Jorge Villanova Biazús

Ultra-high performance liquid chromatography tandem mass spectrometric method for the determination of tamoxifen, N-desmethyltamoxifen, 4-hydroxytamoxifen and endoxifen in dried blood spots--development, validation and clinical application during breast cancer adjuvant therapy.

A LC-MSMS method for the simultaneous determination of tamoxifen, N-desmethyltamoxifen, 4-hydroxytamoxifen and endoxifen in dried blood spots samples was developed and validated. The method employs an ultrasound-assisted liquid extraction and a reversed phase separation in an Acquity(®) C18 column (150×2.1 mm, 1.7 µm). Mobile phase was a mixture of formic acid 0.1% (v/v) pH 2.7 and acetonitrile (gradient from 60:40 to 50:50, v/v). Total analytical run time was 8 min. Precision assays showed CV % lower than 10.75% and accuracy in the range 94.5 to 110.3%. Mean analytes recoveries from DBS ranged from 40% to 92%. The method was successfully applied to 91 paired clinical DBS and plasma samples. Dried blood spots concentrations were highly correlated to plasma, with rs>0.83 (P<0.01). Median estimated plasma concentrations after hematocrit and partition factor adjustment were: TAM 123.3 ng mL(-1); NDT 267.9 ng mL(-1), EDF 10.0 ng mL(-1) and HTF 1.3 ng mL(-1,) representing in average 98 to 104% of the actually measured concentrations. The DBS method was able to identify 96% of patients with plasma EDF concentrations below the clinical threshold related to better prognosis (5.9 ng mL(-1)). The procedure has adequate analytical performance and can be an efficient tool to optimize adjuvant breast cancer treatment, especially in resource limited settings.

Sexual function and chemotherapy in postmenopausal women with breast cancer

BackgroundThis cross-sectional, nested cohort study assessed Female Sexual Function Index (FSFI) scores in postmenopausal women with breast cancer receiving primary chemotherapy.MethodsThe FSFI questionnaire was administered to 24 postmenopausal women one month after diagnosis of breast cancer (post-diagnosis group) and one month after completion of the first cycle of primary anthracyclin-based chemotherapy (post-chemotherapy group). Scores were compared to those of 24 healthy postmenopausal women seeking routine gynecological care (control group). All patients were sexually active at the time of enrollment. Mean age was 57.29 ± 11.82 years in the breast cancer group and 52.58 ± 7.19 years in the control group.ResultsScores in all domains of the FSFI instrument were significantly lower in the post-diagnosis group than in controls (−41.3%, p < 0.001). A further major reduction in FSFI scores was evident on completion of one cycle of primary chemotherapy (down 46.7% from post-diagnosis scores, p < 0.003), again in all domains. Six patients (25%) ceased all sexual relations, in a significant change from baseline (p < 0.001). After one chemotherapy cycle, a further five patients ceased sexual activity, for a total of 11 (45.8%) participants – a borderline significant difference (p = 0.063).ConclusionThe present study shows that female sexual function as assessed by the FSFI declines significantly at two distinct points in time: upon diagnosis of breast cancer and after administration of systemic chemotherapy.

Reproducibility of three classification systems of ductal carcinoma in situ of the breast using a web-based survey

This study assessed the degree of diagnostic agreement among pathologists between three classification systems of ductal carcinoma in situ of the breast (DCIS). Thirteen pathologists received the same set of digitized images of microscopy of 43 DCIS cases and answered a questionnaire containing the criteria to compose the three classification systems studied: Holland, modified Lagios, and Van Nuys. A computer program was created, which organizes the information collected from each pathologist, supplying the histological grading of the cases within the three classification systems. The results were analyzed using percental agreement and the Kappa test. Diagnostic agreement for the three DCIS of the breast classification systems presented K values that varied from 0.27 to 0.37. Among the three classifications used, most agreement was for Van Nuys, showing a Kappa index of 0.37. These results matched the interobserver agreements, with Kappa indices varying from 0.13 to 0.64 for the Holland classification; 0.23 to 0.61 for the modified Lagios classification; and 0.23 to 0.74 for the Van Nuys classification. Pathologists specialized in breast pathology showed greater reproducibility for all the criteria evaluated. Comparing the three classification systems, diagnostic agreement and accuracy were rated higher for the classification of Van Nuys compared to modified Lagios and Holland.

Analysis of KIR gene frequencies and HLA class I genotypes in breast cancer and control group.

Breast cancer is the main cause of cancer-related death among women, with a 0.5% increase in incidence per year. Natural killer cells (NK) are part of the innate immune system recognizing class I HLA molecules on target cells through their membrane receptors, called killer cell immunoglobulin-like receptors (KIR). The aim of our study was to evaluate the association between the KIR genes and HLA alleles in patients with breast cancer and healthy controls. Two hundred thirty patients with breast cancer and 272 healthy controls were typed for HLA class I and KIR genes by PCR-SSO. When both groups were compared, the presence of inhibitory KIR2DL2 receptors was significantly higher in breast cancer patients than in healthy controls. No significant differences were found for HLA-C2 and HLA-Bw4. However, a higher frequency of HLA-C1 in breast cancer patients was observed. These findings suggest a potential role for the KIR gene system in breast cancer. Further studies to confirm this observation are warranted.

