Anyplex II HPV28 detection and Anyplex II HPV HR detection assays are highly concordant with other commercial assays for detection of high-risk HPV genotypes in women with high grade cervical abnormalities.

PURPOSE: to evaluate the performance of Anyplex II HPV28 and HPV HR Detection assays against the EuroArray HPV, Cobas 4800 HPV (Cobas), HPV Amplicor (Amp), Linear Array HPV (LA) and Hybrid Capture 2 (HC2) in detection of high-risk HPV (HR-HPV) from liquid-based cervical cytology samples. METHODS: cervical specimens from 404 women undergoing management of high-grade cytological abnormality were evaluated by Anyplex II HPV28 and HPV HR Detection assays for detection of HR-HPV genotypes and prediction of histologically-confirmed cervical intraepithelial neoplasia grade 2 or higher (≥CIN2). The results were compared to EuroArray, HC2, Cobas, Amp, and LA. RESULTS: specimens were evaluated from 404 women with an average age of 30 years, including 336 with a histological diagnosis of ≥ CIN2 and 68 with ≤ CIN1. Concordance of HR-HPV detection between Anyplex II HPV28 and other genotyping assays was 94.79 % (κ = 0.84; EuroArray) and 97.27 % (κ = 0.91; LA); and between Anyplex II HPV HR and other HR-HPV detection assays was 86.35 % (κ = 0.62; HC2), 96.03 % (κ = 0.87; Cobas) and 96.77 % (κ = 0.89; Amp). Using HR-HPV detection for prediction of ≥ CIN2 by Anyplex II HPV28 and HPV HR, sensitivity (90.18, 95 % CI 86.48-93.14; 90.77, 95 % CI 87.16-93.65) and specificity (both 67.16, 95 % CI 54.60-78.15) were not significantly different to the other HPV assays tested, with one exception. Both Anyplex assays had significantly higher sensitivity than HC2 (p < 0.0001), with a specificity of 96 % (p > 0.05) of HC2 in this high-risk population. CONCLUSIONS: both Anyplex II HPV detection assays were concordant with other commercial assays for HR-HPV detection, with comparable sensitivity and specificity for ≥ CIN2 detection.

EUROarray human papillomavirus (HPV) assay is highly concordant with other commercial assays for detection of high-risk HPV genotypes in women with high grade cervical abnormalities.

The purpose of this study was to evaluate the performance of the EUROIMMUN EUROArray HPV genotyping assay against the Roche Cobas 4800, Roche HPV Amplicor, Roche Linear Array and Qiagen Hybrid Capture 2 assays in the detection of high-risk HPV (HR-HPV) from liquid based cervical cytology samples collected from women undergoing follow-up for abnormal cervical cytology results. Cervical specimens from 404 women undergoing management of high-grade cytological abnormality were evaluated by EUROarray HPV for detection of HR-HPV genotypes and prediction of histologically-confirmed cervical intraepithelial neoplasia grade 2 or higher (≥CIN2). The results were compared to Hybrid Capture 2, Cobas 4800 HPV, Amplicor and Linear Array HPV. Positivity for 14 HR-HPV types was 80.0 % for EUROarray (95 % CI; 75.7-83.8 %). Agreement (κ, 95 % CI) between the EUROarray and other HPV tests for detection of HR-HPV was good to very good [Hybrid Capture κ = 0.62 (0.54-0.71); Cobas κ = 0.81 (0.74-0.88); Amplicor κ = 0.68 (0.60-0.77); Linear Array κ = 0.77 (0.70-0.85)]. For detection of HR-HPV, agreement with EUROarray was 87.90 % (Hybrid Capture), 93.58 % (Cobas), 92.84 % (Amplicor) and 92.59 % (Linear Array). Detection of HR-HPV was not significantly different between EUROarray and any other test (p < 0.001). EUROarray was concordant with other assays evaluated for detection of high-risk HPV and showed sensitivity and specificity for detection of ≥ CIN2 of 86 % and 71 %, respectively.

The risk of preterm birth following treatment for precancerous changes in the cervix: a systematic review and meta-analysis.

