The objective of this study was to determine the expression and activity of CTLA4 in T-cells of systemic lupus erythematosus (SLE) patients. Expression of CTLA4 on freshly isolated peripheral blood T-cells was evaluated in 33 SLE patients and 25 controls using flow cytometry. The T-cells from 19 SLE patients and 22 controls were stimulated and cultured with Chinese hamster ovary cells expressing CD80 (CHO-CD80) or with CHO cells. T-cell proliferation was determined with [3H] thymidine incorporation (CPM), and the inhibitory effect of CTLA4 on T-cell proliferation was evaluated by the ratio of CPM for T-cells with CHO-CD80 cells to that of T-cells with CHO cells (the CHO-CD80/CHO ratio). Intracellular CTLA4 expression in freshly isolated peripheral blood T-cells was significantly higher in SLE patients than the controls (P < 0.05), but there was no correlation with clinical features or disease activity. The CHO-CD80/CHO ratio of SLE patients was significantly higher than that of the controls (P < 0.05). Among SLE patients, the CHO-CD80/CHO ratio of patients with lupus nephritis was significantly higher than that of patients without lupus nephritis (P < 0.05). In conclusion, our data suggest that CTLA4 expression is not impaired in SLE patients, but there is a possibility of decreased inhibitory effect of CTLA4 involved in the pathogenesis of SLE.

Adolescent↗

PubMed2003

Can ultrasound predict histological findings in regenerated cartilage?

OBJECTIVE: To evaluate regenerated articular cartilage quantitatively by introducing an ultrasonic probe into the knee joint under arthroscopy and analysing the A-mode echogram by means of wavelet transformation. METHODS: Three experimental rabbit models (spontaneous repair model, large cartilage defect model, treatment model) were examined using our ultrasonic evaluation system and a histological grading scale. From resulting wavelet map, the percentage of maximum magnitude was selected as the quantitative index of the ultrasonic evaluation system. RESULTS: The percentage maximum magnitude in the spontaneous repair model was 61.1%, that in the large defect model was 29.8% and that in the treatment model was 36.3%. There was modest correlation between the percentage maximum magnitude and the histological grading scale (r = -0.594) CONCLUSION: Our findings indicate that ultrasound analysis can predict the microstructure of regenerated cartilage.

Animals↗

PubMed2003

Chronic active VZV infection manifesting as zoster sine herpete, zoster paresis and myelopathy.

After lumbar-distribution zoster, an HTLV-1-seropositive woman developed chronic radicular sacral-distribution pain (zoster sine herpete), cervical-distribution zoster paresis and thoracic-distribution myelopathy. Detection of anti-varicella zoster virus (VZV) IgM and VZV IgG antibody in cerebrospinal fluid (CSF), with reduced serum/CSF ratios of anti-VZV IgG compared to normal serum/CSF ratios for albumin and total IgG, proved that VZV caused the protracted neurological complications. Diagnosis by antibody testing led to aggressive antiviral treatment and a favorable outcome.

Chronic Disease↗

PubMed2003

Inter--rater and test--retest reliability of the Japanese version of the subjective deficit syndrome scale.

The subjective deficit syndrome scale (SDSS) was established in 1990 by Jaeger et al. to assess systematically subjective experiences of schizophrenic patients. It was translated into Japanese in 1999 by this paper's second author (T.I.). In this study, the inter-rater reliability of this Japanese version (SDSS-J) was examined. The subjects studied were 13 schizophrenic patients (male 6, female 7; respective average ages at study entry and disease onset: 41 and 25 years), who were being followed up at the outpatients' service of the Department of Neuropsychiatry, Hyogo College of Medicine. Four psychiatrists attended together, the systematic interview with each subject to rate the SDSS-J and independently rated 19 items. The severity scales of the items assessed by these raters ranged from 0 to 4 for 12 items and from 0 to 3 for 7. The ANOVA ICC inter-rater reliability values for the 19 individual items ranged from 0.67 to 0.96. The ANOVA ICC test-retest reliability values achieved by two raters were also high overall, ranging from 0.72 to 1.00, except for one item (item 13) assessed by one rater. Our results suggest that the SDSS-J is a potentially useful rating scale for evaluating subjective experiences of schizophrenic subjects.

