OSNA Total Tumor Load for the Prediction of Axillary Involvement in Breast Cancer Patients: Should We use Different Thresholds According to the Intrinsic Molecular Subtype? MOTTO Study.

Aims: To assess the impact of the molecular subtype (MS) on the total number of CK19 mRNA copies in all positive SLN (TTL) threshold, to predict non-SLN affectation, and to compare 5 years progression-free survival (PFS) according to the risk of recurrence (ROR) group by PAM50. Methods: Cohort with infiltrating breast cancer with intra-operative metastatic SLN detected by one-step nucleic acid amplification (OSNA) assay who underwent subsequent ALND. Logistic regression was used to assess a possible interaction between TTL and MS(Triple Negative, Her-2-Enriched, Luminal A, or Luminal B), or hormone receptors (HR: positive or negative) by immunohistochemistry (IMH). Cox regression was used to compare PFS and OS in the 3 ROR groups (high, medium, or low). Results: TTL was predictive of non-SLN affectation in both univariate (OR [95% CI]: 1.72 [1.43, 2.05], P < .001) and multivariate (1.55 [95% CI: 1.04, 2.32], P = .030) models, but MS-IMH or HR-IMH, and their interactions with TTL were not (best multivariate model: HR + main effect OR 1.16 [95% CI: 0.18, 7.64], P = .874; interaction OR: 1.04 [0.7, 1.55], P = .835; univariate model: HR + main effect OR: 1.44 [95% CI: 0.85, 2.44], P = .180). PFS was lower in the high-risk ROR group (81.1%) than in the low-risk group (93.9%) (HR: 3.68 [95 CI: 1.70, 7.94], P < .001). Conclusions: our results do not provide evidence to support the utilization of subtype-specific thresholds for TTL values to make therapeutic decisions on the axilla. The ROR group was predictive of 5 years-PFS.

Human Papillomavirus in the region of La Ribera-Valencia: Present and future.

OBJECTIVE: Human Papillomavirus (HPV) is the main cause of cervical cancer. The etiology and effects derived from this infection are set by molecular techniques and cytological diagnosis, respectively. In the present study, data obtained by an opportunist screening of cervical cancer in La Ribera region are revised and related statistically. METHODS: Data considering different variables such as age, degree of lesion, HPV type detected and number of virus in coinfection were collected from 1,372 HPV positive cytology samples. HPV detection was carried out by means of three molecular techniques and the degree of lesion was analyzed by cytological diagnosis (Bethesda). In order to determine the relationship between different selected variables, several statistical analyses were performed. RESULTS: Only degree of lesion variable showed a direct relationship with the rest of variables, increasing with aging process, viral oncogenicity, presence of at least one high-risk virus and with the fact of being mono-infected. The probability of presenting a higher-level degree of lesion multiplied by 28.4 when high-risk HPV was detected in mono-infection. CONCLUSIONS: HPV molecular detection is the most suitable technique to perform a cervix cancer primary screening for the management of women with negative cytological diagnose. The number of detected types is statistically related to the degree of lesion. The establishment of a properly regulated screening to identify HPV infection, and therefore, of cervical cancer risk, is essential.

Elaboration of a nomogram to predict non sentinel node status in breast cancer patients with positive sentinel node, intra-operatively assessed with one step nucleic acid amplification method.

BACKGROUNDS: Tumor-positive sentinel node(SLN) biopsy results in a risk of nonsentinel node metastases in case of micro and macro metastases ranging from 20 to 50 %, respectively. Therefore, most patients underwent unnecessary axillary lymph node dissections. Thus, the development of a mathematical model for predicting patient-specific risk of non sentinel node(NSLN) metastases is strongly warranted. METHODS: The following parameters were recorded: CLINICAL: hospital, age, medical record number Bio-pathological: tumor (T) size, grading (G), multifocality, histological type, LVI, ER-PR status, HER-2, ki67, molecular classification (luminal A, luminal B, HER2 like, triple negative) Sentinel and nonsentinel lymph node related: number of removed SLNs, number of positive and negative SLNs, copy number of positive sentinel nodes, ratio: number of positive SLNs to number of removed SLNs, number of removed and number of positive nodes after ALND. A total of 2460 patients have been included in the database. All the patients have been provided by the authors of this paper. RESULTS: Multivariate logistic regression analysis demonstrated that only the number of a CK19 mRNA copies (p < 0.0001), T size (p < 0.0001) and LVI (p < 0.0001) were associated with NSN metastases. The discrimination of the model, quantified with the area under the receiver operating characteristics curve, was 0.71 (95 %, C.I. 0.69-0.73), thus confirming a good level of reliability. CONCLUSIONS: The nomogram may be employed by the surgeon as a decision making tool on whether to perform an intraoperative axillary lymph node dissection on breast cancer patients with SLN positive. The large population employed and the standardized method of measuring the value of CK19 mRNA copies are appropiate prerequisites for a reliable nomogram.

