Carbon nanotubes functionalized with sodium hyaluronate restore bone repair in diabetic rat sockets.

OBJECTIVE: We evaluated the effects of sodium hyaluronate (HY) and carbon nanotubes functionalized with HY (HY-CNT) on bone repair in the tooth sockets of diabetic rats. MATERIALS AND METHODS: Diabetes was induced by streptozotocin (50 mg kg(-1) i.v.), and the sockets were divided into normal control, diabetic control, diabetic treated with HY (1%), and diabetic treated with HY-CNT (100 µg ml(-1)) groups. The sockets were analyzed according to the percentage of bone formation and the number of cell nuclei. RESULTS: The percentage of bone trabeculae was lower in diabetic control animals (11.16 ± 5.10% vs 41.92 ± 6.34% in normal animals) after 14 days. Treating diabetic animals with HY or HY-CNT significantly increased the percentage of neoformed trabeculae (HY: 29.43 ± 3.29%; HY-CNT: 36.90 ± 3.07%). Moreover, the sockets of diabetic animals had an increased number of cell nuclei and HY or HY-CNT reduced this parameter. CONCLUSION: Our results indicate that HY and HY-CNT restore bone repair in the tooth sockets of diabetic rats, suggesting that these biomaterials are potential adjuvant therapies for the management of diabetes.

Egg production and life history of Alona guttata Sars, 1862 (Cladocera, Chydoridae): implications for colonization of temporary ponds.

Cladocerans are a diverse group of species that show rapid responses to changes in environmental conditions. This adaptive capacity has important implications for egg production and life cycle, especially in transitory environments such as temporary waterbodies. The present study investigated the life history and egg production of Alona gutatta Sars, 1862 (Crustacea, Cladocera), an abundant and frequent species from a high-altitude temporary pond (Lagoa Seca, Minas Gerais, Brazil). Newly hatched neonates were monitored in relation to time of maturation, number of eggs produced per female and time of survival. Neonates required a mean of 8 days to mature. A. guttata survived for a mean of 30.9 ± 8.1 days and produced 2 eggs per brood, generating a mean of 10.95 ± 6.41 neonates during the entire life cycle. The rapid development, short time to produce eggs and long life cycle are important adaptations to the adverse environmental conditions of temporary aquatic environments, which can contribute to the rapid colonization of Alona guttata in transitory ecosystems.

Highly efficient siRNA delivery system into human and murine cells using single-wall carbon nanotubes.

Development of RNA interference (RNAi) technology utilizing short interfering RNA sequences (siRNA) has focused on creating methods for delivering siRNAs to cells and for enhancing siRNA stability in vitro and in vivo. Here, we describe a novel approach for siRNA cellular delivery using siRNA coiling into carboxyl-functionalized single-wall carbon nanotubes (SWCNTs). The CNT-siRNA delivery system successfully demonstrates nonspecific toxicity and transfection efficiency greater than 95%. This approach offers the potential for siRNA delivery into different types of cells, including hard-to-transfect cells, such as neuronal cells and cardiomyocytes. We also tested the CNT-siRNA system in a non-metastatic human hepatocellular carcinoma cell line (SKHep1). In all types of cells used in this work the CNT-siRNA delivery system showed high efficiency and apparent no side effects for various in vitro applications.

A methodological proposal for the analysis of Cladocera (Crustacea, Branchiopoda) subfossils from sediments of temporary aquatic ecosystems.

The main objective of the present study was to promote some adaptations to the standard methodology used in the analysis of Cladocera subfossils. The samples used in this study were collected at Lagoa dos Coutos, a temporary lake located on an outcrop with iron-rich duricrust in the Serra do Gandarela (Minas Gerais, Brazil). The sediment from this pond, typical of temporary ecosystems in the region, is characterized by rigid concretions that are difficult to dissolve in aqueous media, making it difficult to visualize and analyze subfossils taxa. Sediment samples were prepared according to the standard methodology used in paleolimological studies and the methodological adaptations proposed in the present study, which included the addition of 1 mg of Kochia scoparia, a palynological marker used to estimate subfossil density, as well as the addition of hydrofluoric acid (HF) to oxidize silicate materials. The samples prepared according to the new methodological procedure improved Cladocera subfossil visualization, facilitating the taxonomic analysis of the species. The samples submitted to the standard methodological procedure had a significantly higher mean of sediment particles (mean of 84.2 sediment particles) and 70% more residues compared to the samples prepared with the newly proposed methodology (mean of 22.1 sediment particles). Absolute data on the temporal variation of the density of Cladocera subfossils was obtained by using the palinological marker K. scoparia, which showed higher densities of subfossils in more recent, humid periods (112,000 subfossils/cm3 at 5 cm, equivalent to 3,500 years BP). The results indicate that the new methodology can contribute to advances in paleolimnological studies of temporary aquatic ecosystems, whose lake histories are rarely investigated, despite their ecological relevance as ecosystems that indicate environmental and climatic changes.

