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Biomedical subjects

M Machida

Publications and source records attributed to M Machida.

At least 37 records · Page 2Linked to original sources

Effect of desynchronized inputs on compound sensory and muscle action potentials.

Stimulation of the second (S1) or third (S2) digit elicits a median sensory potential at the wrist. Similarly, a shock applied to the median (Sm) or ulnar (Su) nerve at the wrist evokes a sensory potential of the fourth digit and a muscle potential over the thenar eminence. Hence, a concomitant application of S1 and S2 or Sm and Su with varying interstimulus intervals simulates the effect of desynchronized inputs. In 10 hands, a shift in latency on the order of 1 msec between S1 and S2 or Sm and Su caused a major reduction in sensory potential by as much as 30-40% but little change in muscle action potential. A latency difference slightly less than one-half the total duration of unit discharge maximized the phase cancellation between the two components and consequently the loss of area under the waveform.

Action Potentials

Dissociation of muscle action potentials and spinal somatosensory evoked potentials after ischemic damage of spinal cord.

In patients undergoing spinal fusion and Cotrel-Dubousset instrumentation we recorded compound muscle action potentials (CMAP) from the lower limb and spinal somatosensory evoked potentials (SSEP) from the caudal epidural space after direct stimulation of rostral spinal cord via epidural electrodes. In three of 30 patients tested, the derotation maneuver altered CMAP but not SSEP. In ten dogs, we observed similar dissociation with decrease or disappearance of CMAP amplitude and unchanged SSEP after ligation of the thoracoabdominal aorta or intercostal arteries at each level. In contrast, both CMAP and SSEP were unchanged by clamping the artery at the lumbar level. This is likely due to the lack of collateral vascular flow at the thoracic cord level, the anterior cord in particular, which is mainly supplied by a single large radicular artery (Adamkiewicz artery). These findings support that the CMAP and SSEP are mediated through two independent pathways located in the anterior and posterior spinal cord, respectively. We postulate that the dissociate alteration of CMAP and SSEP by derotation maneuver is due to greater vulnerability of the anterior cord or motor tract to ischemia caused by the displacement of anterior spinal or radiculomedullary artery. Therefore, the patients requiring major derotation procedure would benefit from CMAP monitoring, which provides more sensitive measure of anterior cord function that the conventional SSEP monitoring.

Action Potentials

Nucleotide sequences of Saccharomycopsis fibuligera genes for extracellular beta-glucosidases as expressed in Saccharomyces cerevisiae.

We isolated two genes for extracellular beta-glucosidase, BGL1 and BGL2, from the genomic library of the yeast Saccharomycopsis fibuligera. Gene products (BGLI and BGLII) were purified from the culture fluids of Saccharomyces cerevisiae transformed with BGL1 and BGL2, respectively. Molecular weights of BGLI and BGLII were estimated to be 220,000 and 200,000 by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The two beta-glucosidases showed the same enzymatic characteristics, such as thermo-denaturation kinetics and dependencies on pH and temperature, but quite different substrate specificities: BGLI hydrolyzed cellobiose efficiently, but BGLII did not. This result is consistent with the observation that the S. cerevisiae transformant carrying BGL1 fermented cellobiose to ethanol but the transformant carrying BGL2 did not. Southern blot analysis revealed that the two beta-glucosidase genes were derived from Saccharomycopsis fibuligera and that the nucleotide sequences of the two genes are closely related. The complete nucleotide sequences of the two genes were determined. BGL1 and BGL2 encode 876- and 880-amino-acid proteins which were shown to be highly similar to each other. The putative precursors begin with hydrophobic segments that presumably act as signal sequences for secretion. Amino acid analysis of the purified proteins confirmed that BGL1 and BGL2 encode BGLI and BGLII, respectively.

Amino Acid Sequence

[Urinary beta 2-microglobulin as an indicator for impaired excretion of methotrexate].

