RESUMO
cDNAs were obtained for macrophage migration-inhibitory factor (MIF)/L-dopachrome methyl ester tautomerase homologues from the parasitic nematodes Trichinella spiralis (TsMIF) and Trichuris trichiura (TtMIF). The translated sequences, which were partly confirmed by sequencing of proteolytic fragments, show 42 and 44% identity respectively with human or mouse MIF, and are shorter by one C-terminal residue. Unlike vertebrate MIF and MIF homologues of filarial nematodes, neither TsMIF nor TtMIF contain cysteine residues. Soluble recombinant TsMIF, expressed in Escherichia coli showed secondary structure (by CD spectroscopy) and quaternary structure (by light-scattering and gel filtration) similar to that of the trimeric mammalian MIFs and D-dopachrome tautomerase. The catalytic specificity of recombinant TsMIF in the ketonization of phenylpyruvate (1.4x10(6) M(-1) x s(-1)) was comparable with that of human MIF, while that of p-hydroxyphenylpyruvate (9.1x10(4) M(-1) x s(-1)) was 71-fold lower. TsMIF showed high specificity in tautomerization of the methyl ester of L-dopachrome compared with non-esterified L-dopachrome (>87000-fold) and a high kcat (approximately 4x10(4) s(-1). The crystal structure, determined to 1.65 A (1 A=0.1 nm), was generally similar to that of human MIF, but differed in the boundaries of the putative active-site pocket, which can explain the low activity towards p-hydroxyphenylpyruvate. The central pore was blocked, but was continuous, with the three putative tautomerase sites. Recombinant TsMIF (5 ng/ml-5 pg/ml) inhibited migration of human peripheral-blood mononuclear cells in a manner similar to that shown by human MIF, but had no effect from 5 to 500 ng/ml on anti-CD3-stimulated murine T-cell proliferation. TsMIF was detected in supernatants of T. spiralis larvae cultured in vitro at 6 ng/ml (55 ng/mg total secreted protein). In conclusion TsMIF has structural, catalytic and cell-migration-inhibitory properties which indicate that it is partially orthologous to mammalian MIF.
Assuntos
Oxirredutases Intramoleculares/química , Fatores Inibidores da Migração de Macrófagos/química , Fatores Inibidores da Migração de Macrófagos/metabolismo , Trichinella spiralis/fisiologia , Trichuris/fisiologia , Sequência de Aminoácidos , Animais , Clonagem Molecular , Cristalografia por Raios X , DNA Complementar , Escherichia coli , Humanos , Oxirredutases Intramoleculares/genética , Oxirredutases Intramoleculares/metabolismo , Cinética , Larva , Fatores Inibidores da Migração de Macrófagos/genética , Camundongos , Modelos Moleculares , Dados de Sequência Molecular , Biossíntese de Proteínas , Estrutura Quaternária de Proteína , Estrutura Secundária de Proteína , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Alinhamento de Sequência , Homologia de Sequência de Aminoácidos , Trichinella spiralis/genética , Trichuris/genética , VertebradosRESUMO
PM10, PM25, precursor gas, and upper-air meteorological measurements were taken in Mexico City, Mexico, from February 23 to March 22, 1997, to understand concentrations and chemical compositions of the city's particulate matter (PM). Average 24-hr PM10 concentrations over the period of study at the core sites in the city were 75 H g/m3. The 24-hr standard of 150 µ g/m3 was exceeded for seven samples taken during the study period; the maximum 24-hr concentration measured was 542 µ g/m3. Nearly half of the PM10 was composed of fugitive dust from roadways, construction, and bare land. About 50% of the PM10 consisted of PM2.5, with higher percentages during the morning hours. Organic and black carbon constituted up to half of the PM2.5. PM concentrations were highest during the early morning and after sunset, when the mixed layers were shallow. Meteorological measurements taken during the field campaign show that on most days air was transported out of the Mexico City basin during the afternoon with little day-to-day carryover.
RESUMO
Recent developments in source apportionment modeling of volatile organic compounds (VOCs) include receptor modeling (RM) applications to "total" (indoor and outdoor) exposure assessment for source of VOC. Source fingerprints are available for major VOC sources such as gasoline vapor, automobile exhaust, refinery emissions, cleaning solvent vapors, printing inks, and waste-water treatment facilities. The relative proportion of each VOC species in the source fingerprint enables the RM method, through a least squares analysis, to identify each source's presence and quantify its contribution to ambient air concentrations. Sampling periods and locations may be selected to represent microenvironmental exposures. Receptor modeling has direct applicability to determining the relative contribution of gasoline vapors to VOC exposures in the general population.