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Biomedical subjects

J Xing

Publications and source records attributed to J Xing.

At least 55 records · Page 3Linked to original sources
PubMed1998

Nerve growth factor activates extracellular signal-regulated kinase and p38 mitogen-activated protein kinase pathways to stimulate CREB serine 133 phosphorylation.

The mechanisms by which growth factor-induced signals are propagated to the nucleus, leading to the activation of the transcription factor CREB, have been characterized. Nerve growth factor (NGF) was found to activate multiple signaling pathways that mediate the phosphorylation of CREB at the critical regulatory site, serine 133 (Ser-133). NGF activates the extracellular signal-regulated kinase (ERK) mitogen-activated protein kinases (MAPKs), which in turn activate the pp90 ribosomal S6 kinase (RSK) family of Ser/Thr kinases, all three members of which were found to catalyze CREB Ser-133 phosphorylation in vitro and in vivo. In addition to the ERK/RSK pathway, we found that NGF activated the p38 MAPK and its downstream effector, MAPK-activated protein kinase 2 (MAPKAP kinase 2), resulting in phosphorylation of CREB at Ser-133. Inhibition of either the ERK/RSK or the p38/MAPKAP kinase 2 pathway only partially blocked NGF-induced CREB Ser-133 phosphorylation, suggesting that either pathway alone is sufficient for coupling the NGF signal to CREB activation. However, inhibition of both the ERK/RSK and the p38/MAPKAP kinase 2 pathways completely abolished NGF-induced CREB Ser-133 phosphorylation. These findings indicate that NGF activates two distinct MAPK pathways, both of which contribute to the phosphorylation of the transcription factor CREB and the activation of immediate-early genes.

Animals↗
PubMed1998

[Determination of diflunisal in plasma by RP-HPLC after solid-liquid extraction].

A RP-HPLC method was developed with solid-liquid extraction technique. Plasma sample was extracted on a macroreticular resin cartridge with methanol--glacial acetic acid (99:1) as elution solvent. After extraction, the assay was carried out on a Spherisorb C18 column with p-phenylphenol as internal standard. The mobile phase is a mixture of methanol--water--glacial acetic acid (66:30:4). UV detection was performed at 250 nm. The flow rate was 1.0 ml.min-1. A good linearity was found at the concentration range from 0.5 to 100 micrograms.ml-1, with the lowest detection limit 0.02 microgram.ml-1 (S/N = 2). The extraction and method recoveries were 91.65% and 97.25% respectively, while the RSD for the within-day and between-day precision were all less than 10%. The above method was applied to determine the plasma concentration of difunisal in three human volunteers after a single oral dosage of 300 mg. Two hours after administration, the plasma concentration of diflunisal reached maximum level. A two compartment method was used to study the pharmacokinetic parameters. The T1/2 alpha and T1/2 beta were 1.40 h and 17.85 h, respectively.

Chromatography, High Pressure Liquid↗
PubMed1998

[Transient response in electric measurement of human auricular point].

This is an experiment on the measuring E approximately t and R approximately t for human auricular point, which shows there is a transient response of electric potential E(t) and voltage drop U(t) when t < 2 tau and there is at intervals change with the time after 2 tau about R(t) and E(t). We have obtained a response function of the E(t) and U(t) by the circuit analysis. The parameter of the transient character is relaxation time tau, tau approximately RC. The mathematical model is in correspondence with the experimental result of the auricular point and TPM simulation. It indicates that the character changing with the time should be extracted after 2 tau. This experiment is of importance to understanding the electric character of the otopoint.

Acupuncture Points↗
PubMed1998

[Transient response in electric measurement of human auricular point].

This is an experiment on the measuring E-t and R-t for human auricular point, which shows there is a transient response of electric potential E(t) and voltage drop U(t) when t < 2 tau and there is at intervals change with the time after 2 tau about R(t) and E(t). We have obtained a response function of the E(t) and U(t) by the circuit analysis. The parameter of the transient character is relaxation time tau,tau approximately RC. The mathematical model is in correspondence with the experimental result of the auricular point and TPM simulation. It indicates that the character changing with the time should be extracted after 2 tau. This experiment is of importance to understanding the electric character of the otopoint.