Twenty cases treated with high-dose methotrexate (MTX) infusions were studied to determine whether urinary beta 2-Microglobulin (beta 2-M) is a reliable indicator to predict impaired excretion of methotrexate. Before and after MTX infusions, the levels of urinary beta 2-M were measured along with serum creatinine, uric nitrogen, and creatinine clearance. MTX clearance was found to be impaired in four of the 20 infusions, although concentrations of serum creatinine and uric nitrogen, and creatinine clearance were normal prior to the infusions. In the four cases that resulted in impaired excretion, a significant increase of beta 2-M levels was noted before and after the high-dose MTX infusion (p less than 0.01). Although one of the four cases showed a normal level of beta 2-M before the infusion, the post-infusion level of beta 2-M extremely high. This raised level of beta 2-M was thought to be caused by impaired excretion due to decreased amounts of urine. We conclude that urinary beta 2-M is a sensitive indicator of impaired excretion of high-dose MTX infusion, and especially when levels of more than 250 ng/ml of urinary beta 2-M are found to exist, it seems to difficult to clear the drug following high-dose MTX treatment.

Blood Urea Nitrogen

Monitoring of motor action potentials after stimulation of the spinal cord.

We recorded motor action potentials in cats, using surface electrodes placed over the soleus muscle. The action potentials were generated by stimulating the spinal cord with electrodes in the epidural space at the level of the fifth or sixth thoracic vertebra. This also was done in humans, using the same methods of stimulating and recording, but the intensity of the stimulus was adjusted to produce little or no twitch of the paraspinal muscles. In the animal experiment, the motor action potential was abolished after transection of the pyramidal tract and was progressively attenuated with effective doses of a curare-like agent. We also tested the effect of distraction, using the same technique as is used in Harrington instrumentation, and found that the amount of distraction that caused reduction of the amplitude of the motor action potential of more than 50 per cent, when sustained for longer than seven minutes, caused permanent paraplegia in two cats. The evaluation of spinal evoked potentials that were obtained from epidural electrodes placed caudad to the level of distraction, and of motor action potentials that were recorded over the soleus muscle, following the same stimulus, showed a similar pattern of reduction after distraction in five of seven cats. The other two cats had irreversible reduction of motor action potential associated with unchanged spinal evoked potential, and both cats became paraplegic.(ABSTRACT TRUNCATED AT 250 WORDS)

Action Potentials

A positive regulatory sequence of the Saccharomyces cerevisiae ENO1 gene.

ENO1-'lacZ fusions with various lengths of the ENO1 5'-flanking region were constructed on various types of yeast plasmid vectors. The fully expressed level of beta Gal directed by ENO1-'lacZ fusions differed depending on the type of vector, but on any type of vector, beta Gal activity was not greatly influenced by the carbon source in the medium. The 86-bp DNA region of ENO1 at position -487 to -402 upstream of the initiation codon, in which we had previously delimited the positive regulatory region of ENO1 (Uemura, H., Shiba, T., Paterson, M., Jigami, Y., & Tanaka, H. (1986) Gene 45, 67-75), exerted its function without requiring precise location with respect to the TATA box. The action of the positive regulatory region was not affected by its orientation. In addition, the substitution of the UASs of PHO5, encoding repressible acid phosphatase, with the regulatory region of ENO1 changed the expression of PHO5-'lacZ gene to constitutive, irrespective of the concentration of inorganic phosphate in the medium. Furthermore, the GCR1 gene cloned in a multicopy plasmid increased the expression of the ENO1-'lacZ fused genes.

Escherichia coli

[Therapy of renal cell carcinoma. 3. Interferon therapy].

Human lymphoblastoid interferon (HLBI-alpha) or recombinant human leukocyte interferon (HLIF-alpha) was administered to ten patients suffering from renal cell carcinoma at a daily dose of 5 X 10(6) units. The efficacy of interferon was assessed in nine patients who had received HLBI-alpha or HLIF-alpha for more than eight weeks. One patient (11%) demonstrated complete response (CR), three patients (33%) showed no change and five patients (56%) continued to have progressive disease. CR was obtained after 30 weeks under HLBI-alpha and FT-207 combination therapy. The main side effects were fever, anorexia, general fatigue and hematologic toxicities. Cessation of interferon therapy due to side effects was necessary in three patients. In conclusion interferon is one of the most valuable agents in the treatment of renal cell carcinoma.

Adult

Nucleotide sequence and characteristics of the gene for L-lactate dehydrogenase of Thermus caldophilus GK24 and the deduced amino-acid sequence of the enzyme.

