Influence of peritoneal fluid on the expression of angiogenic and proteolytic factors in cultures of endometrial cells from women with endometriosis.

BACKGROUND: Endometriosis, defined as the presence of endometrium outside the uterus, is one of the most frequent benign gynaecological diseases. It has been suggested that both endometrial and peritoneal factors, related to angiogenesis and proteolysis, can be implicated in this disease. The aim of this study was to evaluate the influence of peritoneal fluid on the expression of angiogenic and proteolytic factors in cultures of endometrial cells from women with and without endometriosis. METHODS: Endometrial cells were isolated, cultured and treated with endometriotic or normal peritoneal fluid. Vascular endothelial growth factor-A (VEGF-A), urokinase plasminogen activator (uPA), matrix metalloproteinase-3 (MMP-3) and their inhibitors including thrombospondin-1, plasminogen activator inhibitor-1 and MMP inhibitor type 1 (TIMP-1) mRNA levels were evaluated by quantitative RT-PCR, and protein levels were quantified by ELISA. RESULTS: Peritoneal fluid from women with endometriosis induced an increase in VEGF-A and uPA protein and VEGF-A mRNA and uPA mRNA levels in endometrial cell culture from women with (P < 0.01) and without endometriosis (P < 0.05). The highest levels of VEGF-A and uPA were observed in endometrial cell cultures from patients with endometriosis and treated with peritoneal fluid from women with endometriosis. CONCLUSIONS: Peritoneal fluid from women with endometriosis induced more VEGF and uPA expression in endometrial cell culture from women with endometriosis than did normal peritoneal fluid. Endometrial-peritoneal interactions increased angiogenic and proteolytic factors in endometrial cells, which could contribute to the development of endometriotic lesions.

Clinical variables associated with the presence of inflammatory infiltrates in patients with dilated cardiomyopathy undergoing heart transplantation.

INTRODUCTION: Idiopathic dilated cardiomyopathy (DCM) is, together with ischemic heart disease, the major cause of end-stage heart failure leading to heart transplantation. However, an unknown percentage of patients with this diagnosis has inflammatory foci found in the histopathological study of the explanted heart. This fact suggests an undetected process of acute myocarditis as the cause of cardiac dysfunction. OBJECTIVE: The objective of this study was to identify clinical and echocardiographic variables related to the presence of myocardial infiltrates, as a potential guide to determine which patients should undergo endomyocardial biopsy in DCM. MATERIALS AND METHODS: We retrospectively analyzed 161 patients who underwent heart transplantation with a diagnosis of DCM between 1987 and 2007. The presence of inflammatory infiltrates was considered significant when the histopathological study of tissue blocks from the left ventricle showed 1 or more foci per cm(2) of perivascular or interstitial mononuclear or polymorphonuclear cells, whether or not in the presence of cytolysis. RESULTS: Seventeen patients (11%) had these inflammatory histological findings; of them, 6 (35%) showed preponderance of eosinophils and 7 (41%) showed areas of cytolysis. The DCM group with inflammatory infiltrates showed significant differences in terms of younger age (45 +/- 15 vs 50 +/- 11 years; P < .01) and smaller ventricular diameters (P < .05). Male gender was more frequent in this group, and the patients had a poorer clinical status and greater dependence on inotropic drugs. CONCLUSIONS: Inflammatory infiltrates are frequently present in DCM explanted hearts. Although there are no relevant clinical variables to identify subclinical myocarditis, these patients are younger and have smaller ventricular diameters and poorer functional status at the time of transplantation.

Decreased expression of PAI-2 mRNA and protein in pregnancies complicated with intrauterine fetal growth retardation.

