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B Huang

Publications and source records attributed to B Huang.

At least 73 records · Page 4Linked to original sources
PubMed2000

Early down-regulation of K+ channel genes and currents in the postinfarction heart.

INTRODUCTION: Down-regulation of key K+ channel subunit gene expression and K+ currents is a universal response to cardiac hypertrophy, whatever the cause, including the postmyocardial infarction (post-MI) remodeled heart. METHODS AND RESULTS: We investigated the hypothesis that down-regulation of K+ channel genes and currents post-MI occurs early and before significant remodeled hypertrophy of the noninfarcted myocardium could be detected. We investigated (1) the incidence of induced ventricular tachyarrhythmias (VT) in 3-day post-MI rat heart; (2) action potential (AP) characteristics of isolated left ventricular (LV) myocytes from sham-operated and 3-day post-MI heart; (3) time course of changes in outward K+ currents Ito-fast(f) and I(K) in isolated myocytes from 3-day and 4-week post-MI noninfarcted LV and compared the changes with sham-operated animals; and (4) changes in the messenger and protein levels of Kv2.1, Kv4.2, and Kv4.3 in the LV and right ventricle of 3-day post-MI heart. Sustained VT was induced in 6 of 10 3-day post-MI rats and in none of 8 sham rats. The membrane capacitance of myocytes isolated from 3-day post-MI noninfarcted LV was not significantly different from control, whereas membrane capacitance 4-week post-MI was significantly higher, reflecting the development of hypertrophy. AP duration was increased and the density of Ito-f and I(K) were significantly decreased in 3-day post-MI LV myocytes compared with sham. The reduced density of Ito did not significantly differ in 4-week post-MI LV myocytes, whereas the density of I(K) was decreased further at 4 weeks post-MI. The changes in Ito-f and I(K) correlated with decreased messenger and protein levels of Kv4.2/Kv4.3 and Kv2.1, respectively. CONCLUSION: These results support the hypothesis that down-regulation of K+ channel gene expression and current in the post-MI LV occurs early and may be dissociated from the slower time course of post-MI remodeled hypertrophy. These changes may contribute to early arrhythmogenesis of the post-MI heart.

Action Potentials↗
PubMed2000

A minimal tiling path cosmid library for functional analysis of the Pseudomonas aeruginosa PAO1 genome.

Pseudomonas aeruginosa is an important pathogenic and environmental bacterium, with the most widely studied strain being PAO1. Using the PAO1 reference cosmid library and the recently completed PAO1 genome sequence, we have mapped a minimal tiling path across the genome using a two-step strategy. First, we sequenced both ends of a set of over 500 random and previously mapped clones to create a backbone. Second, we end-sequenced a second set of cosmid clones that were identified to lie within the larger gaps using hybridization of the reference library filters with probes designed against sequences at the center of each gap. The minimal tiling path was calculated using the program Domino (http://www.bit.uq.edu.au/download/), with the overlap between adjacent clones set to 5 kb (where possible) to minimize the chance of truncating genes. This yielded a minimal tiling cosmid library (334 clones) covering 93.7% of the genome in 57 contigs. This library has reduced to a workable set the number of clones required to represent the majority of the P. aeruginosa genome and gives the precise location of each cosmid, enabling most genes of interest to be located on clones without further screening. This library should prove a useful resource to accelerate functional analysis of the P. aeruginosa genome.

Cloning, Molecular↗
PubMed2000

Agrobacterium-mediated transformation of creeping bentgrass using GFP as a reporter gene.

Creeping bentgrass (Agrostis palustris Huds.) is a cool season grass widely used on putting greens in golf courses. Transformation of creeping bentgrass has been conducted using microprojectile bombardment and protoplast electroporation. The objective of our study is to develop an alternative and more efficient approach in transforming the grass using Agrobacterium (strain EHA 101). This technique was effective in transforming 40-day old calli derived from mature seeds cultured on MS medium supplemented with 2,4-D, kinetin, and sucrose. Dozens of transgenic plants have been produced from two independent transformed calli. Presence of functional green fluorescence protein (GFP) was detected in leaves, stems, and roots of transgenic seedlings. Four putative transgenic plants and two control plants were randomly chosen and analyzed by Southern blot analysis. Bands corresponding to the GFP gene were clearly shown in transgenic plants. These results indicated that Agrobacterium transformation can successfully be applied to creeping bentgrass.

