RESUMO
A 15-year-old, mixed breed, male horse was attended with a history of multiple abscesses in the cervical region with a three-year evolution. Upon admission, three fistulous tracts with drainage of purulent secretions in the cervical region, low body score, restriction of cervical movements, and painful sensitivity to palpation were observed. The horse was diagnosed with osteomyelitis secondary to Streptococcus equi infection. The initial treatment was antibiotic therapy and local curative. Owing to the lack of response, surgical debridement was performed. An initial favorable response was observed; however, after 4 months, drainage recurred, and the animal was euthanized. A post-mortem computed tomography scan was performed to obtain details of the injury. Cervical osteomyelitis is rare, and its occurrence through hematogenous spread in adult horses and the tomographic findings had not been reported previously. The long period of evolution, difficulty in performing an aggressive debridement, and the presence of multi-drug resistant bacteria contributed to the negative outcome.(AU)
Um equino macho, sem raça definida, de 15 anos de idade, foi atendido com histórico de múltiplos abscessos cervicais com evolução de três anos. Na admissão, foram observados: três trajetos fistulosos com drenagem de material purulento na região cervical; baixo escore corporal; restrição de movimentos cervicais; e sensibilidade dolorosa à palpação da região. Foi diagnosticada osteomielite vertebral cervical secundária à infecção por Streptococcus equi. O tratamento inicial consistiu na administração de antibióticos e curativo local. Na ausência de resposta à terapia, realizou-se o debridamento cirúrgico. Inicialmente, obteve-se uma resposta favorável, entretanto, após quatro meses, houve recidiva da lesão e o animal foi submetido à eutanásia. Realizou-se tomografia computadorizada no post mortem para detalhamento da lesão. A osteomielite vertebral cervical é rara, e sua ocorrência por meio de disseminação hematógena em animais adultos não foi previamente reportada. O longo período de evolução, aliado à dificuldade de realização de um debridamento agressivo, e a característica multirresistente do agente etiológico contribuíram para o desfecho negativo do caso.(AU)
Assuntos
Animais , Osteomielite/veterinária , Infecções Estreptocócicas/complicações , Vértebras Cervicais/patologia , Streptococcus equi , CavalosRESUMO
A 15-year-old, mixed breed, male horse was attended with a history of multiple abscesses in the cervical region with a three-year evolution. Upon admission, three fistulous tracts with drainage of purulent secretions in the cervical region, low body score, restriction of cervical movements, and painful sensitivity to palpation were observed. The horse was diagnosed with osteomyelitis secondary to Streptococcus equi infection. The initial treatment was antibiotic therapy and local curative. Owing to the lack of response, surgical debridement was performed. An initial favorable response was observed; however, after 4 months, drainage recurred, and the animal was euthanized. A post-mortem computed tomography scan was performed to obtain details of the injury. Cervical osteomyelitis is rare, and its occurrence through hematogenous spread in adult horses and the tomographic findings had not been reported previously. The long period of evolution, difficulty in performing an aggressive debridement, and the presence of multi-drug resistant bacteria contributed to the negative outcome.(AU)
