PubMed Health⌕ Search

Biomedical subjects

Y Song

Publications and source records attributed to Y Song.

At least 37 records · Page 2Linked to original sources
PubMed2005

Enhanced tissue strength in cryopreserved, collagen-based blood vessel constructs.

Traditional cryopreservation methods do not adequately preserve complex natural or engineered multicellular tissues due to the ice formation in the extracellular matrices. Vitrification is an alternate ice-free method for cryopreservation. This study compares the effects of vitrification and conventional cryopreservation on an engineered blood vessel construct. Collagen-based vascular constructs were used as models in this study. Tubular constructs were cut into rings and distributed into fresh, frozen, and vitrified groups for evaluation of mechanical properties and cell viability. Passive mechanical tests revealed enhanced tissue strength after both freezing and vitrification. Cryosubstitution studies of frozen and vitrified constructs revealed negligible ice in the vitrified specimens and extensive ice formation in the extracellular matrix of frozen specimens. Morphological changes associated with ice formation were visible within tissues preserved using traditional cryopreservation but not in tissue preserved using vitrification. The metabolic assay results indicated that vitrified tissue had similar viability to fresh controls. These results suggest that the increased tissue strength after cryopreservation may relate to thermal property change during preservation that cross-link collagen in tissue-engineered blood vessels. Further development of this cryopreservation method is necessary to minimize the alteration in material property and maintain cell viability of the constructs.

Animals↗
PubMed2005

The 825C/T polymorphism of G-protein beta3 subunit gene and risk of ischaemic stroke.

In this study, we investigated the association of G protein beta3 subunit gene (GNB 3) C825T polymorphism with ischaemic stroke and its subtypes in the Chinese Han population in a large case-control study. A total of 990 ischemic stroke patients and 1124 controls were recruited from six medical centres in China. Genotype was determined by polymerase chain reaction and restriction fragment length polymorphism (RFLP) assay. Logistic regression analysis was performed to identify the independent risk factors for stroke. The frequency of 825T carriers is significantly higher in cerebral thrombosis in male subjects (OR=1.35, 95% CI, 1.01-1.82, P=0.046). After further adjustment with traditional risk factors to stroke, the association is not significant. In conclusion, the GNB3 825T allele is not an independent risk factor to ischaemic stroke in the Chinese population.

Aged↗
PubMed2005

Deciphering cancer complexities in genetically engineered mice.

Because the pRb pathway is disrupted in most solid human cancers, we have generated genetically engineered mouse cancer models by inactivating pRb function in several cell types, including astrocytes and mammary, prostate, ovarian, and brain choroid plexus epithelia. In every case, proliferation and apoptosis are acutely induced, predisposing to malignancy. Cell type dictates the pathways involved in tumor progression. In the astrocytoma model, we developed strategies to induce events in the adult brain, either throughout the tissue or focally. Both K-Ras activation and Pten inactivation play significant roles in progression. In the prostate model, adenocarcinoma progression depends on Pten inactivation. However, nonautonomous induction of p53 in the mesenchyme leads to evolution of both compartments, with p53 loss occurring in the mesenchyme. Thus, studies in these models continue to identify key tumorigenesis mechanisms. Furthermore, we are hopeful that the models will provide useful preclinical systems for diagnostic and therapeutic development.

Animals↗
PubMed2005

Immobilization and condensation of DNA with 3-aminopropyltriethoxysilane studied by atomic force microscopy.

We used different methods to modify a mica surface with 3-aminopropyltriethoxysilane (APTES), and then used it as substrate to immobilize DNA for atomic force microscopy (AFM) observation. The evaporation method and solution modifying method were investigated and evaluated. The solution modifying method was found to be relatively simple and effective. Using an APTES solution-modified mica surface, DNA immobilization appeared more reproducible and it could be imaged in liquid. The mixed solution of APTES and DNA was dropped directly onto the mica surface for AFM imaging. We found that DNA can condense in APTES water solutions. Toroids, rods and intermediate structures of condensation were captured by AFM.

DNA↗
PubMed2005

Haplotyping of TNFalpha gene promoter using melting temperature analysis: detection of a novel -856(G/A) mutation.

Tumor necrosis factor alpha (TNFalpha) is a potent cytokine with a wide range of pro-inflammatory activities and plays a critical role in the pathogenesis of a number of infectious, inflammatory, autoimmune, and metabolic diseases. We determined four single-nucleotide polymorphisms (SNPs), -1031(C/T), -863(C/A), -857(C/T), and -308(G/A) in the TNFalpha promoter region using melting temperature analysis, among 1451 geriatric autopsy samples. Two adjacent SNPs, -863(C/A) and -857(C/T), were directly assayed by a single probe reaction, which correctly determined three of four expected haplotypes. Sequence confirmation related that the most rare haplotype (8/2902 chromosomes, frequency: 0.26%) contains a novel mutation of -856(G/A), instead of the predicted haplotype. These results indicate that melting temperature analysis provides a robust method to determine the polymorphisms in the TNFalpha promoter.

