Development and evaluation of a loop-mediated isothermal amplification method for the rapid detection of Chlamydophila pneumoniae.

We developed a loop-mediated isothermal amplification (LAMP) method to detect Chlamydophila pneumoniae infection. This assay exclusively amplified C. pneumoniae sequences and no cross-reactivity was observed for other Chlamydia species. The detection limit for this assay was found to be ten elementary bodies in 25 min, as observed in a real-time turbidimeter and electrophoretic analysis. The specificity of the LAMP reaction was confirmed by restriction endonuclease analysis, as well as direct sequencing of the amplified product. Among nasopharyngeal swab specimens from 120 patients with acute respiratory tract infections and 40 healthy individuals, the LAMP results showed 100% agreement with the results of real-time polymerase chain reaction (PCR) assays.

[Off-pump coronary artery bypass surgery for the ischemic cardiomyopathy patient with moderate mitral regurgitation and stenosis of bilateral vertebral arteries].

We report a 71-year-old man who had severe coronary and cerebral vascular disease with moderate mitral regurgitation (MR). Left ventricular reconstruction and mitral valve surgery were considered for poor left ventricular function and dilatation with MR. However, low blood pressure during cardiac arrest was risk for stroke due to severe stenosis of bilateral vertebral arteries. The myocardial viability of the anterior wall and inferior wall was confirmed by thallium-201 rest-redistribution single photon emission computed tomography (SPECT). Therefore, off-pump coronary artery bypass grafting (OPCAB) was selected for the patient. Only three Lima sutures were used for keeping the optimal heart position. Coronary anastomoses were done in sequence right gastroepiploic artery (RGEA)-#4 posterior descending (PD), left internal thoracic artery (LITA)-#9-#14 (sequential), RITA-#8 left anterior descending (LAD). No neurological complication occurred postoperatively. Left ventricular function and MR gradually improved. Final ejection fraction (EF) is 51% and MR is trivial. This case demonstrated improvement of MR by only revascularization according to preoperative viability assessment.

Contaminant-associated disruption of vitamin A and its receptor (retinoic acid receptor alpha) in free-ranging harbour seals (Phoca vitulina).

Polychlorinated biphenyls (PCBs) have been associated with a number of toxic effects in marine mammals such as endocrine disruption and immunotoxicity that, in turn, are widely thought to have contributed to population level impacts including reproductive failure and outbreaks of disease. In this study, the dietary hormone vitamin A and expression levels of one of its receptors, retinoic acid receptor alpha (RARalpha), were used as biomarkers of PCB-associated health effects in harbour seals. Harbour seal pups (n=24) were live-captured in coastal British Columbia, Canada, and Washington State, USA, and sampled for whole blood (to obtain peripheral blood mononuclear cells, PBMCs) and blood plasma, as well as biopsies of blubber and skin. Concentrations of circulatory vitamin A (retinol) in plasma and stored vitamin A in blubber were negatively associated with blubber PCB concentrations (R=-0.518, p=0.013 and R=-0.645, p=0.009, respectively). However, vitamin A concentrations in skin, an important target tissue, remained constant, which likely reflects a compensatory transfer from blubber to maintain physiological functions. In addition, we characterized the harbour seal RARalpha, and investigated its expression levels as a potential biomarker in seals. RARalpha expression in blubber, but not on PBMCs, was elevated in more contaminated animals (R=0.580, p=0.009). This may represent a direct contaminant-related effect, or, a compensation for the contaminant-related disruption of (circulatory and/or blubber) hormone levels. Since vitamin A is critical to developmental, reproductive and immunological health, our observations of a contaminant-related disruption of its physiology in free-ranging seals may portend population level consequences. Vitamin A concentrations and RARalpha expression levels can therefore represent relevant and sensitive biomarkers of PCB-associated toxic effects in toxicological studies of marine mammals.

Comparison of whole genome sequences of Chlamydia pneumoniae J138 from Japan and CWL029 from USA.