Um equino macho, sem raça definida, de 15 anos de idade, foi atendido com histórico de múltiplos abscessos cervicais com evolução de três anos. Na admissão, foram observados: três trajetos fistulosos com drenagem de material purulento na região cervical; baixo escore corporal; restrição de movimentos cervicais; e sensibilidade dolorosa à palpação da região. Foi diagnosticada osteomielite vertebral cervical secundária à infecção por Streptococcus equi. O tratamento inicial consistiu na administração de antibióticos e curativo local. Na ausência de resposta à terapia, realizou-se o debridamento cirúrgico. Inicialmente, obteve-se uma resposta favorável, entretanto, após quatro meses, houve recidiva da lesão e o animal foi submetido à eutanásia. Realizou-se tomografia computadorizada no post mortem para detalhamento da lesão. A osteomielite vertebral cervical é rara, e sua ocorrência por meio de disseminação hematógena em animais adultos não foi previamente reportada. O longo período de evolução, aliado à dificuldade de realização de um debridamento agressivo, e a característica multirresistente do agente etiológico contribuíram para o desfecho negativo do caso.(AU)
Assuntos
Animais , Osteomielite/veterinária , Infecções Estreptocócicas/complicações , Vértebras Cervicais/patologia , Streptococcus equi , CavalosRESUMO
Small children are a challenging group in whom to perform KT. This retrospective study analyzed the results of 62 KTs in children weighing <15 kg, performed between 1998 and 2010, using extraperitoneal access and anastomosis of the renal vessels of donors to the aorta and IVC or iliac vessels of the recipients. Thirty-two (51.6%) grafts were LRDTs and 30 (48.4%) were DDRTs-28 of them pediatric. The mean age at KT was 3.7 ± 2.2 yr (1-12), and the mean weight was 12.3 ± 2.1 kg (5.6-14.9). Ten children weighed <10 kg, and five (8.1%) children presented previous thrombosis of the venous system. At one and five yr, patient survival was 93.2% and 84.2%, and graft survival was 85.2% and 72.7%. There were no differences between the rates for LRDT and DDRT. There were six vascular complications (four vascular thromboses, one laceration, and one renal artery stenosis) and two perirenal collections. Extraperitoneal access is a valid KT technique in children weighing <15 kg.
Assuntos
Peso Corporal , Transplante de Rim/métodos , Anastomose Cirúrgica , Aorta/cirurgia , Criança , Pré-Escolar , Feminino , Taxa de Filtração Glomerular , Sobrevivência de Enxerto , Humanos , Veia Ilíaca/cirurgia , Imunossupressores/uso terapêutico , Lactente , Rim/cirurgia , Masculino , Complicações Pós-Operatórias , Insuficiência Renal , Estudos Retrospectivos , Trombose/patologia , Resultado do Tratamento , Veia Cava Inferior/cirurgiaRESUMO
Trypanosoma cruzi, the causative agent of the Chagas disease, has a complex life cycle alternating between replicative and noninfective forms with nonreplicative and infective forms of the parasite. Metacyclogenesis is a process that takes place in the invertebrate host, comprising morphogenetic transformation from a noninfective form to an infective form, such that parasites acquire the ability to invade human cells. We analyze here the metacyclogenesis process by 2D electrophoresis coupled to MALDI-TOF MS. A large proportion of unique proteins expressed during metacyclogenesis were observed. Interestingly, 50% of the spots were found to differ between epimastigotes and trypomastigotes. We provide a 2D map of the infective metacyclic trypomastigotes. Sixty six protein spots were successfully identified corresponding to 43 different proteins. We analyzed the expression profiles for the identified proteins along metacyclogenesis and classified them into three groups according to their maximal level of expression. We detected several isoforms for a number of proteins, some displaying differential expression during metacyclogenesis. These results suggest that posttranslational modifications may be a fundamental part of the parasite's strategy for regulating gene expression during differentiation. This study contributes to the identification of relevant proteins involved in the metacyclogenesis process. The identification and molecular characterization of these proteins will render vital information about the steps of the parasite differentiation into the infective form.