Echovirus 30 detection in an outbreak of acute myalgia and rhabdomyolysis, Brazil 2016-2017.

OBJECTIVES: To describe an outbreak of acute myalgia accompanied by elevated levels of muscle enzymes that occurred in the northeast region of Brazil from December 2016 through to May 2017. METHODS: Clinical data were analysed and laboratory tests were performed in 86 specimens obtained from 52 individuals with suspected acute myalgia. A broader reactive enterovirus real-time RT-PCR followed by a semi-nested PCR amplification of partial VP1 gene were performed to identify the causative agent. RESULTS: Eighty-six clinical samples were received in our laboratory during the myalgia outbreak. Median age of individuals was 39 years. Sudden acute myalgia and dark urine were the most common symptoms. Creatine phosphokinase levels were elevated with mean value ∼16 893 U/L. Human enterovirus was detected in 67% (58/86) of the patient's specimens (urine, serum, faeces and rectal swab). The enterovirus positivity per patient was 82.7% (43/52). Echovirus 30 (E-30) (82% of the typed specimens, 18/22; 76.4% (13/17) of the typed specimens per patient) was the main enterovirus identified. In addition to E-30, CV-A16 (1/22) and E-6 (3/22) were detected in 4% and 14% of the typed specimens, respectively. No deaths occurred. CONCLUSION: The 2016-2017 outbreak of acute myalgia that occurred in the northeast region of Brazil can be associated with E-30. Despite the clinical manifestations, a favourable outcome was observed for all patients.

Egg production and life history of Alona guttata Sars, 1862 (Cladocera, Chydoridae): implications for colonization of temporary ponds / Produção de ovos e história de vida de Alona guttata Sars, 1862 (Cladocera, Chydoridae): implicações para a colonização de lagoas temporárias

Cladocerans are a diverse group of species that show rapid responses to changes in environmental conditions. This adaptive capacity has important implications for egg production and life cycle, especially in transitory environments such as temporary waterbodies. The present study investigated the life history and egg production of Alona gutatta Sars, 1862 (Crustacea, Cladocera), an abundant and frequent species from a high-altitude temporary pond (Lagoa Seca, Minas Gerais, Brazil). Newly hatched neonates were monitored in relation to time of maturation, number of eggs produced per female and time of survival. Neonates required a mean of 8 days to mature. A. guttata survived for a mean of 30.9 ± 8.1 days and produced 2 eggs per brood, generating a mean of 10.95 ± 6.41 neonates during the entire life cycle. The rapid development, short time to produce eggs and long life cycle are important adaptations to the adverse environmental conditions of temporary aquatic environments, which can contribute to the rapid colonization of Alona guttata in transitory ecosystems.

Os cladóceros formam um grupo diverso de espécies que apresentam respostas rápidas às mudanças nas condições ambientais. Essa capacidade adaptativa tem implicações importantes para a produção de ovos e para o ciclo de vida, especialmente em ambientes transitórios, como corpos d’água temporários. O presente estudo investigou a história de vida e a produção de ovos da espécie Alona guttata Sars, 1862 (Crustacea, Cladocera) coletada em uma lagoa temporária de altitude (Minas Gerais, Brasil). Organismos recém eclodidos foram observados (em condições de laboratório) em relação ao tempo de maturação, número de ovos produzidos por fêmea e tempo de sobrevivência. Os neonatos levaram em média 8 dias para atingir o primeiro estágio de maturação. As fêmeas produziram 2 ovos por ninhada e geraram uma média de 10,95 ± 6,41 neonatos durante todo o ciclo de vida. A. guttata apresentou uma média de 30,9 ± 8,1 dias de sobrevivência. O rápido desenvolvimento, o pouco tempo para a produção de ovos e o ciclo de vida longo são adaptações importantes às condições ambientais adversas de ambientes aquáticos temporários, que podem contribuir para a rápida colonização de Alona guttata em ecossistemas transitórios.

