A simple and robust flow injection on-line microdialysis system for multivessel drug dissolution testing is described. Microdialysis probes were used for sampling from the dissolution media. A stopped-flow dialysis mode with a 50-s stopped-flow period followed by 10-s injection at a perfusion rate of 2.8 ml min(-1) for each probe was used to achieve high resolution of dissolution events using relatively simple equipment and operation. The precisions obtained for simultaneous monitoring of dissolution profiles for six tablets were all better than 0.9%(RSD n=80) and the overall sampling frequency of the system was 360 and 60 h(-1) for each test vessel. The dissolution profiles of isoniazid fast-release tablet from three sources were determined to demonstrate the performance of the system.

A sequential injection system for the determination of mercury by vapor generation atomic absorption spectrometry (VGAAS) using tetrahydoborate reductant was developed, characterized by prevention of sample and reagent mixing in the holding coil using small air segments and initiation of the vapor generation in a flow-through gas-liquid separator. Extremely small volumes of reductant of 15-30 mul (0.2-1.0% NaBH(4)) and sample acidity as low as 0.05 mol l(-1) HCl were sufficient for achieving performance similar to flow injection (FI) VGAAS systems. A sample throughput of 90 h(-1) was achieved with 400 mul samples with a precision of 2.0% RSD at 10 mug l(-1)Hg, and a detection limit of 0.1 mug l(-1)(3sigma). Reagent consumption was reduced by a factor of 25 in comparison to the FI-VGAAS system. Good agreement with the certified value was obtained for the determination of mercury in seawater in a standard reference sample.

Fitness costs associated with resistance genes expressed in the absence of insecticides affect the evolution of insecticide resistance and the outcome of resistance management programmes. However, measurements of fitness costs may not be straightforward as they vary with environmental conditions. The diamondback moth, Plutella xylostella L.(Lepidoptera: Plutellidae), has developed resistance to spinosad, the first insecticide of the Naturalyte class, after only a few years of field application of this product. In this study, we compared the performance of two homogenous strains of P. xylostella, one susceptible (SS) and the other resistant (RR) to spinosad at an unfavourable, low natural temperature regime, a favourable median-fluctuating temperature regime and an unfavourable high-fluctuating temperature regime. The RR strain showed only marginal fitness cost at the median temperature regime. At the low temperature regime, the RR strain failed to produce any viable offspring, while the SS strain achieved positive population growth. At the high temperature regime, the RR strain showed a 33% decrease in intrinsic rate of increase compared to the SS strain. The results demonstrate that fitness costs of resistance to spinosad are temperature-dependent, increasing in scale at unfavourably low and high temperatures; costs were particularly high at low temperatures. Suggestions for designing effective management programmes are made to delay or avoid development of resistance to spinosad by P. xylostella under different temperature conditions.

Coccidiosis of chickens caused by protozoan parasites of the genus Eimeria (Coccidia: Eimeriidae) is an enteric disease that results in great economic losses throughout the world, including Taiwan. Using polymerase chain reaction (PCR) with primers specific for the second internal transcribed spacer (ITS-2) of ribosomal DNA (rDNA), three species of Eimeria, E. tenella, E. maxima, and E. acervulina have been successfully characterised from chickens in Taiwan. The sizes of PCR products from various isolates representing these three species were between 370 and 580 base pairs (bp). After cloning and sequencing of the PCR products, high nucleotide sequence identity (96.8-100%) was observed within a species. In addition, ITS-2 nucleotide sequences for E. tenella had higher homology (98.5-99.3%) than E. maxima (81.6-96.5%) when compared with appropriate sequences deposited in GenBank. To our knowledge, this is the first report of a 412-bp ITS-2 sequence for E. acervulina from chickens.

BACKGROUND: Intrinsic radiosensitivity using the clonogenic assay and the cell surviving fraction at 2 Gy (SF2) has been shown to be an independent prognostic factor for patient response to radiotherapy in carcinoma of the cervix. The clonogenic assay has significant shortcomings, making it unsuitable for routine clinical use. The ATP cell viability assay (ATP-CVA) has been shown to have a high tumor evaluability rate, technical simplicity, and reproducibility in chemosensitivity testing. AIMS: This study compares the ATP-CVA with the clonogenic assay in the in vitro radiosensitivity testing of cervical cancer cell lines. Correlation of in vitro radiosensitivity and in vivo patient response was also determined. METHODS: Five cervical carcinoma cell lines (SiHa, HeLa, Caski, C-33A, and C4-1) were tested using the ATP-CVA and the clonogenic assay. Survival curves were plotted and the mean SF2 values obtained by the two different assay methods were compared using ANOVA to see if there were significant differences. Mean SF2 values obtained from 27 cervical cancers were compared with clinical outcomes. RESULTS: The SF2 values for the cell lines ranged from 0.28 to 0.67 when tested using the ATP-CVA. Using the clonogenic assay, the SF2 values ranged from 0.27 to 0.70. ANOVA with Bonferroni pairwise multiple comparison showed no significant difference between the mean SF2 values for the individual cell lines between the two assay methods. Twenty-three cervical cancer samples (85%) were evaluable for SF2 using ATP-CVA. The mean SF2 values of patients who had locoregional failure were significantly higher than those who achieved local control (P <0.01). CONCLUSIONS: Testing intrinsic radiosensitivity using the surviving fraction at 2 Gy (SF2) is comparable using the two assay methods of ATP-CVA and clonogenic assay. The ATP-CVA should be further investigated in the testing of intrinsic radiosensitivity in patients with cervical cancer.

