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Biomedical subjects

Hai Yan

Publications and source records attributed to Hai Yan.

9 recordsLinked to original sources
PubMed2003

Transforming single DNA molecules into fluorescent magnetic particles for detection and enumeration of genetic variations.

Many areas of biomedical research depend on the analysis of uncommon variations in individual genes or transcripts. Here we describe a method that can quantify such variation at a scale and ease heretofore unattainable. Each DNA molecule in a collection of such molecules is converted into a single magnetic particle to which thousands of copies of DNA identical in sequence to the original are bound. This population of beads then corresponds to a one-to-one representation of the starting DNA molecules. Variation within the original population of DNA molecules can then be simply assessed by counting fluorescently labeled particles via flow cytometry. This approach is called BEAMing on the basis of four of its principal components (beads, emulsion, amplification, and magnetics). Millions of individual DNA molecules can be assessed in this fashion with standard laboratory equipment. Moreover, specific variants can be isolated by flow sorting and used for further experimentation. BEAMing can be used for the identification and quantification of rare mutations as well as to study variations in gene sequences or transcripts in specific populations or tissues.

Base Sequence↗
PubMed2003

[Toxic effects of hexavalent chromium on the growth of blue-green microalgae].

A standard method of algal bioassay for determining the toxicity of toxic chemicals was applied to the interaction between Cr(VI) and six blue-green microalgae. The 96h-EC50 of hexavalent chromium on the growth of Chlorella pyrenoidosa, Synechococcus, Spirulina maxima, Spirulina platensis, Selenastrum capriornutum and Scenedesmus quadricauda were 4.96, 6.50, 11.16, 11.74, 12.43 and 20.89 mg/L, respectively. The tolerance of the six blue-green microalgae to Cr(VI) was in the order of Scenedesmus quadricauda > Selenastrum capriornutum > Spirulina platensis > Spirulina maxima > Synechococcus > Chlorella pyrenoidosa. The sensitivity of these species to Cr(VI) might relate to the cell size, the structure of cell wall, as well as the redox reaction caused by the exudate or enzyme. X-ray absorption near edge structure(XANES) was used to study the accumulation and transformation of Cr(VI) in Chlorella pyrenoidosa.

Carcinogens, Environmental↗
PubMed2002

VGF is required for obesity induced by diet, gold thioglucose treatment, and agouti and is differentially regulated in pro-opiomelanocortin- and neuropeptide Y-containing arcuate neurons in response to fasting.

Targeted deletion of the gene encoding the neuronal and neuroendocrine secreted polypeptide VGF (nonacronymic) produces a lean, hypermetabolic mouse. Consistent with this phenotype, VGF mRNA levels are regulated in the hypothalamic arcuate nucleus in response to fasting. To gain insight into the site(s) and mechanism(s) of action of VGF, we further characterized VGF expression in the hypothalamus. Double-label studies indicated that VGF and pro-opiomelanocortin were coexpressed in lateral arcuate neurons in the fed state, and that VGF expression was induced after fasting in medial arcuate neurons that synthesize neuropeptide Y (NPY). Like NPY, VGF mRNA induction in this region of the hypothalamus in fasted mice was inhibited by exogenous leptin. In leptin-deficient ob/ob and receptor-mutant db/db mice, VGF mRNA levels in the medial arcuate were elevated. To identify neural pathways that are functionally compromised by Vgf ablation, VGF mutant mice were crossed with obese A(y)/a (agouti) and ob/ob mice. VGF deficiency completely blocked the development of obesity in A(y)/a mice, whereas deletion of Vgf in ob/ob mice attenuated weight gain but had no impact on adiposity. Hypothalamic levels of NPY and agouti-related polypeptide mRNAs in both double-mutant lines were dramatically elevated 10- to 15-fold above those of wild-type mice. VGF-deficient mice were also found to resist diet- and gold thioglucose-induced obesity. These data and the susceptibility of VGF mutant mice to monosodium glutamate-induced obesity are consistent with a role for VGF in outflow pathways, downstream of hypothalamic and/or brainstem melanocortin 4 receptors, that project via the autonomic nervous system to peripheral metabolic tissues and regulate energy homeostasis.

Agouti Signaling Protein↗
PubMed2002

Allele separation facilitates interpretation of potential splicing alterations and genomic rearrangements.