Egg production and life history of Alona guttata Sars, 1862 (Cladocera, Chydoridae): implications for colonization of temporary ponds

Abstract Cladocerans are a diverse group of species that show rapid responses to changes in environmental conditions. This adaptive capacity has important implications for egg production and life cycle, especially in transitory environments such as temporary waterbodies. The present study investigated the life history and egg production of Alona gutatta Sars, 1862 (Crustacea, Cladocera), an abundant and frequent species from a high-altitude temporary pond (Lagoa Seca, Minas Gerais, Brazil). Newly hatched neonates were monitored in relation to time of maturation, number of eggs produced per female and time of survival. Neonates required a mean of 8 days to mature. A. guttata survived for a mean of 30.9 ± 8.1 days and produced 2 eggs per brood, generating a mean of 10.95 ± 6.41 neonates during the entire life cycle. The rapid development, short time to produce eggs and long life cycle are important adaptations to the adverse environmental conditions of temporary aquatic environments, which can contribute to the rapid colonization of Alona guttata in transitory ecosystems.

Resumo Os cladóceros formam um grupo diverso de espécies que apresentam respostas rápidas às mudanças nas condições ambientais. Essa capacidade adaptativa tem implicações importantes para a produção de ovos e para o ciclo de vida, especialmente em ambientes transitórios, como corpos d'água temporários. O presente estudo investigou a história de vida e a produção de ovos da espécie Alona guttata Sars, 1862 (Crustacea, Cladocera) coletada em uma lagoa temporária de altitude (Minas Gerais, Brasil). Organismos recém eclodidos foram observados (em condições de laboratório) em relação ao tempo de maturação, número de ovos produzidos por fêmea e tempo de sobrevivência. Os neonatos levaram em média 8 dias para atingir o primeiro estágio de maturação. As fêmeas produziram 2 ovos por ninhada e geraram uma média de 10,95 ± 6,41 neonatos durante todo o ciclo de vida. A. guttata apresentou uma média de 30,9 ± 8,1 dias de sobrevivência. O rápido desenvolvimento, o pouco tempo para a produção de ovos e o ciclo de vida longo são adaptações importantes às condições ambientais adversas de ambientes aquáticos temporários, que podem contribuir para a rápida colonização de Alona guttata em ecossistemas transitórios.

Egg production and life history of Alona guttata Sars, 1862 (Cladocera, Chydoridae): implications for colonization of temporary ponds / Produção de ovos e história de vida de Alona guttata Sars, 1862 (Cladocera, Chydoridae): implicações para a colonização de lagoas temporárias

Cladocerans are a diverse group of species that show rapid responses to changes in environmental conditions. This adaptive capacity has important implications for egg production and life cycle, especially in transitory environments such as temporary waterbodies. The present study investigated the life history and egg production of Alona gutatta Sars, 1862 (Crustacea, Cladocera), an abundant and frequent species from a high-altitude temporary pond (Lagoa Seca, Minas Gerais, Brazil). Newly hatched neonates were monitored in relation to time of maturation, number of eggs produced per female and time of survival. Neonates required a mean of 8 days to mature. A. guttata survived for a mean of 30.9 ± 8.1 days and produced 2 eggs per brood, generating a mean of 10.95 ± 6.41 neonates during the entire life cycle. The rapid development, short time to produce eggs and long life cycle are important adaptations to the adverse environmental conditions of temporary aquatic environments, which can contribute to the rapid colonization of Alona guttata in transitory ecosystems.

Os cladóceros formam um grupo diverso de espécies que apresentam respostas rápidas às mudanças nas condições ambientais. Essa capacidade adaptativa tem implicações importantes para a produção de ovos e para o ciclo de vida, especialmente em ambientes transitórios, como corpos d'água temporários. O presente estudo investigou a história de vida e a produção de ovos da espécie Alona guttata Sars, 1862 (Crustacea, Cladocera) coletada em uma lagoa temporária de altitude (Minas Gerais, Brasil). Organismos recém eclodidos foram observados (em condições de laboratório) em relação ao tempo de maturação, número de ovos produzidos por fêmea e tempo de sobrevivência. Os neonatos levaram em média 8 dias para atingir o primeiro estágio de maturação. As fêmeas produziram 2 ovos por ninhada e geraram uma média de 10,95 ± 6,41 neonatos durante todo o ciclo de vida. A. guttata apresentou uma média de 30,9 ± 8,1 dias de sobrevivência. O rápido desenvolvimento, o pouco tempo para a produção de ovos e o ciclo de vida longo são adaptações importantes às condições ambientais adversas de ambientes aquáticos temporários, que podem contribuir para a rápida colonização de Alona guttata em ecossistemas transitórios.