Acupuncture Points↗
PubMed1997

Conformation of the N-terminal segment of a monocysteine mutant of troponin I from cardiac muscle.

A monocysteine mutant of cardiac muscle troponin I, cTnI(S5C/C81I/C98S), was generated from a mouse cTnI cDNA clone and expressed in a bacterial system. Cys-5 was modified with the fluorescent sulfhydryl reagent IAANS to probe the conformation of the N-terminal extension of the mutant and the mutant complexed with cardiac muscle troponin C. Our emphasis was on the effect of phosphorylation of Ser-23 and Ser-24 by protein kinase A on the conformation of the N-terminal segment. Phosphorylation resulted in an 8-nm red-shift of the emission spectrum of the attached IAANS probe and a reduction of its quantum yield by a factor of 4-5. The intensity decay of nonphosphorylated IAANS-labeled mutant was complex and had to be described by a sum of three exponential terms, with lifetimes in the range 0.1-5 ns. A fourth component in the range 7-9 ns was required to describe the intensity decay of the phosphorylated mutant. Phosphorylation also reduced the weighted mean lifetime, consistent with the changes observed in the steady-state fluorescence parameters and a 33% decrease in the global rotational correlation time calculated from anisotropy decay data. This change in correlation time suggested a decrease in the axial ratio of the protein. The fluorescence changes of the labeled mutant induced by phosphorylation were carried over to its complex with troponin C. The Stern-Volmer plots of acrylamide quenching of the steady-state fluorescence were essentially linear for nonphosphorylated mutant but displayed pronounced concave downward curvatures for the phosphorylated protein under all conditions studied. The present results are interpreted in terms of a more compact hydrodynamic shape of the phosphorylated cTnI mutant and are consistent with a folded conformation of the N-terminal extension induced by phosphorylation of the two serines. These conformational changes may play a role in the modulation of cardiac muscle contractility by troponin I phosphorylation.

Animals↗
PubMed1997

Phosphorylation-induced distance change in a cardiac muscle troponin I mutant.

Phosphorylation of two adjacent serine residues in the unique N-terminal extension of cardiac muscle troponin I (cTnI) is known to decrease the Ca2+-sensitivity of cardiac myofilaments. To probe the structural significance of the N-terminal extension, we have constructed two cTnI mutants each containing a single cysteine: (1) a full-length cTnI mutant (S5C/C81I/C98S) and (2) a truncated cTnI mutant (S9C/C50I/C67S) in which the N-terminal 32 amino acid residues were deleted. We determined the apparent binding constants for the complex formation between IAANS-labeled cardiac troponin C (cTnC) and the two cTnI mutants. The affinities of the cTnC for the truncated cTnI mutant were: (1) 1.5 x 10(6) M(-1) in EGTA, (2) 28.9 x 10(6) M(-1) in Mg2+, and (3) 87.5 x 10(6) M(-1) in Mg2+ + Ca2+. These binding constants were approximately 1.4-fold smaller than the corresponding values obtained with the full-length cTnI mutant, suggesting a very small contribution of the N-terminal extension to the binding of cTnI to cTnC. Cys-5 in the full-length cTnI mutant was labeled with IAANS, and the distribution of the separation between this site and Trp-192 was determined by analysis of the efficiency of fluorescence resonance energy transfer from Trp-192 to IAANS. The following mean distances were obtained with the unphosphorylated full-length mutant: 44.4 A (cTnI alone), 48.3 A (cTnI + cTnC), 46.3 A (cTnI + cTnC in Mg2+), and 51.6 A (cTnI + cTnC in Mg2+ + Ca2+). The corresponding values of the mean distance determined with the phosphorylated full-length cTnI mutant were 35.8, 36.6, 34.8, and 37.3 A. The phosphorylation of cTnI reduced the half-width of the distribution from 9.5 to 3.7 A. Similar but less pronounced decreases of the half-widths were also observed with the phosphorylated cTnI complexed with cTnC in different ionic conditions. Thus, phosphorylation of cTnI resulted in a decrease of 9-12 A in the mean distance between the sites located at the N- and C-terminal portion of cTnI. Our results indicate that phosphorylation elicits a change in the conformation of cTnI which underlies the basis of the phosphorylation-induced modulation of cTnI activity.

