Adela Sembaj

Risk progression to chronic Chagas cardiomyopathy: influence of male sex and of parasitaemia detected by polymerase chain reaction

Background: Polymerase chain reaction (PCR) allows detection of Trypanosoma cruzi in blood throughout the course of Chagas’ disease. Objective: To determine whether T cruzi DNA detected by PCR is associated with progression to chronic Chagas cardiomyopathy. Design: Prospective cohort study. Setting: A tertiary care centre in Argentina. Patients: 56 consecutive patients with chronic T cruzi infection. Methods: Clinical examination, ECG, and Doppler echocardiography were carried out at baseline and at the end of the follow up. Detection of T cruzi DNA by PCR amplifying a nuclear sequence was undertaken in all patients at baseline. Main outcome measures: Progression was defined as death from chronic cardiomyopathy or the presence of a new ECG or left ventricular echocardiographic abnormality at the end of follow up. Results: The 56 patients (21 male, 35 female; mean (SD) age, 56.0 (11.3) years) were followed for a mean 936.3 (244.39) days. Progression to cardiomyopathy was detected in 12 patients (21.4%). Three of these patients died after baseline evaluation. Univariate analysis showed that a positive PCR (relative risk 4.09, 95% confidence interval (CI) 1.60 to 9.85) and male sex (5.00, 95% CI 1.65 to 15.73) were associated with progression. Multivariable logistic regression indicated that both sex and PCR were independent variables affecting the outcome. Conclusions: In a cohort of seropositive individuals, patients with T cruzi DNA detected by PCR and male patients were at higher risk of progression. These results highlight the importance of T cruzi in the pathophysiology of chronic cardiomyopathy.

Trypanosoma cruzi in Persons without Serologic Evidence of Disease, Argentina

Current diagnosis of chronic Chagas disease relies on serologic detection of specific immunoglobulin G against Trypanosoma cruzi. However, the presence of parasites detected by polymerase chain reaction (PCR) in patients without positive conventional serologic testing has been observed. We determined the prevalence and clinical characteristics of persons with seronegative results for T. cruzi DNA detected by PCR in a population at high risk for chronic American trypanosomiasis. We studied a total of 194 persons from two different populations: 110 patients were recruited from an urban cardiology clinic, and 84 persons were nonselected citizens from a highly disease-endemic area. Eighty (41%) of persons had negative serologic findings; 12 (15%) had a positive PCR. Three patients with negative serologic findings and positive PCR results had clinical signs and symptoms that suggested Chagas cardiomyopathy. This finding challenges the current recommendations for Chagas disease diagnosis, therapy, and blood transfusion policies.

Prevalence of Circulating Trypanosoma cruzi Detected by Polymerase Chain Reaction in Patients With Chagas' Cardiomyopathy

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Polymerase chain reaction procedure to detect Trypanosoma cruzi in blood samples from chronic chagasic patients

The feasibility of DNA amplification by the polymerase chain reaction for specific detection of Trypanosoma cruzi in human blood was investigated. We have used primers flanking a 220-bp fragment of highly repetitive elements, the E13 element, in T. cruzi nuclear DNA. Only polymerase chain reaction products from blood samples of chronic chagasic patients showed several amplified fragments in 1.6% agarose gels stained with ethidium bromide. Southern blot analysis demonstrated that the 220-bp amplified fragment is specific for T. cruzi DNA and very useful to detect the presence of the parasite in blood from chronic chagasic patients.