An increase in plasma plasminogen activator inhibitors (PAIs), fundamentally PAI type 2 (PAI-2), has been described in normal pregnancy probably because the placenta is the main source of the high plasma levels of this protein. Although we have previously described plasmatic alterations of these inhibitors in pregnancies complicated with intrauterine fetal growth retardation (IUGR), no reports have been published about placental PAI-2 mRNA expression. In the present study, the placental PAI-2 expression determined in pregnancies complicated with IUGR and in severe preeclamptic patients was compared with that of normal pregnancies in order to identify the placental cell types expressing PAI-2 and to determine whether the production of PAI-2 is altered in placentas from IUGR. In situ hybridization analyses show that the syncytiotrophoblasts are the cells with the greatest PAI-2 expression in placenta. We report that the significant decrease in plasma and placental PAI-2 levels in IUGR groups is fundamentally due to a diminished expression of PAI-2 mRNA in placenta.

Activation of glomerular mesangial cells by hepatocyte growth factor through tyrosine kinase and protein kinase C.

Hepatocyte growth factor (HGF) induces mitogenesis, chemotaxis, and tubule formation in renal epithelial cells. This study examined the effects of wortmannin and protein kinase C (PKC) inhibitors on HGF-mediated changes in metabolic activity in glomerular mesangial cells and renal epithelial carcinoma A498 cells. The extracellular acidification rate of transformed mouse glomerular mesangial cells and A498 cells was measured as an index of metabolic activity with a microphysiometer. HGF increased the acidification rate of mesangial cells and A498 cells in a concentration-dependent fashion that was inhibited completely by the tyrosine kinase inhibitor tyrophostin-23 (100 microM). The PKC inhibitors RO-32-0432 and SKF-57048 also inhibited HGF-induced acidification. The IC50 values for SKF-57048 were 59 +/- 2 and 20 +/- 10 nM in mesangial cells and A498 cells, respectively (P < 0.05). 12-O-Tetradecanoylphorbol 13-acetate (TPA), a phorbol ester that activates PKC, increased acidification in mesangial and epithelial cells similar to HGF. Wortmannin, an inhibitor of phosphatidylinositol (PI) 3-kinase (IC50 value 1-10 nM), inhibited HGF-induced acidification with an IC50 of 93 +/- 31 and 9 +/- 1 nM in mesangial and A498 cells, respectively (P < 0.05). In contrast, there was no significant difference in the IC50 value of wortmannin for epidermal growth factor (EGF)-induced acidification between mesangial and A498 cells (23 +/- 9 vs 14 +/- 1 nM, respectively). Because the IC50 value for wortmannin in inhibiting HGF but not EGF-induced acidification was an order of magnitude higher in mesangial cells than in epithelial A498 cells, a wortmannin-sensitive PI 3-kinase pathway may not be involved in HGF-mediated acidification in mesangial cells.

Successful revascularization of large isolated tracheal segments.

The omentum has been shown to be of use in clinical and experimental revascularization of tracheobronchial anastomoses. We have evaluated the possibility of revascularizing large and completely isolated tracheal segments while preserving the main tracheal characteristics. Ten experiments were performed in dogs, introducing 10-14 cartilage ring tracheal segments enveloped in omentum into the abdomen. Revascularization resulted in all cases with preservation of tracheal consistency. In only two cases were small mucosal necrotic zones observed. The experimental model thus appears to be of use in the revascularization of large tracheal segments, with excellent preservation of both cartilage and mucosa.

Radioimmunometric assay of B-type natriuretic peptide (BNP) in heart transplantation: correlation between BNP determinations and biopsy grading of rejection.

