Total circulating microparticle levels are increased in patients with deep infiltrating endometriosis.

STUDY QUESTION: Are the levels of total circulating cell-derived microparticles (cMPs) and circulating tissue factor-containing microparticles (cMP-TF) increased in patients with endometriosis? SUMMARY ANSWER: The levels of total cMP, but not cMP-TF, were higher in patients with endometriosis, and these were attributed to higher levels in patients with deep infiltrating endometriosis (DIE). WHAT IS KNOWN ALREADY: Previous studies have reported elevated levels of total cMP in inflammatory conditions as well as higher levels of other inflammatory biomarkers in endometriosis. Increased expression of tissue factor (a transmembrane receptor for Factor VII/VIIa) in eutopic and ectopic endometrium from patients with endometriosis has been described. There is no previous data regarding total cMP and cMP-TF levels in patients with endometriosis. STUDY DESIGN, SIZE, DURATION: A prospective case-control study including two groups of patients was carried out. The E group included 65 patients with surgically confirmed endometriosis (37 with DIE lesions) and the C group comprises 33 women without surgical findings of any form of endometriosis. Patients and controls were recruited during the same 10-month period. Controls were the next patient without endometriosis undergoing surgery, after including two patients with endometriosis. PARTICIPANTS/MATERIALS, SETTING, METHODS: Venous blood samples for total cMP and cMP-TF determinations were obtained at the time of surgery, before anesthesia at a tertiary care center. To assess total cMP, an ELISA functional assay was used and cMP-TF activity in plasma was measured using an ELISA kit. MAIN RESULTS AND THE ROLE OF CHANCE: Total cMP levels in plasma were higher in the E group compared with the C group (P < 0.0001). The subanalysis of endometriosis patients with DIE or with ovarian endometriomas without DIE showed that total cMP levels were higher in the DIE group (P = 0.001). There were no statistically significant differences in cMP-TF levels among the groups analyzed. LIMITATIONS, REASONS FOR CAUTION: This is a preliminary study in which the sample size was arbitrarily decided, albeit in keeping with previous studies analyzing cMP in other inflammatory diseases and other biomarkers in endometriosis. The control group included patients with other pathologies as well as healthy controls, and blood samples were taken at different phases of the cycle. WIDER IMPLICATIONS OF THE FINDINGS: Elevated total cMP levels in DIE patients may reflect an inflammatory and/or procoagulant systemic status in these patients. Further studies are warranted to confirm our findings and to assess the role of cMP levels in the pathophysiology of DIE. STUDY FUNDING/COMPETING INTERESTS: This study was supported in part by a grant from FIS-PI11/01560 and FIS-PI11/00977 within the 'Plan Nacional de I + D + I' and co-funded by the 'ISCIII-Subdirección General de Evaluación' and 'Fondo Europeo de Desarrollo Regional (FEDER)' and by the grant 'Premi Fi de Residència Emili Letang 2015' from the Hospital Clínic of Barcelona. The authors have no competing interests to disclose.

Long lasting behavioural effects on cuprizone fed mice after neurotoxicant withdrawal.

Destruction of the myelin sheath in the central nervous system (CNS) is prominent in many clinico-pathologic conditions. Among animal models that reproduce the pathological features of de- and remyelination processes, the mouse model of cuprizone administration is widely used. Both hyperactivity and motor impairment have been reported upon cuprizone exposure. The aim of the present study was to assess behaviour in mice after CPZ withdrawal.To summarize, animals showed hypo-activity and deficits in motor coordination when they were subjected to acute demyelinating insult while minor exploratory activity, impairment in motor coordination and lower anxiety levels emerged when remyelination was reached following cuprizone withdrawal. A recovery period of 6 weeks after removal of CPZ was not accompanied by a similar return of normal activity indicating long lasting behavioural effects caused by this neurotoxicant. Specifically, the recovery group showed impairments in neurological functions involved in sensorimotor, neuromuscular, motor coordination and the capacity to cope with a stress-inducing event.

Inhibition of serotonin receptor type 1 in acute myeloid leukemia impairs leukemia stem cell functionality: a promising novel therapeutic target.

