Follow-up of an Asymptomatic Chagas Disease Population of Children after Treatment with Nifurtimox (Lampit) in a Sylvatic Endemic Transmission Area of Colombia

Background

Chagas disease is an anthropozoonosis caused by Trypanosoma cruzi. Two drugs are currently used for the etiological treatment of the disease: Nifurtimox (Lampit) and Benznidazole. This study presents a quasi-experimental trial (non-control group) of sixty-two patients who were treated for Chagas disease with Nifurtimox (Lampit), and were then followed for 30 months post-treatment. The safety of Nifurtimox (Lampit) for Chagas disease in this group of children primarily between 4 and 19 years old was also evaluated.

Materials and methods

The 62 patients included in the study were selected when resulted seropositive for two out of three fundamentally different serological tests. All children were treated during two months according to protocols established by WHO. Monitoring was performed every twenty days to evaluate treatment safety. In 43 patients, two different serological tests: ELISA and IFAT; and two parasitological tests: blood culture, and real time PCR, (qPCR) were performed to assess therapeutic response, defined as post-treatment serological negativization.

Principal findings

All patients completed the treatment successfully, and six patients abandoned the post-treatment follow-up. Adverse effects occurred in 74% of patients, but only 4.8% of cases required temporary suspension to achieve 100% adherence to the 60-day treatment, and all symptoms reverted after treatment completion. Both parasite load (measured through qPCR) and antibodies (ELISA absorbance) evidenced a significant median reduction 6 months after treatment from 6.2 to 0.2 parasite equivalents/mL, and from 0.6 to 0.2 absorbance units respectively (p<0.001). Serological negativization by ELISA was evident since 6 months post-treatment, whereas by IFAT only after 18 months. Serological negativization by the two tests (ELISA and IFAT) was 41.9% (95%CI: 26.5–57.3) after 30 months post-treatment. qPCR was positive in 88.3% of patients pre-treatment and only in 12.1% of patients after 30 months. Survival analysis indicated that only 26.3% (95%CI: 15.5–44.8) persisted with negative qPCR during the whole follow-up period.

Conclusions

Nifurtimox was very well tolerated and successfully reduced parasite load and antibody titers. Re-infection, lysed parasites or a lack of anti-parasitic activity could explain these persistently positive qPCR cases.     

A Randomized Blinded Study of the Left Ventricular Myocardial Performance Index Comparing Epinephrine to Levosimendan following Cardiopulmonary Bypass

Background

The objective was to evaluate the effect of epinephrine and levosimendan on the left ventricle myocardial performance index in patients undergoing on-pump coronary artery by-pass grafting (CABG).

Methods

In a double-blind, randomized clinical trial, 81 patients (age: 45–65 years) of both genders were randomly divided to receive either epinephrine at a dosage of 0.06 mcg.kg¹.min^-1 (epinephrine group, 39 patients) or levosimendan at 0.2 mcg.kg¹.min^-1 (levosimendan group, 42 patients) during the rewarming of cardiopulmonary by-pass (CPB). Hemodynamic data were collected 30 minutes after tracheal intubation, before chest open (pre-CPB) and 10 minutes after termination of protamine (post-CPB). As the primary outcome, we evaluated the left ventricle myocardial performance index by the Doppler echocardiography. The myocardial performance index is the sum of the isovolumetric contraction time and the isovolumetric relaxation time, divided by the ejection time. Secondary outcomes were systolic and diastolic evaluations of the left ventricle and postoperative troponin I and MB-CK levels.

Results

Of the 81 patients allocated to the research, we excluded 2 patients in the epinephrine group and 6 patients in the levosimendan group because they didn’t wean from CPB in the first attempt. There was no statistical difference between the groups in terms of patient characteristics, risk factors, or CPB time. The epinephrine group had a lower left ventricle myocardial performance index (p = 0.0013), higher cardiac index (p = 0.03), lower systemic vascular resistance index (p = 0.01), and higher heart rate (p = 0.04) than the levosimendan group at the post-CPB period. There were no differences between the groups in diastolic dysfunction. The epinephrine group showed higher incidence of weaning from CPB in the first attempt (95% vs 85%, p = 0.0001) when compared to the levosimendan group and the norepinephrine requirement was higher in the levosimenandan group than epinephrine group (16% vs. 47%; p = 0.005) in post-CPB period. Twenty-four hours after surgery, the plasma levels of troponin I (epinephrine group: 4.5 ± 5.7 vs. levosimendan group: 2.5 ± 3.2 g/dl; p = 0.09) and MB-CK (epinephrine group: 50.7 ± 31 vs. levosimendan group: 37 ± 17.6 g/dl; p = 0.08) were not significantly different between the two groups.

Conclusion

When compared to levosimendan, patients treated with epinephrine had a lower left ventricle myocardial performance index in the immediate post-CPB period, encouraging an efficient weaning from CPB in patients undergoing on-pump CABG.

