Breast cancer cutaneous metastases are associated to uMUC1 and sialyl Lewis x and to highly malignant primary tumors.

Breast cancer spreading to different organs have been related to different molecules and mechanisms, but cutaneous metastasis remains unexplored. Increasing evidence showed that MUC1 and some of its carbohydrate associated antigens may be implicated in breast cancer metastasis. In this study we analyzed these tumor markers in order to identify breast cancer cutaneous metastatic profiles. A cohort of 26 primary tumors from breast cancer patients with cutaneous metastases were included; also, cutaneous and lymphatic node metastatic samples and primary tumors from breast cancer patients without metastases were analysed. Immunohistochemical (IHC) studies demonstrated that both underglycosylated MUC1 (uMUC1) and sialyl Lewis x (sLex) to be positively associated with cutaneous metastatic primary tumors (p < 0.05). Notably, a high percentage of tumors with cutaneous metastases were characterized as triple negative and Her2+ tumors (37.5 % and 29 %, respectively). Some discordant results were found between primary tumors and their matched cutaneous metastases. To determine if MUC1 variants may be carriers of carbohydrate antigens, subcellular fractions from a cutaneous metastatic lesion were obtained, immunoprecipitated and analyzed by Western blot. We found that the isolated uMUC1 with a molecular weight of>200 kDa was also the site for binding of anti-sLex MAb; in coincidence, a high correlation of positive IHC expression of both markers was observed. Our findings confirm that breast cancer cutaneous metastases were associated to highly malignant primary tumors and sustain the hypothesis that u-MUC1 and sLe x may drive breast cancer cutaneous metastases.

Generatividad y Satisfacción Vital en un una muestra de abuelos/as de Santiago, Chile / Generativity and Life Satisfaction in a sample of grandparents from Santiago, Chile

Resumen Se presentan los resultados de una muestra de 385 abuelos1 chilenos estratificada por sexo, edad y nivel de ingreso del grupo familiar en la Escala de Generatividad de Loyola (McAdams & de St. Aubin, 1992) adaptada para abuelos en Chile (EGL-A) por Larrain, Zegers & Orellana (2016) y en el Índice de Satisfacción Vital de Neugarten, Havighurst & Tobin (1961,1996) en su versión adaptada a Chile por Zegers, Rojas-Barahona & Föster (2009). No se encontró evidencia de correlación positiva entre los puntajes totales de ambos instrumentos, pero sí al estratificar por escolaridad, la correlación entre generatividad y la dimensión congruencia del LSI-A resultó positiva para quienes poseían escolaridad media (r = 0.215, p= 0.021) o superior (r = 0.230, p= 0.0066).

Abstract The results of a sample of 383 chilean grandparents stratified by sex, age and income level to the Loyola Generativity Scale (LGS) (McAdams & de St. Aubin, 1992) adapted for grandparents in Chile (EGL-A) by Larrain, Zegers & Orellana (2016) and the Life Satisfaction Index Neugarten, Havighurst & Tobin (1961, 1996) in its adapted version to Chile by Zegers, Rojas-Barahona & Föster (2009) are presented. It was not found a positive correlation between the total scores of both instruments but it was positive when the sample was stratified by school level between generativity and the congruence dimensión of the LSI-A for all those with complete high school studies (r = 0.215, p= 0.021) or college studies (r = 0.230, p= 0.0066).

IDO is highly expressed in breast cancer and breast cancer-derived circulating microvesicles and associated to aggressive types of tumors by in silico analysis.

