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Biomedical subjects

J Fukushima

Publications and source records attributed to J Fukushima.

At least 91 records · Page 5Linked to original sources

Elastase gene expression in non-elastase-producing Pseudomonas aeruginosa strains using novel shuttle vector systems.

In order to determine whether non-elastase-producing strains of Pseudomonas aeruginosa such as N-10, PA103 and IFO3080 can express foreign elastase genes, we introduced elastase genes from P. aeruginosa IFO3455 (elastase-producing) as well as from PA103 and N-10 into non-elastase-producing P. aeruginosa strains. Results suggested that gene expression, secretion, and precursor processing systems of elastase were essentially normal in P. aeruginosa N-10 and IFO3080. Our studies using various elastase genes showed that both the elastase structural gene and 5'-upstream regions of P. aeruginosa PA103 were also normal. This was confirmed by the finding that P. aeruginosa N-10 and IFO3080 which carry the PA103 elastase gene produced elastase. Several deleted or chimeric genes were constructed using the 5'-upstream regions of elastase genes from P. aeruginosa N-10 or PA103 and studies of expression revealed that two individual DNA bases seem to be important in suppressing P. aeruginosa N-10 elastase gene expression. Possible reasons for the lack of elastase expression in these non-elastase-producing strains are discussed.

Base Sequence↗

Effects of chemical deactivation of the interstitial nucleus of Cajal on the vertical vestibulo-collic reflex induced by pitch rotation in alert cats.

The basic circuitry for the vestibulo-collic reflex (VCR) is a three-neuron arc, and this reflex requires the temporal and spatial transformation of vestibular signals to activate the appropriate neck muscles. Signals carried by vestibulo-collic neurons are insufficient to explain the responses of neck muscles. However, it is still unknown as to where the additional signal conversion is performed in the vertical VCR. We examined the effects of chemical deactivation of the interstitial nucleus of Cajal (INC) on the responses of biventer cervicis EMG induced by pitch rotation in the dark in alert head-fixed cats, and compared the results with the vertical vestibulo-ocular reflex (VOR) and also with the VCR and VOR induced by horizontal rotation. Muscimol infusion into the bilateral INC resulted in phase advance and gain drop in both the vertical VCR and the VOR, although the change was smaller in the VCR. The response phases of the horizontal VCR and VOR were not affected. Muscimol infusion outside the INC did not affect the phase of the vertical VCR or VOR. These results suggest that the INC is partially involved in temporal conversion of vestibular signals in the vertical VCR as well as in the VOR evoked by pitch rotation in alert cats.

Animals↗

Relationship between antisaccades and the clinical symptoms in Parkinson's disease.

We studied voluntary control of saccadic eye movement in 32 parkinsonian patients using the antisaccade task (the subjects were instructed not to look at the target but to look in the opposite direction). Mean latencies and error rates in the antisaccade task were significantly increased in advanced parkinsonian patients. The latencies of the antisaccades correlated with the severity of bradykinesia as well as the results of the Wisconsin Card Sorting Test. Patients taking anticholinergics showed significantly higher error rates. These results suggest that disturbance in the initiation of voluntary saccades in advanced patients may be associated with frontal lobe dysfunction, while anticholinergics may affect the inhibitory control of reflexive saccades.

Analysis of Variance↗

Portal hypertension in schistosomiasis: a long-term follow-up of a randomized trial comparing three types of surgery.

The long-term follow-up of patients with the severe form of Manson's schistosomiasis who had had elective surgical treatment for portal hypertension, in a randomized trial, was clinically evaluated. Of 94 patients, proximal splenorenal shunting was performed in 32, esophagogastric devascularization with splenectomy in 32 and distal splenorenal shunting in 30. Patients were observed during a mean of 85.7 +/- 33.1 mo, excluding nine patients (9.6%) who were lost to follow-up. Recurrence of upper gastrointestinal tract bleeding occurred in 24.1% of the patients, without statistical differences among the three groups, but rebleeding because of varices was more frequent after esophagogastric devascularization with splenectomy. Hepatic encephalopathy was significantly higher after proximal splenorenal shunting (39.3%) when compared with distal splenorenal shunting (14.8%) and with esophagogastric devascularization with splenectomy (0%). Lethality was also significantly higher after proximal splenorenal shunting (42.9%) when compared with distal splenorenal shunting (14.8%) and with esophagogastric devascularization with splenectomy (7.1%). Indirect hyperbilirubinemia was absent after esophagogastric devascularization with splenectomy and more frequent after distal splenorenal shunting (52%) although also present after proximal splenorenal shunting (29.6%). Esophagogastric devascularization with splenectomy was demonstrated to be the best option because of the absence of encephalopathy and because of low mortality rates. Hepatic encephalopathy occurred after distal splenorenal shunting but in a lesser percentage than after proximal splenorenal shunting. The higher incidence of encephalopathy and lethality proscribes proximal splenorenal shunting in Manson'schistosomiasis.

