[Effect of combination therapy with hypothermia, magnesium and tirilazad in an experimental model of diffuse cerebral ischemia]. / Efecto de la hipotermia combinada con magnesio y tirilazad en un modelo experimental de isquemia cerebral difusa*

OBJECTIVE: To determine the grade of neuroprotection of combined treatment with moderate hypothermia, tirilazad and magnesium sulfate. Cerebral ischemia is one of the problems of great interest at present, with limited therapeutic measures. Hypothermia, one of the more efficient measures, together with neuroprotector pharmaceuticals, could be a valid alternative. DESIGN: Experimental study with a control group and two levels of application of therapeutic measures. CONTEXT: Experimental laboratory of the Medicine Faculty. PARTICIPANTS AND METHOD: Twenty-eight Wistar rats underwent global cerebral ischemia of 10 minutes duration by the combination of bilateral carotid clamping and controlled hypotension (mean arterial pressure: 45 mmHg). Three groups were used: group I, normothermia maintenance; group II, moderate hypothermia (32-33 degrees C) for 2 hours; group III, hypothermia and administration of tirilazad mesylate and magnesium sulfate during the reperfusion and two hours after ischemia. The animals were sacrificed at 7 days and, after processing the tissue, the neurons preserved in layer CA1 of the hippocampus were counted. RESULTS: There is a significantly greater neuronal preservation in group III with regard to group I (55.4 +/- 5.1 versus 38.7 +/- 8.8, p < 0.0001). If we compare groups II and III, significant differences are only obtained on the right side and in the hippocampus considered globally, favoring the group with hypothermia and drugs. When groups I and II are compared there are no significant differences. CONCLUSIONS: Association of moderate hypothermia, magnesium sulfate and tirilazad mesylate in the experimental model of transitory global ischemia used is confirmed as an effective neuroprotector measure, surpassing the degree of neuronal preservation of hypothermia alone.

Expresión de E-cadherina, laminina y colágeno IV en la evolución de displasia a carcinoma epidermoide oral / E-cadherin, laminin and collagen IV expression in the evolution from dysplasia to oral squamous cell carcinoma

Objetivos: Estudiar la pérdida o reducción de la adhesión celular mediada por E-cadherina en leucoplasias, carcinomas epidermoides y metástasis ganglionares. Estudiar la pérdida de continuidad de la expresión de laminina y colágeno IV en la membrana basal epitelial en el desarrollo biológico de las leucoplasias y carcinomas orales.Material y metodo: Hemos estudiado 124 muestras de pacientes portadores de leucoplasias y carcinomas orales con diversos diagnósticos que abarcan desde epitelio normal (13 muestras), displasias leves (2), displasias moderadas (12), carcinomas in situ (13) carcinomas microinvasores (11) Carcinoma epidermoide oral (64 muestras) y metástasis ganglionar(9). Se construyeron 7 bloques de tissue microarrays con aguja de 2mm y se realizó un estudio mediante técnica inmunohistoquímica para E-cadherina (clona 36, T.D. ABD Company), Laminina (078P, Biogenex) y Colágeno IV (PHM12, Biogenex).Resultados: En Displasias Leves y Moderadas presentan pérdida de expresión de E-cadherina, Laminina, y ColágenoIV (20%). En Carcinomas in situ y Microinvasores, presentaron pérdida de expresión de E-cadherina (73%), y en Laminina y Colágeno IV (57%). En los carcinomas epidermoides, encontramos pérdida de expresión de E-cadherina (90%) y discontinuidad en la M. basal (70%). Todas las metástasis ganglionares presentaron pérdida de E-cadherina y discontinuidad en Laminina y Colágeno IV.Conclusiones: La pérdida de expresión de E-cadherina se incrementa al aumentar el grado de displasia de las lesiones. La perdida de continuidad en la expresión de laminina y Colágeno IV sigue una evolución paralela desde displasias a metástasis ganglionares. La disminución en la expresión de los tres marcadores ha sido significativa en la evolución de las lesiones orales

