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Black women in the United States are disproportionately affected by early-onset, triple-negative breast cancer. DNA methylation has shown differences by race in healthy and tumor breast tissues. We examined associations between genome-wide DNA methylation levels in breast milk and breast cancer risk factors, including race, to explain how this reproductive stage influences a woman's risk for, and potentially contributes to racial disparities in, breast cancer. Breast milk samples and demographic, behavioral, and reproductive data, were obtained from cancer-free, uniparous, and lactating U.S. black (n = 57) and white (n = 82) women, ages 19–44. Genome-wide DNA methylation analysis was performed on extracted breast milk DNA using the Infinium HumanMethylation450 BeadChip. Statistically significant associations between breast cancer risk factors and DNA methylation beta values, adjusting for potential confounders, were determined using linear regression followed by Bonferroni Correction (P < 1.63 × 10−7). Epigenetic analysis in breast milk revealed statistically significant associations with race and lactation duration. Of the 284 CpG sites associated with race, 242 were hypermethylated in black women. All 227 CpG sites associated with lactation duration were hypomethylated in women who lactated longer. Ingenuity Pathway Analysis of differentially methylated promoter region CpGs by race and lactation duration revealed enrichment for networks implicated in carcinogenesis. Associations between DNA methylation and lactation duration may offer insight on its role in lowering breast cancer risk. Epigenetic associations with race may mediate social, behavioral, or other factors related to breast cancer and may provide insight into potential mechanisms underlying racial disparities in breast cancer incidence.

Introduction: Identifying biomarkers of breast cancer risk among young women would have value in developing effective screening and prevention strategies at early ages. We have proposed that DNA methylation analysis of breast milk may provide breast cancer risk information among young women, and could possibly provide etiologic clues related to the higher rates of early onset cancers among African American as compared with White women in the US. Objective: The purpose of this project was to identify associations between genome-wide DNA methylation levels in breast milk and race adjusted for other breast cancer risk factors. Study Population: Cancer-free, lactating U.S. Black (n=57) and White (n=82) women, ages 19 to 44, provided frozen breast milk samples, as well as demographic, behavioral, and reproductive data, to the Breastmilk Laboratory at University of Massachusetts Amherst. Women were uniparous and did not have a personal history of breast cancer at the time of milk donation. Methods: DNA was extracted from breast milk samples using the phenol-chloroform method. Genome-wide methylation analysis was performed on breast milk samples using the Infinium HumanMethylation450 BeadChip. Probes with 50% or more missing data, cross-reactive probes, as well as probes with minor allelic frequency greater than 0.05 in European- or African-Americans were removed, leaving 379,042 CpG sites for analysis. Multivariate generalized linear regression models were used to examine associations between race and other breast cancer risk factors and methylation beta values, adjusting for potential confounding factors. P-values less than 1E-7 were considered statistically significant. Results: Black women in this study were more likely to be never smokers, to not have used over-the-counter pain medication in the past week, and to breastfeed longer. After adjustment by age, BMI, smoking status, and batch number, race was significantly associated with differential methylation at 1143 CpG sites, including 1024 at which Black women demonstrated increased methylation levels. Additionally, breastfeeding duration was associated with 269 CpG sites, with 268 showing a significant inverse relationship with methylation. Methylation sites significantly associated with Black race and lactation duration were located within tumor suppressor and promoter genes as well as in genes implicated in obesity and diabetes. Conclusion: This preliminarily analysis of DNA methylation in breast milk suggests that Black women have increased methylation and longer breastfeeding is associated with reduced methylation. Further research to understand how etiologic factors related to breast cancer may alter DNA methylation patterns in normal breast may lead to improved understanding of breast cancer risk at a young age and potentially causes of racial disparities in breast cancer incidence between White and Black women. Citation Format: Davis Lynn BC, Bodelon C, Pfeiffer RM, Yang HP, Yang H, Lee M, Laird PW, Campan M, Weisenberger DJ, Murphy J, Sherman ME, Browne EP, Anderton DL, Arcaro KF, Gierach GL. Differences in genome-wide DNA methylation levels in breast milk by race and lactation duration [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr P6-10-07.

BackgroundPrior candidate gene studies have shown tumor suppressor DNA methylation in breast milk related with history of breast biopsy, an established risk factor for breast cancer. To further establish the utility of breast milk as a tissue-specific biospecimen for investigations of breast carcinogenesis we measured genome-wide DNA methylation in breast milk from women with and without a diagnosis of breast cancer in two independent cohorts.MethodsDNA methylation was assessed using Illumina HumanMethylation450k in 87 breast milk samples. After quality control, 368,171 autosomal CpG loci were analyzed. Cell type proportion estimates from RefFreeCellMix were calculated and adjusted for in this Epigenome Wide Association Study using linear mixed effects models adjusted for history of breast biopsy, age, time of delivery, cell type proportion estimates, array chip, and subject as random effect.ResultsEpigenome-wide analyses identified 58 differentially methylated CpG sites associated with a breast cancer diagnosis in the prospectively collected milk samples from the breast that would develop cancer compared with women without a diagnosis of breast cancer (q-value < 0.05). Nearly all CpG sites associated with a breast cancer diagnosis were hypomethylated in cases compared with controls, and were enriched for CpG islands. In addition, inferred repeat element methylation was lower in breast milk DNA from cases compared to controls, and cases exhibited increased estimated epigenetic mitotic tick rate as well as DNA methylation age compared with controls.ConclusionBreast milk has utility as a biospecimen for prospective assessment of disease risk, for understanding the underlying molecular basis of breast cancer risk factors, and improving primary and secondary prevention of breast cancer.

BACKGROUND: Analysis of cytokines and growth factors in human milk offers a noninvasive approach for studying the microenvironment of the postpartum breast, which may better reflect tissue levels than testing blood samples. Given that Black women have a higher incidence of early onset breast cancers than White women, we hypothesized that milk of the former contains higher levels of pro-inflammatory cytokines, adipokines, and growth factors. METHODS: Participants included 130 Black and 162 White women without a history of a breast biopsy who completed a health assessment questionnaire and donated milk for research. Concentrations of 15 analytes in milk were examined using two multiplex and 4 single-analyte electrochemiluminescent sandwich assays to measure pro-inflammatory cytokines, angiogenesis factors, and adipokines. Mixed-effects ordinal logistic regression was used to identify determinants of analyte levels and to compare results by race, with adjustment for confounders. Factor analysis was used to examine covariation among analytes. RESULTS: Thirteen of 15 analytes were detected in ≥25% of the human milk specimens. In multivariable models, elevated BMI was significantly associated with increased concentrations of 5 cytokines: IL-1β, bFGF, FASL, EGF, and leptin (all p-trend < 0.05). Black women had significantly higher levels of leptin and IL-1β, controlling for BMI. Factor analysis of analyte levels identified two factors related to inflammation and growth factor pathways CONCLUSION: This exploratory study demonstrated the feasibility of measuring pro-inflammatory cytokines, adipokines and angiogenesis factors in human milk, and revealed higher levels of some pro-inflammatory factors, as well as increased leptin levels, among Black as compared with White women.