Adenine↗
PubMed2000

Characterization of a predominant immunogenic outer membrane protein of Riemerella anatipestifer.

The ompA gene, encoding the 42-kDa major antigenic outer membrane protein OmpA of Riemerella anatipestifer, the etiololgical agent of septicemia anserum exsudativa, was cloned and expressed in Escherichia coli. Recombinant OmpA displayed a molecular mass similar to that predicted from the nucleotide sequence of the ompA gene but lower than that observed in total cell lysates of R. anatipestifer. The ompA gene showed a conserved C-terminal region comprising the OmpA-like domain and a variable N-terminal region. This structure is similar to those of the analogous outer membrane proteins of several gram-negative bacteria. However, OmpA of R. anatipestifer contains six EF-hand calcium-binding domains and two PEST regions, which distinguish it from other outer membrane proteins. The occurrence of these motifs in OmpA suggests a possible role in virulence for this protein. The ompA gene is present in the R. anatipestifer type strain and in all serotype reference strains. However, it exhibits some minor genetic heterogeneity among different serotypes, which seems not to affect the strong antigenic characteristics of the protein. OmpA is a conserved and strong antigenic determinant of R. anatipestifer and hence is suggested to be a valuable protein for the serodetection of R. anatipestifer infections, independent of their serotype.

Amino Acid Sequence↗
PubMed2000

Biodiversity of Asian rice gall midge (Orseolia oryzae Wood Mason) from five countries examined by AFLP analysis.

Amplified fragment length polymorphism (AFLP) analysis was used to assess the biodiversity of one of the most important dipteran pests of cereals, the Asian rice gall midge (Orseolia oryzae Wood Mason). Larvae and pupae were collected at 15 locations in five Asian countries and preserved in 95% ethanol for storage, shipment, and DNA extraction using cetyltrimethylammonium bromide (CTAB). Although only approximately 1 microg of DNA was extracted from a single pupa or larva, the use of several AFLP primers in various combinations meant that this amount of DNA was sufficient to allow many DNA fingerprints to be made per individual. Fingerprints were sufficiently reproducible, especially during selective amplification, to allow the genetic diversity within a field population to be characterized. Extraction of DNA from a pool of 20 insects yielded AFLP fingerprints in which variation among individuals was sacrificed in favor of detecting differences among populations. For each location, pooled DNA was amplified with three primer pairs. A total of 261 distinct AFLP bands were identified for the 45 fingerprints. Cluster analysis, performed by the unweighted pair-group method (UPGMA), separated the populations into two distinct groups. Group I included two populations from Guangdong province of southern China and one each from Laos and Imphal in northeastern India, while group II was comprised of eleven populations from elsewhere in India (Assam, Orissa, Madhya Pradesh, Andhra Pradesh, and Kerala) and from Nepal and Sri Lanka. AFLP analysis provided insight into the origins of gall midge biotypes. In 1992, the prevailing biotype in Imphal changed from Indian biotype 3 to a new biotype 3M. Our data show that biotype 3M belongs to group I and did not arise by a recent mutation from biotype 3, which belongs to group II. By contrast, Indian biotypes 2 and 4 are likely to have diverged through recent mutation and selection, as are Chinese biotypes 1 and 4. The almost simultaneous emergence of new biotypes in Kerala and Sri Lanka during 1985-1988 was most probably coincidental, because these biotypes are not closely related. AFLP fingerprints were also able to detect sexual dimorphism in the DNA of adult gall midges and to distinguish gall midge from its major parasite Platygaster oryzae.

Animals↗
PubMed2000

[ABO genotyping by PCR-direct sequencing method].