The aim of this study was to determine whether elevated brain natriuretic peptide (BNP) levels after heart transplantation are correlated with the severity of rejection by using endomyocardial biopsy (EMB) and echocardiographic parameters indicative of ventricular function of the transplanted heart. This was an observational study of 80 orthotopic heart transplant recipients (11 women and 69 men; mean age 53+/-11 years). BNP determinations were performed within 48 h of endomyocardial biopsy. The echocardiographic study and BNP determination were also performed in a group of healthy volunteers. We found significantly higher BNP mean levels in heart transplant patients than in healthy volunteers (volunteers, 16.7+/-16.2 pg.ml-1; transplant, 213.4+/-268.6 pg.ml-1; P<0.001). Transplant recipients with rejection grades 2, 3 and 4 on EMB had significantly higher BNP levels than those with rejection grades 0 and 1 (higher rejection grade, 162.5+/-168.4 pg.ml-1; lower rejection grade, 292+/-361.8 pg.ml-1; P<0.01). BNP values of patients with good left ventricular function (LVF) were significantly lower than in patients with mildly and moderately impaired LVF and patients with severely impaired LVF (good function, 199.76+/-233.6 pg.ml-1; mildly/moderately impaired LVF, 937+/-644.5 pg.ml-1; severely impaired LVF, 1038+/-491.2 pg.ml-1; P<0.001). It is concluded that BNP plasma levels are elevated in heart transplant patients compared to the normal population. The distribution of BNP levels in heart transplanted patients show a wide range. BNP elevation is greater in patients with higher rejection grades on EMB and greater impairment of left ventricular function.

Antigens of the major histocompatibility system in ischemic heart disease and idiopathic dilated cardiomyopathy.

BACKGROUND: Although dilated cardiomyopathy (DCM) is a disease of unknown and probably multifactorial etiology, a change in immune mechanisms is presumably significant, with many abnormalities in humoral and cellular responses having been reported. The heart thus becomes the target organ for an initial episode of myocardial damage that triggers an autoimmune response. HYPOTHESIS: The purpose of this study was to analyze the frequency of different human leukocyte antigens in patients with a diagnosis of well-advanced DCM and ischemic heart failure, comparing them with a control group of presumably healthy subjects. METHODS: The group with dilated cardiomyopathy consisted of 50 patients (7 women and 43 men), aged from 14 to 64 years. The group with ischemic heart disease included 76 patients (4 women and 72 men), with ages ranging from 34 to 64. The control group, consisting of 1,337 presumably healthy subjects from the Spanish Mediterranean area, was recruited based on paternity studies. RESULTS: Compared with the control group, we found in DCM a greater incidence of B15 (20 vs. 6%) and DQ3 (83 vs. 50%) antigens. In ischemic heart disease we found a lower incidence of A1 (3 vs. 22%), B8 (5 vs. 12%), and DQ2 (16 vs. 50%) in comparison with the control group. CONCLUSIONS: In the Spanish Mediterranean area, the presence of antigens B-15 and DQ3 would be associated with advanced DCM. The absence of antigens A1, B8, and DQ2 would be associated with the occurrence of severe ischemic heart disease.

[Heart angiosarcoma and heart transplantation. Report of a case]. / Angiosarcoma cardíaco y trasplante cardíaco. A propósito de un caso.

We present the case of a 29-year-old women with a cardiac primary angiosarcoma diagnosis. The initial symptom was a cardiac tamponade. The tests for screening metastasis proved negative. She was preoperatively treated with chemotherapy, followed by a heart transplant. There were no incidents related to surgery nor to the transplant except for a rejection in the second week biopsy. Four weeks after the transplant, the patient had a sudden dyspnea, the radiological tests confirmed the existence of a massive pleural overflow and lung and pleural metastasis. All types of therapeutical approaches were rejected except for pleurodesis. The patient died 60 days after the heart transplant.

[The usefulness of the isovolumetric relaxation time of both ventricles in detecting acute rejection in the heart transplant patient]. / Utilidad del tiempo de relajación isovolumétrica de ambos ventrículos en la detección de rechazo agudo del paciente trasplantado cardíaco.