Trial Registration

ClinicalTrials.gov {"type":"clinical-trial","attrs":{"text":"NCT01616069","term_id":"NCT01616069"}}NCT01616069     

Shared Genetic Factors of Anxiety and Depression Symptoms in a Brazilian Family-Based Cohort,the Baependi Heart Study

To investigate the phenotypic and genetic overlap between anxiety and depression symptoms in an admixed population from extended family pedigrees. Participants (n = 1,375) were recruited from a cohort of 93 families (mean age±SD 42±16.3, 57% female) in the rural town of Baependi, Brazil. The Hospital Anxiety and Depression Scale (HADS) was used to assess depression and anxiety symptoms. Heritability estimates were obtained by an adjusted variance component model. Bivariate analyses were performed to obtain the partition of the covariance of anxiety and depression into genetic and environmental components, and to calculate the genetic contribution modulating both sets of symptoms. Anxiety and depression scores were 7.49±4.01 and 5.70±3.82, respectively. Mean scores were affected by age and were significantly higher in women. Heritability for depression and anxiety, corrected for age and sex, were 0.30 and 0.32, respectively. Significant genetic correlations (ρ_g = 0.81) were found between anxiety and depression scores; thus, nearly 66% of the total genetic variance in one set of symptoms was shared with the other set. Our results provided strong evidence for a genetic overlap between anxiety and depression symptoms, which has relevance for our understanding of the biological basis of these constructs and could be exploited in genome-wide association studies.     

Variations in Helicobacter pylori cytotoxin-associated genes and their influence in progression to gastric cancer: implications for prevention

Helicobacter pylori (HP) is a bacterium that colonizes the human stomach and can establish a long-term infection of the gastric mucosa. Persistent Hp infection often induces gastritis and is associated with the development of peptic ulcer disease, atrophic gastritis, and gastric adenocarcinoma. Virulent HP isolates harbor the cag (cytotoxin-associated genes) pathogenicity island (cagPAI), a 40 kb stretch of DNA that encodes components of a type IV secretion system (T4SS). This T4SS forms a pilus for the injection of virulence factors into host target cells, such as the CagA oncoprotein. We analyzed the genetic variability in cagA and other selected genes of the HP cagPAI (cagC, cagE, cagL, cagT, cagV and cag Gamma) using DNA extracted from frozen gastric biopsies or from clinical isolates. Study subjects were 95 cagA+ patients that were histologically diagnosed with chronic gastritis or gastric cancer in Venezuela and Mexico, areas with high prevalence of Hp infection. Sequencing reactions were carried out by both Sanger and next-generation pyrosequencing (454 Roche) methods. We found a total of 381 variants with unambiguous calls observed in at least 10% of the originally tested samples and reference strains. We compared the frequencies of these genetic variants between gastric cancer and chronic gastritis cases. Twenty-six SNPs (11 non-synonymous and 14 synonymous) showed statistically significant differences (P<0.05), and two SNPs, in position 1039 and 1041 of cagE, showed a highly significant association with cancer (p-value = 2.07×10⁻⁶), and the variant codon was located in the VirB3 homology domain of Agrobacterium. The results of this study may provide preliminary information to target antibiotic treatment to high-risk individuals, if effects of these variants are confirmed in further investigations.     

Simulation of quantitative characters by genes with biochemically definable action. III. The components of genetic effects in the inheritance of anthocyanins in Matthiola incana R. Br.

Summary In a self-pollinated plant species, Matthiola incana R. Br., six groups of isogenic lines were developed which were ideally suited for investigating the properties of individual genes controlling a quantitative character. Each group consisted of four homozygous parents for two alleles at each of the two loci in a common genetic background. A complete 4 × 4 diallel cross was obtained in each group. Because of the identical genetic background each diallel set could be considered as a genetic system of two loci. The biochemical functions of the alleles at each locus modifying the structure of the anthocyanin molecule were known. The phenotypes of the nine possible genotypes were qualitatively distinguishable by their flower colour differences. A quantitative measure of the phenotypic value associated with a genotype is the concentration of anthocyanins in flower tissues. In these simplified genetic systems, the nine phenotypic values could be expressed in terms of nine biometrical quantities, eight of which are attributable to the genetic effects of the alleles at the two loci under consideration. An unique solution of the set of nine equations in nine unknowns provided direct estimates of the parameters specifying additive, dominance and epistatic effects. Thus the effects of individual genes in a well-defined genetic background could be estimated by the use of a simple additive genetic model. An extension of the model provided estimates of the genetic parameters in different years and genetic backgrounds.Dominance was found to be the most important type of gene action in the inheritance of anthocyanin content in the flower tissues of M. incana. There was considerable epistasis, but the effect was very unstable over years and genetic backgrounds. The relative magnitude of additive effect was most stable. Heterosis was observed and was found to be largely due to dominance and additive × dominance interactions.