Molecular analysis of sentinel lymph nodes and search for molecular signatures of the metastatic potential of breast cancer.

Molecular assays are a new and invaluable tool in the assessment of axillary lymph node status and metastatic potential of breast cancer. Many protocols for assessing the sentinel lymph node (SLN) status have been developed based on cytology and/or histology, showing that the rate of detection of metastasis increases with the number of histologic sections examined and with use of immunohistochemical staining in addition to conventional Hematoxylin & Eosin staining. However, full standardization of protocols for this procedure has not been achieved. Further attempts to increase sensitivity and specificity of sentinel node analysis include molecular biology-based techniques such as the real-time polymerase chain reaction (RT-PCR) and, more recently, one step nucleic acid amplification (OSNA). The latter technique, that has sensitivity close to 100% and extremely high specificity along with good reproducibility, allows analysis of the SLN in full with an intraoperative procedure in approximately 30 minutes. This highly standardized method permits to compare results between groups and predicts the probability of involvement of the remaining axillary lymph nodes based on the total tumor load of the SLN(s). Results of multicenter clinical trials suggest that OSNA allows a better personalization of patients' care based on the results of SLN analysis, because it offers criteria to select patient with metastatic SLN who will not receive additional benefit from axillary clearance. Due to the current controversy on the best treatment of the axilla after a positive SLN, the SLN copy number of CK19 mRNA can have a high impact on therapeutic decisions in this group of patients. Breast cancer is a highly heterogeneous group of diseases, characterized by remarkable differences in the histopathological features, response to treatment and clinical outcome. Most of the clinical and translational research efforts during the last decades aimed at identifying markers that would allow to predict the metastatic potential of early breast cancer, and hence to assess accurately its prognosis and to inform the choice of adjuvant systemic treatments. It is now clear that neoplastic transformation, tumor progression and response to treatment are driven and accompanied by the deregulated expression of hundred or thousand genes, whose status cannot be assessed by the currently established histopathological and immunohistochemical approach. The new molecular assays have elicited a great deal of expectations, and for the most part they have been enthusiastically welcomed as potentially offering new chances for a better and more personalized care of the patients. Many, however, are still reluctant to consider these assays ready for use in the clinical practice, and keep waiting for a confirmatory evidence of their utility when the results of ongoing clinical trials will be mature.

Assessing the reproducibility of the microscopic diagnosis of sessile serrated adenoma of the colon.

INTRODUCTION: sessile serrated adenoma (SSA) is a recently described lesion that may be related to the development of up to 15% of colorectal cancers (CRCs). OBJECTIVE: to determine the accuracy of morphological criteria for the diagnosis of SSA by assessing concordance between pathologists. MATERIAL AND METHODS: concordance between two pathologists in the diagnosis of serrated lesions of the colon was studied for 195 lesions (187 hyperplastic polyps and 7 serrated adenomas). Size, location, morphology, and sampling method were collected of each lesion. Both pathologists were unaware of the previous diagnosis, macroscopic characteristics, and location of lesions. Possible diagnoses were: SSA, traditional serrated adenoma (TSA), hyperplastic polyp (HP), serrated polyp, tubular adenoma, or mixed lesions. Diagnostic doubts had to be described. Concordance between both observers was assessed using the kappa index (ê). The influence of collected variables on concordance degree was also evaluated. RESULTS: overall agreement on the histological diagnosis was poor (ê = 0.14), and so was agreement on the diagnosis of SSA (ê = 0.23). Concordance in the diagnosis of SSA improved with size > 5 mm (ê = 0.64) and proximal location (ê = 0.43). CONCLUSION: in a real clinical setting, the existing morphological criteria for SSA identification may be difficult to use.