Haplotypes↗
PubMed2004

Control of retinoic acid synthesis and FGF expression in the nasal pit is required to pattern the craniofacial skeleton.

Endogenous retinoids are important for patterning many aspects of the embryo including the branchial arches and frontonasal region of the embryonic face. The nasal placodes express retinaldehyde dehydrogenase-3 (RALDH3) and thus retinoids from the placode are a potential patterning influence on the developing face. We have carried out experiments that have used Citral, a RALDH antagonist, to address the function of retinoid signaling from the nasal pit in a whole embryo model. When Citral-soaked beads were implanted into the nasal pit of stage 20 chicken embryos, the result was a specific loss of derivatives from the lateral nasal prominences. Providing exogenous retinoic acid residue development of the beak demonstrating that most Citral-induced defects were produced by the specific blocking of RA synthesis. The mechanism of Citral effects was a specific increase in programmed cell death on the lateral (lateral nasal prominence) but not the medial side (frontonasal mass) of the nasal pit. Gene expression studies were focused on the Bone Morphogenetic Protein (BMP) pathway, which has a well-established role in programmed cell death. Unexpectedly, blocking RA synthesis decreased rather than increased Msx1, Msx2, and Bmp4 expression. We also examined cell survival genes, the most relevant of which was Fgf8, which is expressed around the nasal pit and in the frontonasal mass. We found that Fgf8 was not initially expressed along the lateral side of the nasal pit at the start of our experiments, whereas it was expressed on the medial side. Citral prevented upregulation of Fgf8 along the lateral edge and this may have contributed to the specific increase in programmed cell death in the lateral nasal prominence. Consistent with this idea, exogenous FGF8 was able to prevent cell death, rescue most of the morphological defects and was able to prevent a decrease in retinoic acid receptorbeta (Rarbeta) expression caused by Citral. Together, our results demonstrate that endogenous retinoids act upstream of FGF8 and the balance of these two factors is critical for regulating programmed cell death and morphogenesis in the face. In addition, our data suggest a novel role for endogenous retinoids from the nasal pit in controlling the precise downregulation of FGF in the center of the frontonasal mass observed during normal vertebrate development.

Acyclic Monoterpenes↗
PubMed2004

The role of implant alignment on stability and particles on periprosthetic osteolysis--A rabbit model of implant failure.

The study objective was to determine the tissue response to polyethylene and/or titanium particles and the role that these play in peri-prosthetic osteolysis in a rabbit model of implant failure. Twenty-two mature rabbits were used. Unilateral tibial arthroplasty was performed on all of them. The test animals received implants that were intentionally rotationally unstable with reference to the host tibia in order to create a model of failure. The test rabbits were divided into three groups. Group 1 consisted of seven rabbits in which only the carrier was implanted. Group 2 consisted of seven rabbits that received only polyethylene particles suspended in the carrier. Group 3 consisted of eight rabbits that received a mixture of polyethylene and titanium alloy particles suspended in the carrier. The rabbits were sacrificed at 6 months post surgery. The entire knee, together with the immediately surrounding soft tissue, was retrieved. The position of the implant in each rabbit was assessed with reference to its alignment to the tibia. The number of inflammatory, foreign-body reactive cells, the presence of neovascularization, edema, and necrosis in the periprosthetic zones were recorded and assessed in a qualitative and semiquantitative manner. Quantitative histomorphometry was used to determine the proportion of implant surface that interfaced with osseous or fibrous tissue. Also assessed was the thickness and maturity of the fibrous tissue and the endosteal remodeling activity in the peri-implant bone counting both osteoclastic and osteoblastic activity. The results showed that implanted particles and misalignment of the implants combined to produce peri-prosthetic bone resorption. Bone resorption was found to be proportional to the degree of misalignment. The animals that received combined polyethylene/titanium particles had a greater degree of foreign-body and inflammatory response with osteolysis than the other groups. The combination of bio-material particles (polyethylene and titanium alloy) produced a greater degree of bone resorption than the single biomaterial particles (polyethylene). The amount of bone resorption surrounding the implant was directly proportional to the degree of misalignment of the implant.

Alloys↗
PubMed2004

Ionic pair complexes with well-separated columnar stack structure based on [Pt(mnt)2]- ions showing unusual magnetic transition: syntheses, crystal structures, and magnetic properties.

