José Antônio Crespo Cavalheiro

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.

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.

Lipofiling Adds Diagnostic Difficulties in Imaging Evaluation After Radiotherapy? Case-Control Study

INTRODUCTION: Immediate autologous fat grafting has yet to be assessed as an alternative for breast reconstruction following conserving surgery. The present study sought to compare mammographically, patients undergoing conserving surgery with patients undergoing autologous adipose tissue immediately to conserving surgery, after completion of radiation. MATERIALS AND METHODS: This is a case-control study. The convenience sampling occurred between June 2010 and Dec 2011, where 28 patients coming from the outpatient Breast Care Unit with the indication of breast conserving surgery underwent breast reconstruction with fat grafting. Controls were consecutive cases, 60 patients, of conservative surgery that occurred in the same period. After the surgical procedure, all were subjected to radiotherapy. All patients underwent clinical and radiological evaluation at 6 months. RESULTS: The results of early evaluation in six months, showed no difference between the mammographic findings in patients undergoing conservative surgery and conventional surgical repair with fat grafting. CONCLUSION: Immediate autologous fat grafting is a promising technique for reconstruction after breast-conserving surgery. The evaluation performed by an observer who did not contain the information of the fat grafting technique has been shown that increased degree of harder it not for the diagnosis of changes in mammographic post surgery patients submitted to this technique. KEYWORDS: Autologous fat grafting; breast-conserving surgery; breast reconstruction; lipofilling; breast cancer.