Doxorubicin and cyclophosphamide followed by weekly docetaxel as neoadjuvant treatment of early breast cancer: analysis of biological markers in a GEICAM phase II study.

INTRODUCTION: To evaluate the sequential administration of doxorubicin (A) and cyclophosphamide (C) followed by weekly docetaxel in women with stage II to IIIA breast cancer. PATIENTS AND METHODS: Patients received 60 mg/m(2) of A and 600 mg/m(2) of C every three weeks for four cycles followed by 12 infusions of weekly docetaxel at a dose of 36 mg/m(2) and with a 2-week resting period. RESULTS: Sixty-three women were included. On an intention-to- treat basis, clinical response rate was 90% (95% CI: 83-98), with 46% complete responses. Breast-conserving surgery could be performed in 43 patients (68%). Complete pathological responses in the breast were confirmed in 17% of patients. No correlations between levels of expression of topoisomerase II alpha, survivin or p27 and the pathological response were detected. The study treatment was generally well tolerated. CONCLUSION: Neoadjuvant AC followed by weekly docetaxel is a feasible regimen for patients with early-stage breast cancer.

[Study of the sentinel node in breast cancer using lymphoscintigraphy and a fast method for cytokeratin]. / Estudio del ganglio centinela en el cáncer de mama mediante linfogammagrafía y citoqueratina por congelación.

INTRODUCTION: Histopathological examination of the axillary sentinel node (SN) is becoming a routine procedure in the surgical phase of infiltrating ductal carcinoma of the breast (IDC). The SN exam may yield false negative cases mainly due to identification failure of the SN but some of the false negative cases may be the result of the pathological examination procedure applied. MATERIAL AND METHODS: Sixty two (62) cases of clinically staged N0 IDC of the breast by TNM nomenclature were assigned to breast surgery along with conventional axillary node dissection. The identification technique included lymphoscintigraphy and intraoperative gamma-detecting probe after peritumoral injection of 99mTc-labeled colloids.The histological study of SN was performed with paired 4 microm slices and staining with hematoxylin-eosin and with a fast method of cytokeratins for freezing. RESULTS: In only two of the 62 patients, it was not possible to identify the SN. Eighteen of the remaining 60 had SN involvement by metastasis, having no metastases in the other nodes of the axillary dissection in 6 of them. Ten of those were micrometastasis (size of metastasis= or <0.2 cm). In two out of these last 10 cases, diagnosis of the micrometastasis was only possible using slices stained with CK. There were no false negative results. CONCLUSIONS: The lymphoscintigraphy, after peritumoral injection of small volumes and low dose of the tracer, makes it possible to obtain excellent results in the intraoperative detection of the SN in breast cancer. The study of this SN with a fast method for CK decreases the number of false negative results of the technique.

Characterization of GIST/GIPACT tumors by inmunohistochemistry and exon 11 analysis of c-kit by PCR.

Gastrointestinal stromal tumors (GIST/GIPACT) are the most common mesenchymal tumors of the human gastrointestinal (GI) tract and are characterized by the constant immunohistochemical expression of CD117. In recent years, sporadic and germ line mutations in the c-kit gene have been described in GIST/GIPACT tumors, resulting in a constitutive activation of the gene. The most prevalent mutation is located in exon 11 of the c-kit gene, involved in the transcription of the juxta-membrane domain of the c-kit protein. There are conflicting reports with respect to the association between exon 11 mutations and the biological behavior of GIST/GIPACT tumors. This work studies eight patients with tumors diagnosed as GIST/GIPACT, both morphologically and immunohistochemically for CD117, CD34, a-smooth muscle actin, desmin and S-100 protein primary antibodies. The DNA of the eight cases was also studied by PCR for mutation of exon 11 of the c-kit gene. All cases were CD117 positive, but only two showed mutation of exon 11. These last two cases did not show morphological characteristics of malignancy. The most aggressive case, with early death of the patient, did not show the mutation. In conclusion, there was no correlation between the mutation of exon 11 of the c-kit gene and the malignant behavior of GIST/GIPACT tumors in our series.