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A microwave (MW)-assisted method for the continuous production of hydrogen from methylcyclohexane using platinum on spherical carbon beads has been developed, and its application to the efficient dehydrogenative aromatization of fully saturated cyclic alkanes, including piperidines as representative heterocycles, has been studied. Effective dehydrogenation was achieved by the construction of a highly intense energy field, which acted as a reactive site, on the irradiation of the carbon beads (CB) support. The reaction could be carried out with only 10-W single-frequency MWs. The catalyst system could be used continuously for at least 12 h without any loss of catalyst activity. Dehydrogenative aromatization could also be catalyzed and simple cyclohexane derivatives, as well as piperidine derivatives as representative N-heterocyclic alkanes, were tested.

This chapter presents examples of and advocates for the adoption of tunable solid-state (semiconductor) oscillator single-mode microwave flow reactors toward laboratory and larger-scale synthetic chemistry applications. Tunable solid-state oscillator single-mode microwave flow reactors are more versatile heaters that impart both better process control and energy efficiency than conventional magnetron oscillator flow reactors when operated in single-mode or multimode.

Microwave heating in chemical reactions was first reported in 1986. There have since been many reports employing microwave heating in organic chemistry, where microwave heating has afforded higher yields of products in shorter time periods. However, such reactions are challenging to scale in batch due to the limited penetration depth of microwaves as well as the wave propagation dependence on cavity size. Continuous flow has addressed both these issues, enabling scalability of microwave processes. As such, a host of reports employing microwave flow chemistry have emerged, employing various microwave heating and reactor configurations in the context of either custom-built or commercial apparatus. The focus of this review is to present the benefits of microwave heating in the context of continuous flow and to characterize the different types of microwave flow apparatus by their design (oscillator, cavity type and reactor vessel). We advocate the adoption of tunable, solid-state oscillator single-mode microwave flow reactors which are more versatile heaters, impart better process control and energy efficiency toward laboratory and larger-scale synthetic chemistry applications.

We have developed a single-mode microwave reactor for continuous-flow synthesis with various methods of operation. This device measures the resonant frequency and modulates the oscillation frequency accordingly to maintain the maximum electric field intensity in the cavity. It can be operated either using constant applied irradiation power or using new programs which change the electric power (E-GRA) and change the flow rate (Fl-GRA), in order to rapidly screen various reaction conditions. As a model reaction, the Claisen rearrangement reaction of allyl 1-naphthyl ether 1 was rapidly optimized in this device, affording the desired product 2-allylnapthalen-1-ol 2 in high (91%) yield and up to 20 g/h productivity.

We demonstrated the rapid Johnson–Claisen rearrangement of allyl alcohol and triethyl orthoacetate with a continuous flow apparatus combined with a microwave reactor. The reaction could be carried out without solvent, and only a catalytic amount of acetic acid was sufficient to promote the reaction under microwave irradiation conditions. To confirm the optimal reaction conditions found experimentally, we performed Design of Experiments (DoE) by the Nelder–Mead method and a least-squares method regarding the amount of acetic acid and the flow rate. Consequently, the highest yield of the desired γ,δ-unsaturated ester was obtained, and the productivity at the reaction step of the continuous process was 89.5 g/h under the optimal conditions, suggesting that 2.1 kg of the product would be theoretically obtained in 1 day. We also investigated the Johnson–Claisen rearrangement using other allylic alcohols, and the corresponding products were obtained in good to high yields per unit of time.

Background: Although diagnosis of isolated acute vestibular syndrome (AVS) via physical examination has been reported, a great deal of experience is needed to interpret the results. Patients with AVS often cannot participate in a physical examination, and not every emergency department (ED) is equipped with a magnetic resonance imaging machine. Therefore, in isolated AVS, clinical factors other than those used by general physicians and/or residents are required to identify strokes in these patients. The aim of this study was to determine those clinical factors useful in differentiating stroke patients from isolated AVS patients in the ED setting. Methods: We examined the records of 116 consecutive patients who presented to the ED between May 2010 and April 2013 with AVS (acute vertigo or dizziness, nystagmus, nausea/vomiting, head-motion intolerance, unsteady gait) and without auditory symptoms, head and/or neck pain, or neurological findings. Patients underwent diagnostic neuroimaging. Clinical characteristics and vital signs measured in the ambulance were assessed. Multivariate logistic regression analysis was performed to evaluate the associations of systolic blood pressure (SBP) ≥150 mmHg, diastolic blood pressure (DBP) ≥85 mmHg, heart rate, age ≥60 years, and dyslipidemia with stroke. Results: Stroke was diagnosed in eight of the 116 (6.9%) patients with isolated AVS. DBP ≥85 mmHg (odds ratio 29.0; 95% confidence interval 3.38–249) was the only significant and independent factor associated with stroke among patients with isolated AVS. Conclusion: This study identified DBP ≥85 mmHg to be a simple and useful clinical factor differentiating stroke from non-stroke isolated AVS.