AIMS: The purpose of our study was to evaluate the usefulness of the isovolumetric relaxation time in both ventricles when diagnosing acute rejection in transplanted patients. METHOD: 68 endomyocardial biopsies were performed on a total of 38 patients. An echocardiographic study was carried out within the first 24 hours of each biopsy. All registrations were made by the same person. The isovolumetric relaxation time was measured in the left and right ventricles. The patients were divided according to two criteria: according to the degree of rejection (0-I, II, III) and according to whether the rejection was treatable (III) or non-treatable (0, I and II). RESULTS: In both ventricles, there was a progressive decrease of the isovolumetric relaxation time corresponding to higher degrees of rejection: in the left ventricle (0-I = 90 +/- 16; II = 74 +/- 16; III = 70 +/- 26; significant differences of II and III in relation to 0-I) as well as in right ventricle (0-I = 43 +/- 16; II = 37 +/- 14; III = 29 +/- 8; significant difference of III in relation to 0-I). The patients with treatable and non-treatable rejection were compared: no differences were found in the isovolumetric relaxation time of the left ventricle (0, I and II = 85 +/- 16 vs III = 70 +/- 26), but they were found in the right ventricle (0, I and II = 41 +/- 15 vs III = 29 +/- 8). CONCLUSIONS: Acute heart rejection induces a decrease of the isovolumetric relaxation time in both the left ventricle and the right ventricle. However, the isovolumetric relaxation time of the right ventricle seems to be a more useful parameter than isovolumetric relaxation time of the left ventricle, as it permits to detect whether an acute heart rejection is treatable or non-treatable.

[The incidence of major histocompatibility system antigens in dilated and ischemic myocardiopathies]. / Incidencia de los antígenos del sistema mayor de histocompatibilidad en la miocardiopatía dilatada e isquémica.

AIM: The purpose of this study was to analyze the frequency of the different antigens of HLA in patients with diagnosis of very advanced dilated cardiomyopathy and ischemic heart disease by comparing them with a control group of supposedly healthy subjects. MATERIAL AND METHOD: The group of dilated cardiomyopathy consisted of 35 patients (8 women and 27 men) aged between 14 and 60 years. The group of ischemic heart disease included 32 patients (4 women and 28 men) aged between 34 and 64 years. The control group comprised 1337 subjects of the Spanish Mediterranean area, supposedly healthy and recruited from paternity studies. RESULTS: In dilated cardiomyopathy we found a higher incidence in comparison with the control group of the A-2 (62.86% vs 46.22%), B-12 (60.00% vs 32.38%) and DQ-3 (82.86 vs 49.96%) antigens, and a lower incidence of B-51 (0.00% vs 12.49%). In ischemic heart disease we found, when comparing to the control group, a higher incidence of A-11 (31.25% vs 13.08%) and A-29 (34.38% vs 14.58%) antigens and a lower incidence of DQ-2 (15.63% vs 49.88%). CONCLUSIONS: In the Spanish Mediterranean area, the presence of A-2, B-12 and DQ-3 antigens, as well as the absence of B-51 would favour the appearance of advanced dilated cardiomyopathy. The presence of the A-11 and A-29 antigens would predispose to ischemic cardiomyopathy while the presence of DQ-2 would have a protective effect on the appearance of this cardiopathy.

[Coronary involvement in Takayasu's arteritis]. / Afectación coronaria en la arteritis de Takayasu.

Takayasu's arteritis is a chronic inflammatory disease that primarily affects young women. Cardiac involvement is infrequent and it includes aortic regurgitation, pericarditis, angor pectoris or myocardial infarction due to coronary narrowing and cardiac heart failure due to coronary involvement and/or high blood pressure. A patient with Takayasu's aortitis and angina pectoris due to severe narrowing of the left coronary arterial ostia is described.

[Preoperative embolization in meningioma of the ponto-cerebellar angle. Indications and advantages]. / Embolización preoperatoria en un meningioma del ángulo ponto-cerebeloso. Indicaciones y ventajas.

Preoperative embolization of meningiomas has been performed in order to reduce surgical hemorrhage during the removal of these vascularized tumors. In this paper we emphasize the fact that occlusion of the tumoral vessels by artificial emboli produces an ischemic necrosis that greatly helps tumor exeresis. This is especially useful in meningiomas of certain localizations requiring complex surgical approaches. In our case, a giant ponto-cerebellar meningioma was dried up totally and then removed with relative ease through a conventional suboccipítal lateral craniectomy. The technique, indications and control of preoperative embolization are reviewed.

[Pregnancy, breast cancer and tumor infarction]. / Embarazo, cáncer de mama e infartación tumoral.

