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Biomedical subjects

B Li

Publications and source records attributed to B Li.

At least 55 records · Page 3Linked to original sources

Molecular characterization of HMW glutenin subunit allele 1Bx14: further insights in to the evolution of Glu-B1-1 alleles in wheat and related species.

1Bx14 is a member of the high molecular weight (HMW) glutenin subunits specified by wheat Glu-B1-1 alleles. In this work, we found that the full-length amino acid sequence of 1Bx14 and 1Bx20, the last two of the three cysteine residues, which are conserved in 1Bx7, 1Bx17 and homologous 1Ax and 1Dx subunits, were replaced by tyrosine residues. In the 5' flanking regions (-900 to -1,200 bp relative to the start codon), a novel miniature inverted-repeat transposable element insertion was present in 1Bx12 and 1Bx20 but not 1Bx7 and 1Bx17. 1Bx14 and 1Bx20 like alleles were readily found in tetraploid wheat subspecies but not several S genome containing Aegilops species. Phylogenetic analysis showed that the four molecularly characterized Glu-B1-1 alleles (1Bx7, 1Bx14, 1Bx17, 1Bx20) could be divided into two allelic lineages. The lineage represented by 1Bx7 and 1Bx17 was more ancient than the one represented by 1Bx14 and 1Bx20. Combined, our data establish that 1Bx14 and 1Bx20 represent a novel subclass of Glu-B1-1 alleles. Based on current knowledge, potential mechanism involved in the differentiation of two Glu-B1-1 lineages is discussed.

Alleles↗

Delaying acute graft-versus-host disease in mouse bone marrow transplantation by treating donor cells with antibodies directed at l-selectin and alpha4-integrin prior to infusion.

Acute graft-versus-host disease (GVHD) is still a major hurdle for successful bone marrow transplantation (BMT). Although many immunosuppressive drugs are available, none of them alone or in combination are able to completely abolish acute GVHD. The lifelong immunosuppression profoundly reduces the quality of life of BMT recipients. Therefore, new therapeutic approaches are needed. We previously reported that, in an acute GVHD model using SCID mice as recipient, incubating donor spleen cells with antibodies directed at CD49d and CD62L could significantly delay the occurrence of acute GVHD. To test the potential usefulness of this treatment in BMT, we examined this therapeutic protocol in a mouse BMT model. The present mouse BMT study confirmed our previous results that incubation of donor cells with antibodies directed at CD49d and CD62L prior to infusion into the recipient can effectively delay acute GVHD, allowing the recipients to recover from the side effects of total body irradiation. This one-time treatment is easy and simple and may be modified for clinical usage.

Animals↗

Elicitation of both CD4 and CD8 T-cell-mediated specific immune responses to HCA587 protein by autologous dendritic cells.

We recently cloned a new member of cancer/testis antigen named HCA587, which was highly expressed in human hepatocellular carcinoma (HCC) tissues. To investigate it as a potential tumour-specific target for immunotherapy, the immunogenicity of this protein, especially the ability to induce specific cellular immune responses, was evaluated in the present study. As dendritic cells (DC) are the most potent antigen-presenting cells, DC-based vaccination has recently shown marked promise for the treatment of human malignancies by immunological intervention. Here, we demonstrate that autologous DC loaded with HCA587 protein could induce specific T-cell responses in healthy individuals by in vitro stimulations. Enzyme-linked immunospot analysis for interferon-gamma (IFN-gamma) secretion demonstrated HCA587-specific CD8(+) T cells in the antigen-stimulated peripheral blood lymphocytes, and the analysis of CD4(+) T cells by proliferation assay also showed antigen-specific reactivities in normal donors. Two-colour flow cytometric analysis of surface markers and intracellular cytokine expression demonstrated that HCA587-specific cytotoxic T lymphocytes exhibited a heterogeneous CD8(+)/CD56(+) expression, and a striking T-helper 1 cytokine bias (IFN-gamma(high)/IL-4(low)) was observed for both CD4(+) and CD8(+) HCA587-specific lymphocyte populations. We conclude that HCA587 is a potent immunogen that can induce CD4(+) and CD8(+) T-cell-mediated specific immune responses, and these findings propose HCA587 as a good candidate for the development of a therapeutic protein-based DC tumour vaccine for the treatment of HCC patients.

Antigens, Neoplasm↗

Haplotypic analyses of the aldosterone synthase gene CYP11B2 associated with stage-2 hypertension in northern Han Chinese.