AIM Atrial fibrillation (AF)-related cardioembolic stroke is a serious problem in the aging society. The present study examined the clinical characteristics and outcomes of AF-related cardioembolic stroke in patients aged ≥80 years. METHODS Between September 2011 and April 2014, consecutive patients with ischemic stroke and AF were retrospectively extracted from the multicenter database. Clinical characteristics were compared between patients aged ≥80 years and

We report the case of a 68-year-old man who suddenly developed right hemiparesis. MRI of the brain revealed a new infarction in the left corona radiata, and intravenous rt-PA was administered 100 minutes after the onset of symptoms. After the rt-PA infusion was started, his consciousness declined and the CT revealed bilateral intracerebral hemorrhage. His consciousness did not improve thereafter. He was diagnosed with ANCA-associated vasculitis owing to the high levels of MPO-ANCA and urinary protein and the presence of erythrocytes without any other cause of renal damage. Although ANCA-associated vasculitis rarely involves the central nervous system, intracerebral hemorrhage as well as cerebral infarction may occur. This case provides an important opportunity to explore the implications of intravenous rt-PA therapy for acute cerebral infarction in patients with ANCA-associated vasculitis.

Background We examined the association between pre-admission risk scores and severity on admission and functional outcome in acute ischemic stroke with atrial fibrillation (AF). Methods Between September 2011 and April 2014, we retrospectively extracted consecutive ischemic stroke patients with AF whose pre-admission modified Rankin Scale (mRS) score was 2 or less from our prospective database. Pre-admission CHADS 2 , CHA 2 DS 2 -VASc, and R 2 CHADS 2 scores were calculated in each patient, and their association with the National Institutes of Health Stroke Scale (NIHSS) score on admission or unfavorable outcome (mRS ≥3 at 3 months from the onset) was assessed. Results A total of 344 patients (189 were men; age, 77.7 ± 10.0 years) were included in the analysis. The median pre-admission CHADS 2 , CHA 2 DS 2 -VASc, and R 2 CHADS 2 scores were 2, 4, and 4, respectively. NIHSS score on admission was positively correlated with pre-admission CHADS 2 (ρ = .116, P = .031), CHA 2 DS 2 -VASc (ρ = .166, P = .020), and R 2 CHADS 2 scores (ρ = .106, P = .049). Receiver operating characteristic (ROC) curve analysis revealed that pre-admission CHADS 2 score of 2 or more (sensitivity, 80%; specificity, 45%; area under the ROC curve [AUC], .654), CHA 2 DS 2 -VASc score of 3 or more (sensitivity, 86%; specificity, 44%; AUC, .683), and R 2 CHADS 2 score of 4 or more (sensitivity, 61%; specificity, 62%; AUC, .657) were associated with unfavorable outcome. The pre-admission CHA 2 DS 2 -VASc score was better than the pre-admission CHADS 2 score in estimating unfavorable outcome ( P = .017). In multivariate analysis, cutoffs of these scores, female sex, higher NIHSS score, and internal carotid artery occlusion were associated with unfavorable outcome. Conclusions Pre-admission CHADS 2 , CHA 2 DS 2 -VASc, and R 2 CHADS 2 scores were associated with onset severity and functional outcome in acute ischemic stroke with AF.

The Rolling Circle Amplification (RCA) is an enzymatic synthesis method of deoxyribonucleic acid (DNA) strands with repeated sequence of a circulate template-DNA. Microwave heating technology is applied to organic and inorganic chemistry to produce useful effects such as rapid heating, selective heating, decreased reaction times, and improved product yield. However, the mechanisms underlying the thermal or non-thermal effects, i.e., reaction-promoting effects, of MW chemistry are still unclear. We focus on the molecular perspective to investigate the effect of MWs via a DNA amplification reaction that utilizes enzymes. The microwave reaction temperature was strictly controlled using a microwave applicator optimized for enzymatic-scale reactions. We show that microwave heating facilitated the synthesis of repetitive DNA through RCA using the four DNA polymerases. Analysis of the temperature profiles of each RCA component subjected to microwave heating revealed the selectivity heating of buffer components compared with primers, template DNA, dNTP, and RNase-free water. Further, we showed the relation between the microwave power loss and RCA components via dielectric measurements, cavity resonator feature measurement, and electromagnetic simulation. The buffer containing ions of the RCA components was selectively heated via microwave irradiation in the TM010 cavity resonator.

