Biomedical subjects

S Li

Publications and source records attributed to S Li.

At least 415 records · Page 23Linked to original sources

PubMed2000

Molecular cytogenetic analysis of eight inversion duplications of human chromosome 13q that each contain a neocentromere.

Neocentromeres are fully functional centromeres that have arisen in previously noncentromeric chromosomal locations on rearranged chromosomes. The formation of neocentromeres results in the mitotic stability of chromosomal fragments that do not contain endogenous centromeres and that would normally be lost. Here we describe a unique collection of eight independent patient-derived cell lines, each of which contains a neocentromere on a supernumerary inversion duplication of a portion of human chromosome 13q. Findings in these patients reveal insight into the clinical manifestations associated with polysomy for portions of chromosome 13q. The results of FISH and immunofluorescent analysis of the neocentromeres in these chromosomes confirm the lack of alpha-satellite DNA and the presence of CENtromere proteins (CENP)-C, -E, and hMAD2. The positions of the inversion breakpoints in these chromosomes have been placed onto the physical map of chromosome 13, by means of FISH mapping with cosmid probes. These cell lines define, within chromosome 13q, at least three distinct locations where neocentromeres have formed, with five independent neocentromeres in band 13q32, two in band 13q21, and one in band 13q31. The results of examination of the set of 40 neocentromere-containing chromosomes that have thus far been described, including the 8 neocentromere-containing chromosomes from chromosome 13q that are described in the present study, suggest that chromosome 13q has an increased propensity for neocentromere formation, relative to some other human chromosomes. These neocentromeres will provide the means for testing hypotheses about sequence requirements for human centromere formation.

Aneuploidy↗

PubMed2000

The immunosuppressive macrolide RAD inhibits growth of human Epstein-Barr virus-transformed B lymphocytes in vitro and in vivo: A potential approach to prevention and treatment of posttransplant lymphoproliferative disorders.

Whereas the standard immunosuppressive agents foster development of posttransplant lymphoproliferative disorders (PTLDs), the impact of RAD, a macrolide with potent immunosuppressive properties, and other immunosuppressive macrolides on these disorders remains undetermined. We found that RAD had a profound inhibitory effect on in vitro growth of six different PTLD-like Epstein-Barr virus+ lymphoblastoid B cell lines. Similar to normal T cells, RAD blocked cell-cycle progression in PTLD-like B cells in the early (G(0)/G(1)) phase. Furthermore, RAD increased the apoptotic rate in such cells. The drug also had a profound inhibitory effect on the growth of PTLD-like Epstein-Barr virus+ B cells xenotransplanted s.c. into SCID mice. The degree of the RAD effect varied among the three B cell lines tested and was proportional to its effects on the cell lines in vitro. In this in vivo xenotransplant model, RAD markedly delayed growth or induced regression of the established tumors. In one line, it was able to eradicate the tumor in four of eight mice. When RAD treatment was initiated before tumor cell injection, a marked inhibition of tumor growth was seen in all three lines. In two of them, the drug prevented tumor establishment in approximately 50% of mice (5/11 and 5/8). In summary, RAD is a potent inhibitor of PTLD-like cells in vitro and in vivo. These findings indicate that, in contrast to the standard immunosuppressive agents, macrolides such as RAD may be effective in prevention and treatment of PTLDs.

Animals↗

PubMed2000

Effects of soaking temperature and soaking time during preparation of water extract of tea on anticlastogenicity against environmental tobacco smoke in the sister-chromatid exchange assay.

Water extract of tea (WET) was prepared by soaking green tea at different temperatures for various periods of time and was used to test whether the soaking temperature and soaking time during the preparation of WET influence the content of polyphenols and the anticlastogenicity of WET against environmental tobacco smoke (ETS). Five major polyphenols in WET were measured. Extractable-respirable particulate (ERP) was obtained from ETS-contaminated indoor air (ERP-ETS). The sister-chromatid exchange assay (SCE) was utilized to evaluate the clastogenic effects of ERP-ETS and the anticlastogenic effects of WET. The results indicate that ERP-ETS is clastogenic and WET has significant anticlastogenic effects on ERP-ETS. The content of polyphenols and the anticlastogenic potential of WET depended on the soaking temperature and soaking time during WET preparation. At the soaking temperature of 80 degrees C, an increased soaking time was correlated with a higher percentage of polyphenols and a concomitantly enhanced anticlastogenic efficacy. By contrast, at the soaking temperature of 100 degrees C, a longer soaking time was associated with a higher percentage of polyphenols concomitant with a lower anticlastogenic efficacy. The data suggest that, besides polyphenols, and additional material(s), which may be partially inactivated at 100 degrees C, is contributing to the anticlastogenicity of WET.

