ABSTRACT
BACKGROUND: Studies have reported an inverse relationship between serum selenium levels and cancer incidence, but the impact of low serum selenium status on survival after a diagnosis of breast cancer has not been established. METHODS: We obtained a blood sample from 546 women diagnosed with a first primary invasive breast cancer between 2008 and 2015 in the region of Szczecin, Poland. Blood was collected after diagnosis, but prior to treatment. Serum selenium was quantified by mass spectroscopy and each patient was assigned to one of four categories (quartiles) based on the distribution in the entire cohort. Patients were followed from diagnosis to death over a mean follow-up of 3.8 years. Vital status was obtained by linkage to the Polish National Death Registry. RESULTS: The 5-year overall actuarial survival was 68.1% for women in the lowest (< 64.4 µg/L) and 82.5% for those in the highest (> 81.0 µg/L) quartile of serum selenium. In an adjusted analysis, the hazard ratio for death was 2.49 (95%CI 1.53-4.04; P = 0.0002) for patients in the lowest quartile of serum selenium, compared to those in all other quartiles. The effect of low selenium on breast cancer-specific mortality was stronger for women who were past smokers (HR 6.03; 95%CI 1.96-18.6; P = 0.0002). CONCLUSIONS: This study suggests that a selenium level in excess of 64.4 µg//L might be beneficial for women undergoing treatment for breast cancer and that selenium supplementation to achieve this level may favorably impact the outcome. Further studies are needed to confirm this association and to evaluate the impact of selenium supplementation on breast cancer survival among women with low post-diagnostic selenium levels.
Subject(s)
Breast Neoplasms/blood , Breast Neoplasms/epidemiology , Prognosis , Selenium/blood , Adult , Aged , Breast/pathology , Breast Neoplasms/pathology , Disease-Free Survival , Female , Humans , Middle Aged , Poland/epidemiology , Proportional Hazards Models , Risk FactorsABSTRACT
BACKGROUND: Thromboembolic events constitute a major health problem, despite the steadily expanding arsenal of antiplatelet drugs. Hence, there is still a need to optimize the antiplatelet therapy. OBJECTIVES: The aim of our study was to verify a hypothesis that there are no differences in platelet proteome between two groups of healthy people representing different acetylsalicylic acid (aspirin) responses as assessed by the liquid chromatography/mass spectrometry (LC/MS) technique. PATIENTS/METHODS: A total of 61 healthy volunteers were recruited for the study. Physical examination and blood collection were followed by platelet-rich plasma aggregation assays and platelet separation for proteomic LC/MS analysis. Arachidonic acid- (AA-) induced aggregation (in the presence of aspirin) allowed to divide study participants into two groups aspirin-resistant (AR) and aspirin-sensitive (AS) ones. Subsequently, platelet proteome was compared in groups using the LC/MS analysis. RESULTS: The LC/MS analysis of platelet proteome between groups revealed that out of all identified proteins, the only discriminatory protein, affecting aspirin responsiveness, is platelet carbonic anhydrase II (CA II). CONCLUSIONS: CA II is a platelet function modulator and should be taken into consideration as a cardiovascular event risk factor or therapeutic target.
Subject(s)
Aspirin/pharmacology , Blood Platelets/enzymology , Carbonic Anhydrase II/blood , Adult , Drug Resistance , Female , Humans , MaleABSTRACT
Three founder alleles of BRCA1 (C61G, 4153delA, 5382insC) were reported in Poland in 2000, and these three mutations have comprised the standard testing panel used throughout the country. However, since 2000, other recurrent mutations of BRCA1 and BRCA2 have been reported. To establish if the inclusion of one or more of these mutations will increase the sensitivity of the standard test panel, we studied 1164 Polish women with unselected breast cancer diagnosed at age of 50 or below. All women were genotyped for 12 recurrent mutations of BRCA1 and BRCA2. We identified a mutation in 83 of 1164 patients (7.1%) including 61 women with one of the original three mutations (C61G, 4153delA, 5382insC) and 22 women with a different mutation (1.9%). Three new mutations (3819del5, 185delAG and 5370C>T) were seen in multiple families. By including these three mutations in the extended panel, the mutant frequency increased from 5.2 to 6.7%. Polish women with breast cancer diagnosed at age of 50 or below should be screened with a panel of six founder mutations of BRCA1 (C61G, 4153delA, 5382insC, 3819del5, 185delAG and 5370C>T).