Chlamydia pneumoniae is a widespread pathogen of humans causing pneumonia and bronchitis. There are many reports of an association between C.PNEUMONIAE: infection and atherosclerosis. We determined the whole genome sequence of C.PNEUMONIAE: strain J138 isolated in Japan in 1994 and compared it with the sequence of strain CWL029 isolated in the USA before 1987. The J138 circular chromosome consists of 1 226 565 nt (40.7% G+C) with 1072 likely protein-coding genes that is 3665 nt shorter than the CWL029 genome. Plasmids, phage- or transposon-like sequences were not identified. The overall genomic organization, gene order and predicted proteomes of the two strains are very similar, suggesting a high level of structural and functional conservation between the two unrelated isolates. The most conspicuous differences in the J138 genome relative to the CWL029 genome are the absence of five DNA segments, ranging in size from 89 to 1649 nt, and the presence of three DNA segments, ranging from 27 to 84 nt. The complex organization of these 'different zones' may be attributable to a unique system of recombination.

Complete nucleotide sequence of human NRAMP2 cDNA.

The mouse Lsh/Ity/Bcg locus regulates macrophage activation for antimicrobial activity against intracellular pathogens, and the Nramp1 gene was isolated as its candidate. In the present study, a full length cDNA for human NRAMP2 has been isolated and characterized. Nucleotide sequence analysis reveals that the cDNA, 4142 bp in length, coded for a protein of 561 amino acid residues with a molecular weight mass of 61,456. Predicted amino acid sequence analysis of the NRAMP2 molecule indicates that NRAMP1 and NRAMP2 sequences share 64% identical residues overall, whereas only 21% at the NH2-terminal cytoplasmic domain, where the NRAMP1 molecule was associated with microtubules, was found to be identical. This suggests that the NH2-terminal region of the NRAMP2 molecule may have a particular function, different from that of the NRAMP1 molecule.

Ataxic hemiparesis from small capsular hemorrhage. Computed tomography and somatosensory evoked potentials.

We studied three cases that fit the clinical syndrome of ataxic hemiparesis. Computed tomography revealed small hemorrhages of the posterior limb of the internal capsule in all the cases, and somatosensory evoked potential studies predicted a disturbance of the lemniscal pathway from the thalamus to the parietal sensory cortex without demonstrable sensory loss. These results suggested that ataxia of the contralateral limbs following capsular lesion might result from a disruption of the thalamocortical sensory projection. The site of the lesion, the pathologic cause, and the underlying mechanism of ataxia are not necessarily homogeneous within the syndrome.

Spontaneous dural carotid cavernous sinus fistula presenting isolated ophthalmoplegia: evaluation with MR angiography.

We describe three patients with spontaneous dural carotid cavernous sinus fistula presenting an isolated ophthalmoplegia where magnetic resonance angiography demonstrated abnormal flow-related enhancements in the cavernous sinus with extension into the inferior petrosal sinus. Magnetic resonance angiography is of value in evaluation of patients with isolated ophthalmoplegia.

Intravenous recombinant tissue plasminogen activator in acute carotid artery territory stroke.

To determine the effect of intravenous recombinant tissue plasminogen activator (rt-PA) on vascular and neurologic outcomes, we enrolled 31 patients with acute carotid artery-territory ischemic stroke within 6 hours from symptom onset in a randomized, double-blind, placebo-controlled study. We gave either rt-PA (duteplase at the dose of 20 or 30 mega-international units [MIU]) or placebo intravenously for 60 minutes in patients randomly assigned to the three groups. A comparison between the baseline and postinfusion angiograms showed that complete or partial reperfusion occurred in 50% (5/10) of patients treated with 30 MIU rt-PA, 44% (4/9) of those treated with 20 MIU rt-PA, and 17% (2/12) in the control group. In patients with middle cerebral artery occlusions, reperfusion occurred in 71% (5/7) of the 30-MIU group, in 67% (4/6) of the 20-MIU group, and in 13% (1/8) of the control group. Patients treated with 30 MIU rt-PA showed a significantly early and better clinical improvement, as measured by the neurologic scale, than did those treated with placebo. Parenchymal hemorrhage occurred in one patient in each group, and frequency of clinically insignificant hemorrhagic infarction was comparable among the treatment groups. No major systemic complications occurred in any group. These results support the efficacy of intravenous infusion of rt-PA soon after the onset of stroke in producing rapid thrombolysis and neurologic recovery; it may be of particular value in patients with thromboembolic occlusion in the middle cerebral artery.