Antimutagenic Agents↗

PubMed2000

Effect of environmental tobacco smoke on intracellular free calcium of boar sperm incubated in seminal plasma.

The extractable-respirable particulate of environmental tobacco smoke (ERP-ETS) was collected and tested for its effect on the intracellular free Ca(2+) concentration, [Ca(2+)](i), in boar sperm incubated in seminal plasma in vitro. The intracellular free Ca(2+) level is reportedly closely related to in sperm capacitation and acrosome reaction, which must take place to confer upon sperm the full fertilizing potential. In the absence of ERP-ETS [Ca(2+)](i) in the boar sperm increased dramatically from initially 75 nM to about 248 nM after a 5-h incubation in seminal plasma. However, such an increase was virtually inhibited by ERP-ETS at a concentration of 10 microg/ml, without any significant reduction of the sperm motility and viability. We conclude that application of ERP-ETS negatively affected the [Ca(2+)](i) of boar sperm in vitro.

Acrosome Reaction↗

PubMed2000

[Effects of electromagnetic radiation from handsets of cellular telephone on neurobehavioral function].

In order to study the effects of electromagnetic radiation from handsets of cellular telephone on neurobehavioral function, 81 staff with handsets of cellular telephone and 63 staff without handsets of cellular telephone from corporations were selected as the subjects. The subjects were investigated by questionnaire on their general health, lifestyle habit, suppress of spirit, handset using of cellular telephone, environmental exposure, morbidity, and the neurobehavioral core test battery(NCTB). The data was analyzed by chi-square, stepwise regression analysis and covariance statistics. The results showed that the average reaction time in user's group was longer than that in control group (P < 0.01). The time of using handset was negatively associated with corrected reaction number (P < 0.01). The fast reaction time and the slowest reaction time were positively associated with the length of handset using (P < 0.01, P < 0.05). The results suggested that the handset using could cause adverse health effects in neurobehavioral function.

Behavior↗

PubMed2000

Cytokine-induced Src homology 2 protein (CIS) promotes T cell receptor-mediated proliferation and prolongs survival of activated T cells.

Members of the suppressor of cytokine signaling (SOCS) family were discovered as negative regulators of cytokine signaling by inhibition of the Janus kinase-signal transducer and activator of transcription (Jak-STAT) pathway. Among them, cytokine-induced Src homology 2 (SH2) protein (CIS) was found to inhibit the interleukin 3- and erythropietin-mediated STAT5 signaling pathway. However, involvement of SOCS proteins in other signaling pathways is still unknown. This study shows that the expression of CIS is selectively induced in T cells after T cell receptor (TCR) stimulation. In transgenic mice, with selective expression of CIS in CD4 T cells, elevated CIS strongly promotes TCR-mediated proliferation and cytokine production in vitro, and superantigen-induced T cell activation in vivo. Forced expression of CIS also prolongs survival of CD4 T cells after TCR activation. Molecular events immediately downstream from the TCR are not changed in CIS-expressing CD4 T cells, but activation of mitogen-activated protein (MAP) kinase pathways by TCR stimulation is significantly enhanced. Together with the increased MAP kinase activation, a direct interaction of CIS and protein kinase Ctheta was also demonstrated. These results suggest that CIS is one of the important regulators of TCR-mediated T cell activation. The functions of CIS, enhancing TCR signaling and inhibiting cytokine signaling, may be important in the regulation of immune response and homeostasis.

Animals↗

PubMed2000

Structural basis of hyaluronan degradation by Streptococcus pneumoniae hyaluronate lyase.