Indoleamine-2,3-dioxygenase (IDO) has been established as a normal mechanism of peripheral tolerance and immunosuppression. Besides, malignant tumors release microvesicles (MV) related with tumor dissemination. The aims of this study were to determine the expression of IDO in breast cancer and circulating microvesicles from breast cancer patients and to perform an in silico analysis to find genes co-expressed to IDO. One hundred and twenty-two tissue and serum breast samples (91 malignant, 21 benign, and 10 normal), and MCF7, MDA-MB-231, and T47D breast cancer cell lines were included. Standard immunohistochemistry (IHC), immunocytochemistry (ICC), Western blot (WB), and RT-PCR were employed. Microvesicle isolation from plasma samples was obtained by serial centrifugation and ultracentrifugation. By IHC, 60 % breast cancer, 43 % benign, and 20 % normal samples were positive. Significant differences were found among normal, benign, and malignant samples. Breast cancer stages I, II, and III expressed IDO in 42, 66, and 71 % of samples, respectively, while breast cancer cell lines also reacted; by WB, 9/25 microvesicles fractions showed bands at 42 kD. In silico analysis of IDO 1 gene expression in breast cancer showed its association with several genes related to immune response and apoptosis. Moreover, IDO and co-expressed genes were found predominately in basal and erbB2 subtypes. The cumulative data indicate a high expression of IDO in breast cancer which increased with higher stages. Furthermore, IDO was found in association with circulating breast cancer MV, while experimental and in silico gene expression revealed that IDO was mainly expressed in a triple-negative subgroup.

Rhomboid family gene expression profiling in breast normal tissue and tumor samples.

Rhomboid is an evolutionary conserved and functionally diversified group of proteins composed of proteolytically active and inactive members that are involved in the modulation of multiple biological processes such as epidermal growth factor receptor signaling pathway, endoplasmic reticulum-associated degradation, cell death, and proliferation. Recently, several human rhomboid genes have been associated with the development of chronic myeloid leukemia and pituitary, colorectal, ovarian, and breast cancers. In this study, we evaluated the mRNA and protein expression profiles of rhomboid genes in cancer cell lines and breast tissue/tumor samples. In silico analysis of publicly available gene expression datasets showed that different rhomboid genes are specifically expressed according to the breast cancer intrinsic subtypes. Quantitative reverse transcription-polymerase chain reaction (RT-PCR) analysis showed a significant RHBDD2 mRNA overexpression in advanced breast cancer compared with normal tissue samples (p = 0.012). In addition, we found that RHBDL2 and PARL mRNA expression was associated with a low/intermediate histologic tumor grade (p = 0.024 and p = 0.015, respectively). Immunohistochemistry analysis showed a significant increase of RHBDD2 protein expression in association with breast cancer samples negative for progesterone receptor (p = 0.015). Moreover, protein expression analysis corroborated the quantitative RT-PCR results, indicating that breast primary tumors belonging to patients with a more disseminated disease expressed significantly increased levels of RHBDD2 protein compared with less disseminated tumors (p = 0.01).

Hepatotoxicidad por verapamil: reporte de un caso / Liver injury due to treatment with verapamil: case report

Drug induced liver injury is an infrequent adverse effect of Verapamil. It has been reported in about ten cases since 1981. We report a case of a middle-aged woman who received Verapamil in a dose of 120mg/d for eight weeks, developing a mixed hepatocellular and cholestatic pattern of liver injury without any symptoms. There was no evidence of eosinophilia. Liver function tests returned quickly to normal levels after discontinuation. As in previously reported cases, we hypothesize that this is an idiosyncratic mechanism of liver injury.

La hepatotoxicidad es un efecto adverso infrecuente de verapamil, existiendo alrededor de 10 casos reportados desde 1981. Relatamos el caso de una mujer de edad media que presenta una alteración de las pruebas hepáticas con patrón mixto asociado al uso de verapamil 120 mg/d por 8 semanas sin síntomas asociados, la cual responde rápidamente a la suspensión de este fármaco. Al igual que en casos reportados previamente, planteamos como mecanismo de injuria una hepatotoxicidad idiosincrática por este medicamento.

Rhomboid domain containing 2 (RHBDD2): a novel cancer-related gene over-expressed in breast cancer.

