Downregulation of nitric oxide in the brain of mice during their hypoxic preconditioning.

An animal model of hypoxic preconditioning was produced in mice by repeated exposure to autohypoxic condition. The animals' tolerance times to hypoxia were 1.7, 1.8, 2.1, and 2.3 times longer in runs 2, 3, 4, and 5, respectively, than that in run 1, and their oxygen consumption and heart and respiration rates were progressively and significantly slowed down during the repetitive exposure to hypoxia. L-Arginine concentration, nitric oxide (NO) synthase-positive cells, NO synthase activity, and NO content in the whole brain and the subregions telencephalon, diencephalon, and brain stem were significantly increased during the first exposure and were, instead of continuing to increase, significantly decreased in run 4 after the second and third exposure. Tolerance times under the hypoxic condition were significantly shortened and prolonged when preadministration of L-arginine and its analog, respectively, was made. These results indicate that NO in the brain is downregulated under condition of hypoxic preconditioning and negatively involved in increased tolerance to hypoxia.

A discrete amino terminal domain of Kv1.5 and Kv1.4 potassium channels interacts with the spectrin repeats of alpha-actinin-2.

The interaction between the amino terminus of Kv1-type potassium channels and alpha-actinin-2 has been investigated. Using a combination of yeast two-hybrid analysis and in vitro binding assays, alpha-actinin-2 was found to bind to the N-termini of both Kv1.4 and Kv1.5 but not to the equivalent segments of Kv1.1, Kv1.2 or Kv1.3. Deletion analysis in the in vitro binding assays delineated the actinin binding region of Kv1.5 to between amino acids 73 and 148 of the channel. The Kv1.5 binding sites in alpha-actinin-2 were found to lie within actinin's internal spectrin repeats. Unlike the reported interaction between actinin and the NMDA receptor, calmodulin was found to have no effect on actinin binding to Kv1.5.

Changes in the contents of glycogen and lactate in the brain and blood during hypoxic preconditioning.

Mice were randomly divided into groups H4 (hypoxic preconditioning group with repetitive hypoxic exposures for four runs), H(1) (hypoxic control group with exposure to hypoxia for one run) and H(0) (normal control group with no exposure to hypoxia). Glycogen content of whole brain of group H4 was found to be significantly higher than that of groups H(1) and H(0). The glycogen content in telencephalon, diencephalon and pons of group H4 was markedly higher than that in the corresponding areas of groups H(1) and H(0). Glycogen content of whole brain in group H(1) was markedly lower than that in group H(0), whereas no significant difference was seen in these brain subregions between groups H(1) and H(0). Brain lactate contents of groups H4 and H(1) did not show significant difference, though they were significantly higher than that of group H(0). Blood lactate content of group H4 was significantly lower than those of groups H(1) and H(0). The results above indicate that the concomitant increase of glycogen and decrease of lactate in the brain are due to the participation of aerobic metabolism during hypoxic preconditioning or the formation of tolerance to hypoxia.

Changes in glycine content in mouse brain during hypoxic preconditioning.

Glycine content of the whole brain and subregions was measured by high pressure liquid chromatography (HPLC) in mice during their repetitive exposures to hypoxia. The contents of glycine in whole brain, diencephalon, and hippocampus and brain stem was significantly increased as the animals tolerance to hypoxia was increased. The results suggest that glycine, as an inhibitory neurotransmitter in the brain, may positively contribute to the development of hypoxic preconditioning.

[Realization of voice coil motor control with H8/338 micro-controller unit in left ventricular assist pump].

In this paper, we present a PID algorithm with velocity project of voice coil motor controlled on the base of H8/338MCU, and we find a driving part, which it is smaller, lighter and more reliable, for the left ventricular assist device. By means of the simulated experiment on the system, the result is satisfying. So we can say that it is a good help for the realization of LVAD.

[Effect of sodium nitroprusside on the spontaneous and induced responses of rat spinal cord dorsal horn neurons].

