Impact of serpentinized peridotite mine waste on the composition and quality of sediments in the Ría de Ortigueira (Galicia, NW Spain).

The Ría de Ortigueira is an environmentally well conserved; however, the sediments show high concentrations of toxic elements. In some zones, the concentrations of Ni (60-1080 mg kg-1) and Cr (9-567 mg kg-1) were extremely high, while the concentrations of other toxic elements were within normal ranges. PCA revealed that metal enrichment was due to dumping of waste sludge from a peridotite mine. The study of marine currents showed that the exit of the contaminated waste towards the external zone is restricted by the low energy of the residual currents, and the sludge therefore remains trapped in the internal zones. The potential ecological risk was moderate for all areas of the ría, reaching high values close to the mouth of the river Landoi. Finally, geochemical fractioning showed that most of the metals are associated with Fe oxyhydroxides which can become unstable and release adsorbed or coprecipitated metals, especially Ni.

Consenso de VIH: tratamiento antirretroviral en adultos

Consenso para el tratamiento antirretroviral en adultos

Antiretroviral Adult treatment Consensus

Consenso de VIH-2: tratamiento antirretroviral en niños, adolescentes y embarazadas

Consenso para el tratamiento antirretroviral de las mujeres embarazadas, los adolescentes y los niños

Consensus of Antiretroviral treatment of pregnant women, adolescents and children

Consenso de VIH-3: adherencia al tratamiento antirretroviral y consideraciones en situaciones especiales

Consenso acerca de la adherencia al tratamiento antirretroviral en situaciones especiales

Consensus about the antiretroviral treatment adherence in special situations

HRTEM Microstructural Characterization of ß-WO3 Thin Films Deposited by Reactive RF Magnetron Sputtering.

Though tungsten trioxide (WO3) in bulk, nanosphere, and thin film samples has been extensively studied, few studies have been dedicated to the crystallographic structure of WO3 thin films. In this work, the evolution from amorphous WO3 thin films to crystalline WO3 thin films is discussed. WO3 thin films were fabricated on silicon substrates (Si/SiO2) by RF reactive magnetron sputtering. Once a thin film was deposited, two successive annealing treatments were made: an initial annealing at 400 °C for 6 h was followed by a second annealing at 350 °C for 1 h. Film characterization was carried out by X-ray diffraction (XRD), high-resolution electron transmission microscopy (HRTEM), scanning electron microscopy (SEM), and atomic force microscopy (AFM) techniques. The ß-WO3 final phase grew in form of columnar crystals and its growth plane was determined by HRTEM.

Antígenos HLA de clase ii en pacientes con artritis reumatoide ecuatorianos y asociación con el factor reumatoide, anticuerpos anti-citrulinados y nódulos reumatoides / HLA antigens class ii in patients with ecuadorian rheumatoid arthritis and association with the rheumatoid factor, anti-citrulinated antibodies and rheumatoid nodules