The gene for L-lactate dehydrogenase (LDH) (EC 1.1.1.27) of Thermus caldophilus GK24 was cloned in Escherichia coli using synthetic oligonucleotides as hybridization probes. The nucleotide sequence of the cloned DNA was determined. The primary structure of the LDH was deduced from the nucleotide sequence. The deduced amino acid sequence agreed with the NH2-terminal and COOH-terminal sequences previously reported and the determined amino acid sequences of the peptides obtained from trypsin-digested T. caldophilus LDH. The LDH comprised 310 amino acid residues and its molecular mass was determined to be 32,808. On alignment of the whole amino acid sequences, the T. caldophilus LDH showed about 40% identity with the Bacillus stearothermophilus, Lactobacillus casei and dogfish muscle LDHs. The T. caldophilus LDH gene was expressed with the E. coli lac promoter in E. coli, which resulted in the production of the thermophilic LDH. The gene for the T. caldophilus LDH showed more than 40% identity with those for the human and mouse muscle LDHs on alignment of the whole nucleotide sequences. The G + C content of the coding region for the T. caldophilus LDH was 74.1%, which was higher than that of the chromosomal DNA (67.2%). The G + C contents in the first, second and third positions of the codons used were 77.7%, 48.1% and 95.5% respectively. The high G + C content in the third base caused extremely non-random codon usage in the LDH gene. About half (48.7%) the codons in the LDH gene started with G, and hence there were relatively high contents of Val, Ala, Glu and Gly in the LDH. The contents of Pro, Arg, Ala and Gly, which have high G + C contents in their codons, were also high. Rare codons with U or A as the third base were sometimes used to avoid the TCGA sequence, the recognition site for the restriction endonuclease, TaqI. Two TCGA sequences were found only in the sequence of CTCGAG (XhoI site) in the sequenced region of the T. caldophilus DNA. There were three segments with similar sequences in the two 5' non-coding regions, probably the promoter and ribosome-binding regions, of the genes for the T. caldophilus LDH and the Thermus thermophilus 3-isopropylmalate dehydrogenase.

Amino Acid Sequence

What determines the latency and amplitude of stationary peaks in far-field recordings?

In 20 radial nerves from 10 healthy persons, a referential derivation from the tip of the first or second digit registered two biphasic stationary peaks, PI-NI and PII-NII, following stimulation of the nerve in the forearm. These two peaks occurred slightly before the arrival of the propagating impulse at the wrist and at the base of the digit, respectively. With stepwise reduction of stimulation from a maximal to a threshold intensity, the far-field potential decreased in amplitude linearly with the near-field potential recorded at the junction of the volume conductor. Abduction of the first digit or flexion of the second and third digits altered the waveform and, to a lesser degree, the latency of the far-field peaks. However, these changes appeared in an inconsistent manner, which we were unable to characterize. We conclude that in far-field recording a stationary peak results at the border of the adjoining volume conductors in proportion to the magnitude of the propagating axonal volley approaching the boundary.

Adult

New approach for diagnosis in herniated lumbosacral disc. Dermatomal somatosensory evoked potentials (DSSEPs).

Dermatomally activated somatosensory evoked potentials (DSSEPs) were recorded over the scalp of 50 healthy subjects and 40 patients who later underwent exploration for herniated lumbosacral disc. DSSEPs normally consisted of three negative peaks, N40, N56, and N78, and N41, N59, and N82 after bilateral stimulation of L5 and S1 dermatomes, respectively. In the patient group, the first two peaks were often abnormal in the presence of a clinical sensory deficit, whereas the third peak tended to remain normal. At surgery, herniated disc was confirmed in all but six patients. The findings in the myelogram were misleading in eight patients, showing either false-positive or false-negative results. The DSSEPs were normal in six patients with documented disc protrusion at the level of the L5 or S1 root. The test was also normal in another case with herniation at the level of the S2 root. In this series, DSSEPs accurately predicted the level and degree of lumbosacral root involvement. In addition to myelography and other electrodiagnostic tests, the DSSEPs are valuable non-invasive diagnostic tool for the evaluation of herniated lumbosacral disc.

Adolescent

Pharmacokinetics of a new quinolone, AM-833, in mice, rats, rabbits, dogs, and monkeys.

