Dorothy Jane Huang

Impact of fetal–maternal microchimerism on women's health—a review

Microchimerism is defined by the presence of circulating cells, bi-directionally transferred from one genetically distinct individual to another. It occurs either physiologically during pregnancy, or iatrogenically after blood transfusion and organ transplants. The migrated cells may persist for decades. Much controversy exists around the role of microchimeric cells in the pathogenesis of various diseases and around their role in tissue repair. Microchimerism has been investigated in different autoimmune disorders, such as systemic sclerosis, systemic lupus erythematosus, autoimmune thyroid diseases, primary biliary cirrhosis and juvenile inflammatory myopathies. Recent data have demonstrated the promising role of microchimeric cells in the maternal response to tissue injuries by differentiating into many lineages. Therefore, further understanding of fetal–maternal microchimerism may help in anticipating its implications in disease as well as in more general womens health issues.

Metastatic patterns at autopsy in patients with ovarian carcinoma.

Previously published studies concerning autopsy findings in ovarian cancer failed to consider the broad differences in factors that influence the course of disease. Furthermore, those studies were conducted when the currently accepted standards in diagnostics and therapy had not been fully established. The objective of the current study was to determine the frequency and sites of metastases in patients with ovarian cancer with particular attention to the clinical course and therapy.

Isolation of Cell-Free DNA from Maternal Plasma Using Manual and Automated Systems

Cell-free fetal DNA present in the maternal circulation holds great potential for noninvasive prenatal diagnosis and analysis of fetal genetic traits. However, only approximately 3-6% of total DNA in the maternal plasma is of fetal origin. Because of its scarcity in the maternal circulation, various methods have been developed and tested to optimize the extraction of this rare material from plasma. Here, we first describe the commonly used protocol for separating plasma from whole blood samples. We also describe two commercially available methods for the extraction of cell-free DNA from maternal plasma, which we have found particularly straightforward and easy to use: a manual method using the High Pure PCR Template Preparation kit (Roche Diagnostics) and an automated system using the MagNA Pure LC instrument (Roche Diagnostics). Use of the methods described here will help to ensure maximum yield and purity of cell-free fetal DNA extracted from maternal plasma samples for downstream analyses.

Breast cancer in women 80 years of age and older: A comprehensive analysis of an underreported entity

Abstract Background. Approximately 10% of breast cancer (BC) patients are over the age of 80. We present the first comprehensive review on this particular group of patients. Patients and methods. The treatments and disease courses of an unselected cohort of patients, whose age at first diagnosis was ≥ 80 years (n = 151), were compared to those of a group of women, who were aged 56–66 years (n = 372). Results. The group of elderly patients had larger tumors at first diagnosis (25 mm vs. 18 mm, p < 0.001) and higher disease stages (I: 31.1% vs. 44.1%, IV: 11.9% vs. 5.4%; each p < 0.001). There were no significant differences between both groups in terms of histologic subtype, grading, hormonal receptor status and HER2 status. The tumors of older patients were more often detected by clinical examination (38.9% vs. 17.0%, p < 0.001) and less often by mammography/sonography (10.4% vs. 29.9%, p < 0.001). The rate of patients who died of BC were similar in both groups (21.2% vs. 21.5%, p = 1.00). In the patients who had no evidence of metastases and who opted for primary non-surgical management (n = 21), the tumor could be stabilized without considerable morbidity in only 42.9%. Persistence to adjuvant endocrine therapy was comparable (83.0% vs. 88.3%, p = 0.357). In the adjuvant as well as in the palliative settings, elderly patients received less chemotherapy than younger ones (adjuvant: 1.6% vs. 23.3%; palliative: 32.3% vs. 68.4%; each p < 0.001). For palliative treatments only, elderly patients received fewer treatment regimens (≥ 3 therapy lines: 16.0% vs. 54.9%, p < 0.001). In those patients who died of BC, elderly women had inferior overall (25 vs. 54.5 months, p < 0.001) as well as metastatic-disease survival (11.5 vs. 19 months, p = 0.062). Conclusion. It must be ensured that appropriate standard therapies should not be routinely withheld in older patients based on erroneous perceptions regarding the biological nature of BC in the elderly and lack of knowledge about available therapy regimens. Physicians should consider that preservation of current life circumstances and maintenance of quality of life are frequently more important than “classical” hard medical facts such as survival times.

Isolation of cell-free RNA from maternal plasma

The discovery of cell-free RNA in plasma and serum samples provides possibilities for noninvasive prenatal diagnosis. Quantitative alterations of cell-free placental-derived mRNA in the maternal circulation are associated with many pregnancy-related disorders, such as preeclampsia and preterm labor. Obtaining circulating cell-free RNA is the first and often the most critical step for analyzing the placental-derived mRNA in maternal blood. We have compared different protocols for the extraction of cell-free RNA from plasma samples, and we have found the protocol using TRIzol LS reagent (Invitrogen) as lysis buffer combined with RNeasy Mini kit (QIAGEN) to be the optimal method for extracting high-quality cell-free RNA in the highest quantities possible. This method is also amenable to the simultaneous extraction of cell-free RNA from many different samples, by use of the QIAGEN Vacuum Manifold QIAvac 24 Plus.

