Schistosoma mansoni: evaluation of an RNAi-based treatment targeting HGPRTase gene.

Hypoxanthine-guanine phosphoribosyltransferase (HGPRTase) is an essential gene of the parasite Schistosoma mansoni and it is well conserved in its hosts (mouse and human) at the protein but not at the RNA level. This feature prompted us to assess RNA interference (RNAi) to combat schistosomiasis. Small interfering RNAs (siRNAs) were produced against HGPRTase, injected in infected mice and the number of worms was counted six days after injection. The total number of parasites was reduced by approximately 27% after treatment. RT-PCR analyzes showed a significant reduction in parasite target mRNA but not in host's homologue. The use of low doses of molecules did not oversaturate si- or miRNA pathways as mice survival rates were not affected by siRNAs. This is the first successful in vivo demonstration of a RNAi-based treatment against schistosomiasis. We believe that improvements in molecule delivery and an increase on siRNA dose could rapidly eliminate parasite.

Molecular characterisation and evolutionary relationships of a potyvirus infecting Crotalaria in Brazil. Brief report.

The 3' terminal genomic region of a potyvirus causing mosaic disease in several Crotalaria species has been cloned and sequenced. Comparisons of the nucleotide and deduced amino acid (aa) sequences of the cloned cDNA with those from other potyviruses show that the Crotalaria-infecting virus (designated Crotalaria mosaic virus; CrMV) is closely related to Cowpea aphid-borne mosaic virus (CABMV). Maximum identity (95.4%) at the coat protein (CP) aa level was observed between CrMV and a Brazilian strain of CABMV. Phylogenetic analyses derived from the sequence alignments of the CP and 3' untranslated region confirmed the identification of CrMV as a strain of CABMV and the name CABMV-Cr is suggested.

Mitochondrial uncoupling proteins in mammals and plants.

Uncoupling proteins (UCPs) belong to a distinct cluster of the mitochondrial anion carrier family. Up to five different uncoupling protein types were found in mitochondria of mammals and plants, and recently in fishes, fungi and protozoa. They exhibit a significantly conserved structure with several motifs specific to either the whole cluster or protein type. Uncoupling proteins, as well as the whole mitochondrial anion carrier gene family, probably emerged in evolution before the separation of animal, fungi, and plant kingdoms and originate from an anion/nucleotide or anion/anion transporter ancestor. Mammalian UCP1, UCP2, UCP3, and plant uncoupling proteins pUCP1 and pUCP2 are similar and seem to form one subgroup, whereas UCP4 and BMCP1 belong to a different group. Molecular, biochemical, and phylogenic data suggest that UCP2 could be considered as an UCP-prototype. UCP1 plays its biological role mainly in the non-shivering thermogenesis while the role of the other types is unknown. However, hypotheses have suggested that they are involved in the general balance of basic energy expenditure, protection from reactive oxygen species, and, in plants, in fruit ripening and seed ontogeny.

Functional reconstitution of Arabidopsis thaliana plant uncoupling mitochondrial protein (AtPUMP1) expressed in Escherichia coli.

The Arabidopsis thaliana uncoupling protein (UCP) gene was expressed in Escherichia coli and isolated protein reconstituted into liposomes. Linoleic acid-induced H+ fluxes were sensitive to purine nucleotide inhibition with an apparent K(i) (in mM) of 0.8 (GDP), 0.85 (ATP), 0.98 (GTP), and 1.41 (ADP); the inhibition was pH-dependent. Kinetics of AtPUMP1-mediated H+ fluxes were determined for lauric, myristic, palmitic, oleic, linoleic, and linolenic acids. Properties of recombinant AtPUMP1 indicate that it represents a plant counterpart of animal UCP2 or UCP3. This work brings the functional and genetic approaches together for the first time, providing strong support that AtPUMP1 is truly an UCP.

Arrhythmogenic cardiomyopathy of the right ventricle. Predictive value of QT interval dispersion to assess arrhythmogenic risk and sudden death.

