A comparison of four DNA extraction protocols for the analysis of urine from patients with visceral leishmaniasis

Introduction Polymerase chain reaction (PCR) may offer an alternative diagnostic option when clinical signs and symptoms suggest visceral leishmaniasis (VL) but microscopic scanning and serological tests provide negative results. PCR using urine is sensitive enough to diagnose human visceral leishmaniasis (VL). However, DNA quality is a crucial factor for successful amplification. Methods A comparative performance evaluation of DNA extraction methods from the urine of patients with VL using two commercially available extraction kits and two phenol-chloroform protocols was conducted to determine which method produces the highest quality DNA suitable for PCR amplification, as well as the most sensitive, fast and inexpensive method. All commercially available kits were able to shorten the duration of DNA extraction. Results With regard to detection limits, both phenol: chloroform extraction and the QIAamp DNA Mini Kit provided good results (0.1 pg of DNA) for the extraction of DNA from a parasite smaller than Leishmania (Leishmania) infantum (< 100fg of DNA). However, among 11 urine samples from subjects with VL, better performance was achieved with the phenol:chloroform method (8/11) relative to the QIAamp DNA Mini Kit (4/11), with a greater number of positive samples detected at a lower cost using PCR. Conclusion Our results demonstrate that phenol:chloroform with an ethanol precipitation prior to extraction is the most efficient method in terms of yield and cost, using urine as a non-invasive source of DNA and providing an alternative diagnostic method at a low cost.

Evaluation of parasitological examination, kDNA polymerase chain reaction and rK39-based immunochromatography for the diagnosis of visceral leishmaniasis in seropositive dogs from the screening-culling program in Brazil

Introduction Dogs play a primary role in the zoonotic cycle of visceral leishmaniasis (VL). Therefore, the accurate diagnosis of infected dogs, primarily asymptomatic dogs, is crucial to the efficiency of VL control programs. Methods We investigated the agreement of four diagnostic tests for canine visceral leishmaniasis (CVL): parasite detection, either after myeloculture or by direct microscopic examination of tissue imprints; kinetoplast-deoxyribonucleic acid-polymerase chain reaction (kDNA-PCR); and an immunochromatographic test (ICT). An enzyme-linked immunosorbent assay (ELISA) and an indirect immunofluorescence test (IFAT), both of which were adopted as part of the screening-culling program in Brazil, were used as reference tests. Our sample set consisted of 44 seropositive dogs, 25 of which were clinically asymptomatic and 19 were symptomatic for CVL according to ELISA-IFAT. Results The highest and lowest test co-positivities were observed for ICT (77.3%) and myeloculture (58.1%), respectively. When analyzed together, the overall percentage of co-positive tests was significantly higher for the symptomatic group compared to the asymptomatic group. However, only ICT was significantly different based on the results of a separate analysis per test for each group of dogs. The majority (93.8%) of animals exhibited at least one positive test result, with an average of 2.66 positive tests per dog. Half of the symptomatic dogs tested positive for all four tests administered. Conclusions The variability between test results reinforces the need for more efficient and reliable methods to accurately diagnose canine VL, particularly in asymptomatic animals.

Detection of canine visceral leishmaniasis by conjunctival swab PCR

Abstract: INTRODUCTION: Conjunctival swab PCR was evaluated as a tool to diagnose visceral leishmaniasis in dogs. METHODS: Conjunctival swab PCR was compared to indirect immunofluorescence antibody test and blood PCR. RESULTS: Indirect immunofluorescence was significantly correlated with conjunctival swab PCR (p < 0.05), but not with blood PCR (p > 0.05). In addition, conjunctival swab PCR was significantly associated with presence of clinical symptoms (p < 0.05), whereas blood PCR was associated with absence of clinical symptoms (p < 0.05). CONCLUSIONS: Results indicate that conjunctival swab PCR is useful in epidemiological surveys of canine visceral leishmaniasis.