Objectives: Study the loss or reduction of the cellular adhesion mediated for E-cadherin in oral leukoplakias, oral squamouscell carcinomas and metastatic nodules. Study the loss of continuity of the laminin and collagen IV expression in the epithelial basal membrane from the biological development of the oral leukoplakias and oral carcinomas.Material and method: we have studied 124 samples of patient payees leukoplakias and oral carcinomas with diverse diagnosis that embrace from normal epithelium (13 samples), mild dysplasias (2), moderate dysplasias (12), “in situ” carcinomas (13), microinvasive carcinomas (11) oral squamous cell carcinomas (64 samples) and metastatic nodules (9). 7 blocks of tissue microarrays were built with needle of 2mm and was carried out a study by means of immunohistochemicaltechnique for E-cadherin (clone 36, Biogenex), Laminin (078P, Biogenex) and Collagen IV (PHM12, Biogenex).Results: In Mild and Moderate Dysplasias the results present loss of E-cadherin, Laminin, and Collagen IV (20%) expression. “in situ” and microinvasive carcinomas, the results presented loss of E-cadherin expression (73%), and loss in Laminin and Collagen IV expression (57%). In the squamous cell carcinomas , we find E-cadherin underexpression (90%) and discontinuity in the Basal Membrane. (70%). All the metastatic nodules presented loss of E-cadherin expressionand discontinuity in Laminin and Collagen IV expression.Conclusions: The loss of E-cadherin expression is increased when increasing the dysplasia grade of lesions. The loss of continuity in the laminin and Collagen IV expression follow a parallel evolution from dysplasias to metastatic nodules. The underexpression of the three markers has been significant in the evolution of the oral lesions

E-cadherin, laminin and collagen IV expression in the evolution from dysplasia to oral squamous cell carcinoma.

OBJECTIVES: Study the loss or reduction of the cellular adhesion mediated for E-cadherin in oral leukoplakias, oral squamous cell carcinomas and metastatic nodules. Study the loss of continuity of the laminin and collagen IV expression in the epithelial basal membrane from the biological development of the oral leukoplakias and oral carcinomas. MATERIAL AND METHOD: we have studied 124 samples of patient pays leukoplakias and oral carcinomas with diverse diagnosis that embrace from normal epithelium (13 samples), mild dysplasias (2), moderate dysplasias (12), in situ carcinomas (13), microinvasive carcinomas (11) oral squamous cell carcinomas (64 samples) and metastatic nodules (9). 7 blocks of tissue microarrays were built with needle of 2mm and was carried out a study by means of immunohistochemical technique for E-cadherin (clone 36, Biogenex), Laminin (078P, Biogenex) and Collagen IV (PHM12, Biogenex). RESULTS: In Mild and Moderate Dysplasias the results present loss of E-cadherin, Laminin, and Collagen IV (20%) expression. in situ and microinvasive carcinomas, the results presented loss of E-cadherin expression (73%), and loss in Laminin and Collagen IV expression (57%). In the squamous cell carcinomas , we find E-cadherin underexpression (90%) and discontinuity in the Basal Membrane. (70%). All the metastatic nodules presented loss of E-cadherin expression and discontinuity in Laminin and Collagen IV expression. CONCLUSIONS: The loss of E-cadherin expression is increased when increasing the dysplasia grade of lesions. The loss of continuity in the laminin and Collagen IV expression follow a parallel evolution from dysplasias to metastatic nodules. The underexpression of the three markers has been significant in the evolution of the oral lesions.

Expresión proteica de p53 y proliferación celular en leucoplasias orales / Proteic expressión of p53 and cellular proliferation in oral leukoplakias