OBJECTIVE: To determine in arrhythmogenic right ventricular cardiomyopathy the value of QT interval dispersion for identifying the induction of sustained ventricular tachycardia in the electrophysiological study or the risk of sudden cardiac death. METHODS: We assessed QT interval dispersion in the 12-lead electrocardiogram of 26 patients with arrhythmogenic right ventricular cardiomyopathy. We analyzed its association with sustained ventricular tachycardia and sudden cardiac death, and in 16 controls similar in age and sex. RESULTS: (mean +/- SD). QT interval dispersion: patients = 53.8+/-14.1 ms; control group = 35.0+/-10.6 ms, p = 0.001. Patients with induction of ventricular tachycardia: 52.5+/-13.8 ms; without induction of ventricular tachycardia: 57.5+/-12.8 ms, p =0.420. In a mean follow-up period of 41+/-11 months, five sudden cardiac deaths occurred. QT interval dispersion in this group was 62.0+/-17.8, and in the others it was 51.9+/-12.8 ms, p = 0.852. Using a cutoff > or =60 ms to define an increase in the degree of the QT interval dispersion, we were able to identify patients at risk of sudden cardiac death with a sensitivity of 60%, a specificity of 57%, and positive and negative predictive values of 25% and 85%, respectively. CONCLUSION: Patients with arrhythmogenic right ventricular cardiomyopathy have a significant increase in the degree of QT interval dispersion when compared with the healthy population. However it, did not identify patients with induction of ventricular tachycardia in the electrophysiological study, showing a very low predictive value for defining the risk of sudden cardiac death in the population studied.

The Brugada syndrome. Outcome of one case.

The Brugada syndrome is a rare condition, and due to its mutating manner of presentation it may be difficult to diagnose. We report one case and discuss the diagnostic aspects and the clinical outcome of one patient with characteristic findings of this syndrome. These findings are especially defined by J-ST elevation in the right leads of serial electrocardiographic records, wide oscillations of J points and ST segments during 24-hour Holter monitoring, and nocturnal sudden death. We stress the importance of the Holter monitor findings for diagnostic complementation. Through this method it is possible to establish a correlation between vigil activities and sleep and the variability of the degree of impairment in ventricular repolarization.

Electrical behavior of T-wave polarity alternans in patients with congenital long QT syndrome.

OBJECTIVES: This study was designed to evaluate the incidence and characteristics of onset of T-wave polarity alternans (TWPA) in patients with long QT syndrome. BACKGROUND: The T-wave alternans is a phenomenon that consists of beat-to-beat variability in the amplitude, morphology, and sometimes polarity of the T-wave, and it may trigger life-threatening arrhythmias. METHODS: The 24-h Holter recordings of 11 patients with congenital long QT syndrome were studied. Episodes of TWPA with 10 or more consecutive cycles were selected and analyzed as follows: 1) mean cycle length (MCL) and QTc interval duration (QTcI) of the episodes of TWPA and the 10 cycles preceding and succeeding the TWPA; 2) MCL and QTcI of the third, second, and first minute before onset (Mn_3, Mn_2, Mn_1); 3) MCL and QTcI from the tenth to the first cycle immediately preceding the onset of TWPA (R_10 to R_1); 4) MCL and QTcI from the first to the fourteenth cycle during alternans (R0 to R14); 5) MCL and QTcI from the first to the tenth cycle immediately succeeding TWPA (R+1 to R+10); 6) linear correlation (Lnc) between QT interval and cycle length (CL) (LncQT/CL) during alternans and for the 10 preceding cycles; 7) Lnc between the first three alternans cycles and episode duration (Lnc 3CL/EpD); and 8) difference between the longest and shortest QTc interval. We also selected episodes consisting of four or more consecutive cycles in order to analyze daily rhythms of the phenomenon. RESULTS: The TWPA was observed in 5 (45%) out of the 11 patients studied. The alternans process is initiated by a sudden shortening of the first alternans cycle without previous heart rate changes and ends at the moment when prolongation of the cycle tends to occur. LncQT/ CL-alternans: r = 0.38 +/- 0.2 (p = 0.20); without alternans: r = 0.81 +/- 0.06 (p = 0.01). Lnc 3CL/EpD: r = 0.002 (p = 0.992). The QTc difference during alternans: 312.0 +/- 52.1 ms; without alternans: 86.0 +/- 36.4 ms (p = 0.001). Daily rhythm: 71% of the episodes occurred between 8 AM and 8 PM, with higher incidence during the morning. CONCLUSIONS: The TWPA was dependent on the cardiac CL; there was loss of the LncQT/CL and an increase in the QT interval variability. Like other biological variables, T-wave polarity alternans has a higher density during the morning.

