Judith L. Luborsky

Histopathology of ovarian tumors in laying hens: a preclinical model of human ovarian cancer.

The high mortality rate due to ovarian cancer (OVCA) is attributed to the lack of an effective early detection method. Because of the nonspecificity of symptoms at early stage, most of the OVCA cases are detected at late stages. This makes the access to women with early-stage disease problematic and presents a barrier to development and validation of tests for detection of early stage of OVCA in humans. Animal models are used to elucidate disease etiologies and pathogenesis that are difficult to study in humans. Laying hen is the only available animal that develops OVCA spontaneously; however, detailed information on ovarian tumor histology is not available. The goal of this study was to determine the histological features of malignant ovarian tumors in laying hens. A total of 155 young and old (1-5 years of age) laying hens (Gallus domesticus) were selected randomly and evaluated grossly and microscopically for the presence of ovarian tumors. Histological classification of tumors with their stages and grades was determined with reference to those for humans. Similar to humans, all 4 types including serous, endometrioid, mucinous, and clear cell or mixed carcinomas were observed in hen ovarian tumors. Some early neoplastic as well as putative ovarian lesions were also observed. Similarities in histology, metastasis, and stages of hen OVCA to those of humans demonstrate the feasibility of the hen model for additional delineation of the mechanism underlying ovarian carcinogenesis, preclinical testing of new agents for the prevention, and therapy of this disease.

A population-based longitudinal study of cognitive functioning in the menopausal transition

Background: No longitudinal studies have tracked cognitive performance through the menopausal transition and thus the impact of the transition on cognition, independent of aging, is not known. The authors hypothesized that a decline in cognitive functioning occurs as women progress through the menopausal transition, independent of age, educational level, family income, ethnicity, and baseline self-perceived health. Method: The authors began a population-based, longitudinal study in January 1996 with yearly follow-up interviews. This report includes follow-up through November 2001. The authors randomly selected African American and white women from a census of two contiguous Chicago communities. After screening for eligibility (age 42 to 52 years, premenopausal or early perimenopausal, no exogenous hormone use in the past 3 months, and no hysterectomy), 868 agreed to participate. Women who became pregnant, had a hysterectomy, or began using hormones were censored from that time onward. This study reports on 803 women for whom cognitive assessments were available. The authors assessed working memory (Digit Span Backward) and perceptual speed (Symbol Digit Modalities Test). Results: Contrary to the hypothesis, the authors found small but significant increases over time during the premenopausal and perimenopausal phases. This trend was not accounted for by chronological age, education, family income, ethnicity, or baseline self-perceived health. Conclusions: Transition through menopause is not accompanied by a decline in working memory and perceptual speed.

Musculoskeletal pain and menopausal status.

ObjectivesThe authors examined whether self-reported menopausal status is associated with musculoskeletal pain in a multiethnic population of community-dwelling middle-aged women after considering sociodemographics, medical factors, smoking, depression, and body mass index using a cross-sectional study design. MethodsParticipants were 2218 women from the Study of Womens Health Across the Nation assessed at the time of their third annual follow-up exam. Two dependent variables were derived from a factor analysis of survey questions about pain. These 2 outcomes were Aches and Pains, derived from 5 of 6 pain symptom questions and Consultation for Low Back Pain, derived from 1 question. ResultsPrevalence of aches and pains was high, with 1 in 6 women reporting daily symptoms. Compared with premenopausal women, those who were early perimenopausal (P=0.002), late perimenopausal (P=0.002), or postmenopausal (P<0.0001) reported significantly more aches and pains in age-adjusted analysis. With complete risk factor adjustment, postmenopausal women still reported significantly greater pain symptoms (P=0.03) than did premenopausal women. Menopausal status was marginally related to consulting a healthcare provider for back pain. DiscussionThis study demonstrates an association between pain and self-reported menopausal status, with postmenopausal women experiencing greater pain symptoms than premenopausal women.