The word “omics” indicates the study of a total information in biological cell, such as the genome, which is all DNA in a living cell, or the proteome which is all the proteins. Omics technologies of genomics and proteomics, such as tailor-made treatment, it has been positioned in the center of the future of medical technology. The efficiency of this technology is also expected by microwave technology. Microwave technology, for the time shortening of omics pretreatment reaction, has been fully achieved. The corresponding to the small-scale is also strongly desired to high-throughput technology such as a well plate. The corresponding to the small-scale is strongly desired to high-throughput technology such as a well plate. In this study, we tried the microwave assisted proteomics for enzymatic hydrolysis of protein molecules with a protease tripsin. Chicken egg white-derived lysozyme having a molecular weight 14,300 as a model protein was used in the enzymatic hydrolysis. The molar ratio of trypsin and lysozyme was adjusted to 1 ∶ 50, reductive alkylation lysozyme was digested with 37°C, and analyzed by MALDI-TOF MS after the reaction. An irradiation reactor of cavity resonator type was applied to enzyme reaction microwaves continuously irradiated with microwaves of 16 W, and maintained at 37°C, of the reaction temperature. The conventional method was used with a water bath. As a result, reaction time was reduced from 20 hours to 10 minutes. It was found that the reaction had proceeded completely, from Mass spectrum. In the enzymatic hydrolysis, there is the problems to digest for oneself. However, the autolysis of trypsin under microwave irradiation was not detected. Next, we had checked by MASCOT searching after TOFMS analysis, it was confirmed which part of the protein is cleaved from the protein fragments. Enzyme-substrate concentration, time of 1 to 10, the sequence coverage of 84% by microwave irradiation was 57% under normal conditions. Trypsin recognized amino acid sequence, which means that 84% is broken, it was considered to cleave amino acid residues inside the protein.

Background: We aimed to determine the predictive factors for excellent or extremely poor functional outcome in patients with first-ever atrial fibrillation (AF)-related cardioembolic stroke. Methods: Retrospective observational study from a database. Patients with AF-related cardioembolic stroke with a premorbid modified Rankin Scale (mRS) score of 0 or 1 and without a previous history of stroke were included. Results: Factors associated with excellent functional outcome (mRS scores of 0 or 1; n = 77; 30.4% of patients) included age >78 years (OR 0.31, 95% CI 0.15-0.61), male sex (OR 2.16, 95% CI 1.04-4.60), absence of hypertension (OR 0.46, 95% CI 0.22-0.94) and initial National Institutes of Health Stroke Scale (NIHSS) score of >9 (OR 0.08, 95% CI 0.03-0.16). Factors associated with extremely poor functional outcome (mRS scores of 5 or 6; n = 63; 24.9%) included age >78 years (OR 3.30, 95% CI 1.54-7.39), initial NIHSS score of >9 (OR 12.38, 95% CI 5.40-32.56), congestive heart failure (OR 4.82, 95% CI 2.00-12.19) and ischemic heart disease (OR 4.02, 95% CI 1.18-14.69). Conclusions: Predictive factors exist to delineate excellent and extremely poor functional outcomes after a first-time stroke associated with AF.

Atrial fibrillation (AF)-related cardioembolic stroke is a serious problem in the aging society. The present study examined the clinical characteristics and outcomes of AF-related cardioembolic stroke in patients aged ≥80 years.Between September 2011 and April 2014, consecutive patients with ischemic stroke and AF were retrospectively extracted from the multicenter database. Clinical characteristics were compared between patients aged ≥80 years and80 years. Multivariate cox proportional hazard models were used to estimate hazard ratios and 95% confidential intervals on 90-day mortality for age of ≥80 years.A total of 253 patients aged ≥80 years (87 men, 86.4 ± 5.0 years) and 196 patients aged80 years (134 men, 70.4 ± 7.1 years) were included. Patients aged ≥80 years were more frequently female, and more likely to have higher premorbid modified Rankin Scale score (mRS), lower body mass index, previous history of stroke, prior antiplatelet therapy, congestive heart failure, and persistent AF. Patients aged ≥80 years had higher initial National Institutes of Health Stroke Scale score and were more likely to have occlusion of the cervicocephalic arteries, but less likely to receive thrombolysis. Patients aged ≥80 years had a higher mRS and mortality after 3 months. Age of ≥80 years was a significant predictor of 90-day mortality after adjustment for sex (hazard ratio 2.20, 95% confidential interval 1.25-4.09), but was no longer significant after further adjustment for other clinical characteristics and stroke severity.In AF-related cardioembolic stroke, patients aged ≥80 years had different clinical characteristics and poorer outcome compared with patients aged80 years. Geriatr Gerontol Int 2017; 17: 708-713.