In the course of breast cancer global gene expression studies, we identified an uncharacterized gene known as RHBDD2 (Rhomboid domain containing 2) to be markedly over-expressed in primary tumors from patients with recurrent disease. In this study, we identified RHBDD2 mRNA and protein expression significantly elevated in breast carcinomas compared with normal breast samples as analyzed by SAGE (n=46) and immunohistochemistry (n=213). Interestingly, specimens displaying RHBDD2 over-expression were predominantly advanced stage III breast carcinomas (p=0.001). Western-blot, RT-PCR and cDNA sequencing analyses allowed us to identify two RHBDD2 alternatively spliced mRNA isoforms expressed in breast cancer cell lines. We further investigated the occurrence and frequency of gene amplification and over-expression affecting RHBDD2 in 131 breast samples. RHBDD2 gene amplification was detected in 21% of 98 invasive breast carcinomas analyzed. However, no RHBDD2 amplification was detected in normal breast tissues (n=17) or breast benign lesions (n=16) (p=0.014). Interestingly, siRNA-mediated silencing of RHBDD2 expression results in a decrease of MCF7 breast cancer cells proliferation compared with the corresponding controls (p=0.001). In addition, analysis of publicly available gene expression data showed a strong association between high RHBDD2 expression and decreased overall survival (p=0.0023), relapse-free survival (p=0.0013), and metastasis-free interval (p=0.006) in patients with primary ER-negative breast carcinomas. In conclusion, our findings suggest that RHBDD2 over-expression behaves as an indicator of poor prognosis and may play a role facilitating breast cancer progression.

MUC1 cytoplasmic tail detection using CT33 polyclonal and CT2 monoclonal antibodies in breast and colorectal tissue.

UNLABELLED: The immunohistochemical detection (IHC) of MUC1-CT employing a polyclonal antibody (CT33) in relation to CT2 monoclonal antibody (MAb) was analyzed. Western blot (WB) was used to determine the molecular mass of CT. MATERIALS AND METHODS: We studied 163 breast and 89 colorectal cancer specimens, 10 breast and 14 colorectal benign conditions, and 12 breast and 20 colorectal normal samples. From each tumor sample, subcellular fractions were obtained and analyzed by SDS-PAGE and WB. A nonparametric statistical analysis was employed; data were standardized and a Kendall-Tau correlation was applied. RESULTS: By IHC, 146/163 (90%) and 151/163 (93%) of breast cancer were positive with CT33 and CT2, respectively; a statistically significant correlation was obtained (t=0.5199). Seven out of ten (70%) benign breast specimens were positive with CT33 while all samples stained with CT2; in normal breast sample tissues, all were positive with both Abs. In colorectal cancer samples, both antibodies stained 47/89 (53%) samples; CT2 reacted in 13/14 (93%) of benign samples while CT33 showed a positive reaction in 9/14 (64%) of benign specimens. In normal samples, CT2 showed staining in 17/20 (85%) of samples and CT33 was reactive in 12/20 (60%). By WB, in breast and colorectal cancer samples, similar results were obtained with both antibodies: a main band at about 30kDa which represents the smaller subunit. CONCLUSION: CT33 polyclonal antibody has demonstrated its efficacy to detect MUC1 in breast and colorectal cancer tissues with similar reactivity to CT2. It is worthwhile to affirm that CT33 is a good indicator of MUC1 expression.

[Study of the reineta protein modifications (Brama australis), put under freezing and storage to -18 degrees C and -30 degrees C]. / Estudio de las modificaciones en proteínas de reineta (Brama australis), sometidas a congelación y almacenamiento a -18-C y-30-C.