Objetivos: Conocer la expresión proteica de las alteraciones genéticas que se producen en las etapas precoces de la cancerización del campo de cavidad oral en nuestro medio. Estudiar la proliferación celular mediante Ki-67 y la expresión de la proteína p53 para valorar si las alteraciones en la expresión proteica de estos marcadores suceden de forma secuencial a través de las distintas etapas en la cancerización del campo de la cavidad oral. Material y métodos: Se realizó un estudio mediante técnicas de inmunohistoquímica sobre 53 pacientes que presentaron lesiones de leucoplasia oral, atendidos por el Servicio de O.R.L del Hospital Universitario de Salamanca, desde 1.990 hasta 2000. Se incluyen en el estudio 11 muestras de epitelio normal, 15 displasias leves y moderadas, 15 carcinomas in situ, y 12 carcinomas microinvasores. Resultados: Encontramos la proliferación celular aumentada y sobreexpresión de p53 a medida que avanzamos en el grado de severidad histopatológica de las lesiones. Las alteraciones más precoces son el aumento significativo de la proliferación celular en displasias leves y moderadas y el aumento de expresión de p53. Conclusión: La leucoplasia oral es un estado precanceroso quec onstituye una lesión cancerizable debido a las alteraciones genéticas que intervienen en la evolución de la lesión. El estudio inmunohistoquímico y molecular de las lesiones es un medio rutinario que permite conocer la expresión proteica de las alteraciones genéticas, que puede ayudar en el diagnóstico precoz y tratamientode esta patología, teniendo especial relevancia el estudio de Ki-67 en etapas iniciales y p53 en lesiones más avanzadas

OBJECTIVES: We intend to know the protein expression of genetic alterations that take place in the early stages in the field cancerization of oral cavity in our means as well as to study the cellular proliferation by means of Ki-67 and the protein product expression of p53 to value if the alterations in the protein products expression of these markers happen in a sequential pathway through the different stages in the field cancerization of oral cavity. MATERIALS AND METHODS: A study was made by immunohistochemistry on 53 patients that presented lesions of oral leukoplaquia, assisted by the ENT service at University Hospitalof Salamanca, from 1.990 up to 2000. 11 samples of normal epithelium,15 mild to moderate dysplasias, 15 in situ carcinomas and 12 microinvasive carcinomas are included in the study. RESULTS: we find an increased cellular proliferation and p53 over-expression as we advance in the grade of severity histopathologic of these lesions. The most early alterations are a significant increase of cell proliferation in mild and moderate dysplasias and an increased p53 over-expression. CONCLUSIONS: Oral leukoplaquia is a precancerous stage that constitutes a canzerisable lesion due to the genetic alterations that mediate in the evolution of lesion. Routine Immunohistochemical and molecular study of these lesions allow us to know the protein expression of genetic alterations that can help in the early diagnosis and treatment of this pathology, having special relevance the study of Ki-67 in early stages and p53 inadvanced lesions

Proteic expression of p53 and cellular proliferation in oral leukoplakias.

OBJECTIVES: We intend to know the protein expression of genetic alterations that take place in the early stages in the field cancerization of oral cavity in our means as well as to study the cellular proliferation by means of Ki-67 and the protein product expression of p53 to value if the alterations in the protein products expression of these markers happen in a sequential pathway through the different stages in the field cancerization of oral cavity. MATERIALS AND METHODS: A study was made by immunohistochemistry on 53 patients that presented lesions of oral leukoplaquia, assisted by the ENT service at University Hospital of Salamanca, from 1.990 up to 2000. 11 samples of normal epithelium, 15 mild to moderate dysplasias, 15 in situ carcinomas and 12 microinvasive carcinomas are included in the study. RESULTS: we find an increased cellular proliferation and p53 over-expression as we advance in the grade of severity histopathologic of these lesions. The most early alterations are a significant increase of cell proliferation in mild and moderate dysplasias and an increased p53 over-expression. CONCLUSIONS: Oral leukoplaquia is a precancerous stage that constitutes a cancerisable lesion due to the genetic alterations that mediate in the evolution of lesion. Routine Immunohistochemical and molecular study of these lesions allow us to know the protein expression of genetic alterations that can help in the early diagnosis and treatment of this pathology, having special relevance the study of Ki-67 in early stages and p53 in advanced lesions.

[Atypical scrotal leiomyoma. A case report]. / Leiomioma atípico de escroto. A propósito de un caso.

