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J Biochem ; 151(1): 57-64, 2012 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-21893516


Mannan-binding lectin (MBL) was first discovered as a collectin in animal blood, and was shown to have such unique characteristics as a collage-like domain and a carbohydrate recognition domain. We recently identified human collectin kidney 1 (CL-K1, COLEC11) from a human kidney cDNA library. To quantitate the CL-K1 concentration in blood, we developed several polyclonal and monoclonal antibodies using recombinant human CL-K1 in CHO cells and the CL-K1 fragment in Escherichia coli. Using these antibodies, we established a sandwich enzyme-linked immunosorbent assay (ELISA) system. The concentration of CL-K1 in human plasma was 0.34 ± 0.13 µg/ml and that in MBL was 1.72 ± 1.51 µg/ml. Concentrations of MBL are often low due to its single nucleotide polymorphisms (SNPs) which seem to be related to an opsonic defect. However, no low concentrations of CL-K1 were observed on testing over two hundred blood samples. We also found that the blood concentration of CL-K1 was not dependent on gender or age and did not correlate completely with that of MBL. The ELISA system developed in this study will be useful for elucidating the physiological and pathophysiological role of CL-K1 in humans.

Colectinas/sangue , Lectina de Ligação a Manose/sangue , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Anticorpos/imunologia , Anticorpos/metabolismo , Western Blotting , Células CHO , Colectinas/genética , Colectinas/imunologia , Cricetinae , Cricetulus , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Masculino , Lectina de Ligação a Manose/genética , Lectina de Ligação a Manose/imunologia , Pessoa de Meia-Idade , Coelhos , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/metabolismo , Valores de Referência , Adulto Jovem
Biochim Biophys Acta ; 1810(12): 1150-9, 2011 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-22001438


BACKGROUND: Scavenger receptors are generally expressed in macrophages and vascular endothelial cells and some scavenger receptors are thought to contribute to the development of atherosclerosis. METHODS: We cloned the cDNA of a zebrafish CL-P1 (collectin placenta 1) and performed a knockdown study using its antisense morpholino oligonucleotides (MO). RESULTS: Zebrafish CL-P1 (zCL-P1) is 51% identical to human CL-P1 in its amino acid sequence. Microbes and OxLDL bound to zCL-P1 cDNA transfected cells. zCL-P1 mRNA expression gradually increased after 6hours post-fertilization (hpf), reached its highest level at 24hpf, and then decreased, which is similar to the gene expression pattern of Tie-2. The knockdown of zCL-P1 led to an increase in the number of zebrafish embryos with severe morphological abnormalities such as short body lengths and defects in the dorsal aorta at 48hpf. Simultaneous injection of both MO and synthetic zCL-P1 or zVEGF mRNA rescued the abnormal phenotype. CONCLUSIONS: In vivo knockdown study shows that zCL-P1 is implicated in vasculogenesis and those of our in vitro study support its role as a scavenger receptor. GENERAL SIGNIFICANCE: These results suggest that zCL-P1 might be essential for vasculogenesis during the early embryonic phase in bone fish.

Receptores Depuradores/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Células CHO , Clonagem Molecular , Cricetinae , Cricetulus , DNA Complementar , Técnicas de Silenciamento de Genes , Imuno-Histoquímica , Hibridização In Situ , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Receptores Depuradores/fisiologia , Peixe-Zebra
J Nat Med ; 65(1): 103-10, 2011 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-20890669


Genus Lophophora (Cactaceae) has two species: Lophophora williamsii Coulter, which is called peyote, and L. diffusa Bravo. Although it was reported that L. williamsii contained mescaline and L. diffusa did not, we found L. williamsii specimens that did not contain mescaline. This finding indicated that the two species could not be differentiated in terms of mescaline content. Moreover, the relationship between mescaline content and morphology of the two species is also unknown. In this study, we attempted to clarify the difference in morphology, mescaline content, and DNA alignment of the chloroplast trnL/trnF region between L. williamsii and L. diffusa. As a result, L. williamsii specimens were classified into two groups. Group 1 had small protuberances on the epidermis, contained mescaline, and the analyzed region on the trnL/trnF sequence was 881 base pairs (bp) long in all except one (877 bp). Group 2 had large protuberances on the epidermis, did not contain mescaline, and the analyzed region was 893 bp long. On the other hand, L. diffusa had medium-sized protuberances on the epidermis, did not contain mescaline, and the analyzed region was 903 bp long. Also investigated was the potential application of the PCR-restriction fragment length polymorphism (RFLP) method as a means of identification based on the trnL/trnF sequence. By applying the PCR-RFLP method, the two species could be distinguished and L. williamsii specimens could be differentiated into group 1 and group 2.

Cactaceae/química , Cactaceae/genética , Mescalina/química , Cactaceae/ultraestrutura , Microscopia Eletrônica de Varredura , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição
J AOAC Int ; 94(6): 1770-7, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-22320083


An analog of aildenafil, which is a potent and highly selective inhibitor of phosphodiesterase 5, was found in a dietary supplement marketed for enhancement of sexual function. The compound was isolated by silica gel column chromatography, and its structure was identified by means of 13C-NMR spectrometry, 1H-NMR spectrometry, high-resolution MS, and X-ray structure determination. The compound was identified to be sulfoaildenafil (other names: thioaildenafil, dimethyl sildenafil thione, and thiomethisosildenafil). Sulfoaildenafil is very similar to the compound thiohomosildenafil. As it is difficult to distinguish between them by LC-photodiode array detector analysis, ultra-performance LC (UPLC)/MS, ion trap LC/MS/MS (LC/IT-MS/MS), and GC/MS were performed. The mass spectra of thiohomosildenafil by UPLC/MS and LC/IT-MS/MS showed mass fragments of m/z 58, 72, and 355, and the mass spectrum by GC/MS showed mass fragments of m/z 56, 72, and 420. Some of these fragments had low intensities, but they were useful for distinguishing between the two compounds. The relationship between aildenafil (other names: dimethylsildenafil and methisosildenafil) and homosildenafil is similar to that between sulfoaildenafil and thiohomosildenafil. Therefore, these compounds were also examined.

Suplementos Nutricionais/análise , Piperazinas/análise , Sulfonas/análise , Cromatografia Líquida/métodos , Nucleotídeo Cíclico Fosfodiesterase do Tipo 5/análise , Cromatografia Gasosa-Espectrometria de Massas/métodos , Espectroscopia de Ressonância Magnética/métodos , Estrutura Molecular , Inibidores da Fosfodiesterase 5/análise , Espectrometria de Massas em Tandem/métodos
Dalton Trans ; (3): 473-7, 2006 Jan 21.
Artigo em Inglês | MEDLINE | ID: mdl-16395447


We have designed and synthesised a new carborane derivative containing two nitronyl nitroxides. This molecule can be considered as a prototype of a three-dimensional analogue of o-benzoquinodimethane. The magnetic susceptibility of a crystalline sample revealed that this biradical possesses a weak antiferromagnetic interaction (theta = -1.05(4) K) and an ESR study in frozen matrix in the 5.4-104 K range gave an intramolecular antiferromagnetic interaction of -27(2) K. The synthesis, physical properties and DFT calculation result are also reported.