22 research outputs found
Transgénicos, mirada desde la ética
El gusano de la curiosidad, ese impulso que se siente en el estómago como hambre de saber, el mismo que tienen las y los niños, el mismo que ha tenido el ser humano desde su aparición en la Tierra, es padre y madre de la ciencia. Va con el hombre y con la mujer la búsqueda de la verdad, de la realidad del mundo que nos rodea y de nuestra propia realidad. Es ése el objetivo más elevado de la ciencia y el acicate que nos hace crecer y desarrollarnos como individuos y como especie. ¿Son esos los objetivos de la biotecnología aplicada en los cultivos transgénicos
Comparison of methods for the isolation of human breast epithelial and myoepithelial cells.
Two lineages, epithelial, and myoepithelial cells are the main cell populations in the normal mammary gland and in breast cancer. Traditionally, cancer research has been performed using commercial cell lines, but primary cell cultures obtained from fresh breast tissue are a powerful tool to study more reliably new aspects of mammary gland biology, including normal and pathological conditions. Nevertheless, the methods described to date have some technical problems in terms of cell viability and yield, which hamper work with primary mammary cells. Therefore, there is a need to optimize technology for the proper isolation of epithelial and myoepithelial cells. For this reason, we compared four methods in an effort to improve the isolation and primary cell culture of different cell populations of human mammary epithelium. The samples were obtained from healthy tissue of patients who had undergone mammoplasty or mastectomy surgery. We based our approaches on previously described methods, and incorporated additional steps to ameliorate technical efficiency and increase cell survival. We determined cell growth and viability by phase-contrast images, growth curve analysis and cell yield, and identified cell-lineage specific markers by flow cytometry and immunofluorescence in 3D cell cultures. These techniques allowed us to better evaluate the functional capabilities of these two main mammary lineages, using CD227/K19 (epithelial cells) and CD10/K14 (myoepithelial cells) antigens. Our results show that slow digestion at low enzymatic concentration combined with the differential centrifugation technique is the method that best fits the main goal of the present study: protocol efficiency and cell survival yield. In summary, we propose some guidelines to establish primary mammary epithelial cell lines more efficiently and to provide us with a strong research instrument to better understand the role of different epithelial cell types in the origin of breast cancer
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Comparison of methods for the isolation of human breast epithelial and myoepithelial cells
Two lineages, epithelial, and myoepithelial cells are the main cell populations in the normal mammary gland and in breast cancer. Traditionally, cancer research has been performed using commercial cell lines, but primary cell cultures obtained from fresh breast tissue are a powerful tool to study more reliably new aspects of mammary gland biology, including normal and pathological conditions. Nevertheless, the methods described to date have some technical problems in terms of cell viability and yield, which hamper work with primary mammary cells. Therefore, there is a need to optimize technology for the proper isolation of epithelial and myoepithelial cells. For this reason, we compared four methods in an effort to improve the isolation and primary cell culture of different cell populations of human mammary epithelium. The samples were obtained from healthy tissue of patients who had undergone mammoplasty or mastectomy surgery. We based our approaches on previously described methods, and incorporated additional steps to ameliorate technical efficiency and increase cell survival. We determined cell growth and viability by phase-contrast images, growth curve analysis and cell yield, and identified cell-lineage specific markers by flow cytometry and immunofluorescence in 3D cell cultures. These techniques allowed us to better evaluate the functional capabilities of these two main mammary lineages, using CD227/K19 (epithelial cells) and CD10/K14 (myoepithelial cells) antigens. Our results show that slow digestion at low enzymatic concentration combined with the differential centrifugation technique is the method that best fits the main goal of the present study: protocol efficiency and cell survival yield. In summary, we propose some guidelines to establish primary mammary epithelial cell lines more efficiently and to provide us with a strong research instrument to better understand the role of different epithelial cell types in the origin of breast cancer
Population-based multicase-control study in common tumors in Spain (MCC-Spain): rationale and study design
Introduction: We present the protocol of a large population-based case-control study of 5 common tumors
in Spain (MCC-Spain) that evaluates environmental exposures and genetic factors.
