Long-term evaluation of primary teeth molar pulpotomies with Biodentine and MTA: a CONSORT randomized clinical trial.

BACKGROUND: Calcium silicate-based materials have become widely used in recent years due to their positive effect on pulp cells, which stimulate tertiary dentin formation. AIM: The aim of the present study was to evaluate and compare clinically and radiographically the performance of MTA and Biodentine as pulp-dressing materials following pulpotomy in primary molars at 24-month follow-up. DESIGN: Molars from patients aged 4-9 years scheduled for pulpotomy were treated with either MTA or Biodentine followed by a stainless-steel crown. These molars were clinically and radiographically followed up at 6, 12, 18, and 24 months. Statistical analysis was performed to detect differences between the two groups. RESULTS: A total of 84 pulpotomies were performed obtaining a total success rate of 99.4 and 97.2% for Biodentine and MTA, respectively, at 24 months, showing no statistically significant differences between the two groups. CONCLUSION: 24-month follow-up showed that Biodentine and MTA have similar effectiveness.

Efectos de un programa de ejercicio neuromuscular en la condición física del adulto mayor de la comunidad: ensayo clínico aleatorizado en grupos paralelos y ciego doble / Effects of a neuromuscular exercise program on the physical condition of the older adult in the community: randomized clinical trial in parallel groups and double blind

OBJETIVO: Comparar los efectos de un programa de ejercicio neuromuscular (ENM) respecto a un programa de ejercicio municipal (EM) sobre la condición física en adultos mayores de la comunidad. Metodología: Ensayo clínico aleatorizado en grupos paralelos y ciego doble. Muestra constituida por 82 sujetos, edad X = 72 años (DE = +/-5 años) pertenecientes a la Casa del Adulto Mayor de la ciudad de Talca. La selección fue de manera no probabilística por conveniencia, la asignación a los grupos mediante aleatorización estratificada y el ocultamiento de la asignación por medio de sobres cerrados. El cegamiento doble fue para los participantes y los evaluadores. Ambos grupos realizaron ejercicio en paralelo durante seis meses, tres veces a la semana. Las comparaciones inter-grupo de la condición física fueron analizados por medio de ANOVA multivariante de un factor. En todos los casos se adoptó un p < 0.05 y se utilizó SPSS 25.Resultados: Los sujetos del grupo ENM lograron significativamente un mejor rendimiento en todas las variables de condición física respecto al grupo EM (p < 0.05). Conclusión: El ENM mejoró significativamente la condición física de adultos mayores de la comunidad respecto a la aplicación de un programa de ejercicio municipal.

OBJECTIVE: To compare the effects of a neuromuscular exercise program (ENM) with respect to a municipal exercise program (EM) on the physical condition of older adults in the community. Methodology: Double blind, parallel group randomized clinical trial. Sample made up of 82 subjects, age X = 72 years (SD = +/- 5 years) belonging to the House for the Elderly in the city of Talca. Selection was non-probabilistic for convenience, group assignment by stratified randomization, and allocation concealment by sealed envelopes. Double blinding was for participants and assessors. Both groups exercised in parallel for 6 months, 3 times a week. The inter-group comparisons of physical condition were by means of one-way multivariate ANOVA. In all cases a p <0.05 was adopted and SPSS 25 was used. Results: The subjects of the ENM group achieved significantly better performance in all the physical condition variables compared to the EM group (p <0.05). Conclusion: The ENM significantly improved the physical condition of older adults in the community with respect to the application of a municipal exercise program.

Calidad de la educación sexual recibida en el contexto escolar y su asociación a conductas sexuales en adolescentes chilenos, según datos VIII Encuesta Nacional de la Juventud / Quality of sexual education received in the school context and its association with sexual behaviors in Chilean adolescents, according to data from the VIII National Youth Survey