A case of intrascrotal atypical leiomyoma is reported. This lesion is extremely rare and usually misdiagnosed. Microscopically, contain bizarre and pleomorphic tumor cells, as well as showed immunohistochemical evidence of smooth muscle differentiation. An important microscopic criterion in the distinction with other entity, as the leiomyosarcoma, is the number of mitotic figures, scarce in the atypical leiomyoma and high in the smooth muscle tumours malignant.

Analysis of bcl-2 in sporadic breast carcinoma.

BACKGROUND: The bcl-2 gene encodes a protein that blocks apoptosis and might help to promote tumor development. It is expressed in a high percentage of breast tumors and is associated with good prognostic features. However, the mechanisms that regulate bcl-2 expression in breast carcinoma are unknown. Moreover, immunohistochemical detection of bcl-2 is related inversely to p53 expression. This notwithstanding, the immunohistochemical detection of p53 does not always correlate with the detection of p53 gene mutations. The authors studied the molecular organization of bcl-2 as well as the methylation status of its CpG island and analyzed the correlation between bcl-2 expression and p53 gene mutations. METHODS: The molecular organization of the bcl-2 gene and the methylation pattern of its CpG island were analyzed by Southern blot analysis. In addition, immunohistochemical analysis of bcl-2 and p53 protein expression was performed. Finally, the presence of mutations at exons 5-9 of the p53 gene were analyzed by polymerase chain reaction and single-strand conformation polymorphism. RESULTS: No molecular abnormality was found at the bcl-2 locus in cases of sporadic breast carcinoma. Moreover, loss of heterozygosity analysis failed to detect any allelic loss in the study cases. It also was found that the bcl-2 CpG island was demethylated in all cases. These results point to a lack of correlation between bcl-2 protein expression and the presence of p53 gene mutations. CONCLUSIONS: The level of bcl-2 expression in breast carcinoma is not associated with any somatic abnormality or epigenetic change at the bcl-2 locus. Conversely, although bcl-2 expression is related inversely to p53 protein expression, the analysis of p53 mutations (limited to exons 5-9) failed to demonstrate any relationship between p53 mutations and bcl-2 protein expression.

Expression of inducible nitric oxide synthase in breast cancer correlates with metastatic disease.

Breast cancer is characterized by its ability to metastasize rapidly. Factors that might facilitate this metastatic potential include tumor vascularity. Nitric oxide (NO), a labile compound synthesized by NO synthase (NOS), is a major regulator not only of physiologic vascular tone but also of the abnormal vascularity associated with many tumors. To test whether NOS is expressed in primary breast tumors and whether its expression is associated with the presence of metastasis, we analyzed the expression of the inducible NOS in 22 primary breast tumors, and to investigate its association to other gene products related to the metastatic ability of tumor cells, we correlated the expression of the inducible NOS with the expression of the nm23 protein (the product of the putative antimetastatic gene nm23). We found a very strong correlation between the presence of NOS and axillary lymph node metastasis and between NOS and the absence of nm23 protein. These data suggest that NO synthesis and the resulting increase in blood flow to the tumor play a role in the facilitation of tumor metastasis.

[Primary pulmonary nodular amyloidosis]. / Amiloidosis nodular primaria pulmonar.

Localized nodular pulmonary amyloidosis is a very rare condition. We report the case of a man with an upper left lobe mass measuring 8 cm and a nodule measuring 3 cm at the same site. Both were detected by chance in radiologic images. Neoplasia was suspected and diagnosis was made by thoracotomy and lung biopsy. After two years of follow-up, the patient remains asymptomatic and the radiologic images are stable, with no clinical or analytical evidence of systemic amyloidosis or dysproteinemia.

[Idiopathic calcinosis of the scrotum. Cytohistologic diagnosis in biopsy and puncture samples]. / Calcinosis idiopática del escroto. Diagnóstico citohistológico en biopsia y material de punción.

