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Biomedical subjects

C Li

Publications and source records attributed to C Li.

At least 1,099 records · Page 61Linked to original sources

A reevaluation of the critical membrane potential of the effective refractory period in guinea pig ventricular fibres.

The traditional critical membrane potential (CMP), -55(-)-60 mV, which corresponds to effective refractory period (ERP), was anew investigated in guinea pig ventricular muscle fibres. The electrical and contractile responses to the stimulus during repolarization of action potential (AP), particularly from +10 to -60 mV, were observed. One third of 35 tested cells displayed testing action potential (TAP) and local response at > or = -54 mV when they were stimulated by testing pulses in 37 degrees C normal Tyrode's solution. Potential level of TAP which occurred earliest was at -30 mV and that of local response which appeared earliest was at 0 mV during repolarization among 95 systematic tests. Most of the TAPs belonged to the slow response potential type. The ratio of TAP evoked at > or = -54 mV initial membrane potential (IMP) was as high as 86% when the experiment was carried out in 37 degrees C 1.5 mmol KC1/L Tyrode's solution. In view of distribution of IMPs of TAPs, the CMP of ERP in guinea pig ventricular muscle fibres was more positive than traditional CMP measured by Hoffman et al. in dog, sheep Purkinje fibres and had a quite changeable range. The CMP of every cell in ventricular muscle was not all the same, and their CMPs approximated to normal distribution. There was no sharp line separating ERP from relative refractory period in myocardium. Higher temperature and low [K]0 were the important factors elevating CMP of ERP.

Action Potentials↗

[Investigation of a Candida albicans-specific DNA fragment used for nucleic acid hybridization].

A 2.5 kb fragment from Candida albicans Cli EcoRI-digested was labeled with 32P and used to probe blots of DNA from various organisms. DNA from all strains of C. albicans gave evidence of strong sequence homology with the 2.5 kb fragment. With the exception of weak hybridization signals from strains of C. parapsilosis and a strain of C. tropicalis, DNA from other Candida species, Cryptococcus as well as bacteria did not hybridize with the probe. The results indicate that the 2.5 kb fragment might contain sequence unique to C. albicans. After this is confirmed by cloning, a specific probe for the identification of C. albicans will be made.

Blotting, Southern↗

Ethanol inhibits a neuronal ATP-gated ion channel.

The cellular mechanisms by which ethanol affects nervous system function are poorly understood. However, evidence has been accumulating that ethanol can affect the function of neurotransmitter-gated ion channels. Extracellular ATP has recently been reported to produce excitatory actions in the peripheral and central nervous systems by activating ligand-gated ion channels. We studied the effect of ethanol on membrane ion current activated by extracellular ATP in isolated bullfrog dorsal root ganglion neurons, by means of the whole-cell patch-clamp technique. The amplitude of the ATP-activated current was decreased by ethanol in a concentration-dependent manner over the range of 3-500 mM. The average inhibition of 1 microM ATP-activated current by 100 mM ethanol was 64 +/- 3%, and the concentration of ethanol that produced 50% inhibition was 68 mM. Ethanol inhibition of ATP-activated current was not dependent on membrane potential from -80 to +40 mV, and ethanol did not change the reversal potential of ATP-activated current. Ethanol (100 or 400 mM) shifted the ATP concentration-response curve to the right, increasing the EC50 for ATP from 3.0 microM to 6.0 microM or 22.3 microM, respectively, but did not reduce the maximal response to ATP. The results suggest that ethanol inhibits ATP-activated current by increasing the apparent dissociation constant for the ATP receptor.

Adenosine Triphosphate↗

[Study on degradation of phenols in water].

The degradation of phenols in water and the effects of microbiolism, pH and dissolved oxygen (DO) on the degradation were investigated. The results showed that the degradation of phenols in water was mainly that of biochemistry, which depends on the existence of microbiolism. The most suitable pH for the degradation was 6 to 9, and no effects of DO on it were found. The accustomization of microbiolism with phenols would accelerate the degradation.

Hydrogen-Ion Concentration↗

Recent advances in studies on traditional Chinese anti-aging materia medica.