OBJETIVO: Evaluar la calidad de la educación sexual recibida en el contexto escolar y su asociación a conductas sexuales en adolescentes chilenos. MÉTODOS: Estudio transversal y analítico. La muestra correspondió a adolescentes de ambos sexos de 15 a 19 años, se obtuvo de una base de datos con representación nacional. Se evaluó la calidad de la educación sexual recibida en el contexto escolar como buena, regular y mala. Se realizó análisis descriptivo y de asociación entre las variables. Se calculó OR (Odds ratio) y RRR (Relative-risk ratios) ajustado por sexo, edad en años cumplidos, grupo socioeconómico y zona. RESULTADOS: La muestra correspondió a 1.049 adolescentes sexualmente activos, de estos el 21,83% correspondió al grupo con evaluación mala, 44,16% con evaluación regular y 34,01% con evaluación buena. La razón entre adolescentes que no usaron condón en la última relación sexual versus usaron condón, es 1,64 veces mayor en los adolescentes con evaluación mala de la educación sexual recibida (OR: 1,64; IC95%: 1,01 - 2,68). La razón entre adolescentes que tuvieron dos o más parejas sexuales en el último año versus ninguna o una, es 2,15 veces mayor en los adolescentes con evaluación mala de la educación sexual recibida (OR: 2,15; IC95%: 1,30 - 3,56). CONCLUSIÓN: Los participantes de este estudio evalúan la educación sexual recibida mayoritariamente como regular, existiendo una asociación con las conductas sexuales. Es importante realizar estudios en profundidad que permitan conocer por qué los adolescentes evalúan sólo como regular la educación sexual recibida en el sistema escolar.

OBJECTIVE: To evaluate the quality of the sexual education received in the school context and its association with sexual behaviors in Chilean adolescents. METHODS: Cross-sectional and analytical study. The sample corresponded to adolescents of both sexes from 15 to 19 years old, it was obtained from a database with national representation. The quality of the sexual education received in the school context was evaluated as good, regular and bad. Descriptive and association analysis were performed between the variables. OR (Odds ratio) and RRR (Relative-risk ratios) adjusted for sex, age in completed years, socioeconomic group and area were calculated. RESULTS: The sample corresponded to 1,049 sexually active adolescents, of these, 21.83% corresponded to the group with bad "Evaluation", 44.16% with regular "Evaluation" and 34.01% with good "Evaluation". The ratio among adolescents who did not use a condom in the last sexual relationship versus those who used a condom is 1.64 times higher in adolescents with a poor evaluation of sexual education received (OR: 1.64; 95% CI: 1.01 - 2.68). The ratio among adolescents who had two or more sexual partners in the last year versus those who had neither or one, is 2.15 times higher in adolescents with poor evaluation of sexual education received (OR: 2.15; 95% CI: 1.30 - 3.56). CONCLUSION: The participants of this study evaluate the sexual education received mostly as regular, with an association with sexual behaviors. It is important to conduct in-depth studies that allow us to know why adolescents evaluate the sexual education received in the school system as regular only.

In vitro and in vivo development of domestic cat embryos generated by in vitro fertilization after eCG priming and oocyte in vitro maturation.

The objective of this study was to evaluate, in the domestic cat, the effect of ovarian stimulation with eCG prior to oocyte in vitro maturation (priming) on in vitro and in vivo development after in vitro fertilization (IVF). For this purpose, oocyte donors were either 1) treated with a single dose of 200 IU eCG four days before oocyte recovery (eCG group), or, 2) given no treatment before oocyte recovery (control group). Ovaries of both groups were collected by ovariohysterectomy (OVH) and cumulus-oocyte complexes (COCs) were recovered by slicing. Immature COCs from both groups were matured in vitro (IVM) for 26-28 h. IVF was done with refrigerated epididymal sperm. After 24 h co-incubation, presumptive zygotes were cultured in vitro for eight days. The rates of cleavage, morulae, blastocyst development and hatching were estimated. Some blastocysts were stained for total cell counting and others were used for gene expression analysis of pluripotency (OCT4, SOX2 and NANOG) and differentiation markers (CDX2 and GATA6). Additionally, to evaluate in vivo development, embryos from the eCG group were transferred at Day 5 and Days 7 or 8 of IVC to synchronized cat recipients. The results showed that, eCG priming increased significantly the rate of blastocyst development as compared to the control group (37.9 and 25.6%, respectively) (P < 0.05). No differences were observed in total cell number of blastocysts and hatching blastocysts (mean ± SD) between the eCG and control groups (420.6 ± 193.6 and 347.0 ± 237.1, respectively) (P > 0.05). In the gene expression analysis, blastocysts generated in the eCG group had higher expression of OCT4 than blastocysts from the control group (P < 0.05). However, no significant differences were observed in the relative expression of SOX2, NANOG, CDX2 and GATA6 (P > 0.05). Additionally, six embryo transfer (ET) procedures were done, three with Day 5 embryos and three with Day 7 or 8 embryos. Recipients from both ET groups delivered live kittens. The total pregnancy rate was 4/6 (67%), meanwhile the live birth rate was 2/6 (33%). In conclusion, eCG priming improved the rate of blastocyst development in vitro and increased relative expression of OCT4. These results demonstrate that eCG priming of oocytes donors before IVM improves oocyte competence, enhance in vitro embryo development and allows live births of healthy offspring after ET.