The objective of this study was to investigate the association of polymorphisms in the aldosterone synthase gene CYP11B2 (-344T/C, Lys173/Arg, and an intronic conversion [IC]) with stage-2 hypertension in northern Han Chinese. A total of 503 hypertensives and their age-, gender-, and area-matched controls were included in this study. The female hypertensives had significantly higher frequencies of the -344T, 173Lys, and IC-conversion alleles (p = 0.002, 0.002, and 0.014, respectively). The estimated frequency of haplotype composed of the -344T, 173Lys, and IC-conversion alleles (haplotype 4) was significantly higher in the female hypertensives compared with their controls (p = 0.007). Using a multivariate score test, we found that haplotype 4 remained associated with female hypertension after the adjustment for covariates (p = 0.003), while the haplotype 3 of T-Arg-WT showed a protective effect both in the males and in the females (p = 0.03 and 0.006, respectively). The odds ratio for haplotype phase of 4-4 was 2.60 (95% CI, 1.21-5.58) and for 3-3, 0.20 (95% CI, 0.03-0.77). These results indicate that the Lys173 and the IC-conversion allele of the CYP11B2 gene confer an increased risk for stage-2 hypertension in northern Han Chinese women.

Asian People↗

Creating chemical diversity to target protein kinases.

Protein kinases play crucial roles in regulating virtually every cellular process and are currently attracting tremendous interest as drug targets from the pharmaceutical industry. The major challenges facing the development of the potential kinase inhibitor drugs are: selectivity, physical properties (solubility, molecular weight), and pharmacological properties (bioavailability, half life, toxicity, etc.) This review focuses on how selective protein kinase inhibitors that target the ATP and allosteric binding sites are currently being identified and optimized.

Animals↗

Studies on a poly(styrene-divinylbenzene)-encapsulated sub-micrometer sized organic-inorganic hybrid silica packing, its synthesis, some of its characteristics and applications.

The hydrolyzed product of a mixture of tetraethoxysilane (TEOS) and vinyltriethoxysilane (VTEOS) was used to produce organic-inorganic hybrid silica by sol-gel processing. The product was a kind of composite with vinyl groups incorporated into the structure of the final particles through C-Si bonds. These hybrid silica particles were spherical and the average diameter was about 350 nm as measured by scanning electron microscopy (SEM). The particles were non-porous as proved by BET method. The hybrid silica particles were subsequently encapsulated with a layer of PS-DVB, a polymeric mixture of styrene and divinylbenzene, with the vinyl groups on the particle surface as anchors of the polymer chains. The encapsulated packing was used as stationary phases in CEC, and the chromatographic behaviors were studied. Experimental results showed that, as expected, the smaller diameter stationary phase resulted in higher column efficiencies. In comparison with the columns packed with 5 microm diameter encapsulated-silica stationary phases of the same kind and other sub-micron stationary phases reported, it can be concluded that the EOF velocity is independent of the particle diameter when using 350 nm phase, but determined by the surface nature of the stationary phase.

Microscopy, Electron, Scanning↗

Genetic mapping in (Populus tomentosa x Populus bolleana) and P. tomentosa Carr. using AFLP markers.

The AFLP genetic linkage maps for two poplar cultivars were constructed with the pseudo-test-cross mapping strategy. The hybrids were derived from an interspecific backcross between the female hybrid clone "TB01" (Populus tomentosa x Populus bolleana) and the male clone "LM50" ( P. tomentosa). A total of 782 polymorphic fragments were obtained with a PCR-based strategy using 49 enzyme-nested ( EcoRI/ MseI) primer combinations. Six hundred and thirty two of these fragments segregated in a 1:1 ratio ( P<0.01), indicating that these DNA polymorphisms are heterozygous in one parent and null in the other. The linkage analysis was performed using Mapmaker version 3.0 with LOD 5.0 and a maximum recombination fraction (theta) of 0.3. Map distances were estimated using the Kosambi mapping function. In the framework map for "LM50" (P. tomentosa), 218 markers were aligned in 19 major linkage groups. The linked loci spanned approximately 2,683 cM of the poplar genome, with an average distance of 12.3 cM between adjacent markers. For "TB01" (P. tomentosa x P. bolleana), the analysis revealed 144 loci, which were mapped to 19 major linkage groups and covered about 1,956 cM, with an average distance of 13.6 cM between adjacent markers. These maps covered about 87% and 77% of the estimated genome size of parents "LM50" and "TB01", respectively. The maps developed in this study lay an important foundation for future genomics research in poplar, providing a means for localizing genes controlling economically important traits in P. tomentosa.