Presented in this paper is a report of our studies on 386 traditional effective anti-aging medications, the effects of which on cell generation, survival time, immunomodulation, improvement of visceral and metabolic functions, and anti-infection, and their trace element contents were further summarized and analysed. This suggests that the investigations of traditional anti-aging materia medica in China are now well under way and some effective drugs and compound prescriptions have been explored, such as Ginseng, Radix Astragali seu Hedysari, Radix Angelicae Sinensis, Herba Epimedii, Cordyceps, Ganoderma Lucidum seu Japonicum, Radix Polygoni Multiflori, Radix Acanthopanacis Senticosi, Rhizoma Polygonati, Fructus Lycii, and Poria. However, all of these preliminary results remain to be further investigated.

Aging↗

5-Lipoxygenase activity in the human pancreas.

Low levels of 5-lipoxygenase (5-LO), the first committed enzyme in the synthesis of leukotrienes (LTs), have been reported in the porcine pancreas. We have quantitated 5-LO activity in subcellular fractions of pancreas samples from three human donors. 5-LO activity was detectable in all samples, although enzyme activity was lower than in human leukocytes. 5-LO in human pancreas samples displayed highest specific activity in membrane fractions, and did not require arachidonic acid (AA) addition for activity. These unusual characteristics of pancreatic 5-LO appear to be due, at least in part, to the presence of unesterified AA in the pancreas samples. Western blot analysis demonstrated that the human pancreas contains low levels of 5-lipoxygenase-activating protein (FLAP) in addition to 5-LO.

5-Lipoxygenase-Activating Proteins↗

[Preconcentration of trace Fe, Mn, Cu and Cd in water with charcoal and determination by flame atomic absorption spectroscopy].

The absorption of Fe, Mn, Cu and Cd by charcoal and the influence of pH, particle size and temperature on absorption were studied. A method for the preconcentration of these irons in water was established, and with flame atomic absorption spectroscopy a content as low as microgram/L could be determined. The recovery and the coefficient of variance (n = 6) were in the range of 90-113% and 2.6-9.9%, respectively. The proposed method is simple, rapid, accurate reproducible and suitable for the preconcentration and determination of titled metal irons in different kinds of water.

Cadmium↗

Follow-up of asymptomatic myocardial ischemia in patients with diabetes mellitus.

Early detection of asymptomatic myocardial ischemia in patients with diabetes mellitus (DM) is very important for the prevention of painless myocardial infarction. The purpose of this study is to explore the clinical significance of asymptomatic transient ST changes in ECG in patients with DM and to further clarify the relation between asymptomatic myocardial ischemia and myocardial infarction in DM.

Aged↗

Hemodynamic and nonhemodynamic mechanisms of experimental pulmonary edema in rats and the effect of anisodamine and tetramethylpyrazine. Part 1: Survival rate, pulmonary index, pathological change and pulmonary vascular permeability.

Pulmonary edema (PE) which is similar to the neurogenic type was induced by adrenaline (AD) administration (0.1 mg/kg) in rats. Acute progressive respiratory distress, cyanosis and dyspnea occurred. All the experimental animals in the PE group died within 20 min after AD injection, with a pulmonary index (PI) of 1.70 +/- 0.47 (mean +/- S) which was much higher than that in the normal group. The mortality rate was 100%. It was found that in rats with PE, a protein-rich fluid filled the alveolar and interstitial spaces, and ecchymosis occurred. The capillary permeability as estimated by Evans blue injection showed that Evans blue from extraction fluid and bronchoalveolar lavage (BAL) in the PE rats was at a much higher level than that in the normal control (NC) rats. In anisodamine (ADM, 654-2) and tetramethylpyrazine (TMP) treated rats, almost all the damage was diminished or absent, and the mortality rates were decreased from 100% to 4.4% and 20%, respectively. 654-2 and TMP could significantly inhibit the increase of pulmonary permeability.

Animals↗

Ultrastructural study of moniliformin induced lesions of myocardium in rats and mice.

