Early stage detection and screening of ovarian cancer: A research opportunity and significant challenge for biosensor technology.

Women diagnosed with late-stage ovarian cancer suffer a very high rate of mortality. Accordingly, it is imperative to detect and diagnose the disease as early as possible in its development. Achievement of this aim implies relatively large scale screening of women at an age of clinically significance through assay of biomarkers for disease present in blood or serum. Biosensor detection offers an attractive technology for the automated detection of such species. Among several biomarkers that have been identified that are present in patients with ovarian cancer, the only one that is commonly tested for in clinical use is cancer antigen 125, which is considered to be poor biomarker for the disease. Here, we describe alternative biomarkers which overcome many of the problems associated with cancer antigen 125 such as increased sensitivity and specificity especially in the early stages of the disease and which could be employed successfully in a biosensor format. In particular, we discuss the chemistry of probes for the biomarkers, heat shock protein 10 and lysophosphatidic acid. The challenges presented by the fabrication of biosensor devices for the detection of the cancer, and the limited number of biosensors that have been developed for this purpose are discussed.

Markers of implantation in ectopic and high-risk early eutopic pregnancies.

INTRODUCTION: This study focused on the assessment of HSP-10, HSP-27 and PSG-11 which are one of the first detectable serum pregnancy proteins. Contrary to ultrasound imaging, biochemical methods allow to clarify the pathogenesis and pathomechanism of high-risk pregnancies, fetal anomalies, and abnormal fetal implantation. Early serum concentration estimation of HSP-10, HSP-27 and PSG-11 may be very useful not only in prognosis of pregnancies of unknown localization (PUL), but also as markers of ectopic pregnancies. OBJECTIVES: The aim of the study was to evaluate the expression of HSP-10, HSP-27, PSG-11 implantation proteins in ectopic and eutopic pregnancies, and their mutual correlations. PATIENTS AND METHODS: The study involved 42 healthy women who were hospitalized, due to symptoms of imminent miscarriage, risk of spontaneous abortion, or the diagnosis of an ectopic pregnancy. The subjects were subdivided into two equal groups of 21 women who consented to participate in this clinical trial. Biochemical assays were performed involving PSG-11, HSP-27, and HSP-10 serum concentration. RESULTS: Serum concentration levels of HSP-10, HSP-27, and PSG-11 were significantly higher in pregnancies at risk of spontaneous abortion as compared to ectopic pregnancies. CONCLUSIONS: The results of the study indicate high value of PSG-11, HSP-27 and HSP-10 serum concentrations as predictors of correct implantation site. This may be very useful in prognosis of pregnancies of unknown localization (PUL) and early conservative/surgical ectopic pregnancies treatment if necessary to preserve maximum fertility.

Evaluation of human chaperonin 10 and high-sensitivity C-reactive protein levels of infertile women who underwent ovulation induction and intra-uterine insemination.

INTRODUCTION: The implantation of embryo is one of the crucial steps of a successful pregnancy. The foetus should be protected from maternal immune system, for the appropriate implantation and modification in maternal immunity is crucial. We investigated high-sensitivity C-reactive protein (hs CRP), which is an indicator of low-grade inflammation and Cp10 that has immunosuppressant and growth-promoting capabilities at embryo levels in ovulation induction and intra-uterine insemination (IUI)applied in infertile women. The ovulation induction was maintained by clomiphene citrate or gonadotropins for 42 infertile patients. After successful ovulation induction, IUI was carried out. The blood samples were taken 2 and 8 days after IUI to evaluate Cp10 and hs CRP levels. The pregnant and non-pregnant groups' results were analyzed. The Cp10 levels 8 days after IUI were higher in pregnant group, whereas there was no difference for the 2 days after levels between pregnant and non-pregnant group. The hs CRP levels were similar for both 2nd and 8th days when we compared pregnant and non-pregnant groups. The Cp10 levels increased from day 2 to day 8 in pregnant group. In contrast, the Cp10 levels decreased in non-pregnant group. The change in hs CRP levels from day 2 to day 8 was not significant in pregnant and non-pregnant groups. The Cp10 levels were higher in early phases of fertilisation and elevated through the preceding days of conception in pregnant patients, while it decreased in non-pregnant patients with failed cycles.