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Zusammenfassung In der selbstbefruchtenden Gartenpflanze Matthiola incana wurde eine Reihe weitgehend isogener Linien entwickelt, die sich in idealer Weise zur Untersuchung der Beiträge einzelner definierter Gene und Genkombinationen zu einem quantitativen Merkmal eignen. Das Material wurde in 6 Gruppen eingeteilt. Jede Gruppe besteht aus 4 homozygoten Eltern, die sich aus der Variation zweier Loci mit je zwei Allelen vor einem gemeinsamen genetischen Hintergrund ableiten und nach dialleler Kreuzung alle kombinatorisch möglichen Genotypen ergeben. Jeder diallele Satz stellt somit ein genetisches System mit zwei Loci dar, dessen qualitative Beiträge zur Modifikation der Anthocyanmoleküle bekannt sind und dessen quantitative Beiträge zur Gesamtkonzentration der Anthocyane gemessen werden. In diesen vereinfachten genetischen Systemen können die Phänotypenwerte der 9 Genotypen durch 9 biometrische Quantitäten beschrieben werden, von denen 8 den genetischen Effekten der variierten Loci und einer dem Beitrag des genetischen Hintergrundes zugeschrieben werden könen. Anhand eines einfachen linearen Modells werden Parameter für die additiven, Dominanz- und Epistasieeffekte der einzelnen Gene aus einem Satz von 9 Gleichungen mit 9 Unbekannten direkt geschätzt. Eine Erweiterung des Modells erlaubt die Schätzung der Parameter in verschiedenen Jahren und mit wechselndem genetischem Hintergrund. Für die Ausbildung des quantitativen Merkmals Anthocyangehalt der Blüten von Matthiola incana erwiesen sich die Dominanzkomponenten als der entscheidende Typ der Genwirkung. Ferner wurde ein erheblicher Epistasieanteil geschätzt, jedoch waren diese Beiträge nicht konstant, da sie über die Jahre variierten und eine starke Abhängigkeit von der Art des genetischen Hintergrundes zeigten. Der relative Anteil der additiven Komponenten erwies sich dagegen zwar als gering, jedoch sehr stabil. Beobachtete Heterosiseffekte müssen zum Teil den Dominanzkomponenten, zum Teil der Interaktion zwischen additiven und Dominanzkomponenten zugeschrieben werden.

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This work was carried out while the senior author held a research fellowship awarded by the Alexander von Humboldt-Stiftung of Bad Godesberg, Germany, which is gratefully acknowledged.

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Research supported by Deutsche Forschungsgemeinschaft.     

Simulation of quantitative characters by genes with biochemically definable action

Summary Six groups of genetic materials were developed in a cruciferous garden plant, Matthiola incana R. Br., to produce simplified genetic systems of a pair of loci in each group. There were only two alleles at each locus, which were directly involved in the modification of anthocyanins in the plant tissue. The parental lines and their F₁'s in each group constituted an ideal 4 × 4 diallel cross and satisfied all but one condition necessary for a valid diallel analysis. Nonallelic interaction was the only possible offending postulate.The diallel analysis of the data on anthocyanin content in the flower tissues and a comparison of the results with that of a relatively straightforward method of analysis indicated that in the presence of epistasis, the dominance ratio (H₁/D) ceases to be a reliable measure of the average degree of dominance. In such situations additional genetic information obtained from the diallel analysis are not in agreement with the expectations on the basis of the already available genetic information on the materials. The estimator H₂/4 H₁, a measure of average value of the product of alleles with positive and negative effects, seemed to remain unaffected by epistasis. The Wr/Vr regression analysis does not always permit the detection of nonallelic gene interactions. The results suggest that duplicate interaction may escape detection by the regression analysis.

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Zusammenfassung Anhand der Crucifere Matthiola incana wurde ein genetisches Material aufgebaut, das es erlaubt, in 6 verschiedenen Gruppen je 2 Loci in allen Kombinationen systematisch zu variieren. Je Locus liegen 2 Allele vor, die in die Modifikation der Anthocyane eingreifen.Die Elterlinien und ihre F₁-Nachkommen bilden ein ideales 4 × 4-Diallel und erfüllen bis auf eine alle für eine gültige Diallelanalyse erforderlichen Bedingungen. Die einzige nicht erfüllte Voraussetzung ist die Abwesenheit nichtalleler Interaktionen.Die diallele Analyse der Daten über den Anthocyangehalt der Petalen und ein Vergleich der Resultate mit den aufgrund einer sehr direkten und detaillierten Untersuchungsmethode erhaltenen zeigten, daß das Dominanzverhältnis (H₁/D) bei Vorliegen von Epistasie keine brauchbare Messung des durchschnittlichen Dominanzgrades mehr ist. In derartigen Situationen stimmen auch die zusätzlich aus der diallelen Analyse zu erhaltenden Informationen nicht mit den Erwartungen überein, die auf der bereits vorliegenden Kenntnis der Genetik des Materials beruhen.Der Schätzwert H₂/4 H₁ als Messung des Durchschnittswertes des Produktes der Allele mit positiven und negativen Effekten scheint dagegen durch Epistasie unbeeinflußt zu bleiben.Die W _r /V _r , Regressionsanalyse führt nicht in allen Fällen zum Nachweis nichtalleler Interaktionen. Die Ergebnisse zeigen, daß eine duplicate gene action dem Nachweis durch die Regressionsanalyse entgeht.

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This work was carried out while the senior author held a research fellowship awarded by the Alexander von Humboldt-Stiftung, Bad Godesberg, Germany, which is gratefully acknowledged.Research supported by Deutsche Forschungsgemeinschaft.     

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