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1.
Viruses ; 14(9)2022 08 23.
Article En | MEDLINE | ID: mdl-36146654

Monkeypox infection is rapidly spreading across the world. Despite the increasing number of cases, only a few reports have been published, and most are on people living without HIV. We report here the first two cases of monkeypox infection in Bahia, Brazil, one of them in a person living with HIV, on stable treatment. Both cases had a similar evolution, with a limited number of lesions and mild symptoms, with a complete recovery after 7-10 days. The potential route of transmission was via oral sex for the first case and was undefined for the second one. Both cases were confirmed through detection of the viral genome by PCR, and the partial sequence of the first case indicates the infection was caused by the West African clade. These cases confirm that monkeypox infection is currently being transmitted in Brazil and that people living with HIV on stable treatment are not likely to present a more severe form of monkeypox.


HIV Infections , Mpox (monkeypox) , Brazil/epidemiology , HIV Infections/complications , Humans , Mpox (monkeypox)/diagnosis , Mpox (monkeypox)/epidemiology , Monkeypox virus/genetics , Polymerase Chain Reaction
2.
Einstein (Sao Paulo) ; 18: eAO4209, 2020.
Article En, Pt | MEDLINE | ID: mdl-36083851

OBJECTIVE: To determine the association between the presence of periodontal pathogens and low birthweight. METHODS: This observational and case-control study consisted of mothers of infants weighing <2,500g (Group A), and mothers of newborns weighing ≥2,500g (Group B), born at Hospital da Mulher in Feira de Santana (BA), Brazil. A semi-structured questionnaire covering demographic data, gestational history and aspects related to general and oral health was employed postpartum. Following a complete periodontal examination, biofilm samples were collected at six sites in the mouth. The participants were further categorized in terms of presence or absence of periodontitis. Differences between the groups were determined using Pearson's χ 2 test, odds ratio, and confidence intervals were obtained using the Mantel-Haenszel test. RESULTS: Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Treponema denticola, Tannerella forsythia and Prevotella intermedia were detected by polymerase chain reaction. A total of 303 postpartum women were evaluated, 224 (73.9%) in Group B. Statistically significant differences between the groups were found for age, body mass index and history of previous low birthweight babies. Group A had a higher frequency of periodontitis (33.34%) than Group B (16.22%). P. gingivalis and P. intermedia were detected more frequently among women with periodontitis (74.19% and 88.70%, respectively). CONCLUSION: In this population, there was no association between the presence of maternal periodontal pathogens and the occurrence of low birthweight infants.

3.
J Periodontol ; 91(1): 46-54, 2020 01.
Article En | MEDLINE | ID: mdl-31342509

BACKGROUND: Periodontitis, an inflammatory disease of multibacterial etiology that affects the protective and supporting tissues surrounding teeth, can influence the course of respiratory diseases, such as asthma, due to epithelial alterations arising from inflammatory and immunological processes, bronchial remodeling, or by the aspiration of pathogenic colonizers found in periodontal pockets. This study evaluated the levels of periodontal pathogens Prevotella intermedia, Porphyromonas gingivalis, Treponema denticola, Tannerella forsythia, and Aggregatibacter actinomycetemcomitans in the subgingival biofilm of individuals with and without severe asthma. METHODS: A case-control study enrolling 457 individuals (220 with asthma and 237 without asthma) was conducted at the Program for Control of Asthma in Bahia (ProAR) Clinic located in Salvador, Bahia, Brazil. A structured questionnaire was used to obtain data on sociodemographic, health status, and lifestyle habits. A clinical periodontal assessment was performed, including bleeding on probing, probing depth, and clinical attachment level. Subgingival biofilm was collected at the deepest site of each sextant, and bacterial DNA was extracted. Quantitative real-time PCR analysis was performed to detect and relatively quantify periodontopathogens in the biofilm. RESULTS: Statistically significant positive associations were found between periodontitis and severe asthma, (odds ratio [OR]adjusted] : 4.00; 95% confidence interval [CI]: 2.26 to 7.10). High levels of P. intermedia were found in association with the presence of severe asthma (ORadjusted : 2.64; 95% CI: 1.62 to 4.39; P < 0.01). CONCLUSIONS: The present results suggest that periodontitis and P. intermedia are associated with severe asthma. However, the functional consequences of this dysbiosis upon asthma susceptibility and its phenotypes remain unclear.


