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Biomedical subjects

F Huang

Publications and source records attributed to F Huang.

At least 163 records · Page 9Linked to original sources
PubMed1994

Thyroid hormone regulation of Na,K-ATPase alpha 2 gene expression in cardiac myocytes.

Thyroid hormone (T3) stimulates Na,K-ATPase activity and alpha and beta subunit mRNA abundances in myocardial cells in vivo and in vitro. In this study, we used transient transfection and nuclear run-on assays to determine whether T3 regulates the transcription rate of the Na,K-ATPase alpha 2 subunit gene. Primary cultures of neonatal rat cardiac myocytes were incubated with 100 nM T3 for 1, 3, and 6 d, and alpha 2 mRNA levels were measured by Northern blot hybridization analysis. There was no change in the abundance of alpha 2 mRNA by 1 d of T3 treatment, whereas a two- and threefold increase in alpha 2 mRNA was evident when cells were exposed to T3 for 3 and 6 d, respectively. A portion of the rat alpha 2 gene containing 1700 base pairs (bp) of 5'-flanking DNA sequence was isolated and fused to the firefly luciferase gene. Transient transfection experiments utilizing this chimeric gene showed no T3 trans-activation of reporter gene activity either in the absence or presence of cotransfected beta 1 or alpha 1 isoforms of rat T3 receptor (T3R). In contrast, cotransfection of T3R facilitated a strong stimulation of luciferase activity driven by a construct containing a single copy of a palindromic T3 response element (TRE). Nuclear run-on analysis indicated that the rate of transcription of the endogenous alpha 2 gene was enhanced 1.2-fold at 3 d of T3 treatment, and was not regulated at either 1 or 6 d. These results indicate that the T3-dependent increase in alpha 2 mRNA content at 6 d is mediated at a post-transcriptional level. Unexpectedly, we observed a T3-dependent three-to sixfold repression of alpha 2/luciferase expression in cardiac myocytes cotransfected with T3R. Deletion analysis of the 5' end of the alpha 2 gene revealed a negative TRE between nucleotides -354 and -100.

Animals↗
PubMed1994

The role of nm23 in transforming growth factor beta 1-mediated adherence and growth arrest.

nm23 has properties of a metastasis suppressor gene and also has been implicated in the control of response to transforming growth factor beta 1 (TGF beta 1) by studies in melanoma cells. In this report, we have examined the role of nm23 in two HT29 colon carcinoma sublines at different stages in tumor progression with different responses to TGF beta 1: the HD3 subline, which shows TGF beta 1-induced growth arrest and differentiation; and the more invasive and tumorigenic U9 subline, which induces tumors 7-fold as large as those induced by HD3 cells with one-half the latency. Analysis by semiquantitative reverse transcription-polymerase chain reaction showed that antisense phosphorothiolated oligonucleotides to the nm23 initiation site (nm23 AS oligos) decreased nm23 mRNA levels 2-8-fold in HD3 and U9 cells when normalized to beta-actin mRNA levels. However, a role for nm23 in TGF beta 1-mediated responses could only be found in HD3 cells. nm23 AS oligos inhibited the differentiation property of cell adherence over 90% in HD3 cells, and this loss of adherence could be partially blocked by concurrent treatment with TGF beta 1. In contrast, U9 cell adherence was not detectably altered by nm23 AS oligos, whether added in the presence or absence of TGF beta 1. The TGF beta 1-induced inhibition of HD3 cell proliferation was blocked by nm23 AS oligos, whereas the TGF beta 1-induced proliferation of U9 cells was unaffected by nm23 AS oligos.(ABSTRACT TRUNCATED AT 250 WORDS)

Base Sequence↗
PubMed1994

NURSETALK: the latest addition to the information highway.

NURSETALK is an electronic bulletin board system (BBS) developed to provide information and meet the communication needs of three different nursing organizations in North Carolina. It was designed and conceptualized from a user perspective to promote functionality and acceptance.

North Carolina↗
PubMed1994

The capacity for growth stimulation by TGF beta 1 seen only in advanced colon cancers cannot be ascribed to mutations in APC, DCC, p53 or ras.

Human colon cancer development is associated with the accumulation of mutations and deletions in the suppressor genes DCC, APC and p53 and mutations in the dominant oncogene K-ras, with loss of wild type alleles. In earlier studies we had observed that about half of the resected human colon cancers placed into primary culture were growth stimulated by TGF beta 1. This group included the more advanced cancers which were either poorly differentiated primary-site cancers or metastases. In contract, the more differentiated colon cancers were inhibited or unaffected by TGF beta 1, indicating that a switch in response to TGF beta 1 occurs during colon cancer progression. Different sublines of the HT29 colon carcinoma cell line model the resected cancers, responding to TGF beta 1 by proliferation, inhibition or no growth modulation. The current study shows that while the poorly differentiated, TGF beta 1-stimulated sublines are most tumorigenic, all the sublines have the same spectrum of mutations: truncating mutations in both APC (adenomatous polyposis coli) alleles, no activated ras genes, mutated and thus overexpressed p53, and very low expression of DCC compared to normal colon cells. Genes other than the four already implicated in colon carcinoma evolution are responsible for the mitogenic response to TGF beta 1 found in the more advanced cancers.