Adolescent↗

PubMed2003

Increase of hippocampal acetylcholine release at the onset of dark phase is suppressed in a mutant mice model of evening-type individuals.

We have previously reported that clock mutant mice on Jcl:ICR background show about 2-h delayed circadian profiles in body temperature, spontaneous activity and sleep-wake rhythm, and that they appear to be useful as a model of evening-type of individual. Hippocampal acetylcholine (ACh) release which is positively correlated with attention, learning and memory shows a circadian variation. In this study, changes in hippocampal ACh release in transitional phase from light (rest) to dark (active) period in clock mutant mice were monitored using an in vivo microdialysis method. Compared with wild mice, the increase in hippocampal ACh in the first 2 h of the active period in the mutant mice was suppressed in parallel with peak frequency in electroencephalogram theta rhythm. The molecular basis of the circadian system appears to have a strong effect on hippocampal cholinergic function, and is probably associating with individual temporal differences in voluntary behavior, cognition, learning and/or memory performance.

Acetylcholine↗

PubMed2003

Nerve growth factor attenuates hippocampal cholinergic deficits and operant learning impairment in rats with entorhinal cortex lesions.

In the present study we investigated whether continuous intraventricular nerve growth factor (NGF) infusion could ameliorate hippocampal cholinergic deficits and learning impairment following entorhinal cortex lesions. Rats with such lesions received continuous intraventricular infusions of NGF or vehicle. Unlesioned rats with a sham operation were studied as controls. After learning sessions, a dialysis probe was implanted in the CA3 hippocampal region. In addition, brain sections were stained for synaptophysin immunoreactivity. In rats undergoing surgical procedures similar to those in the behavioral study, brains were processed for acetylcholinesterase (AChE) histochemistry. NGF-treated rats showed partial amelioration of lesion-associated hippocampal acetylcholine (ACh) efflux deficits and fixed-interval schedule learning impairment compared with vehicle-treated rats. Histochemical, immunohistologic, and microdensitometric analyses confirmed greater density of AChE-positive fibers and synaptophysin immunoreactivity in CA3, in NGF-treated rats relative to vehicle-treated rats, although not as great as in sham-operation rats, indicating partial recovery in NGF-treated rats. These results suggest that enhanced performance of the learning task with NGF treatment was related to improved hippocampal cholinergic function: specifically, increased cholinergic neurotransmission resulting from NGF effects on cholinergic neurons and presynaptic terminals.

Acetylcholine↗

PubMed2002

LC-MS determination of urinary toluenediamine in workers exposed to toluenediisocyanate.

To improve the biological monitoring method for 2,6- and 2,4-toluenediisocyanate (TDI) exposure, we developed a simple and rapid method for analysis of the corresponding urinary metabolites, 2,6- and 2,4-toluenediamine (TDA) using liquid chromatograph-mass spectrometry (LC-MS). One ml of urine was hydrolyzed at 100 degrees C for 1.5 h with H(2)SO(4). Alkalinized hydrolysate was extracted with dichloromethane (DCM) and analyzed by atmospheric pressure chemical ionization (APCI) LC-MS, in positive-ion mode. The mass spectra of TDA isomers showed the protonated molecule [M+H](+), at m/z 123 as the base peak. Calibration curves of 2,6-TDA were linear up to 400 microg/l. TDA isomers in urine of exposed workers as determined by LC-MS correlated well with those obtained by gas chromatography-mass spectrometry. 2,6- and 2,4-TDA were not detected in non-exposed subjects, whereas exposed workers showed urinary levels up to 250 and 63 microg/l, respectively.

Chromatography, Liquid↗

PubMed2002

Expression of two kinds of recombinant glutamate dehydrogenase from Aeropyrum pernix with different N-terminal sequence length in Escherichia coli.