Autoantigens in ovarian autoimmunity associated with unexplained infertility and premature ovarian failure

OBJECTIVE To identify ovarian autoantigens associated with ovarian autoantibodies. DESIGN Hypothesis-generating prospective study. SETTING Urban infertility referral centers and academic research institution. PATIENT(S) Seventy-four patients with infertility, 19 patients with premature ovarian failure (POF), and 16 healthy control women. INTERVENTION(S) None. MAIN OUTCOME MEASURE(S) Identification of autoantigens. RESULT(S) To identify major antigens for ovarian autoimmunity, sera from 74 women with unexplained infertility were screened for ovarian autoantibodies (AOAs) by immunoassay and one-dimensional Western blot. The majority of sera had immunoreactions at 50-56 kDa. Six representative positive infertility sera were used to identify antigens between 40 and 60 kD by two-dimensional Western blot and mass spectrometry. Antigens included aldehyde (retinal) dehydrogenases (ALDH1A1, ALDH1A2, and ALDH7A1), protein disulfide isomerase A3, vimentin, α-enolase, phosphoglycerate dehydrogenase, and selenium-binding protein 1 (SBP1). Sixty percent (24 out of 40) of infertility and POF sera were positive for recombinant ALDH1A1, SBP1, or enolase; 80.7% (21 out of 26) of AOA-positive sera had antibodies to one or more of the three antigens, and only 7% (1 out of 14) of AOA-negative sera had antibodies to recombinant proteins. CONCLUSION(S) ALDH1A1 and SBP1 are unique to ovarian autoimmunity associated with infertility and POF, and may provide the basis for specific tests to identify patients with ovarian autoimmunity.

Factors related to declining luteal function in women during the menopausal transition.

CONTEXT Reproductive hormones are incompletely characterized during the menopause transition (MT). HYPOTHESIS Increased anovulation and decreased progesterone accompany progress through the MT. DESIGN The Daily Hormone Study (DHS) of the Study of Womens Health Across the Nation (SWAN) included 848 women aged 43-53 yr at baseline who collected daily urine for one cycle or up to 50 d annually for 3 yr. MAIN OUTCOME MEASURES LH, FSH, estrone conjugates, and pregnanediol glucuronide levels were assessed. Cycles were classified by presumed luteal (ovulatory) status and bleeding. Hormones were related to time in study, age, menopausal status, and selected variables. RESULTS Ovulatory-appearing cycles declined from 80.9% at baseline to 64.7% by the third assessment (H3). Cycles presumed anovulatory and not ending with bleeding by 50 d (anovulatory/nonbleeding) increased from 8.4 to 24% by H3 and were associated with progress to early perimenopause [odds ratio (OR) = 2.66; confidence interval (CI) = 1.17-6.04] or late perimenopause (OR = 56.21; CI = 18.79-168.12; P < 0.0001), African-American ethnicity (OR = 1.91; CI = 1.06-3.43), and less than high school education (OR = 3.51; CI = 1.62-7.62). Anovulatory cycles ending with bleeding remained at about 10% from baseline to H3; compared with ovulatory cycles, they were associated with obesity (OR = 4.68; CI = 1.33-16.52) and more than high school education (OR = 2.12; CI = 1.22-3.69; P = 0.02). Serum estradiol in both the highest and lowest categories was associated with anovulatory/nonbleeding collections. Pregnanediol glucuronide decreased 6.6% for each year on study. Insulin sensitivity measures did not relate strongly to menstrual cycle hormones. CONCLUSIONS Anovulation without bleeding represents progression of the MT. A small but detectable decrease in luteal progesterone excretion occurs as women progress through the MT.

Anti-tumor antibodies in ovarian cancer.

