Novel Strategies in Cancer Prevention and Fertility Preservation with Tamoxifen

Women at high risk for breast cancer are often also at high risk for ovarian cancer, reflecting similar risk factors and suggesting intertwined disease pathways and common prevention targets. A novel strategy to overcome obstacles in preventing ovarian neoplasia (low incidence, lack of specific disease markers, and difficulties in tissue sampling), the deadliest gynecologic cancer, may be to develop a prevention strategy that targets breast and ovarian cancer simultaneously. Tamoxifen, a selective estrogen receptor modulator, reduces hormone responsive breast cancer risk by 50% but its effects on risk of ovarian cancer, also hormonal responsive, are unclear. The goals of this work were to 1) develop and characterize a preclinical model of concurrent breast and ovarian cancer and 2) use this dual cancer model to examine the efficacy of tamoxifen to prevent both breast and ovarian cancer. Mammary carcinogens [7,12-dimethylbenz[α]anthracene (DMBA), N-methyl-N-nitrosourea and estradiol (Ey2)] were tested separately in combination with local ovarian DMBA administration to determine the best combined treatment to induce mammary and ovarian cancer concurrently and effectively in the rat. Results showed that systemic Ey2 and ovarian DMBA promoted the highest incidence of dysplasia in the mammary gland and ovary and elevated levels of mammary Ki-67 and cyclooxygenase 2 (COX-2) mimicking the human disease. Next, the ability of tamoxifen to prevent mammary and ovarian cancer simultaneously was evaluated. Tamoxifen which inhibited mammary carcinogenesis and normalized levels of Ki-67 and COX-2, had no effect on (neither accelerated nor inhibited) ovarian cancer progression. In addition, carcinogen treatment increased levels of stem cell markers, Oct-4 and aldehyde dehydrogenase-1, in the mammary gland; interestingly, this expansion was not reversed by tamoxifen. Intriguingly, while examining ovaries from this study, we serendipitously discovered an apparent protective effect of tamoxifen against DMBA-induced follicular destruction and this effect was further investigated. Chemotherapy and environmental toxicants (e.g. DMBA) deplete ovarian follicles and often lead to accelerated ovarian aging and premature ovarian failure; however, there is no established treatment that can protect the ovary from these toxic insults. In vivo, rats were treated with tamoxifen and DMBA or cyclophosphamide (the most ovotoxic chemotherapy) and total numbers of follicles in the ovary were determined. In vitro, ovarian organ culture and oocyte culture were carried out to examine local effects of tamoxifen on DMBA-induced follicle loss and doxorubicin-induced oocyte fragmentation, respectively. We demonstrated for the first time that tamoxifen protects ovarian follicles against not only DMBA- but also chemotherapy (cyclophosphamide and doxorubicin)-induced ovarian damage. Clinically, tamoxifen has already been tested for safe use as an adjuvant therapy for several cancers; therefore, if translated into clinical use, these results may have immediate impact on options for fertility preservation and quality of life in young female cancer patients undergoing chemotherapy. The long term goals of this work are to 1) use the dual cancer model to screen for promising agents that decrease risks for both breast and ovarian cancer and 2) examine the mechanism by which tamoxifen inhibits toxicant-induced ovarian follicle loss

Healthcare use by children and young adults with cerebral palsy

Aim To link routinely collected health data to a cerebral palsy (CP) register in order to enable analysis of healthcare use by severity of CP. Method The Northern Ireland Cerebral Palsy Register was linked to hospital data. Data for those on the CP register born between 1st January 1981 and 31st December 2009 and alive in 2004 were extracted, forming a CP cohort (n=1684; 57% males, 43% females; aged 0–24y). Frequencies of healthcare events, and the reasons for them, were reported according to CP severity and compared with those without CP who had had at least one hospital attendance in Northern Ireland within the study period. Results Cases of CP represented 0.3% of the Northern Ireland population aged 0 to 24 years but accounted for 1.6% of hospital admissions and 1.6% of outpatient appointments. They had higher rates of elective admissions and multi‐day hospital stays than the general population. Respiratory conditions were the most common reason for emergency admissions. Those with most severe CP were 10 times more likely to be admitted, and four times more likely to attend outpatients, than those with mild CP. Interpretation Linkage between a register and routinely collected healthcare data provided a confirmed cohort of cases of CP that was sufficiently detailed to analyse healthcare use by disease severity

Validating coding for a chronic condition (Cerebral Palsy) in routinely collected hospital admissions data