Two recombinant Aeropyrum pernix glutamate dehydrogenase (GDH) enzymes with different length N-termini were cloned and expressed in Escherichia coli: sGDH begins with the amino acid sequence of the extracted native enzyme (M-Q-P-T-D-P-L-E-E), whereas lGDH begins with the sequence of the predicted ORF (M-E-V-L-A-L-Q-P-T-D) and is longer than sGDH by five amino acids (M-E-V-L-A). Purified recombinant lGDH was more stable than sGDH, indicating that the N-terminal extension, containing mostly hydrophobic residues, affected the overall stability of recombinant lGDH. This stabilising effect of extending the N-terminal sequence on an oligomeric enzyme such as GDH is novel; factors affecting stabilisation have previously only been discussed in the context of the contribution of internal amino acids.

Amino Acid Sequence↗

PubMed2002

Long-term response to zonisamide in patients with West syndrome.

The long-term effectiveness of zonisamide (ZNS) was evaluated in 11 patients with West syndrome (7 symptomatic) who had cessation of spasms with ZNS monotherapy. During the follow-up period (24 to 79 months, mean = 53 months), this response was maintained in 7 patients (3 symptomatic, relapse rate = 36%), including 2 children in whom ZNS was successfully discontinued. No serious adverse reactions were noted. ZNS may be both effective and well tolerated for the treatment of West syndrome.

Adolescent↗

PubMed2002

Influence of sleep disturbance on steroid 5alpha-reductase mRNA levels in rat brain.

Sleep deprivation has been shown to affect the production of steroid hormones in peripheral steroidogenic organs, but little is known about the influence of sleep disturbance on the metabolism of steroid hormones in the brain. To elucidate a possible association of the sleep-wake cycle with brain neurosteroid metabolism, the influence of short-term sleep disturbance on the expression of mRNA encoding steroid 5alpha-reductase, the enzyme converting progesterone and other steroid hormones to their neuroactive 5alpha-reduced metabolites, was investigated. Rats were first subjected to non-selective disturbance of the sleep-wake cycle, and the expression of steroid 5alpha-reductase mRNA in rat hippocampus and brainstem was determined using a semi-quantitative one-step RT-PCR technique. Non-selective disturbance of the sleep-wake cycle resulted in the elevation of 5alpha-reductase mRNA levels in the brainstem, but not in the hippocampus, and the elevated mRNA expression returned to the basal levels after a short period of the sleep recovery. Further studies showed that selective REM sleep deprivation significantly elevated 5alpha-reductase mRNA levels in both hippocampus and brainstem, thus proposing the possibility that REM sleep reduction may largely contribute to the elevation of steroid 5alpha-reductase mRNA levels observed during short-term disturbance of the sleep-wake cycle. Since the enhancement of steroid 5alpha-reductase gene expression may result in the elevation of neuroactive 5alpha-reduced steroid production in the brainstem, the findings presented here provide further evidence for suggesting that neuroactive steroids may play a physiologically important role in the neuronal network for REM sleep initiation and maintenance.

3-Oxo-5-alpha-Steroid 4-Dehydrogenase↗

PubMed2002

Sevoflurane preconditions stunned myocardium in septic but not healthy isolated rat hearts.

BACKGROUND: Recent evidence indicates that sevoflurane treatment before prolonged ischaemia reduces infarct size in normal hearts, mimicking ischaemic preconditioning. We examined whether exposure to sevoflurane before brief ischaemia, inducing a 'stunned myocardium', provided such protective effects in an isolated working heart from normal or septic rats. METHODS: With institutional approval, 91 rats were randomly allocated into one of either caecal-ligation and perforation (CLP: n=50) or sham (Sham: n=41) procedure groups 24 h before the study. After determination of baseline measurements, including cardiac output (CO), myocardial oxygen consumption (mVO(2)) and cardiac efficiency (CE; CO x peak systolic pressure/mVO(2)), each isolated heart was perfused with or without 2% sevoflurane for 15 min before global ischaemia (pre-ischaemia). After 15 min ischaemia and 30 min reperfusion, all hearts were assessed for functional recovery of myocardium (post-reperfusion). RESULTS: During the pre-ischaemia period, 2% sevoflurane caused a significant reduction of CO in the CLP group compared with the Sham group. During the post-reperfusion period, both CO (16.9 vs 11.0 ml min(-1)) and CE (11.2 vs 7.7 mm Hg ml(-1) ( micro l O(2))(-1)) was higher in the sevoflurane-treated vs -untreated hearts from CLP rats, and was accompanied by lower incidence of reperfusion arrhythmia compared with control hearts (8 vs 32%). In contrast, 2% sevoflurane did not provide cardioprotective effects in normal rats. CONCLUSIONS: The current study demonstrates that pre-treatment with sevoflurane minimizes myocardial dysfunction and the incidence of reperfusion arrhythmia after brief ischaemic insults in septic hearts.

