Crepidiastrum denticulatum Extract Ameliorates Kidney Ischemia-Reperfusion Injury in Mice.

BACKGROUND: Crepidiastrum denticulatum (CD) is a well-known, traditionally consumed vegetable in Korea, which was recently reported to contain bioactive compounds with detoxification and antioxidant properties. Ischemia-reperfusion injury (IRI) is a major problem after renal transplantation. Furthermore, inflammatory responses to IRI exacerbate the resultant renal injury. In the present study, we investigated whether CD extract exhibits renoprotective effects against IR-induced acute kidney injury in mice. MATERIALS AND METHODS: Renal IRI was induced in male C57BL/6 mice by bilateral renal pedicle occlusion for 30 minutes followed by reperfusion for 48 hours. CD extract (75 mg/kg) was administered orally 5 days before IRI. RESULTS: Treatment with CD extract significantly decreased blood urea nitrogen and serum creatinine levels as well as kidney tubular injury. CD also prevented IRI-induced renal glutathione depletion and increased malondialdehyde levels. Western blotting and reverse transcriptase polymerase chain reaction indicated that CD extract significantly attenuates inducible nitric oxide synthase and toll-like receptor 2/4 protein levels 48 h after IRI. The expression of tumor necrosis factor-α and interleukin-1ß was significantly decreased in the CD extract treatment group. CONCLUSION: CD extract improved acute renal IRI through its antioxidant and anti-inflammatory effects. These findings suggest that CD extract is a potential therapeutic agent for acute ischemia-induced renal damage.

Protective Effect of Eupatilin Pretreatment Against Hepatic Ischemia-Reperfusion Injury in Mice.

BACKGROUND: Eupatilin, a pharmacologically active flavone derived from Artemisia species, is known to have antioxidant and antiinflammatory activities. Ischemia-reperfusion injury (IRI) is a major critical event that commonly occurs after liver transplantation and resection. Furthermore, inflammatory responses to IRI exacerbate the resultant hepatic injury. In this study, we investigated whether eupatilin protects against IR-induced acute liver injury in mice. MATERIALS AND METHODS: Partial (70%) hepatic IRI was induced in male C57BL/6 mice by portal triad pedicle occlusion for 90 minutes followed by reperfusion for 6 hours. Eupatilin (10 mg/kg body weight, oral) was administered 4 days before the IRI. RESULTS: Treatment with eupatilin significantly decreased serum alanine aminotransferase and serum aspartate aminotransferase as well as liver histologic changes. Eupatilin also prevented hepatic glutathione depletion and increased malondialdehyde levels induced by IRI. Western blotting indicated that eupatilin significantly increased the levels of heat shock protein and B-cell lymphoma 2 protein, attenuated inducible nitric oxide synthase, and cleaved caspase-3 levels 6 hours after IRI. The expression of the Toll-like receptor 2/4, and phosphorylated nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor was significantly decreased in the eupatilin pretreatment group. CONCLUSIONS: Eupatilin improved the acute hepatic IRI by reducing inflammation and apoptosis. These findings suggest that eupatilin is a promising therapeutic agent against acute IR-induced hepatic damage.

5,7-dihydroxy-3,4,6-trimethoxyflavone attenuates ischemic damage and apoptosis in mouse islets.

BACKGROUND: The transplantation of isolated pancreatic islets is a promising treatment for diabetes. 5,7-dihydroxy-3,4,6-trimethoxyflavone (Eupatilin), a pharmacologically active flavone derived from the Artemisia plant species, has been reported to have antioxidant and anti-inflammatory activities. This study examines the hypothesis that preoperative eupatilin treatment can attenuate ischemic damage and apoptosis before islet transplantation. METHODS: Islets isolated from Balb/c mice were randomly divided into 2 groups, and cultured in medium supplemented with or without eupatilin. In vitro islet viability and function were assessed. After treatment with a cytokine cocktail consisting of tumor necrosis factor (TNF)-α, interferon (INF)-γ, and interleukin (IL)-1ß, islet cell viability, function, and apoptotic status were determined. The glutathione (GSH) and nitrous oxide (NO) levels were also measured. Proteins related to apoptosis were analyzed using Western blotting. RESULTS: There was no difference in cell viability between the 2 groups. Islets cultured in the medium supplemented with eupatilin showed 1.4-fold higher glucose-induced insulin secretion than the islets cultured in the medium without eupatilin. After treatment with a cytokine cocktail, glucose-induced insulin release and the total insulin content of the islets were significantly improved in eupatilin-pretreated islets compared with islets not treated with eupatilin. Apoptosis was significantly decreased, and GSH levels were elevated in the eupatilin-pretreated group. Cytokine-only treated islets produced significantly higher levels of NO, iNOS, and caspase-3 than islets pretreated with eupatilin before cytokine treatment. CONCLUSIONS: These results suggest that preoperative eupatilin administration enhances islet function before transplantation and attenuates the cytokine-induced damage associated with NO production and apoptosis.