Objetivo Determinar los antígenos de Clase II en 52 pacientes con artritis reumatoide (AR) y 52 controles, y establecer la asociación de los antígenos de Clase II DRB1* con el factor reumatoide (FR), anticuerpos anti-péptido cíclico citrulinados (anti-CCP) y nódulos reumatoides. Material y Métodos Estudio de casos y controles, realizado en pacientes del Hospital Vicente Corral Moscoso y José Carrasco Arteaga de la ciudad de Cuenca-Ecuador; se incluyeron 52 pacientes con AR que cumplen los criterios del Colegio Americano de Reumatología (ACR) y 52 controles. Se identificaron a través de la Reacción en Cadena de la Polimerasa (PCR), los diferentes tipos de HLA que tienen cada uno de los 52 pacientes con AR y controles. Se buscó asociación de los HLA-DR1* con la AR y las pruebas de laboratorio: FR, anticuerpos anti-CCP y con nódulos reumatoides. El análisis estadístico se realizó con el cálculo de OR (IC 95%) y 2 de Pearson con corrección de Fisher y la información se procesó en SPSS v15. Resultados Los alelos más frecuentes fueron HLA-DRB1*09 (17,3%) OR 3,42 (IC95% 1,21 ­ 10,14) y HLADRB1*04 (32,7%) OR 1,81 (IC95% 0,93 ­ 3,55). Tanto el factor reumatoide como los anticuerpos anti-péptido cíclico citrulinados estuvieron presentes en el 82,7% de pacientes con artritis reumatoide y los nódulos reumatoides se detectaron en el 19,2% de pacientes con AR. Se encontró asociación entre los nódulos reumatoides con valores altos de FR y anti-CCP y estos a su vez con los alelos HLA-DRB1*14 y HLADRB1*04. La mayoría de pacientes con alelos HLA-DRB1*09 con predominio heterocigoto tuvieron anticuerpos anti-CCP y FR positivo. Conclusión La presencia de los alelos HLA-DRB1*04 y HLADRB1*09 está relacionada con la susceptibilidad de presentar AR en pacientes ecuatorianos, además se relaciona con valores elevados de factor reumatoide y anticuerpos anti-péptido cíclico citrulinados.

Objective To determine Class II antigens in 52 patients with rheumatoid arthritis (RA) and 52 controls and establish the association of Class II antigens DRB1* with rheumatoid anti- cyclic citrullinated peptide (anti-CCP) rheumatoid factor (RF), and antibodies rheumatoid nodules. Methods and Materials Case and control study in patients from the Hospital Vicente Corral Moscoso and José Carrasco Arteaga of Cuenca-Ecuador; 52 patients with rheumatoid arthritis who satisfy the criteria of ACR-American College of Rheumatology and 52 healthy patients. The different types of HLA from each 52 patients with rheumatoid arthritis and controls were identified through PCR - Polymerase Chain Reaction. Association of HLA-DR1* with rheumatoid arthritis and rheumatoid factor, anti-citrullinated and rheumatoid nodules. The statistical analysis was done by the OR (IC 95%) and 2 of Pearson with Fisher correction and the information was processed in SPSS v15. Results The most frequency alleles were HLA-DRB1*09 (17,3%) OR 3,42 (IC95% 1,21 ­ 10,14) and HLADRB1*04 (32,7%) OR 1,81 (IC95% 0,93 ­ 3,55). The rheumatoid factor and anti-cyclic citrullinated peptide antibodies were presented in 82,7% of patients with rheumatoid arthritis and rheumatoid nodules was detected in the 19.2% of patients with rheumatoid arthritis . An association between rheumatoid nodules with high values of rheumatoid factor and anti-ccp and these in turn with HLA-DRB1*14 y HLA-DRB1*04 alleles were found. The majority of patients with HLA-DRB1*09 alleles with heterozygote predomination had positive anti-ccp and rheumatoid factor. Conclusion The presence of HLA-DRB1*04 and HLA-DRB1*09 alleles is related with the susceptibility of present rheumatoid arthritis in Ecuadorian patients, and it is related with higher values of rheumatoid factor and anti-cyclic citrullinated peptide.

Infecciones urinarias adquiridas en la comunidad: epidemiología a los antimicrobianos y opciones terapéuticas / Urinary infections acquired in community: epidemiology resistance to antibiotics and therapeutic options