Acute myeloid leukemia (AML) is a clinically and molecularly heterogeneous neoplasia with poor outcome, organized as a hierarchy initiated and maintained by a sub-population with differentiation and self-renewal capacities called leukemia stem cells (LSCs). Although currently used chemotherapy is capable of initially reducing the tumor burden producing a complete remission, most patients will ultimately relapse and will succumb to their disease. As such, new therapeutic strategies are needed. AML cells differentially expressed serotonin receptor type 1 (HTR1) compared with healthy blood cells and the most primitive hematopoietic fraction; in fact, HTR1B expression on AML patient samples correlated with clinical outcome. Inhibition of HTR1s activated the apoptosis program, induced differentiation and reduced the clonogenic capacity, while minimal effect was observed on healthy blood cells. In vivo regeneration capacity of primary AML samples was disrupted upon inhibition of HTR1. The self-renewal capacity remaining in AML cells upon in vivo treatment was severely reduced as demonstrated by serial transplantation. Thus, treatment with HTR1 antagonists showed antileukemia effect, especially anti-LSC activity while sparing healthy blood cells. Our results highlight the importance of HTR1 in leukemogenesis and LSC survival and identify this receptor family as a new target for therapy in AML with prognostic value.

Breast cancer response to adjuvant chemotherapy in correlation with erbB2 and p53 expression.

The expression of the erbB2 and p53 proteins was examined by immunohistochemistry in a series of 81 cases of primary breast carcinomas treated with an adjuvant chemotherapy with a median follow-up of 5 years. Expression of erbB2 was present in 19 cases (23%), 50% of them with more than 70% of positive tumoral cells. Expression of p53 was present in 32 cases (39%), 50% of them with more than 30% of positive tumoral cells; coexpression of erbB2 and p53 was observed in 11 cases (13.5%). Disease-free and overall survival of the patients were correlated with the number of positive lymph nodes (p = 0.08, p = 0.015). No significant trend was observed in response to adjuvant chemotherapy in correlation with erbB2 or p53 expression or both, whatever the level of expression.

Embryonic development of pineal gland vesicles: a morphological and morphometrical study in chick embryos.

Pineal cell aggregates in 5, 10 and 15 day-old chick embryos have been studied. Cell aggregates were classified into rosettes or vesicles and spheroid and ellipsoid vesicles distinguished. The number of pineal vesicles per unit of surface (vesicle density) was determined in three pineal portions: apical, anterior and posterior. By day 5, only cellular rosettes were found, mainly in the apical portion. After 10 and 15 days, the presence of rosettes was occasional. The posterior wall showed only small spheroid vesicles, while in the apical and anterior areas ellipsoid vesicles were also observed. Moreover, the spheroid/ellipsoid vesicle ratio increased from the 10th to the 15th day of incubation. The vesicle density decreased between the 10th and 15th day because of the increase in both vesicle and pineal size, without changes in the total number of vesicles. The results suggest that changes in vesicle morphology and density could be related to the functional activity of the pineal gland during embryonic development.

[Human breast cancer embedded in paraffin, fixed in Bouin's fluid. Immunohistochemical analysis of estrogen and progesterone receptors]. / Carcinomes mammaires humains inclus en paraffine, fixés dans le liquide de Bouin. Analyse immunohistochimique des récepteurs d'estrogène et de progestérone.

A new immunohistochemical method using a monoclonal estrogen receptor (ER) antibody and a progesterone receptor (PR) antibody has been performed to determine hormonal receptors on Bouin's liquid fixed and paraffin embedded tissue. This technique has been evaluated by comparison with conventional immunohistochemistry on frozen sections for 100 surgical biopsy specimens of breast carcinoma Erica (C) Prica (C); in 77 of these cases it was compared with enzyme immunoassay analyses (ER-EIA, PR-EIA). After a pretreatment by proteinase K for ER, the two specific monoclonal antibodies ER or PR were incubated overnight and revealed with a streptavidin biotin complex. The specific staining for ER and PR were exclusively located in the nuclei of cancer cells in both paraffin for ER and PR were exclusively located in the nuclei of cancer cells in both paraffin and frozen sections. The results on paraffin-embedded material were compared successively with ERICA (C) and ER-EIA and with PRICA (C) and PR-EIA. In 100 breast infiltrating carcinomas studied simultaneously on both paraffin and frozen sections, the concordance was 82% for ER and 85% for PR. In 77 breast carcinoma studied simultaneously on paraffin sections and EIA, the concordance was 74% for ER and 79.2% for PR. These results suggest that in the absence of frozen material, an immunohistochemical method on paraffin-embedded material can be used with a lower sensibility than on frozen sections. However, the detection of both ER and PR allows the identification of the hormondependent cases.