BACKGROUND: Studies investigating the association between treatment for precancerous changes in the cervix and risk of preterm birth have used a variety of comparison groups. OBJECTIVES: To investigate whether treatment for precancerous changes in the cervix is associated with preterm birth (<37 weeks) and to examine the impact of the type of comparison group on estimates of risk. SEARCH STRATEGY: PubMed, Embase and CENTRAL were searched for studies pubished between 1950 and 2009. SELECTION CRITERIA: Eligible studies were those that reported preterm birth outcomes for excisional and ablative treatments separately and included a comparison group. DATA COLLECTION AND ANALYSIS: Pooled relative risks (RR) and 95% confidence intervals were computed using a random effects model. MAIN RESULTS: Thirty eligible studies were located. Excisional treatment was associated with an increased odds of preterm birth, when compared with an external (RR 2.19, 95% CI 1.93-2.49) or internal (RR 1.96, 95% CI 1.46-2.64) comparison group. In comparison with women who were assessed but not treated, the risk estimate was smaller (RR 1.25, 95% CI 0.98-1.58). Ablative treatment was associated with an increased risk of preterm birth when an external comparison group (RR 1.47, 95% CI 1.24-1.74) but not an internal comparison group (RR 1.24, 95% CI 0.73-2.10) or untreated comparison group (RR 1.03, 95% CI 0.90-1.18) was used. AUTHORS' CONCLUSIONS: Excisional treatment was associated with a significantly increased risk of preterm birth. It provides new evidence that some types of ablative treatment may also be associated with a small increased risk. The type of comparison group used is an important consideration when comparing the outcomes of studies.

Progestogen treatment options for early endometrial cancer.

OBJECTIVE: Premenopausal women with early endometrial cancer may wish to maintain their fertility, and for some patients non-surgical treatment options may be attractive. We have examined our own experience with such patients, as there are limited published data so far to support clear guidelines in this area. DESIGN: Retrospective analysis of a case series. SETTING: Case series from a specialist gynaecological oncology unit in a major tertiary referral hospital. SAMPLE: Sixteen patients receiving progestogen therapy for stage-1 endometrial cancer. METHODS: We reviewed our experience of all patients receiving progestogen therapy for stage-1 endometrial cancer, and we particularly examined their cancer-free outcome and fertility potential. MAIN OUTCOME MEASURES: Response to treatment, duration of response, and subsequent pregnancies. RESULTS: Of the 16 patients investigated, four received an oral progestogen, five received the levonorgestrel-releasing intrauterine system (Mirena), and seven received both forms of treatment. Ten patients (63%) responded to treatment, with a median time to response of 5.5 months. Six patients did not respond to treatment, but all were either early in treatment or opted for surgical management before the average time of response. No patient who responded had a later recurrence. The mean total follow-up time was 27 months (range 3-134 months), with no patient deaths. Three patients had successful pregnancies, with one patient having two children. CONCLUSIONS: This form of treatment appears to be a realistic treatment option in selected patients in the closely supervised environment of a specialist gynaecological oncology unit.

Detection of cervical intraepithelial neoplasia in vivo using confocal endomicroscopy.

OBJECTIVE: A high resolution optical imaging device may offer a clinically useful adjunct to colposcopy for the diagnosis and assessment of cervical precancer. This study describes the clinical evaluation of a miniaturised confocal endomicroscope for the quantitative and qualitative assessment of cervical intraepithelial neoplasia (CIN) in vivo. DESIGN: A descriptive study (n = 25) was performed to assess the usability of confocal endomicroscopy to image the cervix. A prospective study (n = 15) then evaluated the diagnostic accuracy of the technique. SETTING AND POPULATION: Patients undergoing colposcopy for treatment of CIN1-CIN3 were examined using confocal endomicroscopy. METHODS: A 5% solution of acetic acid was used to enhance the colposcopic features of the atypical region. Normal and abnormal regions of the cervix were then imaged following topical application of a fluorescent dye (acriflavine). MAIN OUTCOME MEASURES: Confocal images were analysed to develop a scoring system to grade different levels of CIN. Microscopic features were correlated with histology from biopsy. RESULTS: Confocal endomicroscopy enabled microscopic imaging of cellular and subcellular structures in vivo at colposcopy. Imaging at increasing depth showed morphological features including dermal papillae, endocervical glands and the squamo-columnar junction. CIN was characterised by an increase in nuclear density, size and cellular atypia. The sensitivity for detection of CIN was 97%. The specificity for predicting the grade of abnormality was 80% for normal-CIN1 and 93% for CIN2-CIN3. CONCLUSIONS: Confocal endomicroscopy is a sensitive imaging tool for detection and assessment of CIN. The technique enables in vivo imaging of cervical histology and the potential for 'see-and-treat' workflows.