Asthma , Periodontitis , Aggregatibacter actinomycetemcomitans , Bacteroides , Brazil , Case-Control Studies , Humans , Porphyromonas gingivalis , Prevotella intermedia , Treponema denticola
4.
Einstein (Säo Paulo) ; 18: eAO4209, 2020. tab, graf
Article En | LILACS-Express | LILACS | ID: biblio-1133781

ABSTRACT Objective To determine the association between the presence of periodontal pathogens and low birthweight. Methods This observational and case-control study consisted of mothers of infants weighing <2,500g (Group A), and mothers of newborns weighing ≥2,500g (Group B), born at Hospital da Mulher in Feira de Santana (BA), Brazil. A semi-structured questionnaire covering demographic data, gestational history and aspects related to general and oral health was employed postpartum. Following a complete periodontal examination, biofilm samples were collected at six sites in the mouth. The participants were further categorized in terms of presence or absence of periodontitis. Differences between the groups were determined using Pearson's χ 2 test, odds ratio, and confidence intervals were obtained using the Mantel-Haenszel test. Results Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Treponema denticola, Tannerella forsythia and Prevotella intermedia were detected by polymerase chain reaction. A total of 303 postpartum women were evaluated, 224 (73.9%) in Group B. Statistically significant differences between the groups were found for age, body mass index and history of previous low birthweight babies. Group A had a higher frequency of periodontitis (33.34%) than Group B (16.22%). P. gingivalis and P. intermedia were detected more frequently among women with periodontitis (74.19% and 88.70%, respectively). Conclusion In this population, there was no association between the presence of maternal periodontal pathogens and the occurrence of low birthweight infants.


RESUMO Objetivo Determinar a associação entre a presença de patógenos periodontais e o baixo peso ao nascer. Métodos Trata-se de estudo observacional e caso controle. Participaram mães de bebês com peso <2.500g (Grupo A) e de recém-nascidos com peso ≥2.500g (Grupo B) nascidos no Hospital da Mulher, em Feira de Santana, (BA). Dados demográficos, história gestacional e aspectos relacionados à saúde geral e bucal foram obtidos por meio de questionário semiestruturado, aplicado no período pós-parto. Após exame periodontal completo, as amostras de biofilme foram coletadas em seis locais na boca. As participantes foram também categorizadas quanto à presença ou ausência de periodontite. As diferenças entre os grupos foram obtidas pelo teste do χ 2 de Pearson, odds ratio e os intervalos de confiança pelo teste de Mantel-Haenszel. Resultados Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Treponema denticola, Tannerella forsythia e Prevotella intermedia foram detectados por reação em cadeia de polimerase. Avaliaram-se 303 mulheres pós-parto, sendo 224 (73,9%) do Grupo B. Diferenças estatisticamente significativas entre os grupos foram observadas quanto à idade, ao índice de massa corporal e a história de partos prévios com bebês de baixo peso ao nascer. O Grupo A apresentou maior frequência de periodontite (33,34%) do que o Grupo B (16,22%). P. gingivalis e P. intermedia foram detectados com maior frequência em mulheres com periodontite (74,19% e 88,70%, respectivamente). Conclusão Nessa população, não houve associação entre a presença de patógenos periodontais maternos e a ocorrência de bebês com baixo peso ao nascer.

