Cell morphology impact on the set-up of electroporation protocols for in-suspension and adhered breast cancer cells.

In order to establish cancer-type-specific electroporation protocols for breast cancer, electroporation was performed in vitro in two modalities: in-suspension and adhered cells. Electroporation of cell suspensions was carried out through commercial electroporation cuvettes whereas a novel electrode for electroporation of adhered cells was designed and manufactured aimed to preserve cell structure, to provide a closer model to an in vivo scenario, and as a means to visualize the mechanical effects of electroporation on the cell membrane by using scanning electron microscopy. Electroporation protocols and electric field thresholds were predicted in silico and experimentally tuned through propidium iodide uptake and cell viability. Three breast-cancer cell lines (BT-20, MCF-7 and HCC1419) and a non-cancerous cell line (BEAS-2B) were used. Cancerous cells responded differently to electroporation depending on the electric parameters, cell histology, the cell culture modality, and the cell morphology (membrane thickness mainly), which was evaluated trough confocal and transmission electron microscopy. Particularly, it was found that electrochemotherapy may represent a promising alternative as an adjuvant treatment of metastatic breast tumours, and as a neoadjuvant therapy for Her2/neu tumours. Oppositely, triple negative breast tumours may show a high sensitivity to electroporation and therefore, they could be efficiently treated with irreversible electroporation. On the other hand, noncancerous cells demanded the highest voltage in both cell culture modalities in order to be electroporated. Hence, these cells in suspension may provide a reliable, easy-to-perform, low-cost model for the development of electroporation protocols for eradication of healthy tissue around a tumour in a safety margin.

[Metaplastic carcinoma of the breast and the impact of the p63 and cytokeratin 5/6: experience of 40 patients]. / Carcinoma metaplásico de mama y repercusión de la expresión de p63 y citoqueratina 5/6: experiencia de 40 pacientes.

BACKGROUND: Metaplasic carcinoma of the breast was initially described by Huvos in 1974. It is a rare and aggressive entity characterized by the presence of mesenchymal and epithelial components. OBJECTIVE: To know the incidence and biologic behaviour of the metaplasic carcinoma of the breast at the Instituto de Enfermedades de la Mama, FUCAM, AC. METHODS: Data on women diagnosed with metaplasic carcinoma of the breast between January 2005 and December 2014 was collected by retrospectively reviewing in FUCAM. Clinical, pathological and immunohistochemical characteristics were assessed. The five-year disease-free survival (DFS) and overall survival (OS) were evaluated. RESULTS: a total of 4198 patients have been diagnosed with breast cancer in our institution, 40 (0.95%) of them with metaplasic carcinoma. The median age of the patients was 46 years (27-73). 60% of the patients were diagnosed with an advanced clinical stage (III) and the triple-negative subtype was the most frequently found. A mean follow-up of 24 months showed rates of overall survival and disease-free survival of 80% and 69.9%, respectively. The presence of both, cytokeratins 5/6 and p63, seems to have a negative impact in local recurrence. CONCLUSION: this study demonstrates that metaplasic carcinoma is a rare and aggressive disease. Expression of both tumor cytokeratins was associated with a worse outcome.

[Breast Cancer Survival: Clinical andPathological Prognostic Factors Analysis]. / Supervivencia de pacientes con cáncer de mama. Análisis por factores pronóstico, clínicos y patológicos.

Background: Breast cancer is the leading cause of cancer death in women in Mexico, is a heterogeneous disease, and knowledge of prognostic factors are critical in making treatment decisions. Objetive: determine the overall survival (OS) and disease-free survival (DFS) at 5 years, analyzed by risk groups. Material and methods: Patients diagnosed with breast treated at the Institute of Breast Diseases FUCAM from July 2005 to December 2014 were included. Simple frequencies were used for analysis of the general characteristics, and 5- year OS and DFS were analyzed using Kaplan-Meier curves. A subset analysis of the clinical stage and comparing survival in those patients diagnosed by mammography screening program was performed. Results: 4,902 patients with breast cancer were included, general clinical and pathological features are described and 3,762 patients were included for analysis of 5-year OS and DFS. The average age at diagnosis was 53.7 years; 13.3% were <40 years, which deleteriously reflects on the supervivencia global 76 vs 84% in >40. At diagnosis predominated locally advanced stages (45%), OS and DFS at 5 years was 96.8 ± 0.6% and 93.4%±0.9 respectively for early stages, 74.6 ± 1.7% and 68.7 ± 2% for locally advanced and 35.9 ± 5.1% and 37.4 ± 10.3% for metastatic tumors. Women diagnosed in the screening program had significantly better OS and DFS compared with symptomatic patients (95 and 93% vs 79 and 77%). For biological subtypes, OS and DFS was 89 and 84% for luminal, 81 and 81% for luminal Her +, 74 and 78% for pure Her 2, and 69 and 73% for triple negative. Conclusion: Knowledge of the prognostic factors that affect survival of patients with breast cancer is essential for categorizing risk groups and to individualize treatment in order to improve life expectancy.