Breast cancer is infrequently associated with pregnancy. However, such cases often pose diagnostic and treatment problems. The infarction of breast lesions is likewise uncommon, and generally hinders intraoperative morphologic diagnosis. A 33-year-old woman in the 37th week of pregnancy presented with a breast tumoration of progressively increasing size and associating mastalgia. Following fine-needle aspiration and trut-cut biopsy with posterior amplified tumorectomy and axillary dissection, a ductal neoplasm was identified with an infarction affecting 90% of the tumoral surface. The where no axillary adenopathies, and the patient was estrogen receptor-negative. An analysis is made of this rare association of pregnancy and infarcted breast cancer, with an evaluation of the role of fine-needle aspiration and/or gestation may play in the occurrence of tumor infarction.

[Literature review on the stability of diluted mixtures of cytostatic agents]. / Revisión bibliográfica de la estabilidad de las mezclas diluidas de citostáticos.

Up-to-date, evidence-based consensus protocols are an increasingly incorporated tool in health care. These protocols, clinical pathways, etc., represent a major support of health-care quality, namely scientific-technical quality. Compliance with these protocols by all team members guarantees that all patients be provided with an adequate level of health-care quality in the light of current knowledge and using available means. This principle of uniformity and quality in health care is essential for all, no matter the level of health care delivered or the activity being developed. In the Pharmacy Department and, more specifically, in the Unit of Cytostatic Agent Reconstitution and Dosing, knowledge and consensus on stability conditions and timing for diluted mixtures are essential to reach area-related quality standards. From literature references that are most relevant to or most widely used by in-hospital pharmacy departments, we designed a documented stabilities protocol to be used as a tool to: Augment preparation quality by including a documented expiry date within labels, optimize management on the basis of scientific criteria for mixtures not administered to patients and returned to the Pharmacy Department and design alternatives to hospitalization and outpatient delivery programs to improve end-user satisfaction and, therefore, health-care quality.

[Assessment of prescription in discharge reports at a university hospital]. / Evaluación de la prescripción de medicamentos en los informes de alta en un hospital universitario.

OBJECTIVE: To determine and analyze drug prescription at hospital discharge, mainly regarding generic drug use, use of novel but irrelevant therapeutic agents and use of low therapeutic value drugs (LTVD). MATERIAL AND METHODS: In a retrospective study 195 discharge reports from 11 different departments in a 450-bed general hospital were analyzed for a monthly period. An Access database allowed us to record the number of prescriptions, each drugs therapeutic group according to the Anatomical Therapeutic Chemical (ATC) Classification System, prescribed drugs for which generics are available, prescribed C-group drugs, LTVD drugs, etc. RESULTS: Following an analysis of results, only 6.17% of all drugs were prescribed according to their generic name, when this would have been possible in 22.8%. If only the most efficient agents had been prescribed, savings would have amounted to 589.3 Euros. In all, 1.28% of prescribed drugs were LTVD, and 1.15% had irrelevant therapeutic value. CONCLUSION: Although specialized medical prescription represents a minimum of total prescriptions in a healthcare area, measures intended to improve quality will have a positive impact on primary care prescriptions. These measures include information to physicians on more efficient preparations, plus the design of a Pharmacotherapeutic Guide to unify pharmacologic criteria in the Area.

Expression of angiogenic factors in endometriosis: relationship to fibrinolytic and metalloproteinase systems.