Protective effect of eupatilin against renal ischemia-reperfusion injury in mice.

BACKGROUND: Eupatilin, a pharmacologically active flavone derived from Artemisia species, is known to have antioxidant and anti-inflammatory activities. Ischemia-reperfusion injury (IRI) is a major complication after renal transplantation, with inflammatory responses to IRI exacerbating the resultant renal injury. In the present study, we investigated whether eupatilin exhibits renoprotective activities against ischemia-reperfusion-induced acute kidney injury in mice. MATERIALS AND METHODS: Renal IRI was induced in male C57BL/6 mice by bilateral renal pedicle occlusion for 30 minutes followed by reperfusion for 48 hours. Eupatilin (10 mg/kg body weight p.o.) was administered 4 days before IRI. RESULTS: Treatment with eupatilin significantly decreased neutrophil gelatinase-associated lipocalin and kidney injury molecule-1 levels in urine, blood urea nitrogen level, and serum creatinine levels, as well as kidney tubular injury. Western blotting indicated that eupatilin significantly increased the levels of heat shock protein 70 and B-cell lymphoma protein, and it attenuated inducible nitric oxide synthase, Bcl-2-associated X protein, and caspase-3 levels 48 hours after IRI. CONCLUSION: Our findings suggest that eupatilin is a promising therapeutic agent against acute ischemia-induced renal damage.

Scaling and organization of electroencephalographic background activity and alpha rhythm in healthy young adults.

The coexistence of the broad-band fluctuation and alpha rhythm of the brain dynamics is studied based on the zero-crossing property of the local electroencephalographic (EEG) recording in eyes closed and eyes open. A two-component zero-crossing scenario, consisting of a broad-band fractal and narrow-band rhythm components, is assumed. Scaling is found in the power law distribution p(tau) approximately tau(-nu) of the crossing time interval tau of the broad-band fluctuation. In alpha dominant brain state, the alpha rhythm interval L also exhibits scaling in the form of power law distribution: p(L) approximately L(phi). Our main result is the relationship nu + phi approximately 3 that characterizes the organization of these two prominent features of the brain dynamics. The possible role of self-organized criticality of punctuated equilibrium in this organization is discussed.

Fractal characteristics of human Parkinsonian neuronal spike trains.

Fractal analysis was applied to human pallidal neuronal spike trains recorded from patients with Parkinson's disease during ablative surgery of the internal segment of the globus pallidus. Fractal dynamics was quantified by computing the scaling exponent with the average wavelet coefficient approach. We observed fractal persistent correlation in the fluctuation of the interspike intervals of neuronal spike trains recorded in the internal segment of the globus pallidus both before and after the administration of dopamine agonist apomorphine. However, there was a significant increase in the scaling exponent during the "on" state after apomorphine administration as compared with the parkinsonian "off" state prior to apomorphine. In addition, we observed a statistically significant decrease in the average firing rate in the transition from the "off" to the "on" state. We conclude that robust fractal dynamics can be observed in single neurons in the human CNS, indicating that human neuronal dynamics of the internal segment of the globus pallidus are essentially a nonlinear and nonequilibrium process, with a long-range correlation or memory extending across many time scales. Accompanying the "on" state after apomorphine administration was an improvement in the long-range persistent correlation as compared with the more random dynamics in the "off" state. A scaling exponent signaling a breakdown or modification in long-range correlation in a single neuron may serve as a useful indicator of a dysfunctional network in the human CNS.

Changes in the complex permittivity during spreading depression in rat cortex.