Introduction Data-linkage provides an opportunity to evaluate healthcare utilisation of patients with chronic health conditions. However the consistency and accuracy of recording chronic conditions in routinely collected healthcare data is unclear and we aim to explore this in children and young people (CYP) with a cerebral palsy (CP). Objectives and Approach The study aims to validate CP case identification in routinely collected data by data-linking cases from The Northern Ireland Cerebral Palsy Register (NICPR) born 1981-2011 with routinely collected hospital admission data and cross-referencing cases between the two datasets. CP cases were identified in the administrative data using ICD-10 codes G80 (cerebral palsy), G81 (hemiplegia), G82 (paraplegia and tetraplegia) and G83.0-83.3 (diplegia and monoplegias). Results There were 1,693 cases in the NICPR cohort and 1,733 cases of CP identified in the hospital admissions data, 915 of which were on the NICPR giving a positive predictive value for identifying a CP case within routinely collected data of 52.8%. Specificity was high (99.8%) however sensitivity was low (54.1%). 1,157 (68.3%) patients on the NICPR had at least one hospital admission. 914 (79.0 %) cases were coded at some time point with G80-83.3 (829 (71.7%) G80, 328 (28.3%)  G81-83.3). NICPR cases had a total 11,844 hospital admissions and a CP code was recorded during 6,397 (54.0%). The type of CP was recorded in NICPR for 1,673 (98.8%) cases whereas in the hospital admissions, data the majority (70.6%) were coded as other/unspecified CP (G80.8-9). Conclusion/Implications This study adds to understanding of CP coding practices within routinely collected hospital data, further supporting claims that coding of CP within such datasets is poor. Coding must be accurate and consistent if data are to be meaningful, comparable and useful to inform health outcome reviews and patient care

Polarized Secretion of Interleukin (IL)-6 and IL-8 by Human Airway Epithelia 16HBE14o- Cells in Response to Cationic Polypeptide Challenge

BACKGROUND: The airway epithelium participates in asthmatic inflammation in many ways. Target cells of the epithelium can respond to a variety of inflammatory mediators and cytokines. Damage to the surface epithelium occurs following the secretion of eosinophil-derived, highly toxic cationic proteins. Moreover, the surface epithelium itself is responsible for the synthesis and release of cytokines that cause the selective recruitment, retention, and accumulation of various inflammatory cells. To mimic the damage seen during asthmatic inflammation, the bronchial epithelium can be challenged with highly charged cationic polypeptides such as poly-L-arginine. METHODOLOGY/PRINCIPAL FINDINGS: In this study, human bronchial epithelial cells, 16HBE14o- cells, were "chemically injured" by exposing them to poly-l-arginine as a surrogate of the eosinophil cationic protein. Cytokine antibody array data showed that seven inflammatory mediators were elevated out of the 40 tested, including marked elevation in interleukin (IL)-6 and IL-8 secretion. IL-6 and IL-8 mRNA expression levels were elevated as measured with real-time PCR. Cell culture supernatants from apical and basolateral compartments were collected, and the IL-6 and IL-8 production was quantified with ELISA. IL-6 and IL-8 secretion by 16HBE14o- epithelia into the apical compartment was significantly higher than that from the basolateral compartment. Using specific inhibitors, the production of IL-6 and IL-8 was found to be dependent on p38 MAPK, ERK1/2 MAPK, and NF-kappaB pathways. CONCLUSIONS/SIGNIFICANCE: The results clearly demonstrate that damage to the bronchial epithelia by poly-L-arginine stimulates polarized IL-6 and IL-8 secretion. This apically directed secretion of cytokines may play an important role in orchestrating epithelial cell responses to inflammation

The Assembly History of Field Spheroidals: Evolution of Mass-to-light Ratios and Signatures of Recent Star Formation

We present a comprehensive catalog of high signal-to-noise spectra obtained with the DEIMOS spectrograph on the Keck II telescope for a sample of F850LP<22.43 (AB) field spheroidal (E+S0s; 163) and bulge dominated disk (61) galaxies in the redshift range 0.2<z<1.2. We examine the zero point, tilt and scatter of the Fundamental Plane (FP) as a function of redshift and morphological properties, carefully accounting for luminosity-dependent biases via Montecarlo simulations. The evolution of the overall FP can be represented by a mean change in effective mass-to-light ratio given by <d \log (M/L_{\rm B})/dz>=-0.72^{+0.07}_{-0.05}\pm0.04. However, this evolution depends significantly on the dynamical mass, being slower for larger masses as reported in a previous letter. In addition, we separately show the intrinsic scatter of the FP increases with redshift as d(rms(M/L_{\rm B}))/dz=0.040\pm0.015. Although these trends are consistent with single burst populations which formed at $z_f>2$ for high mass spheroidals and z_{f}~1.2 for lower mass systems, a more realistic picture is that most of the stellar mass formed in all systems at z>2 with subsequent activity continuing to lower redshifts (z<1.2). The fraction of stellar mass formed at recent times depend strongly on galactic mass, ranging from <1% for masses above 10^{11.5} M_{\odot} to 20-40% below 10^{11} M_{\odot}. Independent support for recent activity is provided by spectroscopic ([\ion{O}{2}] emission, H\delta) and photometric (blue cores and broad-band colors) diagnostics. Via the analysis of a large sample with many independent diagnostics, we are able to reconcile previously disparate interpretations of the assembly history of field spheroidals. [Abridged]Comment: 26 pages including 24 figures, submitted to ApJ. Complete and compact version with full resolution images available at http://www.astro.ucla.edu/~ttreu/ms.pd

