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Biomedical subjects

A Cano

Publications and source records attributed to A Cano.

At least 91 records · Page 5Linked to original sources

Cadherin/catenin complexes in murine epidermal keratinocytes: E-cadherin complexes containing either beta-catenin or plakoglobin contribute to stable cell-cell contacts.

The cadherin/catenin complexes expressed by a murine epidermal keratinocyte cell line PDV, expressing E- and P-cadherin, have been analysed using a combination of biochemical and confocal microscopy analysis. Two types of E-cadherin complexes, containing beta-catenin or plakoglobin and alpha-catenin, were detected in PDV cells as in other cell types, while P-cadherin was mainly detected in complexes containing beta-catenin and alpha-catenin in PDV and other murine epidermal keratinocytes. Biotin-labelling studies have shown that both types of E-cadherin complexes are present at the surface of confluent cells. Furthermore, confocal microscopy analysis indicated that E-cadherin/plakoglobin complexes are located in stable cell-cell contacts at the middle lateral membranes and associated with alpha-catenin and the actin cytoskeleton, with a similar distribution to that to the E-cadherin/beta-catenin complexes. In addition, E-cadherin/plakoglobin complexes not associated with alpha-catenin or the actin cytoskeleton were detected in lower planes of the lateral contacting membranes as well as E-cadherin non-associated with catenins in the more basal planes. These studies support that in murine epidermal keratinocytes both beta-catenin- and plakoglobin-containing E-cadherin complexes contribute to the maintenance of stable cell-cell contacts and suggest a differential role of the plakoglobin containing complexes in different epithelial cell types.

Animals↗

Down-regulation of E-cadherin in mouse skin carcinoma cells enhances a migratory and invasive phenotype linked to matrix metalloproteinase-9 gelatinase expression.

To assess the role of gelatinases in mouse skin tumor progression and their link to the expression of E-cadherin (E-CD), the cell-cell adhesion protein, we used the highly metastatic squamous HaCa4 cell line and several HaCa4-derived clones obtained by transfection of the mouse E-CD cDNA. Expression of matrix metalloproteinase-9 (MMP-9) mRNA and protein activity were present in E-CD (-) HaCa4 and control clones in culture, but they were strongly diminished in E-CD (+) clones (E24 and E62) at subconfluence. To explore the suppressive effect of the cell-cell contacts mediated by E-CD on MMP-9 expression, we introduced a plasmid encoding mouse E-CD antisense cDNA into the E24 cell clone. The transfectant P1-clones obtained with reduced or absent E-CD expression showed increased levels of MMP-9 gelatinase, motility in vitro, and metastatic potential in vivo. Expression of MMP-9 in the various cell clones was also negatively modulated by cell density, but this effect was much stronger in E-CD (+) cells, despite the fact that all of the cell clones analyzed maintained the expression of P-cadherin and made cell-cell contacts at high cell density. Our results indicate that in this cell system, the E-CD-mediated cell-cell contacts are involved in the down-regulation of MMP-9 expression. Thus, the loss of E-CD triggers a migratory and invasive phenotype in mouse squamous carcinoma cells.

Animals↗

Idiopathic biliary ductopenia in adults without symptoms of liver disease.

BACKGROUND: Idiopathic adulthood ductopenia is a severe cholestatic liver disease of unknown cause characterized by loss of the interlobular bile ducts in more than 50 percent of the portal tracts. In most reported cases, cirrhosis and liver failure develop. METHODS: We studied 24 adults with abnormal results on liver-function tests but no symptoms of liver disease. All had liver biopsies that showed a lack of bile ducts in many of the portal tracts. RESULTS: The 17 women and 7 men had a mean age of 41 years (range, 27 to 57). All were asymptomatic and had high serum gamma-glutamyltransferase concentrations (mean [+/-SD], 179 +/- 84 U per liter); 75 percent also had abnormal serum alanine aminotransferase concentrations. The proportion of portal tracts that had bile ducts was 62 +/- 7 percent (range, 55 to 78 percent). Three patients had a second liver biopsy three to nine years after the first; there were no changes over time. In four of the five patients treated with 600 to 900 mg of ursodiol two to three times daily, results of liver-function tests returned to normal. CONCLUSIONS: Idiopathic biliary ductopenia, with an apparently nonprogressive clinical course, can occur in adults who have no symptoms of biliary disease.