Three ion pair complexes, [4-R-benzylpyridinium][bis(maleodinitriledithiolato)platinum(III)] (abbreviated as [RBzPy][Pt(mnt)(2)]; R = Cl (1), Br (2), or NO(2) (3)), have been synthesized. The cations and anions stack into well-separated columns in the solid state, and the Pt(III) ions form a 1-D zigzag chain within a [Pt(mnt)(2)](-) column through Pt...S, S...S, and Pt...S...Pt interactions. The chain is uniform in 1 and 2, while it alternates in 3. Unusual magnetic phase transitions from paramagnetism to diamagnetism were observed in these three complexes at approximately 275 K for 1, approximately 269 K for 2, and approximately 184 K for 3. These phase transitions were also found in DSC measurements for 1 and 2. The overall magnetic behaviors for 1-3 indicate the presence of antiferromagnetic exchange interactions in the high-temperature phase and spin-gapped systems in the low-temperature phase. Below 50 K, 2 exhibits weak ferromagnetism. The spontaneous moments are nearly repressed by a field of 1.0 T. The crystal structure of 2 at 173 K reveals that there are two crystallographically independent [Pt(mnt)(2)](-) entries in an asymmetric unit. These two crystallographically independent [Pt(mnt)(2)](-) entries satisfy the spin-canting condition, and the EPR spectra measured at room temperature exhibit anisotropic character. Therefore, the weak ferromagnetic behavior in the low-temperature region for 2 can be attributed to the spin-canting phenomenon.

Journal Article↗
PubMed2004

Characterization of development-related genes for the cestode Bothriocephalus acheilognathi.

Differential gene expression of mature and immature Bothriocephalus acheilognathi cestodes was analyzed using the suppression subtractive hybridization technique. Five mature-associated cDNAs were isolated and characterized. Virtual Northern blot and RT-PCR analyses confirmed that four of the five genes were upregulated in mature parasites. The sequence analysis revealed that one gene encoded the structural protein chorion precursor, and the three encoded functional proteins homologous to yolk ferritin, sodium/hydrogen exchanger and muscin-like protein. Another gene appeared to be specific to B. acheilognathi, encoding a putative metal-bound protein. Although results obtained in the present study are preliminary, the information about the five genes may provide clues for further investigation on the decline in parasite numbers during the maturation of B. acheilognathi.

Amino Acid Sequence↗
PubMed2004

Homology modeling of lanosterol 14alpha-demethylase of Candida albicans and Aspergillus fumigatus and insights into the enzyme-substrate Interactions.

The crystal structure of 14alpha-sterol demethylase from Mycobacterium tuberculosis (MT_14DM) provides a good template for modeling the three dimensional structure of lanosterol 14alpha-demethylase, which is the target of azole antifungal agents. Homologous 3D models of lanosterol 14alpha-demethylase from Candida albicans (CA_14DM) and Aspergillus fumigatus (AF_14DM) were built on the basis of the crystal coordinates of MT_14DM in complex with 4-phenylimidazole and fluconazole. The reliability of the two models was assessed by Ramachandran plots, Profile-3D analysis, and by analyzing the consistency of the two models with the experimental data on the P450(14DM). The overall structures of the resulting CA_14DM model and AF_14DM model are similar to those of the template structures. The two models remain the core structure characteristic for cytochrome P450s and most of the insertions and deletions expose the molecular surface. The structurally and functionally important residues such as the heme binding residues, the residues lining the substrate access channel, and residues in active site were identified from the model. To explore the binding mode of the substrate with the two models, 24(28)-methylene-24,25-dihydrolanosterol was docked into the active site of the two models and hydrophobic interaction and hydrogen-bonding were found to play an important role in substrate recognition and orientation. These results provided a basis for experiments to probe structure-function relationships in the P450(14DM). Although CA_14DM and AF_14DM shared similar core structural character, the active site of the two models were quite different, thus allowing the rational design of specific inhibitors to the target enzyme and the discovery of novel antifungal agents with broad spectrum.

Amino Acid Sequence↗
PubMed2004

Estimating the relative risk of developing melanoma in INK4A carriers.