TGF-β plays a dual role in breast carcinogenesis, acting at early stages as tumor-suppressors and later as tumor-promoters. TGF-β isoforms are expressed in breast tissues and secreted in milk, suggesting that analysis of levels in milk might be informative for breast cancer risk. Accordingly, we assessed TGF-β2 levels in milk from women who had undergone a breast biopsy and related the concentrations to diagnosis. Milk donated by women who had undergone or were scheduled for a breast biopsy was shipped on ice for processing and testing. Breast cancer risk factors were obtained through a self-administered questionnaire, and biopsy diagnoses were extracted from pathology reports. TGF-β2 levels in milk, assessed as absolute levels and in relation to total protein, were analyzed in bilateral samples donated by 182 women. Linear regression was used to estimate relationships of log-transformed TGF-β2 levels and TGF-β2/ total protein ratios to biopsy category. Milk TGF-β2 levels from biopsied and non-biopsied breasts within women were highly correlated (r 2 = 0.77). Higher mean TGF-β2 milk levels (based on average of bilateral samples) were marginally associated with more severe breast pathological diagnosis, after adjusting for duration of nursing current child (adjusted p trend = 0.07). Our exploratory analysis suggests a borderline significant association between higher mean TGF-β2 levels in breast milk and more severe pathologic diagnoses. Further analysis of TGF-β signaling in milk may increase understanding of postpartum remodeling and advance efforts to analyze milk as a means of assessing risk of breast pathology.

Background Diets rich in fruits and vegetables (F/V) can reduce the inflammatory profile of circulating cytokines and potentially decrease the risk of breast cancer. However, the extent to which a diet rich in F/V alters cytokine levels in breast tissue remains largely unknown. Breast milk provides a means of assessing concentrations of secreted cytokines in the breast microenvironment and is a potential tool for studying the effects of diet on inflammation in breast tissue and breast cancer risk. Objective The aim of this pilot randomized trial was to test the feasibility of increasing F/V intake in breastfeeding women and of measuring changes in markers of inflammation in breast milk. Design and intervention Participants randomized to the intervention (n=5) were provided weekly boxes of F/V, along with dietary counseling, to increase consumption of F/V to 8 to 10 daily servings for 12 consecutive weeks. Controls (n=5) were directed to the US Department of Agriculture’s “ChooseMyPlate” diet for pregnancy and breastfeeding. Participants/setting Ten breastfeeding women consuming fewer than five servings of F/V per day, as estimated by the National Institutes of Health “All-Day” Fruit and Vegetable Screener (F/V Screener), were recruited through flyers and a lactation consultant between February and May 2016 in the Western Massachusetts area. Main outcome measures Baseline demographic and F/V intake data were collected during enrollment. At week 1 and week 13 (final) home visits, participants provided milk samples and anthropometric measurements were recorded. Participants completed F/V screeners at baseline and at study end. Adiponectin, leptin, C-reactive protein, and 11 additional cytokines were measured in breast milk collected at weeks 1 and 13. Statistical analyses F/V consumption at baseline and after the final visit, and between controls and intervention groups, was compared with dependent and independent t tests, respectively. Differences between cytokine levels at weeks 1 and 13 were assessed with a mixed-effects repeated-measures model. Results All women in the intervention increased F/V intake and were consuming more servings than controls by week 13; daily serving of F/V at baseline and final visit: controls=1.6 and 2.0, diet=2.6 and 9.9. Most cytokines were detected in the majority of milk samples: 12 were detected in 90% to 100% of samples, one was detected in 75% of samples, and one was detected in 7.5% of samples; coefficients of variation were below 14% for 11 of the cytokines. Conclusions These preliminary findings indicate that it is feasible to significantly increase F/V intake in breastfeeding women and provide support for conducting a larger diet intervention study in breastfeeding women, in which longer-term benefits of the intervention are assessed.

S-phase kinase-associated protein 2 (SKP2) is an important cell cycle regulator, targeting the cyclin-dependent kinase (CDK) inhibitor p27 for degradation, and is frequently overexpressed in breast cancer. p27 regulates G1 /S transition by abrogating the activity of cyclin/CDK complexes. p27 can undergo phosphorylation at serine 10 (pSer10p27). This phosphorylation event is associated with increased cell proliferation and poor prognosis in patients with glioma. The relationship between SKP2 and pSer10p27 in breast cancer has not been previously investigated. Immunohistochemistry (IHC) of SKP2, p27, pSer10p27, and other genes involved in this pathway, was analyzed in 188 breast tumors and 50 benign reduction mammoplasty samples. IHC showed SKP2 to be more highly expressed in estrogen receptor α (ERα)-negative breast cancers and demonstrated that triple-negative tumors were more likely to have high expression of SKP2 than were non-triple negative, ERα-negative tumors. A significant positive relationship was discovered for SKP2 and pSer10p27. High levels of SKP2 and pSer10p27 were observed significantly more often in ERα-negative and triple-negative than in ERα-positive breast cancers. Use of the triple-negative TMX2-28 breast cancer cell line to address the role of SKP2 in cell cycle progression confirmed that SKP2 contributes to a more rapid cell cycle progression and may regulates pSer10p27 levels. Together, the results indicate that presence of high SKP2 plus high pSer10p27 levels in triple-negative breast cancers is associated with aggressive growth, and highlight the validity of using SKP2 inhibitors as a therapeutic approach for treating this subset of breast cancers.

Roughly two-thirds of all breast cancers are ERα-positive and can be treated with the antiestrogen, Tamoxifen, however resistance occurs in 33% of women who take the drug for more than 5 y. Aberrant DNA methylation, an epigenetic mechanism that alters gene expression in cancer, is thought to play a role in this resistance. To develop an understanding of Tamoxifen-resistance and identify novel pathways and targets of aberrant methylation, DNA from MCF-7 breast cancer cells and Tamoxifen-resistant derivatives, TMX2-11 and TMX2-28, were analyzed using the Illumina HumanMethylation450 BeadChip. Normalizing against MCF-7 values, ERα-positive TMX2-11 had 4000 hypermethylated sites and ERα-negative TMX2-28 had over 33 000. Analysis of CpG sites altered in both TMX2-11 and TMX2-28 revealed that the Tamoxifen-resistant cell lines share 3000 hypermethylated and 200 hypomethylated CpGs. ZNF350 and MAGED1, two genes hypermethylated in both cell lines, were examined in greater detail. Treatment with 5-aza-2ꞌdeoxycitidine caused a significant reduction in promoter methylation of both ZNF350 and MAGED1 and a corresponding increase in expression in TMX2-28. A similar relationship between methylation and expression was not detected in TMX2-11. Our findings are indicative of the variable responses to methylation-targeted breast cancer therapy and highlight the need for biomarkers that accurately predict treatment outcome.