Salud sexual y salud reproductiva en mujeres jóvenes con consumo de sustancias en Chile según datos VIII encuesta nacional de la juventud / Sexual health and reproductive health in young women with substance consumption in Chile according to data VIII national youth survey

OBJETIVOS: Determinar la asociación entre el consumo de sustancias y características de salud sexual y reproductiva de mujeres jóvenes en Chile. MÉTODOS: Estudio transversal analítico en mujeres entre 15 a 24 años. Se formaron 3 grupos: Sin consumo de sustancias (SCS), Consumo no problemático de sustancias (CNPS) y Consumo problemático de sustancias (CPS). Se realizó análisis descriptivo y de asociación entre las variables. Se ajustaron modelos de regresión logística múltiple y ordinal múltiple. RESULTADOS: Muestra de 2.589 jóvenes, el 37,8% correspondió al grupo SCS, 46,3% al grupo CSNP y 15,9% al grupo CPS. En promedio la edad de inicio de actividad sexual fue menor en el grupo CPS (15,9 años p = 0,001) mostrando mayor porcentajes de sexo oral, anal, no uno de condón, relaciones sexuales con parejas menos estables, mayor número de parejas sexuales y violencia en la pareja, (p =0,001). En este grupo se incrementa 5,84 el riesgo de tener la última relación sexual con pareja menos estable (IC95%: 3,90 - 12,01) y 8,35 veces el riesgo de tener 2 o más parejas sexuales (últimos 12 meses) (IC95%: 5,35 - 16,34). En el grupo CNPS se incrementa 1,11 veces el riesgo de tener la última relación sexual con pareja menos estable (IC95%: 1,43 - 3,12), 1,01 veces tener 2 y más parejas sexuales (últimos 12 meses) (IC95%: 1,20 - 3,36). CONCLUSIONES: Reconociéndose que la sexualidad es multifactorial, las mujeres con consumo de sustancias viven situaciones que facilitan riesgos para su salud sexual. El consumo en mujeres jóvenes es un problema de salud pública que presenta desafíos para su abordaje.

OBJECTIVES: To determine the association between substance use and characteristics of sexual and reproductive health of young women in Chile. METHODS: Analytical cross-sectional study in women between 15 and 24 years. Three groups were formed: No substance use (SCS), Non-problematic substance use (CNPS) and Problem substance use (CPS). Descriptive and association analysis was performed between the variables. Multiple logistic regression and multiple ordinal models were adjusted. RESULTS: Sample of 2,589 young people, 37.8% corresponded to the SCS group, 46.3% to the CSNP group and 15.9% to the CPS group. On average the age of onset of sexual activity was lower in the CPS group (15.9 years p = 0.001) showing higher percentages of oral, anal sex, not a condom, sex with less stable partners, greater number of sexual partners and violence in the couple, (p = 0.001). In this group the risk of having the last sexual relationship with a less stable partner is increased 5.84 (95% CI: 3.90 - 12.01) and 8.35 times the risk of having 2 or more sexual partners (last 12 months) (95% CI: 5.35-16.34). In the CNPS group the risk of having the last sexual relationship with a less stable partner is increased 1.11 times (95% CI: 1.43 - 3.12), 1.01 times having 2 and more sexual partners (last 12 months) (95% CI: 1.20-3.36). CONCLUSIONS: Recognizing that sexuality is multifactorial, women with substance use experience situations that facilitate risks to their sexual health. Consumption in young women is a public health problem that presents challenges for its approach.