Early pregnancy sex steroids and maternal breast cancer: a nested case-control study.

Pregnancy, parity, and circulating steroid hormone levels are associated with risk of breast cancer, but little is known about hormone concentrations during pregnancy and subsequent breast cancer risk. We evaluated early pregnancy (<140 days gestation) serum estradiol, estrone, progesterone, and testosterone and breast cancer risk in a nested case-control study in the Finnish Maternity Cohort. The cohort includes 98% of pregnancies registered in Finland since 1983. Individuals with samples collected in the first pregnancy leading to a live birth were eligible. Breast cancer cases (n = 1,199) were identified through linkage with the Finnish Cancer Registry; 2,281 matched controls were selected using incidence density sampling. ORs were calculated using conditional logistic regression. Hormone concentrations were not associated with breast cancer overall. Estradiol was positively associated with risk of breast cancer diagnosed age <40 [4th vs. 1st quartile OR 1.60 (1.07-2.39); Ptrend = 0.01], and inversely associated with breast cancer diagnosed at age ≥40 [4th vs. 1st quartile OR 0.71 (0.51-1.00); Ptrend = 0.02]. Elevated concentrations of the steroid hormones were associated with increased risk of estrogen receptor (ER)- and progesterone receptor (PR)-negative tumors in women age <40 at diagnosis. We observed no association between steroid hormones and ER(+)/PR(+) disease. These data suggest a positive association between high concentrations of early pregnancy steroid hormones and risk of ER(-)/PR(-) breast cancer in women diagnosed age <40, and an inverse association for overall breast cancer diagnosed age ≥40. Further research on pregnancy hormones and risk of steroid receptor-negative cancers is needed to further characterize this association.

Circulating heat shock protein 60 levels are elevated in HIV patients and are reduced by anti-retroviral therapy.

Circulating heat shock protein 60 (Hsp60) and heat shock protein 10 (Hsp10) have been associated with pro- and anti-inflammatory activity, respectively. To determine whether these heat shock proteins might be associated with the immune activation seen in HIV-infected patients, the plasma levels of Hsp60 and Hsp10 were determined in a cohort of 20 HIV-infected patients before and after effective combination anti-retroviral therapy (cART). We show for the first time that circulating Hsp60 levels are elevated in HIV-infected patients, with levels significantly reduced after cART, but still higher than those in HIV-negative individuals. Hsp60 levels correlated significantly with viral load, CD4 counts, and circulating soluble CD14 and lipopolysaccharide levels. No differences or correlations were seen for Hsp10 levels. Elevated circulating Hsp60 may contribute to the immune dysfunction and non-AIDS clinical events seen in HIV-infected patients.

Measurement of EPF for detection of cow pregnancy using rosette inhibition test.

Early embryonic death of calves due to sub-fertility in cows is of great economic concern to dairy industry. Early pregnancy factor (EPF) is a secretory protein with pregnancy associated immunosuppressive properties. Rosette inhibition test (RIT) was used to detect EPF in inseminated dairy cows. Blood samples were collected at two intervals, 1-3 and 5-7 days after insemination from 23 inseminated and 18 non-inseminated control cows for RIT and pregnancy diagnosis performed between 42 and 45 days on palpation. The study indicates that RIT (P<0.05) has the potential to distinguish pregnant from non-pregnant dairy cows in the first week of pregnancy.

HSP-10 in ovarian cancer: expression and suppression of T-cell signaling.