Background: Breast cancer risk increases during pregnancy and remains elevated for a number of years thereafter. Cancer-associated proteins that are secreted into breast milk may provide a means to detect cancer in the lactating breast or to assess future breast cancer risk. Objective: To determine whether proteins linked to breast cancer would be differentially expressed in matched (both breasts from each participant) human milk samples collected from women with unilateral breast cancer. Methods: Five cancer-associated proteins (basic fibroblast growth factor [bFGF], YKL-40, neutrophil gelatinase-associated lipocalin, and transforming growth factor β1 and β2) were analyzed in milk provided by 5 lactating women, 4 of whom were known to have cancer in 1 breast (and the opposite breast clinically disease free) at the time of milk collection and 1 who developed breast cancer 2 years after milk collection. Results: Expression was significantly higher for TGFβ2 ( P = .03) and bFGF ( P =.03) in the breasts with cancer. Conclusion: These proteins may play a role in assessing a woman’s risk of pregnancy-associated breast cancer. Because of variable protein concentration among patients and the limited sample size, the results are considered preliminary.

Accurately identifying women at increased risk of developing breast cancer will provide greater opportunity for early detection and prevention. DNA promoter methylation is a promising biomarker for assessing breast cancer risk. Breast milk contains large numbers of exfoliated epithelial cells that are ideal for methylation analyses. Exfoliated epithelial cells were isolated from the milk obtained from each breast of 134 women with a history of a non-proliferative benign breast biopsy (Biopsy Group). Promoter methylation of three tumor suppressor genes, RASSF1, SFRP1 and GSTP1, was assessed by pyrosequencing of bisulfite-modified DNA. Methylation scores from the milk of the 134 women in the Biopsy Group were compared to scores from 102 women for whom a breast biopsy was not a recruitment requirement (Reference Group). Mean methylation scores for RASSF1 and GSTP1 were significantly higher in the Biopsy than in the Reference Group. For all three genes the percentage of outlier scores was greater in the Biopsy than in the Reference Group but reached statistical significance only for GSTP1. A comparison between the biopsied and non-biopsied breasts of the Biopsy Group revealed higher mean methylation and a greater number of outlier scores in the biopsied breast for both SFRP1 and RASSF1, but not for GSTP1. This is the first evidence of CpG island methylation in tumor suppressor genes of women who may be at increased risk of developing breast cancer based on having had a prior breast biopsy.

Prior candidate gene studies have shown that methylation-induced silencing of tumor suppressor genes in breast milk is related with history of breast biopsy, an established risk factor for breast cancer. To further establish the utility of breast milk as a tissue-specific biospecimen for investigations of breast carcinogenesis we measured genome-wide DNA methylation in breast milk from women with and without a diagnosis of breast cancer. Breast milk provides a source of cells without invasive procedure that is available during a window of increased breast cancer risk. Actively lactating women were enrolled and consented through the University of Massachusetts Amherst breast milk study targeting high risk subjects. Participants completed a questionnaire to provide demographic and breast cancer risk factor data. Genome-wide DNA methylation levels were quantified using the Illumina 450K HumanMethylation array. Data was processed in R using RnBeads. Beta-values were normalized (FUNnorm), low quality probes were filtered and transformed to M-values. Linear mixed effects models were fit including a random effect for subject and adjusted for history of breast biopsy, age, body mass index, age of the baby, and estimated cell type proportions, an interaction term for time of diagnosis was included. DNA methylation in women with breast cancer (n=9), was compared with women without a history of breast cancer (n=22). Among women with a diagnosis of breast cancer there were two groups: those diagnosed within one year of donating breast milk, and those diagnosed more than one year before sample collection. Four women in the control and five women in the cancer group provided bilateral breast milk samples at recruitment and both samples were included in the analyses. CpGs with significant differential methylation were tested for enrichment in gene pathways using the KEGG database. We identified 215 differentially methylated CpG sites in women diagnosed within one year of sample donation compared with women without a diagnosis of breast cancer (Q Citation Format: Lucas A. Salas, Sara N. Lundgren, Eva P. Browne, Elizabeth C. Punska, Douglas L. Anderton, Kathleen F. Arcaro, Brock C. Christensen. Altered DNA methylation in breast milk from women with breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4355. doi:10.1158/1538-7445.AM2017-4355

Background: Breast cancer is the most frequently diagnosed cancer among Turkish women and both the incidence and associated mortality appear to be increasing. Of particular concern is the percentage of young women diagnosed with breast cancer; roughly 20% of all breast cancer diagnoses in Turkey are in women younger than 40 years. Increased DNA methylation in the promoter region of tumor suppressor genes is a promising molecular biomarker, and human milk provides exfoliated breast epithelial cells appropriate for DNA methylation analyses. Comparisons between DNA methylation patterns in epithelial (epithelial-enriched) and nonepithelial (epithelial-depleted) cell fractions from breast milk have not been reported previously. Objective: In the present study, we examined promoter methylation of 3 tumor suppressor genes in epithelial-enriched and epithelial-depleted cell fractions isolated from breast milk of 43 Turkish women. Methods: Percentage methylation in the promoter region of Rass association domain family 1 (RASSF1), secreted frizzle related protein 1 (SFRP1), and glutathione- S-transferase class pi 1 was determined by pyrosequencing of the epithelial-enriched and epithelial-depleted cell fractions. Results: Pyrosequencing identified a few subjects with significantly increased methylation in 1 or more genes. There was little correlation between the 2 cell fractions within individuals; only 1 woman had increased methylation for 1 gene (SFRP1) in both her enriched and depleted cell fractions. Methylation was positively associated with age for SFRP1 (epithelial-depleted fraction) and with body mass index for RASSF1 (epithelial-enriched cell fraction), respectively. Conclusion: Overall, results show that the methylation signals vary between different cell types in breast milk and suggest that breast milk can be used to assess DNA methylation patterns associated with increased breast cancer risk.