The objective of the present work was to study functional and thermal properties of reineta (Brama australis) frozen meat, analysed by water retention capacity (WRC), gel forming capacity (GFC), texture, emulsifying capacity and differential scanning calorimetry (DSC). For this study, reineta fillets were obtained and extracted by the same conditions, and cutted, packaged, frozen and stored at -18 degrees C and -30 degrees C for 7 months. The results obtained, showed that there were no signifficant differences in the responses to thermal treatment for all the specimens. For samples frozen at -18 degrees C and -30 degrees C, the protein contents were 23.5 + 0.0 and 25.4 + 1.0%, respectively. The WRC values were 0.45 + 0.1 and 1.59 +/- 0.0 g water/g protein, respectively. The gel forming capacity was only present in the fresh samples, whereas the frozen stored ones only form protein aggregates. The emulsifying capacity was between 960 and 1400 g oil / g protein, and the storage time increased this value. The miosin denaturation temperature (Td) and denaturation enthalpy (?H), obtained by DSC, fluctuated between 39.2 +/- 0.5 to 44.8 +/- 0.8 degrees C and 1.12 +/- 0.3 to 0.52 +/- 0.2 J/g, respectively. The actina values were between 71.0 +/- 0.6 to 75.3 +/- 0.5 degrees C and between 0.5 +/- 0.1 to 0.7 +/- 0.1 J/g. Cooperativity decreased as the storage time increased. This is showing a certain degree of protein displacement. The values found by thermal analyses showed a direct relationship with the functional properties, both decreasing with storage time.

Antigenic differences between metastatic cells in bone marrow and primary tumours and the anti-MUC1 humoral immune response induced in breast cancer patients.

The dissemination of a malignant neoplasia is a complex process, which requires a set of molecules that remains unknown. It has been suggested that mucins and their carbohydrate-associated antigens may be implicated in tumour spreading which may be also influenced by an anti-MUC1 immune response. In this pilot study, we report the pattern of carbohydrate and peptidic MUC1-associated epitopes on carcinoma cells isolated from bone marrow (BM), taking into account primary tumour histopathologic features. We also bring information about the anti-MUC1 humoral response in these patients. Seventeen patients with invasive breast carcinoma were included. A sample of the primary tumour, a serum sample and a BM aspirate were obtained from each patient. Clinical features studied were tumour size, number of metastatic nodes, histological type and disease stage. Standard immunohistochemistry was performed with antigenic retrieval using different monoclonal antibodies (MAbs): anti carbohydrate antigens: Lewis x (KM380), sLewis x (KM93), Lewis y (C14) and Tn, anti-MUC1 peptide core MAbs: C595, HMFG2 and SM3, anti-cytokeratins, anti-protoncogenes ErbB2 and ErbB3 (IgG) MAbs and also anti-CD34 and anti-CD45 MAbs. ELISA techniques were employed to study circulating MUC1 as well as free and complexed anti-MUC1 antibodies. Immunohistochemical results showed that carbohydrate antigenic expression increases in BM neoplastic cells compared to the original tumours. However, we were not able to demonstrate that a humoral immune response to MUC1 has been induced in these patients. Finally, the employed procedures allow the selective immortalisation of micrometastatic carcinoma cells since short-term cell lines were established.

[Effect of thermal treatments on the chemical characteristics of mora crab meat (Homalaspis plana)]. / Efecto de tratamientos térmicos sobre las características químicas de carne de jaiba mora (Homalaspis plana).

Marine species muscles present non-proteins nitrogenated compounds, used as quality index. They are total volatile basis (NBVT), trimethylamine oxide (TMAO) and trimethylamine (TMA). pH is considered too as a quality index. The aim of this work was to evaluate these parameters in a fresh and canned marine product from the V region, corresponding to mora crab (Homalaspis plana). Fresh pincer meat from mora crab was extracted and kept in ice until theits analysis and thermal process of the canned product. A 3(2) statistical design was applied, considering two variables with 3 levels: 15, 30 y 45 minutes time levels: 80 degrees, 100 degrees y 121 degrees C temperature levels. Nine conditions of time-temperature were obtained. The thermal treatment caused an increase in pH and BVT. The TMA was increased since reduction of TMAO.