OBJECTIVE: To analyze the sequence difference between human A, B, and O alleles and establish the method of ABO genotyping by PCR direct sequencing. METHODS: PCR-direct sequencing technique was used to analyze two regions of cDNA from A transferase gene, 233-433 and 660-788. RESULTS: Two nucleotide substitutions at 258th and 297th were found in 233-433 region, and a nucleotide substitution at 700th was found in 660-788 region. At 258th, the nucleotide was guanine in A and B alleles, and adenine in O allele. At 297th, the nucleotide was adenine in A allele, and guanine in B allele. As this position, O allele was subdivided into two types, O(A) and O(G). At 700th, the nucleotide was guanine in A and O alleles, and adenine in B allele. Therefore, 8 genotypes, AA, AO(A), AB, BB, BO(G), O(A) O(A), O(G) O(G) and O(A) O(G), could be clearly determined by only analyzing the 233-433 region. The other two genotypes, AO(G) and BO(A), could be further distinguished by analyzing the 660-788 region. CONCLUSION: The technique of PCR-direct sequencing provides an effective and new method for ABO genotyping further.

ABO Blood-Group System↗
PubMed2000

[Differentiation of embryonic stem cells into neuronal cells in vitro].

OBJECTIVE: To investigate the probabilities of embryonic stem (ES) cells differentiating into neuronal cells in vitro. METHODS: Mouse ES-D3 cells were cultured on the ES conditioned media without mLIF (mouse leukemic inhibitory factor, mLIF) for 4 days until ES cell aggregations was formed (embryonic bodies, EBs). The EBs were dissociated with trypsin and exposed to retinoic acid (RA) for another 4 days, and then replated onto adhesive substrate. Immunohistochemistry and RT-PCR assay were performed to identify the properties of the differentiated cells. RESULTS: The treated cells expressed neuronal proteins such as MAP-2 and GFAP. RT-PCR assay further identified that the induced cells expressed transcripts for neural associated genes including GABA-gamma subunit receptor, brain factor-1 and tyrosine hydroxylase (TH). CONCLUSION: ES cells can be successfully induced into neuronal cells expressing multiple properties of neurons in vitro, which could provide resourceful materials for neural transplantation.

Animals↗
PubMed2000

[Influence of fluctuating ecological factors on male occurrence of Moina macrocopa].

Studies with clonal culture show that female M. macrocopa did not produce male progeny directly, or, in other words, the parthenogenetic(amictic) female could not transform into microtic(amphoteric) female, when suffered from one-off acute or durative progressive stimulation. Under durative progressive stimulation by illumination, temperature, food density or pH, the intrinsic rates of increase were 0.7748, 0.7481, 0.6539 and 0.6310 d-1 ind-1, 0.9%, 4.0%, 16.4% and 19.3% less than the control, respectively, indicating that the animal was more sensitive to the fluctuation of food density and pH followed by temperature.

Animals↗
PubMed2000

[Population density of Eucalyptus urophylla plantation].

This paper dealt with the relationships and correlation models of the population density of 5.6 years old Eucalyptus urophylla plantation with its crown width, diamter at breast height(DBH), tree height, individual standing volume, stand volume, wood properties and survital rate. The results showed that the population density remarkably affected DBH, individual standing volume, crown width, live branch height, stand volume and wood fiber width; but not affect tree height, basic density of wood, and length of wood fibers. It had a positive relationship with stand volume, live branch height and wood fibers width, and a negative relationship with DBH, individual standing volume and crown width. In addition, E. urophylla had a wide range of reasonable density. For short-rotation puplwoods, the optimum planting density of E. urophylla is 2000 individuals per hectare.

Ecology↗
PubMed2000

[Effect of modified ammonium bicarbonate on nitrification-denitrification process and NO and N2O emission].

Compared with ammonium bicarbonate(AB), the effect of modified ammonium bicarbonate (MAB) on nitrification and denitrification processes and NO and N2O emissions in a clay soil (C soil) and a loam soil (L soil) was studied in laboratory (25 degrees C and 50% WFPS). The inhibition effect of DCD from MAB on nitrification was relatively small in C soil, but considerably great in L soil. Compared with AB, MAB extended 7 days and 33 days for retaining NH4+. During 15 days, the NO emission from C soil and L soil respectively accounted for 0.60% and 1.06% of applied N under AB application (100 micrograms N.g-1), which were as 30 and 12 times as the N2O emission from corresponding soils. After applying MAB, the emission of NO from C soil and L soil decreased by 67% and 95%, and the emission of N2O decreased by 64% and 95%, respectively. After 39 days of aerobic incubation, then anaerobically flooded incubation with nitrate addition (200 micrograms KNO3-N.g-1) for 7 days, the total loss of denitrification in MAB in L soil was 50% less, and N2O emission was 113% more than in AB in same soil.