The chalcid, Oomyzus sokolowskii Kurdjumov has been recorded in many parts of the world as a major larval-pupal, gregarious endoparasitoid of the diamondback moth, Plutella xylostella (Linnaeus), a serious pest of brassica vegetable crops worldwide. This study investigated intraspecific variation between two populations of O. sokiolowskii, one from Cape Verde Islands, West Africa and the other from Hangzhou, China. In all crosses and backcrosses between the two geographical populations, the numbers of progeny and sex ratio of progeny were similar to those obtained within each of the populations, demonstrating complete reproductive compatibility between the two populations. The two populations showed similar responses to temperature with respect to development time and survival of immature stages. Observations on the interactions between the two O. sokolowskii populations and Cotesia plutellae (Kurdjumov), another major parasitoid of P. xylostella, showed that neither population could achieve successful parasitism of P. xylostella larvae already parasitized by C. plutellae. However, both O. sokolowskii populations could achieve hyperparasitism by ovipositing into a mid-late stage larva of C. plutellae developing inside the primary host. Contrary to earlier reports, no evidence of intraspecific variations in ability to hyperparasitize between these two populations of O. sokolowskii was found.

A previous study showed E-cadherin expression was lost in some cervical cancer cell lines and tumours. This study was designed to clarify the significance of DNA methylation in silencing E-cadherin expression. We examined promoter methylation of E-cadherin in five cervical cancer cell lines and 20 cervical cancer tissues using methylation-specific PCR (MSP) and bisulphite DNA sequencing. The correlation of E-cadherin methylation and expression together with methyltransferase (DNMT1) were further studied. We found that hypermethylation of E-cadherin was involved in five cervical cancer cell lines and 40%(8/20) of cervical cancer tissues. E-cadherin protein was lost in 6/8 (75%) samples and 3/5 (60%) cell lines with promoter methylation. E-cadherin methylation was significantly correlated with increased DNMT1. Using an antisense DNMT1 oligo to transfect into SiHa HeLa C33A cell line, E-cadherin protein was re-expressed. We concluded that loss of E-cadherin expression was in part correlated with DNA methylation and DNMT1 expression in cervical cancer.

OBJECTIVE: The aim of this study was to evaluate the thymidine phosphorylase (TP) activity of tumor cells and serum as a marker in patients with ovarian cancer. METHODS: The activity of TP was measured by the spectrophotometric method in the cytosol of ovarian tumor tissues from 47 patients after surgery, and in the presurgery serum from the same patients. Ten women with normal ovaries, treated surgically due to nononcological reasons served as a control. Microvessel density (MD) was evaluated in tumor using immunohistochemical methods. A relationship between TP activity and MD and clinicopathologic features was investigated. RESULTS: A significantly higher TP activity was stated both in malignant tumor and serum specimens from ovarian cancer patients when compared to the control. A positive correlation between the enzyme activity in the serum and neoplasmatic tissue was found. Neoangiogenesis is higher in ovarian cancer when compared to the group of borderline malignancy tumors but a reverse correlation between MD and TP activity in malignant tumors was observed. TP activity is significantly higher in more advanced neoplasmatic disease (FIGO III and IV) although no correlation between TP activity and grading or histopathological type of ovarian tumor was observed. CONCLUSION: Thymidine phosphorylase activity might be useful in diagnostic characterization of ovarian cancer.

Serum tumor markers are useful in diagnosis and follow-up for patients with gynecological malignancy or breast cancer. In epithelial ovarian cancer, CA125 has been identified as the most sensitive marker. Unfortunately, CA125 detection in the serum of patients with minimal malignant tumor has not been possible. Many nonmalignant conditions including endometriosis, menstruation and massive ascites may elevate the CA125, and almost 50% of patients with clear cell adenocarcinoma do not show CA125 elevated above 100 U/ml. To improve sensitivity and specificity in the diagnosis of ovarian cancer, the use of multiple tumor markers and the simultaneous use of image diagnosis should be employed. The value of tumor markers in the screening for cervical cancer and endometrial cancer has received little attention. However, the utility of serum SCC as a marker for monitoring cervical squamous cell carcinoma has been established. Since hCG is produced by gestational trophoblastic neoplasia and is a sensitive marker of trophoblastic cells in the body, patients with choriocarcinoma or invasive mole must be followed closely for this parameter. The improvement of the hCG detection technique has reduced the mortality rate from trophoblastic neoplasia. In breast cancer, many markers including CEA and CA15-3 are used, and they are reported to be useful as markers for monitoring.