Molecular evidence that sugarcane yellow leaf virus (ScYLV) is a member of the Luteoviridae family.

A previously uncharacterized virus was reported in southeast Brazil causing a yellowing leaf disease in sugarcane. The virus, termed sugarcane yellow leaf virus (ScYLV), shares features typical of the luteoviruses. To start the molecular characterization of ScYLV, the nucleotide sequence of the coat protein (CP), 17 kDa protein and C-terminus of the RNA-dependent RNA polymerase coding regions was determined from an RT-PCR amplification product. Comparisons showed that the deduced amino acid sequences share a considerable degree of identity and similarity with corresponding sequences of known luteoviruses, thus clearly establishing ScYLV as a member of the family Luteoviridae. The authenticity of the CP open reading frame was confirmed by its expression in Escherichia coli. The recombinant CP positively reacted in immunoblot assays with polyclonal antibodies raised against native ScYLV. Furthermore, phylogenetic analyses also suggest that the 5' and 3' coding blocks of the ScYLV genome possess different taxonomic affinities within the Luteoviridae family, as does also the genome of soybean dwarf virus.

Deletion mapping of the potyviral helper component-proteinase reveals two regions involved in RNA binding.

The Potyvirus helper component-proteinase (HC-Pro) binds nonspecifically to single-stranded nucleic acids with a preference for RNA. To delineate the regions of the protein responsible for RNA binding, deletions were introduced into the full-length Potato potyvirus Y HC-Pro gene carried by an Escherichia coli expression vector. The corresponding proteins were expressed as fusions with the maltose-binding protein, purified, and assayed for their RNA-binding capacity. The results obtained by UV cross-linking and Northwestern blot assays demonstrated that the N- and C-terminal regions of HC-Pro are dispensable for RNA binding. They also revealed the presence of two independent RNA-binding domains (designated A and B) located in the central part of HC-Pro. Domain B appears to contain a ribonucleoprotein (RNP) motif typical of a large family of RNA-binding proteins involved in several cellular processes. The possibility that domain B consists of an RNP domain is discussed and suggests that HC-Pro could constitute the first example of a plant viral protein belonging to the RNP-containing family of proteins.

Low temperature and aging-promoted expression of PUMP in potato tuber mitochondria.

In this communication, we show that the plant uncoupling mitochondrial protein (PUMP) present in potato tuber mitochondria is induced by aging at 28 degrees C and that this induction is strongly stimulated when the potato tubers are stored at low temperature (4 degrees C). PUMP activity was detected by the degree of linoleic acid (LA)-induced ATP-sensitive mitochondrial uncoupling measured as a function of the decrease in membrane potential (delta psi). The PUMP content was evaluated by immunoblot analysis using polyclonal antibodies raised against potato PUMP that specifically detected a 32 kDa band. In agreement with the effect of LA on delta psi, the content of the 32 kDa band increased during storage and was stimulated by low temperature. These results support the proposed role of PUMP in plant thermogenesis and possibly in fruit ripening and senescence.

[Contribution of dynamic electrocardiography by Holter monitoring in the evaluation of congenital long QT syndrome patients]. / Contribuição de eletrocardiografia dinâmica pelo sistema Holter na avaliação de pacientes com a síndrome congênita do QT longo.