REVISÃO DO GÊNERO NEOTROPICAL Acanthocera MACQUART (DIPTERA: TABANIDAE)

O gênero AcanthoceraMacquart é redefinido e apresentado lista sinonimica, diagnose, figuras e chave para fêmeas das 28 espécies. O subgênero NothocanthoceraFairchild do gênero Dichelaceraé transferido para Acanthocera,exceto D. melanopteraHine, e descrita uma espécie: A. distincta,sp. n. O subgênero MimodynerusEnderlein é sinonimizado sob A. {Acanthocera). A. bequaertiFairchüd & Aitken é revalidada. A. luizi(Enderlein) é sinonimizada sob A. coarctata(Wiedemann). A fêmea de A. polistiformisFairchild é descrita. Um novo lectótipo é designado para A. tenuicornis.

Anatomia foliar de pedra-hume-caá (Myrcia sphaerocarpa, Myrcia guianensis, Eugenia punicifolia - Myrtaceae)

As folhas de Myrcia multiflora (Lam.) DC, Myrcia guianensis (Aubl.) Urb. e Eugenia punicifolia (H. B. K.) DC. são anatômica c morfologicamentc descritas. As espécies revelaram diversos elementos histológicos universais para a família, bem como peculiares das mesmas e importantes para orientar a diagnose desses vegetais e para o reconhecimento de fraudes, que ocorrem através de substituições intencionais ou não. Entre os elementos histológicos característicos, destacam-se: ornamentações de cutícula foliar, tipos de estômatos, tipos de contornos celulares das células epidérmicas em vista facial, tipo e abundância relativa de inclusões celulares inorgânicas.

Geological characteristics of the nesting areas of the giant Amazon river turtle (Podocnemis expansa) in the Crixás-Açu river in Goiás State, Brazil

A large population of the giant Amazon river turtle (Podocnemis expansa) nests along the beaches of the Crixás-Açu River in the central western region of Brazil. In spite of the existence of several point bars in the area, only a selected group of beaches is used for nesting by P. expansa. Geological aspects, such as river width and depth, beach height above 200 cm with sandy sediments, were indispensable for the choice of these nesting sites. The relatively reduced dimensions of the point bars and the great number of turtles, which nest in the same local, contributed to the existence of a high nest concentration. The rapid rise of the Crixás-Açu River caused the flooding of the beaches and the drowning of hatchlings and embryos. It is estimated that nearly all the nests were lost. The height of the nesting place and the time of flooding related to the incubation period are decisive in embryo survivorship. The Retiro, Júnior, Assombrado and Limoeiro beaches, which are situated at heights of 183 to 310 cm, were inundated on 8 November 2000. The Barreira Branca beach, with a height of up to 380 cm was completely inundated on 13 December 2000. All of these beaches were flooded before the hatchlings emerged.

Relações nutricionais log-transformadas para avaliação nutricional de cupuaçueiros comerciais

A transformação logarítmica das relações bivariadas no cálculo das normas e dos índices do sistema integrado de diagnose e recomendação de nutrientes (DRIS) tem sido sugerida como uma forma de melhorar a acurácia do sistema, principalmente por diminuir a inconsistência na distribuição de freqüência entre as formas de expressão direta e inversa de uma mesma relação. Neste sentido, o objetivo deste trabalho foi avaliar o uso de relações log-transformadas entre diferentes populações de referência. Amostras foliares de cupuaçu foram coletadas de 153 pomares comerciais, cuja idade das plantas variou de 5 a 18 anos, cultivados em monocultivo ou sistemas agroflorestais, obtendo-se para cada relação nutricional entre os nutrientes N, P, K, Ca, Mg, Fe, Cu, Zn, e Mn as normas DRIS bivariadas log-transformadas e não transformadas, obtidas para o conjunto da população e para condições específicas. Os resultados mostraram que as relações log-transformadas contribuem para uma maior consistência dos resultados entre as formas direta e inversa entre diferentes normas DRIS.

Acmella marajoensis G.A.R. Silva & J.U.M. Santos: uma nova espécie de Asteraceae para a Amazônia brasileira

Em decorrência dos estudos na subtribo Ecliptinae da Amazônia brasileira, é descrita uma nova espécie, Acmella marajoensis G.A.R. Silva & J.U.M. Santos. Até o presente momento, a espécie é considerada como endêmica da Ilha do Marajó. São apresentados diagnose em latim, descrição detalhada, comentários taxonômicos e ilustrações.