Background Paralemmin-1 is a phosphoprotein lipid-anchored to the cytoplasmic face of membranes where it functions in membrane dynamics, maintenance of cell shape, and process formation. Expression of paralemmin-1 and its major splice variant (Δ exon 8) as well as the extent of posttranslational modifications are tissue- and development-specific. Paralemmin-1 expression in normal breast and breast cancer tissue has not been described previously. Results Paralemmin-1 mRNA and protein expression was evaluated in ten breast cell lines, 26 primary tumors, and 10 reduction mammoplasty (RM) tissues using real time RT-PCR. Paralemmin-1 splice variants were assessed in tumor and RM tissues using a series of primers and RT-PCR. Paralemmin-1 protein expression was examined in cell lines using Western Blots and in 31 ductal carcinomas in situ, 65 infiltrating ductal carcinomas, and 40 RM tissues using immunohistochemistry. Paralemmin-1 mRNA levels were higher in breast cancers than in RM tissue and estrogen receptor (ER)-positive tumors had higher transcript levels than ER-negative tumors. The Δ exon 8 splice variant was detected more frequently in tumor than in RM tissues. Protein expression was consistent with mRNA results showing higher paralemmin-1 expression in ER-positive tumors. Conclusions The differential expression of paralemmin-1 in a subset of breast cancers suggests the existence of variation in membrane dynamics that may be exploited to improve diagnosis or provide a therapeutic target.

Promoter methylation analysis of genes frequently silenced in breast cancer is a promising indicator of breast cancer risk, as these methylation events are thought to occur long before presentation of disease. The numerous exfoliated epithelial cells present in breast milk may provide the breast epithelial DNA needed for detailed methylation analysis and assessment of breast cancer risk. Fresh breast milk samples and health, lifestyle, and reproductive history questionnaires were collected from 111 women. Pyrosequencing analysis was conducted on DNA isolated from the exfoliated epithelial cells immunomagnetically separated from the total cell population in the breast milk of 102 women. A total of 65 CpG sites were examined in six tumor suppressor genes: PYCARD (also known as ASC or TMS1), CDH1, GSTP1, RBP1 (also known as CRBP1), SFRP1, and RASSF1. A sufficient quantity of DNA was obtained for meaningful analysis of promoter methylation; women donated an average of 86 ml of milk with a mean yield of 32,700 epithelial cells per ml. Methylation scores were in general low as expected of benign tissue, but analysis of outlier methylation scores revealed a significant relationship between breast cancer risk, as indicated by previous biopsy, and methylation score for several CpG sites in CDH1, GSTP1, SFRP1, and RBP1. Methylation of RASSF1 was positively correlated with women's age irrespective of her reproductive history. Promoter methylation patterns in DNA from breast milk epithelial cells can likely be used to assess breast cancer risk. Additional studies of women at high breast cancer risk are warranted.

Nearly all developed countries have experienced dramatic declines in the seasonality of demographic behavior over the last century. This article examines seasonal behavior in the population of Taiwan between 1906 and 1942, during Japanese colonial rule. First, we analyze seasonal demographic cycles and their ties to the cultural calendar and agriculture. Second, we compare seasonality in two regions (Xinzhu and Tainan) that experienced different agricultural development under the colonial regime. Findings demonstrate that demographic seasonality was less pronounced when urban development and colonial agricultural intensification produced more heterogeneous seasonal patterns of labor. We find that changes in agricultural organization, rather than purely a shift to nonagricultural production, may significantly reduce established seasonal timing of demographic behavior.

Data on the prevalence, magnitude and causes of blindness and severe visual impairment in children are needed for planning and evaluating preventive and curative services for children, and for planning special education and low vision services. Prevalence data can be obtained from a variety of different sources, each of which has limitations. The available data suggest that there may be a ten-fold difference in prevalence between the wealthiest countries of the world and the poorest, ranging from as low as 0.1/1000 children aged 0-15 years in the wealthiest countries to 1.1/1000 children in the poorest. In this paper, the available data are reviewed and the epidemiological methods and findings are discussed.

Background: TGF-β isoforms play a dual role in breast carcinogenesis, acting at early stages as tumor-suppressors and later as tumor-promoters. TGF-β isoforms are expressed in breast tissues and secreted in milk, suggesting that analysis of levels in milk might have value for understanding the effect of pregnancy and lactation on breast carcinogenesis. Data suggest that TGF-β2 levels in milk are higher than TGF-β1, show heterogeneity among women, and may vary with psychosocial stress, maternal immunity, and length of nursing. We assessed whether TGF-β2 levels in milk from each breast were correlated with a history of prior breast biopsy (biopsied vs. not biopsied) and biopsy diagnosis. Methods: Fresh breast milk donated by lactating women who had either undergone or were scheduled for a breast biopsy was shipped on ice to a central laboratory. Breast cancer risk factors were obtained via questionnaire and biopsy diagnoses were extracted from pathology reports. Milk was processed to create cellular-enriched and cellular-depleted fractions and frozen at -20[≤]C prior to testing. TGF-β2 levels in milk were analyzed using the Quantikine Human TGF-β2 Immunoassay (R&D Systems DB250) and quantified using the Coomassie Plus (Bradford) Assay (Thermo Scientific) kits in bilateral samples donated by182 women, including duplicate samples from 1 woman (n= 366). We compared log transformed TGF-β2 (pg/ml) and TGF-β2/total protein ratio (pg/μg) levels for biopsied and non-biopsied breasts using t-tests and ordinary least squares regressions adjusted for confounders and clustering of observations by woman. Results: TGF-β2 levels ranged 0 - 68,070 pg/ml and total protein levels ranged 1,558- 36,198 μg/ml. The levels of TGF-β2 in the biopsied and non-biopsied breasts were correlated (r2 = 0.64), albeit not statistically significant. Controlling for mother's attained age, age at first birth, age of baby (surrogate for length of nursing), number of live births, time since breast biopsy, and race, did not alter the finding for TGF-β2 or the TGF-β2/total protein ratio. But, mean levels of TGF-β2 proteins were highest in milk among women with cancer (9971 pg/ml, n=16), followed by proliferative lesions with atypia (8842 pg/ml; n=4) biopsy diagnoses. TGF-β2 levels were lower among women with non-proliferative biopsy diagnoses (4131 pg/ml; n=292) as compared with those who had proliferative lesions with atypia or cancer (P Conclusion: More severe pathologic diagnoses were associated with higher mean TGF-β2 levels in breast milk, although levels of biopsied and non-biopsied breasts were correlated. These data suggest that higher TGF-β2 levels in milk are linked to higher breast cancer risk for a woman, but do not specifically indicate which breast is at risk. Citation Format: Hannah P. Yang, Mark E. Sherman, Sallie Smith Schneider, Christina M. Chisholm, Eva P. Browne, Sidra Mahmood, Sarah Lenington, Douglas L. Anderton, Kathleen A. Arcaro. Association of TGF-β2 levels in breast milk with breast biopsy diagnosis. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 117. doi:10.1158/1538-7445.AM2013-117