症例は40歳，男性で，頭痛，嘔吐を主訴に救急搬送された．性転換術による精巣摘出の既往があり，来院1カ月前から女性化目的に経口避妊薬を内服していた．来院時の神経学的所見では，軽度の意識障害のみで局所徴候はなかった．血液検査では凝固系の亢進があり，頭部CTでは直静脈洞，上矢状静脈洞，左横静脈洞に高吸収域を認めた．MRVでは左横静脈洞の信号消失と右横静脈洞の信号低下があり，脳静脈洞血栓症と診断した．抗凝固療法を開始し，経過中に出血性梗塞を来したものの，再開通を得た．静脈血栓症の原因として経口避妊薬は知られているが，今回はさらにアンドロゲン低下による線維素溶解能の低下が素地にあったものと考えられた．

A 35-year-old woman developed recurrent aseptic meningitis three times over a period of 16 months. Each episode followed swelling of her cervical lymph nodes. During the third episode, microscopic findings of biopsied specimens from a cervical lymph node indicated subacute necrotizing lymphadenitis (SNL). While she responded poorly to NSAIDs, steroids rapidly improved her fever, headache and swollen lymph nodes. Since the first episode, anti-nuclear antibody (ANA) and anti-SS-A antibody was positive and the titer of ANA increased with each episode. SNL is a benign and self-limited disease, and the appearance of autoantibodies is usually transient. It is possible that a persistent immune abnormality is related to recurrences of aseptic meningitis with SNL.

We present a 71-year-old woman with hereditary hemorrhagic telangiectasia (HHT) who at age 69, had undergone total gastrectomy because of repeated upper gastrointestinal bleeding. A day prior to admission she began to demonstrate abnormal behavior. Examination showed she was restless and had higher brain dysfunction. Triphasic waves were seen on EEG, and a high signal in the globus pallidus on T1-weighted MRI. Plasma NH3 level was increased after a meal. Abdominal CT scan showed vascular anomalies including a portohepatic vein shunt. She was diagnosed with portosystemic encephalopathy. After treatment with a low-protein diet, lactitol, and branched chain-amino acids, her clinical condition, plasma NH3 level after a meal, and EEG returned to normal. Because portosystemic shunt is rare in HHT, there have been few reports of portosystemic encephalopathy with this condition. However, with aging, the possibility of portosystemic encephalopathy increases because of age-related increases in portosystemic shunt volume.

Background Pure dysarthria (PD) and dysarthria-facial paresis syndrome (DFP) mainly result from lenticulostriate artery territory infarction. PD and DFP are rare clinical entities, often grouped without distinction. The purpose of this study was to examine clinical and radiographic differences between PD and DFP due to unilateral internal capsule and/or corona radiata infarction. Methods Using a database that included consecutive patients with ischemic stroke admitted to the neurological stroke units of three hospitals within 7 days from onset between September 2011 and April 2014, we retrospectively extracted first-ever stroke patient data, who presented with PD or DFP with a single ischemic lesion localized in the internal capsule and/or corona radiata. Patients with weakness, ataxia, sensory deficit, or cortical symptoms were excluded. Ischemic lesion volume was calculated by the ABC/2 method on diffusion-weighted imaging (DWI). DWI images were normalized and superimposed to the template for PD and DFP. We compared patients' characteristics between PD and DFP. Results A total of 2126 patients, including 65 patients (3.1 %) with PD or DFP, were registered. Of these, 13 PD patients and 18 patients with DFP due to unilateral internal capsule and/or corona radiata infarction were included for analysis. Compared with DFP patients, PD patients had longer onset-to-door time (median 37.5 vs. 10.8 h, p = 0.031), shorter vertical length (C component) of ischemic lesions (median 12.0 vs. 18.8 mm, p = 0.007), and smaller ischemic lesion volume (median 285 vs. 828 mm3, p = 0.023). Ischemic lesions causing PD were located more frequently in the left hemisphere than DFP (92 % vs. 56 %, p = 0.045). The superimposed lesion pattern indicated that DFP had lesions more medial and involving posterior portions of the putamen and the caudate body, as well as more of the genu and posterior limb of the internal capsule, than PD. Ninety days after onset, symptoms disappeared in 21 (72 %) out of 29 patients. Conclusions In cerebral infarction limited to the internal capsule and/or corona radiata, PD is derived from smaller and left-sided lesions with delay in diagnosis compared with DFP. The clinical course of those with PD and DFP might be benign.