Adolescent↗

Responses of floccular Purkinje cells to sinusoidal vertical rotation and effects of muscimol infusion into the flocculus in alert cats.

To understand how the flocculus is involved in the integration process in the vertical VOR, we examined the responses of floccular Purkinje cells during whole-body rotation in many vertical and horizontal planes in alert cats. The great majority of cells that responded to pitch rotation received excitation from the contralateral posterior canal, whereas the minority received excitation from the anterior canal, either on the ipsilateral or contralateral side; 35% of these vertical canal responding cells received convergent inputs, either from the ipsilateral or contralateral horizontal canal. After infusion of a GABA agonist (muscimol) into the area where many posterior canal responding Purkinje cells were recorded, the downward VOR induced by nose-up pitch was selectively impaired at low stimulus frequencies, together with an impairment of gaze holding after downward saccades. These results indicate a crucial role for posterior canal responding floccular Purkinje cells in generating downward eye position signals. Since the flocculus inhibits VOR interneurons related to the anterior and horizontal canals but not the posterior canal (Ito, 1984), our results suggest the possibility that posterior canal responding floccular Purkinje cells may form a transfloccular inhibitory pathway to anterior canal related VOR interneurons. This pathway may facilitate the positive feedback effect in the vestibular commissural inhibitory pathway between the co-planar vertical canals for the posterior canal signal, and may thereby be specifically involved in integration in the vertical VOR.

Animals↗

Site-directed mutagenesis of Glu-141 and His-223 in Pseudomonas aeruginosa elastase: catalytic activity, processing, and protective activity of the elastase against Pseudomonas infection.

Both Pseudomonas aeruginosa elastase and Bacillus thermoproteolyticus thermolysin are zinc metalloproteases. On the basis of the high homology of the P. aeruginosa elastase with the Bacillus thermolysin, we hypothesized that Glu-141 and His-223 are the key residues for catalytic activity of the Pseudomonas elastase. To test this possibility, we replaced Glu-141 with Asp, Gln, and Gly and His-223 with Gly, Glu, and Leu by site-directed mutagenesis. These substitutions dramatically diminished the proteolytic activities of the mutant elastases when they were expressed in Escherichia coli cells. Although these mutant elastase precursors (proelastases) were produced, no appreciable processing was observed with these mutants. The possibility that autocatalysis is involved in both the processing and activation of elastase is discussed. Furthermore, by immunizing mice with vaccines made from these mutant elastase, we were able to obtain good protection against an intraperitoneal P. aeruginosa challenge.

Animals↗

[Randomized study comparing mitral valve replacement with and without preservation of mitral ring chordae tendinae papillary continuity].

PURPOSE: To demonstrate the importance of the preservation of mitral annulus-chordae tendineae-papillary muscles continuity in mitral valve replacement. METHODS: We studied 21 patients who were submitted to mitral valve replacement, divided in two randomized groups: group 1, 12 cases who undergone mitral valve replacement, with preservation of the posterior leaflet and correspondent chordae tendineae; and group 2-9 cases who undergone conventional mitral valve replacement, excising the mitral valve apparatus. The left ventricular function was studied both, in the pre and post operative period, by echocardiography, cardiac catheterization, and radioisotopic study. The statistical analysis was done by the Wilcoxson's test. RESULTS: There were no early post operative deaths. Analyzing the results of the ejection fraction by the radioisotopic study we found a significant difference (p = 0.03) between the percentual decrease of the two groups. The results of the fractional shortening were higher in group 1 than in group 2, however not significant. The left ventricular diastolic diameters average was lower in group 1 than in group 2, so as the left atrium diameter. We found a decrease in left ventricular end-diastolic pressure in group 1, however there was an increase in group 2, by the cardiac catheterization. There was a proportional increase in group 1 both in lung artery and lung capillary pressures. There was a significant difference (p = 0.05) between the average values of right ventricular diastolic pressure. CONCLUSION: There is better preservation of left ventricular function in group 1.