5.
AMB Express ; 8(1): 89, 2018 May 30.
Article En | MEDLINE | ID: mdl-29846821

Caseous lymphadenitis (LC) is a chronic contagious disease caused by Corynebacterium pseudotuberculosis, which mainly affects goats and sheep. Vaccination is an effective but not yet well-established method, partly due to a lack of knowledge surrounding the most effective immunoprotective components. The present study aimed to quantify and compare the in vivo expression of genes pld (phospholipase D), cpp (CP40), nanH (neuraminidase H), sodC (superoxide dismutase C) and spaC (adhesin) using qRT-PCR, with the respective expression in vitro. Caseous material of abscesses removed from five animals was cultured, with colonies suggestive of C. pseudotuberculosis identified. RNA extraction was performed on these samples, as well as on the respective pellets derived from liquid cultures brain heart infusion. After evaluating RNA integrity, complementary DNA was synthesized, followed by the relative quantification each of the genes of interest. Mean mRNA expression of the five genes found in abscesses and in cultures differed significantly, with respective values of: nanH 811.50 ± 198.27 and 359.35 ± 75.45 (p = 0.009); cpp 856.31 ± 385.11 and 154.54 ± 94.34 (p = 0.0039); plD 922.70 ± 450.73 and 212.41 ± 153.10 (p = 0.016); sodC 1,293.53 ± 564.75 and 223.63 ± 145.58 (p = 0.016); spaC 1,157.10 ± 525.13 and 214.26 ± 125.70 (p = 0,016). Expression was observed to be 6-8 times higher in abscesses than in cultures, Indicative that is a genetic expression of the in vitro bacterium exists, yet in vivo has a greater magnitude corroborating to one of these virulence factors in the pathogenesis of LC.

6.
J Hepatol ; 68(6): 1114-1122, 2018 06.
Article En | MEDLINE | ID: mdl-29428874

BACKGROUND & AIMS: All known hepatitis B virus (HBV) genotypes occur in humans and hominoid Old World non-human primates (NHPs). The divergent woolly monkey HBV (WMHBV) forms another orthohepadnavirus species. The evolutionary origins of HBV are unclear. METHODS: We analysed sera from 124 Brazilian monkeys collected during 2012-2016 for hepadnaviruses using molecular and serological tools, and conducted evolutionary analyses. RESULTS: We identified a novel orthohepadnavirus species in capuchin monkeys (capuchin monkey hepatitis B virus [CMHBV]). We found CMHBV-specific antibodies in five animals and high CMHBV concentrations in one animal. Non-inflammatory, probably chronic infection was consistent with an intact preCore domain, low genetic variability, core deletions in deep sequencing, and no elevated liver enzymes. Cross-reactivity of antisera against surface antigens suggested antigenic relatedness of HBV, CMHBV, and WMHBV. Infection-determining CMHBV surface peptides bound to the human HBV receptor (human sodium taurocholate co-transporting polypeptide), but preferentially interacted with the capuchin monkey receptor homologue. CMHBV and WMHBV pseudotypes infected human hepatoma cells via the human sodium taurocholate co-transporting polypeptide, and were poorly neutralised by HBV vaccine-derived antibodies, suggesting that cross-species infections may be possible. Ancestral state reconstructions and sequence distance comparisons associated HBV with humans, whereas primate hepadnaviruses as a whole were projected to NHP ancestors. Co-phylogenetic analyses yielded evidence for co-speciation of hepadnaviruses and New World NHP. Bayesian hypothesis testing yielded strong support for an association of the HBV stem lineage with hominoid ancestors. Neither CMHBV nor WMHBV was likely the ancestor of the divergent human HBV genotypes F/H found in American natives. CONCLUSIONS: Our data suggest ancestral co-speciation of hepadnaviruses and NHP, and an Old World origin of the divergent HBV genotypes F/H. The identification of a novel primate hepadnavirus offers new perspectives for urgently needed animal models of chronic hepatitis B. LAY SUMMARY: The origins of HBV are unclear. The new orthohepadnavirus species from Brazilian capuchin monkeys resembled HBV in elicited infection patterns and could infect human liver cells using the same receptor as HBV. Evolutionary analyses suggested that primate HBV-related viruses might have emerged in African ancestors of New World monkeys millions of years ago. HBV was associated with hominoid primates, including humans and apes, suggesting evolutionary origins of HBV before the formation of modern humans. HBV genotypes found in American natives were divergent from those found in American monkeys, and likely introduced along prehistoric human migration. Our results elucidate the evolutionary origins and dispersal of primate HBV, identify a new orthohepadnavirus reservoir, and enable new perspectives for animal models of hepatitis B.