With recent developments in current density imaging (CDI), it is feasible to utilize this new technique in brain imaging applications. Since CDI's ability to measure changes in current density depends on a concomitant activity-dependent change in the conductivity of the brain tissue, we have examined the changes in complex conductivity during spreading depression (SD) in rodent neocortex using a coaxial probe. SD was chosen because it is often referred to as an animal model of cerebral ischemia and migraine with aura. The conductivity measurements revealed a change with short latency (30-60 s) followed by a change with a longer latency (200-300 s). This change in conductivity with short latency has not been reported before, and we conjecture that it may be the priming or triggering mechanism prior to the main SD episode. A 20% change in conductivity during SD is sufficiently large to be measured by CDI. Therefore, the ability to measure changes in the conductivity, as opposed to metabolic changes, makes CDI a viable approach to the study of ischemia and migraine with aura.

Characterization of cortical spreading depression by imaging of intrinsic optical signals.

The spatiotemporal progression of cortical spreading depression was investigated by imaging of activity-dependent intrinsic optical signals in rat sensorimotor cortex. In contrast to the traditional conception of a uniformly propagating event, the evolution of the spreading depression was characterized by an asymmetrical propagating wavefront and a non-uniform dispersion of optical signals behind the wavefront. As these optical signals are a reflection of underlying molecular and metabolic processes, the imaging approach to spreading depression might constitute a useful technique for investigating the functional organization as well the pathophysiology of the cerebral cortex.

Statistical prediction of the optimal site for thalamotomy in parkinsonian tremor.

Stereotactic lesions in the thalamus for treatment of parkinsonian tremor are often made at the location where neurons fire at approximately tremor frequency (tremor cells). Some of these cells show a large amount of activity at tremor frequency and are significantly correlated with electromyographic activity (EMG) during tremor. Our analysis of cellular location identifies a cluster of neurons showing activity characterized both by concentration of power at tremor frequency and by significant correlation with EMG. In a retrospective analysis of results in 15 patients, lesions placed within 2 mm of the center of this cluster were uniformly effective in relieving tremor. Therefore, a small lesion targeting this cluster is effective in treatment of parkinsonian tremor.

Characteristics of somatotopic organization and spontaneous neuronal activity in the region of the thalamic principal sensory nucleus in patients with spinal cord transection.

1. We explored the region of the principal sensory nucleus of thalamus (Vc) during stereotactic surgical procedures for treatment of patients with pain after spinal cord transection (n = 23). Receptive fields (RFs) of thalamic single neurons and locations of sensations evoked by stimulation (projected field, PF) were determined by standard methods. The cellular thalamic region where sensations were evoked at < 25 microA was termed the "region of Vc." The region of Vc in spinal patients was subdivided into different areas according to RF and PF locations. Areas that were distant from the representation of the anesthetic part of the body were termed "spinal control" areas, whereas those that were adjacent to or included in the representation of the area of absolute sensory loss were termed "border zone/anesthetic" areas. The region of Vc in movement disorder patients were termed the "control" area. 2. Border zone/anesthetic areas of thalamus often exhibited increased representations of the border of the anesthetic part of the body in comparison with the representation of the same parts of the body in control and spinal control areas. 3. In control and spinal control areas the locations of RFs and PFs were usually well matched. However, in border zone/anesthetic areas of the thalamus there was frequently a mismatch between the location of RFs and PFs (RF/PF mismatch). In border zone/anesthetic areas, RFs were often located on the border of the anesthetic part of the body whereas PFs were referred to anesthetic parts of the body. 4. Analysis of first- and higher-order properties of spontaneous neuronal activity revealed that spike trains could be classified into two groups with distinct patterns of activity. The R group (n = 49) was characterized by independence of sequential interspike intervals (ISIs), a Poisson distribution of ISIs, initially inhibitory or flat autocovariance function (acvf), and low level of high-frequency bursting. The O group (n = 26) was characterized by correlation of sequential ISIs, large sustained postspike facilitation on the acvf, and high prevalence of high-frequency bursting--all consistent with a bursting pattern of activity. A third group of spike trains (n = 17) had an initially inhibitory or flat acvf and a unimodal, positively shifted, ISI distribution that did not meet criteria for a Poisson distribution. 5. Spike trains in the R group were much more common in control and control spinal areas, whereas those in the O group were more common in border zone/anesthetic areas.(ABSTRACT TRUNCATED AT 400 WORDS)

Single unit analysis of the human ventral thalamic nuclear group. Tremor-related activity in functionally identified cells.