Animals↗
PubMed1997

Effects of garlic thioallyl derivatives on growth, glutathione concentration, and polyamine formation of human prostate carcinoma cells in culture.

This study investigated whether naturally occurring garlic derivatives and synthetic S-cysteinyl compounds that resemble garlic constituents have antiproliferative effects on human prostate carcinoma (LNCaP) cells. Studies also examined whether S-allylmercaptocysteine and S-allylcysteine affect two important molecular targets, namely reduced glutathione and polyamines. Results showed that S-allylmercaptocysteine (50 mg/L) diminished LNCaP cell growth whereas the antiproliferative effect of S-allylcysteine was not as pronounced. Studies using synthetic S-cysteinyl analogues revealed that growth inhibition was most effective with compounds containing a disulfide or an active diallyl moiety. Marginal to no inhibitory effect was observed with monosulfinic analogues. Both S-allylmercaptocysteine and S-allylcysteine caused an increase in LNCaP cell reduced glutathione concentrations. Putrescine and spermine concentrations decreased and spermidine increased 3 d after S-allylmercaptocysteine treatment. At 5 d after S-allylmercaptocysteine treatment, polyamine concentrations were similar to those of saline-treated controls. Diminished cell growth and altered polyamine concentrations suggest that S-allylmercaptocysteine may impede the polyamine synthesizing enzyme, ornithine decarboxylase, either by enhancing the formation of reduced glutathione, a known inhibitor of ornithine decarboxylase, or by reacting directly with ornithine decarboxylase at its nucleophilic thiol moiety. Because S-allylcysteine also increases reduced glutathione formation but does not significantly inhibit growth, the latter mechanism may be more likely for this compound. These data provide further evidence that nonessential nutrients derived from garlic may modulate tumor growth. Further research is required on effects of garlic derivatives in vivo before information from the present studies can be used to assist in the development of effective nutritional strategies for preventing progression of prostate cancer.

Antineoplastic Agents, Phytogenic↗
PubMed1997

Multimodal representation of space in the posterior parietal cortex and its use in planning movements.

Recent experiments are reviewed that indicate that sensory signals from many modalities, as well as efference copy signals from motor structures, converge in the posterior parietal cortex in order to code the spatial locations of goals for movement. These signals are combined using a specific gain mechanism that enables the different coordinate frames of the various input signals to be combined into common, distributed spatial representations. These distributed representations can be used to convert the sensory locations of stimuli into the appropriate motor coordinates required for making directed movements. Within these spatial representations of the posterior parietal cortex are neural activities related to higher cognitive functions, including attention. We review recent studies showing that the encoding of intentions to make movements is also among the cognitive functions of this area.

Animals↗
PubMed1997

Tetrandrine inhibited chronic "inflammatory" pulmonary hypertension in rats.

AIM: To study the effects of tetrandrine (Tet), on pulmonary hypertension. METHODS: An "inflammatory" chronic pulmonary hypertension induced by monocrotaline (Mon) in rats was used. RESULTS: Tet 50, 100, and 150 mg.kg-1.d-1 i.g. 3 wk inhibited Mon-induced increase of pulmonary artery pressure (PAP) by 23.8%, 34.9% (P < 0.05), and 42.0%, (P < 0.05); the right heart index by 2.0%, 25.0%, and 30.0% (P < 0.05) respectively compared with those from Mon group, without significant influence on the systemic artery pressure (SAP). Using histological exam by Verhoeff elastic stain and computer scanning analysis, it was found that Tet (100 mg.kg-1.d-1) for 3 wk, inhibited the increase of medial thickness and cross sectional area by 57.8% (P < 0.01) and 54.6% (P < 0.01), respectively vs Mon group. CONCLUSION: Tet ameliorated the development of pulmonary vascular and lung tissue injury induced by Mon in rats.

Alkaloids↗
PubMed1996

Structural studies of kinesin-nucleotide intermediates.

