KSG estimation of reconstruction delay to detect vocal disorders in nonlinear dynamical analysis

Abstract Introduction This research investigates the applicability of a relatively new estimator of mutual information, KSG estimator, to find the reconstruction delay of phase space in dynamical systems. There are evidences that the KSG estimator is more accurate than the naive method commonly used. Methods In this paper we estimated mutual information between the voice signals and their delayed versions, with KSG method. The voice signals were obtained from a disordered voice database. Then, we found the reconstruction delay where mutual information reached its first minimum. We applied the encountered value of reconstruction delay in linear discriminant analysis, in order to discriminate between healthy and pathological voices or to discriminate between pathologies. Discrimination between voice pathologies using nonlinear measurements is still not much explored. Moreover, in this paper we used a single nonlinear measurement: reconstruction delay. Results The results show that the reconstruction delay obtained with KSG method has increased classification rates in most cases, in terms of accuracy, sensitivity and specificity, when compared to the naive estimator usually adopted. Conclusion The KSG estimator is a promising technique to improve the diagnosis of voice related pathologies.

The Echinococcus canadensis (G7) genome: a key knowledge of parasitic platyhelminth human diseases.

BACKGROUND: The parasite Echinococcus canadensis (G7) (phylum Platyhelminthes, class Cestoda) is one of the causative agents of echinococcosis. Echinococcosis is a worldwide chronic zoonosis affecting humans as well as domestic and wild mammals, which has been reported as a prioritized neglected disease by the World Health Organisation. No genomic data, comparative genomic analyses or efficient therapeutic and diagnostic tools are available for this severe disease. The information presented in this study will help to understand the peculiar biological characters and to design species-specific control tools. RESULTS: We sequenced, assembled and annotated the 115-Mb genome of E. canadensis (G7). Comparative genomic analyses using whole genome data of three Echinococcus species not only confirmed the status of E. canadensis (G7) as a separate species but also demonstrated a high nucleotide sequences divergence in relation to E. granulosus (G1). The E. canadensis (G7) genome contains 11,449 genes with a core set of 881 orthologs shared among five cestode species. Comparative genomics revealed that there are more single nucleotide polymorphisms (SNPs) between E. canadensis (G7) and E. granulosus (G1) than between E. canadensis (G7) and E. multilocularis. This result was unexpected since E. canadensis (G7) and E. granulosus (G1) were considered to belong to the species complex E. granulosus sensu lato. We described SNPs in known drug targets and metabolism genes in the E. canadensis (G7) genome. Regarding gene regulation, we analysed three particular features: CpG island distribution along the three Echinococcus genomes, DNA methylation system and small RNA pathway. The results suggest the occurrence of yet unknown gene regulation mechanisms in Echinococcus. CONCLUSIONS: This is the first work that addresses Echinococcus comparative genomics. The resources presented here will promote the study of mechanisms of parasite development as well as new tools for drug discovery. The availability of a high-quality genome assembly is critical for fully exploring the biology of a pathogenic organism. The E. canadensis (G7) genome presented in this study provides a unique opportunity to address the genetic diversity among the genus Echinococcus and its particular developmental features. At present, there is no unequivocal taxonomic classification of Echinococcus species; however, the genome-wide SNPs analysis performed here revealed the phylogenetic distance among these three Echinococcus species. Additional cestode genomes need to be sequenced to be able to resolve their phylogeny.

Estudo comparativo dos métodos diagnósticos para Leishmaniose Visceral em cães oriundos de Ilha Solteira, SP / Comparative study of diagnostic methods for visceral leishmaniasis in dogs from Ilha Solteira, SP

O objetivo da presente pesquisa foi avaliar comparativamente os métodos diagnósticos da Leishmaniose Visceral Canina (LVC), utilizando-se o ensaio imunoenzimático indireto (ELISA), a reação de imunofluorescência indireta (RIFI), a histoquímica (HE) e a imunoistoquímica (IMIQ) em tecidos de órgãos, como o baço, linfonodo e fígado. Além disso, a Reação em Cadeia pela Polimerase (PCR) das amostras de sangue e dos tecidos foi utilizada para comparar e confirmar os diagnósticos negativos e não conclusivos pelos métodos acima. Para esse estudo, foram utilizados 34 cães com diferentes sintomas da LVC, classificados em polissintomáticos, oligossintomáticos e assintomáticos e eutanasiados no Centro de Controle de Zoonoses (CCZ) de Ilha Solteira, SP. Os índices de positividade para os testes ELISA, IMIQ, RIFI e HE foram de 65,0, 62,0, 56,0 e 56,0 por cento, respectivamente, sendo a maior positividade detectada nos cães polissintomáticos (92,0 por cento), seguida pelos oligossintomáticos (57,0 por cento) e assintomáticos (12,5 por cento). A PCR confirmou os resultados positivos pelas outras técnicas e ainda detectou DNA do parasita nos tecidos de 100 por cento dos cães negativos e em 89,0 por cento dos suspeitos, elevando para 97,0 por cento a positividade. Em conclusão, a PCR demonstrou ser o método mais sensível e preciso para o diagnóstico definitivo da LVC.