An immunohistochemical perspective of PPAR beta and one of its putative targets PDK1 in normal ovaries, benign and malignant ovarian tumours.

Peroxisome proliferator-activated receptor beta (PPAR beta) is a member of the nuclear hormone receptor family and is a ligand-activated transcription factor with few known molecular targets including 3-phosphoinositide-dependent protein kinase 1(PDK1). In view of the association of PPAR beta and PDK1 with cancer, we have examined the expression of PPAR beta and PDK1 in normal ovaries and different histological grades of ovarian tumours. Normal ovaries, benign, borderline, grades 1, 2 and 3 ovarian tumours of serous, muciuous, endometrioid, clear cell and mixed subtypes were analysed by immunohistochemistry for PPAR beta and PDK1 expression. All normal ovarian tissues, benign, borderline and grade 1 tumours showed PPAR beta staining localised in the epithelium and stroma. Staining was predominantly nuclear, but some degree of cytoplasmic staining was also evident. Approximately 20% of grades 2 and 3 tumours lacked PPAR beta staining, whereas the rest displayed some degree of nuclear and cytoplasmic staining of the scattered epithelium and stroma. The extent of epithelial and stromal PPAR beta staining was significantly different among the normal and the histological grades of tumours (chi(2)=59.25, d.f.=25, P<0.001; chi(2)=64.48, d.f.=25, P<0.001). Significantly different staining of PPAR beta was observed in the epithelium and stroma of benign and borderline tumours compared with grades 1, 2 and 3 tumours (chi(2)=11.28, d.f.=4, P<0.05; chi(2)=16.15, d.f.=4, P<0.005). In contrast, PDK1 immunostaining was absent in 9 out of 10 normal ovaries. Weak staining for PDK1 was observed in one normal ovary and 40% of benign ovarian tumours. All borderline and malignant ovarian tumours showed positive cytoplasmic and membrane PDK1 staining. Staining of PDK1 was confined to the epithelium and the blood vessels, and no apparent staining of the stroma was evident. Significantly different PDK1 staining was observed between the benign/borderline and malignant ovarian tumours (chi(2)=22.45, d.f.=5, P<0.001). In some borderline and high-grade tumours, staining of the reactive stroma was also evident. Our results suggest that unlike the colon, the endometrial, head and neck carcinomas, overexpression of PPAR beta does not occur in ovarian tumours. However, overexpression of PDK1 was evident in borderline and low- to high-grade ovarian tumours and is consistent with its known role in tumorigenesis.

The value of synovial cytokine expression in predicting the clinical response to TNF antagonist therapy (infliximab).