Chromosome Mapping↗

Shoot production per responsive leaf explant increases exponentially with explant organogenic potential in Nicotiana species.

As part of the effort to develop optimal plant varieties for the production and molecular farming of plant-made pharmaceuticals, this study evaluated shoot organogenic potential of a total of 115 Nicotiana accessions, representing 53 species. To induce shoots, leaves from seedling grown in vitro were cut into pieces, cultured on shoot-induction medium under low light for 3 weeks, and then subcultured onto the same medium for another 4 weeks under normal light. Statistical analysis detected significant differences among the 115 accessions for the percentage of leaf explants producing shoots and the number of shoots produced per responsive leaf explant. Importantly, regression analysis also found an exponential relationship between the number of shoots produced per responsive leaf explant and the percentage of leaf explants producing shoots. The number of shoots produced per responsive leaf explant increased rather slowly, ranging from zero to around five, as the percentage of leaf explants producing shoots increased from 0 to 80%, but the increase became dramatic as the percentage increased from 80% to 100%, reaching as high as 35 shoots per responsive leaf explant. This exponential relationship is the first of its kind to be established in plant regeneration studies using either organogenesis or somatic embryogenesis systems. A possible mechanism that governs the establishment of the exponential relationship is discussed.

Culture Techniques↗

Expression of butyrate response factor 1 in HTLV-1-transformed cells and its transactivation by tax protein.

Tax oncoprotein of Human T-lymphotropic virus 1 (HTLV-1) has been proposed to dysregulate the expression of a number of cellular genes, many of which play a critical role for cell proliferation. Our initial data demonstrated that the immediate-early gene butyrate response factor 1 ( BRF1) was upregulated in HTLV-1-infected cells. The ensuing studies revealed that the effect of Tax was mediated through two transcription elements. The more proximal element, located in the vicinity of TATA box, accounted for the main Tax transactivating effect, and it appeared to be a novel transcription factor-binding site. It involved the CCTCCTC sequence (nt -59/-53, relative to transcription start site) and was dubbed BRF1 Tax-responsive site (BTRS). The cellular protein(s) recruited into the formation of DNA-protein complex at this binding site were not identified. The other element, located further upstream, was a consensus cAMP-responsive site (CRE) TGACGTCA, spanning positions -400 to -393. CRE-binding protein (CREB) was found to mediate the transactivating effect of Tax at this site. Our results present the first evidence that the Tax transactivator has a capability to modulate the expression of BRF1 and that this effect is mediated by CRE and a novel BTRS motifs.

Cell Line↗

Optimization of RNA yield, purity and mRNA copy number by treatment of urine cell pellets with RNAlater.

BACKGROUND: We have shown that measurement of mRNA for cytotoxic attack proteins perforin and granzyme B in urinary cells is a noninvasive means of diagnosing acute rejection of human renal allografts. Urinary cell mRNA studies have yielded useful information in other patient populations such as patients with cancer. The isolation of sufficient and high quality ribonucleic acid (RNA) from urinary cells however is problematic. RNAlater, an RNA stabilization solution, has been reported to optimize RNA isolation from tumor tissues stored at room temperature and from pigment-rich ocular tissues. METHODS: We explored whether the addition of RNAlater to urine cell pellets improves RNA yield, enhances purity and facilitates measurement of low abundance mRNAs. We measured, with the use of real-time quantitative polymerase chain reaction (PCR) assay, levels of expression of a constitutively expressed gene 18S rRNA and mRNA for granzyme B and transforming growth factor-beta(1) (TGF-beta(1)) in urine specimens and renal biopsies obtained from renal allograft recipients. RESULTS: RNA yield (P<0.01, Wilcoxon signed rank test) and the A260/A280 ratio (P<0.01) were both higher with urine cell pellets treated with RNAlater prior to snap freezing compared to cell pellets that were not treated with RNAlater prior to snap freezing. Levels (copy number per 1 microg of total RNA) of 18S rRNA (P<0.02), granzyme B mRNA (P=0.002) and TGF-beta(1) (P=0.02) were all higher with treated urine cell pellets compared to untreated cell pellets. Kruskall-Wallis one way analysis of variance and pair-wise comparisons with Student-Newman-Keuls test showed that the levels of mRNA for granzyme B (P<0.05) and TGF-beta(1) (P<0.05) are significantly different between renal allograft biopsies and untreated urine cell pellets but not between the biopsy specimens and RNAlater-treated urine cell pellets. CONCLUSIONS: The addition of RNAlater to urine cell pellets improves RNA isolation from urinary cells and facilitates measurement of low abundance mRNAs.