Níveis críticos e faixas de suficiência nutricional em laranjeira-pêra na Amazônia Central obtidas pelo método DRIS

A avaliação do estado nutricional da laranjeira depende da definição de valores de referência que sejam adequados para refletir suas condições nutricionais. Neste trabalho, objetivou-se determinar os valores de referências e avaliar o estado nutricional de laranjeiras-pêra em diversas glebas na Amazônia Central (municípios de Iranduba, Manacapuru, Manaus, Presidente Figueiredo e Rio Preto da Eva). Utilizou-se o Sistema Integrado de Diagnose e Recomendação de relações multivariadas (DRIS) para estabelecer os valores de referência nutricional. O diagnóstico nutricional de 120 glebas comerciais de laranjeiras-pêra, enxertadas em limoeiro cravo foram avaliadas pelas faixas de suficiência definidas a partir do conjunto de plantas nutricionalmente equilibradas. Para os macronutrientes, as faixas de suficiência nutricional foram (g kg-1): 28-30 (para nitrogênio, N); 1,6-1,7 (fósforo, P); 7-9 (potássio, K); 26-29 (cálcio, Ca); 3,4-4 (magnésio, Mg); 1,7-2 (enxofre, S) e para os micronutrientes (mg kg-1): 47-56 (boro, B); 8-10 (cobre, Cu); 84-93 (ferro, Fe); 12-13 (manganês, Mn); 14-16 (zinco, Zn). Para os macronutrientes, os níveis críticos foram (g kg-1): 28 (para N); 1,6 (P); 7 (K); 26 (Ca); 3,6 (Mg); 1,7 (S) e para os micronutrientes (mg kg-1): 47 (B); 8 (Cu); 84 (Fe); 12 (Mn); 14 para Zn. Padrões nutricionais obtidos pelo DRIS discordam das faixas de suficiência propostas pela literatura para maioria dos nutrientes. Em quase 50% das glebas monitoradas, P, K, Ca, S, B, Cu e Fe estão abaixo dos níveis críticos propostos neste trabalho. Isto sugere que os produtores de laranja na Amazônia Central deveriam atentar-se para estes elementos no planejamento das fertilizações.

Taxonomic relevance of seed and seedling morphology in two Amazonian species of Entada (Leguminosae)

We carried out a comparative morphological study to evaluate the taxonomic value of seed and seedling traits of Entada polystachya and E. simplicata (Leguminosae, Mimosoideae, Mimoseae). Seeds of both species were collected from wild populations in Roraima State, Brazil. Our results show consistent quantitative and qualitative differences between these species regarding their seeds and seedlings. Testa fracture lines, funiculus impression, lens characters, radicle shape and length, and cotyledon lobes length, seedling morphology group, number of pinnae at first node, and hypocotyl length permit a straightforward distinction of E. polystachya from E. simplicata, supporting a recent treatment of the latter taxon as a distinct species, endemic to Roraima State, northern Amazonia, Brazil.

Comorbidities of mental and behavioral disorders in chemically dependent patients in different periods of abstinence

OBJECTIVE: To assess the frequency of comorbidities of mental and behavioral disorders (CMBD) in psychoactive substance (PAS)-dependent patients with different periods of abstinence cared for at Alcohol and Other Drug Psychosocial Care Centers (CAPS-ad). METHOD: All patients under treatment in the two CAPS-ad of the city of Uberlândia-MG, between April and September 2010, were consecutively assessed. The ICD-10 symptom checklist was used to diagnose CMBD; additional information was obtained from interviews and medical records. The patients were divided according to duration of abstinence: < 1 week (Group 1); 1-4 weeks (Group 2); and > 4 weeks (Group 3). RESULTS: Of all patients assessed, 62.8% were diagnosed with CMBD, which were more frequent (p < 0.05) in Group 1 (72%) than Group 3 (54.2%), and both groups were similar to Group 2 (61%). Depressive and anxiety disorders were more frequent among patients of Group 1. Mood disorders were more frequent (p < 0.05) in women [22/34 (65%) vs. 54/154 (35.1%)], whereas psychotic disorders were more frequent (p = 0.05) in men [16/154 (10.4%) vs. 0]. The presence of CMBD was associated with more severe clinical conditions. CONCLUSIONS: The higher frequency of diagnosis of CMBD in patients of Group 1 may have resulted from the difficulties in distinguishing mental disorders that are due to PAS intoxication or withdrawal from those that are not. However, to make the diagnosis of CMBD, even during detoxification, can increase the likelihood of better response to treatment.