Functional Specialization of Cellulose Synthase Isoforms in a Moss Shows Parallels with Seed Plants

The secondary cell walls of tracheary elements and fibers are rich in cellulose microfibrils that are helically oriented and laterally aggregated. Support cells within the leaf midribs of mosses deposit cellulose-rich secondary cell walls, but their biosynthesis and microfibril organization have not been examined. Although the Cellulose Synthase (CESA) gene families of mosses and seed plants diversified independently, CESA knockout analysis in the moss Physcomitrella patens revealed parallels with Arabidopsis (Arabidopsis thaliana) in CESA functional specialization, with roles for both subfunctionalization and neofunctionalization. The similarities include regulatory uncoupling of the CESAs that synthesize primary and secondary cell walls, a requirement for two or more functionally distinct CESA isoforms for secondary cell wall synthesis, interchangeability of some primary and secondary CESAs, and some CESA redundancy. The cellulose-deficient midribs of ppcesa3/8 knockouts provided negative controls for the structural characterization of stereid secondary cell walls in wild type P. patens. Sum frequency generation spectra collected from midribs were consistent with cellulose microfibril aggregation, and polarization microscopy revealed helical microfibril orientation only in wild type leaves. Thus, stereid secondary walls are structurally distinct from primary cell walls, and they share structural characteristics with the secondary walls of tracheary elements and fibers. We propose a mechanism for the convergent evolution of secondary walls in which the deposition of aggregated and helically oriented microfibrils is coupled to rapid and highly localized cellulose synthesis enabled by regulatory uncoupling from primary wall synthesis

Identifying cerebral palsy from routinely-collected data in England and Wales

Purpose: An observational study using routinely-collected health care data to describe the extent to which children and young people (CYP) with cerebral palsy (CP) can be identified and the prevalence of CP can be estimated. Patients and methods: Routinely-collected anonymized data, for CYP (aged 0–25 years old between 1 January 2004 and 31 December 2014) were analyzed in two linked datasets, from England and Wales respectively. Datasets included National Health Service; General Practitioner (GP), inpatients, outpatients, and national mortality records. CP was identified using ICD-10 codes G80.0–G83.3 and equivalent Read v2 codes. Ascertainment rates of CP were identified for each data source and compared between countries. Frequency and consistency of coding were investigated, and prevalence of CP estimated. Results: A total of 7,113 and 5,218 CYP with CP were identified in the English and Welsh datasets respectively. Whilst the majority of CYP with CP would be expected to attend their GP, 65.3% (4,646/7,113) of English and 65.1% (3,396/5,218) of Welsh cases were ascertained from GP datasets. Further cases were identified solely in inpatient datasets (2,410 in England, 1,813 in Wales). Few cases were coded for CP within outpatient datasets. Four character codes that specified CP type were rarely used; one in five health care records were coded both with G80 codes (explicitly CP) and with G81–83 codes (other paralytic syndromes) or equivalent Read codes. Estimated period prevalence of CYP with CP was 2.5–3.4 per 1,000 in England and 2.4–3.2 per 1,000 in Wales. Conclusion: In England and Wales, coding of CP in routine data is infrequent, inconsistent, non-specific, and difficult to isolate from conditions with similar physical signs. Yet the prevalence estimates of CP were similar to those reported elsewhere. To optimize case recognition we recommend improved coding quality and the use of both primary and secondary care datasets as a minimum

Developmental conversion of thymocyte-attracting cells into self-antigen-displaying cells in embryonic thymus medulla epithelium