Tumor size as predictor of microinvasion, invasion, and axillary metastasis in ductal carcinoma in situ.

Ductal carcinoma in situ (DCIS) represents a small number of cases in countries with inadequate breast cancer screening programs, and in the majority of cases is diagnosed as a palpable lump. It has been proposed that DCIS with palpable lump > or = 2.5 cm can be associated with microinvasion or invasive carcinoma and risk of axillary metastasis. The purpose of the present study is to evaluate incidence of microinvasion, invasion, and the role of lymphatic mapping and sentinel lymph node biopsy in DCIS > or = 2.5 cm. We conducted a retrospective analysis of patients with histologically proven incisional, excisional, or core biopsy of DCIS lump > or = 2.5 cm at a tertiary-care hospital. All patients underwent lymphatic mapping with sentinel lymph node biopsy.A total of 24 patients were included with average tumor size of 4 cm (range, 2.5-6 cm); 29% had microinvasive and 12.5% had invasive disease, three patients (12.5%) had positive sentinel lymph node, all had micrometastasis, and no metastasis were found in non-sentinel lymph nodes. Incidence of microinvasion and invasion were directly related with tumor size (10% for DCIS tumor size of 2.5-3.5 cm, 57% for 3.6-4.5 cm, and 71% for tumors between 4.5 and 6 cm). In addition, axillary metastasis incidence had a direct relationship with tumor size. (0% in 2.5-3.5-cm tumor size, 14% for 3.6-4.5 cm, and 28% in DCIS between 4.6 and 6.0 cm). The present study shows high incidence of microinvasion and invasion in DCIS diagnosed in tumors > or = 2.5 cm and supports the importance of axillary evaluation in patients with tumors >3.5 cm by means of lymphatic mapping and sentinel lymph node biopsy.

A case report of mucoepidermoid carcinoma of the parotid gland developing after radioiodine therapy for thyroid carcinoma.

This is a report on a 19-year-old female who developed a low grade T2 N0 M0 mucoepidermoid carcinoma of the right parotid gland 3 years and 5 months after the post-operative treatment of 100 mCi of radioactive iodine (131I) for a papillary thyroid carcinoma. The parotid tumour appeared during the patient's pregnancy. There are few reports of salivary gland cancer developing after radioiodine therapy for thyroid carcinoma and it is hoped that this report may stimulate others to investigate this association further in order to clarify the risk of secondary malignancies after radioiodine therapy.

Prognostic significance of cutaneous depigmentation in Mexican patients with malignant melanoma.

BACKGROUND: The clinical significance of cutaneous depigmentary phenomena in patients with malignant melanoma is not clear. There are two varieties: 1) vitiligo (VIT), and 2) leukoderma acquisitum centrifugum (LAC). METHODS: In order to evaluate the outcome of our patients with malignant melanoma and skin depigmentation (VIT or LAC), the patients in this study with this association were retrospectively reviewed and compared with the total melanoma patients at the Oncology Hospital and the XXI Century National Medical Center of the Mexican Social Security Institute in Mexico City. RESULTS: Nine cases were found from 1985-1995. There were eight women and one man, their mean age was 63 years. Six melanomas were located in the foot, one in the leg, one in the anus and one in the neck. All were Clark's levels III, IV or V, and their mean tumor thickness was 5.7 mm. Four out of nine patients had regional lymph node metastasis. Six melanomas were associated with VIT and three with LAC. Three patients developed the depigmentation after chemo- or chemoimmunotherapy. All nine patients are alive (100%) with a mean follow-up of 55 months (9-141), eight out of nine have no evidence of tumor. CONCLUSIONS: From these data it may be concluded that the study patients with malignant melanoma and VIT or LAC have a higher-than-expected survival according to their prognostic factors. Therefore, the presence of the depigmentation phenomena must be looked for intentionally.