During procedures for parkinsonian tremor, neurons in the thalamic ventral nuclear group show periodic activity at tremor frequency (tremor-frequency activity). The tremor-frequency activity of some cells is significantly correlated with tremor. Cells in this region also display functional properties defined by activity related to somatosensory stimuli and to active movement. Cells with activity related to somatosensory stimulation were termed sensory cells while those with activity related to active movement were termed voluntary cells. Cells with activity related to both somatosensory stimulation and active movement were termed combined cells. Those with activity related to neither somatosensory stimulation nor active movement were termed no-response cells. Combined, voluntary and no-response cells were located in the region of thalamus where a lesion stops tremor and anterior to the region where sensory cells were found. Spectral cross-correlation analysis demonstrated that many combined, voluntary and no-response cells had a peak of activity at tremor frequency which was significantly correlated with electromyogram (EMG). Analysis of the phase of thalamic activity relative to EMG activity indicated that voluntary and combined cell activity usually led EMG during tremor. These results suggest that thalamic cells unresponsive to somatosensory stimulation (voluntary and no-response cells) and those responsive to somatosensory stimulation (combined cells) are involved in the mechanism of parkinsonian tremor. The activity of sensory cells frequently lagged behind tremor while activity of combined cells often led tremor. This finding suggests that the activity of these two cell types, both responding to sensory input, is related to tremor by different mechanisms.

Neural networks and Parkinson's disease.

A closed-loop or recurrent neural network was taught to generate output discharges to reproduce the prototypical activations in agonist and antagonist muscles which produce the displacement of a limb about a single joint. By introducing a generalized decrease in the excitability of the pre-output layer in the network, the network made the displacement more slowly and also showed an inability to maintain a repetitive movement. These concepts can be applied to the human nervous system in the understanding of the physical basis of movement and its disorders. It is suggested that a movement represents the output of a closed-loop network, such as the cortical-basal ganglia-thalamic-cortical motor loop, which iterates repetitively to its end point or attractor. The model provides an explanation of how the state of thalamic inhibition seen in Parkinson's disease physically may produce bradykinesia and the inability to maintain a repetitive movement.

Mouse mammary tumor virus infection accelerates mammary carcinogenesis in Wnt-1 transgenic mice by insertional activation of int-2/Fgf-3 and hst/Fgf-4.

Transgenic mice carrying the Wnt-1 protooncogene modified for expression in mammary epithelial cells exhibit hyperplastic mammary glands and stochastically develop mammary carcinomas, suggesting that additional events are necessary for tumorigenesis. To induce such events and to identify the genes involved, we have infected Wnt-1 transgenic mice with mouse mammary tumor virus (MMTV), intending to insertionally activate, and thereby molecularly tag, cooperating protooncogenes. Infection of breeding female Wnt-1 transgenics decreased the average age at which tumors appeared from approximately 4 months to approximately 2.5 months and increased the average number of primary tumors per mouse from 1-2 to > 5. A smaller effect was observed in virgin females, and infection of transgenic males showed no significant effect on tumor latency. More than half of the tumors from the infected breeding group contained one or more newly acquired MMTV proviruses in a pattern suggesting that most cells in tumors arose from a single infected cell. Analyses of provirus-containing tumors for induced or altered expression of int-2/Fgf-3, hst/Fgf-4, int-3, and Wnt-3 showed activation of int-2 in 39% of tumors, hst in 3%, and both int-2 and hst in 3%. DNA analyses with probes for protooncogenes and MMTV confirmed that the activations resulted from proviral insertions. There was no evidence for proviral insertions at the int-3, Wnt-3, or Wnt-1 loci. These findings provide further evidence that fibroblast growth factors Int-2 and Hst can cooperate with Wnt-1, another secreted factor, in mammary tumorigenesis, and they illustrate the capacity of this system to identify cooperating oncogenes.

Intramolecular excimer formation of pyrene-labeled lipids in lamellar and inverted hexagonal phases of lipid mixtures containing unsaturated phosphatidylethanolamine.