Alteraciones tiroideas: ¿Predictores de mortalidad en Cuidados Intensivos? / Thyroid disorders: Predictors of mortality in Intensive Care?

Introducción: El objetivo del estudio es describir y analizar la función tiroidea en el paciente crítico. Describir si algún patrón se asocia a una mayor tasa de mortalidad. Métodos: Se analizó a todos los pacientes ingresados en nuestra Unidad de Cuidados Intensivos, entre enero de 2015 y agosto de 2016, y que permanecieron allí, al menos, siete días. Resultados: Se incluyeron 242 pacientes. Se hallaron diferencias significativas en los valores medios de los índices de gravedad entre los no supervivientes y los supervivientes: SOFA (10,45 vs. 7,9); APACHE II (24,42 vs. 20,71); SAPS II (63,14 vs. 50,69). Se encontraron diferencias estadísticamente significativas en los valores medios de T3 en el grupo de supervivientes y no supervivientes: 1,5 pg/ml vs. 1,15 pg/ml (p <0,001; IC95% 0,224 ± 0,487). No hubo diferencias estadísticamente significativas en los valores medios de T4 y TSH. Al realizar el subanálisis según grupo de ingreso, no se hallaron diferencias estadísticamente significativas entre las cifras de TSH, T3 o T4. Sí hubo diferencias significativas en los valores medios de T3 en la mayor parte de los subgrupos. Conclusiones: Se hallaron diferencias estadísticamente significativas entre los valores medios de T3 en el grupo de supervivientes y no supervivientes. Los valores de T3 parecen asociarse a la mortalidad. Su descenso no parece asociarse a la enfermedad subyacente, sino a su gravedad.(AU)

Introduction: The aim of this study is to describe and analyse the thyroid function in the critically ill patient, and to describe if any pattern is associated with a higher mortality rate. Methods: Patients admitted to the Intensive Care Unit, between January 2015 and August 2016, with a stay of seven days or more, were enrolled. Results: Two hundred and forty-two patients were included. Significant differences were observed in the severity scores related to mortality during their stay in the Intensive Care Unit (no survivors vs. survivors): SOFA (10.45 vs. 7.9); APACHE II (24.42 vs. 20.71); SAPS II (63.14 vs. 50.69). Statistically significant differences were observed in the mean values of T3 between survivors and non-survivors: 1.5 pg/ml vs. 1.15 pg/ml (p <0.001; CI95% 0.224 ± 0.487). There were no statistically significant differences in the mean values of T4 and TSH. After subgroup analysis according to the admission group, no significant differences among the TSH, T3 and T4 values were detected. However a statistically difference in T3 average value was found in most subgroups. Conclusions: A statistically significant difference was found in mean T3 values in survivors and non-survivors. T3 concentration appears to be associated with mortality. Decrease of this thyroid hormone does not seem to associate with the underlying disease, but with its severity.(AU)

Efficacy of an extravascular lung water-driven negative fluid balance protocol.