Efecto de tratamientos térmicos sobre las características químicas de carne de jaiba mora (Homalaspis plana) / Effect of thermal treatments on the chemical characteristics of mora crab meat (Homalaspis plana)

Marine species muscles present non-proteins nitrogenated compounds, used as quality index. They are total volatile basis (NBVT), trimethylamine oxide (TMAO) and trimethylamine (TMA). pH is considered too as a quality index. The aim of this work was to evaluate these parameters in a fresh and canned marine product from the V region, corresponding to mora crab (Homalaspis plana). Fresh pincer meat from mora crab was extracted and kept in ice until theits analysis and thermal process of the canned product. A 3(2) statistical design was applied, considering two variables with 3 levels: 15, 30 y 45 minutes time levels: 80 degrees, 100 degrees y 121 degrees C temperature levels. Nine conditions of time-temperature were obtained. The thermal treatment caused an increase in pH and BVT. The TMA was increased since reduction of TMAO.

Detection of circulating mammary mucin (Muc1) and MUC1 immune complexes (Muc1-CIC) in healthy women.

There is convincing epidemiological evidence that multiparity provides protection against the development of breast cancer. In the present study we evaluated the levels of MUC1 and MUC1 circulating immune complexes (MUC1-CIC) in 135 serum samples obtained from healthy women. The study population included 13 women who had never been pregnant, 31 primiparous pregnant women, 36 multiparous pregnant women who had lactated, 5 multiparous pregnant women who had never lactated, 24 multiparous non-pregnant women who were lactating at the time of the study, 24 multiparous non-pregnant women who had lactated, and 2 multiparous non-pregnant women who had never lactated. The purpose of this work was to detect MUC1 variations during pregnancy and lactation as well as to study the possible induction of a humoral immune response against MUC1 in these conditions. We employed ELISA techniques to measure MUC1 (CASA test) and MUC1-CIC (IgM and IgG) using two anti-MUC1 monoclonal antibodies (MAbs): C595 and SM3. Statistical analysis was performed using the ANOVA test. The pooled results pertaining to pregnant versus non-pregnant women were compared and significant differences were observed in MUC1 and MUC1-CIC-lgM levels detected with both MAbs; the MUC1-CIC-lgG levels detected with C595 were increased in the pregnant group while the MUC1-CIC-lgG levels detected with SM3 did not show any significant differences. When the results were compared between lactating and non-lactating women, no significant differences were found. In conclusion, MUC1 and MUC1-CIC-lgM, detected with both MAbs, and MUC1-CIC-4gG levels detected with the MAb C595 are apparently induced by pregnancy.

Arthroscopic repair of acute traumatic anterior shoulder dislocation in young athletes.

PURPOSE: To compare the results of arthroscopic repair in acute anterior shoulder traumatic dislocation with those of nonoperative treatment. TYPE OF STUDY: A prospective nonrandomized study was performed. METHODS: Between August 1989 and April 1997, 46 patients were seen after a first episode of traumatic anterior shoulder dislocation. The average age was 21 years (range, 17 to 27 years). Most dislocations were in rugby players (36 patients). There were 18 patients treated by nonoperative methods and 28 patients treated by acute arthroscopic repair; 22 patients using transglenoid suture and 6 patients with bone anchor suture fixation. RESULTS: Of the patients treated nonoperatively, 94.5% suffered a redislocation between 4 and 18 months (average, 6 months). In the operative group, 96% of the patients (27) obtained excellent results according to the Rowe scale. Only 1 patient suffered a redislocation 1 year after surgery. Three different types of lesions were found during surgery: group I, capsular tear with no labrum lesion (4%); group II, capsular tear with partial labrum detachment (32%); and group III, capsular tear and full anterior labrum detachment (64%). The average follow-up was 67.4 months (range, 28 to 120). There were no surgical complications. CONCLUSIONS: The operative group obtained 96% excellent results, but the nonoperative group only obtained 5.5% excellent results, according to the Rowe scale. The nonoperative group showed a high incidence of redislocation (94.5%) compared with the operative group (4%). Based on the findings of this study, we recommend using an arthroscopic evaluation and repair after an initial anterior traumatic shoulder dislocation in young athletes.

Humoral immune response induced by the protein core of MUC1 mucin in pregnant and healthy women.

