Biomedical subjects

Lin Yan

Publications and source records attributed to Lin Yan.

42 records · Page 3Linked to original sources

PubMed2002

Punctin, a novel ADAMTS-like molecule, ADAMTSL-1, in extracellular matrix.

Punctin (ADAMTSL-1) is a secreted molecule resembling members of the ADAMTS family of proteases. Punctin lacks the pro-metalloprotease and the disintegrin-like domain typical of this family but contains other ADAMTS domains in precise order including four thrombospondin type I repeats. Punctin is the product of a distinct gene on human chromosome 9p21-22 and mouse chromosome 4 that is expressed in adult skeletal muscle. His-tagged punctin expressed in stably transfected High-Five(TM) insect cells was purified to apparent homogeneity by Ni-chromatography of conditioned medium. The NH(2) terminus is not blocked and has the sequence EEDRD and so forth as determined by Edman degradation, demonstrating signal peptidase processing. Recombinant epitope-tagged punctin has a calculated mass of 59,991 Da but exhibits major molecular species of 61970 +/- 6 Da and 62131 +/- 5 Da as measured by liquid chromatography electrospray mass spectrometry. Punctin is a glycoprotein based on carbohydrate staining and liquid chromatography electrospray mass spectrometry glycopeptide analysis. Glycosylation occurs at a single N-linked site as demonstrated by altered electrophoretic migration of punctin expressed in the presence of tunicamycin A. Punctin contains disulfide bonds based on antibody accessibility and electrophoretic migration under reducing versus nonreducing conditions. Rotary shadowing demonstrates that punctin is hatchet-shaped having a globular region attached to a short stem. In transfected COS-1 cells, punctin is deposited in the cell substratum in a punctate fashion and is excluded from focal contacts. Punctin is the first member of a novel family of ADAMTS-like proteins that may have important functions in the extracellular matrix.

ADAMTS Proteins↗

PubMed2002

Dietary supplementation with isolated soy protein reduces metastasis of mammary carcinoma cells in mice.

The present study investigated the effect of dietary supplementation with isolated soy protein (ISP) on pulmonary metastasis of carcinoma cells from primary mammary tumors induced by orthotopic injection of 4526 murine mammary carcinoma cells in female BALB/c mice. Three diets were compared: a basal AIN-93G diet and the basal diet supplemented with 10% or 20% ISP. After three weeks on the experimental diets, each mouse was injected 4 x 10(5) carcinoma cells into the right inguinal mammary fat pad. The primary tumors were excised when they reached a size of 1.0 cm in diameter. After surgery, mice were maintained on their respective diets for another three weeks. At necropsy, the incidence of metastasis, the number and size of macroscopic tumors, and the number of microscopic tumors in the lungs were determined. The incidence of mice with macroscopically visible tumors was 93%, 76%, and 67%, and the median number of macroscopic tumor was 5, 2, and 1 for the control, 10%, and 20% ISP groups (P < or = 0.05, 20% ISP vs. control). The median cross-sectional area of the macroscopic tumors was 0.93 mm2, 0.80 mm2, and 0.31 mm2, and the median volume was 0.73 mm3, 0.56 mm3, and 0.14 mm3 for the control, 10%, and 20% ISP groups (P < or = 0.01, 20% ISP vs. control). Histological examination revealed fewer microscopically detectable tumors in the ISP groups compared with the control. These results demonstrated that dietary supplementation with ISP reduced pulmonary metastasis of carcinoma cells from primary mammary tumors in BALB/c mice.

Animals↗

PubMed2002

Evidence that light modulates protein nitration in rat retina.

As part of ongoing efforts to better understand the role of protein oxidative modifications in retinal pathology, protein nitration in retina has been compared between rats exposed to damaging light or maintained in the dark. In the course of the research, Western methodology for detecting nitrotyrosine-containing proteins has been improved by incorporating chemical reduction of nitrotyrosine to aminotyrosine, allowing specific and nonspecific nitrotyrosine immunoreactivity to be distinguished. A liquid chromatography MS/MS detection strategy was used that selects all possible nitrotyrosine peptides for MS/MS based on knowing the protein identity. Quantitative liquid chromatography MS/MS analyses with tetranitromethane-modified albumin demonstrated the approach capable of identifying sites of tyrosine nitration with detection limits of 4-33 fmol. Using two-dimensional gel electrophoresis, Western detection, and mass spectrometric analyses, several different nitrotyrosine-immunoreactive proteins were identified in light-exposed rat retina compared with those maintained in the dark. Immunocytochemical analyses of retina revealed that rats reared in darkness exhibited more nitrotyrosine immunoreactivity in the photoreceptor outer segments. After intense light exposure, immunoreactivity decreased in the outer segments and increased in the photoreceptor inner segments and retinal pigment epithelium. These results suggest that light modulates retinal protein nitration in vivo and that nitration may participate in the biochemical sequela leading to light-induced photoreceptor cell death. Furthermore, the identification of nitrotyrosine-containing proteins from rats maintained in the dark, under non-pathological conditions, provides the first evidence of a possible role for protein nitration in normal retinal physiology.

Albumins↗

PubMed2002

Cyclic GMP-induced reduction in cardiac myocyte function is partially mediated by activation of the sarcoplasmic reticulum Ca(2+)-ATPase.