Assuntos
Proteoma/análise , Proteômica/métodos , Proteínas de Protozoários/análise , Trypanosoma cruzi/metabolismo , Animais , Regulação para Baixo , Eletroforese em Gel Bidimensional , Ponto Isoelétrico , Estágios do Ciclo de Vida , Peso Molecular , Isoformas de Proteínas/análise , Isoformas de Proteínas/metabolismo , Proteoma/metabolismo , Proteínas de Protozoários/metabolismo , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Trypanosoma cruzi/química , Regulação para CimaRESUMO
UNLABELLED: Various strategies have evolved to expand the donor pool due to the extreme shortage of organs. Herein we reviewed our experience with en bloc pediatric kidney transplantation since 1998. METHODS: From January 1998 to December 2004, nine adult patients underwent kidney transplantation using en bloc kidneys from donors <5 years old (range, 1 to 4). The mean age of the recipients was 45.1 years (range, 34 to 57). RESULTS: In recipients of en bloc pediatric transplantation, cold ischemia time ranged from 14 to 26.2 hours (mean, 21.3 hours). Mean serum creatinine at 3, 6, and 12 months after transplantation was 1.53 +/- 0.57, 1.27 +/- 0.27, and 1.15 +/- 0.26 mg/dL compared with 1.93 +/- 1.35, 1.81 +/- 1.17, and 1.73 +/- 0.85 (P = .08) in recipients of single kidneys from ideal cadaveric donors (UNOS criteria, n = 368). Patient and graft survival at 1 year were 88.8% compared with 91.2% and 85% with ideal donors (P = NS), respectively. Three cases required additional surgery. There was one death due to a cerebral vascular accident. CONCLUSION: The present study confirmed the excellent results achieved with transplantation using en bloc kidneys from young donors.
Assuntos
Transplante de Rim/fisiologia , Doadores de Tecidos/estatística & dados numéricos , Adulto , Cadáver , Criança , Pré-Escolar , Creatinina/sangue , Humanos , Lactente , Pessoa de Meia-Idade , Estudos Retrospectivos , Doadores de Tecidos/provisão & distribuiçãoRESUMO
The process of Trypanosoma cruzi metacyclogenesis involves the transformation of noninfective epimastigotes into metacyclic trypomastigotes, which are the pathogenic form. The analysis of stage-specific genes during T. cruzi metacyclogenesis may provide insight into the mechanisms involved in the regulation of gene expression in trypanosomatids. It may also improve the understanding of the mechanisms responsible for the pathology of Chagas disease, and could lead to the identification of new targets for chemotherapy of this disease. We have demonstrated that during metacyclogenesis the expression of several genes is controlled at the translational level by an alternative regulatory mechanism. This mechanism may involve the mobilization of mRNA to the translation machinery. We have been using self-made T. cruzi microarrays to investigate the role of polysomal mobilization in modulating gene expression during metacyclogenesis
Assuntos
Animais , Regulação da Expressão Gênica , Genes de Protozoários , Trypanosoma cruzi/genética , Trypanosoma cruzi/crescimento & desenvolvimento , Estágios do Ciclo de Vida/genética , Trypanosoma cruzi/patogenicidadeRESUMO
The development of the representation of differential expression method has lead to the cloning of Trypanosoma cruzi stage-specific genes. We used this method to characterize a multicopy gene family differentially expressed during metacyclogenesis. The genomic and cDNA clones sequenced encoded three short cysteine-rich polypeptides, of two types, with predicted molecular masses of 7.1, 10.4, and 10.8 kDa. We searched GenBank for similar sequences and found that the sequences of these clones were similar to that encoding the wheat germ agglutinin protein. The region of similarity corresponds to the chitin-binding domain, with eight similarly positioned half-cysteines and conserved aromatic residues involved in chitin recognition. Multiple copies of the genes of this family are present on a high- molecular-mass chromosome. We studied the expression of genes of this family during metacyclogenesis by determining messenger RNA (mRNA) levels. The mRNAs for the members of this gene family were present in the total RNA fraction but were mobilized to the polysomal fraction of adhered (differentiating) epimastigotes during metacyclogenesis, with a peak of accumulation at 24 of differentiation. Polyclonal antisera were raised against a recombinant protein and a synthetic peptide. The specific sera obtained detected 7- and 11-kDa proteins in T. cruzi total protein extracts. The 11-kDa protein was present in similar amounts in the various cell populations, whereas the 7-kDa protein displayed differential synthesis during metacyclogenesis, with maximal levels in 24-h-adhered (differentiating) epimastigotes.
