External use of bi yuan ning in 200 cases of nasosinusitis and rhinitis.
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Administration, Intranasal↗
Biomedical subjects
C Huang
Publications and source records attributed to C Huang.
[Analysis of surface antigen molecule expression on serovar stains of Neisseria gonorrhoeae by flow cytometry].
In the present experiment, flow cytometry was employed for analysing expression characteristics of the antigen molecules distingushed by 10 monoclonal antibodies against Neisseria gonorrhoeae lipooligosaccharides. Stability of antigen expression and amount of epitope on the surface of N. gonorrhoeae were quantitative determined. Reactivity of the monoclonal antibodies with serovar strains of N. gonorrhoeae were evaluated.
Antibodies, Monoclonal↗
Effects of tyrphostins on activity of casein kinase II from rat liver.
AIM: To investigate the effects of tyrphostins, (AG213, AG1394, AG114, AG1109, AG555) on the activity of casein kinase (CK) II. METHODS: CK II was partially purified from rat livers by sequential DE52 and heparin-Sepharose chromatography. CK II activity was assayed by incubating CK II with dephosphorylated casein and [gamma-32 P]ATP. RESULTS: AG213 inhibited the activity of CK II with IC50 44.7 mumol.L-1 (41.5-47.9 mumol.L-1), and AG1394 (144 mumol.L-1) strongly inhibited the activity of CK II with an inhibitory ratio of 89%. AG114 (174 mumol.L-1) and AG1109 (126 mumol.L-1) had inhibitory effects on the activity of CK II (p < 0.01). AG555 (136 mumol.L-1) had little effect on CK II activity. CONCLUSION: Some tyrphostins are potent inhibitors of CK II.
Animals↗
Ripple phase in asymmetric unilamellar bilayers with saturated and unsaturated phospholipids.
In a solution of phosphate-buffered saline (PBS), unilamellar bilayers with saturated phosphatidylcholines in one leaflet and negatively charged, unsaturated phospholipids in the other leaflet were observed in the ripple phase at room temperature using atomic force microscopy (AFM). This is the first observation of the ripple phase in asymmetric bilayers. Sodium and phosphate, components of PBS, were found to be necessary for the formation of the ripple structure in the asymmetric bilayers composed of dipalmitoylphosphatidylcholine (DPPC) and 1-palmitoyl-2-oleoylphosphatidylglycerol (POPG), demonstrating a dependency for specific ions for this phase. These results indicate that the two leaflets of a bilayer are closely coupled to give rise to such a long range and complicated morphology.
Ions↗
Effect of nitrendipine on renal cortical and papillary autoregulation in hypertensive rats.
Renal cortical and papillary perfusions were assessed by laser-Doppler flowmetry and autoregulatory indices. There was cortical autoregulation (autoregulatory index of 0.31 +/- 0.08) in Wistar from 160 to 100 mm Hg, but not in the papilla, which was abolished by nitrendipine, 0.125 and 0.25 micrograms kg-1 min-1. In SHRSP the cortex, but not the papilla, exhibited autoregulation from 180 to 120 mm Hg (autoregulatory index of 0.27 +/- 0.10) but not during low and high doses of nitrendipine. The non-clipped kidney cortex of 2K1C Goldblatt rats autoregulated from 190 to 130 mm Hg (autoregulatory index of 0.20 +/- 0.13), but was inhibited only by the higher dose of nitrendipine. In mature Wistar and SHRSP cortical autoregulation is blocked effectively by nitrendipine whereas the 2K1C Goldblatt hypertensive rats are relatively resistant.
Analysis of Variance↗
Phospholipase D hydrolyzes ether- and ester-linked glycerophospholipids by different pathways in MDCK cells.
MDCK cells were prelabeled with 1-O-[3H]hexadecyl-2-lyso-GPC and [14C]myristic acid, which selectively labeled the glycerophospholipid subclasses with 93% of tritium in the alkyl-linked subclass and 85% of carbon-14 in the diacyl-linked subclass. By this approach, we have demonstrated that PLD upon activation via PKC pathway selectively catalyzes the degradation of ether-linked glycerophospholipid subclass. In contrast, G-protein regulatory PLD activity seems to preferentially hydrolyze ester-linked subclass. These results suggest that the selective hydrolysis of PLD action may play an important role in cellular signal transduction under physiological and pathological conditions.