Electrochemotherapy in primary and metastatic skin tumors: phase II trial using intralesional bleomycin.

BACKGROUND: Surgery and/or radiotherapy are the elective therapies for most primary skin cancers. Nevertheless, some patients develop recurrences, and chemotherapy has resulted in poor complete responses. Permeabilization of the cell membrane by electric pulses allows bleomycin to enter into the cell, increasing possibility of cytotoxicity. METHODS: From November 1998 through November 1999, 15 patients with 38 skin lesions participated in a phase II prospective clinical trial, using intralesional bleomycin plus electric pulses delivered 10 min after bleomycin injection, which lasted 100 microsec each at field strength of 1,300 V/cm and a frequency of 1 Hz. There were basal cell carcinomas (BCC) (nine lesions), in-transit metastasis of melanoma (MM) (two patients/13 nodules), squamous cell carcinomas (SCC) of the upper aerodigestive tract metastatic to the skin (two patients/two nodules), and skin metastases from breast cancer (two patients/14 nodules). Mean follow-up was 8.6 months. RESULTS: Overall objective responses were 98%, with complete responses achieved in 49%, partial responses in 49%, and no responses in 2%. No complications were documented related to the treatment and tolerance was adequate. CONCLUSIONS: Electrochemotherapy (ECT) is a new cancer modality of treatment that is effective in a variety of skin cancers. This treatment represents an excellent alternative to standard surgery or radiotherapy, with an outpatient-based treatment applied in one to three sessions. The major impact was obtained in BBC, but ECT is a useful palliative therapy in melanoma, breast cancer, or SCC. More experience and longer follow-up are required to determine long-term results.

Quality control in planning and technique of radiotherapy with cobalt-60 for T1 glottic cancer increase local control and organ preservation.

BACKGROUND: In order to evaluate whether individualized technique and dosimetry of radiotherapy increase local control, organ preservation, and survival of patients with T1 glottic cancer, we reviewed 76 cases treated from 1979 to 1993. METHODS: Group A included 32 patients treated from 1979 to 1989 with different techniques, based on clinical aspects. Group B included 44 patients treated from 1990 to 1993 with individualized technique according to tumor extension and patient's anatomy. RESULTS: Five-year local control with radiotherapy alone was achieved in 53% of group A versus 91% of group B (P > 0.005). Survival was similar in both groups with rescue surgery (90% versus 96%). Five-year survival with larynx preservation was 65% in group A versus 88% in group B (P = 0.02). Most recurrences (78%) appeared within 24 months of follow-up. CONCLUSION: Adequate staging, individualized technique, computing planning using simulation and use of immobilization devices during cobalt-60 radiotherapy significantly increase local control and organ preservation in T1 glottic cancer.

A simplified histoscore for the estrogen receptor assay in breast cancer.

Different histoscores combining the number of positive cells and the intensity of staining have been used to evaluate the estrogen receptor immunocytochemical assay (ER-ICA). Our aim was to investigate if the simple estimation of the amount of positive cells could be sufficient for the semiquantitative analysis of ER-ICA. Tissue from 51 women with ductal breast carcinoma was used. Half of each sample was processed with the quantitative assay (ER-EIA) and the other half with ER-ICA. Microscopical analysis was performed by two independent observers and classified on a simple scale from 0 to 4+. With EIA 31 cases (60.78%) were positive and 20 (39.21%) negative. With ER-ICA 29 (56.86%) had immunostaining, whereas 22 (43.13%) did not. 95.83% of the ER-ICA positive cases and 77.7% of ER-ICA negative had a good correlation with EIA values. Statistical analysis showed a high degree of correlation (r = 0.88 p 0.001). Hence, simple semiquantitative estimation in ER-ICA is sufficient to provide useful information for clinical use about ER content in tissue sections.

Multifactorial analysis of survival and recurrences in differentiated thyroid cancer. Comparative evaluation of usefulness of AGES, MACIS, and risk group scores in Mexican population.