Pure anarthria with predominantly sequencing errors in phoneme articulation: a case report.

A 77-year-old left-handed man presented with pure anarthria following cerebral infarction. The lesion was restricted to the right precentral gyrus extending to the immediately underlying subcortical white matter and the frontal part of the insular cortex. Qualitative analysis of anarthria revealed that half of the phonemic-articulatory errors in spontaneous speech were sequencing ones. Sequential errors were detected at the phoneme level in both consonants and vowels, and at the syllable level. Most of the sequential errors were pre-positioning. Sequential errors were observed both within and across words. In clear contrast with anarthria, writing and comprehension was preserved, which suggested the problem was limited to oral expression. Our findings provide further support that the precentral gyrus and/or the insular cortex of the language dominant hemisphere is responsible for the temporal sequencing of the articulatory programming.

The effect of CV-3988 and CV-6209 on the acute gastric erosions of rats due to water-immersion and restraint stress.

CV-3988 and CV-6209 inhibited gastric erosions in rats due to water-immersion and restraint stress in a dose-dependent manner. The above inhibitory effects of CV-3988 were observed in the presence of indomethacin, which may indicate that the inhibition is not prostaglandin dependent. The studies indicate that platelet-activating factor may be involved in the formation of erosions in rats under water-immersion and restraint stress.

Crossed non-dominant hemisphere syndrome in a right-hander.

A right-handed patient with a large left temporo-parietal infarction manifested various non-dominant hemisphere signs. He had two left-handed children. On neurobehavioural examinations, he did not show aphasia or ideomotor apraxia, but did show hemispatial neglect, spatial agraphia, constructional apraxia, auditory and tactile extinction, anosodiaphoria and affective changes, all of which are usually observed after right hemispheric damage. We conclude that he has a reversed cerebral laterality of cognitive functions and showed crossed non-dominant hemisphere syndrome.

Hyperlalia: a right cerebral hemisphere syndrome.

We describe a new right hemispheric behavioural syndrome for which we propose the name "hyperlalia". In a typical case an apparently unconcerned and expressionless patient is easily prompted to remarkable volubility with a content which is loose and incoherent. The voice is low and monotonous. All the lesions confirmed by computed tomography of the brain overlapped in the perisylvian area in the territory of the right middle cerebral artery.Loss of a subtle balance between the left hemispheric speech area and the corresponding area in the right hemisphere caused by acute damage of the right perisylvian area may have resulted in disinhibition of the speech function. Similarities and dissimilarities with the known pathology of talkativeness are also discussed.

A new mouse model of spontaneous diabetes derived from ddY strain.

By the selective breeding of obese male mice of the ddY strain and using indices of the heavy body weight and appearance of urinary glucose, we established two inbred strains in 1992: one with obesity and urinary glucose (Tsumura, Suzuki, Obese Diabetes: TSOD) and the other without them (Tsumura, Suzuki, Non Obesity: TSNO). The male TSOD mice constantly showed signs of obesity and urinary glucose with increases in food and water intake, body weight and some fat weight. The body mass index (BMI) clearly showed moderate obesity. Increases in the levels of diabetic blood parameters (glucose, insulin and lipids) were also found in males, in which the levels of blood glucose and insulin were high to the ages past the growth peak. In the histological studies, pancreatic islets of the TSOD males were found hypertrophic without any signs of insulitis or fibrous formation. Among these diabetic characteristics, some of which were similar to the reported models of non-insulin-dependent diabetes mellitus (NIDDM), the stable appearances of the hyperglycemia, the hyperinsulinemia and the hypertrophy of pancreatic islets to the ages past the growth peak were the prominent features. In these respect the TSOD mouse may be a useful model for researching the mechanisms of human diabetes and its complications.