The pharmacokinetics of AM-833 [6,8-difluoro-1-(2-fluoroethyl)-1, 4-dihydro-7-(4-methyl-1-piperazinyl)-4-oxo-3-quinolinecarboxylic acid] were studied in mice, rats, rabbits, dogs, and monkeys by reversed-phase high-performance liquid chromatography. AM-833 was rapidly and completely absorbed from the digestive tracts of mice, rats, and dogs. About half of AM-833 bound to rat and dog serum proteins. Drug levels in lung, spleen, liver, and kidney tissues of rats and dogs were greater than the respective levels in serum but lower in brain tissue. Drug levels in tissues declined with the decrease in levels in serum. AM-833 penetrated rapidly and well into inflammatory exudate of rats. Elimination half-lives in serum were species dependent, ranging from 1.57 h in rabbits to 9.42 h in dogs. Profiles of drug levels in serum were dose related over a single dose range from 2 to 40 mg/kg and not modified significantly during multiple dosing in dogs. Unchanged AM-833 was excreted in urine and bile in both rats and dogs. The metabolism of AM-833 was suggested by evidence that 24-h total recovery of unchanged AM-833 in urine and bile accounted for about half of the intravenous dose in rats.

Animals

Relation between size of compound sensory or muscle action potentials, and length of nerve segment.

In 24 median nerves from 12 healthy subjects, antidromic digital sensory potentials progressively diminished in size, averaging 40.4, 37.0, 30.7, and 23.9 microV X msec with stimulation at the palm, wrist, elbow, and axilla, respectively. In contrast, compound muscle action potentials changed minimally, measuring 19.4, 19.8, 19.0, and 18.2 mV X msec, respectively. Similar studies of the ulnar and radial nerves showed identical trends. Physiologic temporal dispersion can mimic conduction block of sensory nerves by summating the peaks of opposite polarity generated by fast- and slow-conducting axons. This type of cancellation affects muscle responses much less because motor unit potentials of longer duration superimpose nearly in phase, given the same latency shift as the sensory potentials.

Action Potentials

[Effects of cisapride on lower esophageal sphincter pressure and gastroduodenal motor activity in man].

Manometric study was performed to investigate the effects of cisapride, a new non-antidopaminergic gastrointestinal prokinetic compound, on interdigestive lower esophageal sphincter pressure (LESP) and gastroduodenal motility using infused catheter technique. The subjects consisted of 9 healthy volunteers and 29 patients with progressive systemic sclerosis (19), reflux esophagitis (8) and others (2). 4 mg of cisapride was given by bolus injection, continuous infusion or oral administration. The following results were obtained: Intravenous and oral cisapride increased LESP compared with basal pressure. Especially, bolus injection of cisapride caused a significant elevation of LESP during 30 minutes after administration. After administration of cisapride, gastroduodenal motility was accelerated gradually, then inducing IMC-like contractions. By bolus injection, IMC-like contractions were induced in healthy subjects more frequently than in patients group. On the other hand, motility index of stomach and duodenum showed persistent increase in patients group compared with healthy subjects. Cisapride-induced IMC-like contractions initiated from LES and upper part of stomach and mediated to duodenum, though aborad migration was not confirmed in the present study.

Adolescent

Effects of dietary methionine, cystine, and glycine on endogenous hypercholesterolemia in hepatoma-bearing rats.

The effects on hypercholesterolemia of dietary additions of cystine (Cys), methionine (Met), glycine (Gly), and a combination of Met and Gly to a 20% casein diet were studied in male Donryu rats subcutaneously implanted with an ascites hepatoma line of AH109A cells. The hepatoma-bearing rats fed the 20% casein diet lapsed into both endogenous hypertriglyceridemia and hypercholesterolemia when compared to hepatoma-free (normal) rats fed the same diet. The hypercholesterolemia was due to an elevation (3.2 fold) in the very low-density lipoprotein plus low-density lipoprotein (VLDL + LDL)-cholesterol (Ch) level. The high-density lipoprotein (HDL)-Ch level was slightly but significantly decreased. These lipoprotein changes in hepatoma-bearing rats resulted in a marked (4.5 fold) increase in the atherogenic index (AI, (VLDL + LDL)-Ch/HDL-Ch) in comparison with that of tumor-free rats. The dietary additions of 1.2% Met, 1.2% Cys, and a combination of 1.2% Met and 2.5% Gly significantly suppressed the hepatoma-induced increase in (VLDL + LDL)-Ch with no influence on the hepatoma-induced decrease in HDL-Ch, leading to a noticeable fall in AI. These results indicate that hepatoma-bearing rats are useful as an endogenously hyperlipidemic model and that some dietary amino acids are capable of improving hepatoma-induced hypercholesterolemia and abnormal serum lipoprotein profiles.

Animals