Acute haemorrhagic conjunctivitis outbreak in the city of Fortaleza, northeast Brazil.

BACKGROUND/AIM: Between February and May 2003 an epidemic of acute haemorrhagic conjunctivitis affected more than 200 000 people in all five geographic regions of Brazil (north, south, midwestern, southeast, and northeast). The aim was to identify the aetiological agent and to describe clinical aspects of this outbreak in a group of patients treated at the ophthalmology department of the Hospital Walter Cantídio (OD-HWC) at the Universidade Federal do Ceará, in the city of Fortaleza, capital of the state of Ceará, northeastern Brazil. METHODS: Conjunctival swabs were collected from patients who spontaneously went to the laboratory of virology. Specimens were inoculated in HEp-2 and RD cell lines. The viral isolation was confirmed by performing reverse transcriptase polymerase chain reaction and indirect immunofluorescence assay. RESULTS: Viral conjunctivitis was diagnosed in 56 patients but only 24 of them allowed the collection of samples. Of 24 conjunctival swabs tested, 11 were positive for a variant of coxsackie virus A24 (CA24v) and one of the isolates reacted with anti-adenovirus monoclonal antibodies. CONCLUSION: CA24v was confirmed as the aetiological agent of this outbreak of acute haemorrhagic conjunctivitis in the city of Fortaleza.

A methodological proposal for the analysis of Cladocera (Crustacea, Branchiopoda) subfossils from sediments of temporary aquatic ecosystems / Uma proposta metodológica para a análise de subfósseis de Cladocera (Crustacea, Branchiopoda) de sedimentos de ecossistemas aquáticos temporários

Abstract The main objective of the present study was to promote some adaptations to the standard methodology used in the analysis of Cladocera subfossils. The samples used in this study were collected at Lagoa dos Coutos, a temporary lake located on an outcrop with iron-rich duricrust in the Serra do Gandarela (Minas Gerais, Brazil). The sediment from this pond, typical of temporary ecosystems in the region, is characterized by rigid concretions that are difficult to dissolve in aqueous media, making it difficult to visualize and analyze subfossils taxa. Sediment samples were prepared according to the standard methodology used in paleolimological studies and the methodological adaptations proposed in the present study, which included the addition of 1 mg of Kochia scoparia, a palynological marker used to estimate subfossil density, as well as the addition of hydrofluoric acid (HF) to oxidize silicate materials. The samples prepared according to the new methodological procedure improved Cladocera subfossil visualization, facilitating the taxonomic analysis of the species. The samples submitted to the standard methodological procedure had a significantly higher mean of sediment particles (mean of 84.2 sediment particles) and 70% more residues compared to the samples prepared with the newly proposed methodology (mean of 22.1 sediment particles). Absolute data on the temporal variation of the density of Cladocera subfossils was obtained by using the palinological marker K. scoparia, which showed higher densities of subfossils in more recent, humid periods (112,000 subfossils/cm3 at 5 cm, equivalent to 3,500 years BP). The results indicate that the new methodology can contribute to advances in paleolimnological studies of temporary aquatic ecosystems, whose lake histories are rarely investigated, despite their ecological relevance as ecosystems that indicate environmental and climatic changes.