Fluorescent derivatization of nitrite ions with 2,3-diaminonaphthalene utilizing a pH gradient in a Y-shaped microchannel.

The on-chip derivatization of nitrite ions with 2,3-diaminonaphthalene (DAN) utilizing a pH gradient formed in a Y-shaped microchannel was investigated. Nitrite ions react with DAN at low pH, and strongly fluoresced at high pH. Therefore, a reaction at low pH followed by the addition of a strong alkaline solution is the usual procedure in a batch scheme. However, a strong alkaline solution, like an NaOH aqueous solution, erodes the wall of the microchannels in substrates made of glass or polymers, and has not been considered suitable for use in microchannels. We first investigated the derivatization reaction and fluorescent properties of nitrite ions with DAN. We found that the on-chip fluorescent derivatization reaction and detection without the addition of an alkaline solution is possible by controlling the pH values of the nitrite solution and the DAN solution to form a suitable pH gradient by utilizing a buffering effect of triethanolamine solution, which is used as an NO2 gas-absorption medium. These results have suggested the feasibility of novel reaction schemes which can provide the desired products due to a controlled pH gradient in the microchannels, as well as the possibility of an on-site monitoring microchip device for ambient NO2.

[A case of moyamoya-like disease with repetitive TIA on drinking].

We reported here a patient of moyamoya-like disease with repetitive limb-shaking TIAs on drinking. Regional cerebral blood flow (rCBF) was quantified with and without drinking by single photon emission computed tomography with intravenously injected N-isopropyl-p-(I-123) iodoamphetamine (IMP). RCBF was reduced by alcohol ingestion. We speculated that homodynamic TIAs might be caused by impaired autoregulation in the region of the left carotid artery.

[The role of magnetic resonance imaging in the diagnosis of Hallervorden-Spatz disease].

A mentally-retarded 21-year-old female with narrowing of the visual field was admitted to our hospital because of dystonia in the body and extremities which appeared 6 months before and was gradually exacerbating. On admission, torsion dystonia, pyramidal tract sign, and retinitis pigmentosa with optic atrophy, urinary incontinence, hyperhidrosis and insomnia were noted. On the Wechsler Adult Intelligence Scale, she achieved less than 60 for verbal-IQ. Laboratory tests including serum copper and ceruloplasmin, were all normal. A computed tomography of the brain showed small hyperdense spot in the globus pallidus on the both sides. There was neither cortical atropy nor ventricular dilatation. A Magnetic Resonance Imaging was obtained using both moderate-field (0.5 Tesla) and high-field (1.5 Tesla) superconducting MR systems. T2-weighted spin echo images demonstrated markedly decreased signal intensity area restricted in the globus pallidus and substantia nigra on both system. Moreover, in the center of the globus pallidus, a small high signal spot was seen ("eye-of the-tiger" sign coined by Sethi et al). There was no abnormal signal in the putamen, caudate nucleus, red nucleus, and dentate nucleus. On the other hand, T1-weighted inversion recovery images were not remarkable. These striking low attenuation on T2-weighted findings suggests an increased iron deposition limited to the globus pallidus and substantia nigra which is characteristic of Hallervorden-Spatz disease/syndrome (H-S). Moreover, "eye-of-the-tiger" sign on T2-weighted MR images in the globus pallidus is previously described in H-S.(ABSTRACT TRUNCATED AT 250 WORDS)

[Occlusive lesions of carotid and intracranial arteries in patients with symptomatic lacunar infarction--evaluation by MR angiography].