UNLABELLED: Serum levels of MUC1 and antibodies (Abs) against MUC1 (IgG and IgM-MUC1) were evaluated in healthy women related to pregnancy and lactation status. A total of 149 serum samples were obtained from: nulliparous, primiparous pregnant, multiparous pregnant that have lactated, multiparous pregnant without lactation, multiparous non-pregnant actual lactating, multiparous non-pregnant that have lactated and finally, multiparous non-pregnant women without lactation. In all assays, we included pre- and post-serum samples belonging to a breast cancer patient vaccinated with a MUC1 derived peptide. CASA test was employed to measure MUC1 while IgG- and IgM-MUC1 serum Abs were evaluated with an ELISA using a 100 mer peptide as catcher. In all groups, mean IgM levels were higher than IgG mean values; when samples were grouped in pregnants versus non-pregnants, a significant difference was detected with both Abs, being raised in non-pregnants. When samples were grouped in lactating versus non-lactating a significant difference was detected with IgG-MUC1, being raised in lactating women while no significant difference was found with IgM-MUC1. The evaluation of serum MUC1 levels confirmed previous results since a significant difference between pregnant versus non-pregnant groups was found while lactating versus non-lactating samples did not. CONCLUSIONS: (i) Increased MUC1 serum levels are apparently associated with pregnancy but not with lactation; (ii) MUC1 Abs are mainly associated with lactation and with non-pregnant status. These results may be considered a contribution on studies about protection against breast cancer induced by pregnancy and lactation.

[Effect of thermal treatment on the available lysine content of mora crab (Homalaspis plana) meat]. / Efecto de tratamientos térmicos sobre el contenido de lisina disponible en carne de jaiba mora (Homalaspis plana).

The effect of different high temperatures treatments on the available lysine content of mora crab meat, was studied. Fresh pincer meat from mora crab (from the V region) was extracted and kept in ice until the thermal process of the canned product. A 3(2) statistical design was applied, considering the following variables temperature (80 degrees C, 100 degrees C and 121 degrees C) and time (15, 30 and 45 minutes). Nine conditions temperature-time were obtained. Nutritional properties from available lysine were studied. A decrease from 8.33 (in raw meat) to 6.01 g/g protein in the most drastic thermal conditions, was observed. It ca be concluded that the content of available lysine in mora crab meat is more affected by time than by the temperature of the thermal treatments. Therefore the nutritive quality can be maintained applying high temperature and short time treatments.

Chilean legislation for the control of diseases of aquatic species.

The Political Constitution of Chile establishes the right of the people to live in an environment which is free from contamination. The State has a duty to uphold this right and to preserve nature. With the aim of reconciling this constitutional guarantee with the pursuit of profitable economic activities, the concept of 'sustainable development' has been promoted. This mechanism makes it possible to use natural resources and the environment in a rational, effective and efficient manner both at present and in the future, and led to the promulgation of the General Law of Fisheries and Aquaculture No. 18,892 of 6 September 1991 and its amendments. This law regulates national fisheries and aquaculture activities throughout the country, and contains eleven regulations applicable to aquaculture. These regulations form a complete procedure of action relevant to the importation of aquatic species, to applicants for aquaculture concessions and authorisations, to the operation of these concessions and authorisations, and to ocean ranching systems. Due to the complexity of these regulations, important rules are still under study, while the remaining texts come into force on 1 September 1996. Formulating these rules involves consideration of current national and international regulations, the experience of farmers, the results of technical and scientific research, and bibliographical data. These regulations will form the cornerstone of Children aquaculture policy in confronting the challenges of the next century.

Lateral discoid menisci in children.

In children, the snapping-knee syndrome is strongly suggestive of a diagnosis of discoid menisci; alternatively, the loss of physiological hyperextension of the knee suggests a diagnosis of a torn lateral discoid meniscus. We used arthroscopic meniscectomy to treat 19 lateral discoid menisci in 16 children (average age, 10.5 years). Excellent results were achieved in 18 of 19 cases, with disappearance of the snapping with knee flexion (maximum follow-up, 3 years).