimation using mutual information on technetium-99m multiple-gated SPECT scans.

BACKGROUND: A new non-linear approach was applied to calculate the left ventricular ejection fraction (LVEF) using multigated acquisition (MUGA) images. METHODS: In this study, 50 patients originally for the estimation of the percentage of LVEF to monitor the effects of various cardiotoxic drugs in chemotherapy were retrospectively selected. All patients had both MUGA and echocardiography examinations (ECHO LVEF) at the same time. Mutual information (MI) theory was utilized to calculate the LVEF using MUGA imaging (MUGA MI). RESULTS: MUGA MI estimation was significantly different from MUGA LVEF and ECHO LVEF, respectively (p < 0.005). The higher repeatability for MUGA MI can be observed in the figure by the higher correlation coefficient for MUGA MI (r = 0.95) compared with that of MUGA LVEF (r = 0.80). Again, the reproducibility was better for MUGA MI (r = 0.90, 0.92) than MUGA LVEF (r = 0.77, 0.83). The higher correlation coefficients were obtained between proposed MUGA MI and ECHO LVEF compared to that between the conventional MUGA LVEF and ECHO LVEF. CONCLUSIONS: MUGA image with the aid of MI is promising to be more interchangeable LVEF to ECHO LVEF measurement as compared with the conventional approach on MUGA image.

Identification of Breast Cancer Using Integrated Information from MRI and Mammography.

OBJECTIVES: Integration of information from corresponding regions between the breast MRI and an X-ray mammogram could benefit the detection of breast cancer in clinical diagnosis. We aimed to provide a framework of registration from breast MRI to mammography and to evaluate the diagnosis using the combined information. MATERIALS AND METHODS: 43 patients with 46 lesions underwent both MRI and mammography scans, and the interval between the two examinations was around one month. The distribution of malignant to benign lesions was 31/46 based on histological results. Maximum intensity projection and thin-plate spline methods were applied for image registration for MRI to mammography. The diagnosis using integrated information was evaluated using results of histology as the reference. The assessment of annotations and statistical analysis were performed by the two radiologists. RESULTS: For the cranio-caudal view, the mean post-registration error between MRI and mammography was 2.2±1.9 mm. For the medio-lateral oblique view, the proposed approach performed even better with a mean error of 3.0±2.4 mm. In the diagnosis using MRI assessment with information of mammography, the sensitivity was 91.9±2.3% (29/31, 28/31), specificity 70.0±4.7% (11/15, 10/15), accuracy 84.8±3.1% (40/46, 38/46), positive predictive value 86.4±2.1% (29/33, 28/33) and negative predictive value 80.8±5.4% (11/13, 10/13). CONCLUSION: MRI with the aid of mammography shows potential improvements of sensitivity, specificity, accuracy, PPV and NPV in clinical breast cancer diagnosis compared to the use of MRI alone.

Significant association of caveolin-1 (CAV1) genotypes with upper urothelial tract cancer.

AIM: Upper urothelial tract cancer is unusually of high incidence in Taiwan and it is valuable to study the specificity of this disease in Taiwan and compare the corresponding findings with those of Western countries. In the literature, it has been reported that single nucleotide variation of caveolin-1 gene (CAV1) plays an important role in risk of several types of cancer, such as hepatoma, leukemia, nasopharyngeal carcinoma, oral, breast, bladder and prostate cancer, but we are not aware of any reports on upper urothelial tract cancer. The aim of this study was to evaluate the association of six polymorphic genotypes of CAV1 with upper urothelial tract cancer within a Taiwanese population. MATERIALS AND METHODS: A total of 218 patients with upper urothelial tract cancer and 580 healthy controls in central Taiwan were genotyped by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) for six CAV1 polymorphic genotypes, C521A (rs1997623), G14713A (rs3807987), G21985A (rs12672038), T28608A (rs3757733), T29107A (rs7804372), and G32124A (rs3807992), and their association with upper urothelial tract cancer susceptibility was examined. RESULTS: The distribution of genotypes of CAV1 rs3807987 and rs7804372 were significantly different between cancer patient and control groups (p=0.0188 and 0.0090, respectively), while those for CAV1 rs1997623, rs12672038, rs3757733 and rs3807992 were not significant (p>0.05). The haplotype analysis of the two polymorphic genotypes showed that compared with the GG/AT, and GG/AA haplotypes of CAV1 rs3807987/rs7804372, those carrying GG/TT, AG/TT and AA/TT variants have a significantly increased risk of upper urothelial tract cancer (odds ratio=1.61, 1.50 and 2.67, 95% confidence interval=1.05-2.47, 1.18-1.90, and 1.37-5.18, respectively). On the contrary, other haplotype variants conferred non-significant elevated risk. CONCLUSION: Our results suggest that individual and combined CAV1 rs3807987/rs7804372 genotypes are involved in predisposition to upper urothelial tract cancer in the Taiwanese population.

