ABSTRACT
Primary hyperparathyroidism (PHPT) is associated with hypovitaminosis D as assessed by serum total 25-hydroxyvitamin D (TotalD) levels. The aim of this study is to evaluate whether this is also the case for the calculated bioavailable 25-hydroxyvitamin D (BioD) or free 25-hydroxyvitamin D (FreeD), and whether the vitamin D status is influenced by genetic background. We compared vitamin D status of 88 PHPT patients each with a matched healthy family member sharing genetic background, i.e., first-degree relative (FDR), or not, namely an in-law relative (ILR). We compared TotalD and vitamin D-binding protein (DBP), using the latter to calculate BioD and FreeD. We also genotyped two common DBP polymorphisms (rs7041 and rs4588) likely to affect the affinity for and levels of vitamin D metabolites. TotalD was lower (p < 0.001) in PHPT (12.3 ± 6.6 ng/mL) than either family member group (FDR: 19.4 ± 12.1 and ILR: 23.2 ± 14.1), whether adjusted for DBP or not. DBP levels were also significantly lower (p < 0.001) in PHPT (323 ± 73 mg/L) versus FDR (377 ± 98) or ILR (382 ± 101). The differences between PHPT and control groups for TotalD, BioD, and FreeD were maintained after adjustment for season, gender, and serum creatinine. 25-hydroxyvitamin D, evaluated as total, free, or bioavailable fractions, is decreased in PHPT. No difference was seen between first-degree relative and in-law controls, suggesting that neither genetic nor non-genetic background greatly influences the genesis of the hypovitaminosis D seen in PHPT.
Subject(s)
Family , Hyperparathyroidism, Primary/genetics , Polymorphism, Single Nucleotide , Vitamin D Deficiency/genetics , Vitamin D-Binding Protein/genetics , Vitamin D/analogs & derivatives , Adult , Aged , Female , Humans , Hyperparathyroidism, Primary/blood , Hyperparathyroidism, Primary/complications , Male , Middle Aged , Seasons , Sex Factors , Vitamin D/blood , Vitamin D Deficiency/blood , Vitamin D Deficiency/complicationsABSTRACT
BACKGROUND: Recent studies investigating the single cytochrome P450 (CYP) 2D6 allele *2A reported an association with the response to drug treatments. More genetic data can be obtained, however, by high-throughput based-technologies. Aim of this study is the high-throughput analysis of the CYP2D6 polymorphisms to evaluate its effectiveness in the identification of patient responders/non-responders to CYP2D6-metabolized drugs. METHODS: An attempt to compare our results with those previously obtained with the standard analysis of CYP2D6 allele *2A was also made. Sixty blood samples from patients treated with CYP2D6-metabolized drugs previously genotyped for the allele CYP2D6*2A, were analyzed for the CYP2D6 polymorphisms with the AutoGenomics INFINITI CYP4502D6-I assay on the AutoGenomics INFINITI analyzer. RESULTS: A higher frequency of mutated alleles in responder than in non-responder patients (75.38 % vs 43.48 %; p = 0.015) was observed. Thus, the presence of a mutated allele of CYP2D6 was associated with a response to CYP2D6-metabolized drugs (OR = 4.044 (1.348 - 12.154). No difference was observed in the distribution of allele *2A (p = 0.320). CONCLUSIONS: The high-throughput genetic analysis of the CYP2D6 polymorphisms better discriminate responders/non-responders with respect to the standard analysis of the CYP2D6 allele *2A. A high-throughput genetic assay of the CYP2D6 may be useful to identify patients with different clinical responses to CYP2D6-metabolized drugs.
Subject(s)
Cytochrome P-450 CYP2D6/genetics , High-Throughput Nucleotide Sequencing , Polymorphism, Genetic , Alleles , Alzheimer Disease/drug therapy , Alzheimer Disease/genetics , Biotransformation/genetics , Cytochrome P-450 CYP2D6/deficiency , Cytochrome P-450 CYP2D6/metabolism , Donepezil , Drug Resistance/genetics , Gene Deletion , Gene Duplication , Gene Frequency , Genotype , Humans , Indans/pharmacokinetics , Indans/therapeutic use , Nootropic Agents/pharmacokinetics , Nootropic Agents/therapeutic use , Piperidines/pharmacokinetics , Piperidines/therapeutic use , Polymorphism, Single Nucleotide , Sampling Studies , Single-Blind MethodABSTRACT
BACKGROUND: The development of non-invasive procedure to determine HER2 status may represent a powerful method for monitoring disease progression and response to the treatment. METHODS: Serum samples and RNA from peripheral blood were evaluated in 85 breast cancer patients (49 HER2 positive and 36 HER2 negative) and 22 healthy controls. HER2 mRNA levels were measured by real-time quantitative PCR (QPCR) and serum HER2 protein by immunoenzimatic assay (EIA). ROC curve analyses were used to determine the optimal cut off values. RESULTS: A statistically significant difference was detected for both QPCR and EIA in HER2 positive patients as compared with both healthy controls and HER2 negative tumours. QPCR showed a 91% (CI95%: 84%-98%) specificity and a 78% (CI95%: 68%-88%) sensitivity for an optimal cut off value of 4.74. The optimal cut off value for EIA was 22 ng/ml yielding a 95% (CI95%: 90%-100%) specificity and a 59% (CI95%: 48%-70%) sensitivity. The QPCR assay was slightly less specific than EIA in discriminating HER2 positive breast cancers from HER2 negative tumours (78% CI95%: 69%-87% versus 86% CI95%: 79%-93%), but it was more sensitive (76% CI95%: 67%-85% versus 55% CI95%: 44%-66%). CONCLUSIONS: Our results indicate that QPCR performs better than EIA in the determination of HER2 status of breast cancer patients and could be useful in monitoring the disease during follow up.
Subject(s)
Breast Neoplasms/blood , Breast Neoplasms/genetics , Enzyme-Linked Immunosorbent Assay/methods , Gene Dosage , Polymerase Chain Reaction/methods , Receptor, ErbB-2/blood , Receptor, ErbB-2/genetics , Adult , Breast Neoplasms/metabolism , Breast Neoplasms/pathology , Case-Control Studies , Disease Progression , Female , Humans , Middle Aged , RNA, Messenger/blood , RNA, Messenger/genetics , Receptor, ErbB-2/metabolismABSTRACT
BACKGROUND: Amplification of the Her2neu oncogene is a well known indicator of poor prognosis in breast cancer patients. The implementation into the clinical setting of therapeutical strategies directly targeting the Her2neu gene product, has create the need for the development of non-invasive analytical techniques in order to monitoring minimal residual disease and response to the treatment. METHODS: To detect the expression of Her2neu mRNA in peripheral blood, we developed a specific real-time quantitative reverse transcription-PCR (QPCR) assay. The analyses were performed on blood samples obtained from 30 breast cancer patients positive for Her2neu overexpression by immunohistochemical analysis (IHC), 10 breast cancer patients negative for Her2neu overexpression, and 24 healthy controls. RESULTS: Her2neu positive tumors showed a significant increase in mRNA transcripts as compared with both healthy controls (n=24) and Her2neu negative patients (n=10). After establishing a cut-off value, 18 out of the 30 Her2neu positive patient scored positive for Her2neu expression, whereas only 1 out of the 10 Her2neu negative patients was weakly positive. CONCLUSIONS: Her2neu QPCR is suitable method for Her2neu overexpression detection in peripheral blood from clinical samples of breast cancer patients. QPCR could be used to identify breast cancer patients with poor prognosis and for monitoring response to the therapy.