Adolescent↗

Genotype-phenotype analysis for the polymorphic CA repeat in the insulin-like growth factor-I (IGF-I) gene.

Polymorphic microsatellite dinucleotide (cytosine-adenine, CA) repeats of the insulin-like growth factor-I (IGF-I) gene may have implications in the development of certain types of cancer and osteoporosis. We studied correlations between IGF-I genotypes determined by direct sequencing and plasma IGF-I levels among 113 healthy individuals (60 men and 53 women), who were originally enrolled as controls for hospital-based case-control studies of breast and prostate cancer. On the contrary to an earlier observation, there were no differences in plasma IGF-I levels between those with and without the allele of 19 repeats. With adjustment for other confounders, there were no trends in plasma IGF-I levels with increasing or decreasing the number of CA repeats among all study subjects combined, all Whites or all Blacks, suggesting no overall functional significance of this polymorphism. Opposite trends observed in gender and racial subgroups, i.e., an inverse association between plasma IGF-I levels and the CA repeat length in white women and a positive association in black men, are likely to be chance findings.

Alleles↗

Diploid-dependent regulation of gene expression: a genetic cause of abnormal development in fish haploid embryos.

A diploid-dependent regulatory mechanism of gene expression for spatial patterning of the eye in vertebrates has been determined by analyzing the phenotypes of haploid goldfish embryos. There are two gene loci in charge of eye spatial patterning during embryonic morphogenesis. The expressional probability for each copy of the two genes in a set of chromosomes is 50%. A pair of genes in two sets of homologous or heterologous chromosomes is 100% and essential for normal gene expression. The haploid condition itself would result in the obstruction of gene expression and abnormal development because the diploid-dependent regulatory apparatus will regulate gene expression in a haploid embryo according to the same rule as in the diploid embryo.

Animals↗

The effects of the farnesyl transferase inhibitor FTI L-778,123 on normal, myelodysplastic, and myeloid leukemia bone marrow progenitor proliferation in vitro.

The effects of the farnesyl transferase inhibitor FTI-778,123 on the proliferation of normal, MDS, AML, and CML hemopoietic progenitor cells was studied. MDS myeloid and erythroid progenitors are significantly more sensitive to FTI than normal progenitors while AML myeloid progenitors may be somewhat more sensitive than normal progenitors. In contrast, no difference between CML and normal progenitors are detectable. These data strongly suggest that a trial of this agent in patients with MDS and perhaps in patients with AML is indicated.

Alkyl and Aryl Transferases↗

T-bet expression is upregulated in active Behçet's disease.

AIM: To investigate T-bet mRNA and protein expression on peripheral blood mononuclear cells (PBMC) in patients with Behçet's disease with active uveitis. METHODS: Blood samples were taken from 24 patients with Behçet's disease who had active uveitis and 16 healthy individuals. PBMC were subjected to analysis of T-bet mRNA and protein expression using semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR) and western blot, respectively. The products from PCR were sequenced. In order to determine the influence of activation on T-bet expression, the phytohaemagglutinin (PHA) stimulated PBMC from each sample were also evaluated for expression of T-bet mRNA and protein. RESULTS: A significantly increased T-bet mRNA accumulation was detected in the samples from patients with active Behçet's disease compared with that in controls. A 62 kDa band was detectable in patients with active Behçet's disease, but not in controls. No difference was found between patients with Behçet's disease who had active uveitis and normal controls concerning the expression of either T-bet mRNA or its protein after stimulation with PHA for 72 hours. CONCLUSION: Behçet's disease is associated with an upregulation of T-bet expression, which supports a role for the Th1 subset of T cells in the pathogenesis of this disease.

Adult↗

Structure-function dynamics and modeling analysis of the micro-environment of activated sludge floc.