CYP3A4*22 is related to increased plasma levels of 4-hydroxytamoxifen and partially compensates for reduced CYP2D6 activation of tamoxifen.

AIM To evaluate the impact of CYP3A4*22 in the formation of endoxifen (EDF) and hydroxytamoxifen (HTF), under different CYP2D6 genotypic backgrounds. MATERIALS & METHODS 178 patients were enrolled in the study. CYP2D6 and CYP3A4 genotyping and tamoxifen (TAM) and metabolites quantification were performed. RESULTS EDF concentrations were lower in poor (2.77 ng ml(-1)) and CYP2D6 intermediate metabolizers (5.84 ng ml(-1)), comparing to functional group (EM-F) (10.67 ng ml(-1), p < 0.001). HTF and TAM levels were respectively 47 and 53% higher in CYP3A4*22 carriers compared with *1/*1 patients in the whole group. Patients with impaired CYP2D6 metabolism and carriers of CYP3A4*22 had EDF levels comparable to CYP2D6 EM-F group (9.06 and 10.67 ng ml(-1), p = 0.247). CONCLUSION The presence of CYP3A4*22 might compensate the reduction of EDF concentrations related to CYP2D6 inactivity, especially due to increased HTF concentrations.

Development, validation and clinical application of a HPLC-FL method for CYP2D6 phenotyping in South Brazilian breast cancer patients

OBJECTIVE To develop and validate a method for determination of dextromethorphan (DMT) and dextrorphan (DTP) in plasma samples using HPLC-FL and to apply it to CYP2D6 phenotyping of a population from the South of Brazil. METHODS Samples were prepared by hydrolysis and liquid-liquid extraction. Analysis was conducted in a reversed phase column, with isocratic elution and fluorescence detection. One hundred and forty patients being treated with tamoxifen were given 30 mg of dextromethorphan and their CYP2D6 phenotypes were determined on the basis of [DMT]/[DTP] metabolic ratios in plasma samples collected after 3h. RESULTS Total chromatography running time was 12 min. Precision (CV%) was below 9.7% and accuracy was between 92.1 and 106.9%. The lower limits of quantification were 1 ng mL(-1) for DMT and 10 ng mL(-1) for DTP. Mean extraction yield of analytes was 86.6%. Mean age of patients was 55.7 years. Phenotype frequencies were as follows: 7.1% poor metabolizers, 13.6% intermediate metabolizers, 77.1% extensive metabolizers and 2.1 ultra-rapid metabolizers. Metabolic ratios for patients on strong (n=11) and weak (n=16) CYP2D6 activity inhibitors were different from each other and also different from ratios for patients not taking enzyme inhibitors (n=113). CONCLUSIONS A sensitive method for determination of dextromethorphan and its metabolite in plasma samples was developed and successfully applied, providing evidence of the impact that CYP2D6 inhibitors have on the enzymes metabolic capacity.

Immediate reconstruction with autologous fat grafting: influence in loco regional recurrence in breast cancer

Objective to evaluate local and systemic recurrence of breast cancer in patients submitted to autologous fat grafting in the immediate reconstruction after conservative surgery for breast cancer. Methods this is a historical cohort study comparing 167 patients submitted to conservative surgery without reconstruction (conservative surgery group) with 27 patients submitted to conservative treatment with immediate graft reconstruction, following the Colemans technique (lipofilling group). All patients had invasive carcinoma and were operated by a single surgeon from 2004 to 2011. The postoperative follow-up time was 36 months. Results the overall incidence of local recurrence was 2.4%. No patient in the lipofilling group had local recurrence during the study period. For systemic recurrence, the rates obtained were 3.7% (one patient) for the fat grafting group and 1.8% (three patients) for the conservative surgery group without reconstruction. Conclusion there was no significant difference for local or systemic recurrence in the groups studied. Immediate autologous fat grafting appears to be a safe procedure..................................................................................................... 14 INTRODUÇÃO ................................................................................................ 16 REVISÃO DA LITERATURA ........................................................................... 19 1 Estratégias para localizar e selecionar as informações ............................ 19 1.1 Marco conceitual esquemático .......................................................... 20 2 Cirurgia conservadora .............................................................................. 20 3 Enxerto autólogo de gordura .................................................................... 25 3.1 Lipoenxertia ....................................................................................... 25 3.2 Biologia .............................................................................................. 25 3.3 Técnica .............................................................................................. 27 3.4 Conceito de enxerto estruturado de gordura de Coleman ................. 29 3.5 Indicações ......................................................................................... 33 3.6 Risco oncológico................................................................................ 34 3.7 Seguimento Radiologico .................................................................... 40 JUSTIFICATIVA .............................................................................................. 42 HIPÓTESES ................................................................................................... 43 OBJETIVOS .................................................................................................... 44 REFERÊNCIAS .............................................................................................. 45 ARTIGO EM INGLÊS...................................................................................... 53