OBJECTIVE: To analyze the efficacy of negative fluid balance in hypoxemic patients with an elevated extravascular lung water index (EVLWI). DESIGN: A retrospective observational study was made. SETTING: Intensive Care Unit of Virgen de las Nieves Hospital (Spain). PARTICIPANTS: Forty-four patients participated in the study. INTERVENTIONS: We analyzed our database of hypoxemic patients covering a period of 11 consecutive months. We included all hemodynamically stable and hypoxemic patients with EVLWI>9ml/kg. The protocol dictates a negative fluid balance between 500 and 1500ml/day. We analyzed the impact of this negative fluid balance strategy upon pulmonary, hemodynamic, and renal function. MAIN VARIABLES OF INTEREST: Demographic data, severity scores, clinical, hemodynamic, pulmonary, metabolic and renal function data. RESULTS: Thirty-three patients achieved negative fluid balance (NFB group) and 11 had a positive fluid balance (PFB group). In the former group, PaO2/FiO2 improved from 145 (IQR 106, 200) to 210mmHg (IQR 164, 248) (p<0.001), and EVLWI decreased from 14 (11, 18) to 10ml/kg (8, 14) (p<0.001). In the PFB group, EVLWI also decreased from 11 (10, 14) to 10ml/kg (8, 14) at the end of the protocol (p=0.004). For these patients there were no changes in oxygenation, with a PaO2/FiO2 of 216mmHg (IQR 137, 260) at the beginning versus 205mmHg (IQR 99,257) at the end of the study (p=0.08). CONCLUSION: Three out of four hypoxic patients with elevated EVLWI tolerated the NFB protocol. In these subjects, the improvement of various analyzed physiological parameters was greater and faster than in those unable to complete the protocol. Patients who did not tolerate the protocol were usually in more severe condition, though a larger sample would be needed to detect specific characteristics of this group.

High expression of gabarapl1 is associated with a better outcome for patients with lymph node-positive breast cancer.

BACKGROUND: This study evaluates the relation of the early oestrogen-regulated gene gabarapl1 to cellular growth and its prognostic significance in breast adenocarcinoma. METHODS: First, the relation between GABARAPL1 expression and MCF-7 growth rate was analysed. Thereafter, by performing macroarray and reverse transcriptase quantitative-polymerase chain reaction (RT-qPCR) experiments, gabarapl1 expression was quantified in several histological breast tumour types and in a retrospective cohort of 265 breast cancers. RESULTS: GABARAPL1 overexpression inhibited MCF-7 growth rate and gabarapl1 expression was downregulated in breast tumours. Gabarapl1 mRNA levels were found to be significantly lower in tumours presenting a high histological grade, with a lymph node-positive (pN+) and oestrogen and/or progesterone receptor-negative status. In univariate analysis, high gabarapl1 levels were associated with a lower risk of metastasis in all patients (hazard ratio (HR) 4.96), as well as in pN+ patients (HR 14.96). In multivariate analysis, gabarapl1 expression remained significant in all patients (HR 3.63), as well as in pN+ patients (HR 5.65). In univariate or multivariate analysis, gabarapl1 expression did not disclose any difference in metastasis risk in lymph node-negative patients. CONCLUSIONS: Our data show for the first time that the level of gabarapl1 mRNA expression in breast tumours is a good indicator of the risk of recurrence, specifically in pN+ patients.

Instilación intratraqueal de elastasa en la rata: cambios electroforéticos de la alfa1-AT-antitripsina en el lavado broncoalveolar / Intratracheal instillation of elastase in the rat: electrophoretical changes of alphal-antitrypsin in bronchoalveolar lavage fluid

Introduction: Endogenous alphal-antitrypsin alpha is the main inhibitor of the intratracheally instilled elastase in experimental animals. Objective: To evaluate by electrophoresis and immunodetection using western blot analysis, the different forms of alpha1-AT in bronchoalveolar lavage fluid (BALF) of Sprague Dawley rats after intratracheal instillation of elastase, with the hypothesis that the previously observed increment in antielastase activity is due to high levels of active alpha1-AT. Results: In the first hours after elastase instillation the concentration of alpha1-AT increases more than seven times due to an increase in alveolar-capillary permeability. Alpha 1-AT in BAIF is found as the native protein (~ 52 kDa), as complexes of different molecular sizes (> 75 kDa and > 100 kDa) and as a proteolytic product (< 40 kDa). Conclusion: In spite of a high proportion of alpha1-AT in the inactive form as part of different complexes, the increase in alveolar-capillary permeability after elastase treatment contributes to maintain high levels of active alpha. These results could be of importance in other inflammatory lung processes.

