Pitavastatin, a 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitor, reduces hippocampal damage after transient cerebral ischemia in gerbils.

Pitavastatin, a 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitor, is a potent cholesterol-lowering drug that reduces the risk of myocardial infarction and stoke. In this study, we examined its neuroprotective effects against hippocampal CA1 neuronal damage following transient cerebral ischemia in gerbils. Forebrain ischemia was induced by occlusion of bilateral common carotid arteries for 5 min. Pitavastatin, at a dose of 3, 10 or 30 mg/kg, was administered orally twice a day for 5 consecutive days and transient cerebral ischemia was induced in mice 1 h after the last treatment with pitavastatin. Histopathological observations showed that neuronal damage to the hippocampal CA1 neurons, which was observed 5 days after ischemia in animals, was prevented by pitavastatin treatment. Immunohistochemical staining for copper/zinc superoxide dismutase (SOD) and manganese SOD decreased in the hippocampal CA1 sector of gerbils 2 days after ischemia when histological neuronal destruction was not yet found, but was clearly observed in pitavastatin-treated animals. These results indicate that pitavastatin can protect dose-dependently against ischemia-induced neuronal damage and that the mechanism of the neuroprotection may be related to the preservation of SODs, especially copper/zinc-SOD. This in part explains how pitavastatin therapy, which targets free radicals, has beneficial effects against disorders including ischemic stroke.

Application of quantitative ethanol detector (QED) test kit to measure ethanol concentration in blood samples.

In this paper, the applicability of the quantitative ethanol detector (QED) test kit for screening of ethanol concentrations in blood samples was investigated. The pretreatment of blood using the sulfosalicylic acid solution and the three-way stopcock followed by membrane filtration gave satisfactory results. The ethanol concentrations in whole blood samples (n=61) determined by QED correlated well with those determined by gas chromatography; the correlation coefficient indicated 0.990. Because a high correlation coefficient (0.928) was also confirmed in trial by investigators, QED test should be highly considered for ethanol screening in forensic praxis.

[Application of Q.E.D. and Alco-Screen test kits to measurements of ethanol in forensic samples].

We have investigated the applicability of the Q.E.D. (Quantitative Ethanol Detector) and Aloco-Screen test kits for screening ethanol concentrations in forensic samples, such as hemolyzed/decomposed blood, urine and vitreous humor. Because both kits were based on enzymatic color reactions, direct application of the kits to hemoglobin-rich samples gave unsatisfactory results. The deproteinization of blood with trichloroacetic acid followed by membrane filtration overcame such problem. This procedure was also effective for pretreatment of urine and vitreous humor samples to suppress excessive color development in the Alco-Screen test. The ethanol concentrations in whole blood (n = 29), urine (n = 7) and vitreous humor (n = 6) samples determined by the Q.E.D. kit correlated well with those determined by gas chromatography; the correlation coefficients were 0.986, 0.975 and 0.993, respectively. Because of its high specificity and sensitivity to ethanol, Q.E.D. seems to be highly reliable for quantitative estimation of ethanol concentrations in forensic samples. Alco-Screen also had high sensitivity, the specificity to ethanol was relatively low; the color reaction was also observed in the presence of acetone, n-propanol, toluene, methanol, ethylene glycol, methamphetamine, diazepam and dichrovos. Therefore, if forensic samples are analyzed by the Alco-Screen, it is essential to confirm the positive results using other analytical methods.

The relationship between photosensitive temporal lobe epilepsy and eye closure activity.

We describe a 16-year-old female patient affected by photo-induced temporal lobe epilepsy. During intermittent photic stimulation she showed a photoparoxysmal response in the EEG. This case was diagnosed from clinical symptoms, single photon emission computer tomography, and EEG data. The clinical symptoms were relieved by the administration of carbamazepine. As these photoparoxysmal responses were observed not only during photic stimulation, but also when patient was closing her eyes during an eye-opening test in complete darkness, we propose the existence of an alternative pathway such as from the extraocular muscles or orbicularis oculi, or activation of cortical activity due to the change of consciousness by closing eyes in inducing photosensitive epilepsy. We describe an additional case and discuss a novel aspect of photo-induced temporal lobe epilepsy.