Association of cyclin D1 genotypes with nasopharyngeal carcinoma risk.

AIM: The cell cycle regulator cyclin D1 (CCND1) is a critical regulator of the G1/S phase transition and plays an important part in several tumor types. This study aimed at investigating the association of CCND1 with and examining the interaction among CCND1 genotype and individual smoking habit in nasopharyngeal carcinoma susceptibility. PATIENTS AND METHODS: A total of 352 native Taiwanese consisting of 176 cases and 176 controls were enrolled in this hospital-based study, and CCND1 A870G (rs9344) and C1722G (rs678653) genotyping were analyzed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and partially verified by direct sequencing. RESULTS: The results showed that there were significant differences between nasopharyngeal carcinoma and control groups in the distribution of the genotypic (p=0.0222) and allelic (p=0.0322) frequencies in the CCND1 A870G genotype. Individuals who carried at least one G allele (GG or AG) had a 0.71-fold lower risk of developing nasopharyngeal carcinoma compared to those who had the AA genotype (95% confidence interval=0.53-0.96). In addition, there is an obvious joint effect of CCND1 A870G genotype with smoking habit on nasopharyngeal carcinoma susceptibility. CONCLUSION: These findings support the conclusion that the cell cycle regulation may play a role in nasopharyngeal carcinoma development and that CCND1 A870G polymorphism maybe a useful biomarker for nasopharyngeal carcinoma progression.

Significant association of caveolin-1 (CAV1) genotypes with breast cancer in Taiwan.

AIM: Japanese and American groups reported that single nucleotide variation of caveolin-1 gene (CAV1) plays an important role in breast cancer risk. The aim of this study was to evaluate the association of six polymorphic genotypes of CAV1, which is reported to be overexpressed in tumors, with breast cancer within a Taiwanese population. PATIENTS AND METHODS: A total of 1232 patients with breast cancer and equal number of healthy controls in central Taiwan were genotyped via polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) and six polymorphic variants of CAV1 were analyzed for their association with breast cancer susceptibility. RESULTS: The distribution of genotypes of CAV1 G14713A and T29107A were significantly different between breast cancer and control groups (p=5.6×10(-5) and 1.9×10(-4), respectively), while those for CAV1 C239A, G21985A, T28608A and G32124A were not significant (p>0.05). The percentages of AG genotype of G14713A and TT genotype of T29107A are higher in the cancer group than in the control group. The two single nucleotide polymorphisms were chosen for haplotype analysis and the data showed that compared with GG/TT haplotype of CAV1 G14713A/T29107A, the GG/AT and GG/AA groups have a lower risk of breast cancer (odds ratio, OR=0.69, 95% confidence interval, CI=0.57-0.92). On the contrary, the AG/TT haplotype confers a higher risk of breast cancer (OR=1.50, 95% CI=1.14-2.12). CONCLUSION: Our results provide evidence for CAV1 genotypes being involved in predisposition to breast cancer. The association of the potential risk haplotype agrees well with a role of CAV1 genotype in breast cancer risk and the association with tumor progression needs further investigation.

Association of caveolin-1 genotypes with nasopharyngeal carcinoma susceptibility in Taiwan.

BACKGROUND: Caveolin-1 (Cav-1), which has been proposed as a candidate tumor suppressor, plays a regulatory role in several signaling pathways. High expression of Cav-1 in nasopharyngeal carcinoma (NPC) may enhance tumor cell migration and correlate with poor prognosis of the patients, while the genetic alterations of Cav-1 during nasopharyngeal carcinogenesis are still largely unknown. The aim of this study was to evaluate the association between NPC susceptibility and Cav-1 genotypes. PATIENTS AND METHODS: One hundred and seventy six patients with NPC and 176 age- and gender-matched healthy controls recruited in Taiwan were genotyped and analyzed by PCR-restriction fragment length polymorphism. RESULTS: There were significant differences between the NPC and control groups in the distributions of the genotypic (p=0.0019) and allelic frequencies (p=2.5 10(-4)) in the Cav-1 T29107A (rs7804372) polymorphism. CONCLUSION: In this first report of Cav-1 involvement in NPC the A allele of Cav-1 T29107A is found to be protective against the development of NPC and may be a novel useful genomic marker for early screening and prediction of NPC.