Cebus/virology , Evolution, Molecular , Hepatitis B virus/genetics , Hepatitis B virus/isolation & purification , Orthohepadnavirus/genetics , Orthohepadnavirus/isolation & purification , Amino Acid Sequence , Animals , Bayes Theorem , Brazil , Genetic Speciation , Genome, Viral , Hepatitis B/veterinary , Hepatitis B/virology , Hepatitis B Antigens/chemistry , Hepatitis B Antigens/genetics , Hepatitis B Antigens/immunology , Hepatitis B virus/classification , Host Microbial Interactions/genetics , Humans , Models, Genetic , Monkey Diseases/virology , Organic Anion Transporters, Sodium-Dependent/physiology , Orthohepadnavirus/classification , Phylogeny , Primates/virology , Receptors, Virus/physiology , Symporters/physiology , Virus Internalization
7.
Infect Drug Resist ; 10: 201-207, 2017.
Article En | MEDLINE | ID: mdl-28740410

BACKGROUND AND AIMS: Resistance mutation analogs to nucleos(t)ides have been described in treatment-naïve patients with chronic hepatitis B (CHB), with clinical implications. The aim of this study was to investigate primary resistance mutations and genotypes circulating in patients naïve to chronic hepatitis B, in the Northern and Northeastern regions of Brazil. METHODS: We conducted a study of resistance mutations and genotypic characterization of hepatitis B virus (HBV) in 189 treatment-naïve patients chronically infected with HBV. RESULTS: Drug resistance-associated mutations located in the RT domain of the P gene (rtHBV) were found in 6% of the treatment-naïve patients from the Northeastern Region. The mutations were rtA194T, rtL180M + rtM204V, rtS202I, rtM204I, and rtA181S. No patient in the Northern Region had the resistance mutation. In the gene S region, the frequency of vaccine escape mutations was 2.4% in the Northeastern Region and 8.6% in the Northern Region. CONCLUSION: This information before the start of treatment may contribute to clinical decision making, reducing treatment failure and the risk of progression to cirrhosis and hepatocellular carcinoma for CHB.

8.
J Med Virol ; 89(10): 1811-1816, 2017 10.
Article En | MEDLINE | ID: mdl-28500726

The Brazilian public health system (SUS) has provided antiviral drugs for chronic hepatitis B treatment for over 10 years, but a system for monitoring for drug-related resistance mutations is not available. Determine the presence of HBV mutations associated with resistance to nucleos(t)ide analogs among 81 patients with chronic HBV infection in Salvador-BA-Brazil. HBV-DNA was PCR amplified with primers deduced from the rt domain at the HBV P gene, the sequence extended 1032 bp (from amino acid 1 to 344-rt domain). Those sequences were submitted to the HBV drug resistance database to retrieve each mutation according to the genotype. HBV genotype A1 (85.2%) was the most prevalent, followed by genotype A2 (4.9%), F (6.2%), and C1, D2, and D4 (1.2% each). Six patients (7%) exhibited resistance mutations to LAM, ETV, and TDF: two with patterns L180M + M204V and four with other different patterns: L80I + L180M + M204I; L80V + L180M + M204V; M204I; A194T. All of these mutations were present in patients with genotype A (four A1 and two A2). In addition, four mutations in gene S (three cases with the sI195M mutation and one with the W196L mutation), were detected, corresponding to a rate of 6% of vaccine escape mutations. Althougth the small sample size, an association was found between the occurrence of HBV resistance mutations and HBeAg positivity, co-infection with HIV and a history of treatment for HBV and/or HIV.


Drug Resistance, Viral/genetics , Hepatitis B virus/drug effects , Hepatitis B virus/genetics , Hepatitis B, Chronic/virology , Adult , Antiviral Agents/therapeutic use , Brazil/epidemiology , Coinfection/epidemiology , Coinfection/virology , DNA, Viral/genetics , Genotype , Hepatitis B Vaccines , Hepatitis B virus/physiology , Hepatitis B, Chronic/drug therapy , Hepatitis B, Chronic/epidemiology , Humans , Lamivudine/therapeutic use , Male , Middle Aged , Mutation , Organophosphonates/therapeutic use
9.
Proc Natl Acad Sci U S A ; 110(40): 16151-6, 2013 Oct 01.
Article En | MEDLINE | ID: mdl-24043818