Subject(s)
Breast Neoplasms/epidemiology , Breast Neoplasms/genetics , Genes, BRCA1 , Genes, BRCA2 , Mutation , Adult , Age of Onset , Breast Neoplasms/diagnosis , Female , Founder Effect , Humans , Middle Aged , Ovarian Neoplasms/genetics , Poland/epidemiology , Young AdultABSTRACT
Vascular endothelial growth factor (VEGF) has been reported to mediate both osteogenesis and angiogenesis in bone regeneration. We previously found an upregulation of VEGF in adipose-derived stem cells (ASCs) when obvious mineralization occurred on a novel fluorapatite (FA)-coated surfaces. This study investigated the effect of FA and VEGF on the growth, differentiation and mineralization of (ASC) grown on ordered FA surfaces. Cells grown on FA and treated with VEGF demonstrated osteogenic differentiation as measured with ALP staining, and obvious mineralization as measured by Alizarin red staining. A combined stimulating effect of FA and VEGF was seen using both indicators. VEGF signaling pathway perturbation using a specific VEGF receptor inhibitor showed the lowest levels of ALP and Alizarin red staining, which was partially rescued when the cells were grown on FA and/or treated with the addition of VEGF. The osteogenic differentiation of ASCs stimulated by these FA surfaces as well as VEGF has been shown to be mediated through, but probably not only, the VEGF signaling pathway. The enhancement of osteogenic differentiation and mineralization supports the potential use of therapeutic VEGF and FA coatings in bone regeneration.
Subject(s)
Adipose Tissue/metabolism , Apatites/chemistry , Cell Differentiation/drug effects , Osteogenesis/drug effects , Stem Cells/metabolism , Vascular Endothelial Growth Factor A , Adipose Tissue/cytology , Adsorption , Cell Line , Humans , Stem Cells/cytology , Surface Properties , Vascular Endothelial Growth Factor A/chemistry , Vascular Endothelial Growth Factor A/pharmacologyABSTRACT
A number of genes other than BRCA1 and BRCA2 have been associated with breast cancer predisposition, and extended genetic testing panels have been proposed. It is of interest to establish the full spectrum of deleterious mutations in women with familial breast cancer.We performed whole-exome sequencing of 144 women with familial breast cancer and negative for 11 Polish founder mutations in BRCA1, CHEK2 and NBS1, and we evaluated the sequences of 12 known breast cancer susceptibility genes. A truncating mutation in a breast cancer gene was detected in 24 of 144 women (17%) with familial breast cancer. A BRCA2 mutation was detected in 12 cases, a (non-founder) BRCA1 mutation was detected in 5 cases, a PALB2 mutation was detected in 4 cases and an ATM mutation was detected in 2 cases. Polish women with familial breast cancer who are negative for founder mutations in BRCA1, CHEK2 and NBS1 should be fully screened for mutations in BRCA1, BRCA2 and PALB2. The PALB2 founder mutation c.509_519delGA should be included in the panel of Polish founder mutations.
Subject(s)
Breast Neoplasms/genetics , Genetic Predisposition to Disease , BRCA1 Protein/genetics , BRCA2 Protein/genetics , Checkpoint Kinase 2/genetics , DNA Mutational Analysis , Fanconi Anemia Complementation Group N Protein , Female , Founder Effect , Genetic Testing , Humans , Nuclear Proteins/genetics , Poland , Tumor Suppressor Proteins/geneticsABSTRACT
In previous studies, fluorapatite (FA) crystal-coated surfaces have been shown to stimulate the differentiation and mineralization of human dental pulp stem cells (DPSCs) in two-dimensional cell culture. However, whether the FA surface can recapitulate these properties in three-dimensional culture is still unknown. This study examined the differences in behavior of human DPSCs cultured on electrospun polycaprolactone (PCL) NanoECM nanofibers with or without the FA crystals. Under near-physiologic conditions, the FA crystals were synthesized on the PCL nanofiber scaffolds. The FA crystals were evenly distributed on the scaffolds. DPSCs were cultured on the PCL+FA or the PCL scaffolds for up to 28 days. Scanning electron microscope images showed that DPSCs attached well to both scaffolds after the initial seeding. However, it appeared that more multicellular aggregates formed on the PCL+FA scaffolds. After 14 days, the cell proliferation on the PCL+FA was slower than that on the PCL-only scaffolds. Interestingly, even without any induction of mineralization, from day 7, the upregulation of several pro-osteogenic molecules (dmp1, dspp, runx2, ocn, spp1, col1a1) was detected in cells seeded on the PCL+FA scaffolds. A significant increase in alkaline phosphatase activity was also seen on FA-coated scaffolds compared with the PCL-only scaffolds at days 14 and 21. At the protein level, osteocalcin expression was induced only in the DPSCs on the PCL+FA surfaces at day 21 and then significantly enhanced at day 28. A similar pattern was observed in those specimens stained with Alizarin red and Von Kossa after 21 and 28 days. These data suggest that the incorporation of FA crystals within the three-dimensional PCL nanofiber scaffolds provided a favorable extracellular matrix microenvironment for the growth, differentiation, and mineralization of human DPSCs. This FA-modified PCL nanofiber scaffold shows promising potential for future bone, dental, and orthopedic regenerative applications.