OBJECTIVE: To report an uncommon case of idiopathic calcinosis of the scrotum. METHODS/RESULTS: A case of idiopathic scrotal calcinosis in a 50-year-old male who presented with multiple hard masses in the scrotum is described. Cytological (fine needle aspiration) and histological (biopsy and resection) analyses of these lesions were performed. CONCLUSIONS: Although the etiopathogenesis of scrotal calcinosis remains unknown, the involvement of a dystropic mechanism has recently been described. The usefulness of fine needle aspiration, a method that has gradually been included in the study of testicular pathology, is underscored.

[Correlation among endoscopic, histological, and microbiological findings in gastroduodenal pathology associated with Helicobacter pylori]. / Correlación entre los hallazgos endoscópicos, histológicos y microbiológicos de la patología gastroduodenal asociada a Helicobacter pylori.

We studied 152 patients referred for endoscopy of the upper gastrointestinal tract. Samples of the gastric mucosa and fundus were taken and processed for microbiological and histological study using conventional methods. Acute lesions to the gastrointestinal mucosa were those most frequently observed in endoscopy (28.3%). The overall prevalence of chronic gastritis was 90.8%. The prevalence of infection by H. pylori was 59.8%. All patients in which gastric ulcer or malignant lesions were observed had some kind of chronic gastritis. Duodenal ulcer was the lesion with the highest prevalence and statistical significance with respect to the existence of H. pylori infection. There was a highly significant correlation between H. pylori gastric colonization and lesions of the diffuse antral type and with gastric activity. The significant correlations among the endoscopic, microbiological and histological findings are discussed.

[Greater sensitivity of the PCR-SSCP technique compared with immunohistochemistry for the detection of exon 5 and 6 mutations of gene 53 in breast cancer]. / Mayor sensibilidad de la técnica de RCP-SSCP frente a la inmunohistoquímica para la detección de mutaciones en los exones 5 y 6 del gen p53 en el cáncer de mama.

BACKGROUND: p53 mutations constitute the most frequent genetic abnormality observed in breast cancer. The molecular study of p53 mutations not only provides prognostic information but also allows a better understanding of tumor biology. Most of the studies on p53 have been done by immunohistochemistry procedures. In this study we compare the sensitivity of immunohistochemistry and molecular genetics methods for the detection of p53 mutations. PATIENTS AND METHODS: Forty tumors obtained from breast cancer patients that underwent modified radical mastectomy were analyzed for p53 mutations at exons 5 to 6 through PCR-single stranted conformational polymorphism (SSCP)-sequencing. Moreover, the tumor expression of p53 protein was analyzed by means of immunohistochemistry. RESULTS: All tumors could be amplified by PCR and analyzed by the SSCP technique. Six tumors (15%) showed and altered electrophoretic mobility. In these cases a mutation was confirmed by gene sequencing. On the other hand, only two of these six tumors were positive in the immunohistochemical analysis. CONCLUSIONS: The results confirm the utility of the PCR-SSCP technique for detection of p53 mutations and suggest to be more sensitive than immunohistochemistry.

Analysis of nm23-H1 expression in breast cancer. Correlation with p53 expression and clinicopathologic findings.

Metastasis is the most frequent cause of death in patients with breast cancer. The nm23-H1 and p53 genes have been involved in the development of breast cancer metastasis. We have analyzed the correlation between the expression of nm23 protein and several established clinicopathologic factors. Our results show that the antimetastatic role of nm23-H1 is not related to the cell proliferative status or tumor grade and that it is not associated with the expression of p53. We also demonstrate a strong inverse relationship between the expression of nm23-H1 protein, lymph node metastasis and vascular invasion. These data support the antimetastatic role of the nm23-H1 gene and suggest that nm23-H1 and p53 genes may be involved in different steps of the metastatic process.

[Male pseudohermaphroditism: perineoscrotal pseudovaginal hypospadias]. / Pseudohermafroditismo masculino: hipospadias pseudovaginal perineoescrotal.

We report on a case of pseudovaginal perineoscrotal hypospadias. The gross and microscopic features of this condition are described. The existence of ambiguous external genitalia without gynecomastia together with a complete inhibition of the müllerian structures is confirmed.

Experimental thioacetamide-induced cirrhosis of the liver.