Anesthetics, Inhalation↗

PubMed2002

Accuracy and reproducibility of left ventricular function from quantitative, gated, single photon emission computed tomography using dynamic myocardial phantoms: effect of pre-reconstruction filters.

We have investigated the accuracy and reproducibility of left ventricular (LV) functions using quantitative, gated, single photon emission computed tomography (SPECT) software dependent on critical frequencies of pre-reconstruction filters. This study incorporated dynamic myocardial phantoms (myocardial and cone shapes). Gated SPECT of 8-interval sets were pre-filtered with Butterworth filters (critical frequency varying between 0.16 and 1.16 cycles/cm, order 5) and with no filter. Phantoms were repositioned, and SPECT acquisitions were repeated. As the critical frequency increased, the estimated LV volume increased to reach a plateau at the level of the critical frequency, 0.54 cycles/cm. Conversely, the values of ejection fractions, wall motion and wall thickening with different filters which used critical frequency of > or =0.39 cycles/cm remained unchanged. However, LV functions and volumes were underestimated when any pre-reconstruction filter was used. Standard deviations of LV functions after repeated measurements were unaffected by different filters with critical frequencies of > or =0.39 cycles/cm. Standard deviations of LV volume, ejection fraction, wall motion and wall thickening were <2.2 ml, <0.9%, <0.6 mm and <8.7%, respectively. Therefore, with the exception of low critical frequencies, LV functions and volumes were highly reproducible when these routine reconstruction filters were used.

Algorithms↗

PubMed2002

Validation of left ventricular function from gated single photon computed emission tomography by using a scintillator-photodiode camera: a dynamic myocardial phantom study.

A scintillator-photodiode camera is able to acquire single photon emission computed tomography (SPECT) images by using a rotating chair system. We validated the left ventricular (LV) parameters of this camera system utilizing a dynamic myocardial phantom. Gated myocardial SPECT of a dynamic myocardial phantom (Hokkaido University type; end diastolic volume (EDV), 143 ml; end systolic volume (ESV), 107 ml; ejection fraction (EF), 25%) was performed with this scintillation camera. LV parameters were calculated using pre-installed software (Mirage Myocardial Perfusion SPECT) (study 1) and the other software (QGS; Cedars-Sinai) (study 2). For comparison, SPECT from a traditional Anger camera were processed by the QGS software (study 3). The estimated volumes were similar among the three studies (EDV, 110+/-8 ml in study 1, 112+/-2 ml in study 2 and 111+/-1 ml in study 3; ESV, 86+/-8 ml in study 1, 93+/-4 ml in study 2 and 91+/-2 ml in study 3). The estimated EFs were 23+/-3%, 17+/-2%, and 18+/-1%, respectively. The calculated volume within each study was underestimated by approximately the same degree. However, each estimated EF value for each study was close to the actual value. The estimated LV function using the scintillator-photodiode camera system may be considered as a suitable alternative to the traditional Anger camera system.

Algorithms↗

PubMed2001

The first detection of a Clar's hydrocarbon, 2,6,10-tri-tert-butyltriangulene: a ground-state triplet of non-Kekulé polynuclear benzenoid hydrocarbon.

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Journal Article↗