The rates of intramolecular excimer formation of di(1'-pyrenemyristoyl)phosphatidylcholine (dipyPC) in dioleoylphosphatidylethanolamine (DOPE), egg PE/diolein (DG) and dilinoleoyl-PE (DLPE)/1-palmitoyl-2-oleoyl-PC (POPC) were studied at different temperatures and lipid compositions. Both the excimer-to-monomer intensity ratio and the excimer association rate constant were employed to quantify the rate of excimer formation. The latter was calculated from the measured monomer fluorescence lifetime of dipyPC. We observed that the rate of excimer formation was sensitive to either the temperature-induced or lipid composition-induced lamellar-to-inverted hexagonal phase transition of the above lipid systems. As the lipids entered the inverted hexagonal phase, the rate of excimer formation increased at the temperature-induced phase transition for DOPE, but decreased at the composition-induced phase transition for both TPE/DG and DLPE/POPC systems by increasing the DG% and decreasing the PC%, respectively. We conclude that the rate of intramolecular excimer formation of dipyPC in the non-lamellar phase is influenced both by the intra-lipid free volume of the hydrocarbon region and the intra-rotational dynamics of the two lipid acyl chains.

Automatic end-expiratory air sampling device for breath hydrogen test in infants.

An automatic electronically operated end-expiratory air sampler has been developed for use in small infants. Upon expiration, which is detected by a hot-wire sensor, a small portion of the end-expiratory air is automatically collected into a syringe mounted on a syringe driver. The sampler obtained 87% of the end-expiratory air sample. Additionally, highly reproducible and consistent results were obtained for the respiratory gases (O2, N2, CO2). The sampler has been applied for studying breath hydrogen excretion to detect lactose malabsorption. It may also be applicable to study other expiratory gases in infants.

Single unit analysis of the human ventral thalamic nuclear group. Activity correlated with movement.

During neurosurgical operations for the relief of movement disorders, single thalamic neurons (n = 107) were identified with activity which was related to verbally cued active movements (movement-related cells). The activity of each neuron was examined during different contralateral movements in order to determine the movement which was associated with the most consistent and pronounced change in firing rate (the optimal response). The optimal response was determined by analysis of histograms of neuronal activity which were constructed by using the onset of EMG activity to synchronize successive repetitions of the active movement. Movement-related cells exhibited optimal responses associated with such movements as making a fist, extension or flexion of the wrist, flexing or extending the elbow, pointing with the entire upper extremity, extending the tongue and lifting the leg. Most movement-related cells recorded in a single parasagittal plane in an individual patient had optimal responses related to movements involving the same part of the body. Movement-related cells were classified into those that were activated in response to somatosensory stimulation (combined cells, n = 20) and those which were not (voluntary cells, n = 87). Combined cells were activated in advance of EMG activity during active movement and so could be distinguished from cells responding only to sensory stimulation (sensory cells). Movement-related cells (combined and voluntary cell types) were located anterior to sensory cells and tended to show a mediolateral somatotopic organization parallel to that of sensory cells with cutaneous receptive fields. Combined cells responded to somatosensory stimulation of the same part of the body as that involved in the active movement related to the optimal response of the cell. Combined cells responding to passive movements of a joint always had their optimal response during active movement about the same joint. The activity of combined cells during parkinsonian tremor may clarify the role of sensory feedback in tremor.

Neural network control of simple limb movements.

It is possible to embed the control and computation of a simple single-joint movement at different speeds by a small non-linear network of neuron-like elements. The network "learns" by appropriate adjustment of the strengths of interconnection, or synaptic weights, between the neuron-like elements. The learning of a few movement trajectories is generalized to the learning of a family of unlearned trajectories. These observations are in support of our hypothesis that relaxation of a network from an initial state to a final equilibrium state is both causal and computational to movement generation and control.

Thalamic single-unit activity occurring in patients with hemidystonia.

Lesions of the ventrolateral complex of the human thalamus may relieve abnormal movements in patients with dystonia. We have now recorded the thalamic single-unit activity and the electromyographic (EMG) activity in the upper extremity during the physiologic localization which is required prior to thalamotomy for hemidystonia. The activity of thalamic single units was correlated with the EMG signal by spectral methods. Results of this analysis indicate that a group of thalamic cells show a concentration of activity, occurring at the same frequency as the EMG activity during dystonia. In many cases, there was statistically significant correlation between thalamic and EMG signals at the frequency of dystonia. The activity of these cells may be involved in the generation of dystonic movements.

Characteristics of the bursting pattern of action potentials that occurs in the thalamus of patients with central pain.

Neurons in the somatosensory thalamus of patients with central pain following spinal cord injury fire in bursts of action potentials more frequently than do similar neurons in patients without pain. Furthermore, the characteristic firing pattern within these bursts is similar to that which is shown to be associated with the occurrence of calcium spikes in intracellular studies of thalamic nuclei. This finding may have significant implications for the etiology and treatment of central pain states.