Reliable detection of trisomy 21 using MALDI-TOF mass spectrometry

Purpose: Current diagnostic methods for chromosomal abnormalities rely mainly on karyotyping and occasionally fluorescent in situ hybridization or quantitative polymerase chain reaction. We describe an alternative molecular method for the detection of trisomy 21 involving mass spectrometric analysis of single nucleotide polymorphisms.Methods: In collaboration with Sequenom, Inc., 350 blinded amniotic fluid, amniocyte culture, chorionic villus, or amniotic fluid supernatant samples were analyzed for trisomy 21 using SNP analysis and matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. Peak ratios were calculated for heterozygous genotypes and compared to control values generated from known euploid samples. An analytical algorithm using standard deviations from control values was used to determine the probability of a sample being affected or unaffected.Results: Seventy-three trisomy 21 samples from among the 350 blinded samples were correctly identified. There were no false-positive or false-negative results among the complete trisomy 21 samples. One sample exhibiting mosaicism for trisomy 21 was identified as being unaffected.Conclusions: MALDI-TOF mass spectrometry is a robust and reproducible method for the detection of trisomy 21. Its amenability to high-throughput analysis and high degree of multiplexing make it a potential future diagnostic tool for the detection of other aneuploidies as well.

Lifestyle and health-related predictors of cervical cancer screening attendance in a Swiss population-based study

BACKGROUND Since the implementation of cervical cancer (CC) screening, incidence and mortality rates have decreased worldwide. Little is known about lifestyle and health-related predictors of cervical cancer screening attendance in Switzerland. Our aim was to examine the relationship between lifestyle and health-related factors and the attendance to CC screening in Switzerland. METHODS We analyzed data of 20-69 years old women (n=7319) of the Swiss Health Survey (SHS) 2012. Lifestyle factors included body mass index, smoking status, alcohol consumption, physical activity and attention to diet. Health-related factors of interest were diabetes, hypertension, high cholesterol levels, chronic diseases, self-perceived health, and psychological distress. We performed multivariable logistic regression analyses with the dichotomized CC screening status as outcome measure and adjusted for demographic factors. RESULTS Obesity, low physical activity, and not paying attention to diet were statistically significantly associated with lower CC screening participation. High cholesterol levels and history of chronic diseases were statistically significantly positively associated with screening participation. CONCLUSION Being obese, physically inactive and non-attention to diet are risk factors for CC screening attendance. These findings are of importance for improving the CC screening practices of low-user groups.

Cannabinoid hyperemesis syndrome: an underreported entity causing nausea and vomiting of pregnancy

IntroductionIn the western world, cannabis is the most widely used drug of abuse. Cannabinoid hyperemesis syndrome, which seems to be a rare paradoxical reaction in individuals with a particular predisposition, is characterized by cyclic severe nausea and vomiting in long-term cannabis users. While the symptoms are unresponsive to antiemetic drugs, compulsive hot baths result in a considerable symptom relief.MethodsWe report the first case of cannabinoid hyperemesis syndrome in pregnancy. A 26-year-old patient was admitted to our clinic in the 10th week of gestation.ConclusionBefore undertaking time-consuming and expensive medical examinations to rule out other medical reasons for therapy-resistant hyperemesis in pregnancy, obstetricians should determine whether compulsive bathing or showering provides symptomatic relief and ask specific questions regarding possible/suspected cannabis consumption.

Distant Metastatic Breast Cancer as an Incurable Disease: A Tenet With a Need for Revision

Purpose:Published reports provide level-III evidence in support of the hypothesis that distant metastatic breast cancer (MBC) might be curable in up to 3% of cases through a multidisciplinary approach including combination chemotherapy regimens in selected patients, usually young, and with limited metastases. Our study evaluates the rate and characteristics of long-term survivors based on a nonselective study cohort. Patients and Methods:We analyzed the data from 149 patients in whom distant MBC was diagnosed from 1990 to 1999. Results:Five patients (3.4%) were long-term survivors (9-14 years after initial diagnosis of MBC) without any clinical evidence of disease. They had a 2-peaked distribution of age: 3 were 41-57 years old at the diagnosis of MBC and 2 were much older (76, 79 years). Median survival time after diagnosis of MBC was 152 (range, 109-172) months. Three patients had isolated metastatic lesions, although 1 patient had multiple organ metastases and another extensive bone metastases. In 4 of 5 cases, long-term survival was achieved without the administration of chemotherapy. Discussion:Long-term survivors in MBC comprise a relatively heterogeneous group, and the factors which lead to the quite rare situation of long-term survival can hardly be evaluated systematically. Aggressive chemotherapy regimens appear not to be a key factor for survival. Furthermore, in a nonselective study cohort, some patients clearly are not only alive but also disease-free more than 12 years after initial relapse. This fraction may be small, but the chance for survival, and even for cure, truly exists.

Maternal smoking: effect on circulating cell-free fetal and total DNA levels in maternal plasma from the second trimester.

OBJECTIVE: To estimate whether potential clinical applications of cell-free fetal and total DNA in the field of noninvasive prenatal diagnosis need to be adjusted for maternal smoking status. METHODS: In this study, using 344 maternal blood samples from the second trimester of pregnancy, circulating cell-free DNA in maternal plasma samples, specific for the SRY and DYS14 loci (representing fetal DNA) and GAPDH sequence (representing total genomic DNA) were quantified by real-time polymerase chain reaction. RESULTS: Fetal sex determination was 100% accurate using a combination of probes for SRY and DYS14. The levels of DYS14 and SRY detected were significantly correlated (r=0.884, P<.001). No significant difference was seen between the quantitative levels of cell-free male fetal DNA between the smoking groups and control group. Similarly, no significant difference was seen in the amount of total cell-free DNA in the study population. CONCLUSION: In contrast to first- and second-trimester screening assays for Down syndrome, where smoking status significantly affect levels of maternal serum analytes, smoking status does not affect quantitative levels of cell-free fetal DNA or total cell-free DNA in maternal plasma. LEVEL OF EVIDENCE: II