Animals↗
A binding interface on the I domain of lymphocyte function-associated antigen-1 (LFA-1) required for specific interaction with intercellular adhesion molecule 1 (ICAM-1).
Previous studies have shown that lymphocyte function-associated antigen-1 (LFA-1) molecules containing the human alpha (CD11a) and human beta (CD18) subunits but not the murine alpha and human beta subunits can bind to human intercellular adhesion molecule 1 (ICAM-1). Using human/mouse LFA-1 alpha subunit chimeras, we mapped regions required for binding to ICAM-1 N-terminal to amino acid (aa) residue 350. Ligand binding sites were mapped in greater detail by scanning this region with murine sequences from 56 down to 17 aa in length and finally by introducing single or few murine aa residue replacements into the human sequence. Replacement of two non-contiguous regions of aa residues 119-153 and 218-248 in the me domain with the corresponding mouse sequences abolished most binding to human ICAM-1, without affecting alpha beta subunit association or expression on the surface of transfected COS cells. Specific residues within the I domain found to be important were Met-140, Glu-146, Thr-243, and Ser-245. Using the recently solved structure of the Mac-1 (CD11b) I domain as a model (Lee, J.-O., Rieu, P., Arnaout, M.A., and Liddington, R. (1995) Cell 80, 631-638), these residues are shown to be located on the surface of the I domain surrounding the site to which Mg2+ is chelated, and fine a ligand binding interface. Mapping of the epitopes of a panel of mouse anti-human and rat anti-mouse monoclonal antibodies gave concordant results. Epitopes were mapped to two different regions in the N-terminal domain, four regions within the I domain, and two regions between the I domain and the EF hand-like repeats. Monoclonal antibodies to epitopes within the mid- to C-terminal portion of the I domain and the N-terminal portion of the region between the I domain and the EF hand-like repeats gave good inhibition of LFA-1-dependent homotypic aggregation with cells that express either ICAM-1 or ICAM-3 as the major LFA-1 ligand.
Amino Acid Sequence↗
The chloroplast gene encoding ribosomal protein S4 in Chlamydomonas reinhardtii spans an inverted repeat--unique sequence junction and can be mutated to suppress a streptomycin dependence mutation in ribosomal protein S12.
The ribosomal protein gene rps4 was cloned and sequenced from the chloroplast genome of Chlamydomonas reinhardtii. The N-terminal 213 amino acid residues of the S4 protein are encoded in the single-copy region (SCR) of the genome, while the C-terminal 44 amino acid residues are encoded in the inverted repeat (IR). The deduced 257 amino acid sequence of C. reinhardtii S4 is considerably longer (by 51-59 residues) than S4 proteins of other photosynthetic species and Escherichia coli, due to the presence of two internal insertions and a C-terminal extension. A short conserved C-terminal motif found in all other S4 proteins examined is missing from the C. reinhardtii protein. In E. coli, mutations in the S4 protein suppress the streptomycin-dependent (sd) phenotype of mutations in the S12 protein. Because we have been unable to identify similar S4 mutations among suppressors of an sd mutation in C. reinhardtii S12 obtained using UV mutagenesis, we made site-directed mutations [Arg68 (CGT) to Leu (CTG and CTT)] in the wild-type rps4 gene equivalent to an E. coli Gln53 to Leu ribosomal ambiguity mutation (ram), which suppresses the sd phenotype and decreases translational accuracy. These mutants were tested for their ability to transform the sd S12 mutation of C. reinhardtii to streptomycin independence. The streptomycin-independent isolates obtained by biolistic transformation all possessed the original sd mutation in rps12, but none had the expected donor Leu68 mutations in rps4. Instead, six of 15 contained a Gln73 (CAA) to Pro (CCA) mutation five amino acids downstream from the predicted mutant codon, irrespective of rps4 donor DNA. Two others contained six- and ten-amino acid, in-frame insertions at S4 positions 90 and 92 that appear to have been induced by the biolistic process itself. Eight streptomycin-independent isolates analyzed had wild-type rps4 genes and may possess mutations identical to previously isolated suppressors of sd that define at least two additional chloroplast loci. Cloned rps4 genes from streptomycin-independent isolates containing the Gln73 to Pro mutation and the 6-amino acid insertion in r-protein S4 transform the sd strain to streptomycin independence.