Streptococcus pneumoniae hyaluronate lyase (spnHL) is a pathogenic bacterial spreading factor and cleaves hyaluronan, an important constituent of the extra- cellular matrix of connective tissues, through an enzymatic beta-elimination process, different from the hyaluronan degradation by hydrolases in animals. The mechanism of hyaluronan binding and degradation was proposed based on the 1.56 A resolution crystal structure, substrate modeling and mutagenesis studies on spnHL. Five mutants, R243V, N349A, H399A, Y408F and N580G, were constructed and their activities confirmed our mechanism hypothesis. The important roles of Tyr408, Asn349 and His399 in enzyme catalysis were proposed, explained and confirmed by mutant studies. The remaining weak enzymatic activity of the H399A mutant, the role of the free carboxylate group on the glucuronate residue, the enzymatic behavior on chondroitin and chondroitin sulfate, and the small activity increase in the N580G mutant were explained based on this mechanism. A possible function of the C-terminal beta-sheet domain is to modulate enzyme activity through binding to calcium ions.

Amino Acid Sequence↗

PubMed2000

Involvement of ERK in BMP-2 induced osteoblastic differentiation of mesenchymal progenitor cell line C3H10T1/2.

The signaling mechanisms responsible for bone morphogenetic protein (BMP) induced osteoblast differentiation remains poorly understood. Previous research demonstrated that Smad proteins are the substrates and the mediators of BMP bound serine/threonine receptor kinase. In the present study, we examined the possible involvement of extracellular signal-regulated kinase (Erk) in the BMP induced osteoblast differentiation of mesenchymal progenitor cell C3H10T1/2. Our results indicate that BMP-2 inducement increased MAP kinase activity in mesenchymal progenitor cell line C3H10T1/2. Contrary to previous reports, this increased MAP kinase activity showed a latent but sustained pattern. Elevation of Erk1 and Erk2 protein levels was observed simultaneously. RT-PCR results demonstrated that the elevation of Erk protein level in BMP-2 induced cells was from the upregulation of mRNA expression. Furthermore, upregulated Erk proteins present enhanced phosphorylation. By using a dominant-negative Erk2 cell line, we demonstrated that nonfunctional Erk2 partially eliminated BMP-2 induced cell proliferation and ALP activity in the C3H10T1/2 cell. These results indicate that Erk is involved in BMP-2 induced osteoblast differentiation. The results also demonstrate that a latent and sustained signaling pattern exists in BMP induced signaling cascade.

Animals↗

PubMed2000

Comparison of environmental tobacco smoke concentrations and mutagenicity for several indoor environments.

Environmental tobacco smoke (ETS) is a major source for indoor air pollution. Although ETS-caused indoor air pollution has been well studied in the developed countries, few studies have examined ETS indoor air pollution in China, which currently has the largest population of tobacco smokers. In this study, respirable-particulate (RP) from ETS-contaminated (RP-ETS) indoor air was collected and measured in 5 different indoor environments during the winter in the northwestern Liaoning Province, China. The extractable portion of RP-ETS (ERP-ETS) was obtained by dichloromethane extraction and used in the Salmonella mutagenicity assay in the presence of S9 using strains TA98, TA100, and TA1538. The percentage of RP-ETS attributable to ETS (ETS-RP) and the percentage of ERP-ETS attributable to ETS (ETS-ERP) were estimated by measuring the concentration of solanesol, an ETS marker. Comparative results in 5 different indoor environments were: (1) the concentration of RP-ETS ranged from 197.3 to 1227.6 microg/m(3) and approximately 64.7 to 92. 0% of the RP-ETS originated from ETS; (2) the concentration of ERP-ETS ranged 88.8 to 601.5 microg/m(3) and approximately 83.1 to 95.4% of the ERP-ETS originated from ETS; (3) the mutagenic potency (revertants/m(3)) of ERP-ETS ranged from 60.4 to 595.5 for TA98, from 33.7 to 312.8 for TA100, and from 49.7 to 475.2 for TA1538. The data indicate that the extent of ETS pollution and the potential health hazards of ETS to humans in the five indoor environments are in the following increasing order: rural bedrooms, urban living rooms, office rooms, restaurants, and passenger cars in that area.