Introducción: la antiproteasa alfa 1-antitripsina alfa constituye el principal inhibidor endógeno de la elastasa instilada por vía intratraqueal en modelos experimentales. Objetivo: Evaluar mediante electroforesis e inmunodetección por western blot, las distintas formas en que se encuentra la alfa1-AT en el lavado broncoalveolar (IBA) de ratas Sprague Dawley después de la instilación de elastasa, con la hipótesis de que el aumento en la actividad antielastasa previamente encontrada se acompaña de niveles altos de alfa1-AT activa. Resultados: En las primeras horas post-elastasa la concentración de alfa1-AT en el IBA aumenta más de 7 veces, debido al aumento de la permeabilidad alvéolo-capilar, encontrándose tanto como proteína nativa (~ 52 kDa), como parte de complejos de mayor tamaño (> 75 kDa y > 100 kDa) y como producto de proteólisis (< 40 kDa). Conclusión: A pesar de existir una alta proporción de alfa1-AT inactiva formando complejos, el aumento de la permeabilidad alvéolo-capilar contribuye a mantener niveles altos de alfa1-AT activa. Estos resultados podrían ser extrapolables a distintos procesos inflamatorios pulmonares.

The helminth community of the wood mouse, Apodemus sylvaticus, in the Sierra Espuna, Murcia, Spain.

The helminth community of the wood mouse, Apodemus sylvaticus, in the Sierra Espuna was characterized after a complete analysis of its helminth community component and infracommunity structure relative to host age, sex and year of capture. The helminth community comprised 13 species: one trematode, four cestodes and eight nematodes. The cestode Pseudocatenotaenia matovi and the nematode Syphacia frederici were the most prevalent and abundant helminth species, respectively. Sixty four percent of mice analysed presented helminths with a direct cycle and 42% presented helminths with an indirect cycle. The helminth community presents a low diversity with infracommunities usually made up of only one or two helminth species. Host age and year of capture seem to play a major role in determining species richness and helminth diversity, but not in determining the abundance of helminths. Host sex does not seem to affect the infection rate nor the diversity. Further studies on more samples of wood mice and other small mammal species in this regional park are needed to explore any possible interactions between helminth communities in the host populations.

[Bilateral lagophthalmos in lepromatous leprosy. Case report]. / Lagoftalmos bilateral en paciente con lepra lepromatosa. Caso clínico.

CASE REPORT: A case of bilateral facial palsy with paralytic ectropion, lagophthalmos and corneal damage secondary to corneal exposure in a long-standing patient with lepromatous leprosy is presented. Correction of paralytic ectropion was performed by medial cantoplasty, tarsal strip and Medpor lower eyelid spacer implantation. Lagophthalmos was corrected by gold weight implant in the upper tarsus. DISCUSSION: Ocular findings in leprosy appear in 72% of patients. Facial nerve palsy occurs in 3-19.8%, being bilateral in 5%. In long standing cases with corneal complications secondary to exposure, surgical treatment is required.

Quiste hidatídico de la lengua / Hydatid cyst of the tongue

La lengua es una rara localización de la hidatidosis. Se reporta el caso de una paciente de 3 años con enfermedad hidatídica de la lengua demostrada histológicamente. El quiste de la lengua fue extirpado quirúrgicamente. La paciente tenía además hidatidosis hepática y pulmonar que fueron tratadas exitosamente con Albendazol

Differential regulation of normal and tumoral breast epithelial cell growth by fibroblasts and 1,25-dihydroxyvitamin D3.

Mesenchymal-epithelial interactions are of paramount importance during normal and tumoral breast developments. We have investigated the paracrine growth regulation of normal and tumoral breast epithelial cells by fibroblasts derived from normal or pathological breast tissues. In some cases, breast cancer MCF-7 cells or normal epithelial cells in primary culture were cocultured with fibroblasts in a Transwell system allowing diffusible factor exchanges. Alternatively, conditioned medium produced by fibroblast cultures was added to epithelial cell cultures. Fibroblasts were shown to stimulate the proliferation of normal and carcinoma cells through paracrine mechanisms. However, the paracrine exchanges appeared to be different in normal versus tumoral breast epithelial cell growth regulation. Moreover, vitamin D-related compounds that have been proposed as anti-tumoral drugs were studied for their ability to affect normal and tumoral mammary epithelial cell proliferation and to interfere with the growth-regulatory activity of fibroblasts. Whereas vitamin D compounds inhibited MCF-7 cell growth, they led to a marked stimulation of the proliferation of normal mammary epithelial cells. Moreover, it was shown that the vitamin D analog EB 1089 can block the mitogenic effect of fibroblast-conditioned medium on tumoral but not normal breast epithelial cells. The differential effects of vitamin D compounds on cell proliferation provide further data in favor of the different behaviours of normal and tumoral mammary epithelial cells. The potential therapeutic use of vitamin D derivatives in the treatment of breast cancer is supported by these results but their growth-stimulatory properties on normal epithelial cells cannot be overlooked.