Amino Acid Sequence↗
Expression and site-directed mutagenesis of mouse prostaglandin E2 receptor EP3 subtype in insect cells.
A cDNA encoding for mouse prostaglandin E2 (PGE2) receptor EP3 subtype was cloned from a mouse kidney cDNA library by PCR using terminal primers derived from the known sequence of mouse lung EP3 receptor cDNA. The cloned cDNA was confirmed by sequencing and was expressed in Trichoplusia ni (MG1) insect cells using a baculovirus expression system. A specific protein of 60 kDa was detected by immunoblot with antibodies generated against a unique decapeptide sequence present in the second extracellular loop of the EP3 receptor. Specific binding of [3H]PGE2 with a Kd of 3 nM was also found in the membrane fraction of the insect cells. Ligand binding of the receptor was further studied by site-directed mutagenesis. Arg-309 of the receptor was separately mutated to lysine, glutamate and valine. cDNAs of the wild-type and mutant EP3 receptors were respectively expressed and studied in MG1 insect cells. Binding studies indicated that both glutamate and valine mutant EP3 receptors had no binding of [3H]PGE2. On the contrary, the lysine mutant receptor exhibited an even tighter binding (Kd = 1.3 nM) than the wild-type EP3 receptor. Immunoblot studies indicated that these receptors were expressed in a comparable amount in MG1 insect cells. These results suggest that Arg-309 of EP3 receptor may be essential in ligand binding through ionic interaction.
Amino Acid Sequence↗
Repression of protein kinase C and stimulation of cyclic AMP response elements by fumonisin, a fungal encoded toxin which is a carcinogen.
Fusarium moniliforme (FM) is a major fungal pathogen of corn and is involved with stalk rot disease. FM is widely spread throughout the world, including the United States. Most strains of FM produce several mycotoxins, the most prominent of which is called fumonisin. Recent epidemiological studies indicated that ingestion of fumonisin correlates with a higher incidence of esophageal cancer in Southern and Northern Africa and China. Furthermore, fumonisin causes a neurodegenerative disease in horses, induces hepatic cancer in rats, and induces pulmonary edema in swine. Considering that high levels of fumonisin have been detected in healthy and diseased corn grown in the United States, fumonisin may pose a health threat to humans and livestock animals. Structurally, fumonisin resembles sphingolipids which are present in the membranes of animal and plant cells. At the present time, very little is known concerning the mechanism by which fumonisin elicits its carcinogenic effect. Our studies indicate that fumonisin represses expression of protein kinase C and AP-1-dependent transcription. In contrast, fumonisin stimulated a simple promoter containing a single cyclic AMP response element. Since fumonisin did not alter protein kinase A activity, it appears that cyclic AMP response element activation was independent of protein kinase A. It is hypothesized that the ability of fumonisin to alter signal transduction pathways plays a role in carcinogenesis.
Animals↗
Detection of California serogroup viruses using universal primers and reverse transcription-polymerase chain reaction.
Universal primers have been identified and a protocol developed that are suitable for rapid detection of California encephalitis (CE) complex viruses in a reverse transcription-polymerase chain reaction (RT-PCR) assay. These primers correspond to sequences in the coding regions of the G2 glycoprotein of the middle-size RNA segment. The identities of the amplified products were confirmed by sequencing on the clones or PCR products. The technique is capable of detecting 40 plaque-forming units (PFU) directly on an ethidium bromide-stained agarose gel and the sensitivity increases to 0.4-1 PFU when a radiolabeled probe is used as the detector.
Animals↗
Comparison of the M RNA genome segments of two human isolates of La Crosse virus.
The M genomic RNA segments of La Crosse (LAC) virus isolates from the brains of two children autopsied 18 years apart in Wisconsin were molecularly cloned using a reverse transcriptase-PCR assay and the nucleotide sequences of the cDNAs determined. The M RNA of each virus contains 4526 nucleotides, similar to that reported previously for a New York mosquito isolate of LAC. There were 20 nucleotide differences between the two human isolates, which results in the prediction of 7 amino acid changes in the proteins encoded in the single, long open reading frame of the M segment. One of these predicted differences occurs in the G2 glycoprotein and six in the G1 glycoprotein. The two viruses were identical in terms of predicted amino acid sequence in the region believed to represent a nonstructural protein. These data have been further compared to those available for two other California serogroup isolates.