Methods: Between 2008-2013, 10,183 persons aged 20-85 years were enrolled in 23 hospitals and primary
care centres in 12 Spanish provinces including 1,115 cases of a new diagnosis of prostate cancer,
1,750 of breast cancer, 2,171 of colorectal cancer, 492 of gastro-oesophageal cancer, 554 cases of chronic
lymphocytic leukaemia (CLL) and 4,101 population-based controls matched by frequency to cases by age,
sex and region of residence. Participation rates ranged from 57% (stomach cancer) to 87% (CLL cases) and
from 30% to 77% in controls. Participants completed a face-to-face computerized interview on sociodemographic
factors, environmental exposures, occupation, medication, lifestyle, and personal and family
medical history. In addition, participants completed a self-administered food-frequency questionnaire
and telephone interviews. Blood samples were collected from 76% of participants while saliva samples
were collected in CLL cases and participants refusing blood extractions. Clinical information was recorded
for cases and paraffin blocks and/or fresh tumor samples are available in most collaborating hospitals.
Genotyping was done through an exome array enriched with genetic markers in specific pathways. Multiple
analyses are planned to assess the association of environmental, personal and genetic risk factors
for each tumor and to identify pleiotropic effects.
Discussion: This study, conducted within the Spanish Consortium for Biomedical Research in Epidemiology
& Public Health (CIBERESP), is a unique initiative to evaluate etiological factors for common cancers
and will promote cancer research and prevention in Spain.The study was partially funded by the “Accion Transversal
del Cancer”, approved on the Spanish Ministry Council on the
11th October 2007, by the Instituto de Salud Carlos III-FEDER
(PI08/1770, PI08/0533, PI08/1359, PS09/00773, PS09/01286,
PS09/01903, PS09/02078, PS09/01662, PI11/01403, PI11/01889,
PI11/00226, PI11/01810, PI11/02213, PI12/00488, PI12/00265,
PI12/01270, PI12/00715, PI12/00150), by the Fundación Marqués
de Valdecilla (API 10/09), by the ICGC International Cancer Genome
Consortium CLL, by the Junta de Castilla y León (LE22A10-2), by
the Consejería de Salud of the Junta de Andalucía (PI-0571), by the
Conselleria de Sanitat of the Generalitat Valenciana (AP 061/10),
by the Recercaixa (2010ACUP 00310), by the Regional Government
of the Basque Country by European Commission grants FOOD-CT-
2006-036224-HIWATE, by the Spanish Association Against Cancer
(AECC) Scientific Foundation, by the The Catalan Government
DURSI grant 2009SGR1489
Primary and Secondary Immunodeficiency Diseases in Oncohaematology: Warning Signs, Diagnosis, and Management
Background: Immunodeficiencies (ID), in particular primary immunodeficiencies (PID), are often associated with haematological manifestations, such as peripheral cytopenias or lymphoproliferative syndromes. Early diagnosis and management have significant prognostic implications. Secondary immunodeficiencies (SID) may also be induced by oncohaematological diseases and their treatments. Haematologists and oncologists must therefore be aware of the association between blood disorders and cancer and ID, and be prepared to offer their patients appropriate treatment without delay. Our aim was to define the warning signs of primary and secondary IDs in paediatric and adult patients with oncohaematological manifestations.Methods: A multidisciplinary group of six experts (2 haematologists, 2 immunologists, and 2 paediatricians specializing in ID) conducted a literature review and prepared a document based on agreements reached an in-person meeting. An external group of 44 IDs specialists from all over Spain assessed the document and were consulted regarding their level of agreement.Results: This document identifies the haematological and extra-haematological diseases that should prompt a suspicion of PIDs in adults and children, in both primary care and haematology and oncology departments. Cytopenia and certain lymphoproliferative disorders are key diagnostic pointers. The diagnosis must be based on a detailed clinical history, physical exploration, complete blood count and standard laboratory tests. The immunological and haematological tests included in the diagnostic process will depend on the care level. Patients who are candidates for immunoglobulin replacement therapy must be carefully selected, and treatment should be offered as soon as possible to avoid the development of complications. Finally, this document recommends procedures for monitoring these patients.Conclusions: This document combines scientific evidence with the opinion of a broad panel of experts, and emphasizes the importance of an early diagnosis and treatment to avoid complications. The resulting document is a useful tool for primary care physicians and specialists who see both adult and paediatric patients with oncohaematological diseases
SARS-CoV-2 susceptibility and COVID-19 disease severity are associated with genetic variants affecting gene expression in a variety of tissues
Variability in SARS-CoV-2 susceptibility and COVID-19 disease severity between individuals is partly due to
genetic factors. Here, we identify 4 genomic loci with suggestive associations for SARS-CoV-2 susceptibility
and 19 for COVID-19 disease severity. Four of these 23 loci likely have an ethnicity-specific component.