Estrogen receptor (ER) status remains one of the most important breast cancer diagnostic and prognostic biomarkers. Roughly two-thirds of all breast cancers are ER-positive and can be treated with the anti-estrogen, Tamoxifen, however resistance occurs in 33% of women who take the drug for more than 5 years. MCF-7 breast cancer cells were previously cultured with 1 uM Tamoxifen, and after 6 months the clonal lines TMX2-11 and TMX2-28 with acquired resistance were isolated. TMX2-28 lost ER-α gene expression while a second clonal line, TMX2-11, did not. TMX2-28 cells are highly invasive in vitro, in contrast to TMX2-11 and the parent cell line, MCF-7. Studies of endocrine resistance and global gene expression in Tamoxifen-resistant cell cultures and human tumors have detected alterations in numerous pathways including ER-signaling, growth factor receptor signaling, cell cycle, and apoptosis. Aberrant DNA promoter methylation is a major epigenetic mechanism affecting gene expression in cancer. Our focus is on its role in Tamoxifen resistance. Hypermethylation of normally unmethylated cytosines in CpG-rich promoter regions frequently results in gene silencing, while hypomethylation in CpGs outside promoter regions leads to genetic instability. To develop a broad understanding of Tamoxifen resistance and to identify novel pathways and targets of aberrant methylation, DNA from MCF-7, TMX2-11, and TMX2-28 was analyzed using the Illumina HumanMethylation450 BeadChip. The three cell lines have similar hypermethylation patterns in 306,000 CpG sites and similar hypomethylation patterns in 132,000 CpG sites across the genome. A subset of CpG sites was identified where the two Tamoxifen-selected lines share a methylation pattern distinct from the MCF-7 parent line. When normalized to MCF-7, TMX2-11 and TMX2-28 share 722 hypermethylated and 65 hypomethylated CpG sites in the promoter region encompassing 539 and 55 genes respectively. Pathway analysis indicates the hypermethylated genes are involved in processes relevant to acquired Tamoxifen resistance. These pathways include cell signaling, adhesion, transcriptional activation and repression, differentiation, proliferation, and apoptosis. Ongoing studies on a subset of the genes found differentially methylated between parental and Tamoxifen-selected cell lines are aimed at confirming methylation findings and examining gene expression. Additionally, the cells will be treated with the demethylating agent 5-aza-deoxycitidine to determine whether reversal of the hypermethylation restores Tamoxifen sensitivity. Greater knowledge of the molecular modifications accompanying Tamoxifen-resistance is needed, as it will lead to discovery of new therapeutic targets and improved treatment, possibly allowing prediction of resistance from biopsy DNA. Citation Format: Kristin E. Williams, Douglas L. Anderton, Maxwell P. Lee, Brian T. Pentecost, Kathleen F. Arcaro. Epigenetic changes in breast cancer cells associated with acquired Tamoxifen resistance. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5368. doi:10.1158/1538-7445.AM2013-5368

Breast cancer is expected to be the most frequently diagnosed form of cancer and the second leading cause of cancer deaths among women in 2012. The primary objective of the present study was to provide insights into the molecular mechanisms underlying the aggressive growth of triple-negative and basal-like breast cancers. In this study, we use the Tamoxifen-selected, MCF-7 derivative, TMX2-28 breast cancer cell line. TMX2-28 cells are triple-negative, have acquired a mixed basal/luminal cytokeratin profile, suggestive of a more basal-like phenotype, and are highly proliferative. We also examined thirty frozen breast carcinoma samples for mRNA, and 188 breast tumors and 50 benign reduction mammoplasty samples (formalin-fixed paraffin embedded) for protein expression. S-phase kinase-associated protein 2 (SKP2) plays an important role in cell cycle regulation by targeting p27 for degradation. The cyclin-dependent kinase (CDK) inhibitor p27 regulates G1/S transition by binding cyclin/CDK complexes and abrogating its activity. By targeting p27 for degradation, SKP2 frees the complexes needed to progress into the S phase of the cell cycle. Evaluation of SKP2 expression in TMX2-28 revealed significantly higher levels than in other breast cancer cell lines. Despite the high levels of SKP2 expression, p27 protein was not reduced. However, levels of the serine-10 phosphorylated form of p27 (pSer10p27), which has been associated with increased proliferation rates, was found to be upregulated. Furthermore, suppression of SKP2 completely eliminated pSer10p27 expression and slowed cell cycle progression confirming the role of SKP2 in the aggressive growth of TMX2-28 cells. Assessment of mRNA from 30 frozen human breast cancers demonstrated that SKP2 is more highly expressed in ERα-negative and basal-like breast cancers. Immunohistochemical analysis confirmed that SKP2 is more highly expressed in ERα-negative breast cancers and for the first time demonstrated that triple-negative breast cancers are more likely to overexpress SKP2 than are non-triple negative, but still ERα-negative tumors. We did not observe an inverse relationship between SKP2 and p27 expression, however a significant positive relationship was discovered between SKP2 and pSer10p27. Furthermore, high levels of SKP2 and pSer10p27 were observed significantly more often in ERα-negative and triple-negative than in ERα-positive breast cancers. The relationship between SKP2 and pSer10p27 is a better indicator of increased tumor proliferation in triple-negative breast cancers based on these results and those of our cell culture model. Inhibiting SKP2 in tumors expressing high levels of both SKP2 and pSer10p27 may be a valid therapeutic approach. Citation Format: Katerina D. Fagan-Solis, Joseph M. Gogzit, Brook A. Bentley, Sharon M. Marconi, Christopher N. Otis, Douglas L. Anderton, Sallie Smith Schneider, Brian T. Pentecost, Kathleen F. Arcaro. SKP2 overexpression is associated with increased serine-10 phosphorylation of p27 (pSer10p27) in triple-negative breast cancers. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 563. doi:10.1158/1538-7445.AM2013-563