Las infecciones del tracto urinario (ITU) constituyen uno de los principales motivos de consulta en el ámbito de atención primaria. En los últimos años se han producido cambios sustanciales en los patrones de sensibilidad de los principales patógenos urinarios, lo que ha condicionado cambios en el tratamiento empírico de éstas. El objetivo de este trabajo fue caracterizar las ITU en pacientes de la comunidad. La muestra estuvo conformada por 71 pacientes de ambos sexos mayores de 18 años. El género más afectado fue el femenino (80,28%). Los signos y síntomas más frecuentes fueron dolor lumbar, disuria y dolor abdominal. La presencia de cálculos renales fue el más importante factor predisponente (39,43%), seguido de la menopausia (23,94%). El 63,38% de los pacientes presentaron ITU previas. Escherichia coli fue el agente causal más frecuente (63,89%), seguido de Proteus mirabilis (6,94%). Las enterobacterias aisladas presentaron elevados niveles de resistencia a ampicilina, cefalotina y norfloxacina. El 51,52% de las enterobacterias presentaron resistencia a las fluoroquinolonas y en el 16,67% se demostró la producción de ß-lactamasas de espectro expandido (BLEE). La mayoría de los agentes causales fueron sensibles a la nitrofurantoína independientemente de la presencia de mecanismos de resistencia que afectan a otros grupos de antimicrobianos

Urinary tract infections (UTI) are one of the main reasons for consultation in primary care. In recent years, there have been substantial changes in susceptibility patterns for major urinary tract pathogens, which have conditioned changes in their empirical treatment. The aim of this study was to characterize UTI in patients from the community. The sample consisted of 71 patients of both sexes over 18 years of age. The most affected sex was the female (80.28%). The signs and symptoms were lumbar pain, dysuria and abdominal pain. The presence of kidney stones was the most important predisposing factor (39.43%), followed by menopause (23.94%); 63.38% of the patients had a previous UTI. Escherichia coli was the most common agent (63.89%), followed by Proteus mirabilis (6.94%). Isolates showed high levels of resistance to ampicillin, cephalothin and norfloxacin. 51.52% of the Enterobacteriaceae were resistant to fluoroquinolones and 16.67% showed production of extended spectrum ß-lactamases (ESBL). Most of the agents were susceptible to nitrofurantoin, regardless of the presence of resistance mechanisms affecting other antibiotic groups

Reflujo gastroesofágico en pediatría / Gastro esophageal reflux disease in pediatrics

Gastro Esophageal Reflux (GER) is the passage of gastric content to the esophagus with or without vomitus or regurgitation. GER is a physiological process that occurs in 50 percent of newborns, spontaneously resolving at 12 to 14 months of age. When this retrograde gastric content reaches the esophagus and produces uncomfortable symptoms or complications, a disease is produced, named Gastro Esophageal Reflux Disease or GERD. Reflux episodes occur due to transient relaxation of the inferior esophagic sphincter, triggered by distension of the gastric fundus. Nursing children show higher frequency of episodes of relaxation due to a higher frequency of feeding than older children do. Most frequent symptoms include frequent regurgitation, with out without vomit, weight loss or poor weight gain, irritability, pyrosis, hematemesis, dysphagia, odinophagia, sibillances, stridor, cough, dysphonia, among others. These are very inespecific symptoms. Studies for GERD include 24 H pHmeasurement, which permits the study of associatrion between one of these symptoms and reflux, as well as the study of those patients with poor response to medical treatment. Unfortunately, it does not detect non-acid reflux. Multichannel intraluminal pH impedance measurement is a relatively new exam that measures the movement of fluids, gas or solids by a change in electric impedance through the esophagus. It allows to detect small volumes, type of content, acid and non acid and correlated with symptoms of GERD. Other useful exams include endoscopy with biopsy of the esophagus for diagnosis of esophagitis, esophageal mannometry for specific cases and gastroesophagic scintigram for pulmonary aspiration. Treatment include change in lifestyle (feeding, position, habits), pharmacological treatment and surgery. Medical treatment includes antacids, PPIs, most frequently omeprazole, H2 blockers such as ranitidine have a lower rate of healing of esophagitis than omeprazole, and appears to show tachyph...