Genome-wide association study (GWAS) signals in 11 loci colocalize with expression quantitative trait loci
(eQTLs) associated with the expression of 20 genes in 62 tissues/cell types (range: 1:43 tissues/gene),
including lung, brain, heart, muscle, and skin as well as the digestive system and immune system. We perform
genetic fine mapping to compute 99% credible SNP sets, which identify 10 GWAS loci that have eight or fewer
SNPs in the credible set, including three loci with one single likely causal SNP. Our study suggests that the
diverse symptoms and disease severity of COVID-19 observed between individuals is associated with variants across the genome, affecting gene expression levels in a wide variety of tissue types
Ingested Nitrate and Breast Cancer in the Spanish Multicase-Control Study on Cancer (MCC-Spain).
BACKGROUND: Ingested nitrate leads to endogenous formation of N-nitroso compounds that are breast carcinogens in animals, but human evidence is limited. OBJECTIVE: We evaluated ingested nitrate as a risk factor for breast cancer (BC) in a multicase-control study. METHODS: Hospital-based incident BC cases and population-based controls were recruited in eight Spanish regions in 2008-2013; participants provided residential and water consumption from 18 years of age and information on known BC risk factors. Long-term nitrate levels (1940-2010) were estimated and linked with residential histories and water consumption to calculate waterborne ingested nitrate (milligrams/day). Dietary ingested nitrate (milligrams/day) was calculated using food frequency questionnaires and published dietary nitrate contents. Interactions with endogenous nitrosation factors and other variables were evaluated. A total of 1,245 cases and 1,520 controls were included in the statistical analysis. RESULTS: Among the study regions, average ± SD waterborne ingested nitrate ranged from 2.9 ± 1.9 to 13.5 ± 7.5 mg/day, and dietary ingested nitrate ranged from 88.5 ± 48.7 to 154 ± 87.8 mg/day. Waterborne ingested nitrate was not associated with BC overall, but among postmenopausal women, those with both high nitrate (> 6 vs. < 2.6 mg/day) and high red meat intake (≥ 20 vs. < 20 g/day) were more likely to be cases than women with low nitrate and low red meat intake (adjusted odds ratio = 1.64; 95% confidence interval: 1.08, 2.49; overall interaction p-value = 0.17). No association was found with dietary nitrate. CONCLUSIONS: Waterborne ingested nitrate was associated with BC only among postmenopausal women with high red meat consumption. Dietary nitrate was not associated with BC regardless of the animal or vegetable source or of menopausal status.This study was funded by the “Acción Transversal del Cáncer del Consejo de Ministros del 11/10/2007” from the “Instituto de Salud Carlos III-FEDER” (PI08/1770, PI08/0533, PI11/00226), ISCIII FIS grants. N.E.H. received financial support for the Ph.D. program from the “Agència de Gestió d’Ajuts Universitaris i de Recerca (AGAUR FI-DGR 2013) Generalitat de Cataluña.
Comparison of methods for the isolation of human breast epithelial and myoepithelial cells.
Comparison of methods for the isolation of human breast epithelial and myoepithelial cells.
Two lineages, epithelial, and myoepithelial cells are the main cell populations in the normal mammary gland and in breast cancer. Traditionally, cancer research has been performed using commercial cell lines, but primary cell cultures obtained from fresh breast tissue are a powerful tool to study more reliably new aspects of mammary gland biology, including normal and pathological conditions. Nevertheless, the methods described to date have some technical problems in terms of cell viability and yield, which hamper work with primary mammary cells. Therefore, there is a need to optimize technology for the proper isolation of epithelial and myoepithelial cells. For this reason, we compared four methods in an effort to improve the isolation and primary cell culture of different cell populations of human mammary epithelium. The samples were obtained from healthy tissue of patients who had undergone mammoplasty or mastectomy surgery. We based our approaches on previously described methods, and incorporated additional steps to ameliorate technical efficiency and increase cell survival. We determined cell growth and viability by phase-contrast images, growth curve analysis and cell yield, and identified cell-lineage specific markers by flow cytometry and immunofluorescence in 3D cell cultures. These techniques allowed us to better evaluate the functional capabilities of these two main mammary lineages, using CD227/K19 (epithelial cells) and CD10/K14 (myoepithelial cells) antigens. Our results show that slow digestion at low enzymatic concentration combined with the differential centrifugation technique is the method that best fits the main goal of the present study: protocol efficiency and cell survival yield. In summary, we propose some guidelines to establish primary mammary epithelial cell lines more efficiently and to provide us with a strong research instrument to better understand the role of different epithelial cell types in the origin of breast cancer