The purpose of the present work was a comparative study of diagnostic methods for Canine Visceral Leishmaniasis (CVL) using serological methods, enzyme-linked immunosorbent assay (ELISA) and indirect fluorescent antibody test (IFAT), histochemical (HE) and immunohistochemical (IMHC) tests using spleen, lymph node and liver canine tissues. In addition, Polymerase Chain Reaction (PCR) was done in blood and in tissues in order to compare and confirm no conclusive and negative diagnosis by the methods above. For this study, 34 dogs were divided according to clinical signs in asymptomatic, oligosymptomatic and polisymptomatic Leishmania-infected dogs euthanized by Zoonotic Disease Control Center (CCZ) from Ilha Solteira, SP, Brazil. The positivism indexes of ELISA, IMHC, IFAT and HE were 65.0, 62.0, 56.0 and 56.0 percent, respectively with the highest numbers of positive dogs in polisymptomatic (92.0 percent) followed by oligosymptomatic (57.0 percent) and asymptomatic dogs (12.5 percent). Furthermore, PCR confirmed the positive results and detected DNA in tissues from 100 percent of negative dogs and 89.0 percent suspects raising the animal positivism index up to 97.0 percent. In conclusion, PCR was the most sensitive and a valuable method for a definitive CVL diagnosis.

Diagnóstico da Leishmaniose Visceral Canina pelas técnicas de imunoistoquímica e PCR em tecidos cutâneos em associação com a RIFI e ELISA-teste / Canine Visceral Leishmaniasis diagnosis by immunohistochemistry and PCR in skin tissues in association with RIFI and ELISA-test

O objetivo deste trabalho foi avaliar as técnicas de imunoistoquímica (IMIQ) e de PCR (Reação em Cadeia da Polimerase) em tecidos cutâneos para o diagnóstico da Leishmaniose Visceral Canina (LVC) e compará-los com os exames parasitológicos em tecidos corados histoquimicamente (hematoxilina-eosina, HE) e com testes sorológicos, como a Reação de Imunofluorescência Indireta (RIFI) e ensaio imunoenzimático (ELISA). Dos 34 cães naturalmente infectados, classificados em assintomáticos, oligossintomáticos e polissintomáticos, foram coletadas amostras de pele sadia ou com lesão para a realização da IMIQ, HE e PCR. Não somente peles lesionadas (56,5 por cento), mas também sadias (31,8 por cento) encontravam-se positivas pela IMIQ, confirmadas posteriormente pela PCR em 97,8 por cento das amostras. No grupo assintomático, 87,5 por cento estavam negativos pelos testes sorológicos, mas positivos em 50 por cento dos casos pela IMIQ e 100 por cento pela PCR. Entre os oligossintomáticos, 100 por cento, 85,7 por cento e 28,6 por cento encontravam-se positivos, respectivamente, pela PCR, sorologia e IMIQ. Os cães polissintomáticos eram 91,7 por cento soropositivos e tinham parasitas na pele. Em geral, a técnica PCR teve maior positividade (100 por cento). A eficiência dos testes variou de acordo com a evolução da doença, demonstrando a necessidade da associação de técnicas, usando-se IMIQ para confirmação da sorologia e a PCR apenas nos casos suspeitos após a IMIQ. Dessa forma, pode-se aumentar os níveis de positividade e contribuir para o controle desta zoonose.