El reflujo gastroesofagico (RGE) es el paso del contenido gástrico hacia el esófago con o sin vómitos o regurgitación. El RGE es un proceso normal fisiológico que ocurre en el 50 por ciento de los lactantes, resolviéndose espontáneamente a los 12 a 14 meses de edad. Cuando este contenido retrógrado gástrico alcanza al esófago y produce síntomas molestos o complicaciones se produce la enfermedad por RGE o ERGE. Los episodios de reflujo se producen principalmente por relajaciones transitorias del esfínter esofágico inferior (LES) gatillado por distensión del fondo gástrico. Los lactantes presentan mayor numero de episodios de relajación del LES debido a que se alimentan con más frecuencia que los niños mayores. Los síntomas más frecuentes asociados con reflujo patológico son regurgitaciones frecuentes con o sin vómitos, pérdida de peso o pobre ganancia de peso, irritabilidad, pirosis, hematemesis, disfagia, odinofagia, sibilancias, estridor, tos, disfonía entre otros, estos síntomas son poco específicos en lactantes y niños menores. Los exámenes para el estudio de ERGE son la pH metría de 24 hrs que permite estudiar la asociación entre un síntoma y el reflujo, también es útil para estudiar aquellos pacientes con mala respuesta al tratamiento médico, sin embargo, tiene el inconveniente que no detecta episodios de reflujo no ácidos. La pH-Impedanciometria Intraluminal multicanal (pH IIM) es un examen relativamente nuevo mide los movimientos de fluidos, gas o sólidos a través de cambios en la impe-dancia eléctrica a lo largo del esófago; permite distinguir pequeños volúmenes refluidos y el tipo de contenido, detecta reflujo acido y no acido y correlaciona síntomas con ERGE. Otros exámenes útiles son la endoscopia con biopsia de esófago para estudio de esofagitis. La Manometría esofágica tiene sus indicaciones específicas y la Cintigrafía gastroesofágica que es otro examen que se utiliza principalmente para estudiar aspiración pulmonar. El tratamiento incluye cambi...

Metaloenzimas tipo VIM detectadas en aislamientos clínicos en Pseudomonas aeruginosa en cuatro hospitales en Venezuela / VIM- type metalloenzymes detected in clinical isolates of Pseudomonas aeruginosa in four Venezuelan hospitals

En los últimos años se ha observado mundialmente un incremento del aislamiento de cepas de Pseudomonas aeruginosa resistentes a múltiples antibióticos, siendo la producción de β-lactamasas la principal causa de resistencia a los antibióticos β-lactámicos, los cuales constituyen la única opción terapéutica en muchos casos. Las metalo-β-lactamasas (MBLs) son una familia de enzimas degradadoras de β-lactámicos que recientemente han emergido como determinantes de resistencia de importancia clínica y que son activas contra los carbapenem; no hidrolizan a los monobactámicos; son inhibidas por agentes quelantes de iones metálicos como el EDTA y el ácido dipicolínico; no son inhibidas por el ácido clavulánico, sulbactan ni tazobactan y presentan uno o dos iones de zinc en su sitio activo. Este reporte describe la detección de MBLs tipo VIM mediante ensayos fenotípicos y moleculares en nueve cepas de Pseudomonas aeruginosa resistentes a carbapenems aisladas de muestras clínicas de cuatro hospitales de Venezuela. Por métodos fenotípicos se evidenció que 100% de las cepas eran productoras de MBLs, y por PCR todas las MBLs resultaron ser de la familia VIM, las cuales confieren alto nivel de resistencia a los antibióticos β-lactámicos, con excepción del Aztreonam, único antibiótico al cual se observó sensibilidad, complicando así las opciones terapéuticas.