Immediate reconstruction with autologous fat grafting: influence in breast cancerregional recurrence

Objective to evaluate local and systemic recurrence of breast cancer in patients submitted to autologous fat grafting in the immediate reconstruction after conservative surgery for breast cancer. Methods this is a historical cohort study comparing 167 patients submitted to conservative surgery without reconstruction (conservative surgery group) with 27 patients submitted to conservative treatment with immediate graft reconstruction, following the Colemans technique (lipofilling group). All patients had invasive carcinoma and were operated by a single surgeon from 2004 to 2011. The postoperative follow-up time was 36 months. Results the overall incidence of local recurrence was 2.4%. No patient in the lipofilling group had local recurrence during the study period. For systemic recurrence, the rates obtained were 3.7% (one patient) for the fat grafting group and 1.8% (three patients) for the conservative surgery group without reconstruction. Conclusion there was no significant difference for local or systemic recurrence in the groups studied. Immediate autologous fat grafting appears to be a safe procedure..................................................................................................... 14 INTRODUÇÃO ................................................................................................ 16 REVISÃO DA LITERATURA ........................................................................... 19 1 Estratégias para localizar e selecionar as informações ............................ 19 1.1 Marco conceitual esquemático .......................................................... 20 2 Cirurgia conservadora .............................................................................. 20 3 Enxerto autólogo de gordura .................................................................... 25 3.1 Lipoenxertia ....................................................................................... 25 3.2 Biologia .............................................................................................. 25 3.3 Técnica .............................................................................................. 27 3.4 Conceito de enxerto estruturado de gordura de Coleman ................. 29 3.5 Indicações ......................................................................................... 33 3.6 Risco oncológico................................................................................ 34 3.7 Seguimento Radiologico .................................................................... 40 JUSTIFICATIVA .............................................................................................. 42 HIPÓTESES ................................................................................................... 43 OBJETIVOS .................................................................................................... 44 REFERÊNCIAS .............................................................................................. 45 ARTIGO EM INGLÊS...................................................................................... 53

Reconstrução imediata com enxerto autólogo de gordura : influência na recorrência local de câncer de mama

Objective to evaluate local and systemic recurrence of breast cancer in patients submitted to autologous fat grafting in the immediate reconstruction after conservative surgery for breast cancer. Methods this is a historical cohort study comparing 167 patients submitted to conservative surgery without reconstruction (conservative surgery group) with 27 patients submitted to conservative treatment with immediate graft reconstruction, following the Colemans technique (lipofilling group). All patients had invasive carcinoma and were operated by a single surgeon from 2004 to 2011. The postoperative follow-up time was 36 months. Results the overall incidence of local recurrence was 2.4%. No patient in the lipofilling group had local recurrence during the study period. For systemic recurrence, the rates obtained were 3.7% (one patient) for the fat grafting group and 1.8% (three patients) for the conservative surgery group without reconstruction. Conclusion there was no significant difference for local or systemic recurrence in the groups studied. Immediate autologous fat grafting appears to be a safe procedure..................................................................................................... 14 INTRODUÇÃO ................................................................................................ 16 REVISÃO DA LITERATURA ........................................................................... 19 1 Estratégias para localizar e selecionar as informações ............................ 19 1.1 Marco conceitual esquemático .......................................................... 20 2 Cirurgia conservadora .............................................................................. 20 3 Enxerto autólogo de gordura .................................................................... 25 3.1 Lipoenxertia ....................................................................................... 25 3.2 Biologia .............................................................................................. 25 3.3 Técnica .............................................................................................. 27 3.4 Conceito de enxerto estruturado de gordura de Coleman ................. 29 3.5 Indicações ......................................................................................... 33 3.6 Risco oncológico................................................................................ 34 3.7 Seguimento Radiologico .................................................................... 40 JUSTIFICATIVA .............................................................................................. 42 HIPÓTESES ................................................................................................... 43 OBJETIVOS .................................................................................................... 44 REFERÊNCIAS .............................................................................................. 45 ARTIGO EM INGLÊS...................................................................................... 53

Inframammary Fold Reconstruction

Creating a well-defined inframammary fold is very important for the success of breast reconstruction, regardless of the surgical technique used. Inframammary fold reconstruction is mainly performed in the case of late postmastectomy breast reconstruction. Several surgical techniques have been described for both creation and correction or better definition of the inframammary fold. These techniques may be used during primary breast reconstruction or later, when the tissue expander is replaced with flaps, implants, or both. We describe four techniques for reconstructing the inframammary fold—external approach, internal approach, anchor approach, and allograft approach—as secondary procedures after breast reconstruction.