Mutations that alter normal splice patterns and genomic rearrangements are common causes of hereditary diseases including hereditary nonpolyposis colorectal cancer. However, abnormal transcripts can be difficult to detect and interpret because splicing patterns are often heterogeneous even in normal cells. Standard techniques including sequencing and Southern hybridization fail to detect some genomic rearrangements. We show here that separation of alleles in somatic cell hybrids, through "conversion" technology, considerably facilitates the interpretation of abnormal splicing patterns and the detection of genomic rearrangements. We detected novel mutations in MLH1 in each of four hereditary nonpolyposis colorectal cancer patients. The genomic mutations were CAG>CAA predicting Q346Q; GAG>AAG predicting E102K; a>g at nucleotide 1559-2 at intron 13, and a tandem duplication involving exons 7-12. By separating the two alleles, we showed that one allele produced only abnormal transcript or no transcript whereas the other allele produced only normal transcript. These results allowed pathogenicity to be unambiguously assigned to the mutations and increased the sensitivity of genomic testing.

Adaptor Proteins, Signal Transducing↗
PubMed2002

Stat2 binding to the interferon-alpha receptor 2 subunit is not required for interferon-alpha signaling.

The interferon-alpha (IFNalpha) receptor consists of two subunits, the IFNalpha receptor 1 (IFNaR1) and 2 (IFNaR2) chains. Following ligand binding, IFNaR1 is phosphorylated on tyrosine 466, and this site recruits Stat2 via its SH2 domain. In contrast, IFNaR2 binds Stat2 constitutively. In this study we have characterized the Stat2-IFNaR2 interaction and examined its role in IFNalpha signaling. Stat2 binds the major IFNaR2 protein but not a variant containing a shorter cytoplasmic domain. The interaction does not require a STAT SH2 domain. Both tyrosine-phosphorylated and non-phosphorylated Stat2 bind IFNaR2 in vitro; however, relatively little phosphorylated Stat2 associates with IFNaR2 in vivo. In vitro binding assays defined IFNaR2 residues 418-444 as the minimal interaction domain and site-specific mutation of conserved acidic residues within this domain disrupted in vitro and in vivo binding. An IFNaR2 construct carrying these mutations was either (i) overexpressed in 293T cells or (ii) used to complement IFNaR2-deficient U5A cells. Unexpectedly, the activity of an IFNalpha-dependent reporter gene was not reduced but, instead, was enhanced up to 2-fold. This suggests that this particular IFNaR2-Stat2 interaction is not required for IFNalpha signaling, but might act to negatively inhibit signaling. Finally, a doubly truncated recombinant fragment of Stat2, spanning residues 136-702, associated with IFNaR2 in vitro, indicating that the interaction with IFNaR2 is direct and occurs in a central region of Stat2 marked by a hydrophobic core.

Amino Acid Sequence↗
PubMed2002

Toxicity and bioaccumulation of copper in three green microalgal species.

The effective concentrations of copper on the inhibition of the growth of Scenedesmus obliquus, Chlorella pyrenoidosa and Closterium lunula at 96 h (96 h EC50) were determined to be 50, 68 and 200 microg/l, respectively. The low initial bioaccumulation of Cu by C. lunula was found to be responsible for its tolerance to Cu. The amount of Cu accumulated by all three microalgae reached the maximum value and decreased quickly after the peak followed by a slow decrease over the next 6 d. Bioaccumulation of Cu by C. lunula was directly proportional to the initial Cu concentration. After reaching the first peak after 1 d, the bioconcentration factor of Cu by microalgae declined to its minimum value during the exponential growth phase but increased in the stationary growth phase again. This indicates that desorption of Cu from microalgae was higher during the exponential growth phase but lower in the stationary growth phase. Smaller microalgae with low 96 h EC50 values are more efficient in removing Cu from wastewater.

Chlorella↗
PubMed2002

Effect and mechanism of inorganic carbon on the biodegradation of dimethyl phthalate by Chlorella pyrenoidosa.

The effect and mechanism of inorganic carbon (IC) on the biodegradation of dimethyl phthalate (DMP) by a green microalga Chlorella pyrenoidosa was investigated. It was indicated that DMP could be used as the sole carbon source to support the slow heterotrophic growth of C. pyrenoidosa, but both the growth of C. pyrenoidosa and the biodegradation rate of DMP were obviously increased when initial inorganic carbon concentration (IC) was increased from 0.6 to 23.7 mg/l. Phthalic acid (PA) was found to be an intermediate product of DMP biodegradation and accumulated in the culture solution, which caused a sharp decrease in pH of medium and inhibited both the growth of alga and the biodegradation of DMP. The role of IC for improving the biodegradation of DMP was both to supply a favorite carbon source to support the rapid growth of alga and to mitigate the decrease of pH because of the production of PA. A suggested second-order kinetic equation of organic pollutant biodegradation by microalgae (-dC/dt = KNr) fitted well with the experimental data and the correlation coefficients were all above 0.9. The second-order constant (K) apparently declined with the increase of initial IC because lower ratio between organic carbon from DMP and IC was used to support the growth of alga when initial IC increased.

Biodegradation, Environmental↗