Adult↗

Analysis of the E-cadherin and P-cadherin promoters in murine keratinocyte cell lines from different stages of mouse skin carcinogenesis.

We previously isolated the 5' upstream sequences of the mouse P-cadherin gene, in which putative binding sites for several transcription factors were identified between nt-101 and +30. In the study reported here, the promoter activity of the postulated 5' cis-acting sequences of the P-cadherin promoter, and the activity of the proximal E-cadherin promoter were investigated in several murine keratinocyte cell lines showing different levels of P- and E-cadherin expression as well as different morphology and tumorigenic behavior. Cell-type specificity and optimal activity of P-cadherin expression in murine keratinocytes was conferred by 5' sequences located between nt -200 and +30, and the GC-rich region (nt -101 to +80) and a CCAAT box element (nt -65) had a major regulatory role. The cell-type specificity of the E-cadherin promoter, on the other hand, was mediated by a combination of positive regulatory elements, a GC-rich region (nt -58 to -24), and a CCAAT box (nt -65) and repressor elements inside the E-pal sequence. Interestingly, the maximum repressor effect of the E-pal element was observed in non-expressing undifferentiated spindle cells. In vitro binding studies indicated that the GC-rich region of the P-cadherin promoter was mainly recognized by Sp1-related nuclear factors, whereas both AP2- and Sp1-related factors were involved in the interaction of the GC-rich region of the E-cadherin promoter. Common factors (probably related to the CP1 family) seemed also to be involved in the recognition of the CCAAT-box element of both the E- and P-cadherin promoters, but additional specific factors participated in the interaction with the CCAAT box of the E-cadherin promoter. Our studies also support the hypothesis that loss or modification of some of the regulatory factors occurs during mouse skin tumor progression.

Animals↗

Efficacy of a dating violence prevention program on attitudes justifying aggression.

OBJECTIVES: The purpose of this pilot study was to evaluate a five-session dating violence prevention curriculum in terms of its effect on attitudes justifying the use of dating violence. METHODS: The curriculum was implemented in all health classes in a Long Island, New York, school. A total of 193 students participated (boys, n = 106; girls, n = 87). A quasi-experimental design was used to evaluate change in attitudes justifying dating violence, with health classes randomly assigned to the treatment or no-treatment conditions. RESULTS: Pre- to postprogram assessments indicated that there were significant decreases in overall attitudes justifying the use of dating violence as a means to resolve conflict among students exposed to the curriculum material, whereas those who were not exposed did not show attitude change from pre- to postprogram evaluation. CONCLUSIONS: The curriculum shows promise as an effective tool for changing attitudes condoning dating violence. Future research is needed to determine whether the observed attitude change is also linked to reduction in aggressive behaviors.

Adolescent↗

Romantic jealousy and affairs: research and implications for couple therapy.

Romantic jealousy is often experienced after a situation is interpreted as threatening to one's intimate relationship and can involve elements of other emotions such as anger, fear, and sadness. An individual experiencing jealousy may engage in a number of behaviors and thought processes aimed at reducing jealousy or keeping the relationship intact. However, these cognitions and behaviors may not achieve either of these goals and may escalate problems in the relationship. We review the theoretical and empirical jealousy literature and integrate the research findings with existing models of couple therapy. Assessment and treatment guidelines are offered on the basis of theory, research, and clinical practice. Future research directions are also discussed.

Adaptation, Psychological↗

Lung function and airway responsiveness in children and adolescents after hyaline membrane disease: a matched cohort study.