Estimation of the relative risk of cancer due to rare germline mutations using population-based epidemiological techniques is challenging, since studies with very large numbers of subjects are required. In this pilot study using a novel study design, we evaluated the role of INK4A mutations in melanoma by comparing patients with multiple primary melanomas to those with single primaries. Patients were ascertained from the Surgery and Dermatology Clinics at Memorial Sloan-Kettering Cancer Center and at the Yale University Pigmented Lesion Clinic. Subjects completed a questionnaire covering risk factors for melanoma and were tested for INK4A mutations. Five (8%) of 65 patients with multiple primaries had a mutation, compared with none of 88 patients with single primaries (P=0.03). Examination of other factors, such as number of nevi on the arms of the patients, fair skin, hair and eye colour, and other phenotypic characteristics associated with the risk of melanoma, demonstrates that these factors exhibit higher prevalence in the multiple primary cases than in the single primaries. These results provide evidence of the utility of the new study design in evaluating the impact of rare but highly penetrant cancer risk factors.

Adolescent↗
PubMed2003

Sphingolipid-cholesterol domains (lipid rafts) in normal human and dog thyroid follicular cells are not involved in thyrotropin receptor signaling.

Partition of signaling molecules in sphingolipid-cholesterol-enriched membrane domains, among which are the caveolae, may contribute to signal transduction efficiency. In normal thyroid, nothing is known about a putative TSH/cAMP cascade compartmentation in caveolae or other sphingolipid-cholesterol-enriched membrane domains. In this study we show for the first time that caveolae are present in the apical membrane of dog and human thyrocytes: caveolin-1 mRNA presence is demonstrated by Northern blotting in primary cultures and that of the caveolin-1 protein by immunohistochemistry performed on human thyroid tissue. The TSH receptor located in the basal membrane can therefore not be located in caveolae. We demonstrate for the first time by biochemical methods the existence of sphingolipid-cholesterol-enriched domains in human and dog thyroid follicular cells that contain caveolin, flotillin-2, and the insulin receptor. We assessed a possible sphingolipid-cholesterol-enriched domains compartmentation of the TSH receptor and the alpha- subunit of the heterotrimeric G(s) and G(q) proteins using two approaches: Western blotting on detergent-resistant membranes isolated from thyrocytes in primary cultures and the influence of 10 mm methyl-beta-cyclodextrin, a cholesterol chelator, on basal and stimulated cAMP accumulation in intact thyrocytes. The results from both types of experiments strongly suggest that the TSH/cAMP cascade in thyroid cells is not associated with sphingolipid-cholesterol-enriched membrane domains.

Animals↗
PubMed2003

Deletion of the Hsp70 chaperone gene SSB causes hypersensitivity to guanidine toxicity and curing of the [PSI+] prion by increasing guanidine uptake in yeast.

Yeast Ssb proteins (Ssbp) are ribosome-associated Hsp70 chaperones that function in translation. Elevated levels of Ssbp enhance the ability of over-expressed Hsp104 chaperone to eliminate the yeast [PSI+] prion, while depletion of Ssbp reduces this effect. Millimolar concentrations of guanidine in the growth medium cure yeast cells of prions by inactivating Hsp104. Guanidine is also toxic to yeast, irrespective of the status of Hsp104 and [PSI+]. Strains that lack Ssbp are hypersensitive to guanidine toxicity. Here we show that ssb- cells have normal numbers of [PSI+] "seeds", but can be cured of [PSI+] using one-sixth of the guanidine concentration required to eliminate [PSI+] from SSB cells. Correspondingly, the level of intracellular guanidine was eight-fold higher in ssb- cells than in wild-type cells, which explains all effects of Ssbp depletion on susceptibility to guanidine. The sensitivity of wild-type cells to the effects of guanidine also correlated with guanidine uptake, which was enhanced at low temperature. Guanidine sensitivity of strains mutated in any of 16 ABC membrane transporters, which are implicated in multidrug resistance, was normal. We found that an erg6 mutant that has an altered membrane lipid composition was hypersensitive to guanidine toxicity, but the lipid composition of ssb- cells was identical to that of wild-type cells. Our results suggest that Ssbp depletion does not affect prion seed regeneration, and that elevated guanidine uptake by ssb- cells may be due to increased retention rather than to an alteration in active or passive transport of the compound.

Cell Division↗
PubMed2003

Aquaporin-1 deletion reduces osmotic water permeability and cerebrospinal fluid production.

Aquaporin-1 (AQP1) is a water channel that is strongly expressed at the ventricular-facing surface of choroid plexus epithelium. Using wildtype and AQP1 null mice, we developed novel methods to compare the water permeability in isolated choroid plexus, and cerebrospinal fluid (CSF) production in living mice. Osmotically-induced water transport was rapid in freshly isolated choroid plexus from wildtype mice as measured by a spatial-filtering optical method, and reduced by 5-fold by AQP1 deletion. CSF production, an isosmolar fluid secretion process, was measured by a dye dilution method involving fluid collections using a second microneedle introduced into the cisterna magna. CSF production in wildtype mice was (in microl/min) 0.37 +/- 0.04 microl/min (control), 0.16 +/- 0.03 microl/min (acetazolamide-treated) and 1.14 +/- 0.15 microl/min (forskolin-treated), and reduced by up to 25% in AQP1 null mice. The impaired CSF production in AQP1 null mice provides direct functional evidence for the involvement of AQP1 in CSF formation.

