Epidemiología Inversa: baja concentración del LDL modificadas y morbimortalidad cardiovascular en pacientes en hemodiálisis / Reverse epidemiology: low concentration of modified LDL and cardiovascular morbidity and mortality in hemodialysis patients / Epidemiologia Reversa: baixa concentração de LDL modificadas e morbimortalidade cardiovascular em pacientes em hemodiálise

La disminución de colesterol-LDL (c-LDL) se considera meta principal del tratamiento de pacientes con riesgo cardiovascular. Sin embargo, pacientes con Enfermedad Renal Crónica (ERC) en hemodiálisis presentan c-LDL menor de 100 mg/dL, aumentos moderados de triglicéridos y baja frecuencia de colesterol-HDL por debajo de valores deseables. Esta condición se encuadra dentro del fenómeno conocido como "epidemiología inversa", en la cual la conocida asociación prevalente entre hipercolesterolemia, hipertensión arterial, obesidad y morbimortalidad por eventos cardiovasculares no se encuentra y, por el contrario se invierte la estrecha relación de estos parámetros con eventos cardiovasculares propia de los pacientes no hemodializados. Por un lado el 35% de los pacientes con ERC presentan diabetes mellitus tipo 2 y por otra parte, existen otros factores patogénicos menos conocidos como la Lipoproteína asociada a Fosfolipasa A2, la Proteína C Reactiva, los remanentes lipoproteicos, la Lp(a) y enzimas y proteínas asociadas a la HDL, como la Paraoxonasa y Apo A-I. El conjunto de factores descritos podrían reemplazar, en pacientes con ERC en hemodiálisis, al colesterol-LDL (c-LDL), típico analito que en otros pacientes actúa como factor de riesgo y/o patogénico de aterosclerosis y no sólo como marcador circulante. Una explicación plausible respecto al c-LDL disminuído es la modificación cualitativa de LDL por oxidación, glicación, carbamilación, la presencia de LDL pequeñas y densas, fenómenos inflamatorios y malnutrición.

Proteomic Analysis of Colonic Mucosal Tissue from Tuberculous and Ulcerative Colitis Patients

Changes in the expression profiles of specific proteins leads to serious human diseases, including colitis. The proteomic changes related to colitis and the differential expression between tuberculous (TC) and ulcerative colitis (UC) in colon tissue from colitis patients has not been defined. We therefore performed a proteomic analysis of human TC and UC mucosal tissue. Total protein was obtained from the colon mucosal tissue of normal, TC, and UC patients, and resolved by 2-dimensional electrophoresis (2-DE). The results were analyzed with PDQuest using silver staining. We used matrix-assisted laser desorption ionization time-of-flight/time-of-flight spectrometry (MALDI TOF/TOF) to identify proteins differentially expressed in TC and UC. Of the over 1,000 proteins isolated, three in TC tissue and two in UC tissue displayed altered expression when compared to normal tissue. Moreover, two proteins were differentially expressed in a comparative analysis between TC and UC. These were identified as mutant beta-actin, alpha-enolase and Charcot-Leyden crystal protein. In particular, the expression of alpha-enolase was significantly greater in TC compared with normal tissue, but decreased in comparison to UC, implying that alpha-enolase may represent a biomarker for differential diagnosis of TC and UC. This study therefore provides a valuable resource for the molecular and diagnostic analysis of human colitis.

[Molecular detection of phospholipase B3 as an effective factor in virulence of Aspergillus fumigatus]

Phospholipases are the major component of all eukaryotic bilayer cell membranes. and serve as a scaffold for membranes proteins. Phospholipases are a class of ubiquitous enzymes that have estrase activity and due to their ability to lyses of cell membrane can play an important role for pathogenicity of microrganisims. During this study using degenerate primers based on homologous amino acid sequences of some fungi phospholipase B [PLB], detection of Aspergillus fumigatus PLB3 was carried out. After DNA extraction of A. fumigatus, A. niger, Fusarium venonatum and Aurobasidium pullulans PCR reaction was carried out using degenerate primers. Predicted 2100 bp product from A. fumigatus was cloned in pGEMT-Easy vector and then transformed into E. coli Top 10 F' competent cell for extraction of cloned DNA fragment. Sequence analysis of 2100 bp fragments using BLASTX software were revealed a high homology to published PLB sequences from other fungi. Phylogenic tree analysis of PLB3 gene shows that PLB and potential PLB analogues like lysophospholipase are contained in a large cluster of the PLB family. A. fumigatus PLB gene is more closely related to A. oryzae, A. niger, Penicillium chrysogenum and Neurospora crassa than other fungi. Also, sequence availability of full length PLB3 gene represents a noteworthy breakthrough in the study of this opportunistic pathogen and function of PLB3 gene in pathogenesis of A. fumigates. Detection of PLB3 gene can be lead to preparing vaccine, blocker and laboratory marker for recognition of PLB3 gene products either in patient body or clinical samples

Cloning and sequencing of partial plb2 gene of Aspergillus fumigatus

Aspergillus species are primarily opportunistic pathogens and with a few exceptions cause invasive disease only in an immunocompromised hosts during last two decades, the importance of fungal disease has been increased dramatically due to an increased occurrence of tuberculosis, chemotherapy-induced neutropenia, AIDS, open chest surgery and use of antibiotics and immunosuppressive therapy. The production of extracellular phospholipases by pathogenic fungi such as A. fumigatus are involved in the degradation of target cell membrane phospholipids in invaded tissues. Using degenerate primers based on homologous amino acid sequences of fungal phospholipase B [PLB] gene, PCR products of 542 was generated which was the predicted size. Sequence analysis revealed sequence had a high homology to published PLB sequences sharing highest homology to Aspergillus oryzae PLB [62% identity]. Inverse PCR was used to attempt to clone the entire plb2 genes. Initially, genomic DNA was digested with selected restriction enzymes and self ligated with T4 DNA ligase to give circular DNA for inverse PCR [IPCR] for A. fumigatus plb2. IPCR with Xho I self-ligated digest gave product of 1.2 Kb for plb2 and was found to encode the 3' ends of gene and using CAP assembly software this sequence was assembled with 542 bp sequence. BLST X and Gene Finder analysis revealed that the first 542 bp of assembled total 1529 bp [after deletion of similar bp from both sequences] encoded the 3' end of plb2 gene. The sequence availability of partial afplb2 gene and after that full sequence of this gene represents a major breakthrough in the study of this opportunistic pathogen and function of plb gene in pathogenesis of A. fumigatus. Also phylogenic tree analysis of afplb2 gene shows that PLBs and potential PLB analogues are contained in a large cluster of the PLB family. A. fumigatus are more closely relaed to A. oryzae, A. niger, P. notatum and N. crassa PLB than other fungi

A Case of Bronchial Obstruction Caused by Charcot-Leyden Crystals / 소아알레르기및호흡기학회지

Charcot-Leyden crystals are slender, hexagonal, bipyramidal crystals formed predominantly from eosinophil but also from basophil granules composed of a single protein of 17,400 daltons of lysophospholipase activity. It has been identified in a variety of body fluids and tissues in human tissues and secretions in association with eosinophilic inflammatory reactions, asthma, myeloid leukemias, and allergic, parasitic, and other diseases. In this report, we describe a child with bronchial obstruction and subsequent atelectasis caused by Charcot-Leyden crystals containing fibronous material who was treated with flexible bronchofiberscope to remove it.