Some investigators consider prematurity to be responsible for the lung function abnormalities found in prematurely born children and adolescents who had neonatal respiratory diseases. This study attempts to measure the effect of neonatal respiratory disease on lung function during school age and adolescence, by controlling the confounding effect due to prematurity. Lung volumes, airway resistance and specific airway conductance measured by plethysmography, maximum expiratory flow-volume curves, pulmonary diffusion of carbon monoxide, and the airway responsiveness to a challenge with methacholine, were determined in a cohort of children aged 8-14 yrs, who had suffered from hyaline membrane disease but who did not develop bronchopulmonary dysplasia. The values obtained were compared with those of children without hyaline membrane disease, not ventilated for other causes, and matched for gestational age, sex and age. Thirty six pairs of children were enrolled, of which 26 participated in the methacholine test. Compared to their paired controls, children with hyaline membrane disease had a significantly lower forced expiratory volume in one second (FEV1), forced mid-expiratory flow (FEF25-75), and maximal expiratory flow when 75, 50 and 25% of the forced vital capacity remained in the lung (MEF75, MEF50 and MEF25, respectively), and a significantly higher airway resistance (Raw). The effect was less in children born more prematurely, who showed less difference in FEF25-75, MEF75 and MEF25. The duration of treatment with steroids in the neonatal period was associated with a reduction in the differences in FEV1, MEF25 and Raw. Independent of prematurity, hyaline membrane disease and its treatment is associated with alterations in long-term lung function, even in children who do not develop bronchopulmonary dysplasia. The effect can be less in more premature children, and neonatal steroids can have a long-term preventive effect.

Adolescent↗

Up-regulation of vascular endothelial growth factor/vascular permeability factor in mouse skin carcinogenesis correlates with malignant progression state and activated H-ras expression levels.

Angiogenesis is a crucial process for tumor growth and metastasis regulated by the balance of positive and negative factors. Vascular endothelial growth factor (VEGF/VPF) is a specific mitogen for endothelial cells that has been shown to be overexpressed in a variety of tumors and other inflammatory diseases. To analyze the implication of VEGF/VPF during tumorigenesis, we have studied its expression at different stages of tumor development using the mouse skin carcinogenesis model. VEGF/VPF mRNA was induced in skin in vivo after 12-O-tetradecanoylphorbol-13-acetate treatment. Constitutive up-regulation of VEGF/VPF at the mRNA and protein levels was also observed in premalignant papillomas and, at a higher level, in squamous carcinomas, suggesting a correlation between VEGF/VPF expression and tumor progression. A direct positive correlation between VEGF/VPF mRNA expression and the level of activated H-ras gene was found in a series of cell lines representing different stages of epidermal tumor development. Consequently, a clone of one of these cell lines, HaCa4, which has lost most of its v-ras expression, down-regulated VEGF mRNA expression concomitantly with its metastatic potential. Direct evidence of H-ras involvement in VEGF induction was obtained when an immortalized mouse keratinocyte cell line transduced with a retrovirus carrying v-H-ras showed highly increased VEGF/VPF mRNA levels. These data show that in mouse skin carcinogenesis, the VEGF/VPF angiogenic stimulus occurs early during premalignant papilloma development and further increases at later stages. Moreover, we demonstrate that increasing the activated H-ras dose, a phenomenon that takes place sequentially throughout mouse skin tumor development, may play an additional role by facilitating malignant in vivo progression through the modulation of VEGF/VPF-mediated angiogenesis.

Animals↗

Carcinoma cell lines become sensitive to DNA-damaging agents by the expression of the adenovirus E1A gene.

Squamous cell carcinomas can show different oncogenic alterations, histological patterns, and an unpredictable clinical behavior. We previously reported that the adenovirus E1a gene may induce sensitivity to DNA-damaging agents in mouse keratinocytes. In order to study whether E1a expression could be used as a therapeutic agent in different malignant cell lines carrying mutations on the p53 gene and other oncogenic alterations, we transfected and infected several murine and human carcinoma cell lines (HaCa4; MSC11A5; HeLa) with vectors containing the 13S or 12S E1a region. We evaluated the sensitivity to cisplatin (CDDP), doxorubicin (DOX) and gamma irradiation (RX) by the crystal violet method. The induction of apoptosis was assessed by flow cytometry and presence of DNA ladders in agarose gels. The expression of E1a and the tumor suppressor p53 protein was analysed by Western blotting. The carcinoma cell lines expressing E1a were about four- to tenfold more sensitive to CDDP and RX, respectively, than the control cells. Moreover, the reduction in cell viability and cell growth after exposure to CDDP or RX was very significant in the carcinoma cells expressing E1a. With these results, we conclude that expression of E1a may confer great sensitivity to DNA-damaging agents on squamous cell carcinoma cells independently of the p53 protein status and other oncogenic alterations.

Adenoviridae↗

Inhibition by L-ascorbic acid and other antioxidants of the 2.2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) oxidation catalyzed by peroxidase: a new approach for determining total antioxidant status of foods.