Thymus medulla epithelium establishes immune self-tolerance and comprises diverse cellular subsets. Functionally relevant medullary thymic epithelial cells (mTECs) include a self-antigen-displaying subset that exhibits genome-wide promiscuous gene expression promoted by the nuclear protein Aire and that resembles a mosaic of extrathymic cells including mucosal tuft cells. An additional mTEC subset produces the chemokine CCL21, thereby attracting positively selected thymocytes from the cortex to the medulla. Both self-antigen-displaying and thymocyte-attracting mTEC subsets are essential for self-tolerance. Here, we identify a developmental pathway by which mTECs gain their diversity in functionally distinct subsets. We show that CCL21-expressing mTECs arise early during thymus ontogeny in mice. Fate-mapping analysis reveals that self-antigen-displaying mTECs, including Aire-expressing mTECs and thymic tuft cells, are derived from CCL21-expressing cells. The differentiation capability of CCL21-expressing embryonic mTECs is verified in reaggregate thymus experiments. These results indicate that CCL21-expressing embryonic mTECs carry a developmental potential to give rise to self-antigen-displaying mTECs, revealing that the sequential conversion of thymocyte-attracting subset into self-antigen-displaying subset serves to assemble functional diversity in the thymus medulla epithelium

Primary, secondary care and mortality data-linked to identify the prevalence of cerebral palsy in children and young people

Introduction Evaluations of healthcare utilisation for children and young people (CYP) with chronic conditions, are increasingly relying upon routinely collected healthcare data to estimate healthcare burden and inform national policy and practice. However, chronic conditions are not consistently or accurately recorded making it difficult to conduct valid epidemiological analyses. Objectives and Approach We explored routinely collected healthcare datasets of 2,122,914 CYP in the English Clinical Practice Research Dataset (CPRD) and 1,636,252 CYP in the Welsh Secure Anonymous Information Linkage (SAIL) databank to identify patients with CP (an exemplar of a chronic neurodisability) from diagnosis coding (G80-83.3). Linked primary care, hospital admission outpatient and mortality data were searched from birth and populations of CYP aged 0-25 years between 2004 and 2014 with and without CP were identified. We detected the ascertainment sources and compared the results from CPRD and SAIL. In a sample of cases G80-83 codes were validated against clinical records. Results England: Some 7,500 cases of CP were identified (period prevalence: 3.5 per 1000 CYP). Of those, 36.6% were identified from hospital admissions; 20.6% from GP data, 42.0% were in both datasets with 0.8% from outpatient/ONS mortality data. Wales: Some 5,400 cases of CP were identified (period prevalence: 3.3 per 1000 CYP). Of those, 38.6% were identified from hospital admissions, 25.3% from GP data, 36.0% were in both datasets, and 0.1% from mortality/outpatient datasets. 729/877(83.1%) cases coded as G80-83 in secondary care case notes were validated cases of CP leaving 16.9% that were incorrectly coded. Approximately 70% of G80 cases were recorded as G80.8-9 (CP other/unspecified). Roughly 30% of cases were only coded as CP on one occasion within the primary and secondary care datasets. Conclusion/Implications Similar proportions of CP cases were identified in the two datasets giving similar period prevalences. Inconsistent and incorrect coding will affect the accuracy of these figures and precludes any analysis by disease type/severity. Improved coding of chronic conditions is needed before accurate healthcare analysis of routine data can be undertaken

Dual- Isolation and Profiling of Circulating Tumor Cells and Cancer Exosomes from Blood Samples with Melanoma Using Immunoaffinity- Based Microfluidic Interfaces

Melanoma is among the most aggressive cancers, and its rate of incidence continues to grow. Early detection of melanoma has been hampered due to the lack of promising markers for testing. Recent advances in liquid biopsy have proposed noninvasive alternatives for cancer diagnosis and monitoring. Circulating tumor cells (CTCs) and cancer- exosomes are gaining influence as promising biomarkers because of their cancer- associated molecular markers and signatures. However, technologies that offer the dual- isolation of CTCs and exosomes using a single sample have not been thoroughly developed. The dual- utilization OncoBean (DUO) device is conjugated with melanoma specific antibodies, MCAM and MCSP, enabling simultaneous CTC and exosome isolations. Using blood samples from patients, CTCs and exosomes are specifically isolated from a single sample and then undergo molecular profiling for comprehensive study. Melanoma patients have 0- 17CTCs mL- 1 and 299 µg exosomal protein mL- 1 while healthy donors display fewer than 2CTCs and 75.6 µg of exosomes mL- 1, respectively. It is also demonstrated that both markers express melanoma- associated genes using multiplex qRT- PCR to test for expression pattern of a 96 gene panel. The dual isolation and molecular characterization will allow for further research into melanoma to identify viable markers for disease progression and treatment efficacy.Circulating tumor cells (CTCs) and cancer exosomes are gaining influence as promising biomarkers in liquid biopsy, however, technologies that offer the dual- isolation of CTCs and exosomes have not been developed. Here, a microfluidic device is devised conjugated with melanoma cell adhesion molecule (MCAM) and melanoma- associated chondroitin sulfate proteoglycan (MCSP) for isolation and molecular profiling of both melanoma CTCs and exosomes.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/163448/3/advs1971.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/163448/2/advs1971_am.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/163448/1/advs1971-sup-0001-SuppMat.pd