PURPOSE: The purpose of this study was to evaluate the value of ambulatory electrocardiogram as a clinical tool to assess ventricular repolarization in patients with the congenital long QT syndrome. METHODS: The study population comprised six patients and their data were compared to a control group of six patients matched in age and gender. The QT interval (ms), corrected by the heart rate, was measured in the first minute of each hour using two monitoring leads, with the mean of six consecutive complexes. The data obtained include the morphologic pattern of T wave, the mean 24-h QTc interval, relation between QT and cardiac cycle, QTc variability (assessed calculating hourly standard deviation of the interval and then obtaining the global 24-h mean), QTc dispersion (difference between the longest and shortest QTc interval). RESULTS: In all patients abnormal patterns of T waves were detected with frequent episodes of T wave alternans. Mean 24-h QTc--patients: 598.2 +/- 73.8 ms; controls: 436.1 +/- 8.9 ms (p = 0.000). Linear correlation and regression between QT and heart rate-patients: r = 0.812; controls: r = 0.967 (p = 0.000). QTc variability-patients: 36.9 +/- 17.2 ms; controls: 14.7 +/- 2.1 ms (p = 0.01). QTc dispersion-patients: 168.3 +/- 70.2 ms; controls: 53.3 +/- 8.1 ms (p = 0.000). CONCLUSION: The data showed increased hourly QTc variability. QTc dispersion and worse correlation between QT and heart rate. This data may reflect an abnormally augmented ventricular vulnerability.

[Evaluation of the ventricular arrhythmogenic profile of patients with dilated cardiomyopathy and reduced ejection fraction]. / Avaliação do perfil arritmogênico ventricular de pacientes com miocradiopatia dilatada e fralcão de Ejeção diminuída.

PURPOSE: To evaluate the arrhythmogenic profile of patients with dilated cardiomyopathy of low ejection fraction and its prognostic significance. METHODS: Data from 40 patients (30 males; mean age: 52 +/- 13 years) were analysed including ventricular arrhythmias (24h - Holter monitoring), autonomic balance from heart rate variability in time domain (rMSSD and pNN50 indexes), ventricular late potentials (signal averaged electrocardiogram (ECG) and dispersion of ventricular repolarization measured from 12-lead ECG. RESULTS: There was a high prevalence of ventricular arrhythmias with at least one episode of nonsustained ventricular tachycardia (VT) in 60% of the patients. Depressed vagal activity was observed in more than half of the patients. In only 30% of the patients the signal-averaged ECG was positive. The dispersion of ventricular repolarization ranged from 20 to 100 ms. The presence of > 30 ventricular premature beats or nonsustained VT on Holter monitoring was the most significant predictor of cardiac death and sudden cardiac death with a relative risk of 1.9 and 3.2, respectively (p = 0.01 and 0.000). CONCLUSION: In this study population it was noted that patients with dilated cardiomyopathy and low ejection fraction had an abnormal electrical and autonomic cardiac behaviour. These findings could represent risk factors for the occurrence of life-threatening arrhythmias or fatal events.

[Sinus node modification by catheter using radiofrequency current in a patient with inappropriate sinus tachycardia. Evaluation of early and late results]. / Modificação do nódulo sinusal via cateter por energia de radiofreqüência em paciente com taquicardia sinusal inapropriada. Avaliação dos resultados imediatos e tardios.

This article reports the early and late results of a patient in whom radiofrequency current was used to ablate an incessant inappropriate sinus tachycardia. During successful radiofrequency application there was a sudden change of rate and subsequent emergence of a stable rhythm with the same sinus node P wave characteristics. During follow-up, normal cardiac cycles were still present after six months of the procedure, although the patient still complained of palpitations, suggesting no correlation with the heart rate.

AtPUMP: an Arabidopsis gene encoding a plant uncoupling mitochondrial protein.

A cDNA clone (AtPUMP) encoding a plant uncoupling mitochondrial protein was isolated from Arabidopsis thaliana. The cDNA contains an open reading frame of 921 nucleotides encoding 306 amino acids (predicted molecular weight 32,708). The predicted polypeptide is 81% identical and 89% similar to the potato UCP-like protein, and includes an energy transfer protein motif common to mitochondrial transporters. The AtPUMP gene exists as a single copy in the Arabidopsis genome. The corresponding transcript was expressed in all tissues and was strongly induced by cold treatment. We suggest that the putative AtPUMP protein may play a role in heat-requiring physiological events in Arabidopsis.