Electrophysiological studies in spinocerebellar ataxia type 6: a statistical approach.

In spinocerebellar ataxia type 6 (SCA6), the cerebellum is predominantly affected, but several electrophysiological studies have revealed subclinical disorders other than cerebellar lesions. We conducted statistical analyses by comparing SCA6 patients and age-matched normal controls to asses whether electrophysiological abnormalities are directly associated with SCA6 because late onset of SCA6 may involve senile changes. We performed brain stem auditory evoked potentials (BAEP), visual evoked potentials, somatosensory evoked potentials and nerve conduction studies in 10 SCA6 patients. The BAEP latencies of wave I was prolonged and compound muscle action potentials of peroneal nerve and sensory nerve action potentials of sural nerve reduced in SCA6 patients. Our results suggest an existence of peripheral impairment in the auditory pathway and axonal neuropathy in SCA6.

Habituation of event-related potentials in patients with Parkinson's disease.

Auditory event-related potential (ERP) was studied in idiopathic Parkinson's disease (PD) using our new procedures. We examined 12 non-demented patients with PD, and 9 age-matched control subjects. Ninety responses induced by rare stimulation were continuously recorded from Fz, Cz, Pz referred to linked earlobe electrode (A1A2), and were divided into nine blocks (one block = 10 responses) for statistical analysis. We assessed the habituation of auditory ERP to detect delicate changes related to the information processing in PD. N100 and P300 latencies were significantly longer in PD than in control subjects (p<0.05). P300 latency gradually increased with progression of recording blocks in both PD and control subjects, whereas N100 latency increased only in PD. Three way analysis of variance for P300 amplitude revealed significant effects of subject group, recording electrode site, and trial block. P300 amplitude was smaller in PD than in control subjects. Significant negative correlation between P300 amplitudes and block numbers were observed at Cz and Pz in PD, and at Fz in the control group. Previous reports as well as present results suggest that prolongation of N100 latency might be related to frontal lobe dysfunction, and abnormality of P300 to dysfunction in both the frontal lobe and hippocampus in PD.

Adult leigh syndrome: treatment with intravenous soybean oil for acute central respiratory failure.

This study reports a 38-year-old woman with adult Leigh syndrome associated with partial deficiency of the pyruvate dehydrogenase complex. The patient had intermittent diplopia, loss of vision, dystonia, central respiratory failure and unconsciousness with lactic acidosis. Treatment with an intravenous ketogenic emulsion resulted in rapid clinical and biochemical improvement. In patients with acute respiratory failure under these circumstances, intravenous ketogenic emulsion therapy is worth consideration.

[Electrophysiological findings in patients with SCA6].

The major symptom of spinocerebellar ataxia type 6 (SCA6) is progressive cerebellar ataxia. MRI revealed isolated cerebellar atrophy without brainstem and cerebral involvement. Up to the present electrophysiological abnormalities in patients with SCA6 have not been intensively investigated. We performed electrophysiological examination, such as multi-modality evoked potentials, in 10 patients with SCA6. We analyzed the electrophysiological data including the results previously reported. When compared with SCA1, 2 and MJD, specific findings in electrophysiological studies are obscure in SCA6. Existence of subclinical lesions in peripheral nerves, pyramidal tract, auditory pathway, visual pathway, and sensory pathway was suggested in some cases with SCA6. It is important to consider the effect of age, because age at onset is relatively late in SCA6.

Spinocerebellar ataxia type 6 in relation to CAG repeat length.

UNLABELLED: The purpose of the present study was to assess the relationship between clinical characteristics of spinocerebellar ataxia type 6 (SCA6) and CAG repeat length. MATERIALS AND METHODS: We examined clinical symptoms of 54 patients with SCA6. CAG repeat length was compared among subgroups divided by clinical manifestations. RESULTS: The major symptom was progressive cerebellar ataxia. Truncal or limb ataxia, dysarthria, and nystagmus were observed in more than 80% of the patients. In analysis of CAG repeat length in patients with different types of nystagmus, CAG repeat length was the longest when both upbeat and downbeat nystagmus existed (P < 0.01). In addition, CAG repeat length was longer when the initial symptom was ataxic gait and was shorter when the initial symptom was dysarthria or ocular symptom (P < 0.05). CONCLUSION: Clinical features of SCA6 might be influenced by the length of abnormal CAG repeat.