Breast cancer is the second most commonly diagnosed cancer among women, and is difficult to diagnose during pregnancy and lactation. Frequent methylation of the promoter region of key tumor suppressor genes occurs in the early etiology of breast cancer. We aimed to evaluate the feasibility of using DNA extracted from the epithelial cells exfoliated in breast milk as a biomarker for breast cancer risk. To date, studies of methylation patterns in breast epithelial cells from asymptomatic women have been limited to cells obtained from fine nipple aspirate and ductal lavage. Analysis of promoter hypermethylation in exfoliated epithelial cells isolated from breast milk provides a non-invasive opportunity to assess breast cancer risk. Breast milk samples from both the biopsied and non-biopsied breasts of women scheduled for or who had had a breast biopsy were collected and processed within 24 hours of expression. Epithelial enriched cell fractions were isolated via immunomagnetic separation and DNA subsequently isolated and bisulfite treated for RASSF1, GSTP1, and SFRP1 promoter methylation analysis via pyrosequencing. When available, pathology reports from each participant were also obtained to categorize the outcome of the biopsy. Over the past two years, we have collected breast milk samples from 230 women who were scheduled for or had a breast biopsy. Of those 230 participants, we obtained the biopsy reports from 85% of participants (196 biopsy reports). Using pyrosequencing, we analyzed 30 individual CpG sites on 3 genes (RASSF1, GSTP1 and SFRP1) in epithelial enriched cell fractions of the biopsied and non-biopsied breast. In women whose biopsy resulted in a non-proliferative lesion (n=104), there was no significant difference in the average epithelial DNA methylation of their biopsied breast compared to their non-biopsied breast for RASSF1 (p=0.15) and GSTP1 (p=0.92), but for SFRP1 the average methylation was significantly higher in the biopsied breast (p = 0.02). In women whose biopsy result revealed cancer (n=10), there was a significant increase in average RASSF1 methylation of their biopsied breast compared to their non-biopsied breast (p=0.05). This study is the first to collect breast milk and obtain epithelial cells and DNA from a population of women at increased risk of breast cancer. Utilizing this unique population, we have been able to assess promoter methylation in 3 genes known to be methylated in breast cancer. Long term follow-up is ongoing for women enrolled in this study to determine breast health outcomes. Knowledge of promoter methylation along with long term follow-up may help us to better assess individual breast cancer risk. Given the high percentage of women that give birth and therefore produce milk, if even for just a few days, a breast cancer risk screen based on epithelial cells in milk has the potential to affect a significant portion of all women. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-443. doi:10.1158/1538-7445.AM2011-LB-443

Breast cancers discovered during pregnancy or lactation (gestational) are known to be very aggressive and of poor prognosis. We have found a Mouse Mammary Tumor Virus (MMTV) like virus in human breast cancer, but not in adjacent normal tissue, indicating it exogenous origin. This virus, Human Mammary Tumor Virus (HMTV) recognized by its unique env sequence is present in 62% of gestational breast cancer, compared with 38% of sporadic breast cancers (Wang et al, 2003. Medical Oncology 20:233-236). The aggressive phenotype of gestational breast cancer was suggested to be due to the effect of hormones that stimulate cellular growth. HMTV- LTR contained sequences which are responsive to several hormones (glucocorticoid, estrogen, prolactin, and progesterone) and a transcription factor (TEF-1 family) (Wang et al, 2003. Medical Oncology 20:233-236). To find if HMTV is present in other tissues that respond to hormones, we have investigated the presence of HMTV sequences in two of these tissues. Endometrium is a classical example of such tissue. Endometrial cancers and normal endometrium were analyzed for HMTV env sequences. The results indicated that HMTV env sequences were present in 3.4% of the normal tissues and 37% of the tumors analyzed. Recently, it was reported that MMTV-like sequence was found in 10% of endometrial cancers (Johal et al. 2010, J. Med. Virology; 82: 1044-50). During lactation, the mammary gland experiences major metabolic changes due to hormonal stimulation. We have examined 88 normal milks and 63 milks from women, who have undergone biopsies. Whereas in the cells of normal milks 8% showed env sequences cells in the milk of the biopsied group showed that 23.8% contain HMTV sequences. These results confirmed electron microscope observations that a betaretrovirus was present in milk from healthy women and those with breast cancer (Moore et al. 1971, Nature 229: 611-615). Taken together, the results presented here suggest that tissues and tumors stimulated by hormones showed high prevalence of HMTV sequences. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2302. doi:10.1158/1538-7445.AM2011-2302

Gene specific promoter methylation in breast epithelial cells is considered one of the most promising biomarkers for assessing breast cancer risk. Unfortunately, obtaining breast epithelial cells is difficult. We recently demonstrated that human breast milk provides a source of epithelial cells useful for assessing breast cancer risk through analysis of DNA promoter methylation patterns. However, it has not been determined whether separation of the epithelial cells from the total cell population is necessary to obtain reliable promoter methylation patterns useful in assessing this risk. The present study determines the extent to which separation of the cell populations in breast milk alters the observed methylation patterns. A fresh breast milk sample (average size of 118 mL ± 29 mL) was collected from six lactating women between 25 and 43 years of age. For each milk sample, three groups of cells were collected: an unseparated (or total) cell population, an epithelial-enriched cell fraction and an epithelial-depleted cell fraction. The depleted and enriched cell fractions were obtained by immunomagnetic separation. Cell fractions were frozen and shipped to EpigenDX for DNA isolation, bisulfite treatment and pyrosequencing. Promoter methylation of five tumor suppressor genes (APC, CyclinD2, p16, RARβ, and RASSF1A) was examined. Cell separation resulted in distinct methylation patterns. Overall, the highest methylation scores were detected in the epithelial-enriched cell fractions: mean methylation scores were significantly higher in the epithelial-enriched fraction as compared to the depleted fraction or total cell population for APC, CyclinD2, RARβ, and RASSF1A. Only p16 showed very low methylation: Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1995. doi:10.1158/1538-7445.AM2011-1995

Analysis of promoter methylation patterns in genes frequently downregulated in breast cancer may serve as an early indicator of breast cancer risk. To date, studies of methylation patterns in breast epithelial cells from asymptomatic women have been limited to cells obtained from fine nipple aspirate and ductal lavage. Analysis of promoter hypermethylation in exfoliated epithelial cells isolated from breast milk provides a non-invasive opportunity to assess breast cancer risk. Our preliminary data indicate that we can detect a cancer methylation phenotype in the milk of women at high risk. We propose that epithelial cells isolated from breast milk of lactating women scheduled to have a breast biopsy provide a resource for assessing early changes in DNA related to increased breast cancer risk. Two hundred and fifty breast milk samples from women who are scheduled to have, or have had a breast biopsy will be collected. Of these women, we expect that roughly ten percent will be diagnosed with breast cancer. Breast milk will be collected from both the biopsied and contralateral breast of each participant. To date, 153 participants have donated breast milk, yielding 300 breast milk samples with an average size of 50 ml (±30 ml) per breast. After collection, the epithelial enriched and epithelial depleted cells from each breast were isolated. Subsequent DNA isolation of both the epithelial enriched and depleted fraction of cells yielded an average of 372 ng (range 12-5000 ng) of high quality DNA. Using the DNA extracted from epithelial enriched and depleted fractions of cells isolated from breast milk, the promoter methylation level of 9 genes known to be methylated in breast cancer will be examined using pyrosequencing technology. We will assess the promoter methylation levels of: CDH1, DAP-K, ER-α, GSTP1, HIN1, P16, RAR-ß, RASSF1A, and SFRP1 in the epithelial enriched and depleted bisulfite treated DNA of each breast milk sample. Preliminary pyrosequencing assays conducted in our lab have shown that there is a difference in RASSF1A methylation levels between the epithelial enriched DNA from the cancerous breast compared to the epithelial depleted DNA from the non-cancerous breast of the same participant. Further methylation analysis may prove that breast milk can be used as non-invasive alternative to ductal lavage or fine needle aspirate, which may yield accurate breast cancer risk assessment. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3764.