Benzidines↗
PubMed2000

[Soil moisture dynamics in forest land and wasteland of Loess Plateau].

The dynamics of soil moisture stored in a wasteland and in a forest land in the sub-humid climate region of the Loess Plateau was measured with neutron moisture meter for three successive years. Within the depth of 0-320 cm, the increment of soil water in the test 3 years was respectively -165.7 mm, -15.2 mm and -115.2 mm at the upper and lower position on north slopes and the lower position on south slope of the wasteland. Soil water increased by 17.2 mm at the upper position on north slope of the Chinese pine forest land, after > 380 mm water was consumed yearly by tree transpiration. It is suggested that forest land is essentially different from wasteland in soil water income/expense. The necessity of conducting the approaches for gaining available water of forest land was put forward.

Seasons↗
PubMed2000

[Study on the biology feature and pathogenicities to silkworm of a microsporidium isolated from Barathra brassicae L].

A microsporidium (called as Bab-M) was isolated from Barathra brassicae L. captured from suburban vegetable plot of Guang Zhou. The spores were long-ovoid in shape and 4.02 +/- 0.36 microns x 1.99 +/- 0.36 microns in size. Immunologically the microsporidium shared spore surface specific antigen(s) with N. bombycis. The ultrastructure and life cycle of Bab-M were similar to that of N. b.. The rate of transovarian transmission was high. The initial conclusion was that Bab-M should be referred to as Nosema bombycis, but there was variation between them.

Animals↗
PubMed2000

[The gaseous phase enrichment techniques in hydride generation (review)].

The gaseous phase of hydride enrichment techniques have been systematically described in this paper. These enrichment techniques including liquid nitrogen trap, balloon collection, in situ pre-concentration in pre-heated graphite furnace, trapped in absorbing solutions, and in some absorbing solid substances. The principle and the application of these techniques have been discussed detail too. The near future trends of the techniques have reviewed also.

Arsenic↗
PubMed2000

[Spectral interferences of rare earth elements with a high resolution sequential ICP-AES. IV. Spectral interferences of europium and gadolinium].

The spectral interferences of Eu and Gd(100 micrograms.mL-1, separately) as matrices on the other REEs(1 or 5 micrograms.mL-1) were studied with a high resolution sequential ICP-AES with a grating of 3,600 grooves/mm. Totally 66 prominent lines of 14 REEs were selected as the analytical lines for the spectral interference studies. The overlapping profiles of analytical lines and interference lines were obtained by scanning four solutions(blank, analyte, Eu matrix, and Gd matrix, individually) within the selected wavelength windows (0.2 nm for each analytical line). Some new emission lines of Eu and Gd were observed. The obtained information of spectral interferences would be very useful for the selection of the best analytical lines for the determination of REEs in the other REEs matrices.

Europium↗
PubMed1999

Autosomal XX sex reversal caused by duplication of SOX9.

SOX9 is one of the genes that play critical roles in male sexual differentiation. Mutations of SOX9 leading to haploinsufficiency can cause campomelic dysplasia and XY sex reversal. We report here evidence supporting that SOX9 duplication can cause XX sex reversal. A newborn infant was referred for genetic evaluation because of abnormal male external genitalia. The infant had severe penile/scrotal hypospadias. Gonads were palpable. Cytogenetic analysis demonstrated a de novo mosaic 46,XX,dup(17)(q23.1q24.3)/46, XX karyotype. Fluorescent in situ hybridization (FISH) with a BAC clone containing the SOX9 gene demonstrated that the SOX9 gene is duplicated on the rearranged chromosome 17. The presence of SRY was ruled out by FISH with a probe containing the SRY gene and polymerase chain reaction with SRY-specific primers. Microsatellite analysis with 13 markers on 17q23-24 determined that the duplication is maternal in origin and defined the boundary of the duplication to be approximately 12 centimorgans (cM) proximal and 4 cM distal to the SOX9 gene. Thus, SOX9 duplication is the most likely cause for the sex reversal in this case because it plays an important role in male sex determination and differentiation. This study suggests that extra dose of SOX9 is sufficient to initiate testis differentiation in the absence of SRY. Other SRY-negative XX sex-reversed individuals deserve thorough investigation of SOX9 gene.