Contribuição de eltrocardiografia dinâmica pelo sistema holter na avaliação de pacientes com a síndrome congênita do QT longo / Contribution of the dynamic electrocardiography by holter monitoring in the evaluation of congenital long QT syndrome patients

OBJETIVO - Avaliar pelo Holter-24h a dinâmica da repolarização ventricular de pacientes com a síndrome congênita do QT longo. MÉTODOS - Foram incluídos seis pacientes, sendo os resultados confrontados com os observados em um grupo controle semelhante em número, idade e sexo. Analisaram-se nas gravaçöes a morfologia da onda T e sua dinâmica, o intervalo QTc médio para as 24h, as relaçöes entre QT e ciclo cardíaco, a variabilidade do QTc das 24h, bem como a sua dispersão (diferença entre o maior e menor QTc observado). Todas as variáveis foram definidas a partir da medida do intervalo QT obtida no 1§ minuto de cada hora da gravação. RESULTADOS - Alteraçöes morfológicas da onda T estiveram presentes em todos os pacientes, observando-se em cindo deles freqüentes episódios de alternância da onda T, achados ausentes no controle. QTc médio para as 24h - controle: 436,1ñ8,9ms; pacientes: 598,2ñ73,8ms (p=0,000). Correlação linear entre QT e ciclo cardíaco - controle: r=0,967; pacientes: r=0,812 (p=0,000). Variabilidade do QTc para as 24h - controle: 14,7ñ2,1ms; pacientes: 36,9ñ17,1ms (p=0,01). Dispersão do QTc - controle: 53,3ñ8,1ms; pacientes: 168,3ñ70,2ms (p=0,000). CONCLUSÄO - Os resultados apresentaram ampla variabiblidade dos intervalos QTc nas 24h, menor adaptabiblidade do intervalo QT às flutuaçöes dos ciclos cardíacos e significativa dispersão da repolarização ventricular nas 24h, confirmando mais uma vez a existência de importante alteração da vulnerabilidade ventricular nesses pacientes.

Dispersäo temporal da repolarizaçäo ventricular, alternância elétrica da onda T e variabilidade dos intervalos Qt: metodologia de aquisiçäo, análise dos dados, importância clínica / Temporal dispersion of the ventricular repolarization, T wave eletrical alternans and Q-T intervals variability. Methodology analysis of data and clinical importance

No presente artigo, os autores fazem uma revisäo da metodologia e da importância clínica de alguns novos métodos de análise da repolarizaçäo ventricular pelo eletrocardiograma de superfície ou utilizando a eletrocardiograma de superfície ou utilizando a eletrocardiografia dinâmica pelo sistema Holter. Incluem,no estudo, a análise da dispersäo temporal da repolarizaçäo ventricular, a variabilidade dos intervalos Qt e a alternância elétrica da onda T. Acentuam a importância e as limitaçöes desses métodos na estratificaçäo de risco arritmogênico em algumas cardiopatias e as perspectivas dos mesmos com os estudos em andamento que tentam desenvolver programas e algoritmos que permitam uma correta determinaçäo automática das variáveis envolvidas

Fibrilação e flutter atriais / Atrial fibrillation and flutter

No presente artigo os autores fazem uma breve revisão da clínica dos mecanismos eletrofisiológicos do Flutter e Fibrilação atriais. Procuram de forma sucinta fazer uma atualização terapêutica, uma vez que o grande avanço ocorrido na última década na área de eletrofisiologia clínica possibilitou não apenas um melhor conhecimento dos mecanismos das arritmias cardíacas e o mecanismo de ação das drogas antiarrítmicas, como também o tratamento não farmacológico das mesmas. Nesta revisão, os autores procuram abranger o tratamento farmacológico das arritmias, a indicação atual para a anticoagulação na prevenção dos fenômenos tromboembólicos e avaliar as indicaçöes atuais, sucessos e limitaçöes da terapia não farmacológica dos distúrbios do ritmo em questão.