Ninety-four renal allograft recipients receiving cyclosporin A (CsA) immunosuppression for up to 4 years were examined for the presence of viral warts, keratoses, and skin cancers. They were compared with a group of 68 recipients on azathioprine who had been matched for duration of immunosuppression and other factors that might influence the occurrence of these lesions. No difference in prevalence of these tumours was found. Viral, bacterial, and fungal infections and other disorders of the skin related to immunosuppression were also noted. Apart from hypertrichosis, which occurred more frequently in the CsA group, no differences were observed. In view of the importance of duration of immunosuppression, the relative effects on the skin of the two drugs will not become apparent until CsA has been in general use for a much longer period of time. In the early stages, however, there appear to be no differences in the dermatological side-effects between CsA- and azathioprine-treated patients.

Summary— A group of 128 consecutive patients was identified on whom renal isotope studies had been performed during the first 2 months after renal transplantation and within 7 days of transplant biopsy. The prospective renogram and biopsy reports were reviewed and graded into 4 categories: severe rejection, predominant rejection, predominant acute tubular necrosis (ATN) and pure ATN. Two extreme patterns of renogram were identified: a sharp rise with a fast decline in the first min, attributed to ATN, and a slowly rising curve with no early peak occurring in severe rejection although not specific to this condition. There was a continuous intermediate spectrum. There was no inter-observer variation in gradings at the 2 ends of the spectrum. In the middle part the difference between 2 independent observers never exceeded more than 1 grade. There was good correlation between the biopsy and renogram gradings, with a discrepancy of more than 1 grade in only 5 patients; 2 of these, with severe rejection on the renogram, showed predominant ATN on biopsy, but the final clinical diagnosis was severe rejection (false positive biopsies). Two patients with biopsies showing severe rejection had a sharp initial up-slope in the renograms but a slower down-slope (over 4 min compared with 1 min in true ATN). With better definition of the criteria these renograms would not have been graded as ATN. There was 1 patient in whom no satisfactory explanation for the discrepancy was found (presumed false positive renogram). When properly defined criteria are used to interpret renograms, this simple test is at least as reliable as renal biopsy in differentiating ATN from rejection in the early post-transplant period, especially in the presence of anuria or severe oligurea.

We analyze a 140-year series of smallpox deaths in the Åland Islands, Finland. Vaccination, introduced in 1805, dramatically reduced the annual number of smallpox deaths. It also influenced the age distribution of smallpox deaths, changing smallpox from a childhood disease before 1805 to one which affected both adults and children after 1805. This appears to be due to the fact that Ålanders were usually vaccinated only once during childhood and often lost their immunity during adulthood. Spectral analysis of the prevaccination time series of smallpox deaths demonstrates a strong seven-year periodicity, reflecting the amount of time necessary to build up a cohort of nonimmune individuals. After the introduction of vaccination, the periodicity changes to eight years. The probability that a parish in Åland was affected by a smallpox epidemic is shown to be highly correlated with migration patterns and parish population sizes.

The fall in blood base occurring after the administration of secretin and the aspiration of duodenal contents was assessed in normal patients and in patients with chronic pancreatitis. In contrast to normal patients, those with chronic pancreatitis showed only a small fall or no fall in blood base level. There was no correlation between the amount of bicarbonate as determined by aspiration and that calculated from the blood base change. In 3 normal patients the magnitude of the renal compensatory mechanism in the post-secretin hour was found to be insignificant.

In 30 patients with acute myocardial infarction complicated by either advanced atrioventricular or bundle branch block, pulmonary arterial wedge pressures were studied in relation to clinical and radiographic features of left ventricular failure, changes in central venous and pulmonary arterial pressures, and arterial hypoxemia. An increase in mean pulmonary arterial wedge pressure was poorly correlated with the presence of dyspnea, audible third or fourth heart sounds, or persistent basal rales; however, it was significantly related to radiographic evidence of pulmonary venous dilatation or edema, a systolic blood pressure of less than 100 mm Hg, gallop rhythm associated with tachycardia, and heart rate in patients with sinus rhythm and normal atrioventricular conduction. Increased mean right atrial pressure indicated increased mean pulmonary arterial wedge pressure in patients with isolated anterior infarction, but in several patients with isolated inferior infarction the level of right atrial pressure was disproportionately high. The pulmonary arterial diastolic pressure closely reflected the mean pulmonary arterial wedge pressure in the absence of obstructive airways disease or pulmonary embolism. The level of the arterial oxygen tension fell as the mean pulmonary arterial wedge pressure rose, but it was considerably reduced in some patients with only moderately increased pressures. However, the latter usually had radiographic evidence of pulmonary venous dilatation or edema, suggesting that the pressure had been elevated previously to higher levels.

A patient who developed chronic renal failure secondary to the haemolytic uraemic syndrome subsequently developed life threatening microangiopathic haemolytic anaemia following renal transplantation. Transplant nephrectomy was necessary to prevent the progression ofthrombocytopenia and associated pulmonary haemorrhage.

In this randomised, double-blind, crossover trial, the efficacy in hypertension of atenolol and nifedipine as single agents or in combination was compared. 81 patients with mild to moderate essential hypertension (sitting diastolic blood pressure 100–120mm Hg, aged 20–70 years) from 6 outpatient clinics entered the study. By use of a Latin-square design, patients received, in randomised fashion, sustained release nifedipine 20mg twice daily, atenolol 50mg in the morning and then placebo in the evening, or sustained release nifedipine 20mg plus atenolol 50mg in the morning and then placebo in the evening. Each schedule was followed for 4 weeks. All treatments lowered systolic and diastolic blood pressure in the supine and standing positions compared with pretreatment values. The combination regimen significantly reduced supine and standing systolic (p < 0.01 and p < 0.001, respectively) and diastolic (p < 0.001) blood pressure compared with nifedipine alone, and it also significantly reduced supine and standing systolic (p < 0.01 and p < 0.03, respectively) and diastolic (p < 0.01) blood pressure compared with atenolol alone. Heart rate was significantly decreased by atenolol and the combination compared with nifedipine alone. 15 patients withdrew because of side effects: 9 during nifedipine treatment, 2 during atenolol treatment and 4 during combination treatment. Side effects were typical of those associated with nifedipine or atenolol. Flushes and hot sweats, which were frequent with nifedipine, were significantly less (p < 0.001) with atenolol or the combination. For selected patients who do not respond to monotherapy with nifedipine or atenolol, the combination regimen seems an appropriate and useful alternative to β-blocker/diuretic combinations, especially where ischaemic heart disease may co-exist.