The hepatitis B virus (HBV), family Hepadnaviridae, is one of most relevant human pathogens. HBV origins are enigmatic, and no zoonotic reservoirs are known. Here, we screened 3,080 specimens from 54 bat species representing 11 bat families for hepadnaviral DNA. Ten specimens (0.3%) from Panama and Gabon yielded unique hepadnaviruses in coancestral relation to HBV. Full genome sequencing allowed classification as three putative orthohepadnavirus species based on genome lengths (3,149-3,377 nt), presence of middle HBV surface and X-protein genes, and sequence distance criteria. Hepatic tropism in bats was shown by quantitative PCR and in situ hybridization. Infected livers showed histopathologic changes compatible with hepatitis. Human hepatocytes transfected with all three bat viruses cross-reacted with sera against the HBV core protein, concordant with the phylogenetic relatedness of these hepadnaviruses and HBV. One virus from Uroderma bilobatum, the tent-making bat, cross-reacted with monoclonal antibodies against the HBV antigenicity determining S domain. Up to 18.4% of bat sera contained antibodies against bat hepadnaviruses. Infectious clones were generated to study all three viruses in detail. Hepatitis D virus particles pseudotyped with surface proteins of U. bilobatum HBV, but neither of the other two viruses could infect primary human and Tupaia belangeri hepatocytes. Hepatocyte infection occurred through the human HBV receptor sodium taurocholate cotransporting polypeptide but could not be neutralized by sera from vaccinated humans. Antihepadnaviral treatment using an approved reverse transcriptase inhibitor blocked replication of all bat hepadnaviruses. Our data suggest that bats may have been ancestral sources of primate hepadnaviruses. The observed zoonotic potential might affect concepts aimed at eradicating HBV.


Chiroptera/virology , Hepadnaviridae/genetics , Hepadnaviridae/pathogenicity , Zoonoses/virology , Animals , Base Sequence , Cell Line, Tumor , Cross Reactions/immunology , Electrophoresis, Polyacrylamide Gel , Enzyme-Linked Immunosorbent Assay , Fluorescent Antibody Technique , Genome/genetics , Hepatitis B virus/genetics , Hepatocytes/virology , Humans , Immunoblotting , In Situ Hybridization , Molecular Sequence Data , Sequence Analysis, DNA , Species Specificity , Tupaiidae
10.
J Gen Virol ; 94(Pt 9): 1984-1994, 2013 Sep.
Article En | MEDLINE | ID: mdl-23761408

Bats host a broad diversity of coronaviruses (CoVs), including close relatives of human pathogens. There is only limited data on neotropical bat CoVs. We analysed faecal, blood and intestine specimens from 1562 bats sampled in Costa Rica, Panama, Ecuador and Brazil for CoVs by broad-range PCR. CoV RNA was detected in 50 bats representing nine different species, both frugivorous and insectivorous. These bat CoVs were unrelated to known human or animal pathogens, indicating an absence of recent zoonotic spill-over events. Based on RNA-dependent RNA polymerase (RdRp)-based grouping units (RGUs) as a surrogate for CoV species identification, the 50 viruses represented five different alphacoronavirus RGUs and two betacoronavirus RGUs. Closely related alphacoronaviruses were detected in Carollia perspicillata and C. brevicauda across a geographical distance exceeding 5600 km. Our study expands the knowledge on CoV diversity in neotropical bats and emphasizes the association of distinct CoVs and bat host genera.


Chiroptera/virology , Coronavirus/classification , Coronavirus/isolation & purification , Genetic Variation , Americas , Animals , Blood/virology , Cluster Analysis , Coronavirus/genetics , Feces/virology , Intestines/virology , Molecular Sequence Data , Phylogeography , RNA, Viral/genetics , RNA-Dependent RNA Polymerase/genetics , Sequence Analysis, DNA
11.
J Periodontol ; 84(5): 650-5, 2013 May.
Article En | MEDLINE | ID: mdl-22769440

BACKGROUND: In chronic periodontitis (CP), the gene polymorphism of interleukin-6 (IL-6) to 174C/G has been associated with the altered production of this cytokine. The aim of this pilot study is to compare the allelic and genotypic frequencies in patients with CP with control individuals without periodontitis (NP) and to measure the production of IL-6 by whole blood cells stimulated with Porphyromonas gingivalis HmuY protein. METHODS: DNA was isolated from peripheral blood cells of 49 patients with CP and 60 control individuals classified as NP, and genotyping was performed by polymerase chain reaction using sequence-specific primers. Whole blood cells from 29 patients with CP and 30 control individuals were stimulated for 48 hours with HmuY, and IL-6 levels were measured using enzyme-linked immunosorbent assay. RESULTS: The proportion of individuals carrying the G allele at position -174 of the IL-6 gene was higher in the group with CP (85.7%) than in the normal control group (73.3%; P <0.03). P. gingivalis HmuY-induced production of IL-6 was higher in the group with CP (P <0.05). CONCLUSIONS: Our findings suggest that P. gingivalis HmuY may be associated with increased IL-6 production during CP. Furthermore, patients with periodontitis and individuals with higher HmuY-induced production of IL-6 show a high frequency of the G allele at position -174.