Base Sequence↗
PubMed1994

Colon carcinoma cells switch their response to transforming growth factor beta 1 with tumor progression.

Transforming growth factor beta 1 (TGF-beta 1) switches from an inhibitor of tumor cell growth to a stimulator of growth and invasion during human colon carcinoma progression. We originally observed that metastatic colon carcinoma cells in primary culture responded to TGF-beta 1 by proliferation, whereas moderate to well-differentiated primary site colon carcinomas were growth inhibited by TGF-beta 1 (P. Schroy et al., Cancer Res., 50: 261-265, 1990). We then cloned several colon carcinoma cell lines which modeled these responses to TGF-beta 1 and expressed TGF-beta 1 (M. M. Hafez et al., Cell Growth & Differ., 1: 617-626, 1990; 3: 753-762, 1992). Two of these colon carcinoma cell lines, U9 and HD3, which activate approximately equal amounts of TGF-beta 1 and express equal amounts of TGF-beta receptors, are now used to compare the effects of TGF-beta 1 in modulating invasive behavior. The U9 cell line exhibits autocrine-positive growth regulation in vitro by TGF-beta 1, whereas the HD3 cell line shows the opposite response, autocrine-negative regulation. Blocking endogenous TGF-beta 1 with isotype-specific antibody inhibited U9 cell growth because autocrine TGF-beta 1 acts as a mitogen for U9 cells. In contrast, antibody to TGF-beta 1 stimulated HD3 cell proliferation because autocrine TGF-beta 1 inhibits growth of these cells. U9 cells were 13-fold more invasive in vitro through a collagen I layer than HD3 cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

Molecular cloning and structure of the human (GABATHG) GABA transporter gene.

A cDNA molecule encoding the human GABA transporter was synthesized by means of polymerase chain reaction (PCR) technique and used as probe for selecting a human genomic DNA fragment encoding GABA transporter. A positive clone harboring the whole gene was obtained from a human lymphocyte genomic library through utilizing the genomic 'walking' technique. The clone, designated as pHGAT, harbours a DNA fragment of about 39 kb in length inserted into the BamHI site in cosmid pWE15. The gene covers about 25 kb in length and is constituted by four EcoRI restricted fragments which are 13.7 kb, 3.1 kb, 4.2 kb and 7.2 kb long, respectively. The genomic clone contains 15 introns, including two introns prior to the initiator methionine (i.e., the translation start site is in exon 3). Eleven exons encode the twelve transmembrane regions in the transporter protein. Thus as in the case for a number of other membrane proteins, there appears to be a strong tendency for the putative transmembrane domains to be encoded by separate exons. It is noted that the structure of the human GABA transporter gene reported here differs from the mouse gene which is contains 12 introns.

Base Sequence↗
PubMed1993

Can the stability of protein mutants be predicted by free energy calculations?

The use of free energy simulation techniques in the study of protein stability is critically evaluated. Results from two simulations of the thermostability mutation Asn218 to Ser218 in Subtilisin are presented. It is shown that components of the free energy change can be highly sensitive to the computational details of the simulation leading to the conclusion that free energy calculations cannot currently be used to reliably predict protein stability. The different factors that undermine the reliability are discussed.

Asparagine↗
PubMed1993

Regulation of Na,K-ATPase beta 1 mRNA content by thyroid hormone in neonatal rat cardiac myocytes.

Incubation of primary cultures of neonatal rat cardiac myocytes in the presence of 100 nM triiodothyronine (T3) resulted in a three- to fivefold increase in the content of Na,K-ATPase beta 1 subunit mRNA which was maximal at 1 d of exposure to hormone. To investigate the mechanism by which T3 stimulates the abundance of beta 1 mRNA, transient transfection experiments were conducted with a chimeric gene containing a portion of the 5' end of the rat beta 1 gene linked to a luciferase reporter gene. We found no effect of T3 on chimeric gene activity either in the absence or presence of cotransfected T3 receptor. The effect of T3 on the transcription rate of the endogenous beta 1 gene was quantitated by the nuclear run-on assay. T3 had no effect on beta 1 gene transcription following either 1 or 3 d of exposure and yielded a 1.3-fold increase at 6 d. These data indicate that T3 induction of Na,K-ATPase beta 1 mRNA content in neonatal rat cardiac myocytes in vitro is primarily mediated at a post-transcriptional site.