Subject(s)
Apatites/chemistry , Calcification, Physiologic/physiology , Dental Pulp/cytology , Stem Cells/physiology , Tissue Scaffolds/chemistry , Alkaline Phosphatase/analysis , Cell Adhesion/physiology , Cell Aggregation/physiology , Cell Culture Techniques , Cell Proliferation , Cellular Microenvironment , Collagen Type I/analysis , Collagen Type I, alpha 1 Chain , Core Binding Factor Alpha 1 Subunit/analysis , Crystallization , Dental Pulp/physiology , Extracellular Matrix Proteins/analysis , Humans , Materials Testing , Nanofibers/chemistry , Osteocalcin/analysis , Osteogenesis/physiology , Osteopontin/analysis , Phosphoproteins/analysis , Polyesters/chemistry , Sialoglycoproteins/analysis , Time FactorsABSTRACT
The purpose of this study is to estimate 10-year survival rates for patients with early onset breast cancer, with and without a CHEK2 mutation and to identify prognostic factors among CHEK2-positive breast cancer patients. 3,592 women with stage I to stage III breast cancer, diagnosed at or below age 50, were tested for four founder mutations in the CHEK2 gene. Information on tumor characteristics and on treatments received was retrieved from medical records. Dates of death were obtained from the Poland Vital Statistics Registry. Survival curves were generated for the mutation-positive and -negative sub-cohorts. Predictors of survival were determined among CHEK2 carriers using the Cox proportional hazards model. 3,592 patients were eligible for the study, of whom 140 (3.9 %) carried a CHEK2-truncating mutation and 347 (9.7 %) carried a missense mutation. The mean follow-up was 8.9 years. The 10-year survival for all CHEK2 mutation carriers was 78.8 % (95 % CI 74.6-83.2 %) and for non-carriers was 80.1 % (95 % CI 78.5-81.8 %). Among women with a CHEK2-positive breast cancer, the adjusted hazard ratio associated with ER-positive status was 0.88 (95 % CI 0.48-1.62). Among women with an ER-positive breast cancer, the adjusted hazard ratio associated with a CHEK2 mutation was 1.31 (95 % CI 0.97-1.77). The survival of women with breast cancer and a CHEK2 mutation is similar to that of patients without a CHEK2 mutation.
Subject(s)
Breast Neoplasms/genetics , Breast Neoplasms/mortality , Checkpoint Kinase 2/genetics , Mutation , Adult , Breast Neoplasms/enzymology , Breast Neoplasms/pathology , Female , Humans , Middle Aged , Molecular Epidemiology , Poland/epidemiology , Prognosis , Survival Rate , Young AdultABSTRACT
To identify characteristic features of breast cancers associated with an NBS1 mutation. To estimate and to compare 10-year survival rates for patients with early-onset breast cancer, with and without an NBS1 mutation. 4,566 women with stage I to stage III breast cancer, diagnosed at or below age 50, were tested for a founder mutation in the NBS1 gene. Information on tumor characteristics and on treatments received was retrieved from medical records. Dates of death were obtained from the Poland vital statistics registry. Survival curves for the mutation-positive and negative sub-cohorts were generated and were compared and the effect of an NBS1 mutation on survival was determined using the Cox proportional hazards model. 4566 patients were enrolled in the study, of whom 53 (1.2 %) carried a NBS1 mutation. Mutation carriers were similar to non-carriers in terms of tumor receptor status, grade, and lymph node status. The 10-year survival for NBS1 mutation carriers was 81.2 % (95 % CI 70.1-94.1 %) and for non-carriers was 79.4 % (95 % CI 78.0-80.9 %). The presence of an NBS1 mutation is not associated with prognosis (HR = 1.21; 95 % 0.67-2.19). The survival of women with breast cancer and a NBS1 mutation is similar to that of patients without a NBS1 mutation.