We have investigated the structural changes that occur in the molecular motor kinesin during its ATPase cycle, utilizing two bacterially expressed constructs. The structure of both constructs has been examined as a function of the nature of the nucleotide intermediate occupying the active site by means of sedimentation velocity, sedimentation equilibrium, fluorescence solute quenching, fluorescence anisotropy decay, and limited proteolysis. While the molecular weight of monomeric and dimeric human kinesin constructs, as measured by sedimentation velocity and sedimentation equilibrium, and the tryptic cleavage pattern are unaffected by the nucleotide intermediate occupying the active site, significant changes in the rotational correlation time of fluorescently labeled kinesin-nucleotide intermediates can be detected. These results suggest that kinesin contains an internal "hinge" whose flexibility varies through the course of the ATPase cycle. In prehydrolytic, "strong" binding states, this hinge is relatively rigid, while in posthydrolytic, "weak" binding states, it is more flexible. Our results, in conjunction with anisotropy decay studies of myosin, suggest that these two molecular motors may share a common structural feature; viz. weak binding states are characterized by segmental flexibility, which is lost upon assumption of a strong binding conformation.

Adenosine Diphosphate↗
PubMed1996

Coupling of the RAS-MAPK pathway to gene activation by RSK2, a growth factor-regulated CREB kinase.

A signaling pathway has been elucidated whereby growth factors activate the transcription factor cyclic adenosine monophosphate response element-binding protein (CREB), a critical regulator of immediate early gene transcription. Growth factor-stimulated CREB phosphorylation at serine-133 is mediated by the RAS-mitogen-activated protein kinase (MAPK) pathway. MAPK activates CREB kinase, which in turn phosphorylates and activates CREB. Purification, sequencing, and biochemical characterization of CREB kinase revealed that it is identical to a member of the pp90(RSK) family, RSK2. RSK2 was shown to mediate growth factor induction of CREB serine-133 phosphorylation both in vitro and in vivo. These findings identify a cellular function for RSK2 and define a mechanism whereby growth factor signals mediated by RAS and MAPK are transmitted to the nucleus to activate gene expression.

Amino Acid Sequence↗
PubMed1996

Networks with lateral connectivity. I. dynamic properties mediated by the balance of intrinsic excitation and inhibition.

1. We studied the rapid dynamic changes of neuron response properties in the somatosensory cortex by the use of computer simulations. The model consists of three feedforward layers of spiking neurons, corresponding to skin, subcortex, and cortex structures. Measurements and analysis of model activity throughout this work are similar to those used in neurophysiological experiments. 2. The effects of various parameters on response properties of model neurons were investigated. The most important parameters were the lateral excitation and inhibition in the simulated cortical network. 3. The balance between excitation and inhibition is a key factor in determining the stability of the network model. There is a large excitation-inhibition (E-I) parameter region within which the model can stably respond to inputs. 4. The input-output relations and receptive field (RF) sizes of simulated neurons are modifiable by the E-I balance. The shapes of RFs are determined by both feedforward projections and the spatial distribution of lateral connections. 5. We simulated changes in temporal and spatial properties of neurons in response to manipulations that mimic bicuculine methiodide or glutamate application to the cortex. Simulation results agreed well with experimental data, suggesting that cortical transmitter levels play an important role in the dynamic responses of the neural net through their effects on E-I balance. 6. With parameters of the model set to an inhibition-dominant scheme, the model was able to reproduce experimentally observed rapid RF expansions that follow cortical lesion or input denervation. Simulation results also suggested that spontaneous inputs to a sensory system can serve as a source of tonic inhibition in the cortex. 7. We conclude that lateral connections could produce and maintain a cortical network having dynamic properties without the need to invoke synaptic plasticity. Individual neuron properties could be modified by changing the balance of cortical layer excitation and inhibition. In a real brain, this could be achieved either by changing levels of cortical transmitter (gamma-aminobutyric acid. for example) or by changing tonic background input to the cortical network.

Afferent Pathways↗
PubMed1996

Networks with lateral connectivity. II. Development of neuronal grouping and corresponding receptive field changes.