Avaliaçäo do perfil arritmogênico ventricular de pacientes com miocardiopatia dilatada e fraçäo de ejeçäo diminuída / Evaluation of the arrhythmogenic profile of patients with dilated cardiomyopathy and reduced ejection fraction

OBJETIVO - Avaliar o perfil arritmogênico ventricular de pacientes com miocardiopatia dilatada e fraçäo de ejeçäo diminuída. MÉTODOS - Estudo prospectivo em 40 pacientes com fraçäo de ejeçäo média ao ecocardiograma de 32,5ñ2,1 'por cento', obtida pela análise do Holter de 24h, do balanço autonômico cardíaco, determinado por índices de variabilidade da freqüência cardíaca (rMSSD e pNN50), do eletrocardiograma (ECG) de alta resoluçäo (ECGAR) e do grau de dispersäo da repolarizaçäo ventricular determinado no ECG de superfície. Por regressäo logística determinaram-se, a partir dos resultados, os preditores de risco para morte cardíaca e morte súbita. RESULTADOS - Observou-se na populaçäo envolvida uma elevada incidência de ectopias ventriculares isoladas, pareadas e de surtos näo sustentados de taquicardia ventricular. Pela análise do balanço autonômico notou-se depressäo da atividade vagal cardíaca em mais da metade dos pacientes, sendo que apenas 30 'por cento' apresentaram ECGAR positivo. O grau de dispersäo temporal da repolarizaçäo ventricular variou de 20 a 100ms. A presença de > 30 extra-sístoles isoladas por hora e de taquicardia ventricular näo sustentada ao Holter foram os preditores de risco com valores mais elevados para a morte cardíaca e morte súbita, com uma razäo de risco respectiva de 1,9 e 3,2 (p=0,01 e 0,000). CONCLUSÄO - Foram observadas importantes alteraçöes no comportamento elétrico e autonômico cardíaco, constituindo-se fatores de risco para a ocorrência de eventos arrítmicos graves ou fatais.

[Relationship between mitral valve prolapse and arrhythmogenic right ventricular disease]. / Associação entre prolapso valvar mitral e doença ventricular direita arritmogênica.

PURPOSE: To evaluate some features of ventricular arrhythmias in patients with mitral valve prolapse. METHODS: We studied 25 patients (female: 19; mean age: 37 +/- 13 years) with ventricular arrhythmias, mitral valve prolapse and normal ventricular function. All patients underwent a 24h Holter and high resolution ECG (HRECG). The Qtc intervals were measured in lead II (normal < 0.44 s). In order to define the possible origin of the ventricular focus, the morphology of the ectopic beats were analysed in leads I, II, aVF, V1 using the following criteria: 1) LBBB morphology with left axis deviation in the frontal plane (FP): origin at the inflow tract of the right ventricle (RV); 2) LBBB morphology with right axis deviation in the FP: origin at the outflow tract of the RV; 3) RBBB morphology with left axis deviation in the FP: origin at the posterior region of the left ventricle (LV). RBBB morphology with right axis deviation in the FP: origin at the anterior region of the LV. RESULTS: Twenty three (92%) patients showed > 720 isolated ventricular ectopic beats/24 h. Paired ventricular response was detected in 18 (72%) patients and non-sustained VT in 15 (60%). HRECG was positive in six (24%) patients and Qtc interval was prolonged in 13 (52%). RV was the site of origin of the ventricular ectopic beats in 85% of the patients (outflow: 85%; inflow: 15%). Only five (20%) patients had arrhythmias from the LV. CONCLUSION: There was a high incidence of ventricular arrhythmias with a low incidence of positive HRECG tests, suggesting that the mechanisms of the arrhythmias do not correlate with slow intramyocardial conduction. It was noted a strong association between mitral valve prolapse, arrhythmogenic right ventricular disease and Qtc prolongation. It is possible that in some of this patients the finding could represent a global myocardial disease.