Background and Purpose: Pure dysarthria (PD) and dysarthria-facial paresis syndrome (DFP) are considered as rare clinical entities of dysarthria-clumsy hand syndrome, mainly due to small vessel disease. We aimed to clarify the clinical and radiographic difference between patients with PD and DFP, especially due to internal capsule and/or corona radiata infarction, using the database of wide area network-based clinical research. Methods: This multicenter, prospective, observational study included consecutive patients with acute ischemic stroke admitted to 4 stroke centers in Fukuoka prefecture within 7 days from the onset. We included patients with first ever stroke and presenting PD or DFP with a single ischemic lesion localized in the internal capsule and/or corona radiata. Patients with any motor-sensory disturbance or cortical symptoms were excluded. Ischemic lesion volume was calculated by the A*B*C/2 method on diffusion weighted imaging. The local ethics committee approved the collection and submission of each patient’s clinical data to the study office in Kyushu University. Results: Between October 2010 and April 2014, a total of 2348 patients including 1773 (1006 men, 73.3±13.1 years) with first ever stroke were registered. Of them, 12 (0.68%) patients with PD and 18 (1.02%) patients with DFP were included for the analysis. Compared to patients with PD, patients with DFP had shorter onset-to-door time (median 10.75 [7.25-27.125] hours vs. 33.75 [14.25-51.5] hours, p = 0.042), larger vertical length (C component) of ischemic lesions (median 18.75 [12-20.625] mm vs. 9.25 [6-13] mm, p = 0.008), and greater ischemic lesion volume (median 828 [291-1664] mm3 vs. 289 [120-416] mm3, p = 0.031). Ischemic lesions causing PD were located more frequently in the left hemisphere than those of DFP (92% vs. 56%, p = 0.049). There was no difference in age, sex, and vascular risk factors between patients with PD and DFP. Symptom progression after admission was seen in one PD case which worsened to DFP. Conclusions: In cerebral infarction of the internal capsule and/or corona radiata, DFP might be derived from larger, vertically longer ischemic lesions comparing to PD. There might be stronger association between PD and left-sided lesions.

Connexin32 (Cx32) is a gap junction protein and its mutations are responsible for X-linked Charcot-Marie-Tooth disease. We examined the functional abnormality of C6 glioma cells transfected with mutant (C53S and P172R) Cx32 genes. Nontransfected C6 did not express Cx32. Northern and Western blot analyses showed Cx32 mRNA and protein in cells with the wild-type gene as well as with the mutant Cx32 genes. An immunocytochemical study of cells with the wild-type gene showed the immunoreactive spots in the cell membrane. In cells with C53S or P172R mutant gene, however, the immunoreactivity was found in the cytoplasm. The scrape-loading method produced effective dye transfer in cells with the wild-type gene but not in those with mutant genes. A cell proliferation assay showed no differences in nontransfected cells, cells with the wild-type gene and those with the mutant genes. Messenger RNA expression for proteolipid protein did not change. These findings suggest that Cx32 gene mutation results in loss of cell-to-cell communication because of failure to incorporate Cx32 protein in the cell membrane. The mutations do not, however, interfere with cell proliferation or myelin-specific gene expression, at least myelin proteolipid protein expression in C6 glioma cells. J. Neurosci. Res. 51:154–161, 1998. © 1998 Wiley-Liss, Inc.

Japanese hereditary neuropathy with liability to pressure palsy (HNPP) patients have a deletion of one peripheral myelin protein-22 (PMP22) gene region in distal chromosome band 17p11.2 as do Caucasian patients. Japanese and Asiatic Indian CMT1A patients have a PMP22 gene duplication that results in Charcot-Marie-Tooth disease type IA (CMT1A; HMSNIA) in patients of European and Middle Eastern ancestry. About 70% of Japanese CMT1 patients have a PMP22 duplication as do Caucasians, while Japanese CMT1B, CMT2 and Dejerine-Sottas patients do not have PMP22 gene region aneuploidy. Although HNPP and CMT1A genotypes are generated simultaneously by unequal recombination that results in PMP22 gene aneuploidy in each daughter cell, only 3 Japanese HNPP probands with PMP22 deletion from a large patient population were referred to a single center compared to 18 referred CMT1A probands with PMP22 duplication. This lower HNPP frequency more likely reflects lower HNPP reproductive fitness than patient ascertainment bias because disease severity and variation in severity is about the same in CMT1A and HNPP patients and because all patients of both types were referred regardless of disease severity. These results, along with an apparently high de novo CMT1A mutation rate, suggest that common ancestors of Japanese, Asian Indians, and Caucasians carried PMP22 geneflanking sequences that enhance unequal crossing over. © 1995 Wiley-Liss, Inc.