[Long-term prognostic role of steroid receptors in cancer of the breast]. / Rôle pronostique à long terme des récepteurs stéroïdiens dans le cancer du sein.

We screened for the prognostic value of estrogen receptor (ER) and progesterone receptor (PR) through a multicentric study of 2,257 operable breast cancer patients who did not received adjuvant therapy. Three hundred and seven local-regional recurrences, 105 metachronous contralateral breast cancer, 589 metastases and 537 deaths from cancer had been diagnosed with a median follow-up of 8.5 years. A total of 69% of the tumors were ER positive and 54% PR positive. For statistical analysis, 1,665 patients were studied because of complete clinical and biological data. In univariate analysis, ER and PR status were of prognostic value for the metastases-free interval (MFI) and the overall survival (OS). In multivariate analysis (Cox proportional hazard model), only the ER status showed a significant difference between positive and negative groups regarding the MFI and OS. By using Cox regression model with time-dependent covariates, we show that the predictive value of ER status of the primary tumor decreases by approximately 20% per year, losing its significance after 8 years of follow-up. These results show that ER and PR status have a relatively limited predictive value and their major interest remain in the domain of therapeutic decision.

Tissue extraction procedures for investigation of urokinase plasminogen activator (uPA) and its inhibitors PAI-1 and PAI-2 in human breast carcinomas.

Urokinase type plasminogen activator (uPA) and its inhibitors, plasminogen activator inhibitor type I (PAI-1) and type II (PAI-2), are supposed to be involved in the expression of the invasive and metastatic phenotype of cancer cells. However, clinical investigations on the prognostic significance of their levels in tumor tissue are difficult to realize because of the absence of a convenient method of measurement of these parameters. The aim of the present investigation was to set up a method allowing the measurement of these enzymes and of sex steroid receptor status in appropriate subcellular fraction(s) in conditions easily reproducible in routine. We found that a tissue homogenate prepared according to the method recommended [5] for current measurement of sex steroid receptors is appropriate for further distinct preparations. One aliquot is used for cytosol preparation; another can be treated by 2% Triton X-100 (vol/vol) and provide an extract containing the totality of uPA and PAI-1. The advantage of this procedure is that appropriate subcellular fractions can be derived from a unique homogenization step. Total uPA and PAI-1 are measured in a Triton extract with good performance as compared to previous investigations [4]. PAI-2 is measured in the same cytosol fraction used for sex steroid receptors and other parameters. Because of its simplicity and its high reliability, this method could be a useful tool in the investigation of uPA family proteases and analysis of their prognostic significance in early breast tumors.

Positive regulation of normal and tumoral mammary epithelial cell proliferation by fibroblasts in coculture.

In the mammary gland, mesenchymal-epithelial interactions are of paramount importance during normal and tumoral developments. We have studied the paracrine growth regulation of a variety of breast epithelial cells in coculture with normal or pathological breast fibroblasts. Two models of coculture were used in which the two cell types were seeded and grown, either together in microchamber slides or separated by a microporous membrane. Under these two conditions, all fibroblasts were shown to stimulate the proliferation of the hormono-responsive breast carcinoma MCF-7 cell line, suggesting that cell contacts were not indispensable for the paracrine stimulation of MCF-7 cell growth by fibroblasts. Moreover, in the Transwell coculture system, the proliferation of a variety of other breast carcinoma cells (MDA-MB231, T47D, and BT-20) was also stimulated by fibroblasts. However, the amplitude of the proliferative response seemed to be dependent on the carcinoma cell line considered. Moreover, the proliferative response of normal mammary epithelial cells to the presence of fibroblasts was shown to be significantly higher than the tumor cell response. The nature of the tissue of fibroblast origin, normal or pathological, did not influence the growth response of the epithelial cells. In this study, we thus demonstrate that fibroblasts are able to stimulate the proliferation of normal and carcinoma cells through paracrine exchange mechanisms. We also conclude that the target epithelial cell phenotype will essentially determine the extent of the proliferative response.