Bacterial Proteins/physiology , Chronic Periodontitis/genetics , Chronic Periodontitis/microbiology , Interleukin-6/biosynthesis , Interleukin-6/genetics , Porphyromonas gingivalis , Adult , Bacterial Outer Membrane Proteins/physiology , Case-Control Studies , Chronic Periodontitis/metabolism , Female , Gene Frequency , Host-Pathogen Interactions , Humans , Logistic Models , Male , Middle Aged , Pilot Projects , Polymorphism, Genetic , Porphyromonas gingivalis/chemistry , Porphyromonas gingivalis/physiology , ROC Curve , Statistics, Nonparametric
12.
Emerg Infect Dis ; 18(4): 656-9, 2012 Apr.
Article En | MEDLINE | ID: mdl-22469070

To determine possible cosavirus association with clinical disease, we used real-time reverse transcription PCR to test children and HIV-positive adults in Brazil with and without gastroenteritis. Thirteen (3.6%) of 359 children with gastroenteritis tested positive, as did 69 (33.8%) of 204 controls. Low prevalence, frequent viral co-infections, and low fecal cosavirus RNA concentrations argue against human pathogenicity.


Diarrhea/epidemiology , Gastroenteritis/epidemiology , Picornaviridae Infections/epidemiology , Picornaviridae/isolation & purification , Adult , Brazil/epidemiology , Case-Control Studies , Child , Cohort Studies , Coinfection , Diarrhea/virology , Feces/virology , Gastroenteritis/virology , Genes, Viral , Humans , Molecular Sequence Data , Picornaviridae/genetics , Picornaviridae Infections/virology , Prevalence , Sequence Analysis, DNA
13.
Nat Commun ; 3: 796, 2012 Apr 24.
Article En | MEDLINE | ID: mdl-22531181

The large virus family Paramyxoviridae includes some of the most significant human and livestock viruses, such as measles-, distemper-, mumps-, parainfluenza-, Newcastle disease-, respiratory syncytial virus and metapneumoviruses. Here we identify an estimated 66 new paramyxoviruses in a worldwide sample of 119 bat and rodent species (9,278 individuals). Major discoveries include evidence of an origin of Hendra- and Nipah virus in Africa, identification of a bat virus conspecific with the human mumps virus, detection of close relatives of respiratory syncytial virus, mouse pneumonia- and canine distemper virus in bats, as well as direct evidence of Sendai virus in rodents. Phylogenetic reconstruction of host associations suggests a predominance of host switches from bats to other mammals and birds. Hypothesis tests in a maximum likelihood framework permit the phylogenetic placement of bats as tentative hosts at ancestral nodes to both the major Paramyxoviridae subfamilies (Paramyxovirinae and Pneumovirinae). Future attempts to predict the emergence of novel paramyxoviruses in humans and livestock will have to rely fundamentally on these data.


Chiroptera/virology , Disease Reservoirs/virology , Mammals/virology , Paramyxoviridae Infections/virology , Paramyxoviridae/classification , Paramyxoviridae/isolation & purification , Animals , Dogs , Humans , Mice , Molecular Sequence Data , Paramyxoviridae/genetics , Phylogeny
14.
J Gen Virol ; 92(Pt 3): 564-71, 2011 Mar.
Article En | MEDLINE | ID: mdl-21123543

Due to high genome plasticity, the evolutionary fate and geographical history of picornaviruses is hard to follow. Here, we determined the complete coding sequences of eight human parechoviruses (HPeV) of types 1, 5 and 6 directly from clinical samples from Brazil. The capsid genes of these strains were not remarkably different from European, North American and Japanese HPeV. Full genome analysis revealed frequent intertypic recombination in the non-structural genome region. In addition, evidence of recombination between viruses of the same type in the capsid-encoding genome region among HPeV1 and HPeV4 was obtained. Bayesian phylogenetic analysis indicated that strains without evidence of recombination with each other in any genome region were separated by no more than 35 years of circulation. Interestingly, in the 3C gene, all Brazilian parechoviruses grouped together regardless of serotype. The most recent common ancestor of these strains dated back 108 years, suggesting long-term endemicity of this particular P3 genome lineage in South America. Our results support the idea that picornavirus replicative genes acquire capsid proteins introduced by new strains. Under certain epidemiological conditions, replicative genes may be maintained in circumscript geographical regions.