A 73-year-old man was admitted for evaluation of sudden onset of dizziness, bilateral shoulder pain, and brachial diplegia. Neurological examination revealed severe bilateral weakness of the triceps brachii, wrist flexor, and wrist extensor muscles. There was no paresis of the lower limbs. His gait was ataxic. Pinprick and temperature sensations were diminished at the bilateral C6-C8 dermatomes. Vibration and position senses were intact. An MRI of the head revealed a right cerebellar infarction and occlusion of the right vertebral artery. An MRI of the cervical spine on T₂ weighted imaging (T₂WI) showed cord compression at the C3/4-C5/6 level secondary to spondylotic degeneration without any intramedullary signal changes of the cord. On the following day, however, high-signal lesions on T₂WI appeared in the C5-C6 spinal cord, suggesting cord infarction. Unilateral vertebral artery occlusion does not usually result in cervical cord infarction because of anastomosis of arteries. Because of the long-term mechanical compression in our case, it was likely that cervical cord ischemia was present before the onset of symptoms. On the basis of chronic cord compression, our case suggests that occlusion of a unilateral vertebral artery could cause cervical cord infarction.

The repetitive passages of a Schwann cell culture results in the appearance of immortalized cells. In order to investigate the direct effects of cyclic AMP (cAMP) on Schwann cell proliferation, we used the immortalized Schwann cells because the responses of a short-term Schwann cell culture to agents increasing the intracellular cAMP are more complicated and it does not seem that all of them are due to the direct effects of cAMP. By adding up to 200 microM of forskolin, an adenylate cyclase activator, to the culture medium, Schwann cell proliferation was inhibited and the intracellular 1,2-diacylglycerol (DG) level was decreased in a dose-dependent manner to 44 and 53% of the control values, respectively. The protein phosphorylation activity in the cytosol from the cell treated with 100 microM forskolin, assayed with myelin basic protein as the acceptor, decreased to 78% and this inhibition was then reversed by the addition of 1-oleoyl-2-acetyl-sn-glycerol (OAG), a membrane-permeable DG, to the assay mixture. The cell proliferation inhibited by forskolin was also restored by the addition of OAG. These data suggest that cAMP inhibits both the activity of protein kinase C (PKC) and consequently cell proliferation through suppression of intracellular DG level, an activator of PKC. Since the inositol 1,4,5-triphosphate level and the hydrolysis of phosphatidylcholine to DG and phosphorylcholine were not affected, forskolin therefore appears to suppress the de novo synthesis of DG.

It has been suggested that phosphorylation of myelin basic protein (MBP) in CNS is catalyzed by protein kinase C (PKC). In order to demonstrate that PKC in the myelin phosphorylates MBP, PKC was partially purified from rat CNS myelin by solubilization with Triton X-100 followed by a DEAE-cellulose column. MBP and histone III-S were phosphorylated in the presence of Ca2+ and phospholipid by rat myelin PKC. High voltage electrophoresis revealed that the phosphoamino acids in MBP by this kinase was serine residue, which is known to be the amino acid phosphorylated by PKC. The activity of PKC extracted from myelin was inhibited by the addition of psychosine to the incubation mixture. To confirm the presence of PKC molecule and to identify the isoform of PKC in the myelin, the solubilized myelin fraction was applied on SDS-PAGE, transferred to a nitrocellulose sheet and stained with anti-PKC monoclonal antibodies. Rat CNS myelin contained the PKC of about 80 kDa (intact PKC), and no proteolytic fragments were observed. PKC isozymes in myelin were type II and III. A developmental study from 14 to 42 postnatal days showed that PKC activity in CNS myelin seemed to parallel the deposition of myelin protein.

In attempts to elucidate mechanisms of demyelination in the twitcher mouse (Twi), phosphorylation and methylation of myelin basic protein (MBP) were examined in the brainstem and spinal cord of this species. Phosphorylation of MBP in isolated myelin by an endogenous kinase and an exogenous [32P]ATP was not impaired and protein kinase C activity in the brain cytosol was not reduced. When the methylation of an arginine residue of MBP was examined in slices of the brainstem and spinal cord, using [3H]methionine as a donor of the methyl groups, no difference was found between Twi and the controls. Radioactivity of the [3H] methionine residue of MBP of Twi was also similar to that of the controls. Thus, accumulation of psychosine in Twi does not interfere with the activity of endogenous kinase, methylation of MBP, and the synthesis and transport of MBP into myelin membrane.

Rolling circle amplification (RCA) generates single-stranded DNAs or RNA, and the diverse applications of this isothermal technique range from the sensitive detection of nucleic acids to analysis of single nucleotide polymorphisms. Microwave chemistry is widely applied to increase reaction rate as well as product yield and purity. The objectives of the present research were to apply microwave heating to RCA and indicate factors that contribute to the microwave selective heating effect. The microwave reaction temperature was strictly controlled using a microwave applicator optimized for enzymatic-scale reactions. Here, we showed that microwave-assisted RCA reactions catalyzed by either of the four thermostable DNA polymerases were accelerated over 4-folds compared with conventional RCA. Furthermore, the temperatures of the individual buffer components were specifically influenced by microwave heating. We concluded that microwave heating accelerated isothermal RCA of DNA because of the differential heating mechanisms of microwaves on the temperatures of reaction components, although the overall reaction temperatures were the same.