Adult↗

Structural gene and complete amino acid sequence of Vibrio alginolyticus collagenase.

The DNA encoding the collagenase of Vibrio alginolyticus was cloned, and its complete nucleotide sequence was determined. When the cloned gene was ligated to pUC18, the Escherichia coli expression vector, bacteria carrying the gene exhibited both collagenase antigen and collagenase activity. The open reading frame from the ATG initiation codon was 2442 bp in length for the collagenase structural gene. The amino acid sequence, deduced from the nucleotide sequence, revealed that the mature collagenase consists of 739 amino acids with an Mr of 81875. The amino acid sequences of 20 polypeptide fragments were completely identical with the deduced amino acid sequences of the collagenase gene. The amino acid composition predicted from the DNA sequence was similar to the chemically determined composition of purified collagenase reported previously. The analyses of both the DNA and amino acid sequences of the collagenase gene were rigorously performed, but we could not detect any significant sequence similarity to other collagenases.

Amino Acid Sequence↗

Ocular torsion produced by unilateral chemical inactivation of the interstitial nucleus of Cajal in chronically labyrinthectomized cats.

Unilateral lesions of the rostral midbrain involving the interstitial nucleus of Cajal (INC) have been known to produce ocular torsion in alert animals including humans, which has been assumed to be the result of an impaired otolith-ocular reflex. We examined the effects of chemical deactivation of the INC using a gamma-aminobutyric acid (GABA) agonist (muscimol) in cats that had received bilateral labyrinthectomy, and compared the results with those in normal cats. Ocular torsion with a magnitude similar to that observed in normal cats appeared in chronically labyrinthectomized cats after unilateral muscimol infusion into the INC, indicating that ocular torsion following unilateral INC deactivation can be produced by a mechanism independent of the otolith-ocular reflex.

Animals↗

The interstitial nucleus of Cajal is involved in generating downward fast eye movement in alert cats.

We examined whether or not the interstitial nucleus of Cajal (INC) is involved in generating vertical fast eye movement. We used a gamma-aminobutyric acid (GABA) agonist (muscimol) to deactivate the INC cell bodies bilaterally in alert head-fixed cats. The INC was identified by recording vertical eye position-related burst-tonic neurons. Following muscimol infusion into the bilateral INC, the cats were unable to hold eccentric upward eye position after saccades, and the mean time constant of postsaccadic drift was 0.4 sec. In addition, downward saccades were virtually lost without clear impairment of horizontal saccades. Muscimol infusion into the Forel's field H resulted in loss of both upward and downward fast eye movement as reported previously. Bilateral electrolytic INC lesions produced results similar to those after muscimol infusion into the INC, suggesting that the cutting of passing fibers in the INC alone cannot explain the selective loss of downward fast eye movement. These results indicate that the INC itself is involved in generating downward fast eye movement. We propose that the loss of activity in possible downward burster-driving neurons in the INC can explain the selective loss of downward fast eye movement after bilateral INC lesions.

Animals↗

Otolith-visual interaction in the control of eye movement produced by sinusoidal vertical linear acceleration in alert cats.

1. Eye movement responses were examined in alert cats during sinusoidal vertical linear acceleration. Stimulus frequencies of 0.20-0.85 Hz with a constant amplitude of 10.5 cm (corresponding to 0.02-0.31 g) were used. A random visual pattern was presented to give sinusoidal vertical optokinetic stimuli of similar amplitude and frequency to the up-down motion of the cat. 2. Sinusoidal linear acceleration in the presence of a stationary visual pattern produced robust eye movement responses with near compensatory phase at all stimulus frequencies tested. With both eyes covered, a vertical linear vestibulo-ocular reflex (LVOR) was frequently produced at a stimulus strength corresponding to 0.04-0.31 g. The evoked LVOR was always small, and the overall mean response phase values advanced by as much as 70 degrees at frequencies below 0.56 Hz, indicating that the otolith signals activated by sinusoidal linear acceleration were not, by themselves, converted into compensatory eye position signals under these experimental conditions. 3. Optokinetic stimulation alone produced more lag of response phase as stimulus frequency increased, and the gain of evoked eye movement responses was smaller at higher stimulus frequencies compared to the gain during linear acceleration in the light. Bilateral labyrinthectomies resulted in a significant change of the eye movement responses during linear acceleration when visual inputs were allowed: there was more phase lag at higher stimulus frequencies and a decreased gain at all frequencies tested. These results indicate that the interaction of otolith and visual inputs produces robust eye movement responses with near compensatory phase during sinusoidal linear acceleration in the light.