Effects of moniliformin on the ultrastructure of the myocardium of mice and rats were studied. Mice were given moniliformin orally at a dose of 29.46 mg.kg-1 the LD50. One h after dosing, lesions of the mitochondria of the myocardial cells were found which became more severe in 2 and 3 h. Ultrastructural lesions were also observed in the myofibrils and sarcolemma. Rats were given moniliformin orally at the dosage of 6 mg.kg-1 once daily for 56 d. Lesions of mitochondria and myofibrils were relatively mild. In the myocardiac specimens taken from the 21d post-toxin administration, lesions of the sarcolemma became more obvious. These moniliformin-induced lesions were similar to the ultrastructural changes in the myocardium of patients with Keshan disease. Our findings indicate that there may be a close and important relationship between moniliformin intoxication and Keshan disease.

Animals↗

Targeting of Sp1 to a non-Sp1 site in the human neurofilament (H) promoter via an intermediary DNA-binding protein.

The human neurofilament (H) promoter contains multiple binding sites for nuclear proteins including a Proximal (Prox) site centered around the sequence GGTTGGACC and an adjacent pyrimidine (Pyr) tract site centered around the sequence CCCTCCTCCCC. Surprisingly binding to a probe containing the Prox/Pyr region of the NF(H) promoter was competed in gel shifts by an oligonucleotide containing only an Sp1 binding site (GGGGCGGGG). Supershift assays with a polyclonal anti-Sp1 antisera confirmed that Sp1 was part of the complex formed with the Prox/Pyr probe. However neither bacterially expressed Sp1 516C or vaccinia virus expressed full-length Sp1 778C bound to the Prox or Pyr sequences in DNase I footprints or gel shift assays. Gel shift competitions and supershift assays with probes containing either Prox or Pyr tract sites alone demonstrated targeting of Sp1 to the Prox binding site and identified a non-Sp1 containing complex which contains a Prox binding protein. Adding exogenous Sp1 to a HeLa nuclear extract enhanced the Sp1-containing complex but had no effect on the Prox complex. These studies show that Sp1 can be targeted to a non-Sp1 site in the human NF(H) promoter through protein/protein interactions with a distinct sequence specific DNA-binding protein.

Base Sequence↗

A protein methyltransferase specific for altered aspartyl residues is important in Escherichia coli stationary-phase survival and heat-shock resistance.

Proteins are subject to spontaneous degradation reactions including the deamidation, isomerization, and racemization of asparaginyl and aspartyl residues. A major product of these reactions, the L-isoaspartyl residue, is recognized with high affinity by the protein-L-isoaspartate(D-aspartate) O-methyltransferase (EC 2.1.1.77). This enzyme catalyzes the methyl esterification of the L-isoaspartyl residue in a reaction that can initiate its conversion to the normal aspartyl configuration. To directly study the physiological role of this methyltransferase, especially with respect to the potential repair of isomerized aspartyl residues in aging proteins, we examined the ability of the bacterium Escherichia coli to survive in the absence of its activity. We utilized gene disruption techniques to replace the chromosomal copy of the pcm gene that encodes the methyltransferase with a kanamycin-resistance cassette to produce mutants that have no detectable L-isoaspartyl methyltransferase activity. Although no changes in exponential-phase growth were observed, pcm- mutants did not survive well upon extended culture into stationary phase or upon heat challenge at 55 degrees C. These results provide genetic evidence for a role of the L-isoaspartyl methyltransferase in the metabolism of altered proteins that can accumulate in aging cells and limit their viability.

Aspartic Acid↗

Monoclonal antiprothrombinase (3D4.3) prevents mortality from murine hepatitis virus (MHV-3) infection.