Rolling Circle Amplification (RCA) is a technique to amplify the oligoncleotides containing tandemly repetitive sequence motif. Various repetitive sequences of which the motif unit is the circular template can be synthesized by extending the primer. Additionally, RCA need not change the temperature regulatory processes like PCR, but RCA has the gene amplification efficiency similar to PCR. Although, it contains the problem of the sequence confirmation and the activity of polymerase as the DNA polymerase I. We comparatively estimated of the various polymerases for the improvement of the RCA. Five DNA polymerases were estimated for RCA.

A 69-year-old man was referred to our department because of acute onset nausea, vomiting, dysphagia, dysarthria and gait disturbance. He had a 50-day-history of amebic dysentery and had been treated with 1,500 mg metronidazole per day. Neurological examination revealed dysphagia, ataxic speech, ataxia of the left extremities and the trunk, and hyperactive deep tendon reflexes in all extremities. Sensory impairment of all modalities was apparent in a glove and stocking pattern, with mild paresthesia. Brain MRI showed T2 high signal lesions in the bilateral cerebellar dentate nuclei, more markedly on the left. On brain SPECT, obvious low blood perfusion was observed in the left cerebellar hemisphere. These findings well explained the ataxia of the left limbs. One month after discontinuing metronidazole, the cerebellar ataxia, dysphagia and MRI abnormalities completely cleared. Therefore, central nervous system damage induced by metronidazole is considered reversible. In spite of the presence of the MRI lesion in the right dentate nucleus, the patient had no ataxia of the right extremities and there was no hypoperfusion in the right cerebellar hemisphere. Thus, metronidazole does not appear to have a direct neurotoxic effect on the central nervous system. On the other hand, nerve conduction studies showed axonal polyneuropathy, which was not improved one month after cessation of the drug; thus metronidazole seems to exert more damage on peripheral nerves.

A 44-year-old man was admitted to our hospital because of a five year history of chronic progressive gait disturbance. Neurological examination revealed mild weakness and atrophy of the upper extremities, but severe of the lower ones, and without sphincter disturbance or apparent sensory impairment. Hyperreflexia and positive pathological reflexes of the lower extremities were apparent. EMG showed a reinnervation pattern and decreased number of motor units in the extremities, suggesting ALS. However, multiple plaques on the head and spinal MRI, a prolonged central conduction time of MEP and SEP, a delayed P100 latency of VEP, and a increased IgG index in the CSF indicated primary progressive type multiple sclerosis. After receiving steroid pulse therapy, the weakness of the lower extremities showed slight improvement. Diffuse inflammation in the spinal cord involving not only the pyramidal tract but also the anterior horn cells/intramedullary ventral roots explained the ALS-like clinical picture.

Acute autonomic and sensory neuropathy (AASN) is characterised by severe autonomic dysfunction, sensory deficit, and relatively well or fully preserved motor nerve function.1-4 The disease sometimes has a chronic course.2 4 Detailed information is limited because there have been so few reports of the postmortem examination of AASN.2-4 This case provides histopathological evidence for autonomic and sensory ganglionopathy in AASN. (A) Section from the lumbar dorsal root ganglia: There are only a few normal lumbar dorsal root ganglia neurons. (B) Section from the sympathetic ganglion: normal nerve cell bodies have disappeared and been replaced by clusters of cells (“nodules of Nageotte” (arrows)). Originally×100. A 30 year old Japanese man had abdominal pain and a rise in body temperature to 40°C on 1 January 1990. On 23 January, he felt prickling and paraesthesia in all his limbs. On 14 February he could no longer walk due to severe prickling of the entire body. On 2 March, he became bedridden because of orthostatic hypotension. He experienced mild weakness in the all limbs, dysarthria, and dysphagia. On 13 March, he was transferred to our hospital. Physical examination on admission showed anhidrosis, urinary retention, hyposalivation, and paralytic ileus. There was no history of intoxication by …

The Rolling Circle Amplification is an effective method of DNA amplification having tandem repeated sequences. Firstly, the template sequence is designed to form a circular structure by the annealing with the primer. The thermostable polymerase is catalyzed to primer and circular template DNA. As a result, tandem repetitive olignucleotide of circular template sequence can be obtained. In our recent study, Bst DNA polymerase catalyzed RCA reaction had been elucidated that the oligonucleotides with tandem repeated sequence were obtained as the ladder band and amplified the enough amounts of DNA within short time. On the other hand, we also focused on the microwave technology for advance to the various chemical and biological reactions. In this study, the microwave was irradiated to RCA reaction on controlling the temperature.

