Proteomic profile of culture filtrate from the Brazilian vaccine strain Mycobacterium bovis BCG Moreau compared to M. bovis BCG Pasteur.

BACKGROUND: Bacille Calmette-Guerin (BCG) is currently the only available vaccine against tuberculosis (TB) and comprises a heterogeneous family of sub-strains with genotypic and phenotypic differences. The World Health Organization (WHO) affirms that the characterization of BCG sub-strains, both on genomic and proteomic levels, is crucial for a better comprehension of the vaccine. In addition, these studies can contribute in the development of a more efficient vaccine against TB. Here, we combine two-dimensional electrophoresis (2DE) and mass spectrometry to analyse the proteomic profile of culture filtrate proteins (CFPs) from M. bovis BCG Moreau, the Brazilian vaccine strain, comparing it to that of BCG Pasteur. CFPs are considered of great importance given their dominant immunogenicity and role in pathogenesis, being available for interaction with host cells since early infection. RESULTS: The 2DE proteomic map of M. bovis BCG Moreau CFPs in the pH range 3-8 allowed the identification of 158 spots corresponding to 101 different proteins, identified by MS/MS. Comparison to BCG Pasteur highlights the great similarity between these BCG strains. However, quantitative analysis shows a higher expression of immunogenic proteins such as Rv1860 (BCG1896, Apa), Rv1926c (BCG1965c, Mpb63) and Rv1886c (BCG1923c, Ag85B) in BCG Moreau when compared to BCG Pasteur, while some heat shock proteins, such as Rv0440 (BCG0479, GroEL2) and Rv0350 (BCG0389, DnaK), show the opposite pattern. CONCLUSIONS: Here we report the detailed 2DE profile of CFPs from M. bovis BCG Moreau and its comparison to BCG Pasteur, identifying differences that may provide relevant information on vaccine efficacy. These findings contribute to the detailed characterization of the Brazilian vaccine strain against TB, revealing aspects that may lead to a better understanding of the factors leading to BCG's variable protective efficacy against TB.

Characterization of the Opp peptide transporter of Corynebacterium pseudotuberculosis and its role in virulence and pathogenicity.

Despite the economic importance of caseous lymphadenitis (CLA), a chronic disease caused by Corynebacterium pseudotuberculosis, few genes related to the virulence of its etiologic agent have been characterized. The oligopeptide permease (Opp) transporters are located in the plasma membrane and have functions generally related to the uptake of peptides from the extracellular environment. These peptide transporters, in addition to having an important role in cell nutrition, also participate in the regulation of various processes involving intercellular signaling, including the control of the expression of virulence genes in pathogenic bacteria. To study the role of Opp in C. pseudotuberculosis, an OppD deficient strain was constructed via simple crossover with a nonreplicative plasmid carrying part of the oppD gene sequence. As occurred to the wild-type, the ΔoppD strain showed impaired growth when exposed to the toxic glutathione peptide (GSH), indicating two possible scenarios: (i) that this component can be internalized by the bacterium through an Opp-independent pathway or (ii) that there is toxicity while the peptide is extracellular. Additionally, the ΔoppD mutant presented a reduced ability to adhere to and infect macrophages compared to the wild-type, although both strains exhibit the same potential to colonize spleens and cause injury and death to infected mice.

Genome Sequence of Lactococcus lactis subsp. lactis NCDO 2118, a GABA-Producing Strain.

Lactococcus lactis subsp. lactis NCDO 2118 is a nondairy lactic acid bacterium, a xylose fermenter, and a gamma-aminobutyric acid (GABA) producer isolated from frozen peas. Here, we report the complete genome sequence of L. lactis NCDO 2118, a strain with probiotic potential activity.

Motor function measure scale, steroid therapy and patients with Duchenne muscular dystrophy.

OBJECTIVE: To assess the evolution of motor function in patients with Duchenne muscular dystrophy (DMD) treated with steroids (prednisolone or deflazacort) through the Motor Function Measure (MFM), which evaluates three dimensions of motor performance (D1, D2, D3). METHODS: Thirty-three patients with DMD (22 ambulant, 6 non-ambulant and 5 who lost the capacity to walk during the period of the study) were assessed using the MFM scale six times over a period of 18 months. RESULTS: All the motor functions remained stable for 14 months in all patients, except D1 for those who lost their walking ability. In ambulant patients, D2 (axial and proximal motor capacities) motor functions improved during six months; an improvement in D3 (distal motor capacity) was noted during the total follow-up. D1 (standing posture and transfers) and total score were useful to predict the loss of the ability to walk. CONCLUSIONS: The use of the MFM in DMD patients confirms the benefits of the steroid treatment for slowing the progression of the disease.