Urinary concentrations of the platelet-specific protein beta-thromboglobulin (beta-TG) were measured in patients with essential hypertension. Values obtained for patients with normal renal function fell within the same population as those for a control group of normal individuals, but nevertheless 21% of these patients had urinary beta-TG values above the upper limit of the normal range. Values for patients with renal insufficiency were significantly different from the other groups, and 44% of these individuals had abnormally elevated urinary beta-TG concentrations. These elevated values may reflect an increased plasma concentration of beta-TG in patients with renal impairment or they could be an indicator of renal damage.

The indications for transplantation, complications, management and results in 127 consecutive patients who had received renal transplants between October 1960 and December 1974 are presented, In 2 cases the donor was an identical twin, in 22 a living relative other than a twin, in 2 an unrelated individual in whom nephrectomy was performed for therapeutic reasons and in 101 a cadaver. At the end of the period under review there were 37 patients alive with grafts which had functioned for 6 months or more; of these, 28 had survived for more than 2 years, 13 for more than 5 years, 6 for more than 8 years and 2 for more than 12 years. Thirty-four patients were employed or doing routine housework or receiving full-time education. One patient fathered a child 4 years after transplantation; another gave birth to a healthy infant 3 years afrer transplantation.

Comparaison de 45 malades porteurs d'un rein transplante fonctionnant bien (1 a 14 ans) et 100 malades traites par dialyse (1 a 21 ans). La maladie kystique acquise est observee dans les reins natifs de 18 des malades porteurs d'un greffon (jamais dans le greffon) et chez 63 des malades traites par dialyse. La maladie polykystique acquise progresse au cours de la dialyse et non apres transplantation

Summary When sarcoidosis of the renal parenchyma occurs it is rarely of major clinical significance. The clinical features and post-mortem findings of a case of subacute uraemic syndrome due to severe granulomatous involvement of the kidney by sarcoidosis are described. The case is unusual in that granulomatous sarcoid nephropathy resulted in death from renal failure within 4 months of onset of symptoms in the absence of clinically apparent sarcoidosis.

Urinary concentrations of the platelet-specific protein β-thromboglobulin (β-TG) were measured in patients with essential hypertension. Values obtained for patients with normal renal function fell within the same population as those for a control group of normal individuals, but nevertheless 21% of these patients had urinary β-TG values above, the upper limit of the normal range. Values for patients with renal insufficiency were significantly different from the other groups, and 44% of these individuals had abnormally elevated urinary β-TG concentrations. These elevated values may reflect an increased plasma concentration of β-TG in patients with renal impairment or they could be an indicator of renal damage.

The hemodynamic changes associated with complete heart block complicating acute myocardial infarction and the effects of artificial pacing at various rates on these measurements were studied in 13 patients. Studies were performed on the first day of complete block in all patients and were repeated on subsequent days in six. With two exceptions cardiac output was increased by pacing. In patients paced at progressively increasing ventricular rates, the rate associated with the maximal cardiac output exceeded 100/min, with one exception. However, an adequate response usually occurred between 80 and 90/min. Stroke volume fell as the rate was increased with pacing in most patients, but rose in some, suggesting improved myocardial performance. Systemic blood pressure increased, and the change paralleled cardiac output changes fairly closely as pacing rates were increased. The blood pressure did not, however, reflect reliably the absolute level of cardiac output either in heart block or after the return of sinus rhythm. Systemic vascular resistance was high in most patients with low cardiac outputs and fell with pacing at maximal output rates. There was no consistent change in mean right atrial or mean pulmonary arterial pressure with pacing during the period of study. Tension time index was low in heart block, and increased considerably with pacing, implying a corresponding increase in myocardial oxygen requirements. Pacing improved signs of depressed mental function and poor skin circulation which were invariably associated with a severe reduction in cardiac output. These signs were found to be superior to heart rate and blood pressure in clinical evaluation of the adequacy of the cardiac output in heart block and the response to pacing.

Summary Hypertension and polycythaemia rubra vera commonly occur in the same patient. This is regarded as coincidence and the hypertension does not respond to correction of polycythaemia. We report a case of renal artery stenosis occurring simultaneously with polycythaemia rubra vera and suggest that renovascular occlusive disease should be excluded in such patients with refractory hypertension.

To the Editor.— Lichen sclerosus et atrophicus (LSA) is a skin disease that begins as papules that may coalesce to form plaques and eventually become atrophic white spots. 1 Although reports of verrucous plaques have been noted, 2 this is uncommon. We report the following case to emphasize this unusual clinical presentation and to document the occurrence in a vaccination site. Report of a Case.— A 13-year-old girl was seen on May 21, 1975, for evaluation of her vaccination site. She had been vaccinated at age 2 years and revaccinated at age 6 years. The initial lesion appeared a few months following revaccination and remained unchanged until two years prior to evaluation, when it enlarged and several discrete but similar new lesions appeared. Physical Examination.— Four discrete, elevated, verrucous hyperpigmented plaques were distributed over the left deltoid area. The largest annular lesion measured 3 cm in diameter and surrounded an

Excerpt To the editor: I would like to draw attention to a possible relation between Feldman and Hershfield's paper (1) and a recent paper by Schiefer, Gee, and Ward (2). Both deal with a peculiar ...

A 49-year-old man with liver disease developed a unilateral telangiectatic lesion over the left trigeminal and left C3, C4, C5, and T1 dermatomal areas. This case fulfills the criteria for unilateral nevoid telangiectatic syndrome (UNTS), and as such represents the 11th reported case of UNTS and the first case, to our knowledge, of visceral involvement associated with a similar vascular malformation. A hemorrhagic tendency is present in this syndrome when there is gastrointestinal involvement.

To the Editor.— Penile nodulocystic lesions are rarely mentioned in standard dermatologic texts. Reports of the occurence of such lesions are also rare in the dermatologic literature. The purpose of this report is to describe a patient with a urethral diverticulum, an entity that to our knowledge has not been described previously in recent dermatologic literature. Report of a Case.— A 16-year-old uncircumcised youth came to the Dermatology Clinic for evaluation of a urethral discharge of three day's duration and a penile mass. The penile lesion was said to have been present for approximately three years. It did not change in size with micturition, although the patient did have symptoms of recurrent mild urethritis. The patient denied having had previous urologic instrumentation or neurologic dysfunction. Physical examination showed there was a yellow-green, purulent urethral discharge. A Gram stain of the discharge disclosed numerous Gramnegative intracellular diplococci compatible with Neisseria gonorrhoeae