Subject(s)
Breast Neoplasms/genetics , Cell Cycle Proteins/genetics , Nuclear Proteins/genetics , Adult , Breast Neoplasms/mortality , Breast Neoplasms/pathology , Female , Founder Effect , Genetic Association Studies , Humans , Middle Aged , Prognosis , Proportional Hazards Models , Risk , Young AdultABSTRACT
BACKGROUND: Node-negative breast cancers from 2 cm to 5 cm in size are classified as stage ii, and smaller cancers, as stage i. We sought to determine if the prognosis of women with a breast cancer exactly 2 cm in size more closely resembles that of women with a stage i or a stage ii breast cancer. METHODS: Using a cohort of 4265 young women with breast cancer, we compared the 10-year breast cancer mortality rates for women who had a tumour 0.1-1.9 cm, exactly 2.0 cm, and 2.1-2.9 cm. RESULTS: In the first 3 years after diagnosis, the survival pattern of women with a 2.0-cm breast cancer was nearly identical to that of women with a larger cancer (2.1-3.0 cm). From year 3 to year 10, the relative survival of women with a 2.0-cm breast cancer was improved and nearly identical to that of women with a smaller cancer. The 10-year survival rate was 89.3% for women with tumours less than 20 mm, 86.1% for women with tumours equal to 20 mm, and 81.2% for women with 21-mm to 29-mm tumours. CONCLUSIONS: For young women with small breast cancers, the relative mortality from breast cancer is dynamic with increasing tumour size and varies with time from diagnosis.
ABSTRACT
OBJECTIVES: The injuries of the larynx constitute around 1% of all injuries. The great majority of the injuries of the larynx happens during playing. The effects of these injuries may appear insignificant however, not always the direct clinical symptoms correlate with the degree of respiratory tract failure. The symptoms of laryngeal injuries depend on the extension and strength of the trauma and always relate to impair patency of respiratory tract. The aim of the study is to compare two diagnostic methods: laryngoscopy and ultrasonography in evaluation of laryngeal injuries in children. METHODS: In the Department of Pediatric Otolaryngology, Medical University of Warsaw, in the period between 2004 and 2010 there were hospitalised 15 children with external injury of the larynx. RESULTS: From among 15 hospitalized children, 7 were qualified as not serious trauma and were treated preservatively and the other 8 as sever trauma. The mechanism of traumas was diverse. Dyspnea was a predominating symptom, the others included hoarsness, change in voice quality, even aphonia, pain while speaking and swallowing, cough and hemoptysis. CONCLUSIONS: Direct laryngoscopy is a standard in diagnostics of the injuries of the larynx. Ultrasonography of the larynx is recommended in every case of laryngeal injury as an additional non-invasive complementary diagnostic examination.
Subject(s)
Laryngeal Diseases/diagnosis , Laryngoscopy/methods , Larynx/injuries , Ultrasonography, Doppler/methods , Adolescent , Child , Child, Preschool , Cohort Studies , Dyspnea/diagnosis , Dyspnea/etiology , Female , Hospitals, University , Humans , Injury Severity Score , Laryngeal Diseases/diagnostic imaging , Male , Poland , Retrospective Studies , Sensitivity and SpecificityABSTRACT
BACKGROUND: To evaluate whether genotyping for 18 prostate cancer founder variants is helpful in identifying high-risk individuals and for determining optimal screening regimens. METHODS: A serum PSA level was measured and a digital rectal examination (DRE) was performed on 2907 unaffected men aged 40-90. Three hundred and twenty-three men with an elevated PSA (≥4 ng ml⻹) or an abnormal DRE underwent a prostate biopsy. All men were genotyped for three founder alleles in BRCA1 (5382insC, 4153delA and C61G), for four alleles in CHEK2 (1100delC, IVS2+1G>A, del5395 and I157T), for one allele in NBS1 (657del5), for one allele in HOXB13 (G84E), and for nine low-risk single-nucleotide polymorphisms (SNPs). RESULTS: On the basis of an elevated PSA or an abnormal DRE, prostate cancer was diagnosed in 135 of 2907 men (4.6%). In men with a CHEK2 missense mutation I157T, the cancer detection rate among men with an elevated PSA or an abnormal DRE was much higher (10.2%, P=0.0008). The cancer detection rate rose with the number of SNP risk genotypes observed from 1.2% for men with no variant to 8.6% for men who carried six or more variants (P=0.04). No single variant was helpful on its own in predicting the presence of prostate cancer, however, the combination of all rare mutations and SNPs improved predictive power (area under the curve=0.59; P=0.03). CONCLUSION: These results suggest that testing for germline CHEK2 mutations improves the ability to predict the presence of prostate cancer in screened men, however, the clinical utility of incorporating DNA variants in the screening process is marginal.