OBJECTIVES: Clinical response to TNF-alpha blockade in the treatment of RA is heterogeneous. The study aims were to determine whether pre-treatment synovial cytokine expression predicted infliximab response and whether synovial changes after therapy correlated with response. METHODS: Fifty-one patients had arthroscopic biopsies of the knee joint prior to infliximab (3 mg/kg) treatment. Synovial tissue cell numbers (CD68 and CD3 positive) and cytokine expression (TNF-alpha, lymphotoxin-alpha, IL-1alpha, -beta and receptor antagonist, and IL-6) pre-treatment was assessed using semi-quantitative immunohistochemistry. Changes in these parameters were assessed 16 weeks after infliximab in 32 patients who underwent repeat arthroscopic biopsy. RESULTS: Of the total patients, 47% (n = 24) achieved an ACR20 response; 53% (n = 27) did not. Baseline synovial TNF-alpha, IL-1alpha and -beta expression did not differ between the two groups. No differences in baseline TNF-alpha levels were observed with ACR levels of response (ACR20 and ACR50/70 groups). Post-treatment biopsies (17 ACR responders, 15 ACR non-responders) revealed significant reductions in sub-lining layer TNF-alpha expression in both response and non-response groups with significant reduction in vascularity and membrane proliferation scores. The worst ACR non-responders (<20% CRP suppression) demonstrated no reduction in any of the parameters. CONCLUSION: Pre-treatment synovial TNF-alpha or IL-1 expression does not predict TNF blockade response. Both ACR response and non-response was associated with reduction in synovial TNF-alpha-level expression. Suppression in TNF-alpha levels was not observed in the worst non-responders. The improvements (including in vascularity), independent of ACR clinical response, are compatible with the reduced structural damage documented in all groups of patients independent of response.

Prognostic factors for recurrence in patients with FIGO stage I and II, intermediate or high risk endometrial cancer.

BACKGROUND: The aim of this study was to determine predictors for loco-regional or distant recurrence of disease in a subgroup of intermediate or high risk stage I and II endometrial cancer. METHODS: A retrospective analysis of 295 patients with histopathological stage I and II, intermediate or high risk endometrial cancer is reported. The following factors were studied: stage, grade, age, histologic diagnosis, lymphadenectomy, lymphovascular space invasion, and adjuvant radiotherapy. The Log-Rank test was used for statistical analyses and the Kaplan-Meyer method was used for time-to-event analysis. Multivariate analysis was also performed. RESULTS: Thirty-four (11.5%) patients developed a recurrence; 20 (59%) developed loco-regional recurrence, and 14 (41%) developed distant recurrence. In 20 women (59%), recurrence appeared within 3 years of surgery, and the actuarial survival at 3 years after recurrence was 29%. Multivariate analysis showed that for recurrence, age >60 years was a significant unfavourable prognostic factor (p < 0.05). CONCLUSIONS: We found low rates of recurrence in patients with early stage intermediate or high risk endometrial cancer. Only age was identified as an independent significant predictor for recurrence.

An improved tripod amphiphile for membrane protein solubilization.

Intrinsic membrane proteins represent a large fraction of the proteins produced by living organisms and perform many crucial functions. Structural and functional characterization of membrane proteins generally requires that they be extracted from the native lipid bilayer and solubilized with a small synthetic amphiphile, for example, a detergent. We describe the development of a small molecule with a distinctive amphiphilic architecture, a "tripod amphiphile," that solubilizes both bacteriorhodopsin (BR) and bovine rhodopsin (Rho). The polar portion of this amphiphile contains an amide and an amine-oxide; small variations in this polar segment are found to have profound effects on protein solubilization properties. The optimal tripod amphiphile extracts both BR and Rho from the native membrane environments and maintains each protein in a monomeric native-like form for several weeks after delipidation. Tripod amphiphiles are designed to display greater conformational rigidity than conventional detergents, with the long-range goal of promoting membrane protein crystallization. The results reported here represent an important step toward that ultimate goal.

Variability in the fatty acid composition of wax esters from vernix caseosa and its possible relation to sebaceous gland activity.

Wax ester/sterol ester (WE/SE) ratios were measured in samples of vernix caseosa lipid obtained from 21 full-term infants. The mean WE/SE ratio was twice as high in males as in females, suggesting higher average fetal sebum production rates in males, but individual values varied widely and there was considerable overlap between the sexes. Wax esters (a class of lipid produced only by the sebaceous glands) were isolated from 6 of the 21 samples of vernix caseosa lipid and analyzed for fatty acid composition. The percentages of iso and anteiso branched saturated fatty acids and of delta 9-type monounsaturated fatty acids were found to be correlated negatively with the WE/SE ratios of the original lipid samples. Values for the percentages of the major delta 9-type monounsaturates, plotted as a function of WE/SE ratios, fell near a curve of the shape which would be expected if a limited quantity of delta 9-type monounsaturates were available for sebum synthesis, and were diluted with varying amounts of delta 6-type monounsaturates, the amount of the delta 6-type monounsaturates being proportional to the amount of wax esters synthesized by the sebaceous glands. The results suggested that the rate of sebum production may be the sole determinant of the percentage of delta 9-type monounsaturates in sebaceous wax esters and a partial determinant of the percentage of iso and anteiso branched saturates.