The purpose of the present study was to evaluate the immunohistochemistry (IMHC) and PCR (Polymerase Chain Reaction) tests for Canine Visceral Leishmaniasis (CVL) diagnosis and compare the results with serological tests such as the indirect fluorescence antibody test (IFAT), ELISA and a parasitological test (microscopic direct examination of the parasite stained with haematoxylin and eosin - HE). For this study, samples of healthy or lesion skin tissues were obtained from 34 CVL naturally infected dogs classified in three groups: asymptomatic, oligosymptomatic and polisymptomatic. Not only lesion (56.5 percent) but also healthy skins (31.8 percent) were positives by IMHC and confirmed by PCR in 97.8 percent of skin samples. In asymptomatic group, 87.5 percent dogs were negatives by serological tests, but positives by IMHC in 50 percent and by PCR in 100 percent. In oligosymptomatic group, 100 percent, 85.7 percent and 28.6 percent of dogs were positives, respectively by PCR, serological and IMHC tests. In addition, 91.7 percent of polisymptomatic dogs were serum positive and had intact parasites in the skin. In general, PCR showed higher positivity (100 percent). The efficiency of each test varied with the evolution of the disease. IMHC may be used to confirm the results of the serology and PCR in inconclusive cases after HE and IMHC. The association of techniques proposed in this study may increase the positivity and contributed to the control of this canine disease.

Diagnóstico da Leishmaniose Visceral Canina pelas técnicas de imunoistoquímica e PCR em tecidos cutâneos em associação com a RIFI e ELISA-teste / Canine Visceral Leishmaniasis diagnosis by immunohistochemistry and PCR

O objetivo deste trabalho foi avaliar as técnicas de imunoistoquímica (IMIQ) e de PCR (Reação em Cadeia da Polimerase) em tecidos cutâneos para o diagnóstico da Leishmaniose Visceral Canina (LVC) e compará-los com os exames parasitológicos em tecidos corados histoquimicamente (hematoxilina-eosina, HE) e com testes sorológicos, como a Reação de Imunofluorescência Indireta (RIFI) e ensaio imunoenzimático (ELISA). Dos 34 cães naturalmente infectados, classificados em assintomáticos, oligossintomáticos e polissintomáticos, foram coletadas amostras de pele sadia ou com lesão para a realização da IMIQ, HE e PCR. Não somente peles lesionadas (56,5%), mas também sadias (31,8%) encontravam-se positivas pela IMIQ, confirmadas posteriormente pela PCR em 97,8% das amostras. Nogrupo assintomático, 87,5% estavam negativos pelos testes sorológicos, mas positivos em 50% dos casos pela IMIQ e 100% pela PCR. Entre os oligossintomáticos, 100%, 85,7% e 28,6% encontravam-se positivos, respectivamente, pela PCR, sorologia e IMIQ. Os cães polissintomáticos eram 91,7% soropositivos e tinham parasitas na pele. Em geral, a técnica PCR teve maior positividade (100%). A eficiência dos testes variou de acordo com a evolução da doença, demonstrando a necessidade da associação de técnicas, usando-se IMIQ para confirmação da sorologia e a PCR apenas nos casos suspeitos apos a IMIQ. Dessa forma, pode-se aumentar os níveis de positividade e contribuir para o controle desta zoonose.

The purpose of the present study was to evaluate the immunohistochemistry (IMHC) and PCR (Polymerase Chain Reaction) tests for Canine Visceral Leishmaniasis (CVL) diagnosis and compare the results with serological tests such as the indirect fluorescence antibody test (IFAT), ELISA and a parasitological test (microscopic direct examination of the parasite stained with haematoxylin and eosin - HE). For this study, samples of healthy or lesion skin tissues were obtained from 34 CVL naturally infected dogs classified in three groups: asymptomatic, oligosymptomatic and polisymptomatic. Not only lesion (56.5%) but also healthy skins (31.8%) were positives by IMHC and confirmed by PCR in 97.8% of skin samples. In asymptomatic group, 87.5% dogs were negatives by serological tests, but positivesby IMHC in 50% and by PCR in 100%. In oligosymptomatic group, 100%, 85.7% and 28.6% of dogs were positives, respectively by PCR, serological and IMHC tests. In addition, 91.7% of polisymptomatic dogs were serum positive and had intact parasites in the skin. In general, PCR showed higher positivity (100%). The efficiency of each test varied with the evolution of the disease. IMHC may be used to confirm the results of the serology and PCR in inconclusive cases after HE and IMHC. The association of techniques proposed in this study may increase the positivity and contributed to the control of this canine disease.