Hepatic cirrhosis is a complex disease in which several biological, biochemical and chemical alterations are combined, none of these alone being sufficient for diagnosis. The morphological characteristics of the final stages of cirrhosis are well known, but the initial lesions and intermediate stages still have not been fully clarified. An experimental model of hepatic cirrhosis by chronic administration over 30 weeks of thioacetamide (50 mg/kg twice weekly) to female Wistar rats has been produced. In a macroscopic, microscopic and ultrastructural study. The different lesions that appeared were evaluated according to the dose of the toxic agent administered up, until hepatic cirrhosis was finally installed; this was after 60 doses of the toxic agent (30 weeks). Discussion is made of the different types of administration and the doses employed to obtain a suitable survival rate for these cases; in our experiments this was 95%. It has been demonstrated in both human and experimental pathology that once the disease itself has been installed, currently there is no rational or useful treatment for it. A beneficial effect has been demonstrated for certain substances, improving the initial and intermediate lesions, so we conclude by stating that it is necessary to further study the hepatic lesions preceeding cirrhosis. Knowledge of these lesions could form the basis for establishing a useful and rational therapy for such cases.

[Hemangiomatous lesions with lymphangiectasias in the large intestine]. / Lesiones hemangiomatosas con linfangiectasias en intestino grueso.

We present the case of a 16 year-old male who presented intestinal hemorrhages since childhood; he was operated in emergency for microhemangiomas located in sigmoid colon and rectum. This lesion can be considered as a complex vascular hamartoma. We review the literature and discuss the differences between this lesion and angiodysplasia.

[Colorectal cancer: retrospective analysis of 762 cases]. / Cáncer colorrectal: análisis retrospectivo de 762 casos.

A study carried out during 1976-1988 on patients with colorectal cancer revealed an increase in the incidence of this disease of 150%, with a sex ratio of 1:1.17, there being a slight predominance in males. In the patients studied, 3.41% were older than 40. In patients in the 60-70 age group, 63.4% of the cases were diagnosed. The most frequent location of the tumor is the rectum (24.4%) and the least frequent (1.57% and 1.04%, respectively) in the hepatic and splenic angles. This was observed in all the years comprising the study. Adenocarcinoma was the commonest type of tumor, there being a highly significant association (p less than 0.001) between the histological type of cancer and the state of infiltration observed at the time of diagnosis.

[Primary duodenal adenocarcinoma; apropos of a case]. / Adenocarcinoma duodenal primario; a propósito de un caso.

Primary duodenal adenocarcinoma is an infrequent malignant tumor that usually affects persons 60 to 70 years-old. Its clinical manifestations are nonspecific. The diagnosis is suspected on the basis of gastroduodenal contrast radiography and it is generally confirmed by endoscopic biopsy. It is slow to metastasize, so early diagnosis can lead to curative surgical treatment. We present a case seen late, of rapid and fatal course. Statistical data are provided and we comment on some aspects of the classification, clinical manifestations, diagnosis and treatment of these tumors.

[Plasmacytoma of the testis: report of a case]. / Plasmocitoma de testículo: presentación de un caso.

We report on a case of testicular plasmacytoma in a patient with multiple myeloma (MM), who is currently in a state of remission following chemotherapy and radiotherapy. The literature is reviewed, and testicular involvement in plasma cell neoplasms is discussed.

[Effect of (+) cyanidanol-3 on experimental liver cirrhosis induced by carbon tetrachloride]. / Efecto del (+) cianidanol-3 sobre la cirrosis hepática experimental inducida por el tetracloruro de carbono.

Liver cirrhosis is a very common illnesses and currently its treatment remains unpromising. In a research line devoted to studying the hepatotoxicity of certain substances we have evaluated the lesions that appear prior to the occurrence of cirrhosis and hepatocarcinoma in order to test, in different assays liver-protecting substances such as cyanidanol-3 that have been seen to lead to an improvement in morphological alterations in the liver and even to reverse such lesions. The mechanisms of action of this compound are discussed. The authors believe that considerable work remains to be carried out but that some day some liver-protecting substances that yield good experimental results may be applied to clinical situations.