In recent years there has been a worldwide raise in the isolation of Pseudomonas aeruginosa strains with resistance to multiple antibiotics, being β-lactamase production the main cause of resistance to β-lactamic antibiotic (which are the only therapeutic option in many cases). The MBLs are an β-lactamic- degrading enzymatic family that have emerged as clinically relevant resistance determinants and are active against carbapemens, don´t hydrolise monobactams, have one or two zinc ions in its active site and are inhibited by metallic ions chelating agents such as EDTA and dipicolinic acid, but aren´t inhibited by β-lactamase inhibitors such as clavulanic acid, sulbactam or tazobactam. This report describes the detection of VIM-type MBLs by phenotypic and molecular methods in 9 carbapenemsresistant Pseudomonas aeruginosa strains isolated from clinical samples of four Venezuelan hospitals. By phenotypic methods it was evidenced that 100% of the strains were MBLs producers, and by final- point PCR it was determined that all the MBLs were from the VIM family, which confer high- level resistance to the β-lactamic antibiotics (except Aztreonam), and because they are carried by plasmids with the ability to transfer horizontally to other bacterial families, they can be responsible for therapeutic complications in an individual or the patients collective.

Expression of gp120 as surrogate marker in HIV infection / Expresión de gp120 como marcador de progresión en la infección por VIH

Ninety seven patients infected with HIV -1 were studied to evaluate the expression of gp120 in peripheral blood CD4+ T lymphocytes as a surrogate marker of disease progression, analyzing correlation of it with viral load, T cell subsets, expression of activation markers, HAAR T and clinical signs and symptoms. In patients who had not received any antiretroviral therapy (naive), expression of gp120 had a positive correlation with expression of CD95 in CD4+ T lymphocytes (p=0.0409) and a negative correlation with CD4+ T lymphocytes percentage (p=0.0183) and absolute values (p=0.0165), and with percentage of naüve CD4+ T cells (p=0.0126). In patients under highly active antiretroviral therapy (HAAR T) those correlations were not present. Expression of gp 120 had good correlations with some known surrogate markers of disease progression in naüve patients, but its low level of expression and the fact that those correlations disappear when HAAR T is initiated, limit the use of it as an independent surrogate marker.

Se estudiaron 97 pacientes infectados con VIH-1 para determinar gp120 en células TCD4+ de sangre periférica como un marcador de progresión de enfermedad y su correlación con carga viral, subpoblaciones de células T, marcadores de activación, terapia antiretroviral, síntomas y signos clínicos. En los pacientes quienes no habían recibido terapia antiretroviral (vírgenes), la expresión de gp120 tenía una correlación positiva con la expresión de CD95 en los linfocitos TCD4+: (p=0,0409) y una correlación negativa con el porcentaje de linfocitos TCD4+ (p=0,0183) con los valores absolutos (p=0.0165) y con el porcentaje de células TCD4+ vírgenes (p=0.0126). En los pacientes con alta terapia antiretroviral estas correlaciones no estaban presentes. La expresión de gp120 tuvo buena correlación con algunos marcadores de progresión de enfermedad en pacientes vírgenes, pero su bajo nivel de expresión y el hecho de que esta correlación desaparece cuando la terapia antiretroviral es iniciada, limita su uso como marcador independiente de progresión.

Blockage of angiotensin II type I receptor decreases the synthesis of growth factors and induces apoptosis in C6 cultured cells and C6 rat glioma.

Angiotensin II (Ang II) is a main effector peptide in the renin-angiotensin system and participates in the regulation of vascular tone. It also has a role in the expression of growth factors that induce neovascularisation which is closely associated to the growth of malignant gliomas. We have shown that the selective blockage of the AT1 receptor of angiotensin inhibits tumour growth, cell proliferation and angiogenesis of C6 rat glioma. The aim of this study was to study the effects of the blockage of AT1 receptor on the synthesis of growth factors, and in the genesis of apoptosis in cultured C6 glioma cells and in rats with C6 glioma. Administration of losartan at doses of 40 or 80 mg kg(-1) to rats with C6 glioma significantly decreased tumoral volume and production of platelet-derived growth factor, vascular endothelial growth factor and basic fibroblast growth factor. It also induced apoptosis in a dose-dependent manner. Administration of Ang II increased cell proliferation of cultured C6 cells which decreased by the administration of losartan. Our results suggest that the selective blockage of AT1 diminishes tumoral growth through inhibition of growth factors and promotion of apoptosis.