Amino Acid Sequence↗
A fluorimetric method for the detection of copper-mediated hydroxyl free radicals in the immediate proximity of DNA.
An optical method to detect copper-mediated hydroxyl free radicals generated close to DNA and other biomolecules has been developed. Low-molecular-weight polylysines were labeled with SECCA, a derivative of coumarin that generates the fluorescent 7-OH-SECCA following its interaction with hydroxyl free radicals in aqueous solution. These polylysines were then complexed with DNA to place the detector molecule SECCA in the vicinity of the nucleic acid. Following addition of copper sulfate (0-10 mumol dm-3), free radicals were generated by incubation with ascorbic acid (0-1 mmol dm-3) and hydrogen peroxide (0-1 mmol dm-3). A rapid increase in the induced fluorescence was observed corresponding to the formation of the fluorescent 7-OH-SECCA in the polylysine-nucleic acid complex. This fluorescence was not decreased significantly by addition of high concentrations of hydroxyl free-radical scavengers (DMSO, methanol, ethanol and tert-butanol), but was diminished by addition of relatively low concentrations of EDTA (0.1 mmol dm-3), histidine (0.1 mmol dm-3) or catalase (8.3 x 10(-5) mmol dm-3). On the other hand, when such reaction mixtures were incubated with SECCA molecules that were free in solution or SECCA-labeled polylysine in the absence of DNA, the induced fluorescence was diminished by all hydroxyl free-radical scavengers. The efficiency by which the scavengers reduce the fluorescence increases as their hydroxyl rate constant increases. The data indicate that the detector molecule SECCA can be used to detect copper-mediated hydroxyl free radicals generated close to DNA.
Animals↗
Regulation of ferredoxin gene in steroidogenic and nonsteroidogenic cells.
Ferredoxin is an electron transport intermediate for all the mitochondrial cytochromes P450. It is especially abundant in steroidogenic organs where it functions in steroid biosynthesis. The regulation of ferredoxin gene expression was studied in both steroidogenic and nonsteroidogenic cell lines. In steroidogenic cell line Y1, the expression of ferredoxin was stimulated by cAMP and repressed slightly by angiotensin II and phorbol ester PMA. These drugs exhibited the same effect on the basal promoter of the ferredoxin gene, which includes one TATA box and an SP1 site. In human adrenocortical cell line H295, the stimulation of the ferredoxin gene by cAMP was blocked by cycloheximide, as observed in bovine adrenocortical cell culture. In nonsteroidogenic cell lines such as HeLa and COS-1, the stimulation of ferredoxin gene expression by cAMP was not observed, although basal expression was strong. Transfection studies showed that the ferredoxin promoter could not be stimulated by cAMP in nonsteroidogenic cells. Therefore the steroidogenic cell-specific regulation and the general expression pattern appears to be a property unique to the ferredoxin gene.
Adrenal Cortex↗
Vertebral dimension measurements improve prediction of vertebral fracture incidence.
We investigated the independent contributions of vertebral size and shape, bone density, and existing fractures for predicting the risk of new vertebral fractures among postmenopausal Japanese-American women in Hawaii. Baseline measurements of bone density at the distal and proximal radius and the calcaneus were obtained in 1981, and at the lumbar spine in 1984. Incident fractures were documented on serial radiographs during an average of 8.0 years of follow up of 840 women. A positive difference of 1 standard deviation in vertebral depth increased the risk of new vertebral fractures by approximately 1.3-fold (95% confidence interval = 1.03, 1.66) after controlling for bone density and prevalent fractures. A combination of large vertebral depth (above the 66th percentile) and low bone density (below the 33rd percentile) increased fracture risk approximately six times compared to women with high bone density (above the 66th percentile) and small vertebral depth (below the 33rd percentile). We conclude that combining information about vertebral depth, bone density, and prevalent fracture appears to be better for predicting new fractures than any single variable alone.