[A case of impending neuroleptic malignant syndrome associated with Shy-Drager syndrome].

We report a 55-year-old man with impending neuroleptic malignant syndrome who showed a remarkable dysautonomia such as dysuria and was treated with L-dopa under the diagnosis of Shy-Drager syndrome. The patient demonstrated fever, leukocytosis and elevated serum creatine kinase by a decrease in L-dopa dose. Probably, he developed impending neuroleptic malignant syndrome, induced by urinary tract infection as well as decrease in L-dopa dose. Since Shy-Drager syndrome is often treated with antiparkinsonian drugs, neuroleptic malignant syndrome can possibly develop after the change in dosage of catecholaminergic drugs. The imbalance of neurotransmitters and receptors in the central autonomic nervous system may participate in the development of neuroleptic malignant syndrome. Accordingly, Shy-Drager syndrome can easily be associated with neuroleptic malignant syndrome because of its severe disturbance in the autonomic nervous system. However, autonomic nervous dysfunction, a major sign of neuroleptic malignant syndrome, might be masked by symptoms of Shy-Drager syndrome. Therefore, diagnosis of neuroleptic malignant syndrome is often delayed. Careful observations of patients with Shy-Drager syndrome related with an antiparkinsonian drug are necessary, especially when the dose of drugs is changed or the general condition deteriorates.

[A case of polyneuropathy associated with osteosclerotic myeloma].

A 50-year-old woman presented with sensorimotor polyneuropathy in the lower limbs. Biopsy of the muscle revealed neurogenic changes, and electrophysiological study and pathology of the biopsied nerve indicated myelinoaxonal degeneration. Her symptoms responded to neither plasmapheresis nor prednisolone. Pelvic radiography disclosed an osteosclerotic lesion that was subsequently diagnosed as IgA-lambda plasmacytoma by biopsy. Radiation therapy for this solitary osteosclerotic myeloma improved the patient's polyneuropathy. In Japan, reports of cases of polyneuropathy associated with osteosclerotic myeloma without multiorgan disorders as in our case have not previously been described; thus, such cases may be misdiagnosed. Therefore, in patients with polyneuropathy of unknown etiology, a systemic bone survey and repeated searches for M-protein are important. Treatment of bone lesions in patients with solitary osteosclerotic myeloma may improve the associated polyneuropathy.

The anticodon loop is a major identity determinant of Saccharomyces cerevisiae tRNA(Leu).

The recognition of tRNALeu, one of the class II tRNAs having a long variable arm, by leucyl-tRNA synthetase in Saccharomyces cerevisiae was studied using the T7 transcription system. Exchanging the anticodon arm of tRNALeu but not the D- or T psi C-arm to that of tRNASer seriously affected the leucine accepting activity. Two nucleotides in the anticodon loop, A35 and G37, were found to be important for leucylation. It was also found that the discriminator base, A73, is required for leucylation, and G73 of tRNASer functions as a negative identity determinant for leucyl-tRNA synthetase. Introducing a set of three base substitutions at positions 35, 37 and 73 was sufficient to convert tRNASer into an efficient leucine acceptor. These results indicate that the identity elements of tRNALeu lie at the second position of the anticodon and the 3' adjacent to the anticodon as well as the discriminator position. Such a sequence specific recognition manner is significantly different from that of Escherichia coli, in which not the anticodon but the tertiary structural elements play a key role in discriminating from other class II tRNAs. The leucine system is the first example which shows that the requirement of the anticodon sequence is variable among species.

Analysis of hemoglobin in bloodstains using high-performance liquid chromatography.