BACKGROUND: Endometriosis is a highly prevalent, benign disease in which the angiogenic, fibrinolytic and metalloproteinase (MMP) systems may be implicated. The objective of this study is to analyse mRNA expression and protein levels of several angiogenic factors and to correlate them with several components of the fibrinolytic and MMP systems in samples from 71 women with endometriosis and 50 controls. METHODS AND RESULTS: Eutopic endometrium showed higher mRNA expression of vascular endothelial growth factor (VEGF) in patients than in controls. However, ovarian endometrioma had lower VEGF mRNA levels than did the eutopic endometrium of patients. Similar results were obtained for VEGF protein levels. On the other hand, a significant increase in thrombospondin-1 (TSP-1) levels was observed in ovarian endometrioma than in eutopic endometrium. The peritoneal fluid from women with endometriosis showed a significant increase in VEGF, urokinase-type plasminogen activator (uPA) and MMP-3 levels than that of controls. A significant correlation was observed between the levels of VEGF and uPA in endometrium and in peritoneal fluid. CONCLUSIONS: Endometrium and peritoneal fluid from women with endometriosis have increased levels of VEGF, uPA and MMP-3 levels. Therefore, the development of endometriotic implants at ectopic sites may be facilitated, promoting the progress of the endometriosis.

mRNA analysis of several components of the plasminogen activator and matrix metalloproteinase systems in endometriosis using a real-time quantitative RT-PCR assay.

BACKGROUND: The plasminogen activator (PA) and matrix metalloproteinase (MMP) systems are implicated in the establishment of endometriosis. The mechanisms by which these systems are involved in the pathogenesis of this disease are not well defined and controversial results have been published. The aim of this study was to analyse mRNA and protein levels of several components of the PA and MMP systems in endometriotic tissue and endometrium from women with and without endometriosis. METHODS AND RESULTS: Real-time quantitative RT-PCR assays were developed to quantify mRNA levels of these components in 57 women with endometriosis and 32 controls. Endometrium of women with endometriosis showed higher mRNA and antigenic levels of urokinase type-PA (uPA) and MMP-3 than endometrium from controls. In these patients, ovarian endometriotic tissue had higher mRNA and antigenic levels of PA inhibitor type 1 (PAI-1) and MMP inhibitor type 1 (TIMP-1) than endometrium. CONCLUSIONS: The increase in mRNA and protein levels of uPA and MMP-3 observed in endometrium of women with endometriosis may facilitate the attachment of endometrial tissue to the peritoneum and ovarian surface, as well as the invasion of the extracellular matrix. This process would lead to the formation of early endometriotic lesions. Once the ovarian endometriotic cyst is developed, PAI-1 and TIMP-1 would increase which could explain the frequent clinical finding of an endometrioma without invasion of the adjacent ovarian tissue.

The role of maternal alcohol damage on ethanol teratogenicity in the rat.

To study the severity and degree of in utero alcohol effects in relation to the rate of maternal alcohol damage, multiparous 1-year alcohol-fed rats were used, with an appropriate pair-fed control group. During pregnancy, alcoholic dams showed relatively high acetaldehyde levels (41 +/- 19 mumol/l) and blood alcohol levels of 22.8 +/- 14 mmol/l. They also showed marked histological alterations in liver as well as high serum aspartate-aminotransferase, alanine-aminotransferase, alkaline phosphatase, glutamate dehydrogenase, and gamma-glutamyltransferase activities. The increase in serum enzyme levels did not correlate with an increase in hepatic enzyme levels since only glutamate dehydrogenase was enhanced in liver after 1 year of alcohol intake. In addition, except for an increase in low Km aldehyde dehydrogenase activity, there were no changes in liver alcohol metabolizing enzymes in chronic alcohol vs. pair-fed females. Alcoholic rats showed a high incidence of damage in their progeny (resorptions, immature fetuses, decrease in fetal weight, etc.), and rats with the highest serum levels of the above enzymes (especially glutamate dehydrogenase and gamma-glutamyl transferase) had severely affected progeny. Rats with minimal histological liver damage, in contrast, did not show resorptions. Thus, the results presented suggest that the stage of maternal alcohol illness, as indicated mainly by the extent of liver damage, plays an important role in the frequency and severity of in utero alcohol effects in the rat.

Unusual presentation of non-Hodgkin lymphoma of the spleen.

We report a patient with highly malignant primary centroblastic lymphoma of the spleen (diffuse, large non-cleaved cell type). The tumor presented as splenomegaly due to a large mass containing diffuse punctuate calcifications. The latter have not been previously described as a radiological finding in this neoplasm.