Periodontal conditions during the pregnancy associated with periodontal pathogens

AIM To describe the bacterial associations in the periodontal pockets of pregnant women and to correlate the presence of Prevotella intermedia, Tannerella forsythia (T. forsythia), Treponema denticola (T. denticola), Aggregatibacter actinomycetemcomitans, and Porphyromona gingivalis (P. gingivalis) with periodontal parameters of severity. METHODS The analysis was performed with 150 pregnant women. The examination consisted of an evaluation of bleeding, suppuration, probing depths, clinical attachment levels, hypermobility scores, the Silness and Löe Plaque Index, and the Löe and the Silness Gingival Index. Each periodontal pathogen was identified by polymerase chain reaction. RESULTS A statistically-significant association was observed (P < 0.01) between P. gingivalis and T. forsythia, between P. gingivalis and T. denticola, and between T. forsythia and T. denticola. Age was observed to be a risk factor in the development of moderate periodontitis (odds ratio [OR] = 4.92, 95% confidence interval [CI] = 1.1-21.3, P = 0.0328). Age was significantly associated with increased pocket depth and plaque index (OR = 6.36, 95% CI = 1.8-22.2, P = 0.0037). In pregnant women, the presence of P. gingivalis was found to increase the risk of developing a clinical attachment level ≥ 5 mm. CONCLUSION A high prevalence of P. gingivalis in pregnant women, especially in combination with T. forsythia and T. denticola, was associated with an increased risk of developing moderate periodontitis, and that association was more marked in pregnant women aged 30 years or older.

Association between maternal periodontitis and preterm and/or low birth weight infants in normal pregnancies

Abstract Objective: To relate the periodontal condition with the presence of periodontal bacteria in pregnant that had babies with preterm delivery or/and low weight at birth (PTLBW). Methods: We recruited 134 pregnant women without systemic diseases attending at the Gynaecology and Obstetrics Room, from Maternal Provincial Hospital, Córdoba, Argentine. Pregnant were grouped according to the International Classification for a System of Periodontal Disease. A sample from periodontal pocket was extracted to identify Porphyromonas gingivalis (Pg), Treponema denticola (Td), Tannerella forsythia (Tf) Prevotella intermedia (Pi) and Agreggatibacter actinomycemcomitans (Aa). Results: We identified 7 (5%) cases of children born underweight or preterm of mothers diagnosed with Gingivitis, 6 (4%) in Mild Periodontitis and 4 (3%) in Moderate Periodontitis. We estimated that when Pi and/or Aa were not detected in the periodontal pockets of mothers, the infants had more than 129% chance of having normal birth weights (OR 3.47 for Pi and OR and 2.29 for Aa). The average age of the mothers who has PTLBW was 21 ± 3.5. The age showed an association with PTLBW (p < 0.0008). Conclusions: The presences of periodontal pathogens in periodontal pockets from pregnant with different periodontal status would associate with PTLBW infants when the mothers are young, and the normal term and normal birth weight infant are associated with the absence of periodonto bacteria like Pi and Aa.

Alkaline phosphatase isoenzymes in plasma of chagasic and healthy pregnant women

Catedra de Quimica Biologica, Facultad de CienciasMedicas **Catedra de Estadistica y Biometria,Facultad de Ciencias Exactas, Fisicas y Naturales,Universidad Nacional de Cordoba, Pabellon Argentina2° Piso, Ciudad Universitaria, Cordoba, Argentina*Servicio Nacional de Chagas, Cordoba, ArgentinaKey words: alkaline phosphatase - pregnancy -Chagas disease

Polimorfismos de un solo nucleótido en genes de endotelina-1 y su receptor A asociados a daño cardiovascular en hipertensión arterial esencial