Air Pollution, Indoor↗

PubMed2000

Modulation of transcription factor AP-1 activity in murine EL-4 thymoma cells by vomitoxin (deoxynivalenol).

Trichothecene mycotoxins have been reported to suppress or superinduce cytokine mRNA expression by leukocytes both in vitro and in vivo. Modulation of transcription factor activities may be critical for these observations. Here, the effect of trichothecene vomitoxin (VT, deoxynivalenol) on activator protein-1 (AP-1) activity was determined in the murine EL-4 thymoma. Electrophoretic mobility shift assay (EMSA) revealed that VT modulated AP-1 binding activity in a concentration- and time-dependent manner when using a synchronous model in which VT was added concurrently with phorbol 12-myristate 13-acetate (PMA) and ionomycin (ION) to EL-4 cells. Induction of AP-1 binding activity by PMA/ION was suppressed in the presence of VT for a short period (1 to 12 h), but was enhanced upon prolonged VT exposure (48 to 72 h). VT also enhanced AP-1 binding activity when added to the cell culture 12 h after PMA/ION activation (delayed synchronous model). Using specific antibodies against AP-1 complex proteins, it was demonstrated by gel supershift assay that VT preferentially affected phosphorylated c-Jun, Jun B, c-Fos, and Fra-2 binding activities, whereas it did not alter Jun D and Fra-1 binding. A transient transfection assay demonstrated that these increased binding activities are associated with enhanced AP-1 transactivation potential. Elevation of AP-1 activity may contribute to cytokine dysregulation and immunotoxic effects associated with exposure to trichothecene mycotoxins such as VT.

Animals↗

PubMed2000

Down-regulation of the endoplasmic reticulum chaperone GRP78/BiP by vomitoxin (Deoxynivalenol).

The mechanisms by which trichothecene mycotoxins cause immunological effects in leukocytes such as cytokine up-regulation, aberrant IgA production, or apoptotic cell death are not fully understood. In the present study, mRNA differential display analysis was used to evaluate changes in gene expression induced by the trichothecene vomitoxin (VT or deoxynivalenol) in a T-cell model, the murine EL-4 thymoma, that was stimulated with phorbol 12-myristate 13-acetate (PMA) and ionomycin (ION). Ten differentially expressed fragments of cDNA were isolated and sequenced and three of these were identified as the known genes GRP78/BiP, P58(IPK), and RAD17. Most notably, expression of GRP78/BiP (a 78-kDa glucose-regulated protein), a stress-response gene induced by agents or conditions that adversely affect endoplasmic reticulum (ER) function, was found to decrease in VT-exposed cells. Competitive RT-PCR analysis revealed that 250 ng/ml VT decreased GRP78/BiP mRNA expression in both unstimulated and PMA/ION-stimulated EL-4 cells at 6 and 24 h after VT treatment. Western blotting confirmed that VT (50 to 1000 ng/ml) also significantly diminished GRP/BiP protein levels in a dose-response manner in PMA/ION-stimulated cells. GRP78/BiP has been shown to play a role in regulation of protein folding and secretion, and to protect cells from apoptosis. When PMA/ION-stimulated cells were incubated with 50 to 1000 ng/ml VT for 24 h, 200-bp DNA laddering, a hallmark of apoptosis, increased in a dose-dependent manner. In addition to GRP78, mRNA expression of the cochaperone P58(IPK), which is the 58-kDa cellular inhibitor of the double-stranded RNA-regulated protein kinase (PKR), was also shown to be suppressed by VT-treatment. GRP78 and P58(IPK) are critical for maintenance of cell homeostasis and prevention of apoptosis. The down-regulation of these molecular chaperones by VT represent a novel observation and has the potential to impact immune function at multiple levels.

Animals↗

PubMed2000

Improvement of the enantioselectivity of lipase-catalyzed naproxen ester hydrolysis in organic solvent.