Anomalous origin of the left coronary artery from the right pulmonary artery with intramural aortic trajectory. Clinicosurgical diagnostic implications

OBJECTIVE: Anomalous origin of the left coronary artery from the right pulmonary artery (AOLCARPA), is a rare entity that is usually associated with other defects. Of the 20 cases of AOLCARPA reported in the literature, 14 (70%) had associations. We describe four patients with AOLCARPA without associated defects, but with a peculiar intramural aortic trajectory. METHODS: Fifty-five patients with anomalous origin of the left coronary artery were operated upon at INCOR-FMUSP. Four of the patients had the anomalous origin from the right pulmonary artery (RPA) without associated defects but with intramural aortic trajectory. Clinical and laboratory examinations were analyzed, as well as surgical findings. RESULTS: All patients had congestive heart failure (CHF) and 3 also had angina pectoris. Two patients had a murmur of mitral regurgitation, signs of myocardial infarction on the ECG and cardiomegaly. The shortening fraction varied from 9% to 23%. The hemodynamic study confirmed the diagnosis of anomalous origin of the coronary artery, but the intramural trajectory and the origin from the RPA were established only at surgery. In 3 patients, the technique of side-to-side anastomosis was performed with a good outcome. One patient, who underwent end-to-side anastomosis, died 6 months after the surgery. CONCLUSION: Association with other defects usually occurs in the AOLCARPA, and the intramural aortic trajectory is difficult to clinically diagnose but easy to surgically correct.

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.

Methodological Gaps in Left Atrial Function Assessment by 2D Speckle Tracking Echocardiography

Abstract The assessment of left atrial (LA) function is used in various cardiovascular diseases. LA plays a complementary role in cardiac performance by modulating left ventricular (LV) function. Transthoracic two-dimensional (2D) phasic volumes and Doppler echocardiography can measure LA function non‑invasively. However, evaluation of LA deformation derived from 2D speckle tracking echocardiography (STE) is a new feasible and promising approach for assessment of LA mechanics. These parameters are able to detect subclinical LA dysfunction in different pathological condition. Normal ranges for LA deformation and cut-off values to diagnose LA dysfunction with different diseases have been reported, but data are still conflicting, probably because of some methodological and technical issues. This review highlights the importance of an unique standardized technique to assess the LA phasic functions by STE, and discusses recent studies on the most important clinical applications of this technique.

Carlos Chagas Discoveries as a Drop Back to Scientific Construction of Chronic Chagas Heart Disease

Abstract The scientific construction of chronic Chagas heart disease (CCHD) started in 1910 when Carlos Chagas highlighted the presence of cardiac arrhythmia during physical examination of patients with chronic Chagas disease, and described a case of heart failure associated with myocardial inflammation and nests of parasites at autopsy. He described sudden cardiac death associated with arrhythmias in 1911, and its association with complete AV block detected by Jacquet's polygraph as Chagas reported in 1912. Chagas showed the presence of myocardial fibrosis underlying the clinical picture of CCHD in 1916, he presented a full characterization of the clinical aspects of CCHD in 1922. In 1928, Chagas detected fibrosis of the conductive system, and pointed out the presence of marked cardiomegaly at the chest X-Ray associated with minimal symptomatology. The use of serological reaction to diagnose CCHD was put into clinical practice in 1936, after Chagas' death, which along with the 12-lead ECG, revealed the epidemiological importance of CCHD in 1945. In 1953, the long period between initial infection and appearance of CCHD was established, whereas the annual incidence of CCHD from patients with the indeterminate form of the disease was established in 1956. The use of heart catheterization in 1965, exercise stress testing in 1973, Holter monitoring in 1975, Electrophysiologic testing in 1973, echocardiography in 1975, endomyocardial biopsy in 1981, and Magnetic Resonance Imaging in 1995, added to the fundamental clinical aspects of CCHD as described by Carlos Chagas.