Anti‐tumor antibodies have potential as cancer biomarkers. There is relatively limited identification of anti‐tumor antibodies in response to ovarian cancer, compared with studies for other cancers. There is also very limited information on the prevalence of anti‐tumor antibodies among ovarian cancer patients. Although most anti‐tumor antibodies react with antigens common to both tumor and normal tissue, the anti‐tumor response tends to be confined to individuals with ovarian cancer, similar to other cancers. Antibodies to HOXA7, a differentiation antigen, have the highest reported prevalence in ovarian cancer (67%). Antibodies to other ubiquitous antigens including NY‐ESO‐1, Ep‐CAM (epithelial cell adhesion molecule), HSP‐90 (heat shock protein 90), and mutated p53 have been identified in ovarian cancer. Anti‐tumor antibody specificity reflects the heterogeneity of antigen expression in tumors. Tests based on panels of a combination of anti‐tumor antibodies may be more predictive for ovarian cancer, as no single specificity accounts for ovarian tumors. In addition to characterization of anti‐tumor antibodies as diagnostic markers, study of anti‐tumor antibodies is likely to provide insights into mechanisms of tumor development. There is evidence of antibodies to tumor antigens and of activated T cells, suggesting immune recognition of tumor antigens occurred. Nonetheless, as tumors are not ‘rejected’, it is likely that there are alterations in the immune system. The basis for tumor growth in the face of immune activity remains to be determined.

Differential expression of aldehyde dehydrogenase 1a1 (ALDH1) in normal ovary and serous ovarian tumors.

BackgroundWe showed there are specific ALDH1 autoantibodies in ovarian autoimmune disease and ovarian cancer, suggesting a role for ALDH1 in ovarian pathology. However, there is little information on the ovarian expression of ALDH1. Therefore, we compared ALDH1 expression in normal ovary and benign and malignant ovarian tumors to determine if ALDH1 expression is altered in ovarian cancer. Since there is also recent interest in ALDH1 as a cancer stem cell (CSC) marker, we assessed co-expression of ALDH1 with CSC markers in order to determine if ALDH1 is a potential CSC marker in ovarian cancer.MethodsmRNA and protein expression were compared in normal human ovary and serous ovarian tumors using quantitative Reverse-Transcriptase PCR, Western blot (WB) and semi-quantitative immunohistochemistry (IHC). ALDH1 enzyme activity was confirmed in primary ovarian cells by flow cytometry (FC) using ALDEFLUOR assay.ResultsALDH1 mRNA expression was significantly reduced (p < 0.01; n = 5) in malignant tumors compared to normal ovaries and benign tumors. The proportion of ALDH1+ cells was significantly lower in malignant tumors (17.1 ± 7.61%; n = 5) compared to normal ovaries (37.4 ± 5.4%; p < 0.01; n = 5) and benign tumors (31.03 ± 6.68%; p < 0.05; n = 5). ALDH1+ cells occurred in the stroma and surface epithelium in normal ovary and benign tumors, although surface epithelial expression varied more in benign tumors. Localization of ALDH1 was heterogeneous in malignant tumor cells and little ALDH1 expression occurred in poorly differentiated malignant tumors. In benign tumors the distribution of ALDH1 had features of both normal ovary and malignant tumors. ALDH1 protein expression assessed by IHC, WB and FC was positively correlated (p < 0.01). ALDH1 did not appear to be co-expressed with the CSC markers CD44, CD117 and CD133 by IHC.ConclusionsTotal ALDH1 expression is significantly reduced in malignant ovarian tumors while it is relatively unchanged in benign tumors compared to normal ovary. Thus, ALDH1 expression in the ovary does not appear to be similar to breast, lung or colon cancer suggesting possible functional differences in these cancers.SignificanceThese observations suggest that reduced ALDH1 expression is associated with malignant transformation in ovarian cancer and provides a basis for further study of the mechanism of ALDH1 in this process.

Selenium-Binding Protein 1 expression in ovaries and ovarian tumors in the laying hen, a spontaneous model of human ovarian cancer