Subject(s)
Early Detection of Cancer/methods , Founder Effect , Genotyping Techniques , Germ-Line Mutation , Prostatic Neoplasms/diagnosis , Protein Serine-Threonine Kinases/genetics , Adult , Aged , Aged, 80 and over , Alleles , Checkpoint Kinase 2 , Genetic Predisposition to Disease/genetics , Genotype , Humans , Male , Mass Screening/methods , Middle Aged , Precision Medicine/methods , Prostatic Neoplasms/genetics , Risk FactorsABSTRACT
AIM: To synthesise fluoridated hydroxyapatite (FA) crystals directly on preformed metal crowns (PMCs) and evaluate the anti-cariogenic properties in an in vitro model. METHODS: FA crystals were grown on etched PMCs and stainless steel discs and characterised by SEM. FA-coated discs allowed fluoride release to be assessed from a known surface area of FA crystals. Discs were divided into four groups (n = 6/group) and exposed to solutions at pH 4-7. Fluoride levels in solution were measured after each exposure. Twelve FA-coated and 12 non-coated PMCs were cemented onto human molars using glass ionomer (GI) or unfilled resin, making four groups of six teeth; FA-coated + GI, FA-coated + resin; non-coated + GI and non-coated + resin. Teeth were exposed to acidified gelatin (pH = 4.3) for 9 weeks. RESULTS: SEM showed FA crystal growth on interior and exterior of the crowns. Average fluoride release from FA-coated discs was 0.16 mg/L/cm² at pH < 5.0. Teeth were sectioned through the lesion. Polarised microscopic examination revealed significantly smaller lesions in FA-coated crown groups compared to non-coated crown groups. CONCLUSION: FA-coated PMCs demonstrated carious lesion preventing effects, i.e. fluoride release and reduction of demineralisation at crown/tooth interface. FA-coated crowns could be an aesthetic, inexpensive and caries preventive alternative in clinical dentistry.
Subject(s)
Dental Caries , Glass Ionomer Cements , Crowns , Dental Caries/prevention & control , Glass Ionomer Cements/chemistry , Humans , Molar , Stainless Steel/chemistryABSTRACT
BACKGROUND: To establish the contribution of eight founder alleles in three DNA damage repair genes (BRCA1, CHEK2 and NBS1) to prostate cancer in Poland, and to measure the impact of these variants on survival among patients. METHODS: Three thousand seven hundred fifty men with prostate cancer and 3956 cancer-free controls were genotyped for three founder alleles in BRCA1 (5382insC, 4153delA, C61G), four alleles in CHEK2 (1100delC, IVS2+1G>A, del5395, I157T), and one allele in NBS1 (657del5). RESULTS: The NBS1 mutation was detected in 53 of 3750 unselected cases compared with 23 of 3956 (0.6%) controls (odds ratio (OR)=2.5; P=0.0003). A CHEK2 mutation was seen in 383 (10.2%) unselected cases and in 228 (5.8%) controls (OR=1.9; P<0.0001). Mutation of BRCA1 (three mutations combined) was not associated with the risk of prostate cancer (OR=0.9; P=0.8). In a subgroup analysis, the 4153delA mutation was associated with early-onset (age ≤ 60 years) prostate cancer (OR=20.3, P=0.004). The mean follow-up was 54 months. Mortality was significantly worse for carriers of a NBS1 mutation than for non-carriers (HR=1.85; P=0.008). The 5-year survival for men with an NBS1 mutation was 49%, compared with 72% for mutation-negative cases. CONCLUSION: A mutation in NBS1 predisposes to aggressive prostate cancer. These data are relevant to the prospect of adapting personalised medicine to prostate cancer prevention and treatment.