Biodegradation by microorganisms and mass transfer resistance in the micro-environment of activated sludge floc can cause changes in substrate and dissolved oxygen concentrations within the floc and can contribute to stratification of microbial processes inside the flocs. In this study, an integrated model of the microenvironment of the activated sludge floc was developed for floc from wastewaters from several sources and of varying strengths for dynamic simulation of the combined biological processes of COD and nitrogen removal. The model simulation results and measured profiles show the heterogeneous and gradient-governed microenvironment of activated sludge floc under different substrate and bulk oxygen concentrations. The substrate concentration increase zones inside the floc were present in all activated sludge floc from the Miller Brewing Co. wastewater treatment facility (high pollutant strength), with an oxygen penetration depth of only 0.15 mm into the outer layer. The anoxic and substrate concentration increase zones also dominated in the activated sludge floc from the Mill Creek Plant influent (medium pollutant strength), with the outer layer (0.20 mm) participating in the metabolism of the pollutants. The radius of the substrate concentration increase zone inside the sludge floc decreased with pollutant removal along the length of the tank. When the pollutant concentration in the bulk wastewater was low (Muddy Creek Plant), the substrate concentration increase zone disappeared; the whole floc was aerobic and in a high redox status. Our experiments and model analyses demonstrate that the microorganisms' structure-functions inside activated sludge floc change with the bulk substrate concentration and dissolved oxygen concentration.

Biodegradation, Environmental↗

RNA-dependent RNA polymerase activity of Classical swine fever virus NS5B protein expressed in natural host cells.

The NS5B gene, cloned from Classical swine fever virus (CSFV) genome, was expressed in porcine kidney cells PK-15, natural host of CSFV. In purifying cytoplasmic extracts from these cells by means of different concentrations of salt, glycerol and detergent four fractions, namely crude supernatant (SC) and different purified supernatants (S1, S2 and S3) were obtained. Using Western blot analysis the NS5B protein was found in all these fractions, showing that it was soluble in both higher and lower concentrations of salt, glycerol and detergent. The NS5B protein present in the four different fractions exhibited RNA-dependent RNA polymerase (RdRp) activity, but it was unable to complete the whole process of RNA synthesis. Site-directed mutation analysis showed that Thy216 and Cyt228 were essential for RNA synthesis while Cyt219 was not, suggesting that CSFV RdRp was template-specific. We conclude that initiation of RNA synthesis by CSFV RdRp includes also template priming.

Animals↗

Functional impact of lymphangiogenesis on fluid transport after lymph node excision.

When a lymph node is excised, lymphangiogenesis occurs to maintain flow in the affected area. However, a complex network of small vessels replaces the node and these newly formed vessels might increase resistance to lymph transport. To test this in sheep, the popliteal lymph node from one hind limb was removed surgically. The contralateral node was left intact. After 4 to 6 weeks (a period that allowed regenerated vessels to restore flow), a prenodal lymphatic vessel in each limb was cannulated with a polyethylene catheter to permit saline infusion into the node or lymphatic regeneration site. Infusion pressures were monitored from t-pieces inserted between the infusion pump and the point of entry of the catheters in the prenodal ducts. We observed that the flow rate versus perfusion pressure relationships were significantly different in the 2 experimental preparations (node intact limbs, n = 13; node excised limbs, n = 10). In the limbs undergoing lymphangiogenesis, much higher infusion pressures were required to generate a given flow rate. Additionally, the regenerated lymphatic network provided a significantly increased resistance to flow. The data suggested that lymphangiogenesis restored fluid continuity to some extent in the area occupied originally by the popliteal lymph node. However, the transport properties exhibited by the newly formed lymphatics were insufficient to restore flow parameters to their original state.

Animals↗

Copper speciation and accumulation in the gill microenvironment of carp ( Cyprinus carpio) in the presence of kaolin particles.

Carp were exposed to copper adsorbed on kaolin particles at various concentrations (0-2.4 mg/L), and net accumulation due to elevated adsorbed copper in the surrounding water was observed. Copper speciation in the water and fish gill microenvironment was modeled in the presence and absence of kaolin using a chemical speciation program (MINTEQA2). The adsorption affinity constants of kaolin for copper at various pH values used in the speciation calculations were experimentally determined, and the quantitative relationship between the affinity constant and pH was modeled. Copper accumulations in fish gills exposed to kaolin-adsorbed copper was then observed. The results indicate that desorption occurred in the fish gill microenvironment due to both mucus competition of copper and slight increase in water pH. Furthermore, the available copper species increased as a result of desorption, causing a net accumulation of Cu by the gills.

Adsorption↗