Analysis of Variance↗
PubMed1992

Effect of glycolipids on the phase behavior and dynamic properties of phospholipid liposomes.

The glycolipids of Acholeplasma laidlawii AIH089 membranes were identified and purified. The effect of monoglucosyldiacylglycerol (MGDG) and diglucosyldiacylglycerol (DGDG) on the thermotropic behavior of multilamellar vesicles of dipalmitoylphosphatidylcholine (DPPC), dipalmitoylphosphatidylglycerol (DPPG) has been investigated by high sensitivity differential scanning calorimetry. The main transition peaks were broadened, the enthalpies were decreased. DGDG caused the decrease in the transition temperatures of DPPC, DPPG liposomes by 3.08 degrees C, 4.18 degrees C, respectively. MGDG did not cause the alteration of the transition temperature of DPPC liposomes but caused the decrease of the transition temperatures of DPPG liposomes by 2.20 degrees C. ESR experiments indicate that MGDG decreased the rotational correlation time of DPPC and DPPG liposomes.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed1992

Multiple transcripts encoded by the ilvGMEDA gene cluster of Escherichia coli K-12.

We report here that, using Northern (RNA) blots, we identified two relatively stable transcripts of 4.6 and 1.1 kb that correspond to the products of the ilvEDA and ilvE genes and two relatively unstable transcripts of 6.7 and 3.6 kb that correspond to the products of the ilvGMEDA and ilvDA genes. The transcripts were identified by the use of eight probes derived from segments of the ilvGMEDA cluster. In addition, we used two strains with deletions of ilvG or ilvDA and observed the expected decrease in transcript size in Northern blots. Primer extension with reverse transcriptase generated a 169-nucleotide product corresponding to a 5' end within the ilvED intercistronic region, 37 nucleotides from the AUG codon of the ilvD gene. This primer extension product presumably indicates the 5' end of the ilvDA transcript that we detected in Northern blots. The stability of the transcripts was monitored, and RNase E was found to play a major role in ilv transcript degradation. Transcript levels varied in response to growth in the presence of the end product amino acids and in response to the presence of the polar frameshift site in ilvG. Although there have been speculations about the identities and numbers of transcripts derived from the ilvGMEDA cluster on the basis of the identification of some of the sites of transcription initiation and termination, this is the first report of the use of Northern blots to determine the actual sizes and distribution of mRNAs present in vivo.

Escherichia coli↗
PubMed1992

Recombinant human G-CSF and retinoic acid in synergistically inducing granulocyte differentiation of human promyelocytic leukemic cells.

The effects of recombinant human G-CSF (rhG-CSF) and retinoic acid (RA) were studied on the proliferation and differentiation of HL-60 cells and human acute myeloid leukemic cells. Synergistic effect on granulocyte differentiation was observed when HL-60 cells and primary acute promyelocytic leukemic cells were cocultured with RA plus rhG-CSF. rhG-CSF combined with RA increased more significantly the percentage of mature cells than RA alone and greatly increased NBT reduction activity (P < 0.001). These results suggested that proliferated effect of rhG-CSF on leukemic cells may be important for inducing differentiation of myeloid leukemic cells. But this effect might expose the patients to the risk of acute myeloblastic leukemia if G-CSF was used alone. However, RA could not only rule out the latter situation but retain former merit as well. The authors suggest that the combined use of G-CSF with RA is probably a new approach to the treatment of leukemia.

Cell Differentiation↗
PubMed1992

[Role of enteric Klebsiella pneumonia infection and HLA-B27 in ankylosing spondylitis].

Klebsiella pneumonia (KP) infection and HLA-B 27 have been shown to be strongly associated with ankylosing spondylitis (AS). In the present study, faecal cultures were performed and showed faecal carriage rate of KP was much higher in patients with AS (10/30) and hospital volunteers (2/10) than in the non-hospital volunteers (0/20). An octadecapeptide encompassing the shared hexamer between HLA-B 27 and KP nitrogenase residue was synthesized and autoantibodies against this short peptide were detected in sera of patients with AS and Reiter's syndrome (RS) and other related disease and normal controls. The results showed that such autoantibodies were detected in 42.2% of AS and 30% of RS patients yielding positive rate much higher than those found in other control groups. It is concluded that enteric KP infection were strongly implicated in the pathogenesis of AS probably by the mechanism of molecular mimicry with HLA-B 27.

Amino Acid Sequence↗
PubMed1991

Sequence and transcriptional activity of the Escherichia coli K-12 chromosome region between rrnC and ilvGMEDA.