Estudo comparativo dos métodos diagnósticos para Leishmaniose Visceral em cães oriundos de Ilha Solteira, SP / Comparative study of diagnostic methods for visceral leishmaniasis in dogs from Ilha Solteira, SP

O objetivo da presente pesquisa foi avaliar comparativamente os métodos diagnósticos da Leishmaniose Visceral Canina (LVC), utilizando-se o ensaio imunoenzimático indireto (ELISA), a reação de imunofluorescência indireta (RIFI), a histoquímica (HE) e a imunoistoquímica (IMIQ) em tecidos de órgãos, como o baço, linfonodo e fígado. Além disso, a Reação em Cadeia pela Polimerase (PCR) das amostras de sangue e dos tecidos foi utilizada para comparar e confirmar os diagnósticos negativos e não conclusivos pelos métodos acima. Para esse estudo, foram utilizados 34 cães com diferentes sintomas da LVC, classificados em polissintomáticos, oligossintomáticos e assintomáticos e eutanasiados no Centro de Controle de Zoonoses (CCZ) de Ilha Solteira, SP. Os índices de positividade para os testes ELISA, IMIQ, RIFI e HE foram de 65,0, 62,0, 56,0 e 56,0%, respectivamente, sendo a maior positividade detectada nos cães polissintomáticos (92,0%), seguida pelos oligossintomáticos (57,0%) e assintomáticos (12,5%). A PCR confirmou os resultados positivos pelas outras técnicas e ainda detectou DNA do parasita nos tecidos de 100% dos cães negativos e em 89,0% dos suspeitos, elevando para 97,0% a positividade. Em conclusão, a PCR demonstrou ser o método mais sensível e preciso para o diagnóstico definitivo da LVC.

The purpose of the present work was a comparative study of diagnostic methods for Canine Visceral Leishmaniasis (CVL) using serological methods, enzyme-linked immunosorbent assay (ELISA) and indirect fluorescent antibody test (IFAT), histochemical (HE) and immunohistochemical (IMHC) tests using spleen, lymph node and liver canine tissues. In addition, Polymerase Chain Reaction (PCR) was done in blood and in tissues in order to compare and confirm no conclusive and negative diagnosis by the methods above. For this study, 34 dogs were divided according to clinical signs in asymptomatic, oligosymptomatic and polisymptomatic Leishmania-infected dogs euthanized by Zoonotic Disease Control Center (CCZ) from Ilha Solteira, SP, Brazil. The positivism indexes of ELISA, IMHC, IFAT and HE were 65.0, 62.0, 56.0 and 56.0%, respectively with the highest numbers of positive dogs in polisymptomatic (92.0%) followed by oligosymptomatic (57.0%) and asymptomatic dogs (12.5%). Furthermore, PCR confirmed the positive results and detected DNA in tissues from 100% of negative dogs and 89.0% suspects raising the animal positivism index up to 97.0%. In conclusion, PCR was the most sensitive and a valuable method for a definitive CVL diagnosis.

[Parasitological, immunohistochemical and histopathological study for Leishmania chagasi detection in splenic tissues of dogs with visceral leishmaniasis]. / Exames parasitológicos, imunoistoquímicos e histopatológicos para detecção de Leishania chagasi em tecidos esplênicos de cães com leishmaniose visceral.

The purpose of this work was a Canine Visceral Leishmaniasis--CVL study by parasitological direct examination of Leishmania (L.) chagasi (imprinting and histological), immunohistochemical test and histopathological analysis using spleen tissues from 34 dogs euthanized by the Zoonotic Disease Control Centre from Ilha Solteira, SP, Brazil. According to the clinical signs, the dogs were divided in three groups: asymptomatics (8 dogs), oligosymptomatics (17 dogs) and symptomatics (9 dogs). After the accomplishment of all diagnostic tests, 22 dogs were considered positives (64.7%) and 12 (35.3%) were negatives to CVL. From these positive dogs, 1/22 (4.5%) was asymptomatic, 12/22(54.5%) were oligosymptomatics and 8/22 (40.1%) were symptomatics. The histopathological study in spleen tissues from positive, especially symptomatic dogs, showed a diffuse chronic inflammation with thickness of capsular and trabecular regions and there was extensive morphologic alteration of the red and white pulp by the presence of abundant macrophages full with amastigotes, the granulomatous inflammatory reaction and haemorrhagic areas. The data of this work from histopathologic examination and direct microscopic visualization of L. (L.) chagasi showed that the spleen was an useful organ to collect sample tissues for CVL diagnosis. The immunostaining detected the highest number of positive dogs and were considered an important and conclusive method to be used in addition to parasitological methods for CVL, particularly in asymptomatic or oligosymptomatic dogs.