Resumo O principal objetivo do presente estudo foi promover algumas adaptações na metodologia padrão utilizadas para a análise de subfósseis de Cladocera. As amostras utilizadas nessa pesquisa foram coletadas na Lagoa dos Coutos, uma lagoa temporária encontrada em solo de afloramento ferruginoso na Serra do Gandarela (Minas Gerais, Brasil). O sedimento desta lagoa, típico de ecossistemas temporários da região, caracteriza-se por apresentar concreções rígidas de difícil dissolução em meio aquoso, que dificultam a visualização e análise taxonômica dos subfósseis. Amostras de sedimento foram preparadas de acordo com a metodologia padrão utilizada em estudos paleolimnológicos e as adaptações metodológicas propostas no presente estudo, que incluíram a adição de 1 mg de Kochia scoparia, um marcador palinológico utilizado para estimar a densidade de subfósseis, além da adição de ácido fluorídrico (HF) para oxidação de materiais silicáticos. As amostras preparadas de acordo com o novo procedimento metodológico apresentaram melhor visualização dos subfósseis de Cladocera, facilitando a análise taxonômica das espécies. As amostras submetidas ao procedimento metodológico padrão apresentaram uma média significativamente maior de partículas de sedimentos (média de 84,2 partículas de sedimento) e 70% mais resíduos em comparação com as amostras preparadas com a nova metodologia proposta (média de 22,1 partículas de sedimento). Dados absolutos sobre a variação temporal da densidade dos subfósseis de Cladocera foram obtidos a partir do uso do marcador palinológico K. scoparia, revelando densidades mais elevadas de subfósseis nos períodos recentes (112.000 subfósseis/cm3 nos 5 cm iniciais, equivalentes a 3.500 anos BP). Os resultados obtidos indicam que a nova metodologia pode contribuir para avanços em estudos paleolimnológicos de ecossistemas aquáticos temporários, raramente contemplados na reconstrução da história de lagos, apesar de sua relevância ecológica como ecossistemas sinalizadores de mudanças ambientais e climáticas.

Nucleotide sequences of the VP1 capsid proteins of wild poliovirus types 1 and 3 from epidemic areas of Brazil.

The nucleotide sequences encoding the capsid protein VP1 were determined for the wild polioviruses of serotypes 1 and 3 endemic to the northeastern region of Brazil. Compared with the corresponding Sabin vaccine strain sequences, the wild isolates differed at 20% (type 1) and 22% (type 3) of their nucleotide positions, and in 7% (type 1) and 11% (type 3) of their amino acid residues. The highest degree of amino acid heterogeneity occurred within the amino-terminal residues of the VP1 proteins. Intratypic amino acid differences also occurred in VP1 surface residues that form parts of antigenic sites for neutralizing antibodies.

Genomic characterization of type 3 polioviruses isolated from vaccine-associated poliomyelitis cases in Brazil.

Eight strains of P3/Sabin-related polioviruses were analyzed; four from persistent paralytic poliomyelitis cases classified as vaccine associated, one from a transient paralysis case classified as transverse myelitis, one from a transient paralysis case classified as Guillain-Barré syndrome, one from a transient facial paralysis case, and one from a healthy vaccine. The serotypes of the viral isolates were identified by the neutralization test with hyperimmune equine sera and the relationship of the isolates with the P3/Sabin strain was demonstrated by molecular hybridization of the viral RNA of the isolates with a P3/Sabin-specific probe. The P3/Sabin relationship was confirmed by PCR, using a pair of specific primers for P3/Sabin-related isolates. The available data indicate that a U-->C mutation at nucleotide 472 in the 5' noncoding region of the genome of the type 3 Sabin strain increases the neurovirulence of this strain and this mutation was observed in all type 3 isolates from vaccine-associated cases. These eight P3/Sabin-related isolates were partially sequenced in the 5' noncoding region and seven presented a U-->C mutation at nucleotide 472, except the isolate from a transient paralysis case classified as transverse myelitis, that maintained a U at nucleotide 472. Although this virus maintaining U at nucleotide 472 may not be the etiological agent of the disease, the possibility that the virus was the causative agent of the disease could not be ruled out.

Genomic characterization of type 2 polioviruses isolated from vaccine-associated cases in Brazil.

Twenty strains of P2/Sabin-related polioviruses isolated in Brazil were analyzed; ten from persistent paralytic poliomyelitis cases, three from suspected polio cases with transient paralysis, and seven from healthy contacts. The serotypes of the viral isolates were identified by the neutralization test with hyperimmune equine sera. The relationship of the isolates to the P2/Sabin strain was demonstrated by molecular hybridization and polymerase chain reaction (PCR). Partial sequencing demonstrated mutations at nucleotide 481 in the 5' noncoding region and at amino acid 143 of the capsid protein VP1 in most of these isolates from vaccine-associated cases in Brazil. These data support previous studies on the importance of mutations at these attenuated determinants in the establishment of the disease. However, the existence of isolates without mutations at these positions suggests that they are not essential. The results also strengthen the possibility of the participation of a mutation at nucleotide 398 in the establishment of the disease, and suggest that a mutation at nucleotide 491 or 500 may also be involved in this process. The isolates from healthy contacts presented the same mutations as the isolates from vaccine-associated cases with which they were in contact. This strengthens the observation that, although mutations in the genome of the P2/Sabin strain are important for the establishment of the disease, host factors are also involved.