We previously identified a protein related to the expression of the ilvGMEDA cluster of Escherichia coli K-12. It was observed that this ilv-related protein was produced at higher levels in UV irradiated cells infected with lambda dilvGMEDA phage with specific ilvG mutations (ValR), compared to phage carrying the wild-type(ValS) ilvG allele. The gene encoding this protein was further localized to a region between rrnC and ilvGMEDA by analyzing restriction fragment subsets in maxicells. We have now determined the nucleotide (nt) sequence of the 3.5-kb segment between rrnC and ilvGMEDA, and two open reading frames (ORFs) are present in the region expected to contain the ilv-related gene. These ORFs predicts Mrs of 18,751 (ORFI) and 20,085 (ORFII) Da, and both ORFs have a strong probability to encode proteins based on codon frequency analysis. Maxicell analysis revealed that a 1319-bp HindIII-SmaI fragment containing ORFI encodes the ilv-related peptide. We deleted a ClaI fragment that removed a portion of ORFI encoding the C-terminal region of the peptide, and maxicell analysis revealed a decrease in the size of the protein produced in accord with the prediction. RNA slot blots and Northern blots were used to characterize transcripts encoding ORFI. A transcript initiated 112 nt from the ilvGp2 promoter, but proceeding in the opposite direction, may encode the ORFI peptide.

Amino Acid Sequence↗
PubMed1991

The emergence of sense organs in the wing disc of Drosophila.

We have examined the origin of a set of precisely located sense organs in the notum and wing of Drosophila, in transformant flies where lacZ is expressed in the progenitor cells of the sense organs (the sensory mother cells) and in their progeny. Here we describe the temporal pattern of appearance and divisions of the sensory mother cells that will form the eleven macrochaetes and the two trichoid sensilla of the notum, and five campaniform sensilla on the wing blade. The complete pattern of sensory mother cells develops in a strict sequence that extends over most of the third larval instar and the first 10 h after puparium formation. The delay between the onset of lacZ expression and the first differentiative division ranges from 30 h, in the case of the earliest mother cells, to 2 h for the latest mother cells. The first division shows a preferential orientation which is also specific for each sensory mother cell. Up to this stage, there is no marked difference between the three types of mechanosensory organs.

Animals↗
PubMed1991

Design and compliance of HBV vaccination trial on newborns to prevent hepatocellular carcinoma and 5-year results of its pilot study.

A large-scale, controlled study of universal immunization of newborns against HBV infection has been conducted in the high incidence area of hepatocellular carcinoma, Qidong County of China. This area has a stable population, standardized cancer registration system, and an epidemiological base for measurements of liver cancer prevention by vaccine. Randomization was done on the community level. The vaccination and the control group each will consist of 38,000 children by the end of 1990. It is anticipated that the design will provide high statistical power to detect 50% reduction in the prevalence rate of chronic hepatitis among the vaccinees vs. the controls at 6 to 10 years of age, and 50% reduction in the incidence rate of hepatocellular carcinoma at 35 to 40 years of age. The vaccine used is Hep-B Vax, donated by Merck and Co. through WHO. The vaccine was administered at 0, 1, and 6 months after birth, the dosage of 5 or 2.5 micrograms in the pilot study as used before 1985 and of 5 micrograms dose level during the main study starting from January 1, 1985. About 85% of the cohorts have now entered the protocol. The vaccination coverage during 1984 to 1989 was 98.0% (35,064/35,789). Follow-up of the vaccinees and the age-matched controls at 5 years has exceeded 97%. The cumulative mortality in the vaccinated group up to 1988 was 1.29% (354/27,450). No single death nor serious adverse reaction was found that was associated with vaccination. The use of HBV vaccine at a reduced dose was especially important for the developing countries at the present time in order to achieve widespread immunization. Five-year results of the pilot study of this vaccination project showed that significant protection against HBV infection was achieved with the 5 or 2.5 micrograms per dose regimen plus a booster of 5 micrograms given at 3.5 to 4 years of age.(ABSTRACT TRUNCATED AT 250 WORDS)

Carcinoma, Hepatocellular↗
PubMed1991

Regeneration of transgenic Lycium barbarum L.

A simple and effective system for the transformation and regeneration of Lycium barbarum L. has been developed. Young stem segments from Lycium barbarum L. were infected by Agrobacterium tumefaciens harboring a vector containing neomycin phosphotransferase II (npt-II) gene derived from non-oncogenic Ti plasmid. Calli originating from young stem segments on selective induction medium could differentiate into buds on selective differentiation medium rapidly and finally developed into whole plants. NPT-II enzyme activity assay and DNA hybridization indicated that the foreign gene had been integrated into the genome of Lycium barbarum L. and could be expressed in plants.

Kanamycin↗