The present study was performed on anesthetized and paralyzed Wistar rats. Extracellular recordings were made from the lumbar enlargement of the spinal cord using carbon filament electrodes. NO donor sodium nitroprusside (SNP) was locally microdialyzed to the spinal cord, and effects of NO on spontaneous response and the responses induced by mechanical stimulation of the hind-foot were observed. After dialyzing for 10~20 min, SNP at a concentration of 1 micromol/L increased the induced responses to innocuous mechanical stimulation and decreased those to noxoius mechanical stimulation. The induced responses to both innocuous and noxious mechanical stimulations were all decreased after dialyzing the same dose of SNP for 20~30 min. The decrease was shown within 7~15 min dialysis when SNP was used at a concentration of 20 micromol/L. SNP at both concentrations of 1 and 20 micromol/L increased spontaneous spikes of the recorded spinal units. These effects were shown preferentially in deep units. The data suggest that in the deeper neurons of the spinal cord NO of different concentrations has different effects on transmission of nociceptive and nonnociceptive signals induced by mechanical stimulation, and No is involved in the antinociception in spinal neurons and facilitates their spontaneous activity.

[Interaction between pelvic and pudendal afferent inputs in lumbar-sacral spinal cord in rats].

Time-dependent inhibition is one of the means to study the interactions between peripheral inputs. The present study was performed on anesthetized and paralyzed Wistar rats using the technique of conditioning-testing stimulation. Electric stimulation (1.5 3 folds of the threshold intensity) was given to pelvic and pudendal nerves. Extracellular recordings were made from convergent neurons at L(6)-S(1) segments of the spinal dorsal horn. Stimulus intervals between conditioning and testing stimulation were measured when a half of the testing responses were inhibited by conditioning responses or the inhibition just began to occur. The time-dependent inhibition was seen in neurons situated deeper than 300 m beneath the dorsal surface of the spinal cord, and not in more superficial neurons. The inhibition intervals were in the range of 1 360 ms and became longer when conditioning stimulation was given to the pelvic nerve. The inhibition intervals were 1 3 ms in superficial neurons (<300 micrometer) and no apparent time-dependent inhibition occurred. The inhibition in deeper neurons was partially reduced by cold block conducted at segments C(5-6) and the blockage was more significant when conditioning stimulation was applied to the pelvic nerve. These findings suggest that the inputs from the pelvic and pudendal nerves may converge on single neurons at deeper lumbosacral dorsal horn and the pudendal nerve induced responses are more likely to be inhibited in these neurons, which may be further strengthened by superspinal structures.

Effects of ketamine on neuronal activity of the spinal dorsal horn in rats with unilateral hindpaw inflammation.

A total of 32 units were extracellularly recorded from the spinal dorsal horn of rats. Unitary discharges evoked by stimulation of A and C fiber in ipsilateral lateral and medial plantar nerve were increased after carrageenan injection to the plantar area. The evoked responses to both A and C fiber were significantly decreased or even disappeared after administration of ketamine. The windup phenomenon was observed in neurons located deeply in the dorsal horn following carrageenan injection and was significantly suppressed or abolished after ketamine administration. The results above show NMDA receptor appears to be involved in the increase of excitability and the development of windup phenomenon in the spinal cord dorsal horn associated with carrageenan induced inflammation.

Changes of adenosine and its A(1) receptor in hypoxic preconditioning.

Effects of hypoxic preconditioning on adenosine (ADO) and its A(1) receptor were studied in Kunming mice. The ADO content and its metabolites in the brain were measured by a specific enzymatic method; a radioligand binding method was used to study the ADO A(1) receptor. The ADO content of the hippocampus in group C (exposure to 4 runs of hypoxia) was markedly higher than that in group A (control, without exposure to hypoxia and B (exposure to 1 run of hypoxia), showing that the ADO content could be cumulatively increased in the hippocampus, which was more sensitive to ischemia and hypoxia, during acute and repeated exposure to hypoxia. A(1) receptor density in group C was significantly lower than in group A and no difference was seen between groups B and C; A(1) receptor affinity in the hippocampus, pons and medula oblongata in group C was significantly higher than in group A, implying that during hypoxic preconditioning there might be some mechanisms preventing A(1) receptor density from decreasing further and making A(1) receptor affinity increase in some brain regions. These results indicate that cumulatively increased ADO in the hippocampus via A(1) receptor may play a neuroprotective role in the CNS as an inhibitory neuromodulator and thus contribute to the formation and development of acute hypoxic adaptation or tolerance.

Changes of superoxide dismutase, glutathione perioxidase and lipid peroxides in the brain of mice preconditioned by hypoxia.

We have developed an animal model of hypoxic preconditioning and assumed that oxygen radicals and their endogenous scavenging enzymes may play an important role in the preconditioning. To test this hypothesis, activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and the content of lipid peroxides (LPO) were measured during the preconditioning. Compared with unpreconditioned control animals, in animals exposed to hypoxia only once, the activities of SOD and GSH-Px in whole brain were found to be significantly decreased while the LPO content in the hippocampus significantly increased. However, those in animals exposed to 4 runs of hypoxia tended to return to control levels and were lower than those in animals exposed to 1 and 2 runs of hypoxia. Our results indicate that the oxygen radicals and their specific scavenging enzymes seem to be involved in the development of tolerance to hypoxia.