Many risk factors have been identified in differentiated thyroid cancer, with them, some prognostic scores have been designed to asign the risk of recurrence and survival. In Mexican population, this type of study is scarce. This is a retrospective review of 180 patients with differentiated thyroid cancer completely treated at the Hospital de Oncologia, IMSS, in Mexico City from 1980 to 1990. All prognostic factors were analyzed and a score obtained either by the method of AGES, MACIS, or SKMH. Correlation of recurrences and survival was carried out according to score or risk assignment. There was a predominance of females (4.8:1), 48% had metastatic cervical nodes, median tumor size was 4 cm, 16% had multiple macroscopic thyroid tumors, in 12% resection was incomplete, 96% were papillary, and 4% follicular cancers. According to AGES, 46% were high risk patients, 49.4% with MACIS and 45.5% with SKMH, respectively. Median follow-up was 8.3 years. There were 67 (37%) recurrences. Ten-year overall survival was 89.4% and disease-free survival was 79.2%. There was no statistical significant difference of survival of AGES until the score reached 6 or more or the MACIS score reached 8 or more. Cox multivariate model showed that above the age of 45, tumor size of 5 cm or more, follicular histology, multiple macroscopic thyroid tumors, and extracapsular node invasion affected ten-year survival. In conclusions our patients are diagnosed at more advanced stages than patients in the U.S. or European countries. Nearly one half of our patients belonged to the high-risk group. This study confirms that patients over the age of 45, tumor size > 5 cm, and follicular histology are adverse prognostic factors and report that extracapsular node invasion and multiple macroscopic thyroid tumors are also adverse prognostic factors. In Mexican population, with 50% of high-risk patients, AGES and MACIS scores reached statistical differences with higher qualifications than observed in the U.S.

Evaluation of a simple semiquantitative assay for estrogen receptors in breast cancer.

A number of studies have been published to evaluate the inmunocytochemical assay for ER using monoclonal antibodies. Histoscores so far used consider two variables: the number of cells and the intensity of the reaction. There are however indications that only the proportion of stained cells are important for assessment and show a direct correlation with quantitative data. We studied 77 breast invasive adenocarcinomas stained with the Abbott ERICA kit and used a simple scale of 0 to 4 for the estimation of ER. Tissues were snap frozen in liquid N and immunostained with the specific antibody and peroxidase. Immunostaining was estimated in a simple observational scale from 0-4+ where 0 = no staining or few scattered positive cells; 1+ up to 25%; 2+ up to 50%; 3+ up to 75% and 4+ more than 75% of stained malignant cells. Counts were performed in at least 100 malignant cells in various microscopic fields. Staining was always nuclear and a considerable heterogeneity in the number of cells and the intensity of the reaction was observed. Grading specimens from 0-4+ was found simple and reproducible. In 45% there was no immunostaining and 54% were positive for ER. In patients greater than 50 years of age 67% were positive; in patients less than 50 years of age only 33% had ER. This procedure has many advantages for clinical use: it is simple, it does not require sophisticated equipment, it is reproducible and can be performed in small tissue fragments, such as needle aspiration material, as well as in cytological smears.

[Breast cancer in Mexico. Is it a young women disease?]. / Cáncer de mama en México. Enfermedad de mujeres jóvenes?

Breast cancer in México has the second place in frequency in general population (10.6%) as well as in females (16.4%). There has been found an increase in mortality from 3.6, in 1985, to 6 x 100,000 inhabitants, in 1994. Breast cancer diagnosis is made in advanced stages (III, IV, N.C.) in two out of three of our patients and a great proportion of them are younger than 50 years old. In order to know if age at diagnosis of breast cancer women in Mexico is similar of that of women of other countries, and in consequence, if breast screening has to be done as they have determined, we analyzed the age at diagnosis of 29,075 mexican women with breast cancer from 1993 to 1996. We found that median age of our patients was 51 years, one decade younger than that of European or white American women (60-64 years), and 45.5% of Mexican women are less than 50 years old at diagnosis of breast cancer, therefore, breast cancer screening indications adopted in Mexico, do not cover, not even theoretically, almost 50% of our women. It is mandatory to determine indications of breast screening according to our reality.

[Reintervention to complete the surgical treatment of thyroid cancer. Indications and histopathological findings]. / Reintervención para completar el tratamiento quirúrgico en cáncer de tiroides. Indicaciones y hallazgos histopatológicos.