Adiponectin values are unchanged during pregnancy in rats.

Adiponectin is believed to be a key factor in determining insulin sensitivity. In turn, insulin sensitivity is known to change from an enhanced state in early pregnancy to a reduced one in late pregnancy. A role for adiponectin in these changes has been proposed for mice but questioned for humans. We addressed this issue in rats by measuring adiponectin expression in both visceral and subcutaneous white adipose tissue, together with tissue content and release of the hormone in non-pregnant and in pregnant rats by days 8, 15 and 19 of pregnancy. Plasma concentration was also determined. No differences were found in any of the parameters measured between non-pregnant and pregnant rats at any time of pregnancy despite changes in white adipose tissue mass and insulin sensitivity. Adiponectin was also detected in cerebrospinal fluid at a concentration 1,000 times lower than in plasma, but again no differences were found between non-pregnant and pregnant animals. It is concluded that adiponectin does not play any role in regulating changes in insulin sensitivity during pregnancy in rats.

[Amniotic band syndrome: description of 2 clinical cases and discussion on etiopathogenesis]. / Síndrome de bridas amnióticas: descripción de dos casos clínicos y discusión etiopatogénica.

The paper reports two cases in which infants were born with alterations described as the Amniotic Band Syndrome. The first one shows inferior limbs in bizarre positions, and the superior mutilated; also a cleft lip was present. The second case corresponds to a newborn with a facionucal band that almost separates the mouth from the rest of the face. Pathogenic theories are reexamined in face of the cases presented.

Influence of continuous electromagnetic fields on the stage, weight and stature of the chick embryo.

The influence of continuous electromagnetic fields (0, 181 or 361 Gs/cm2) on the development of chick embryo (n = 144) was studied. Several parameters were determined at days 5, 10 and 15 of incubation: stage (following Hamburger and Hamilton), vertex-coccyx length (size) and weight. At 5 days of incubation, all embryos showed a similar stage. However, at days 10 and 15, the embryos exposed to 181 Gs/cm2 showed a stage significantly superior to that of the others. There were no differences between the exposed embryos and the control ones with regard to weight and stature, except at 15 days when the embryos exposed to 361 Gs/cm2 showed greater weight and stature than those of the controls.

Effects of static electromagnetic fields on chick embryo pineal gland development.

The effects of static electromagnetic fields on the development of the chick embryo pineal gland were studied. A total of 144 fertilized White Leghorn eggs were sacrificed after 5, 10 and 15 days of incubation. The stage of development was determined in all embryos using the Hamburger and Hamilton method [J Morphol 49: 88-92, 1951]. The various morphometric parameters (diameter and distance of the pineal gland and its lumen) were measured on serial 7-micron-thick sections. The data were obtained in a morphometer and processed statistically. The intensities of the static electromagnetic fields were 18 and 36 mT. Control and exposed embryos were equally distributed and randomly assigned. After 5 days of incubation, 25% of embryos exposed to a static electromagnetic field of 18 mT had a more advanced stage of development than controls and embryos exposed to 36 mT. On the 10th and 15th day, embryos exposed to either 18 or 36 mT tended to be more developed than controls. In the morphometric study, results were similar for the controls and exposed embryos after 5 and 10 days of incubation. However, the values of the 15-day-old embryos exposed to static magnetic fields were lower than the values of the controls (p > 0.01). These differences were more pronounced in the embryos exposed to 36 mT. These results seem to indicate that static electromagnetic fields affect the development and growth of embryos unequally, and that their action can depend not only on the intensity of the static electromagnetic field, but also on the length of exposure and the organ which is developing. It may be interesting to use these data in ultrastructural and physiological studies.

Distribution of estrogen and progesterone receptors in healthy tissue adjacent to breast lesions at various stages--immunohistochemical study of 107 cases.

