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Biomedical subjects

L Sun

Publications and source records attributed to L Sun.

547 records · Page 31Linked to original sources

Analogs of guanine nucleoside triphosphates for sequencing applications.

We have synthesized more than 30 different deoxyribonucleosides and triphosphates with modifications either in the base or the phosphate moiety as analogs of 2'-dGTP for DNA sequencing applications. All the modified nucleoside triphosphates were tested as substrates for DNA polymerases, including Sequenase T7 DNA polymerase or Thermo Sequenase DNA polymerase. Two of the analogs, 7-ethyl-7-deaza-dGTP and 7-hydroxymethyl-7-deaza-dGTP meet our requirements as better sequencing reagents.

Guanine Nucleotides↗

Magnetic flow sorting using a model system of human lymphocytes and a colloidal magnetic label.

Cells of identical physical properties that differ in the expression of surface proteins can be sorted conveniently using immunospecific stains conjugated to fluorescent, or magnetic, labels. Immunomagnetic cell sorting using commercial batch sorters offers advantages of high sorting capacity, high viability of sorted fractions, and high depletion rates; its disadvantages are low enrichment rate and batch processing. The authors developed and tested a continuous, flow-through magnetic cell sorter for small volume, experimental cell enrichment. Freshly isolated human peripheral lymphocytes were labeled using an immunofluoromagnetic sandwich consisting of mouse anti human CD8 monoclonal antibody-fluorescein conjugate and rat anti mouse polyclonal antibody-colloidal iron-dextran conjugate. A total of 2-3 min lymphocytes were sorted per hour using a saturation magnetic field of 1.334 T and a five channel sorter. The fluorescent cells were distributed among the channels in relation to their fluorescence intensity and magnetic susceptibility. The purity (68-85%) and enrichment rates (16-34x) were comparable to those of commercial batch magnetic separators; sorting capacity and recovery of the enriched fractions (up to 32%) were limited by the small scale of the sorter. Future direction is focused on increasing the resolution, recovery, and sorting capacity of the enriched fractions, and testing the sorter on other cell systems.

Animals↗

Rapid cell isolation by magnetic flow sorting for applications in tissue engineering.

Rapid and efficient cell sorting methods are important for tissue progenitor cell isolation. We built and evaluated a laboratory prototype of a continuous flow, quadrupole magnetic cell sorter. The sorter was tested on a model cell system of human peripheral lymphocytes. The helper T cell subpopulation was targeted by primary, mouse anti-CD4 monoclonal antibody conjugated to a fluorochrome (FITC), and magnetized by secondary, anti-FITC antibody magnetic colloid. The purities and recoveries of the cell fractions were measured by flow cytometry and an automated cell counter. Cells were spread across the flow according to their magnetophoretic mobilities. The purity of the CD4 cell enriched fraction was 99.6%, and the purity of the CD4 cell depleted fraction was 2% for an initial CD4 cell purity of 36%; the corresponding recovery of the enriched CD4 cell fraction was 59% at a sorting speed of 4,200 cells/s (four experiments). The recovery could be increased to 90% with a concomitant decrease in the purity of CD4 cell enriched fraction to 66%. This type of sorting should be applicable to any cells in suspension for which a suitable antibody exists, in particular, to large, fragile cells.

Antibodies, Monoclonal↗

Phased array RF coils for high-resolution MRI of the inner ear and brain stem.

OBJECTIVE: The spatial resolution in MRI is predominantly limited by the available signal-to-noise ratio (S/N). To increase the S/N, a four coil phased array consisting of bilateral pairs of semioval coils was constructed for high resolution imaging of the temporal bone and brainstem. MATERIALS AND METHODS: Coil sizes of 10 x 6 and 6 x 4.5 cm were tested. The S/N values were measured in vivo and with homogeneous phantoms and compared to commercial 3 in receive and quadrature head coils. RESULTS: At a depth of 4-5 cm, phantom studies yielded S/N improvements of a factor of 1.26-1.37 with the large array compared to the 3 in coil and 2.33-1.74 compared to the head coil. Similar improvements (1.16 and 2.37) were obtained in inner ear images. No further improvement was achieved at this depth with the small array. At a depth of 8 cm, phantom studies yielded similar S/N for the quadrature head coil and two bilaterally placed large array coils, while a factor of 1.27 was obtained in brainstem images. CONCLUSION: The use of phased array coils yields significantly increased S/N and is thus valuable for high resolution MRI.

Brain Stem↗

Prevention of induced prostate-related cancer by soy protein isolate/isoflavone-supplemented diet in Lobund-Wistar rats.

Based on epidemiological surveys, the low incidence of clinical prostate cancer among aged men in Japan and China were attributed to high consumption of soybean-derived food in which phytoestrogens have numerous anticancer mechanisms. The prostate model in Lobund-Wistar (L-W) rats produce high levels of testosterone (T). They are inherently predisposed to develop induced and spontaneous metastasizing adenocarcinomas, which are (T)-dependent in early stages and T-independent in advanced stages. In the experiment reported here, 2 groups of L-W rats (age 2 months) were fed soy-containing diets: (a) commercial diet L-485 (TekLad) with soy meal; or (b) a soy-free diet (L-474) in which casein was replaced by soy protein isolate/isoflavones (SPII). At age 3 months, all rats were inoculated i.v. with MNU; and 14 months later, 17/58 (29.3%) of rats on diet L-485 developed cancer in avg 12 months, compared to 5/50 (10%) of rats on the SPII diet in avg 12.1 months (P = 0.001). In the latter rats, the serum levels of T, and weights of testes were significantly reduced; but in the former rats, serum levels of T remained elevated, suggesting that soy meal in L-4,85 blocked the estrogenic effects of phytoestrogens.

Adenocarcinoma↗

[Identification of a novel human MAST4 gene, a new member of the microtubule associated serine-threonine kinase family].

Human protein kinases make up a large superfamily of homologous proteins, which are related by virtue of their kinase domains (also known as catalytic domains). Here we report the cloning and characterization of a novel human MAST4 (microtubule associated serine/threonine kinase family member 4) gene, which locates on human chromosome 5q13. The MAST4 cDNA is 7587 base pairs in length and encodes a putative protein of 2435 amino acids which contains a serine/threonine kinase domain and a PDZ domain. MAST4 protein has 64%, 63%, 59% and 39% identical aminoacid residues with MAST1, MAST2, MAST3 and MASTL respectively. RT-PCR analysis revealed relatively high expression level of MAST4 in most normal human tissues, with an exception of in testis, small intestine, colon and peripheral blood leukocyte.

Amino Acid Sequence↗