Epidermal growth factor and transforming growth factor alpha mRNA expression in human breast cancer biopsies; analysis in relation to estradiol, progesterone and EGF receptor content.

Among the peptide growth factors active in breast glandular cell proliferation epidermal growth factor (EGF) and transforming growth factor alpha (TGF alpha) are thought to play a major role in tumour development. They operate through binding to and activation of a common membrane receptor, defined as EGF-R. Their production is modulated by hormones and local growth factors. After it was shown by previous investigation in this laboratory that EGF-R could be detected in 90% of the tumours, but was masked by endogenous ligand in 36% of them, the question was raised as to the level of the ligand's expression in tumour tissue biopsies. Therefore, we investigated the expression of EGF and TGF alpha mRNA in 146 breast cancer biopsies by slot blot analysis using specific 32P-labelled probes. The data were correlated with sex steroids and EGF receptor content. Our results showed that EGF and TGF alpha coexisted in all tumour samples, and that their level of mRNA expression was similar in half of the tumours. Northern blot and polymerase chain reaction (PCR) analysis validated these findings. A significant direct correlation was found between the level of TGF alpha/EGF mRNA expression and the ER/progesterone receptor (PGR) content. TGF alpha and EGF mRNA levels were significantly higher in ER+ (P = 0.0015 and P = 0.0001, respectively) and in PGR+ tumours (P < 0.005 and P = 0.0001) than in their negative counterparts. Moreover, TGF alpha mRNA expression negatively correlated with the number of EGF-R binding sites measured by the standard method (P = 0.02), and it was significantly related to the number of sites occupied by endogenous ligand. In conclusion, it was shown that TGF alpha and EGF mRNA were coexpressed in all the tumour biopsies tested and that their level was higher in the hormone receptor positive than in negative samples. The correlation between the presence of ER/PGR sites, high level of TGF alpha/EGF mRNA and EGF-R occupancy by endogenous ligand is in favour of ER mediated control of TGF alpha and EGF production.

Rat stromelysin 3: cDNA cloning from healing skin wound, activation by furin and expression in rat tissues.

From a rat skin wound healing cDNA library, a clone encoding stromelysin 3 (ST3) was isolated. The predicted rat ST3 has 491 amino acids, and shows 83, 95 and 58% homology with human, mouse and Xenopus ST3, respectively. COS-1 cells transfected with this rat ST3 cDNA produced the ST3 proform, which could be converted to the mature ST3 form by co-transfection with rat furin cDNA. In addition to healing skin, the rat ST3 gene was found to be strongly expressed in normal adult uterus and ovary, and at lower levels in chemically-induced mammary tumors.

Comparison of a new microplate oestrogen receptor (ER) enzyme immunoassay with other ER detection methods.

In a study involving 50 breast cancer tumours, we compared two oestrogen receptor (ER) detection methods developed by us--a microplate immunoenzymometric assay (EIA96) and an immunohistochemistry kit (HistoCIS-ER)--with the radioligand assay (RLA), the Abbott immunoenzymometric assay ER-EIA and the reverse transcriptase polymerase chain reaction technique (RT-PCR). Among the three ER protein cytosolic assays (EIA96, ER-EIA and RLA), the two EIAs showed the best agreement (y = 1.086x - 7.840; r2 = 0.876). At the calculated optimal cut-off values (8 and 14 fmol mg(-1) protein for EIA96 and RLA respectively), EIA96 was more sensitive than RLA (0.94 for EIA96, 0.88 for RLA), but slightly less specific (0.82 for EIA96, 0.94 for RLA). The Cox logistical regression model applied to EIA96, RLA and RT-PCR showed that EIA96 discriminated the best between ER-EIA+ and ER-EIA- samples. The RT-PCR technique and HistoCIS-ER both had a positivity-negativity concordance of 86% with EIA96.