In Mexico, 39% of 158 patients operated on for thyroid cancer require reoperative thyroid surgery. We retrospectively reviewed the indications and histopathological findings of 60 patients reoperated on because of: a) suspected persistent or recurrent disease; b) high risk patients treated by lobectomy; c) different histology; d) complete lack of information, e) and distant metastasis. In 53 cases (88%), the initial surgery was nodulectomy or lobectomy, and in seven (11%) was subtotal or near-total thyroidectomy. Among the 60 reoperations, 50 were completion total thyroidectomy and 10 were near-total thyroidectomy. In 27 cases (45%) a neck dissection was additionally done. Histologic examination revealed thyroid carcinoma in 32 cases (53%) and neck node metastasis in 28 cases (47%). Complications included six cases (9%) of permanent palsy of the recurrent laryngeal nerve after the initial surgery outside of our hospital and two cases (1.75%) of reoperated cases. In four reoperated patients (6.6%), permanent hypoparathyroidism was developed. It is mandatory to complete thyroidectomy and neck dissection in a high proportion of patients initially treated in general hospitals due to an inadequate criteria in the selection of the extension of thyroidectomy and treatment of neck node metastases. Histologic findings of these patients support our indications to complete the surgical treatment.

Cancer of the larynx in Mexico: review of 357 cases.

Descriptions of the patient population suffering from carcinoma of the larynx are not common in Mexico. This article deals with the clinical features, treatment, and results of 357 cases of cancer of the larynx treated at the Hospital de Oncologia, Centro Medico Nacional, IMSS, during a 10-year period. In this series, 68% of the patients were in advanced stage (T3-T4) at the time of diagnosis; nevertheless, a combination of surgery and radiotherapy achieved an actuarial 5-year survival for the entire group of 77.5%. Metastatic cervical nodes and tracheotomy previous to the treatment were factors that influenced a poor prognosis (p = 0.01). The survival of cases in the early stages (T1-T2) of 95.2% is comparable to that reported by most authors. In 87.5% of the cases that develop recurrence, it appears during the first 24 months of control.

Carotid body tumors in inhabitants of altitudes higher than 2000 meters above sea level.

BACKGROUND: Carotid body tumors (CBTs) are rare. Diagnosis is usually delayed until the tumors reach a critical volume and a mass appears in the neck, which is often asymptomatic. METHODS: We reviewed retrospectively the cases with CBT diagnosed from 1965 to 1995 in the Hospital de Oncologia in Mexico City with inhabitants at an altitude higher than 2200 m above sea level to see whether these tumors have the same characteristics as those of inhabitants of countries of lower altitudes. We reviewed the clinical features, diagnostic procedures, therapy, results, and complications. RESULTS: There were 120 CBT cases, which represent 79% of the parapharyngeal space tumors diagnosed at our hospital; 116 (96%) were benign and 4 (3.3%) were malignant. Women predominated (89%), and the female-male ratio was 8.3:1. Five patients had multiple paragangliomas, and one had a family history of CBT. Eighty patients (66%) underwent surgery. This was done by cervical approach in 78 cases (97%), and 2 (2.5%) required additional mandibulotomy. There were three deaths due to brain ischemia after carotid ligature. Forty-one patients were followed without treatment, due to advanced age, concomitant diseases, or great volume of the tumor. Median follow-up of these patients was 47 months, during which time no patient reported additional symptoms, accelerated enlargement of the tumor, or metastasis. With a median follow-up of 54 months, only one patient developed local recurrence and three patients developed distant metastasis. CONCLUSIONS: We conclude that cases of CBT in our high-altitude population differ significantly from those cases in inhabitants of cities in the U.S. or Europe of less than 1500 m above sea level. Those of high altitudes have an evident female predominance (8.3:1), low rate of bilaterality (5%), and a family history of 1% versus a discrete female predominance (2:1), bilaterality from 10% to 20%, and family history from 7% to 25% in low altitudes. When adequate criteria are used to determine surgical resectability, a complete resection is achieved in 85% of cases, with low or null mortality and high local control.

Oral feeding after total laryngectomy for endolaryngeal cancer.

There is still no agreement about the beginning of oral feedings after total laryngectomy. Some authors begin routine feedings on the 3rd postoperative day, while others delay oral feedings until 12-14 days after surgery. The present study was devised as a prospective randomized clinical trial concerning beginning oral feedings on the 7th or 14th postoperative day in 35 patients following total laryngectomy as treatment for endolaryngeal cancer. There were no differences in risk factors present in either group. Only two pharyngocutaneous fistulae occurred in the overall series (5.7%), with both appearing in the 7-day group and developing on the 18th and 20th postoperative days respectively. There were no statistically significant differences in fistula formation between the two groups (P = 0.49). We conclude that it is not necessary to delay oral feedings more than 7 days in patients without suture line tension.