Cell free synthesis of Toxoplasma gondii antigens.

Functionally active poly(A)-containing mRNA was isolated from tachyzoites of the RH strain of Toxoplasma gondii. The T. gondii mRNA was capable of directing the synthesis of proteins in a wheat germ in vitro translation system, but not in a cell free system derived from rabbit reticulocyte lysate. Efficient translation in the wheat germ system required spermine and exogenous tRNA. Amino acid incorporation was maximal at 110 mM K+ and 1.8 mM Mg2+. Tachyzoite antigens synthesized in vitro were immunoprecipitated with T. gondii antibodies from rabbits, mice and humans. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of immunoprecipitated polypeptides yielded patterns that differed according to antibody source, but all T. gondii antibody preparations reacted with a translation product with an apparent molecular weight of 24 000.

Factors influencing serum CA 125 levels in normal women.

Serum CA 125 levels were studied in 2544 healthy women. Menopausal status and a history of hysterectomy were both highly significant influences on the CA 125 level (P less than .0001 for both factors). A history of hormone replacement therapy was also highly significant in reducing CA 125 levels (P = .002) in postmenopausal women. Other factors such as parity, history of unilateral oophorectomy or oral contraceptive usage, or family history of ovarian cancer were not significant influences when analysis of covariance was performed. When premenopausal women were assessed separately, age was significant (P = .04). The number of years since menopause did not influence CA 125 levels. Both hysterectomy and menopausal status have a clear effect on serum CA 125 levels and must be considered if serum CA 125 is to be used as a screening test.

Response of a mucinous ovarian tumor of borderline malignancy to human chorionic gonadotropin.

Plasma and urinary steroid hormones were measured before and after an injection of human chorionic gonadotropin (hCG) to a postmenopausal woman with a mucinous ovarian tumor of borderline malignancy. Hormones were also measured in blood from a vein draining the tumor, and circulating gonadotropins and plasma and urinary steroids were measured before and after tumor removal. Baseline levels of plasma progesterone (P), androstenedione (delta 4 A), and estradiol (E2), and urinary estrogens and pregnanediol were high; they increased dramatically in response to hCG and fell after tumor removal. A less striking increase in testosterone, dihydrotestosterone, dehydroepiandrosterone (DHEA), and DHEA sulfate was noted after hCG injection. A gradient existed between tumor vein and peripheral vein levels of P, 17 alpha-hydroxyprogesterone, delta 4 A, E2, DHEA, and cortisol. Plasma follicle-stimulating and luteinizing hormones initially low but rose to the postmenopausal range after surgery. These results indicate the presence of delta 4 and delta 5 androstene pathways within the tumor. The responsiveness of the tumor to hCG provides further evidence that hCG may be the endogenous stimulus to steroid hormone production by epithelial ovarian tumors.

Functioning ovarian tumors in postmenopausal women.

Estrogen excretion was higher than normal (more than 9 microgram/24 hr) in 40 of 80 postmenopausal women with benign and malignant epithelial tumors of the ovary, including 8 metastatic tumors. High estrogen excretion was noted in 19 of 27 (70.4%) patients with mucinous tumors in only 1 of 25 (4%) patients with serous tumors (P less than .001). Endometrioid and metastatic tumors were also noted to be frequently associated with high estrogen excretion. In the high estrogen excretion group 27 of 37 (73%) patients showed stromal luteinization and/or condensation in the tumor, but in the normal estrogen excretion group these changes were seen in only 6 of 38 (15.8%) patients (P less than .001). A possible mechanism of stromal stimulation in these tumors is discussed. Although there was often evidence of estrogen activity in the endometrium and the tubal epithelium in the high estrogen excretion group, the correlation between these epithelia and estrogen excretion was imperfect. Postoperative estrogen excretion remained elevated in 3 of 16 patients, of whom 1 was obese and 2 had residual tumor. This study emphasizes the importance of identifying epithelial tumors of the ovary as functioning or endocrine tumors.