Chromosomes, Human, Pair 17↗
PubMed1999

Diminished basal phosphorylation level of phospholamban in the postinfarction remodeled rat ventricle: role of beta-adrenergic pathway, G(i) protein, phosphodiesterase, and phosphatases.

Three weeks after myocardial infarction (MI) in the rat, remodeled hypertrophy of noninfarcted myocardium is at its maximum and the heart is in a compensated stage with no evidence of heart failure. Our hemodynamic measurements at this stage showed a slight but insignificant decrease of +dP/dt but a significantly higher left ventricular end-diastolic pressure. To investigate the basis of the diastolic dysfunction, we explored possible defects in the beta-adrenergic receptor-G(s/i) protein-adenylyl cyclase-cAMP-protein kinase A-phosphatase pathway, as well as molecular or functional alterations of sarcoplasmic reticulum Ca(2+)-ATPase and phospholamban (PLB). We found no significant difference in both mRNA and protein levels of sarcoplasmic reticulum Ca(2+)-ATPase and PLB in post-MI left ventricle compared with control. However, the basal levels of both the protein kinase A-phosphorylated site (Ser16) of PLB (p16-PLB) and the calcium/calmodulin-dependent protein kinase-phosphorylated site (Thr17) of PLB (p17-PLB) were decreased by 76% and 51% in post-MI myocytes (P<0.05), respectively. No change was found in the beta-adrenoceptor density, G(salpha) protein level, or adenylyl cyclase activity. Inhibition of phosphodiesterase and G(i) protein by Ro-20-1724 and pertussis toxin, respectively, did not correct the decreased p16-PLB or p17-PLB levels. Stimulation of beta-adrenoceptor or adenylyl cyclase increased both p16-PLB and p17-PLB in post-MI myocytes to the same levels as in sham myocytes, suggesting that decreased p16-PLB and p17-PLB in post-MI myocytes is not due to a decrease in the generation of p16-PLB or p17-PLB. We found that type 1 phosphatase activity was increased by 32% (P<0.05) with no change in phosphatase 2A activity. Okadaic acid, a protein phosphatase inhibitor, significantly increased p16-PLB and p17-PLB levels in post-MI myocytes and partially corrected the prolonged relaxation of the [Ca(2+)](i) transient. In summary, prolonged relaxation of post-MI remodeled myocardium could be explained, in part, by altered basal levels of p16-PLB and p17-PLB caused by increased protein phosphatase 1 activity.

Animals↗
PubMed1999

Drosophila proteins related to vertebrate DNA (5-cytosine) methyltransferases.

DNA methylation at CpG residues is closely associated with a number of biological processes during vertebrate development. Unlike the vertebrates, however, several invertebrate species, including the Drosophila, do not have apparent DNA methylation in their genomes. Nor have there been reports on a DNA (5-cytosine) methyltransferase (CpG MTase) found in these invertebrates. We now present evidence for two CpG MTase-like proteins expressed in Drosophila cells. One of these, DmMTR1, is a protein containing peptide epitopes immunologically related to the conserved motifs I and IV in the catalytic domain of the mammalian dnmt1. DmMTR1 has an apparent molecular mass of 220 kDa and, similar to mammalian dnmt1, it also interacts in vivo with the proliferating cell nuclear antigen. During interphase of the syncytial Drosophila embryos, the DmMTR1 molecules are located outside the nuclei, as is dnmt1 in the mouse blastocyst. However, DmMTR1 appears to be rapidly transported into, and then out of the nuclei again, as the embryos undergo mitotic waves. Immunofluorescent data indicate that DmMTR1 molecules "paint" the whole set of condensed Drosophila chromosomes throughout the mitotic phase, suggesting they may play an essential function in the cell-cycle regulated condensation of the Drosophila chromosomes. Through search in the genomic database, we also have identified a Drosophila polypeptide, DmMT2, that exhibits high sequence homology to the mammalian dnmt2 and the yeast CpG MTase homolog pmt1. The expression of DmMT2 appears to be developmentally regulated. We discuss the evolutionary and functional implications of the discovery of these two Drosophila proteins related to mammalian CpG MTases.

Amino Acid Sequence↗