The purpose of this study was to determine the distribution of ER+ (estrogen receptor) and PR+ (progesterone receptor) epithelial cells in normal mammary tissue or in tissue in contact with or involved in benign or malignant processes. Three important findings emerged from this study. First, a true dissociation was observed between ER+ and PR+ cells in mammary tissue. In premenopausal women some cells express only progesterone receptors. In premenopausal normal tissue, regardless of the menstrual cycle status, 6% of cells are ER+ and 29% PR+. Second, during the menstrual cycle the percentage of positive cells varies. This finding would indicate a change in cell recruitment rather than in intracellular levels. Finally, specific changes in the proportion of positive cells in normal tissue in contact with epithelial proliferations were noted. This finding suggests the possibility of either a diffusible factor or a cellular pathological process spreading beyond areas displaying morphological changes.

Pattern of polyomavirus replication from infection until tumor formation in the organs of athymic nu/nu mice.

Inbred athymic nu/nu BALB/c mice were injected subcutaneously with the highly oncogenic polyomavirus A2 strain, and the sites of viral DNA replication were determined by whole mouse section hybridization (T. W. Dubensky, E. A. Murphy, and L. P. Villareal, J. Virol. 50:779-783, 1984) and Southern blot analysis. We show that infection is persistent in some epithelial tissues (skin, mammary, and salivary glands), in lymphoid organs (spleen and nodes), and in mesenchymal bone tissue. Only mammary glands and bones were targets for tumor formation. Although the same pattern of infection was observed in males and females, mammary adenocarcinomas were induced exclusively in females, while the frequency of osteosarcomas was similar in both sexes. No viral DNA or lytic lesion was detected in kidney, liver, or lung tissue. The restricted targeting of polyomavirus oncogenicity in nude mice, compared with newborn immunocompetent animals, inoculated via the same route with the same virus strain, therefore does not reflect selective tissue targeting of virus replication. These results further document the influence of the age, immunological status, and genetic background of the host on the pattern of viral infection and tumor formation.

Prognostic value of p53 protein expression in squamous cell carcinoma of the esophagus.

In this study, we investigated endoscopic biopsy material consisting of 76 esophageal squamous cell carcinomas after immunostaining with three p53 monoclonal antibodies (PAb 1801, PAb 240, and DO-7). p53 protein was detected in 51.3% of the tumours. The protein was not expressed in the normal mucosa. Statistical analysis of follow-up data on 54 patients showed that high p53 levels were to a significant degree frequently associated with unfavorable treatment response (p < 0.053) and death (p < 0.05); whereas expression of the protein was not correlated with tumor differentiation, DNA ploidy, S phase value, EGF-R expression, or Ki67 index. The results of these studies suggest that in cases of esophageal carcinoma, immunohistochemical detection of p53 protein can be used to predict outcome.

p53 immunohistochemical analysis in breast cancer with four monoclonal antibodies: comparison of staining and PCR-SSCP results.

The expression of p53 protein was examined in a series of 136 primary breast carcinomas, 106 of which were analysed with a panel of four monoclonal antibodies (MAbs 1801, 240, DO7 and DO1). p53 expression was detected with at least one antibody in 40 tumours (38%), whereas only 15 tumours (14%) were positive with all four antibodies. Some variability in the immunostaining could be observed depending on the antibody used. This was noticeable both for the number of positive cells within a section and for the intensity of staining. We therefore selected a panel of 17 tumour sections (nine were highly positive, three with medium to low staining and five with low to negative staining), which we analysed by polymerase chain reaction-single-strand conformation polymorphism analysis (PCR-SSCP) for the presence of a p53 mutation at the molecular level. Mutations were identified in 15 cases. Therefore the proportion of p53-stained cells does not seem to be an exact representation of the number of cancer cells bearing a mutation within a tumour. A statistically significant correlation was observed between p53 expression, regardless of the number of positive antibodies, and grade III disease (P < 0.0001), oestrogen (P < 0.0001) or progesterone receptor negativity (P = 0.0061), increased Ki 67 index (P = 0.0018), epidermal growth factor receptor (EGFR) positivity (P = 0.0076) and aneuploidy (P = 0.037). No correlation was observed with tumour size or lymph node involvement. In univariate analysis p53 expression was not correlated with disease-free survival, in contrast to the classical prognostic parameters, which were statistically correlated. In this series p53 expression was not a marker of early recurrence.