Evolution, Molecular , Genome, Viral , Parechovirus/genetics , Parechovirus/isolation & purification , Picornaviridae Infections/virology , Recombination, Genetic , Brazil , Capsid Proteins/genetics , Cluster Analysis , Humans , Molecular Sequence Data , Parechovirus/classification , Phylogeny , RNA, Viral/genetics , Sequence Analysis, DNA , Sequence Homology , Viral Nonstructural Proteins/genetics
16.
J Gen Virol ; 91(Pt 6): 1418-27, 2010 Jun.
Article En | MEDLINE | ID: mdl-20130131

This study identified the complete genomic sequence of four type 2 and type 3 human Saffold-like cardioviruses (SLCVs) isolated in Germany and Brazil. The secondary structures of the SLCV internal ribosome entry sites (IRESs) were deduced based on RNA base-pairing conservation and co-variation, using an established Theiler's murine encephalomyelitis virus (TMEV) IRES structure as a reference. The SLCV IRES was highly similar to that of TMEV, but motifs critical in TMEV for binding of the polypyrimidine tract-binding protein (PTB) were disrupted. In TMEV, corresponding alterations have been associated with reduced neurovirulence in mice. In the non-structural genome region, there was evidence of multiple intertypic recombination events between different SLCV types. Between viruses of the same type, recombination also occurred in the capsid-encoding genome region. There were apparently no recombination events between mouse TMEV and human SLCV. In another genus of the family Picornaviridae, Enterovirus, natural recombination occurs strictly within species and can serve as an additional criterion for delimiting species. Accordingly, the results of this study suggest that SLCV and TMEV may represent distinct species within the genus Cardiovirus.


Cardiovirus/genetics , Evolution, Molecular , Genome, Viral , RNA, Viral/genetics , Sequence Analysis, DNA , Animals , Brazil , Cardiovirus/classification , Cardiovirus/isolation & purification , Cardiovirus Infections/virology , Germany , Humans , Mice , Molecular Sequence Data , Nucleic Acid Conformation , Recombination, Genetic , Theilovirus/genetics , Viral Nonstructural Proteins/genetics , Viral Structural Proteins/genetics
18.
Emerg Infect Dis ; 14(9): 1398-405, 2008 Sep.
Article En | MEDLINE | ID: mdl-18760006

Cardioviruses cause serious disease, mainly in rodents, including diabetes, myocarditis, encephalomyelitis, and multiple sclerosis-like disseminated encephalomyelitis. Recently, a human virus isolate obtained 25 years ago, termed Saffold virus, was sequenced and classified as a cardiovirus. We conducted systematic molecular screening for Saffold-like viruses in 844 fecal samples from patients with gastroenteritis from Germany and Brazil, across all age groups. Six cardioviruses were identified in patients <6 years of age. Viral loads were 283,305-5,044,412,175 copies/g of stool. Co-infections occurred in 4 of 6 children. No evidence for outbreak-like epidemic patterns was found. Phylogenetic analysis identified 3 distinct genetic lineages. Viral protein 1 amino acids were 67.9%-77.7% identical and had a distance of at least 39.4% from known cardioviruses. Because closely related strains were found on 2 continents, global distribution in humans is suspected. Saffold-like viruses may be the first human cardiovirus species to be identified.