The accumulation of 2.2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical catalyzed by peroxidase can be inhibited by the presence of L-ascorbic acid in the reaction medium, this inhibition delaying the accumulation of the ABTS radical and giving rise to a lag time. A kinetic approach to explain this lag time is presented, which also makes it possible to determine the amount of L-ascorbic acid in the reaction medium. The stoichiometry of the system was determined as 1 mol of L-ascorbic reducing 2 mol of ABTS radicals. L-Ascorbic acid is not the only compound to have this ability, since other antioxidant compounds also react with the ABTS radical. We studied the ABTS/H2O2/horseradish peroxidase system in the presence of L-ascorbic acid and other antioxidant compounds. The influence of such factors as pH, enzyme concentration, and L-ascorbic acid concentration was studied. A good correlation between the lag time and the L-ascorbic acid present in the medium was observed, and under optimal conditions, the method could determine as little as 0.65 nmol of L-ascorbic acid. Based on our findings, we propose a method to measure the total antioxidant activity of different compounds related to L-ascorbic acid and apply this method to determining the total antioxidant activity present in fruit juices.

Antioxidants↗

Endothelin(B) receptor activates NHE-3 by a Ca2+-dependent pathway in OKP cells.

To examine the mechanisms by which endothelin (ET) regulates the Na/H antiporter isoform, NHE-3, OKP cells were stably transfected with ET(A) and ET(B) receptor cDNA. In cells overexpressing ET(B), but not ET(A) receptors, ET-1 increased Na/H antiporter activity (JNa/H). This effect was inhibited by a nonselective endothelin receptor blocker and by a selective ET(B) receptor blocker but was not inhibited by an ET(A) selective receptor blocker. In ET(B)-overexpressing cells, 10(-8) M ET-1 inhibited adenylyl cyclase, but protein kinase A inhibition and pertussis toxin pretreatment did not affect Na/H antiporter activation by ET-1. ET-1 caused a transient increase in cell [Ca2+], followed by a sustained increase. Increases in cell [Ca2+] were partially inhibited by pertussis toxin. ET-1-induced increases in J(Na/H) were 50% inhibited by clamping cell [Ca2+] low with BAPTA, and by KN62, a Ca-calmodulin kinase inhibitor. Inhibitors of protein kinase C, cyclooxygenase, lipoxygenase, and cytochrome P450 and cyclic GMP were without effect. In ET(A)-overexpressing cells, ET-1 increased cell [Ca2+] but did not increase JNa/H. In summary, binding of ET-1 to ET(B) receptors increases Na/H antiporter activity in OKP cells, an effect mediated in part by increases in cell [Ca2+] and Ca-calmodulin kinase. Increases in cell [Ca2+] are not sufficient for Na/H antiporter activation.

Animals↗

[Ambulatory care of chronic headache secondary to abuse of ergot preparations].

OBJECTIVE: To analyse the results of out-patient treatment with diminishing doses of oral dihydroergotamine for patients with chronic migraine resulting from ergotics abuse. DESIGN: A prospective and descriptive intervention study. SETTING: County hospital. Neurology out-patient clinic. PATIENTS: All the patients with chronic migraine as a side-effect of abuse of ergotic preparations who were referred to neurology out-patients over 18 months. INTERVENTIONS: 1) Patients were told verbally of the causes of their migraine; 2) suppression of the ergotics, using diminishing doses of dihydroergotamine (Dihydergot gotas); 3) Prophylactic treatment with flunarizine (Sibelium); 4) Symptomatic treatment with sodic naproxen (Antalgin 550); 5) monthly and three-monthly check-up. MEASUREMENTS AND MAIN RESULTS: Over 18 months 25 patients with migraines due to ergotic abuse (6.7% of total migraines) were included. 4 were men; 21 women. Their average age was 43 (SD: 13). At one month the response was excellent in 19 patients (76%), good in 3 (12%) and bad in 3 (12%). At three months, the response was excellent in 17 patients (68%), good in 4 (16%), bad in 2 (8%) and 2 did not attend for the check-up (8%). CONCLUSIONS: 1) Out-patient treatment with diminishing doses of oral dihydroergotamine is effective in treating chronic migraine due to ergotics abuse. 2) The general practitioner should intervene in the identification and prevention of ergotics abuse.