Influencia de la etnia y el sexo en la ingesta de alimentos como factores de riesgo de malnutrición en escolares / Influence of ethnicity and gender in school children food intake as malnutrition risk factors

La dieta de los niños en nuestro país suele ser deficitaria en frutas y verduras y rica en alimentos hipercalóricos. El objetivo del presente estudio es valorar la frecuencia de la ingesta de los grupos de alimentos en escolares de nuestro medio y la influencia sobre ella del sexo y la etnicidad. Se valoraron en una población de 383 niños de 2.º, 3.º y 4.º de Educación Primaria las características socioeconómicas y demográficas, los estilos de vida y la frecuencia de ingesta de grupos de alimentos. Los niños ingirieron más raciones diarias de alimentos ricos en hidratos de carbono (4,38 ± 1,6 frente a 3,97 ± 1,4; p=0,03) y más lácteos (3,23 ± 1,1 frente a 2,83 ± 1,0; p= 0,002) que las niñas; por otro lado, la niñas consumieron más golosinas que los niños (2,81 ± 3,9 frente a 2,01 ± 2,3 raciones por semana; p= 0,03). Un alto porcentaje de niños no cumplía con las recomendaciones habituales de ingesta por defecto de frutas y verduras (78,1%), lácteos (43,4%) e hidratos de carbono (42,8%), y por exceso de proteicos (8,1%), bollería (21,2%),golosinas (18,9%) y refrescos-snacks (8,7%). Los niños pertenecientes al grupo con etnicidad ingirieron significativamente menos lácteos, alimentos proteicos y fruta-verdura y, sin embargo, más bollería, golosinas y refrescos-snacks que los niños sin etnicidad. En el análisis de la relación entre los distintos grupos de alimentos se han encontrado dos asociaciones significativas (p <0,01): 1) la ingesta de frutas y verduras se asoció a un mayor consumo de hidratos de carbono y de pescado, y aun menor consumo de refrescos-snacks y tiempo frente al televisor; 2) la ingesta de bollería se asoció a un mayor consumo de refrescos/snacks y de golosinas. El estudio concluye que el sexo y la etnicidad son factores que influyen en la dieta de los niños en edad escolar y deben considerarse como factores de riesgo de malnutrición (AU)

Children’s diet in our country seems to be deficient in fruits and vegetables and rich in hyper caloric food. The purpose of this study is to access intake frequency of food groups in school children of our environment and the influence gender and ethnicity pose over it as well. We have evaluated the socioeconomic and demographic characteristics, lifestyles and the intake frequency of food groups in a population of 383 2nd, 3rd and 4th grade children from elementary school. Boys consume more daily servings of food rich in carbohydrates (4.38 ± 1.6 vs. 3.97 ± 1.4; p=0.03) and more dairy products (3.23 ± 1.1 vs. 2.83 ± 1.0; p= 0.002) than girls, and, on the other hand, girls eat more candy than boys (2.81 ± 3.9 vs. 2.01 ± 2.3 servings per week; p= 0.03). A high percentage of children did not meet regular recommended food percentages, both by deficiency: fruits and vegetables (78.1%), dairy products (43.4%) and carbohydrates (42.8%); and by excess: protein food (8.1%), pastries (21.2%), candy (18.9%), and sodas/snacks (8.7%). Children belonging to a certain ethnic group consume significantly less dairy products, protein products and fruits/vegetables; and, at the same time, more pastries, candy and sodas/snacks than children that do not belong to a specific ethnic group. Two significant associations (p <0.01) have been found in the analysis of the relationship between different food groups: 1) fruits and vegetables intake was associated with a higher consumption of carbohydrates and fish, and with alower consumption of sodas/snacks and time watching TV; and2) pastry intake was associated with a higher consumption of sodas/snacks and sweets. The study concludes that gender and ethnicity are factors that influence the school children´s diet and they shall be considered as risk factors for malnutrition (AU)

Caracterización citogenética en cáncer laríngeo / Cytogenetic characterization in laryngeal cancer

El cáncer laríngeo representa un 23 por ciento de las neoplasias malignas en la práctica clínica del otorrinolaringólogo. Mediante técnicas de citogenética se ha logrado describior cambios en el DNA de tumores epiteliales de cabeza y cuello, siendo lo mas frecuentes pérdida de 3p, 8p y 18q. Entre Mayo de 1996 y Julio de 1997 se estudiaron 13 pacientes con diagnóstico de cáncer laríngeo. Las alteraciones encontrada corresponden a pérdida del cromosoma Y, alteraciones del cromosoma 5, 4q+, 2p-y manosomía del cromosoma 15