Cardiovirus Infections/epidemiology , Cardiovirus Infections/virology , Cardiovirus/genetics , Adolescent , Adult , Aged , Aged, 80 and over , Base Sequence , Brazil/epidemiology , Cardiovirus/classification , Child , Child, Preschool , Feces/virology , Gastroenteritis/epidemiology , Gastroenteritis/virology , Germany/epidemiology , Humans , Infant , Infant, Newborn , Middle Aged , Oligonucleotides , RNA, Viral , Reverse Transcriptase Polymerase Chain Reaction
19.
FEBS J ; 275(19): 4773-85, 2008 Oct.
Article En | MEDLINE | ID: mdl-18721136

The protonation state of residues around the Q(o) binding site of the cytochrome bc(1) complex from Paracoccus denitrificans and their interaction with bound quinone(s) was studied by a combined electrochemical and FTIR difference spectroscopic approach. Site-directed mutations of two groups of conserved residues were investigated: (a) acidic side chains located close to the surface and thought to participate in a water chain leading up to the heme b(L) edge, and (b) residues located in the vicinity of this site. Interestingly, most of the mutants retain a high degree of catalytic activity. E295Q, E81Q and Y297F showed reduced stigmatellin affinity. On the basis of electrochemically induced FTIR difference spectra, we suggest that E295 and D278 are protonated in the oxidized form or that their mutation perturbs protonated residues. Mutations Y302, Y297, E81 and E295, directly perturb signals from the oxidized quinone and of the protein backbone. By monitoring the interaction with the inhibitor stigmatellin for the wild-type enzyme at various redox states, interactions of the bound stigmatellin with amino acid side chains such as protonated acidic residues and the backbone were observed, as well as difference signals arising from the redox active inhibitor itself and the replaced quinone. The infrared difference spectra of the above Q(o) site mutations in the presence of stigmatellin confirm the previously established role of E295 as a direct interaction partner in the enzyme from P.denitrificans as well. The protonated residue E295 is proposed to change the hydrogen-bonding environment upon stigmatellin binding in the oxidized form, and is deprotonated in the reduced form. Of the residues located close to the surface, D278 remains protonated and unperturbed in the oxidized form but its frequency shifts in the reduced form. The mechanistic implications of our observations are discussed, together with previous inhibitor binding data, and referred to the published X-ray structures.


Electron Transport Complex III/genetics , Paracoccus denitrificans/metabolism , Binding Sites/genetics , Mutagenesis, Site-Directed , Oxidation-Reduction , Polyenes/pharmacology , Spectroscopy, Fourier Transform Infrared , Tyrosine/metabolism , Ubiquinone/analogs & derivatives , Ubiquinone/metabolism
20.
J Trace Elem Med Biol ; 19(2-3): 183-9, 2005.
Article En | MEDLINE | ID: mdl-16325534

Royal jelly from Apis mellifera is a highly active natural biological substance and is probably one of the most interesting raw substances in natural product chemistry. Trace elements play a key role in the biomedical activities associated with royal jelly, as these elements have a multitude of known and unknown biological functions. For this reason concentrations of 28 trace (Al, Ba, Sr, Bi, Cd, Hg, Pb, Sn, Te, Tl, W, Sb, Cr, Ni, Ti, V, Co, Mo) and mineral (P, S, Ca, Mg, K, Na, Zn, Fe, Cu, Mn) elements were systematically investigated in botanically and geologically defined royal jelly samples. In addition, concentrations of 14 trace elements were measured in the associated honey samples--honey being the precursor of royal jelly. Concentrations of K, Na, Mg, Ca, P, S, Cu, Fe, Zn, Al, Ba and Sr in royal jelly were determined by inductively coupled plasma optical emission spectroscopy (ICP-OES), while concentrations of Bi, Cd, Hg, Pb, Sn, Te, Tl, W, Sb, Cr, Mn, Ni, Ti, V, Co and Mo in royal jelly were determined by double focusing magnetic sector field inductively coupled plasma mass spectrometry (SF-ICP-MS). In the honey samples, trace and mineral element concentrations strongly depended on botanical and geological origin, and substantial variation was found. In contrast, the concentrations of trace and mineral elements were highly constant in the associated royal jelly samples. The most important results were the homeostatic adjustments of trace and mineral element concentrations in royal jelly. This effect was evidently produced in the endocrine glands of nurse bees, which are adapted for needs of bee larvae. In conclusion, this research yielded a surprising and completely new finding--that royal jelly, as a form of lactation on the insect level, shows the same homeostatic adjustment as mammalian and human breast milk.


Chemistry Techniques, Analytical/methods , Fatty Acids/metabolism , Minerals/analysis , Trace Elements , Animals , Bees/metabolism , Elements , Homeostasis , Mass Spectrometry , Temperature
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