PCR detection of specific Leishmania-DNA in patients with periodontal disease.

This study deals with the detection of Leishmania braziliensis DNA in gingival specimens from 10 individuals who all had suffered from cutaneous leishmaniasis 5-10 years prior to the examination and all had been treated with anti-leishmaniasis drugs. This preliminary study gives an interesting contribution to the oral microbiology of this disease, with the observation that inflamed periodontal tissues can serve as a factor affecting the dispersion of Leishmania parasites in individuals who had suffered from cutaneous leishmaniasis. These finding are corroborated by the results of polymerase chain reaction (PCR) which demonstrated the presence of Leishmania DNA in tissue samples of patients with periodontal diseases.

Paradoxical effect of aspirin on the growth of C6 rat glioma and on time of development of ENU-induced tumors of the nervous system.

PURPOSE: Administration of acetylsalicylic acid (ASA), an inhibitor of the synthesis of prostaglandins and thrombzoxanes, decreases the incidence of colorectal cancer and other neoplasms and inhibits in vitro some tumor growth. We studied the effect of various doses of ASA on the growth of C6 glioma implanted in rats as well as the effect of chronic administration of ASA on time of development and incidence of tumors of the central nervous system (CNS) induced by prenatal exposure to ethylnitrosourea (ENU). METHODS: In a controlled study, various doses of ASA, 12.5, 25, 50, 100, 200, 300, and 400 mg/kg per day, were administered to Wistar rats in whom a subcutaneous C6 glioma had been transplanted. Changes in tumor size, histologic characteristics, mitotic index, cell proliferation, and vascular density were studied. In a parallel experiment, we administered ASA (70 mg/kg per day) to rats who were prenatally exposed to ENU; treatment started on day 50 of age, and continued until the end of the experiment at day 400. The time of tumor development as well as incidence, localization, and histological diagnosis were compared with matched controls. RESULTS: A paradoxical effect of ASA administration was observed on the dynamics of cell proliferation of C6 glioma. When high ASA doses were administered (200 or 400 mg/kg per day), tumor volume, cell proliferation, vascular density, and mitotic index increased. In contrast, when low doses were administered (12.5 or 25 mg/kg per day) the tumor size diminished. In the second experiment, localization and incidence of CNS tumors induced by ENU were similar in animals treated with ASA and in controls; however, in rats treated with ASA the time of tumor development was shortened. CONCLUSIONS: The growth-promoting effects of high doses of ASA found in the present study in both transplanted and chemically-induced brain tumors, might be due to the blockage of autocrine inhibitory factors dependent on the cyclooxygenase pathway or by increased vascular permeability and blood supply to the tumor due to inhibition of platelet aggregation. In contrast, the inhibition of tumor growth obtained with low ASA doses in transplanted glioma might be due to different mechanisms such as the induction of apoptosis.

Detection and significance of inapparent infection in Chagas disease in western Venezuela.

Inapparent infections of Trypanosoma cruzi were detected in symptomless seropositive people living in close proximity, and under the same conditions of risk, to patients with acute Chagas disease. Similar infections were also detected in sera samples of people from 25 villages of western Venezuela where Chagas disease is endemic. Seropositivity in all the 1,251 studied samples was established by use of 3 serological methods (direct agglutination test, indirect immunofluorescence antibody test, and enzyme-linked immunosorbent assay). Each seropositive sample was tested for detection of anti-T. cruzi-specific immunoglobulin (Ig) M and IgG levels and specific T. cruzi infection by molecular methodology (polymerase chain reaction assay). The combined analysis of the serologic (IgM and IgG levels), molecular (specific T. cruzi DNA), and statistical findings demonstrated the existence of a different stage of T. cruzi infection in asymptomatic patients, which is suggested to be recognized as inapparent infection. Its definition, significance, and comparison with typical Chagas disease phases are presented, and its potential epidemiological importance is discussed.