Hemoglobin (Hb) extracted from bloodstains was investigated using cation-exchange high-performance liquid chromatography (HPLC). Among the HbA1 fractions, the amount of HbA1d increased linearly as the ages of the bloodstains increased, although the amount of HbA1c remained virtually unchanged. As HbA1 fractions are Hb modified with glucose or glycolytic intermediates, the total Hb were separated into glycated and non-glycated Hb fractions using affinity chromatography to investigate the effects of glucose on the age-induced Hb changes. The amounts of glycated Hb increased in proportion to the blood glucose level, and the increased glycated Hbs were eluted at retention times that corresponded to those of HbA1d and HbA0, although HbA1c peak area was virtually unchanged. The amounts of both glycated and non-glycated HbA1d increased with bloodstain aging, and the proportion of non-glycated HbA1d relative to the total HbA1d was greater than that of glycated HbA1d. These findings show that glucose has little effects on the elevation of HbA1d in bloodstains that occurs as they age. Furthermore, measuring HbA1d levels in bloodstains using HPLC may prove useful for the determination of their ages.

Steroid hormone production in testis, ovary, and adrenal gland of immature rats irradiated in utero with 60Co.

Pregnant rats received whole-body irradiation at 20 days of gestation with 2.6 Gy lambda rays from a 60Co source. Endocrinological effects before maturation were studied using testes and adrenal glands obtained from male offspring and ovaries from female offspring irradiated in utero. Seminiferous tubules of the irradiated male offspring were remarkably atrophied with free germinal epithelium and containing only Sertoli cells. Female offspring also had atrophied ovaries. Testicular tissue obtained from intact and 60Co-irradiated rats was incubated with 14C-labeled pregnenolone, progesterone, 17 alpha-hydroxyprogesterone, and androstenedione as a substrate. Intermediates for androgen production and catabolic metabolites were isolated after the incubation. The amounts of these metabolites produced by the irradiated testes were low in comparison with the control. The activities of delta 5-3 beta-hydroxysteroid dehydrogenase, 17 alpha-hydroxylase, C17,20-lyase, and delta 4-5 alpha-reductase in the irradiated testes were 30-40% of those in nonirradiated testes. Also, the activities of 17 beta- and 20 alpha-hydroxysteroid dehydrogenases were 72 and 52% of the control, respectively. In adrenal glands, the 21-hydroxylase activity of the irradiated animals was 38% of the control, but the delta 5-3 beta-hydroxysteroid dehydrogenase activity was comparable to that of the control. On the other hand, the activity of delta 5-3 beta-hydroxysteroid dehydrogenase of the irradiated ovary was only 19% of the control. These results suggest that 60Co irradiation of the fetus in utero markedly affects the production of steroid hormones in testes, ovaries, and adrenal glands after birth.

Glutamate-pyruvate transaminase (GPT) variant found in a disputed paternity case.

GPT variants, GPT2C-1 and GPT2A-2C, were found in samples from a mother and her child, respectively, in a disputed paternity case. The phenotype GPT 2C-1 was determined by polyacrylamide gel electrophoresis performed at alkaline pH and acidic pH; this is probably the first GPT*2C-1 detected in Japanese. It was considered that the GPT 2C allele in this family was inherited in a manner of autosomal codominant transmission.

A case of a newborn infant with Hb M Iwate.

Hemoglobin of a newborn infant who was suspected to hereditarily have Hb M Iwate was examined. The infant hemolysate was separated into five fractions by column chromatography on Amberlite CG-50, and two of these fractions showed absorption spectra corresponding with that of Hb M Iwate. Five bands were found after the isoelectric focusing of the hemolysate, and two of these bands were brown. The two Hb M fractions obtained by column chromatography was focused to the positions of the brown bands. One of these Hbs M corresponded with Hb M Iwate (alpha M2 beta 2) from an adult carrier of this trait, but the other was not found in adult hemolysates. The latter species of Hb M was shown to be composed of the abnormal alpha chain and the normal gamma chain (alpha M2 gamma 2) by chain analysis, and was assumed to be specific for infants. A quantitative estimation of the hemoglobins in the infant hemolysate showed that there was no difference between the relative quantities of the fetal and adult forms of Hb M Iwate.