CA 125 is the most reliable serum marker for ovarian carcinoma. Whereas its role in the screening of the malignancy is controversial, serum CA 125 assay is very useful for both the differential diagnosis of ovarian masses, particularly in postmenopause, and the monitoring of the response to chemotherapy and follow-up of patients with histologically proven ovarian carcinoma. Tumor-associated antigens other than CA 125, such as CA 19.9, CA 15.3 and TAG.72, firstly identified in gastro-intestinal or breast malignancies, have been detected also in tissue and serum samples from patients with ovarian carcinoma. In particular CA19.9 offers the advantage of high sensitivity for mucinous histotype, which often fails to express CA 125. Serum CA 125 correlates with the clinical course of disease better than the other antigens, and in patients with positive CA 125 assay at diagnosis the concomitant evaluation of CA 19.9 or CA 72.4 or CA 15.3 does not offer any additional benefit for monitoring ovarian carcinoma. Conversely, the serial measurements of these other antigens may represent an interesting biochemical tool for the management of patients with negative CA 125 assay. Serum alphaFP and betaHCG are very useful in the preoperative evaluation and management of nondysgerminomatous ovarian germ cell tumors, whereas elevated serum inhibin levels can be detected in patients with granulosa cell tumors of the ovary. As for endometrial carcinoma, preoperative serum CA 125 levels correlate with stage, depth of myometrial invasion, histologic grade, cervical invasion, peritoneal cytology, lymph node status and clinical outcome. Moreover, serial CA 125 assay is a good indicator of disease activity and a useful biochemical tool for post-treatment surveillance of patients with endometrial carcinoma. SCC is the most reliable serum marker for squamous cell cervical carcinoma, and in patients with this malignancy pretreatment SCC levels are related to tumor stage, tumor size, depth of cervical invasion, lymph-vascular space involvement, lymph node status and clinical outcome. Serial SCC measurements parallel the response to radiotherapy and chemotherapy as well as the clinical course of disease after the completion of treatment. Serum CYFRA 21.1 seems to be less sensitive than serum SCC for squamous cell cervical carcinoma. Elevated CA 125 levels can be often detected in patients with cervical adenocarcinoma. The future for tumor marker research is represented by the emerging technologies of transcriptional profiling and proteomics.

BACKGROUND: In this study the clinical and technical performance of the CA125- detecting Bayer ACS:OV immunoluminometric serum assay was compared with three other well-established CA125-detecting assays. METHODS: A total of 1112 serum samples was included in this evaluation: 462 from apparently healthy women, 153 from patients with benign ovarian disease, 163 from patients with malignant ovarian disease, 10 from patients with borderline ovarian malignancies and 78 samples from 12 ovarian cancer patients during monitoring of disease. Serum samples from women with malignant endometrial (n = 68) and colon (n = 32) diseases were also included. Moreover, serum samples from women with benign uterine disease and endometriosis (n = 136) plus 10 serum samples from men (n = 7) and women (n = 3) with human anti-mouse antibodies (HAMA) after immunoscintigraphy were included. All samples were tested in duplicate with the Bayer ACS:OV, the Centocor CA125 II, the Abbott IMx CA125 and the Roche (formerly Boehringer Mannheim) Enzymun-Test CA125 II assays. RESULTS: The clinical performance of the Bayer ACS:OV assay, assessed in various patient groups, was similar to that of the two other automated assays. In serum from patients with benign diseases the highest values were found in patients with benign ovarian tumours. In the ovarian cancer patients followed during the course of disease we found similar marker patterns with all four assays. In contrast to the Roche Enzymun-CA125 II assay and to a lesser extent the Centocor CA125 II assay, the Bayer ACS:OV assay was less sensitive to interference from HAMA. CONCLUSION: The Bayer ACS:OV assay is a precise and reliable test for the quantification of CA125 in serum.