Subject(s)
Cell Cycle Proteins/genetics , Nuclear Proteins/genetics , Prostatic Neoplasms/genetics , Prostatic Neoplasms/mortality , Adult , Aged , Aged, 80 and over , BRCA1 Protein/genetics , Biomarkers, Tumor/genetics , Checkpoint Kinase 2 , Genes, BRCA1 , Genetic Predisposition to Disease , Genotype , Humans , Kaplan-Meier Estimate , Male , Middle Aged , Mutation , Prognosis , Protein Serine-Threonine Kinases/geneticsABSTRACT
BACKGROUND: The variable penetrance of breast cancer in BRCA1/2 mutation carriers suggests that other genetic or environmental factors modify breast cancer risk. Two genes of special interest are prohibitin (PHB) and methylene-tetrahydrofolate reductase (MTHFR), both of which are important either directly or indirectly in maintaining genomic integrity. METHODS: To evaluate the potential role of genetic variants within PHB and MTHFR in breast and ovarian cancer risk, 4102 BRCA1 and 2093 BRCA2 mutation carriers, and 6211 BRCA1 and 2902 BRCA2 carriers from the Consortium of Investigators of Modifiers of BRCA1 and BRCA2 (CIMBA) were genotyped for the PHB 1630 C>T (rs6917) polymorphism and the MTHFR 677 C>T (rs1801133) polymorphism, respectively. RESULTS: There was no evidence of association between the PHB 1630 C>T and MTHFR 677 C>T polymorphisms with either disease for BRCA1 or BRCA2 mutation carriers when breast and ovarian cancer associations were evaluated separately. Analysis that evaluated associations for breast and ovarian cancer simultaneously showed some evidence that BRCA1 mutation carriers who had the rare homozygote genotype (TT) of the PHB 1630 C>T polymorphism were at increased risk of both breast and ovarian cancer (HR 1.50, 95%CI 1.10-2.04 and HR 2.16, 95%CI 1.24-3.76, respectively). However, there was no evidence of association under a multiplicative model for the effect of each minor allele. CONCLUSION: The PHB 1630TT genotype may modify breast and ovarian cancer risks in BRCA1 mutation carriers. This association need to be evaluated in larger series of BRCA1 mutation carriers.
Subject(s)
Breast Neoplasms/genetics , Genes, BRCA1 , Genes, BRCA2 , Methylenetetrahydrofolate Reductase (NADPH2)/genetics , Ovarian Neoplasms/genetics , Polymorphism, Genetic , Repressor Proteins/genetics , Female , Genetic Predisposition to Disease , Heterozygote , Humans , Mutation , Prohibitins , RiskABSTRACT
UNLABELLED: To study how apatite crystal alignment of an enamel-like substrate affects DPSC cellular adhesion and growth as a precursor to produce an in vitro enamel/dentin superstructure for future studies. The cells were subcultured in 10% FBS DMEM up to seven weeks on the two surfaces. Specimens were observed under SEM, counted, and analyzed using the human pathway-focused matrix and adhesion PCR array. After three days, the cell number on ordered FA surface was significantly higher than on the disordered surface. Of the 84 focused pathway genes, a total of 20 genes were either up or down regulated in the cells on ordered FA surface compared to the disordered surface. More interestingly, of the cell-matrix adhesion molecules, integrin alpha 7 and 8 (ITGA 7 and 8), integrin beta 3 and 4 (ITGB3 and 4), and the vitronectin receptor-integrin alpha V (ITGAV) and the key adhesion protein-fibronectin1 (FN1) were up-regulated. In SEM, both surfaces showed good biocompatibility and supported long term growth of DPSC cells but with functional cell-matrix interaction on the ordered FA surfaces. SIGNIFICANCE: The enhanced cellular response of DPSC cell to the ordered FA crystal surface involves a set of delicately regulated matrix and adhesion molecules which could be manipulated by treating the cells with a dentin extract, to produce a dentin/enamel superstructure.