Animals↗
PubMed2003

Co-delivery of IL-2 or liposomes augment the responses of mice to a DNA vaccine for pseudorabies virus IE180.

Recently we have demonstrated, with a DNA vaccine, that the immediate early protein (IE180) of pseudorabies virus provides a moderate level of protection in mice. In order to improve its immunogenicity and protective capacity, this IE180 DNA vaccine was delivered to C3H/HeJ mice either in combination with an IL-2 expressing plasmid or complexed with cationic liposomes. Co-delivery of the vaccine and IL-2 DNA by gene gun resulted in seroconversion in 5/5 of the vaccinated mice after a single administration, whereas two intramuscular (i.m.) injections were required to achieve seroconversion in all mice. Antibody and delayed-type hypersensitivity responses were augmented in mice, which received the DNA vaccine and the IL-2 gene compared to those of mice receiving the DNA vaccine alone. In addition, the time of death after challenge was significantly delayed in mice, which received the IL-2 gene. The proportion of surviving mice (40%), however, was similar to that obtained in mice which received the vaccine alone by gene gun. Liposome-mediated vaccine delivery also resulted in a higher rate of seroconversion when compared with that induced by the naked DNA vaccine. Thus, all vaccinated mice seroconverted after either two i.v. or three i.m. injections of the liposome/DNA complex, with 40 and 25% of these mice being protected against challenge, respectively. These data support that co-administration of the IE180 DNA vaccine with the IL-2 gene or delivery in liposomes are two effective approaches to increase its immunogenicity.

Animals↗
PubMed2002

Relationship between the stability of lysozymes mutated at the inside hydrophobic core and secretion in Saccharomyces cerevisiae.

The relationship between the stability of lysozymes mutated at the inside hydrophobic core and secretion was investigated to understand the optimal secretion of mutant lysozymes in the Saccharomyces cerevisiae. S91T mutant lysozyme increased in the methyl residue inside the core greatly increased the conformational stability. The secretion amount of S91T in S. cerevisiae increased greatly compared with wild-type lysozyme. On the other hand, I55V and T40S/I55V mutant lysozymes decreased in methyl residue inside the core brought about their unstable conformation. The secretion amounts of these unstable mutant lysozymes significantly decreased. In addition, the effect of glycosylation on the secretion of these mutants was investigated. The secretion amounts of glycosylated lysozyme S91T/G49N with stable hydrophobic core greatly increased compared with that of glycosylated lysozyme G49N, while those of mutant I55V/G49N and T40S/I55V/G49N with unstable hydrophobic core greatly decreased. These results indicate that the secretion amounts of mutant lysozymes increase in proportion to the hydrophobic core stabilities and that a similar good correlation was obtained with glycosylated lysozymes.

Anti-Infective Agents↗
PubMed2002

Antiinflammatory screening of the medicinal plant Gynura procumbens.

Gynura procumbens is used in Thai folk medicine to treat topical inflammation, rheumatism, and viral ailments. In the present work, attempts were made to verify the folk medicinal claim that the crude ethanolic extract of G. procumbens has antiinflammatory action and to relate the activity to particular fractions using a croton oil-induced mouse ear inflammation model. The original ethanolic extract of G. procumbens was partitioned between water and ethyl acetate. The residues were subjected to antiinflammatory evaluation. While the water extract did not show any antiinflammatory activity, the administration of the original organic extract significantly inhibited the increase in ear thickness in response to croton oil (n = 5). The activity of 0.75 mg/ear original organic extract showed similar antiinflammatory activity (inhibition 65.2%) to that of 6 mg/ear hydrocortisone 21-hemisuccinate sodium salt (inhibition 64.8%). The organic extract was then fractionated with a series of solvents in order of increasing polarity. Each fraction was dried, dissolved in acetone and monitored using the same bioassay. These experiments showed that the hexane and toluene fractions showed significant inhibitions of 44.6% and 34.8%, respectively. These two fractions had similar activities to 4 mg/ear of hydrocortisone (inhibition 35.0%). The possible chemical constituents in the extracts and fractions were investigated using thin layer chromatography and specific color reagents. These tests showed that steroids might be one class of antiinflammatory compounds in this plant.

Animals↗