Green fluorescent protein-tagged Leishmania in phlebotomine sand flies.

In this work we have used for the first time green fluorescent protein (GFP) tagged cells of the human parasite Leishmania donovani to observe its development in the gut of phlebotomine sand flies. Low numbers of GFP-tagged L. donovani were more easily detected than nontagged Leishmania, suggesting that GFP-tagged Leishmania could be used to efficiently study the biology of Leishmania in their vectors, and open the possibility of using nonaxenic flies. Using this method, we found that GFP-tagged L. donovani, the ethiological agent of Old World Kala-azar, were able to establish an infection within the gut of Lutzomyia species, which are vectors of New World Leishmania. The GFP-tagged parasites divide successfully in the gut of colonized and in wild caught Lu. longipalpis (Lutz & Neiva, 1912), Lu. ovallesis (Ortiz, 1952), and Lu. youngi (Feliciangeli & Murillo, 1985). In the case of Lulongipalpis the labeled parasite exhibited a normal anterior development as the one observed in its natural vector.

Myocardial parasite persistence in chronic chagasic patients.

The persistence of Trypanosoma cruzi tissue forms was detected in the myocardium of seropositive individuals clinically diagnosed as chronic chagasic patients following endomyocardial biopsies (EMBs) processed by immunohistochemical (peroxidase-anti-peroxidase [PAP] staining) and molecular (polymerase chain reaction [PCR]) techniques. An indirect immunofluorescent technique revealed antigenic deposits in the cardiac tissue in 24 (88.9%) of 27 patients. Persistent T. cruzi amastigotes were detected by PAP staining in the myocardium of 22 (84.6%) of 26 patients. This finding was confirmed with a PCR assay specific for T. cruzi in 21 (91.3%) of 23 biopsy specimens from the same patients. Statistical analysis revealed substantial agreement between PCR and PAP techniques (k = 0.68) and the PCR and any serologic test (k = 0.77). The histopathologic study of EMB specimens from these patients revealed necrosis, inflammatory infiltrates, and fibrosis, and made it possible to detect heart abnormalities not detected by electrocardiogram and/or cineventriculogram. These indications of myocarditis were supported by the detection of T. cruzi amastigotes by the PAP technique or its genome by PCR. They suggest that although the number of parasites is low in patients with chronic Chagas' disease, their potential for heart damage may be comparable with those present during the acute phase. The urgent necessity for testing new drugs with long-term effects on T. cruzi is discussed in the context of the present results.

Acute Chagas' disease in western Venezuela: a clinical, seroparasitologic, and epidemiologic study.

A clinical, parasitologic, and serologic study carried out between 1988 and 1996 on 59 acute-phase patients in areas of western Venezuela where Chagas' disease is endemic showed 19 symptomatic patterns or groups of symptoms appearing in combination with different frequencies. The symptomatic pattern with the highest frequency was that showing simultaneously fever, myalgia, headache, and Romaña's sign, which was detected in 20% of the acute-phase patients. Asymptomatic individuals and patients with fever as the only sign of the disease made up 15% and 11.9% of the total acute cases, respectively. Statistical correlation analysis revealed that xenodiagnosis and hemoculture were the most reliable and concordant of the five parasitologic methods used; these two methods also showed the highest proportions in detecting any clinical symptomatic pattern in acute-phase patients. A similar high reliability and concordance was obtained with a direct agglutination test, an indirect immunofluorescent antibody test, and an ELISA as serologic tests, which also showed a higher proportion of positive detection of clinical patterns than parasitologic methods (P < 0.001). It is recommended that individuals coming from endemic areas showing mild and/or severe clinical manifestations should be suspected of being in contact or having been in contact with Trypanosoma cruzi, be referred for parasitologic and serologic evaluations to confirm the presumptive clinical diagnosis of acute Chagas' disease, and start specific treatment. The epidemiologic implications of the present findings are discussed and the use of similar methodology to evaluate other areas where Chagas' disease is endemic is suggested.