1. Using a three-layered network model defined in the previous paper, we studied the basic features of neurons in the cortical layer while the synaptic strengths of lateral excitatory connections were made modifiable by a Hebbian learning rule and a normalization process. 2. We found that neurons in the cortical layer formed groups through their lateral excitatory connections after the network was trained with sequential random dot stimulations. Neurons within a group connected tightly; neurons in different groups connected weakly. 3. The effects of model parameters and input parameters on the formation of neuronal groups were investigated. Results showed that the average size and rough shapes of groups were mainly determined by the spatial distribution of lateral connections within the cortical layer, irrespective of input parameters and training methods. Thus groups are structure dependent. 4. Lateral inhibition in the network is the only key factor that affects the grouping of neurons. Without an appropriate amount of distant inhibition, group formation does not occur. Group formation is very robust to all other parameters we tested. On the other hand, group locations are very easily disturbed by inputs or changes of parameters, suggesting that such neuronal groups are dynamically maintained. 5. With the development of neuronal groups, neurons can be divided into two response types. TN-1 neurons respond weakly to inputs and have small receptive fields or do not respond at all (silent); TN-II neurons, approximately 30-40% of all, respond strongly to inputs and have large receptive fields. The two types of neurons also differ with respect to response threshold and temporal firing patterns. After groups formed, receptive fields of TN-II neurons within the same group clustered spatially with high overlap, whereas receptive fields of TN-I neurons with detectable responses shifted systematically with the neuron's spatial location. 6. The two types of neurons are homogeneously distributed across the cortical layer. The population of each type of neuron produces a full representation of the input layer with weak or strong responses, respectively. 7. We concluded that neurons in the cortical network naturally assembled into functional groups. Such groups are dynamic and amenable to change by input stimuli. A fraction of neurons (30-40%) within the same group shares a similar receptive field and strongly respond together to stimuli, so that the network has more robust response to inputs. On the other hand, the responses of a large portion (60-70%) of neurons become weak or silent: these neurons are available for other (unknown) functional purposes.

Afferent Pathways↗
PubMed1996

Networks with lateral connectivity. III. Plasticity and reorganization of somatosensory cortex.

1. Mechanisms underlying cortical reorganizations were studied using a three-layered neural network model with neuronal groups already formed in the cortical layer. 2. Dynamic changes induced in cortex by behavioral training or intracortical microstimulation (ICMS) were simulated. Both manipulations resulted in reassembly of neuronal groups and formation of stimulus-dependent assemblies. Receptive fields of neurons and cortical representation of inputs also changed. Many neurons that had been weakly responsive or silent became active. 3. Several types of learning models were examined in simulating behavioral training, ICMS-induced dynamic changes, deafferentation, or cortical lesion. Each learning model most accurately reproduced features of experimental data from different manipulations, suggesting that more than one plasticity mechanism might be able to induce dynamic changes in cortex. 4. After skin or cortical stimulation ceased, as spontaneous activity continued, the stimulus-dependent assemblies gradually reverted into structure-dependent neuronal groups. However, relationships among individual neurons and identities of many neurons did not return to their original states. Thus a different set of neurons would be recruited by the same training stimulus sequence on its next presentation. 5. We also reproduced several typical long-term reorganizations caused by pathological manipulations such as cortical lesions, input loss, and digit fusion. 6. In summary, with Hebbian plasticity rules on lateral connections, the network model is capable of reproducing most characteristics of experiments on cortical reorganization. We propose that an important mechanism underlying cortical plastic changes is formation of temporary assemblies that are related to receipt of strongly synchronized localized input. Such stimulus-dependent assemblies can be dissolved by spontaneous activity after removal of the stimuli.

Afferent Pathways↗
PubMed1996

[Experimental studies on damage to acoustic organ in guinea pigs caused by imitated coal mining noise].

To study the damage to the hearing of miners under the shaft caused by noise and its recovery, an experiment with imitated noise was carried out in guinea pigs. The animals were divided into five groups with exposure to imitated underground noise of 117 dB (A) for one hour for one, two, four, eight and 16 days, respectively, and one control. Thresholds of cortex reaction and evoked electric reaction of brain stem were detected before and after exposure. And, ultrastructural changes in cochlear hair cells were observed by scanning and transmission electron microscopy. Results showed that exposed noise level and length since removing from exposure had significant effects on changes in hearing threshold. Damage to hearing was aggravated with prolonged exposure, to various extent, from changes in temporary threshold shift and permanent threshold shift to recovery from it after removing from exposure. Intracellular structural changes in internal and external hair cells and their injury deteriorated gradually with increase of exposure levels, at worst, to changes in epithelial plate, collapse of Corti's organ and vacuolation, edema and degeneration of cells. It suggests it is important to protect workers form exposure to noise.

Animals↗