OBJECTIVE: Suppressed T-cell activation is a hallmark of advanced ovarian cancer. Studies in pregnancy have demonstrated similar T-cell dysfunction mediated, at least in part, by HSP10, identified as "early pregnancy factor." This pilot study addresses the presence of HSP10 in the circulation of ovarian cancer patients and assesses its role in suppressing CD3-zeta. METHODS: Sera were obtained from ovarian cancer patients (n = 10) and age-matched noncancer-bearing female controls (n = 9). HSP10 presence was determined semiquantitatively by Western immunoblotting in sera, ascites, and ovarian tumor cell conditioned media. The consequences of HSP10 on CD3-zeta suppression were defined using a Jurkat cell bioassay, using unfractionated patient sera, sera with HSP10 removed by immunoprecipitation and the immunoprecipitate. RESULTS: HSP10 was detected in both sera and ascites of patients with ovarian cancer; however, it was not detectable in controls. HSP10 was also detected in the culture media of ovarian tumor cells. Sera containing HSP10 suppressed T-cell CD3-zeta expression, which correlated with HSP10 levels (r2 = 0.839). When HSP10 was removed from the sera, the ability to suppress CD3-zeta was diminished and the immunoprecipitated material was capable of suppressing CD3-zeta. CONCLUSIONS: HSP10 appears to be produced and released from ovarian tumor cells and is detectable in the peripheral blood and ascites of patients. This circulating HSP10 appears to suppress T-cell expression of CD3-zeta, a key component of T-cell activation. Our findings indicate that, as in pregnancy, production and release of HSP10 may be a critical factor in the suppression of T-cell activation, allowing the tumor to escape immune surveillance.

Expression of the chaperonin 10 gene during zebrafish development.

We have isolated a cDNA encoding chaperonin 10 (cpn10) from the zebrafish. Using northern, western, and in situ hybridization analysis, we observed that the cpn10 gene is expressed uniformly and ubiquitously throughout embryonic development of the zebrafish. Upregulation of cpn10 expression was observed following exposure of zebrafish embryos to a heat shock of 1 hour at 37 degrees C compared to control embryos raised at 27 degrees C. The extracellular form of Cpn10 called early pregnancy factor (EPF), found in the serum of pregnant mammals, was not detected in the serum of either male or female zebrafish. These expression studies suggest that Cpn10 plays a general role in zebrafish development as well as being consistent with the hypothesis that EPF is involved in the embryo implantation process in mammals.

A study of early pregnancy factor activity in the sera of women with trophoblastic tumor.

PROBLEM: To detect whether or not the early pregnancy factor (EPF)-like activity, or chaperonin 10, could be in the sera of patients with trophoblastic tumor in order to find another more efficient means to diagnose this kind of tumor. METHOD OF STUDY: The rosette inhibition assay was used to detect EPF-like activity in 216 sera, collected from patients with gestational trophoblastic tumor, including 47 sera of patients with choriocarcinoma, 68 sera of patients bearing invasive mole, and 101 sera of patients with vesicular mole. RESULTS: The accuracy of diagnosing malignant trophoblastic tumor by detecting EPF-like activity is 91.3% (105/115), with a false positive rate of 14.58% and a false negative rate of 4.8% by this method. Furthermore, the rosette inhibition titer (RIT) values have significant difference (P < 0.001) between the sera in patients with malignant trophoblastic tumor before treatment and those after treatment. CONCLUSIONS: This study demonstrated that diagnosis of malignant trophoblastic tumor could be made with an accuracy of 91.3% by detecting EPF-like activity and that EPF-like activity could be used as an indicator to distinguish benign from malignant trophoblastic tumor.

Early pregnancy factor: an extracellular chaperonin 10 homologue.

Early pregnancy factor (EPF) has been identified as a homologue of chaperonin 10 (cpn10) with immunosuppressive and growth factor properties. As a homologue of cpn10, it belongs to the heat shock family of proteins (hsp) but, unlike other members of this family, EPF is detected extracellularly. Early pregnancy factor was first discovered in pregnancy serum by the rosette inhibition test, and the novelty of its discovery was that its presence could diagnose pregnancy within 6-24 h of a fertile mating. As well as being a monitor of the presence of a viable embryo, it is necessary for embryonic survival. In this capacity it acts as both an immunosuppressant and growth factor. Early pregnancy factor is also a product of proliferating primary and neoplastic cells and functions as an autocrine growth factor both in vivo and in vitro. It has a modifying effect on the outcome of experimental autoimmune encephalomyelitis, an animal model of multiple sclerosis. Early pregnancy factor is considered to be one of the major factors involved in the modification of multiple sclerosis observed during pregnancy.