Subject(s)
Apatites/pharmacology , Dental Enamel/chemistry , Dental Pulp/cytology , Stem Cells/cytology , Stem Cells/drug effects , Apatites/chemical synthesis , Cell Adhesion/drug effects , Cell Count , Cell Proliferation/drug effects , Crystallization , Extracellular Matrix Proteins/genetics , Extracellular Matrix Proteins/metabolism , Gene Expression Regulation/drug effects , Humans , Polymerase Chain Reaction , Stem Cells/metabolism , Stem Cells/ultrastructure , Surface Properties/drug effectsABSTRACT
We have studied brain stem cells in the ewe brain that project to the bed nucleus of the stria terminalis (BNST) and determined if these cells are activated by estradiol-17beta. This would predicate an indirect role in the estradiol-17beta regulation of gonadotropin releasing hormone (GnRH) cells, since these receive input from the BNST. Ovariectomized ewes received 50 mug estradiol-17beta benzoate (i.m.) 1 h prior to brain collection, so that activated cells could be identified by Fos immunohistochemistry. Retrograde tracer (FluoroGold; FG), was injected into the three divisions of the BNST and labeled cells were mapped to the A1 and A2 regions and the parabrachial nucleus (PBN) of the brain stem. With FG injection into the dorsal and lateral BNST, all FG-containing cells in the caudal A1 and 45% of those in A2 stained for dopamine-beta-hydroxylase (DBH), indicating noradrenergic type. No FG-labelled cells in the PBN were DBH-positive. In A1 and A2 respectively, 42% and 46% of FG-labelled cells were Fos-positive, with no double-labeling in cells of the PBN. In ewes receiving FG injections into the ventral BNST, estrogen receptor (ER)alpha-immunoreactive nuclei were found in 82% of A1-FG labeled and 38% of A2-FG labeled cells. No FG-labelled cells of the PBN were ERalpha-positive. Anterograde tracing from A1 with microruby injection identified projections to the PBN, BNST and preoptic area (POA). Thus, A1 and A2 noradrenergic neurons project to the BNST in the ewe brain, express ERalpha and are activated by estradiol-17beta. These noradrenergic, estrogen-responsive cells may provide indirect input to GnRH cells, via the BNST.
Subject(s)
Brain Stem/drug effects , Estradiol/analogs & derivatives , Estrogens/pharmacology , Receptors, Adrenergic/metabolism , Septal Nuclei/drug effects , Animals , Brain Stem/metabolism , Cell Nucleus/drug effects , Cell Nucleus/metabolism , Dopamine beta-Hydroxylase/metabolism , Estradiol/pharmacology , Estrogen Receptor alpha/metabolism , Female , Gonadotropin-Releasing Hormone/metabolism , Immunohistochemistry , Neural Pathways/drug effects , Neural Pathways/metabolism , Neuronal Tract-Tracers , Ovariectomy , Preoptic Area/drug effects , Preoptic Area/metabolism , Proto-Oncogene Proteins c-fos/metabolism , Septal Nuclei/metabolism , SheepABSTRACT
In the Milan area (Northern Italy), we identified a family characterized by a high prevalence of ovarian and breast cancer cases (5 out of 6 subjects, over 3 generations), and a predominant prevalence of ovarian lesions (4 out of 5 patients). Analysis of BRCA1 and BRCA2 genes allowed the identification of the missense c.190T>C mutation in codon 64 (Cys64Arg) of BRCA1. The aims of the present investigation were to characterize the functional implications of the c.190T>C mutation at the molecular level, and to search whether additional polymorphisms might be linked to the peculiar phenotypic features observed in the Italian pedigree. Molecular modelling studies suggested that substitution of the cysteine 64 with an arginine likely disrupts the architecture of the BRCA1 RING finger domain, responsible for the interaction with BARD1, essential for the tumor-suppressor activity of the BRCA1-BARD1 complex. By splicing site information analysis, exonic splicing enhancer site characterization, and analysis of transcript fragment length and sequence, we showed that the c.190T>C mutation was able to modulate the splicing of exon 5 in a fashion opposite to the c.190T>G transversion, responsible for the functionally-related Cys64Gly amino acid substitution. Genotyping of BRCA1 and BRCA2 in the Italian family revealed the presence of two significant polymorphisms: the cancer-associated c.2612C>T SNP in BRCA1, and the c.-26G>A SNP in the BRCA2 gene, acting as an ovarian cancer risk modifier in carriers of deleterious BRCA1 mutations. Analysis of these SNPs in a genotypically-unrelated Polish family, characterized by prevalent breast neoplasms in carriers of the c.190T>C mutation, revealed a genetic profile consistent with the hypothetic role of both polymorphisms.
