Effect of One Year of Cryopreservation on the Activity of Lysosomal Hydrolases from EBV-Transformed Lymphocytes

Background. The Epstein-Barr virus (EBV) was used as an agent of B lymphocyte proliferation for subsequent diagnosis of lysosomal storage disease. Due to the constant handling of long-preserved samples in our cell bank, we decided to observe the behavior and then compare cultured and frozen samples for at least one year's cryopreservation. Methods. Twenty-five samples from healthy individuals were used to assess the possible changes in activity of enzymes β-galactosidase, β-glucosidase, α-iduronidase, α-galactosidase, and α-glucosidase. Transmission electron microscopy was used to confirm cell transformation of B lymphocytes into EBV-infected cells, generating lymphoblastoid cell lines. Results. Transmission electron microscopy findings confirmed previous reports in the literature that is, significant and evident morphological changes in the nucleus occur after day 12 and the consequent cell transformation into EBV-infected cells. After thawing and subsequent treatment with the five enzymes utilized, we observed no significant changes in samples cryopreserved for more than one year, as compared to samples cultured for 12 days

Diagnosing mucopolysaccharidosis IVA

Mucopolysaccharidosis IVA (MPS IVA; Morquio A syndrome) is an autosomal recessive lysosomal storage disorder resulting from a deficiency of N-acetylgalactosamine-6-sulfate sulfatase (GALNS) activity. Diagnosis can be challenging and requires agreement of clinical, radiographic, and laboratory findings. A group of biochemical genetics laboratory directors and clinicians involved in the diagnosis of MPS IVA, convened by BioMarin Pharmaceutical Inc., met to develop recommendations for diagnosis. The following conclusions were reached. Due to the wide variation and subtleties of radiographic findings, imaging of multiple body regions is recommended. Urinary glycosaminoglycan analysis is particularly problematic for MPS IVA and it is strongly recommended to proceed to enzyme activity testing even if urine appears normal when there is clinical suspicion of MPS IVA. Enzyme activity testing of GALNS is essential in diagnosing MPS IVA. Additional analyses to confirm sample integrity and rule out MPS IVB, multiple sulfatase deficiency, and mucolipidoses types II/III are critical as part of enzyme activity testing. Leukocytes or cultured dermal fibroblasts are strongly recommended for enzyme activity testing to confirm screening results. Molecular testing may also be used to confirm the diagnosis in many patients. However, two known or probable causative mutations may not be identified in all cases of MPS IVA. A diagnostic testing algorithm is presented which attempts to streamline this complex testing process

Astyanax vermilion and Astyanax burgerai: new characid fishes (Ostariophysi: Characiformes) from Northeastern Bahia, Brazil

Two new Astyanax species, A. vermilion, from rio Almada and rio Cachoeira, and A. burgerai, from rio Almada, Bahia State, Brazil, are described. Astyanax vermilion is distinguished from most of its congeners and from all other Astyanax species known from northeastern Brazilian drainages by having distal portion of pelvic fins dark, a combination of sexually dimorphic characters, posteroventral portion of body and fins (except pectoral) reddish in life, and inconspicuous humeral and caudal spots. It also differs by having highest body depth just anterior of dorsal-fin origin, 32-34 lateral-line scales, and presence of one or two maxillary teeth. Astyanax burgerai is diagnosed by the presence of two vertically elongated humeral blotches, absence of a conspicuous and broad dark midlateral stripe (at least on anterior half of body), body highest along vertical slightly behind midlength of pectoral fin, 31-34 lateral-line scales, and two or three teeth on premaxillary outer series

Influência da massa magra corporal nas repercussões cardiopulmonares durante o teste de caminhada de seis minutos em pacientes com DPOC Influence of lean body mass on cardiopulmonary repercussions during the six-minute walk test in patients with COPD

OBJETIVO: A massa magra corporal (MMC) tem sido associada à mortalidade em pacientes com DPOC, mas seu impacto na limitação funcional é pouco conhecido. O objetivo deste trabalho foi analisar as variáveis cardiopulmonares em pacientes com DPOC, com ou sem depleção da MMC, antes e após a realização do teste de caminhada de seis minutos (TC6). MÉTODOS: Foram avaliados pacientes com DPOC, 36 sem depleção de MMC e 32 com depleção de MMC. Todos os pacientes foram submetidos à avaliação clínica, espirometria, avaliação da composição da massa corpórea e TC6 e responderam a questionários de qualidade de vida e de percepção de dispnéia. RESULTADOS: Não foram observadas diferenças significativas na gravidade de obstrução das vias aéreas, na percepção da dispnéia e na qualidade de vida entre os grupos. A distância percorrida no TC6 foi similar nos pacientes com DPOC com e sem depleção de MMC (470,3 ± 68,5 m vs. 448,2 ± 89,2 m). Entretanto, durante a realização do teste, os pacientes com depleção de MMC apresentaram aumento significativamente maior na diferença entre os valores final e basal da frequência cardíaca e do índice da escala de Borg para cansaço dos membros inferiores. A distância percorrida no TC6 apresentou correlação significativa positiva com o VEF1 (r = 0,381; p = 0,01). CONCLUSÕES: Não houve influência da depleção da MMC na capacidade funcional de exercício e na qualidade de vida dos pacientes estudados. Entretanto, os pacientes com depleção de MMC apresentam sintomas de fadiga dos membros inferiores mais acentuados durante o TC6, o que reforça a importância da avaliação e tratamento das manifestações sistêmicas da DPOC.<br>OBJECTIVE: Although lean body mass (LBM) has been associated with mortality in patients with COPD, its influence on functional limitation is not clear. The objective of this study was to analyze the cardiopulmonary variables in COPD patients with or without LBM depletion, prior to and after the six-minute walk test (6MWT). METHODS: We evaluated COPD patients, 32 with LBM depletion and 36 without. All patients underwent clinical evaluation, spirometry, evaluation of body mass composition and 6MWT, as well as completing questionnaires related to quality of life and perception of dyspnea. RESULTS: No significant differences in the severity of airway obstruction, perception of dyspnea and quality of life scores were found between the groups. The distance covered on the 6MWT was similar in COPD patients with and without LBM depletion (470.3 ± 68.5 m vs. 448.2 ± 89.2 m). However, patients with LBM depletion presented significantly greater differences between baseline and final values in terms of heart rate and Borg scale index for lower limb fatigue. There was a significant positive correlation between distance covered on the 6MWT and FEV1 (r = 0.381, p = 0.01). CONCLUSIONS: In the patients studied, functional exercise tolerance and quality of life were unaffected by LBM depletion. However, the patients with LBM depletion presented more pronounced lower limb fatigue during the 6MWT, which underscores the importance of the evaluation and treatment of systemic manifestations in COPD patients