The prognostic importance of steroid receptors in endometrial carcinoma.

Three hundred forty-nine cases of primary endometrial carcinoma (endometrioid, adenosquamous, and clear-cell) were studied to investigate the relative prognostic importance of age, menopausal status, stage, histology, myometrial invasion, and estrogen and progesterone receptor content. Excluding menopausal status, all of these variables had a significant relationship to overall survival in a univariate analysis. Using a Cox multivariate regression analysis, stage, age, and an estrogen receptor value of more than 70 fmol/mg protein, combined with a progesterone receptor value of more than 30 fmol/mg protein, were independently associated with survival. The results demonstrate that for maximum prognostic information, both estrogen and progesterone content of tumors should be measured. Maximum prognostic information is obtained by using cutoff levels that are much higher than those traditionally accepted. This has particular relevance for patient stratification in clinical trials investigating receptor information and response to adjuvant or therapeutic treatment.

Hormonal treatment of endometrial cancer.

This article summarizes the endocrine background of women with endometrial cancer at both peripheral and tissue levels, and the current status of clinical trials of hormonal, cytotoxic, and combination regimens. Because significant advances in systemic therapy are required to improve the prognosis of endometrial cancer, recommendations for future clinical investigations will be based on these recent biologic observations.

Estimation of platelet size by measurement of intracellular water space using an oil technique.

The intracellular water space of human platelets has been measured after equilibration with tritiated water and then separating these cells by centrifugation through phthalate oil of density 1.042. The mean intracellular water space of platelets in citrated plasma was 0.52 +/- 0.09 microliter/10(8) cells for 19 normal subjects. The gravimetric water content of platelets was 784 +/- 4 mg water/g cells. From these values the mean platelet volume was calculated to be 6.2 fl which agrees closely with values based on Coulter size distribution and thrombocytocrit. Gel filtration alters platelets such that a mean 19% of the platelets could not be centrifuged through phthalate oils of density 1.031 or 1.042. The measurement of tritiated water space of platelets centrifuged from their own plasma through oil provides a simple and reliable estimate of the mean platelet size.

How do you diagnose rheumatoid arthritis early?

There are difficulties in making an accurate diagnosis of rheumatoid arthritis in its early stages, a principal problem being the fact that its most defining feature is chronicity, which, by definition, takes time to identify. There is substantial evidence that patients with rheumatoid arthritis should be treated early, and the majority are now commenced on therapy when first diagnosed. A logical implication of the early therapy approach is that treatment before rheumatoid arthritis is fully developed may have even greater benefits. This requires patients who are likely to have a persistent, more severe disease to be identified and treated effectively in the very earliest stages. This requires a system of the early specialist referral of suitable patients and the use of effective predictors of patient outcome. This text discusses the early arthritis clinic approach and the current evidence base available for use in constructing management guidelines for such patients.

Window of opportunity in early rheumatoid arthritis: possibility of altering the disease process with early intervention.

Therapeutic strategies for the treatment of rheumatoid arthritis (RA) have changed significantly in the last decade. The emphasis is now on early intervention with the aim of preventing disability and irreversible damage. Advocates of early intervention would suggest an alteration in the disease process, not just debulking of inflammatory disease. The data would at least support attenuation of the disease process with aggressive early therapy. Further research is required to elucidate the scientific mechanisms involved and their impact on the pathological progress of RA.

Collision tumor: serous adenocarcinoma and steroid cell tumor of the ovary.

A "collision" tumor between a serous papillary adenocarcinoma and a steroid cell tumor of the ovary is described. No similar combination has been reported in the literature. The steroid cell component secreted testosterone, resulted in considerable virilization of the patient, and appears to have preceded the carcinoma by several years. It remains problematical whether the androgenic milieu may have predisposed to the development of the second, malignant, tumor.