OBJECTIVE Reduced Selenium-Binding Protein 1 (SELENBP1) expression was recently shown in multiple cancers. There is little information on the expression and function of SELENBP1 in cancer progression. In order to develop a better understanding of the role of SELENBP1 in ovarian cancer, our objective was to determine if SELENBP1 is expressed in the normal ovaries and ovarian tumors in the egg-laying hen, a spontaneous model of human ovarian cancer. METHODS SPB1 mRNA expression in normal ovary (n=20) and ovarian tumors (n=23) was evaluated by RT-PCR. Relative levels of mRNA were compared by quantitative RT-PCR (qRT-PCR) in selected samples. SELENBP1 protein expression was evaluated by 1D Western blot and immunohistochemistry with a commercial anti-human SELENBP1 antibody. RESULTS SELENBP1 mRNA and protein was expressed in 100% of normal and ovarian tumors and qRT-PCR confirmed decreased mRNA expression in 80% of ovarian tumors. SELENBP1 was primarily localized in surface epithelial cells of normal ovaries. In ovaries containing early tumor lesions, SELENBP1 expression was reduced in the surface epithelium near the tumor and was expressed in tumor cells, while more distant regions with normal histology retained SELENBP1 expression in the surface epithelium. CONCLUSIONS We have shown for the first time that SELENBP1 is expressed in both normal ovaries and ovarian tumors in the hen and that SELENBP1 expression is altered in the vicinity of the tumor. Furthermore, SELENBP1 expression in normal ovarian surface epithelium and in ovarian tumors parallels that previously reported for ovarian cancer in women.

Interpersonal violence, PTSD, and inflammation: potential psychogenic pathways to higher C-reactive protein levels.

Interpersonal violence (IPV) is major public health concern with wide-ranging sequelae including depression, posttraumatic stress disorder (PTSD), and possible alterations of immune and inflammation processes. There is a need to identify the psycho-biological pathways through which IPV may translate to altered inflammatory processes since both PTSD and inflammation are associated with serious physical health conditions such as obesity, diabetes, and cardiovascular disease. This study investigated the relationships between IPV, psychological distress, and the inflammatory marker C-reactive protein (CRP), in a sample of 139 urban women who have a high likelihood for having experienced IPV. Participants were recruited from an outpatient gynecology clinic to complete self-report measures about their IPV histories and psychological symptoms, as well as to have their blood sampled using a finger stick. Results indicated that exposure to IPV predicted the presence of probable depression and PTSD diagnoses. Individuals who experience clinical levels of PTSD exhibited higher CRP levels, and this relationship held after adjusting for comorbid depression. Correlational analyses suggested that reexperiencing symptoms may explain the link between PTSD diagnosis and higher levels of CRP. Follow-up path analytic models provided good fit to the overall data, and indicated that the relationship between probable PTSD status and CRP is not explained by higher BMI. Overall, these findings call for increased attention to the role of PTSD in explaining links between trauma and diminished health.

Detection of Tumor-Associated Neoangiogenesis by Doppler Ultrasonography During Early-Stage Ovarian Cancer in Laying Hens A Preclinical Model of Human Spontaneous Ovarian Cancer

Objective. Tumor‐associated neoangiogenesis (TAN) is one of the earliest events in ovarian tumor growth and represents a potential target for early detection of ovarian cancer (OVCA). Because it is difficult to identify patients with early‐stage OVCA, the goal of this study was to explore a spontaneous animal model of in vivo ovarian TAN associated with early‐stage OVCA detectable by Doppler ultrasonography (DUS). Methods. White Leghorn laying hens were scanned transvaginally at 15‐week intervals up to 45 weeks. Gray scale ovarian morphologic characteristics and Doppler indices were recorded. Hens were euthanized at diagnosis for ultrasonographic morphologic/vascular abnormalities or at the end of the study (those that remained normal). Ovarian morphologic and histologic characteristics were evaluated. Vascular endothelial growth factor (VEGF) and αvβ3‐integrin expression was assessed by immunohistochemical analysis. Doppler ultrasonographic observations were compared with histologic and immunohisto‐chemical findings to determine the ability of DUS to detect ovarian TAN. Results. Significant changes in ovarian blood flow parameters were observed during transformation from normal to tumor development in the ovary (P < .05). Tumor‐related changes in ovarian vascularity were identified by DUS before the tumor became detectable by gray scale imaging. Increased expression of VEGF and αvβ3‐integrins was associated with tumor development. Ovarian TAN preceded tumor progression in hens. Conclusions. The results suggest that ovarian TAN may be an effective target for the detection of early‐stage OVCA. The laying hen may also be useful for studying the detection and inhibition of ovarian TAN using various means, including the efficacy of contrast agents, targeted molecular imaging, and antiangiogenic therapies.