Absorptiometry, Photon↗
Predicting vertebral deformity using bone densitometry at various skeletal sites and calcaneus ultrasound.
We investigated the usefulness of bone density measurements from multiple skeletal sites and calcaneus ultrasound for evaluating the probability of vertebral deformation. Bone mineral density (BMD) was measured at the second metacarpal and middle phalanges using radiographic absorptiometry of hand radiographs, and at the lumbar spine using dual-energy x-ray absorptiometry. Distal radius and proximal radius were measured using single-energy x-ray absorptiometry (SXA), expressed as bone mineral content (BMC, grams per centimeter), and as BMD (grams per square centimeter). The calcaneus was measured using both SXA (BMD) and broadband ultrasound attenuation (BUA). Among the women in this study (mean age 74, SD = 5), 84 women developed new vertebral deformations (57 cases with one and 27 cases with two or more deformations), which were identified on serial radiographs during an average of 9 years prior to the measurements of bone density. Logistic regression analysis was used to calculate odds ratios for risk of deformation corresponding to a 1-SD difference in density or ultrasound, adjusted for age. All bone measurements were significantly associated with vertebral deformation, with odds ratios (95% confidence intervals) ranging from 1.40 (1.10, 1.78) for proximal radius BMD to 1.88 (1.45, 2.44) for calcaneus BMD measurements. Measurements of calcaneal BUA, calcaneal BMD, and hand BMD generally remained significant when included simultaneously with another measurement in the same model, suggesting that spine or radius BMD may not provide much additional information about risk of deformation. It appears that all of the measurements of bone density and ultrasound provide useful information regarding the probability of deformation. These findings await confirmation in a prospective study.
Absorptiometry, Photon↗
Vertebral fracture prevalence in women in Hiroshima compared to Caucasians or Japanese in the US.
BACKGROUND: Although vertebral fractures are very common among elderly Caucasian women, no studies have compared the prevalence to that among Asian populations. Any observed differences in prevalence might lead to the identification of important environmental and/or genetic factors. We therefore compared the prevalence of vertebral fractures among US Caucasians to native Japanese and Japanese immigrants in Hawaii using a standardized approach. METHODS: Spinal radiographs of women aged > 50 years were obtained from native Japanese in Hiroshima, Japanese-Americans in Hawaii, and North American Caucasians in Minnesota between 1982 and 1991. Fractures were defined as vertebral heights > 3 standard deviations (SD) below the vertebra-specific mean. RESULTS: Compared to Japanese-Americans, odds ratios (OR) and 95% confidence intervals (CI) for prevalent vertebral fractures were 1.8 (95% CI: 1.3-2.5) for native Japanese women and 1.5 (95% CI: 1.1-2.1) for Minnesota Caucasians. The OR tended to be higher when comparing the prevalence of two or more fractures per person: OR = 3.2 (95% CI: 2.0-5.3) for native Japanese and OR = 1.9 (95% CI: 1.2-3.2) for Minnesota Caucasians. Similar results were observed for native Japanese using a fracture definition of > or = 4 SD below the mean, but the OR for Caucasians was reduced to 1.2 (95% CI: 0.6-2.3). CONCLUSION: The observation that, among these three populations, hip fracture incidence is lowest but spine fracture prevalence is greatest among native Japanese suggests that different risk factors may be responsible.
Age Factors↗
Improved safety from plasma derivatives: purification and viral elimination characteristics of mononine.
The occurrence of viral transmission with coagulation factor concentrates treated with a single virus elimination step has resulted in a consensus that multiple virus elimination steps must be incorporated in the processing of these proteins. As an example of this methodology, the steps included in the purification of the factor VIII concentrate Monoclate-P are reviewed and those used to purify the factor IX concentrate Mononine--monoclonal antibody chromatography, sodium thiocyanate incubation, and a novel ultrafiltration system--are described. Rigorous quality control/quality assurance during processing is essential. The ultimate safety of the product used clinically further requires monitoring of long-term stability and verification of the absence of neoantigens that could stimulate unusual levels of antibody formation. Standardization of regulatory compliance will further strengthen the safety of all products. The zeal to add further barriers to viral contamination should not compromise the timely assessment of these additional protections.
Antibodies, Monoclonal↗