The induction of monocyte/macrophage procoagulant activity (PCA) has been implicated in the pathogenesis of murine hepatitis virus strain 3 (MHV-3) infection and disease. Previously, we have shown that induction of PCA by MHV-3 correlated with resistance/susceptibility to infection in different mouse strains. In this study, all BALB/cJ mice that were infected with 10(3) plaque-forming units of MHV-3 developed severe liver disease and died within 96-120 h. Examination of the livers of these animals showed marked hepatic necrosis, deposition of fibrin, and cellular expression of PCA by direct immunofluorescence staining in areas of necrosis as well as in hepatic sinusoids. Splenic mononuclear cells recovered from these mice expressed high concentrations of PCA with time after infection. Infusion into mice of a high-titered monoclonal antibody that neutralized PCA (3D4.3) attenuated the development of hepatic necrosis and enhanced survival in a dose-dependent manner. All of the animals receiving 100 micrograms, and 44% and 22% of the animals that received 50 and 25 micrograms per day, respectively, survived for 10 d and made a full recovery. Administration of the antibody resulted in a dose-dependent reduction in fibrin deposition, PCA expression as detected by direct immunofluorescence staining and by a functional assay. In animals treated with high concentrations of antibody, titers of antibody to PCA fell from 87 +/- 15 micrograms/ml to 100 +/- 7 ng/ml during the active phase of the disease, consistent with sequestration due to binding of the immunoglobulin to cells expressing PCA. Surviving animals, when rechallenged with MHV-3, had a 40% mortality, consistent with the known rates of metabolism of immunoglobulin. This further suggested that protection was by a passive mechanism. The results reported here demonstrate that a neutralizing antibody to PCA protects animals from fulminant hepatitis and death associated with MHV-3 infection, and supports the notion that PCA is a potent inflammatory mediator that plays a pivotal role in the pathogenesis of liver injury resulting from MHV-3 infection.

Animals↗

Molecular cloning and characterization of an adenosine receptor: the A3 adenosine receptor.

We have previously reported the selective amplification of several rat striatal cDNA sequences that encode guanine nucleotide-binding regulatory protein (G protein)-coupled receptors. One of these sequences (R226) exhibited high sequence identity (58%) with the two previously cloned adenosine receptors. A full-length cDNA clone for R226 has been isolated from a rat brain cDNA library. The cDNA clone encodes a protein of 320 amino acids that can be organized into seven transmembrane stretches. R226 has been expressed in COS-7 and CHO cells and membranes from the transfected cells were screened with adenosine receptor radioligands. R226 could bind the nonselective adenosine agonist tritiated N-ethyladenosine 5'-uronic acid ([3H]NECA) and A1-selective agonist radioiodinated N6-2-(4-amino-3-iodophenyl)-ethyladenosine ([125I]APNEA) but not A1-selective antagonists tritiated 1,3-dipropyl-8-cyclopentylxanthine ([3H]DPCPX) and 8-(4-[([[(2-aminoethyl)amino]carbonyl]methyl)oxy]-phenyl)-1, 3-dipropylxanthine ([3H]XAC) or the A2-selective agonist ligands tritiated 2-[4-(2-carboxyethyl)phenyl]ethyl-amino 5'-N-ethylcarboxamidoadenosine ([3H]CGS21680) and radioiodinated 2-[4-([2-[(4-aminophenyl)methylcarbonylamino] ethylaminocarbonyl]ethyl)phenyl]ethylamino 5'-N-ethylcarboxamidoadenosine. Extensive characterization with [125I]APNEA showed that R226 binds [125I]APNEA with high affinity (Kd = 15.5 +/- 2.4 nM) and the specific [125I]APNEA binding could be inhibited by adenosine ligands with a potency order of (R)-N6-phenyl-2-propyladenosine (R-PIA) = NECA greater than S-PIA greater than adenosine greater than ATP = ADP but not by antagonists XAC, isobutylmethylxanthine, and DPCPX. In R226 stably transfected CHO cells, adenosine agonists R-PIA, NECA, and CGS21680 inhibited by 40-50% the forskolin-stimulated cAMP accumulation through a pertussis toxin-sensitive G protein with an EC50 of 18 +/- 5.6 nM, 23 +/- 3.5 nM, and 144 +/- 34 nM, respectively. Based on these observations we conclude that R226 encodes an adenosine receptor with non-A1 and non-A2 specificity, and we thus name it the A3 adenosine receptor. mRNA analyses revealed that the highest expression of R226 was in the testis and low-level mRNAs were also found in the lung, kidneys, heart, and some parts of the central nervous system such as cortex, striatum, and olfactory bulb. The high-expression level of the A3 receptor in the testis suggests a possible role for adenosine in reproduction.