Exames parasitológicos, imunoistoquímicos e histopatológicos para detecção de Leishmania chagasi em tecidos esplênicos de cães com leishmaniose visceral / Parasitological, immunohistochemical and histopathological study for Leishmania chagasi detection in spleenic tissues of dogs with visceral leishmaniasis

O propósito do presente trabalho foi o estudo da Leishmaniose Visceral Canina - LVC por meio de métodos parasitológicos e imunoistoquímicos para a detecção de formas amastigotas de Leishmania (L.) chagasi em baço, além de descrever a histopatologia das lesões esplênicas em 34 cães, com diferentes manifestações clínicas da LVC, eutanasiados pelo Centro de Controle de Zoonoses de Ilha Solteira, SP. Esses animais foram examinados clinicamente antes da eutánásia e de acordo com os sinais clínicos da LVC, foram classificados em três grupos: assintomáticos (8 cães), oligossintomáticos (17 cães) e sintomáticos (9 cães). Após a realização desses exames, dos 34 cães, 22 (64,7 por cento) estavam positivos e 12 (35,3 por cento) negativos. Desses cães positivos, 1/22 (4,5 por cento) era assintomático, 12/22 (54,5 por cento) eram oligossintomáticos e 9/22 (40,1 por cento) sintomáticos. Pela histopatologia, os cães, especialmente os sintomáticos apresentavam o baço com inflamação crônica e espessamento na região capsular e trabecular, além de extensa alteração morfológica na polpa vermelha e branca pela presença de grande quantidade de macrófagos repletos de amastigotas, pela reação granulomatosa inflamatória e pelas áreas hemorrágicas. Os exames histopatológicos e a detecção microscópica direta da L. (L.) chagasi revelaram que o baço é um órgão útil para auxiliar no diagnóstico da LVC. A coloração imunoistoquímica foi a que detectou o maior número de tecidos esplênicos positivos com amastigotas, além de elucidar os casos suspeitos pelos exames parasitológicos, principalmente, nos animais assintomáticos ou oligossintomáticos.

The purpose of this work was a Canine Visceral Leishmaniasis - CVL study by parasitological direct examination of Leishmania (L.) chagasi (imprinting and histological), immunohistochemical test and histopathological analysis using spleen tissues from 34 dogs euthanized by the Zoonotic Disease Control Centre from Ilha Solteira, SP, Brazil. According to the clinical signs, the dogs were divided in three groups: asymptomatics (8 dogs), oligosymptomatics (17 dogs) and symptomatics (9 dogs). After the accomplishment of all diagnostic tests, 22 dogs were considered positives (64.7 percent) and 12 (35.3 percent) were negatives to CVL. From these positive dogs, 1/22 (4.5 percent) was asymptomatic, 12/22 (54.5 percent) were oligosymptomatics and 8/22 (40.1 percent) were symptomatics. The histopathological study in spleen tissues from positive, especially symptomatic dogs, showed a diffuse chronic inflammation with thickness of capsular and trabecular regions and there was extense morphologic alteration of the red and white pulp by the presence of abundant macrophages full with amastigotes, the granulomatous inflammatory reaction and haemorragic areas. The data of this work from histopathologic examination and direct microcopic visualization of L. (L.) chagasi showed that the spleen was an useful organ to collect sample tissues for CVL diagnosis. The immunostaining detected the highest number of positive dogs and were considered an important and conclusive method to be used in addition to parasitological methods for CVL, particularly in asymptomatic or oligosymptomatic dogs.

Composição e preferência por microhábitat de imaturos de Odonata (Insecta) em um trecho de baixada do Rio Ubatiba, Maricá-RJ, Brasil / Composition and microhabitat preferences of Odonata (Insecta) immatures in a lowland section of the Rio Ubatiba, Maricá-RJ, Brazil

No período de maio de 1999 a maio de 2000 foram efetuadas coletas de imaturos de Odonata (Insecta), em sete diferentes substratos, ao longo de 50 metros de um trecho do Rio Ubatiba, Maricá, Rio de Janeiro, Brasil. Um total de 1.279 larvas de Odonata foi coligido e identificado em 27 espécies de cinco famílias. Acanthagrion lancea (Selys, 1876), Hetaerina auripennis (Burmeister, 1839), Micrathyria hesperis Ris, 1911 e Telebasis filiola (Perty, 1834) foram as espécies mais numerosas. O substrato com o maior número de indivíduos foi "vegetação em áreas de remanso". Grande parte das espécies estudadas demonstrou preferência por algum dos substratos. Dythemis multipunctata Kirby,1894, Erythrodiplax sp., M. hesperis, T. filiola, A. lancea, Erythemis sp., Coryphaeschna adnexa (Hagen, 1861) e H. auripennis demonstraram preferência por substratos orgânicos, enquanto que Brechmorhoga sp., B. praeCOX (Hagen, 1869) e Progomphus complicatus (Selys, 1854), por inorgânicos.