A Western blot analysis showed connexin32 (Cx32) to be present in the myelin membrane. Cx32 mRNA rapidly decreased in the distal segments of crushed sciatic nerves and thereafter returned to normal, as did P0 mRNA. Both Cx32 and P0 mRNAs were detectable in cultured Schwann cells and were enhanced by the addition of forskolin. The developmental profile showed that Cx32 mRNA had thus markedly increased by the time that myelination had become most active, and thereafter only slowly increased unlike P0. Cx32 mRNA seems to parallel myelin size, which thus suggests that Cx32 supports myelin maintenance as a gap junction protein that facilitates the intramyelin exchange of small molecules.

That many cells express more than one connexin (Cx) led us to examine whether Cxs other than Cx32 are expressed in the PNS. In addition to Cx32 mRNA, Cx43 and Cx26 mRNAs were detected in rat sciatic nerve by northern blot analysis. Cx43 mRNA, but not Cx26 mRNA, was expressed in both the primary Schwann cell culture and immortalized Schwann cell line (T93). The steady-state levels of the Cx43 mRNA in the primary Schwann cell culture increased 2.0-fold with 100 microM forskolin, whereas that of Po increased 7.0-fold. Immunoreactivity to Cx43 was detected on western blots of cultured Schwann cells, T93 cells, and sciatic nerves but not on blots of PNS myelin. Immunohistochemical study using human peripheral nerves revealed that anti-Cx43 antibody stained cytoplasm around nucleus of Schwann cells but not myelin, confirming western blot results. Although Po expression was markedly decreased by crush injury of the sciatic nerves, Cx43 expression showed no apparent change. Developmental profiles showed that Cx43 expression in the sciatic nerve increased rapidly after birth, peaked at about postnatal day 6, and then decreased gradually to a low level. In adult rats, the Cx43 mRNA value was much lower than that of Cx32. These findings suggest that Cx43 is localized in Schwann cell bodies and that, compared with Po, its expression is less influenced by axonal contact and cyclic AMP levels. The high expression on postnatal day 6 indicates that Cx43 may be related to PNS myelination. Cx43 is another gap junction, but its function appears to differ from that of Cx32, as judged by the differences in their localization and developmental profiles.

We report the case of a 59-year-old woman with primary Sjogren's syndrome who developed hypesthesia, hypalgesia, and neurogenic arthropathy in her lower limbs. Neurological examination and electrophysiological studies indicated involvement of the dorsal root ganglia. The immunohistochemistry of sections of rat dorsal root ganglion (DRG) showed that the IgG in the serum and cerebrospinal fluid (CSF) from the patient bound to the neuronal perikarya of small DRG neurons but not to the cerebellum or peripheral nerves. These results, consistent with particular impairment of pain and touch senses, suggest that dorsal root ganglionitis in primary Sjogren's syndrome is mediated by humoral autoimmunity.

We investigated the signal transduction system when the proliferation of cultured Schwann cells was activated by glial growth factor (GGF). When 10 micrograms/ml of GGF was added to the culture medium, Schwann cell division was activated by about 3-fold, while protein kinase C (PKC) activity was translocated from the cytosol to the particulate fraction, and diacylglycerol (DG) production was increased in the cells. PKC activity in the particulate fraction and intracellular DG content increased to 167% and 158%, respectively. However, intracellular cyclic AMP levels remained unchanged. The GGF-induced proliferation of Schwann cells was significantly inhibited by staurosporine, a PKC inhibitor, in a dose-dependent manner, while H-8, a specific inhibitor for cyclic nucleotide-dependent protein kinases, did not show an inhibitory effect even at a high concentration. These data suggest that the signal transduction system through PKC is involved in the proliferation of Schwann cells when treated with GGF.

This is the first report describing hypertrophic cranial pachymeningitis which developed in association with mixed connective tissue disease (MCTD).A 56-year-old man with a two-year history of MCTDgradually developed symptoms of headache and blurred vision. Bilateral papilledema and increased opening pressure and mild pleocytosis of the cerebrospinal fluid were noted. Magnetic resonance imaging of the brain revealed a diffuse thickening and a gadoliniumenhancement of the cranial dura mater. While five previous patients, diagnosed as idiopathic or infectious hypertrophic cranial pachymeningitis in our department, had either otitis media or sinusitis and showed multiple cranial nerve involvement, the present patient had neither otitis media nor sinusitis and did not show any cranial nerve involvement other than papilledema.(Internal Medicine 32: 510-512, 1993)