Acceleration↗

Activity of eye movement-related neurons in and near the interstitial nucleus of Cajal during sinusoidal vertical linear acceleration and optokinetic stimuli.

1. A total of 43 neurons that showed a close correlation with vertical eye movement with a burst-tonic or tonic type response during spontaneous saccades, were recorded within, and in the close vicinity of, the interstitial nucleus of Cajal (INC) in alert cats. Neuronal responses to sinusoidal vertical linear acceleration (0.2-0.85 Hz, amplitude 10.5 cm) and optokinetic stimuli (0.1-1.0 Hz, amplitude 10.5 cm), were examined. 2. All 43 eye movement-related neurons responded to sinusoidal vertical linear acceleration in the presence of a stationary visual pattern in correlation to robust eye movement responses with compensatory phase. Phase and gain values (re stimulus position) of response of individual cells were independent of the stimulus frequencies tested. Of these, 33 cells were examined during linear acceleration without visual input. Most cells (27/33) did not respond even when a weak linear vestibulo-ocular reflex was present (6/27). The remaining 6 cells (6/33) responded to linear acceleration. Their mean phase values advanced by 80 degrees and gain dropped by 55% compared to the responses with visual inputs. 3. Twenty eight of the 43 cells were examined during vertical optokinetic stimuli. The activity of all 28 cells was modulated in correlation to eye movement responses. Response phase showed more lag, and gain decreased as stimulus frequencies increased, similar to optokinetic eye movement responses. 4. The close correlation between the activity of eye movement-related neurons in the INC region and robust eye movements during linear acceleration with visual inputs and optokinetic stimuli suggest that these neurons are involved in some aspect of vertical eye position generation during such stimuli.

Acceleration↗

Latencies of response of eye movement-related neurons in the region of the interstitial nucleus of Cajal to electrical stimulation of the vestibular nerve in alert cats.

Recent studies have shown that the interstitial nucleus of Cajal (INC) in the midbrain reticular formation is involved in the conversion of vertical semicircular canal signals into eye position during vertical vestibulo-ocular reflexes. Secondary vestibulo-ocular relay neurons related to the vertical canals, which constitute the majority of output neurons sending signals from the vestibular nuclei directly to the oculomotor nuclei, have been shown to project axon collaterals to the region within and near the INC. To understand how the INC is involved in the signal conversion, latencies of response of neurons in the INC region to electrical stimulation of the vestibular nerve were examined in alert cats. The responses of 96 cells whose activity was clearly modulated by sinusoidal pitch rotation (at 0.31 Hz) were analyzed. These included 41 cells whose activity was closely correlated with vertical eye movement (38 burst-tonic and 3 tonic neurons), and 55 other cells (called pitch cells as previously). Twenty nine of the 96 cells (30%) were activated at disynaptic latencies following single shock stimulation of the contralateral vestibular nerve. Disynaptically activated cells were significantly more frequent for pitch cells than for eye movement-related cells (25/55 = 45% vs 4/41 = 10%; p less than 0.001, Chi-square test). Conversely, cells that did not receive short-latency activation (less than 6 ms) were more frequent among eye movement-related cells than pitch cells (26/41 = 63% vs 13/55 = 24%; p less than 0.001, Chi-square test).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

The origin of high and regular discharge rates of eye-movement-related neurons in the region of the interstitial nucleus of Cajal.

Burst-tonic neurons in the region of the interstitial nucleus of Cajal (INC) that show a close correlation to vertical eye movement have been known to exhibit high and regular discharge rates, not only during fixation in alert animals, but also during sleep. Since they receive major input from vertical semicircular canals, we examined in this study whether or not the source of the high and regular discharge rates was the primary vestibular afferents. Infusion of lidocaine into the middle ear bilaterally resulted in a significant decrease of mean discharge rates and an increase in the coefficient of variation of the mean rates. However, burst-tonic neurons in cats that had received bilateral labyrinthectomy 6 weeks previously still exhibited high and regular discharge rates similar to those of normal cats. These results indicate that high and regular discharges of eye-position-related INC cells are maintained largely by input from primary vestibular afferents in normal cats. However, such characteristic discharges could also be maintained centrally in the brainstem without peripheral vestibular input in labyrinthectomized cats.

Analysis of Variance↗