Poliovirus type 1 isolated from a vaccine-associated case of paralytic poliomyelitis in Brazil.

This study reports a type 1 poliovirus strain isolated in Brazil from a case classified as vaccine-associated paralytic poliomyelitis (VAPP). After serotyping of the viral isolate with hyperimmune equine sera, PCR and molecular hybridization techniques characterized the strain as P1/Sabin-derived. The isolate was partially sequenced to identify mutations at nucleotides 480, 525 and 6203, which are important for reversion of the P1/Sabin strain to neurovirulence. In a recent study, a P1/Sabin-derived strain isolated from the central nervous system of a VAPP case did not mutate at these positions, but maintained 480-G and 525-U (and 6203-C), suggesting that these mutations are not essential for the occurrence of disease (Georgescu et al., (1994), Journal of Virology, 68: 8089-8101). Although the Brazilian strain also maintained 480-G and 525-U (and 6203-C) and was isolated from the stool, the possibility that this isolate invaded the central nervous system after replicating in the gut, causing the paralysis, cannot be ruled out. This is the first report of a type 1 VAPP case in Brazil, although some cases caused by type 2 and type 3 strains have been described.

Ultrastructural and immunocytochemical study on the infection of enterovirus 71 (EV 71) in rhabdomyosarcoma (RD) cells.

Rhabdomyosarcoma monolayers were inoculated with enterovirus 71 (EV 71) at 73 degrees C, sampled at intervals during the replicative cycle, and examined in thin sections by electron microscopy, using routine and immunoelectronmicroscopy with polyclonal antibodies against EV 71. The location of EV 71 or its precursors was followed during the viral replicative cycle. The earliest samples (3 h postinoculation) showed a cell shape change, from elongated to rounded. At 6 h postinoculation, the presence of early virus-induced vesicles developing within the cytoplasm was pointed out, although no virus particles were observed at these stages. At 12 and 20 h postinoculation, virus particles appeared in the cytoplasm. They were found free or in clusters in the cytoplasmic matrix, between the virus-induced vesicles. EV 71 particles were also occasionally observed in the form of paracrystalline arrays situated in the vesiculated areas. The immunolabel (gold beads) was found, initially, over dense cytoplasmic areas and in more advanced process at the vesiculated area and over the virus particles. Therefore the main cellular alterations observed in this infection were the shape change of the cell and the appearance of electron-dense areas (viroplasm) and smooth walled vesicles which are probably the site of the virus replication.

Enteroviruses isolated from patients with acute respiratory infections during seven years in Rio de Janeiro (1985-1991).

Enteroviruses were investigated in respiratory secretions collected from patients with acute respiratory infections (ARI) over a seven year period (1985-1991), as part of a longitudinal study of ARI aetiology. All the viruses that are most commonly associated with ARI were found in this study. Among the virus isolates, enteroviruses were only less frequent than respiratory syncytial viruses, adenoviruses and influenzaviruses. Forty five enterovirus samples were isolated from patients with either upper respiratory tract infections (URTI) or lower respiratory tract infections (LRTI). From these enterovirus isolates, thirty one samples were identified as poliovirus (n = 18) and non polio enterovirus (n = 13) by serum neutralization. Poliovirus were identified as type 1 and 2 and all of them were vaccinal strains. From thirteen non polio enterovirus, twelve were identified as echovirus serotypes 1, 2, 7, 11, 19 and 31. The remainder was identified as coxsackievirus B4.

Genomic characterization of type 1 Sabin-related polioviruses isolated in Brazil.

Eight strains of P1/Sabin-derived polioviruses isolated in Brazil from paralysis cases were analyzed. The serotypes of the viral isolates were identified by neutralization test with hyperimmune equine sera. The relationship of the isolates to the P1/Sabin strain was demonstrated by molecular hybridization and PCR. The isolates were partially sequenced with the objective of finding mutations at nucleotides (nt) 480 and 525 of the 5'-noncoding region (5' NCR) and at nt 6203 of the 3Dpol coding region (3Dpol), which are important for reversion towards neurovirulence. Four isolates from paralysis cases classified as Guillain Barré Syndrome (GBS; three with sequels) were analyzed; one presented G-->A (480) and C-->U (6203) mutations, one G-->A (480) mutation, one G-->A (480) and U-->C (525) mutations, and one did not mutate at the analyzed positions. Two isolates from transient facial paralysis cases were analyzed; one presented U-->C (525) mutation and the other G-->A (480) mutation. One isolate from a transient paralysis case classified as a neuroviral disease and one isolate from a paralysis case with sequels were analyzed and none mutated at the analyzed positions. Although the isolates may not be the causative agent of the disease, a temporal association between the isolation of the P1/Sabin-derived isolates and the disease was observed. The possibility that GBS and the facial paralysis were caused by these isolates could not be excluded.