INTRODUCTION The endothelin system, for its vasoconstrictor action, is related to the development of essential hypertension (HTAe). The polymorphism analysis of their genes represents a new approach to the study of this disease. We propose to analyze the interaction between stages of essential hypertension (HTAe) and risk factors with polymorphisms 138ex1 ins/del A gene endothelin-1 (ET-1) and H323H receptor gene A ET-1 (ETRA). PATIENTS AND METHODS We included 300 patients of both sexes, unrelated, who consecutively attended the clinic hypertension medical service. Each one underwent a complete physical examination, electrocardiogram, echocardiogram, and Rx thorax. The degree of severity of hypertension was determined in stages. The determination of polymorphisms was performed by amplification followed by cutting by specific restriction enzyme from DNA obtained from peripheral blood. RESULTS The 46% of patients had HTAe controlled, 17.6% had organ damage or cardiovascular, brain or kidney disease. It was observed that the 4A/4A carriers showed lower frequency of cardiovascular disease, kidney and brain (P<.032; 95% CI: 11.1-21.4). For H323H polymorphism, the evaluation by images showed a higher frequency of the dilations of left auricular (P=.02) and auricular fibrillation (P=.03) between the T/T carrier, a higher frequency of cardiomegaly was detected in C/C patients (P=.04). CONCLUSION The genotypes, 4A/4A of the ET-1 gene and the T/T from ETRA gene might be involved in worse outcome of cardiovascular damage. Their identification could help recognize subgroups of the hypertensive patients with different risk.

Evaluation of periodontal mechanical therapy in deep pockets: Clinical and bacteriological response

Objectives: To evaluate the microbial composition and clinical parameters of periodontal pockets with probing depth ≥5 mm at baseline, 1 week, 3 and 12 months after scaling and root planning. Methods: Clinical parameters were measured and bacterial samples were collected from 44 sites in 11 patients with chronic periodontitis. By means of Polymerase Chain Reaction (PCR) the presence of Aggregatibacter actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), Treponema denticola (Td), Tannerella forsythia (Tf) and Prevotella intermedia (Pi) was estimated. The patients received mechanical periodontal therapy and were evaluated after 1 week, 3 months and 12 months. Results: After treatment, all clinical parameters (Plaque, Bleeding on Probing, Supuration, Probing Pocket Depth and Clinical Attachment Level) were significantly reduced, and the values obtained were maintained up to the 12 months that the study lasts. At baseline, the most prevalent species were Pg, present in 66% of the sites, Tf (55%) and Td (41%). The deepest sites were related to the association Tf-Td (6.8 mm) and Tf-Td-Pi (7 mm). The number of positive sites for Td, Tf and Pg was significantly reduced after therapy. Conclusions: Scaling and root planning improve significantly clinical parameters as well as reduce the prevalence of periodontal pathogens Pg, Td and Tf in deep periodontal pockets. The results obtained were maintained up to 12 months. No further clinical attachment loss was found in 86% of the sites at 3 months and 79% at 12 months. The sites where the treatment failed in removing pathogens developed at 12 months greater probing pocket depths. Rev. Clin. Periodoncia Implantol. Rehabil. Oral Vol. 5(3); 123-127, 2012.

Determination of alkaline phosphatase isozymes in amniotic fluid

A simple method for the determination of the three isozymes of alkaline phosphatase (EC 3.1.3.1) contained in amniotic fluid (fetal intestinal, placental, and liver-bone-kidney) is presented. Total alkaline phosphatase activity was assayed in 10,000 g supernatants of amniotic fluid from 30 normal women between the 16th and 20th week of pregnancy. Electrophoretic patterns and inhibition by L-phenylalanine and L-homoarginine studies showed that all the fetal intestinal isozyme was precipitated in the pellet after centrifugation at 100,000 g for 90 min. Thus, the difference between total alkaline phosphatase activity and activity in the 100,000 g supernatant corresponds to fetal intestinal alkaline phosphatase. Placental isozyme can be determined by assaying alkaline phosphatase in the 100,000 g supernatant after heating at 56 degrees C for 90 min. Liver-bone-kidney isozyme activity is obtained by subtracting placental alkaline phosphatase activity from that of the 100,000 g supernatant. Mean percentages of the total alkaline phosphatase for each of the isozymes in amniotic fluid were 81% for fetal intestinal alkaline phosphatase, 7.5% for placental alkaline phosphatase and 12.0% for liver-bone-kidney alkaline phosphatase. Determination of fetal intestinal alkaline phosphatase by this method could be applied to the diagnosis of cystic fibrosis in fetuses having a 1:4 risk of being affected.