Assuntos
Proteínas de Transporte/genética , Família Multigênica/fisiologia , Proteínas de Protozoários/genética , Trypanosoma cruzi/genética , Sequência de Aminoácidos , Animais , Northern Blotting , Southern Blotting , Western Blotting , Proteínas de Transporte/biossíntese , Proteínas de Transporte/química , Adesão Celular , Quitina/metabolismo , Clonagem Molecular , DNA de Protozoário/química , Expressão Gênica , Genoma de Protozoário , Soros Imunes/química , Dados de Sequência Molecular , Estrutura Terciária de Proteína , Proteínas de Protozoários/biossíntese , Proteínas de Protozoários/química , RNA de Protozoário/análise , Coelhos , Trypanosoma cruzi/química , Trypanosoma cruzi/fisiologiaRESUMO
We isolated a gene that is differentially expressed during Trypanosoma cruzi metacyclogenesis by the representation of differential expression (RDE) method, using differentiating epimastigotes cultured in chemically defined medium. This gene, the metacyclogenin gene, encodes a 630-nucleotide mRNA that is specifically associated with the polysomes of epimastigotes allowed to differentiate for 24 h. We sequenced and characterized the metacyclogenin gene and found that there were at least three copies of the gene organized into tandem 2.8 kb repeats in the genome of T. cruzi Dm28c. We analyzed the repeats and found that they contained two other genes, one encoding tryparedoxin peroxidase and the other encoding a 0.6 kb mRNA (named associated gene or AG) with sequences showing no significant similarity to those in the GenBank database. Northern blot analysis of polysomal RNA extracted from replicating and differentiating epimastigotes showed that metacyclogenin and AG genes displayed similar patterns of expression. Their products were detected only in differentiating epimastigotes, whereas tryparedoxin peroxidase was detected only in the polysomal RNA fraction of replicating and differentiating epimastigotes. In Northern blots of total RNA from differentiating and replicating epimastigotes, the genes studied were detected in both cell populations. The differential expression of the metacyclogenin gene was confirmed by immunocytochemistry studies showing that the protein is detected only in differentiating (adhered) epimastigote. The results suggest that mRNA mobilization to polysomes is an important mechanism in the regulation of gene expression in T. cruzi.
Assuntos
Clonagem Molecular/métodos , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Protozoários/genética , Trypanosoma cruzi/crescimento & desenvolvimento , Trypanosoma cruzi/metabolismo , Sequência de Aminoácidos , Animais , Meios de Cultura , DNA Complementar/genética , Genoma de Protozoário , Estágios do Ciclo de Vida , Dados de Sequência Molecular , Mapeamento Físico do Cromossomo , Proteínas de Protozoários/química , Proteínas de Protozoários/metabolismo , Análise de Sequência de DNA , Trypanosoma cruzi/genéticaRESUMO
The transformation of epimastigotes into metacyclic trypomastigotes involves changes in the pattern of expressed genes, resulting in important morphological and functional differences between these developmental forms of Trypanosoma cruzi. In order to identify and characterize genes involved in triggering the metacyclogenesis process and in conferring to metacyclic trypomastigotes their stage specific biological properties, we have developed a method allowing the isolation of genes specifically expressed when comparing two close related cell populations (representation of differential expression or RDE). The method is based on the PCR amplification of gene sequences selected by hybridizing and subtracting the populations in such a way that after some cycles of hybridization-amplification genes specific to a given population are highly enriched. The use of this method in the analysis of differential gene expression during T. cruzi metacyclogenesis (6 hr and 24 hr of differentiation and metacyclic trypomastigotes) resulted in the isolation of several clones from each time point. Northern blot analysis showed that some genes are transiently expressed (6 hr and 24 hr differentiating cells), while others are present in differentiating cells and in metacyclic trypomastigotes. Nucleotide sequencing of six clones characterized so far showed that they do not display any homology to gene sequences available in the GeneBank.