A method is presented to improve the enantioselectivity of lipase-catalyzed hydrolysis of naproxen methyl ester in water-saturated isooctane. It is shown that coupling of the enantioselective hydrolysis of Naproxen methyl ester with the photo-dissociation methanol leads to the photocatalytic conversion of methanol into water, by which the equilibrium constant (K) of the lipase-catalyzed hydrolysis was changed. The equilibrium yield and enantiomeric excess are increased. Because the lipase would not dissolve in the organic solvent, it was adsorbed on photocatalyst particles, which may facilitate the isolation of enzyme from reaction system.

Journal Article↗

PubMed2000

Mechanisms of ionotropic glutamate receptor-mediated excitotoxicity in isolated spinal cord white matter.

Spinal cord injury involves a component of glutamate-mediated white matter damage, but the cellular targets, receptors, and ions involved are poorly understood. Mechanisms of excitotoxicity were examined in an in vitro model of isolated spinal dorsal columns. Compound action potentials (CAPs) were irreversibly reduced to 43% of control after 3 hr of 1 mM glutamate exposure at 37 degrees C. AMPA (100 microM) and kainate (500 microM) had similar effects. Antagonists (1 mM kynurenic acid, 10 microM NBQX, 30 microM GYKI52466) were each equally protective against a glutamate challenge, improving mean CAP amplitude to approximately 80% versus approximately 40% without antagonist. Joro spider toxin (0.75 microM), a selective blocker of Ca(2+)-permeable AMPA receptors, was also protective to a similar degree. Ca(2+)-free perfusate virtually abolished glutamate-induced injury ( approximately 90% vs approximately 40%). MK-801 (10 microM) had no effect. Glutamate caused damage (assayed immunohistochemically by spectrin breakdown products) to astrocytes and oligodendrocytes consistent with the presence of GluR2/3 and GluR4 in these cells. Myelin was also damaged by glutamate likely mediated by GluR4 receptors detected in this region; however, axon cylinders were unaffected by glutamate, showing no increase in the level of spectrin breakdown. These data may guide the development of more effective treatment for acute spinal cord injury by addressing the additional excitotoxic component of spinal white matter damage.

Animals↗

PubMed2000

Congenital absence of permanent teeth in a six-generation Chinese kindred.

We report on rare, heritable, permanent tooth agenesis in a large Chinese kindred. The congenital absence of permanent teeth except the first and second accessory teeth was observed in 52 individuals through six successive generations in the kindred comprising 328 members. Clinical assessments were carried out, and inheritance mode and spousal influence of the anomaly on their offspring were analyzed. Consequently, the anomaly was transmitted in an autosomal dominant fashion with incomplete penetrance (P = 0.88), and no significant clinical manifestations other than the oligodontia were found. A geographical or environmental effect on the affected individuals was obviously eliminated, because any who are related to the kindred but live under the same conditions are fully healthy. The disorder we describe, therefore, differs from any previously reported oligodontia/anodontia syndromes. The oligodontia ranged from a few teeth to the whole set of teeth, and usually occurred at a period from age 7 or 8 years, the time when primary teeth are normally replaced by permanent teeth, to the forties. Roentgenography of the affected persons indicated that only the first and/or second accessory teeth with tooth buds developed as permanent teeth. In fact, the diphyodontic germination sometimes occurred in the oral cavity of the affected individuals.

China↗

PubMed2000

[Effects of glutamate and glutamine on learning and memory of rats].

The effects of glutamate (Glu) and glutamine (Gln) on learning and memory of rats and the mechanism involved were examined. Three groups of rats were fed with stock diet (control group), and the diets supplemented with 2% Glu (Glu group) or 2% Gln (Gln group) respectively. Compared with control group, the active avoidance response rats of Gln group in shuttle box test was significantly elevated while the time of passive avoidance response shortened. The contents of brain free amino acids, such as Gln, Glu, aspartic acid and arginine, activity of brain nitric oxide synthase (NOS), Bmax of N-methyl-D-aspartate(NMDA) receptors in hippocampus of Gln group were all increased significantly. Such changes were not observed in Glu group. The results suggested that Gln could improve the learning ability and memory of rats, which might be related to the change of amino acids composition of brain, the synthesis of messenger nitric oxide and the binding capacity of NMDA receptor in hippocampus.