Adenosine↗

1,9-Dideoxyforskolin does not mimic all cAMP and protein kinase A independent effects of forskolin on GABA activated ion currents in adult rat sensory neurons.

The effect of forskolin on GABAA receptor activated events has been the subject of recent investigations, the conclusions of which are conflicting. Forskolin can reduce current amplitude and increase the rate of decay of current activated by 100 microM GABA and these effects are not mimicked by 1,9-dideoxyforskolin (Tehrani et al., Synapse, 4 (1989) 126-131). On the other hand, both forskolin and 1,9-dideoxyforskolin inhibit 36Cl- flux induced by lower concentrations of muscimol (Heuschneider and Schwartz, Proc. Natl. Acad. Sci. USA, 86 (1989) 2938-2942). Using the whole-cell patch clamp technique to measure GABA activated current in dorsal root ganglion neurons that were freshly isolated from adult rats, we have confirmed the finding of Tehrani et al. (Synapse, 4 (1989) 126-131) using 100 microM GABA; however, the effects of forskolin that were not mimicked by 1,9-dideoxyforskolin were not blocked by the kinase inhibitor H-7 (50 microM). In contrast, at lower concentrations of GABA (10-20 microM), both forskolin and 1,9-dideoxyforskolin increased the decay rate of GABA activated current. In addition, all effects of forskolin occurred within 200 ms of application of forskolin and the effects were not blocked or occluded by H-7, 10 microM cAMP, or the active subunit of protein kinase A. We conclude that: (1) 1,9-dideoxyforskolin is not a reliable indicator of forskolin specificity in this system because its effects are dependent upon GABA concentration; and (2) the most prominent effects of forskolin on amplitude and decay time course of GABA activated ion current are not mediated by cAMP or protein kinase A (PKA).

Animals↗

Peptide amidating enzymes are present in cultured endothelial cells.

Carboxy-terminal amidation is a prevalent post-translational modification necessary for the bioactivity of many peptides. We now report that the two enzymes essential for amidation, peptidylglycine alpha-monooxygenase (PAM) and peptidylamidoglycolate lyase (PGL), are present in both the cytosol and membrane fractions of cultured bovine aortic endothelial cells. Endothelial PAM exhibits ascorbate-dependent turnover and is inactivated by the mechanism-based inactivator, 4-phenyl-3-butenoic acid (PBA), whereas PGL activity is independent of ascorbate and is not affected by PBA. These enzymological characteristics correspond to those of amidating enzymes from other tissues. These results suggest a heretofore unrecognized role for alpha-amidated peptides in cardiovascular function.

Amidine-Lyases↗

Localization of FMRFamide-like peptides in Caenorhabditis elegans.

The neuropeptide FMRFamide (Phe-Met-Arg-Phe-NH2) is a member of a large family of related peptides that have been found throughout the animal kingdom. By using an antiserum specific for the Arg-Phe-NH2 moiety, we have found that about 10% of the neurons in the nematode Caenorhabditis elegans are immunoreactive. Most of these neurons, which include sensory, motor, and interneurons, were identified on the basis of their number, position, and projection pattern and by analysis of characterized mutants. Neurons that were immunoreactive in hermaphrodite animals were generally also found in males, but each sex had, in addition, sex-specific immunoreactive cells. Staining of hermaphrodite animals from different larval stages suggests that the onset of FMRFamide-like expression is differentially regulated among the cells. We have found a possible neuromodulatory role for the related peptide FLRFamide (Phe-Leu-Arg-Phe-NH2). In an egg-laying assay, FLRFamide by itself was not active but could potentiate a serotonin effect. The FMRFamide-like immunoreactivity was also used as a marker to examine the differentiation of cells that normally undergo programmed cell death. Cells that are destined to die in the Pn.a lineages appear to differentiate and adopt the fate of lineally equivalent cells before cell death.

Amino Acid Sequence↗