Subject(s)
BRCA1 Protein/genetics , BRCA1 Protein/physiology , Mutation, Missense , Amino Acid Sequence , Apoptosis Regulatory Proteins , BRCA2 Protein/genetics , BRCA2 Protein/physiology , Base Sequence , Codon , Exons , Female , Genes, BRCA1 , Humans , Male , Molecular Sequence Data , Ovarian Neoplasms/genetics , Ovarian Neoplasms/metabolism , Pedigree , Polymorphism, GeneticABSTRACT
BACKGROUND: Carriers of heterozygous mutations in CHEK2 or BRCA1 are at increased risk of breast cancer. These mutations are rare and a very small number of women in a population will carry two mutations. However, it is of interest to estimate the breast cancer risks associated with carrying two mutations because this information may be informative for genetic counsellors and may provide clues to the carcinogenic process. METHODS: We genotyped 7782 Polish breast cancer patients and 6233 controls for seven founder mutations in BRCA1 and CHEK2. Odds ratios (OR) and 95% confidence intervals (CI) were estimated for the mutations, singly and in combination. RESULTS: Of the 7782 women with breast cancer, 1091 had one mutation (14.0%) and 37 had two mutations (0.5%). Compared to controls, the odds ratio for a BRCA1 mutation in isolation was 13.1 (95% CI 8.2 to 21). The odds ratio was smaller for BRCA1 mutation carriers who also carried a CHEK2 mutation (OR 6.6, 95% CI 1.5 to 29), but the difference was not statistically significant. In contrast, the odds ratio for women who carried two CHEK2 mutations (OR 3.9, 95% CI 1.5 to 10) was greater than that for women who carried one CHEK2 mutation (OR 1.9, 95% CI 1.6 to 2.1). The odds ratio for women who carried both a truncating mutation and the missense mutation in CHEK2 was 7.0 (95% CI 0.9 to 56) and was greater than for women who carried the truncating mutation alone (OR 3.3, 95% CI 2.4 to 4.3) or the missense mutation alone (OR 1.6, 95% CI 1.4 to 1.9), but the difference was not statistically significant. CONCLUSION: Our study suggests that the risk of breast cancer in carriers of a deleterious CHEK2 mutation is increased if the second allele is the I157T missense variant. However, the presence of a CHEK2 mutation in women with a BRCA1 mutation may not increase their risk beyond that of the BRCA1 mutation alone. These suggestive findings need to be verified in other studies.
Subject(s)
BRCA1 Protein/genetics , Breast Neoplasms/genetics , Mutation, Missense , Protein Serine-Threonine Kinases/genetics , Adolescent , Adult , Aged , Aged, 80 and over , Checkpoint Kinase 2 , Female , Genetic Predisposition to Disease , Genetic Variation , Genotype , Humans , Middle Aged , Odds Ratio , Prospective Studies , Risk FactorsABSTRACT
To investigate the relationship between CHEK2 mutation status and estrogen receptor (ER) status in unselected cases of early-onset breast cancer from Poland, we screened 4441 women diagnosed with breast cancer younger than 51 years and 7217 controls for three inherited mutations in CHEK2 (1100delC, IVS2+1G>A, del5395). ER status was compared between CHEK2-positive and CHEK2-negative breast cancer cases. A truncating mutation in CHEK2 was seen in 140 of 4441 cases and in 70 of 7217 controls [odds ratio (OR) = 3.3; 95% CI = 2.5-4.4; p < 0.0001]. ER status was available for 92 of 140 mutation carriers and for 3001 of 4301 non-carriers with breast cancer. The OR was higher for ER-positive cancers (OR = 3.9; 95% CI = 2.7-5.4; p < 0.0001) than for ER-negative cancers (OR = 2.1; 95% CI = 1.3-3.3; p = 0.002). Sixty-six of the 92 breast cancers in carriers of CHEK2 truncating mutations were ER positive compared with 1742 of the 3001 breast cancers in non-carriers (72% vs 58%; p = 0.01). Women with a CHEK2 mutation face a fourfold increase in the risk of ER-positive breast cancer and might be candidates for tamoxifen chemoprevention.