The purpose of this study is to elucidate the prevalence and degree of occlusive lesions of carotid and intracranial arteries in patients with symptomatic lacunar infarction. We performed carotid and intracranial MR angiography, and T2-weighted MRI on 65 patients with symptomatic lacunar infarction. Stenosis of more than 25% narrowing of the diameter was found in 12 patients (18.5%) in the extracranial carotid arteries and in 14 patients (21.5%) in the intracranial arteries. Most of the stenotic lesions were mild. Multiple logistic regression analysis showed that age was significant and independent predictor for carotid artery stenosis, and that diabetes mellitus was predictor for intracranial artery stenosis. The incidence of intracranial artery stenosis was statistically higher in patients with asymptomatic lacunar infarctions in subcortical white matter area on T2-weighted MRI than in patients without such lesions. Coexistence of carotid artery stenosis should be suspected in aged patients with symptomatic lacunar infarction, and the possibility of intracranial artery stenosis should be considered in patients with diabetes mellitus or asymptomatic lacunar infarctions in subcortical white matter area.

[Warfarin therapy and chlorella].

Warfarin potassium (warfarin) is one of anticoagulants. Its anticoagulant effect is induced by inhibiting vitamin K in a complex manner. It is used effectively and safely in preventing cerebral embolism of cardiac origin, provided that thrombotest (TT) values is maintained from 10 to 20%. A 75-year-old man with atrial fibrillation and cardiomyopathy had cerebral embolism in the territory of the right middle cerebral artery. Warfarin therapy was started to prevent the recurrence of embolic stroke. TT values had been well controlled until intake of chlorella, but they rose above the therapeutic limit after its intake. There was no evidence of discontinuing warfarin, or taking drugs containing vitamin K or Natto. Chlorella is one of vitamin K-rich foods. Thus, it may inhibit the anticoagulant effect of warfarin.

[Trigeminal motor paralysis and dislocation of the temporo-mandibular joints].

A 64-year-old woman with diabetes mellitus was admitted to our hospital with left hemiparesis of sudden onset. A brain MRI demonstrated a cerebral infarction in the ventral part of the right lower pons. When left hemiparesis worsened, she had dislocation of the temporo-mandibular joints repeatedly. Then, her lower jaw deviated to the right when she opened her mouth. Also, there was decreased contraction of the right masseter when she clenched her teeth. These findings suggest that there was trigeminal motor paralysis on the right side resulting from involvement of the intrapontine trigeminal motor nerve. She has no history of dislocation of the temporo-mandibular joints. An X-ray film showed that the temporo-mandibular joints were intact. Thus, it is possible that deviation of the lower jaw was the cause of this dislocation. We suspect that dislocation of the temporo-mandibular joints may occur as a complication of unilateral trigeminal motor paralysis. This has not been reported to our knowledge.

[Sequential magnetic resonance imaging and neurophysiological studies in a patient with inferior spinal cord infarction].

A 66-year-old woman suddenly developed anterior spinal artery syndrome with complete flaccid paraplegia, superficial sensory disturbance caudally to the L5 dermatome level with preservation of deep sensation, incontinence, and absent deep tendon reflexes in both legs. An MRI of the whole spine and an analysis of the CSF 4 hours after onset were normal. The electrophysiological study showed an absence of F wave on the posterior tibial nerve stimulation on admission, while the peripheral nerve conduction velocities and amplitudes of upper and lower limbs were normal. T2-weighted MRI 4 days after onset demonstrated an area of high signal intensity in the gray and white matters of the epiconus and conus medullaris, and T1-weighted MRI showed the swelling. Three weeks later, F wave became evoked nd the high signal areas on axial T2-weighted MRI were localized in the bilateral anterior horns of the gray matter. T1-weighted MRIs after an administration of Gd-DTPA 3 and 7 weeks after onset demonstrated an enhancement of the ventral roots of the lumbar nerves and cauda equina, while the enhancements disappeared 8 months after onset. The patient was finally able to walk independently over 10 meter. An absent F wave was an only positive finding at the hyperacute (hours after onset) stage of the spinal cord infarction.