Evaluation of DNA recombinant methodologies for the diagnosis of Plasmodium falciparum and their comparison with the microscopy assay.

Since 1984, DNA tests based on the highly repeated subtelomeric sequences of Plasmodium falciparum (rep 20) have been frequently used in malaria diagnosis. Rep 20 is very specific for this parasite, and is made of 21 bp units, organized in repeated blocks with direct and inverted orientation. Based in this particular organization, we selected a unique consensus oligonucleotide (pf-21) to drive a PCR reaction coupled to hybridization to non-radioactive labeled probes. The pf-21 unique oligo PCR (pf-21-I) assay produced DNA amplification fingerprints when was applied on purified P. falciparum DNA samples (Brazil and Colombia), as well as in patient's blood samples from a large area of Venezuela. The performance of the Pf-21-I assay was compared against Giemsa stained thick blood smears from samples collected at a malaria endemic area of the Bolivar State, Venezuela, at the field station of Malariología in Tumeremo. Coupled to non-radioactive hybridization the pf-21-I performed better than the traditional microscopic method with a r = 1.7:1. In the case of mixed infections the r value of P. falciparum detection increased to 2.5:1. The increased diagnostic sensitivity of the test produced with this homologous oligonucleotide could provide an alternative to the epidemiological diagnosis of P. falciparum being currently used in Venezuela endemic areas, where low parasitemia levels and asymptomatic malaria are frequent. In addition, the DNA fingerprint could be tested in molecular population studies.

Protamine inhibits angiogenesis and growth of C6 rat glioma; a synergistic effect when combined with carmustine.

Protamine inhibits angiogenesis and blocks endothelial, fibroblast and platelet growth factors. Human and experimental gliomas spread and grow in response to both paracrine and autocrine release of these factors. Our objective was to study the effect of protamine administration on cell proliferation, angiogenesis and tumoral growth of C6 glioma. Additionally, we compared the antitumoral effect of protamine with that of another inhibitor of angiogenesis, suramin, and investigated a potential synergistic antitumoral action of low doses of protamine combined with the antineoplastic carmustine. C6 glioma cells were implanted subcutaneously in Wistar rats. A highly malignant glioma developed in 80% of animals; when the tumour reached a diameter of 1.5 cm, either protamine, suramin, carmustine or protamine plus carmustine were administered in various doses. Tumour parameters were measured and compared between groups. In a dose-dependent manner, protamine reduced tumour volume (P < 0.001), mitotic index (P < 0.05), vascular density (P < 0.05) and cell viability (P < 0.005) of C6 glioma. An ultrastructural study demonstrated membranous inclusions in the cytoplasm of 28% of tumoral and endothelial cells of tumours from animals treated with protamine. The inhibition of tumoral growth produced by moderate doses of protamine was similar to that produced by toxic doses of suramin. The combination of protamine and carmustine had a synergistic curtailing effect on tumoral growth (P < 0.001). Our results indicate that protamine is an effective agent against glioblastoma; in non-toxic doses it could potentiate the antineoplastic effect of nitrosoureas for the treatment of glial tumours.

[Fluconazole in meningeal coccidioidomycosis]. / El fluconazol en la coccidioidomicosis meníngea.

We report our experience with ten virgin cases (5 children, 5 adults) with coccidioidal meningitis treated with fluconazole. The diagnosis was confirmed by a positive culture of the cerebrospinal fluid. Fluconazole at doses of 6/mg/kg/day for children and 400 mg/day for adults were given. All responded well, in five it was discontinued and four relapsed but responded well to fluconazole. We conclude that fluconazole is useful for coccidioidal meningitis.