We tested the hypothesis that the mechanism through which cyclic GMP reduces cardiac function is mediated by activation of the sarcoplasmic reticulum Ca(2+)-ATPase (SERCA). Cardiac myocytes were isolated from New Zealand white rabbits (n = 11). Individual ventricular cells were stimulated by electrical field stimulation. The maximal rate of cell shortening and percentage shortening were measured with a video edge detector. Thapsigargin (10(-8) mol/l) was used as a specific inhibitor of SERCA. When 8-bromo-cyclic GMP (8-Br-cGMP, 10(-7, -6, -5) mol/l) was added to cells, the maximal rate of myocyte shortening (R(max), microm/s) and percentage shortening were both decreased in a concentration-dependent manner. R(max) decreased 27% from 117 +/- 12 at baseline to 85.2 +/- 13 when 10(-5) mol/l of 8-Br-cGMP was present, and percent shortening was reduced 28% from 6.0 +/- 0.5 to 4.3 +/- 0.5%. Thapsigargin (10(-8) mol/l) increased the maximal rate of myocyte shortening and percent shortening. Addition of thapsigargin prior to 8-Br-cGMP reduced the negative effects of cGMP on myocyte function. The percent shortening decreased only 11% and R(max) decreased 14% with 10(-5) mol/l 8-Br-cGMP, which was not significant. Cyclopiazonic acid, another SERCA inhibitor, was also used to test whether 8-Br-cGMP reduced myocyte function through SERCA. The results were similar to those when thapsigargin was used. These results indicated that the cyclic GMP-induced reduction in cardiac myocyte function was partially mediated through the action of the sarcoplasmic reticulum Ca(2+)-ATPase.

Animals↗

PubMed2002

Genetically engineered crops: their potential use for improvement of human nutrition.

Great success has been achieved in increasing agriculture productivity to fulfill human needs during the second half of the 20th century. However, there will be much greater challenges in the future. Based on the current population growth rate of 1.4% per year, the world's population is forecast to increase from the current level of approximately six billion to nine to twelve billion in 50 years. In addition to continuously increasing demand for agricultural production, there is an urgent need to improve the nutritional quality of human diets for this rapidly growing human population. Malnutrition is still a worldwide health issue. Macronutrient and micronutrient deficiencies are prevalent in developing countries, and over-consumption of certain nutrients in developed countries (e.g., saturated fatty acids) is associated with high incidence of certain chronic diseases. Furthermore, there will be declining natural resources such as arable land and water, and the challenges to humans must be met without further degrading the environment. Biotechnology offers a valuable tool to help achieve these goals. This review focuses on the most recent advances in biotechnology, which promise to improve human nutrition by enhancing the nutrient density of plant foods. Issues relating to the safety of food products from genetically engineered crops are also discussed.

Agriculture↗

PubMed2002

Pigment epithelium-derived factor suppresses ischemia-induced retinal neovascularization and VEGF-induced migration and growth.

PURPOSE: To determine the effect of pigment epithelium-derived factor (PEDF) in a mouse model of ischemia-induced retinal neovascularization and on vascular endothelial growth factor (VEGF)--induced migration and growth of cultured microvascular endothelial cells. METHODS: Human recombinant PEDF was expressed in the human embryonic kidney 293 cell line and purified by ammonium sulfate precipitation and cation exchange chromatography. C57BL/6 mice were exposed to 75% oxygen from postnatal day (P)7 to P12 and then returned to room air. Mice received intravitreal injections of 2 microg PEDF in one eye and vehicle in the contralateral eye on P12 and P14. At P17, mice were killed and eyes enucleated for quantitation of retinal neovascularization. The mitogenic and motogeneic effects of VEGF on cultured bovine retinal and adrenal capillary endothelial cells were examined in the presence or absence of PEDF, using cell counts and migration assays. RESULTS: Two species of human recombinant PEDF, denoted A and B, were purified to apparent homogeneity. PEDF B appeared to comigrate on SDS-PAGE with PEDF from human vitreous samples. Changes in electrophoretic mobility after peptide-N-glycosidase F (PNGase F) digestion suggest that both PEDF forms contain N-linked carbohydrate. Analyses of the intact proteins by liquid chromatography-electrospray mass spectrometry (LC-ESMS) revealed the major molecular weight species for PEDF A (47,705 +/- 4) and B (46,757 +/- 5). LC-ESMS analysis of tryptic peptides indicated that PEDF A and B exhibit differences in glycopeptides containing N-acetylneuraminic acid (NeuAc) and N-acetylhexosamine (HexNAc). Intravitreal administration of either species of PEDF significantly inhibited retinal neovascularization (83% for PEDF A and 55% for PEDF B; P = 0.024 and 0.0026, respectively). PEDF A and B (20 nM) suppressed VEGF-induced retinal microvascular endothelial cell proliferation by 48.8% and 41.4%, respectively, after 5 days (P < 0.001) and VEGF-induced migration by 86.5% +/- 16.7% and 78.1% +/- 22.3%, respectively, after 4 hours (P = 0.004 and P = 0.008, respectively). CONCLUSIONS: These data indicate that elevated concentrations of PEDF inhibit VEGF-induced retinal endothelial cell growth and migration and retinal neovascularization. These findings suggest that localized administration of PEDF may be an effective approach for the treatment of ischemia-induced retinal neovascular disorders.

Animals↗