Animals↗

PubMed2000

Tapasin is required for efficient peptide binding to transporter associated with antigen processing.

The transporter associated with antigen processing (TAP) binds peptides in its cytosolic part and subsequently translocates the peptides into the lumen of the endoplasmic reticulum (ER), where assembly of major histocompatibility complex (MHC) class I and peptide takes place. Tapasin is a subunit of the TAP complex and binds both to TAP1 and MHC class I. In the absence of tapasin, the assembly of MHC class I in the ER is impaired, and the surface expression is reduced. To clarify the function of tapasin in the processing of antigenic peptides, we studied the interaction of peptide and TAP, peptide transport across the membrane of the ER, and association of peptides with MHC class I molecules in the microsomes derived from tapasin mutant cell line 721.220, its sister cell line 721.221 expressing tapasin, and their HLA-A2 transfectants. The binding of peptides to TAP in tapasin mutant 721.220 cells was significantly diminished in comparison with 721.221 cells. Impaired peptide-TAP interaction resulted in a defective peptide transport in tapasin mutant 721.220 cells. Interestingly, despite the diminished peptide binding to TAP, the transport rate of TAP-associated peptides was not significantly altered in 721.220 cells. After transfection of tapasin cDNA into 721.220 cells, efficient peptide-TAP interaction was restored. Thus, we conclude that tapasin is required for efficient peptide-TAP interaction.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

PubMed2000

High-level rearrangement and transcription of yeast artificial chromosome-based mouse Ig kappa transgenes containing distal regions of the contig.

The mouse Ig kappa L chain gene locus has been extensively studied, but to date high-level expression of germline transgenes has not been achieved. Reasoning that each end of the locus may contain regulatory elements because these regions are not deleted upon V kappa-J kappa joining, we used yeast artificial chromosome-based techniques to fuse distal regions of the contig to create transgene miniloci. The largest minilocus (290 kb) possessed all members of the upstream V kappa 2 gene family including their entire 5' and 3' flanking sequences, along with one member of a downstream V kappa 21 gene family. In addition, again using yeast artificial chromosome-based technology, we created Ig kappa miniloci that contained differing lengths of sequences 5' of the most distal V kappa 2 gene family member. In transgenic mice, Ig kappa miniloci exhibited position-independent and copy number-dependent germline transcription. Ig kappa miniloci were rearranged in tissue and developmental stage-specific manners. The levels of rearrangement and transcription of the distal and proximal V kappa gene families were similar to their endogenous counterparts and appeared to be responsive to allelic exclusion, but were differentially sensitive to numerous position effects. The minilocus that contained the longest 5' region exhibited significantly greater recombination of the upstream V kappa 2 genes but not the downstream V kappa 21 gene, providing evidence for a local recombination stimulating element. These results provide evidence that our miniloci contain nearly all regulatory elements required for bona fide Ig kappa gene expression, making them useful substrates for functional analyses of cis-acting sequences in the future.

Alleles↗

PubMed2000

Lyophilization of cationic lipid-protamine-DNA (LPD) complexes.

Cationic lipid-based gene delivery systems have shown promise in transfecting cells both in vitro and in vivo. However, these systems tend to form aggregates in liquid formulation during storage, which has limited their clinical applications. As a result, lyophilization of these systems has recently become a subject of increasing interest. In this paper, lyophilization of LPD, a novel cationic lipid-based gene delivery system, was studied. Both particle size and transfection efficiency could be preserved in the presence of sufficient amount of appropriate lyoprotectant. A series of monosaccharides and disaccharides, including dextrose, galactose, mannose, lactose, maltose, sucrose and trehalose, were evaluated for their lyoprotective effect and disaccharides showed more superior protection to monosaccharides. The effect of different freezing protocols for lyophilization was also evaluated and no significant difference was found. However, for freeze-thawing, fast freezing caused less aggregation. Finally, nonlyophilized LPD and LPD lyophilized with 10% sucrose were stored at different temperatures and their stability was followed for eight weeks. Lyophilized LPD could be stored at room temperature without significant change in particle size or loss of transfection efficiency.

Animals↗