Administration, Oral↗

Expression pattern of the cell adhesion molecules. E-cadherin, P-cadherin and alpha 6 beta 4 intergrin is altered in pre-malignant skin tumors of p53-deficient mice.

Expression of the cell adhesion molecules E-cadherin, P-cadherin and alpha 6 beta 4 integrin and of the keratin K13 has been analyzed in chemically induced benign skin papillomas with genetically pre-determined risks for malignant conversion. It has been previously shown that papillomas induced in mice lacking both alleles of the p53 gene have a much higher rate of malignant conversion than those induced in wild-type and heterozygous p53 mice. Alterations in the expression pattern of the E-cadherin molecule, including focal loss at cell-cell contacts and heterogeneous distribution in the differentiated layers, were found in about 70% of the p53 null papillomas. In contrast, all of the wild-type and over 85% of the heterozygous p53 papillomas exhibited an expression pattern of E-cadherin indistinguishable from that of normal epidermis. Alterations in P-cadherin expression were also detected in the p53 null papillomas: aberrant suprabasal localization and heterogeneous distribution were observed more frequently than in heterozygous and wild-type p53 papillomas. The alpha 6 beta 4 integrin showed suprabasal expression in more than 70% of the papillomas derived from either wild-type, heterozygous or homozygous p53 null mice. Surprisingly, the extent of the suprabasal localization of alpha 6 beta 4 decreased in the p53 null papillomas. Aberrant keratin K13 expression was also detected in the majority of cases of all p53 genotypes, but again there was a clear decrease in expression levels in the p53 null papillomas. These alterations were also associated with keratinocytic atypia, which increased significantly in the p53 null papillomas. Changes in these parameters were particularly evident during malignant conversion in invasive regions of one progressing p53 null papilloma. Our results indicate the existence of dynamic changes in the expression pattern of the 3 cell adhesion molecules analyzed and identify down-regulation of E-cadherin as an early step in malignant conversion.

Animals↗

Suppression of the metastatic phenotype of a mouse skin carcinoma cell line independent of E-cadherin expression and correlated with reduced Ha-ras oncogene products.

The HaCa4 cell line, derived from a mouse skin carcinoma induced by Harvey murine sarcoma virus, is highly tumorigenic when injected into nude mice and produces multiple metastases in the lungs. HaCa4 cells express high levels of viral Ha-ras oncogene products, anomalously synthesize the embryonic/simple epithelial keratin K8, and have lost the expression of the cell-cell adhesion receptor E-cadherin (E-CD). E-CD(+) cell clones (E62 and E24), obtained by transfection of an exogenous E-CD cDNA into HaCa4 cells, had a decreased ability to migrate through type IV collagen matrices. However, the E-CD (+) E62 clone remained as metastatic as the parental cell line, whereas the E24 clone, which does not take up the exogenous cDNA but spontaneously switches on the endogenous E-CD gene, suppressed the metastatic phenotype although it maintained its tumorigenicity. E24 cells had fivefold to sixfold lower levels of viral Ha-ras mRNA and p21 protein than the other cell lines. In addition, they did not synthesize K8 but rather switched on keratin K19. The comparison of E-CD proteins synthesized by E62 and E24 cell lines revealed no structural or functional differences because both localized at cell-cell contacts and associated with alpha-catenin, beta-catenin, and plakoglobin. Furthermore, E-CD was still expressed in metastatic lung nodules produced by E62 cells. These results suggest that suppression of the metastatic phenotype in E24 cells occurs independently of E-CD expression and correlates with decreased levels of the oncogenic ras p21 protein.

Animals↗

Acoustic trauma caused by lightning.

Lesions produced by exposure to noise are frequent in everyday life. Injuries may be found in all systems of the human body, from the digestive to the endocrine, from the cardiovascular to the nervous system. Many organs may be damaged, the ear being one of them. It is known that noise produced by factories, airports, musical instruments and even toys can cause auditory loss. Noises in nature can also cause acoustic trauma. This report is the case history of acoustic trauma caused by lightning. The patient was studied with CAT scan, electroencephalogram, and brain mapping, impedance audiometry with tympanogram and acoustic reflex, audiometry and evoked otoacoustics emissions: distortion products and transients.

Acoustic Impedance Tests↗