A methodological proposal for the analysis of Cladocera (Crustacea, Branchiopoda) subfossils from sediments of temporary aquatic ecosystems

Abstract The main objective of the present study was to promote some adaptations to the standard methodology used in the analysis of Cladocera subfossils. The samples used in this study were collected at Lagoa dos Coutos, a temporary lake located on an outcrop with iron-rich duricrust in the Serra do Gandarela (Minas Gerais, Brazil). The sediment from this pond, typical of temporary ecosystems in the region, is characterized by rigid concretions that are difficult to dissolve in aqueous media, making it difficult to visualize and analyze subfossils taxa. Sediment samples were prepared according to the standard methodology used in paleolimological studies and the methodological adaptations proposed in the present study, which included the addition of 1 mg of Kochia scoparia, a palynological marker used to estimate subfossil density, as well as the addition of hydrofluoric acid (HF) to oxidize silicate materials. The samples prepared according to the new methodological procedure improved Cladocera subfossil visualization, facilitating the taxonomic analysis of the species. The samples submitted to the standard methodological procedure had a significantly higher mean of sediment particles (mean of 84.2 sediment particles) and 70% more residues compared to the samples prepared with the newly proposed methodology (mean of 22.1 sediment particles). Absolute data on the temporal variation of the density of Cladocera subfossils was obtained by using the palinological marker K. scoparia, which showed higher densities of subfossils in more recent, humid periods (112,000 subfossils/cm3 at 5 cm, equivalent to 3,500 years BP). The results indicate that the new methodology can contribute to advances in paleolimnological studies of temporary aquatic ecosystems, whose lake histories are rarely investigated, despite their ecological relevance as ecosystems that indicate environmental and climatic changes.

Resumo O principal objetivo do presente estudo foi promover algumas adaptações na metodologia padrão utilizadas para a análise de subfósseis de Cladocera. As amostras utilizadas nessa pesquisa foram coletadas na Lagoa dos Coutos, uma lagoa temporária encontrada em solo de afloramento ferruginoso na Serra do Gandarela (Minas Gerais, Brasil). O sedimento desta lagoa, típico de ecossistemas temporários da região, caracteriza-se por apresentar concreções rígidas de difícil dissolução em meio aquoso, que dificultam a visualização e análise taxonômica dos subfósseis. Amostras de sedimento foram preparadas de acordo com a metodologia padrão utilizada em estudos paleolimnológicos e as adaptações metodológicas propostas no presente estudo, que incluíram a adição de 1 mg de Kochia scoparia, um marcador palinológico utilizado para estimar a densidade de subfósseis, além da adição de ácido fluorídrico (HF) para oxidação de materiais silicáticos. As amostras preparadas de acordo com o novo procedimento metodológico apresentaram melhor visualização dos subfósseis de Cladocera, facilitando a análise taxonômica das espécies. As amostras submetidas ao procedimento metodológico padrão apresentaram uma média significativamente maior de partículas de sedimentos (média de 84,2 partículas de sedimento) e 70% mais resíduos em comparação com as amostras preparadas com a nova metodologia proposta (média de 22,1 partículas de sedimento). Dados absolutos sobre a variação temporal da densidade dos subfósseis de Cladocera foram obtidos a partir do uso do marcador palinológico K. scoparia, revelando densidades mais elevadas de subfósseis nos períodos recentes (112.000 subfósseis/cm3 nos 5 cm iniciais, equivalentes a 3.500 anos BP). Os resultados obtidos indicam que a nova metodologia pode contribuir para avanços em estudos paleolimnológicos de ecossistemas aquáticos temporários, raramente contemplados na reconstrução da história de lagos, apesar de sua relevância ecológica como ecossistemas sinalizadores de mudanças ambientais e climáticas.