Alteration of oxygen consumption and energy metabolism during repetitive exposure of mice to hypoxia.

Changes in oxygen consumption, body temperature and energy metabolism were studied while mice were repeatedly exposed to a sealed environment. The average tolerance limits of environmental oxygen level (vol%) and the average oxygen consumption rates (ml/g x min) were exponentially decreased and the average body rectal temperatures (degrees C) were linearly declined while the average tolerable times (min) to hypoxia were linearly increased as animals were repeatedly exposed to hypoxia for 5 runs. The average survival times (min) in sealed environments after administration of normal saline, iodoacetic acid, malonic acid, potassium cyanide, and potassium cyanide plus iodoacetic acid in group exposed repeatedly to hypoxia for three runs were, respectively, 3.1, 3.9, 1.4, 2.6, and 2.8 times those of the control groups that had corresponding administration of the different chemicals, but no exposure to hypoxia. The results indicate that progressive increase in hypoxia tolerance is related to progressively lower rate of oxygen consumption and heat production, and the lowered energy requirement during repetitive exposure to hypoxia is achieved mainly via pathways of the respiratory chain and glycolysis.

Branching and/or collateral projections of spinal dorsal horn neurons.

Branching and/or collateral projections of spinal dorsal horn neurons is a common phenomenon. Evidence is presented for the existence of STTm/STTl, STTc/STTi, STT/SMT, STT/SRT, SCT/DCPS, SST/DCPS, SCT/SST, STT/SHT, STeT/SHT, STeTs and other doubly or multiply projecting spinal neurons that have been anatomically and physiologically identified and named based on the locations of the cells of origin and their terminations in the brain. These newly discovered spinal projection neurons are characterized by a single cell body and branched axons and/or collaterals that project to two or more target areas in the brain. These novel populations of neurons seem to be a fuzzy set of spinal projection neurons that function as an intersection set of the corresponding single projection spinal neurons and to be at an intermediate stage phylogenetically. Identification strategies are discussed, and general concluding remarks are made in this review.

Pharmacokinetic and local tissue disposition studies of naproxen-following topical and systemic administration in dogs and rats.

The pharmacokinetic profiles of naproxen in blood and synovial fluid (SF) following topical and i.v. bolus administration in dogs, and the local tissue disposition of the drug following topical and oral administration in rats, were investigated to assess the feasibility of topical delivery of naproxen for local and systemic effects. The naproxen gel in poloxamer 407 (PF-127) was applied on the stifle joint of dogs, and serum and synovial fluid samples were collected. For local tissue disposition studies, the naproxen gel was applied on the dorsal skin in rats, and blood, skin, and muscle samples were taken at 3, 6, and 12 h postdose after removing the residual gel from the skin. Steady state serum concentrations occurred at approximately 20 h after topical doses and lasted for the next approximately 30 h in dogs. Similar SF-serum concentration ratios of naproxen were found between i.v. (0.61 +/- 0.16) and topical (0.55 +/- 0.14) routes of administration. Following the i.v. dose, the half-life of naproxen in SF (approximately 60 h) was significantly longer than that in serum (approximately 40 h). The bioavailability of naproxen in the topical gel was approximately 2% of the applied dose in dogs. A large accumulation of drug in the epidermis, dermis, and muscle tissue beneath the gel application site was found in rats. Isopropyl myristate (IPM) significantly increased the systemic absorption as well as the concentrations of naproxen in the underlying dermis and muscle tissues, but exerted little effect on the disposition of naproxen in the epidermis.

[Molecular mechanisms underlying gating activity of voltage dependent ion channels].

A model of two-channel and its activation and inactivation proposed by Hodgkin and Huxley in 1950s have been continuously confirmed by molecular biological and electrophysiological studies since 1980s. The activation of Na+ and K+ channels seems to be dependent on screw-like rotation outward of S4 segment which is highly conservative and very rich in positively charged amino acid residues. The inactivation of Na+ channels appears to be related to "hinged lids" like movement of intracellular linker between domain III and IV; type N-, C-, and P- of inactivation can be divided in K+ channels, which seems to occur mainly in terminal N-, C- and region P, respectively and the N-type inactivation seems to be related to "ball and chain" like movement of terminal N.