Motor function measure scale, steroid therapy and patients with Duchenne muscular dystrophy / Medida de função motora, corticoterapia e pacientes com distrofia muscular de Duchenne

OBJECTIVE: To assess the evolution of motor function in patients with Duchenne muscular dystrophy (DMD) treated with steroids (prednisolone or deflazacort) through the Motor Function Measure (MFM), which evaluates three dimensions of motor performance (D1, D2, D3). METHODS: Thirty-three patients with DMD (22 ambulant, 6 non-ambulant and 5 who lost the capacity to walk during the period of the study) were assessed using the MFM scale six times over a period of 18 months. RESULTS: All the motor functions remained stable for 14 months in all patients, except D1 for those who lost their walking ability. In ambulant patients, D2 (axial and proximal motor capacities) motor functions improved during six months; an improvement in D3 (distal motor capacity) was noted during the total follow-up. D1 (standing posture and transfers) and total score were useful to predict the loss of the ability to walk. CONCLUSIONS: The use of the MFM in DMD patients confirms the benefits of the steroid treatment for slowing the progression of the disease.

OBJETIVO: Avaliar a evolução da função motora de pacientes com distrofia muscular de Duchenne (DMD) em corticoterapia (predinisolona e deflazacort), por meio da escala Medida da Função Motora (MFM), que avalia três dimensões de funções motoras (D1, D2, D3). MÉTODOS: Trinta e três pacientes com DMD (22 deambulantes, seis cadeirantes e cinco que perderam a capacidade de andar ao longo do estudo) foram avaliados pela escala MFM em seis momentos durante 18 meses. RESULTADOS: Todas as funções motoras mantiveram-se estáveis durante 14 meses, exceto D1 para os pacientes que perderam a marcha. Nos pacientes deambulantes, a D2 (função motora axial e proximal) apresentou melhora durante seis meses. Melhora em D3 (função motora distal) também foi observada durante o seguimento. A D1 (postura em pé e transferências) e o escore total foram importantes para predizer a perda de marcha. CONCLUSÕES: O uso da MFM nos pacientes com DMD confirma os benefícios do tratamento com corticoides na diminuição da velocidade de progressão da doença.

Abscesso Hepático por Osso de Peixe / Liver Abscess Caused by Fish Bone

O abscesso hepático originado por corpo estranho é raro e o quadro clínico apresentado inespecífico. O caso clínico relatado demonstra a evolução de um paciente com quadro de dor abdominal no hipogástrio, irradiada para o hipocôndrio direito e associada à febre diária (39ºC). Ao exame físico foi encontrado abdome dolorosoà palpação no hipocôndrio direito. Exames complementares de entrada revelaram leucocitose, AST, ALT,bilirrubinas e alfa-feto proteína dentro da normalidade. Ao ultra-som, massa compatível com abscesso hepá-tico e tomografia computadorizada (TC) com imagem nodular mal definida, localizada em segmento lateral do lobo hepático esquerdo.O tratamento combinou drenagem percutânea guiada por TC e antibioticoterapia - Ampicilina, Garamicina eMetronidazol. Posteriormente à hemocultura (positiva para S. pneumoniae), a Ampicilina foi substituída por Penicilina G cristalina. Por má evolução do caso, optou-se por laparotomia exploratória, que evidenciou perfuração da região pré-pilórica, bloqueada pelo lobo hepático esquerdo e presença de corpo estranho (osso de peixe). Assim, fez-se uma nova drenagem do abscesso com necrosectomia cirúrgica, rafia simples da perfuração gástrica e mantidos os antibióticos. Atualmente, o paciente encontra-se assintomático.Apesar da TC e da endoscopia digestiva alta (EDA), o diagnóstico foi feito apenas no intra-operatório. Diante da alta mortalidade (8,5) apresentada pelos casos de abscesso hepático, é evidente a relação do abscesso hepático no lobo esquerdo com a presença de corpo estranho.

Liver abscess caused by foreign body is rare and the clinical picture presented is non-specific. The reportedclinical case shows a clinical picture of a patient’s evolution with abdominal pain in the epigastric regionradiating towards the right hypochondrium and associated to daily fever (39°C). On physical examination, itwas found a painful abdomen on palpation in the right hypochondrio. Complementary exams of leucocytosis,aspartate aminotransferase, alanine aminotransferase, bilirubin, and alpha-fetus protein on admission wereconsidered normal. Ultrasonography revealed a compatible mass with hepatic abscess, and the computerizedtomography (CT) showed a poorly defined nodular image located in the left lateral division of left lobe of liver. The treatment combined percutaneous drainage guided by CT and antibioticotherapy - Ampicillin, Garamycinand Metronidazol. After the blood culture (positive for S. pneumoniae), Ampicillin was replaced by Penicillincrystalline G. Because of poor evolution of the case, it was decided to perform an exploratory laparotomy thathighlighted the perforation of the prepyloric region blocked by the left lobe of liver and the presence of aforeign body (fish bone). Thus, a new drainage of the abscess with surgical débridement and a simple surgicalsuture of the gastric perforation were carried out. The antibiotics were continued. Nowadays, the patientbecame asymptomatic. In spite of CT and of high digestive endoscopy (HDE), the diagnosis was only performed intraoperatively.Due to high mortality (8.5%) presented by the cases of liver abscess, the relation of a liver abscess in the leftlobe of liver with the presence of a foreign body is evident.