AIM. Our aim was to confirm that preoperative CA 125 serum level can be useful for discrimination between benign and malignant masses in the pelvis. METHODS. Preoperative CA 125 serum level was analyzed retrospectively in 121 patients who had surgery because of a malignant ovarian tumor and in 91 patients with benign masses in the pelvis. The cutoff serum level CA 125 between benign and malignant masses in the pelvis was 35 and 65 IU/ml. RESULTS. Of those patients with a malignant ovarian tumor, 65.3% had menopause whereas only 31.5% of those with a benign tumor did so. The average age of the patients with a malignant tumor was 54.2 years and of those with a benign tumor 46.8 years. The preoperative CA 125 serum level was higher than 35 IU/ml in 80.2% and higher than 65 IU/ml in 72.7% of all analyzed patients with a malignant tumor, whereas it was 23.9% and 9.8% respectively in patients with a benign mass. In early stage ovarian cancer disease (borderline stage, I/II) the preoperative CA 125 serum level was higher than 35 IU/ml in 67.8% and in 52.5% higher than 65 IU/ml. In advanced stages (III/IV), it was higher than 35 and 65 IU/ml in 96.1%. After therapy the CA 125 serum level dropped below 35 IU/ml in 70.8% and after three chemotherapy courses in 78.1%. A CA 125 level less than 35 IU/ml was achieved by therapy in 84.2% patients with an early stage disease (I/II) and in 62.1% in advanced stages (III/IV). The calculated sensitivity was 80.2% and negative 74.5%(CA 125 higher than 35 IU/ml) and 72.7%, 90.2%, 90.7%, 71.6% respectively (CA 125 higher than 65 IU/ml). CONCLUSION. Preoperative determination of CA 125 is a very useful method to discriminate between benign and malignant masses in the pelvis.

OBJECTIVE: To study the diagnostic significance and clinical effect of the detection of serum sialic acid (SA) in three kinds of malignant tumor. METHODS: The specimens (172) were divided into the healthy control group (100), lung cancer group (31), leukemia group (28) and rectual cancer group (17), and then the serum SA concentration in those specimens was analyzed and statistic analysis was conducted. RESULTS: The SA levels of the malignant tumor groups before and after the treatment were significantly higher than those of the healthy control group (P < 0.05). The positive rates of serum SA in the lung cancer group, leukemia group and rectual group were 84%, 79% and 71%. CONCLUSION: The SA concentration was significantly increased in the malignant patients so SA can be used as an indication of cancer, and it can be widely used in the diagnosis and inspection of malignant tumor.

The aim of the study was to investigate whether a combination of serum CA-125, CA-72.4; CA-19.9 and VEGF levels along with several currently used gray-scale criteria and pulsed and semiquantitative color Doppler blood flow assessment can be useful in preoperative discrimination of ovarian tumors. Ultrasound examinations were performed between 1994 and 1999 with the use of transvaginal probes 5-6.5-7.0 MHz and transabdominal 3.5 MHz probe (B&K Medical 3535 and 2002 ADI, Denmark). Preoperative serum CA-125 and CA-19.9 (Roche, Switzerland) and CA-72.4 (Centocor, USA) were measured with EIA II immunoassays. VEGF (R&D Systems, USA) serum levels were measured by microplate immunoenzymatic method. Retrospective study included 4,876 women referred for sonography of pelvic structures. For final analysis 451 patients with persistent adnexal tumors were available. In all these women age, menopausal status, tumor size, volume and morphology as well as blood flow indices (PI, RI, PSV) and semiquantitative color assessment were noted. In women who were not operated the cysts were followed for 10-12 weeks until resolution which was confirmed by repeated sonography. Multiple regression analysis was used to determine which of the independent variables had prognostic significance in the constructed predictive model. ROC curves were plotted and areas under ROC for each test were calculated and compared. 286 women were operated and in 118 patients their tumors regressed. The sensitivity for Doppler indices was between 85%(RI), 67%(PSV) and 66%(PI). For the subjective color assessment sensitivity was 87%. 294 women had their CA-125 serum levels available. For CA-19.9, CA-72.4 and VEGF the corresponding numbers were 109, 92 and 116 patients, respectively. Optimal cutoff level for CA-125 was 67.5 U/ml and this produced diagnostic accuracy of 69.4%. For CA-19.9 the best predictive value was 13.4 U/ml (accuracy of 74%) and for the cut-off CA-72.4 level of 4.1 U/ml the accuracy of the test was 77.8%. Multiple regression analysis revealed that only RI measurements and subjective color assessment had significant influence on the constructed predictive model. The best cut-off value of regression index "Z" was obtained following ROC curves construction for sensitivity (true positive rate) and 1-specificity (false positive rate). In all 116 patients who had Doppler indices with both CA-125 and VEGF levels measured the highest accuracy was associated with "Z"= 1.45 in postmenopausal women. The prognostic model proposed in this study can be log transformed and further used in the estimation of the true probability of malignancy of a given mass in the new set of prospectively studied patients with ovarian tumors. Color and pulsed Doppler can improve preoperative diagnosis of adnexal tumors when compared to transvaginal sonography alone or tumor markers assessment. The proposed semiquantitative evaluation of tumor vascularity increases the predictive value in terms of sensitivity and specificity. VEGF serum concentration was not useful in the preoperative discrimination of malignant and benign ovarian tumors.