Mass producible, robust SERS substrates based on metal film on nanosphere (MFON) on an adhesive substrate for detection of surface-adsorbed molecules and their evaluation by helium ion microscopy.

We have developed a SERS stamp that can be pressed directly onto a solid surface for characterization of surface-adsorbed target molecules. The stamp was fabricated by transfer of a dense monolayer of SiO2 nanospheres from a glass surface onto a piece of adhesive tape and subsequent evaporation of silver. The performance of the resulting SERS stamps was evaluated by their exposure to methyl mercaptan vapor, and immersion in rhodamine 6G and ferbam solutions. It was found that beside the nanosphere diameter and metal deposition thickness, the extent of burial of the nanospheres into the adhesive tape, dictated by the pressure during the nanosphere transfer process, had a significant effect. We carried out FDTD calculations of the near field. Models are based on morphological information obtained from helium ion microscopy, which can provide high-resolution images of poor electrical conductors such as our SERS stamp. While one of our main eventual goals is detection of pesticides on agricultural produce, we have begun to take a careful step by testing our SERS stamp on better characterized surfaces such as a porous gel surface, having been immersed in fungicides such as ferbam. We also present our preliminary results with ferbam on oranges. It is expected that our well-characterized SERS stamp will play a role in shedding light on the poorly studied transfer process of target molecules onto a SERS surface as well as serving as a new SERS platform.

Listening evaluation of cochlear implant users: comparison of subjective and objective evaluation by visual analogue scale.

OBJECTIVE: This study aimed to use short-form visual analogue scale cochlear implantation questionnaires to evaluate subjective aspects at each out-patient visit. The correlation between subjective hearing tests using the short-form visual analogue scale and objective hearing outcomes was evaluated. METHOD: This study was conducted in a single centre. Cochlear implant users (n = 199) evaluated their hearing on a scale of 0 to 100 for the right, left and both ears. The Japanese speech perception test (CI-2004) Japanese monosyllable speech perception test (67-S) and cochlear implantation threshold were used for the objective cochlear implantation evaluation. RESULTS: A significant correlation was found between the short-form visual analogue scale questionnaire and objective hearing outcome, for words (r = 0.64) and sentences (r = 0.62) in CI-2004 and 67-S (r = 0.56) tests. No significant correlation was found between the short-form visual analogue scale score and cochlear implantation threshold (r = -0.18). CONCLUSION: Short-form visual analogue scale cochlear implantation questionnaires mean cochlear implant users spend less time answering subjective visual analogue scale questionnaires, and clinicians estimate a patient's cochlear implantation hearing and abnormality by chronological evaluation.

Differential distribution of vesicular glutamate transporters in the rat cerebellar cortex.

The chemical organization of excitatory axon terminals in the rat cerebellar cortex was examined by immunocytochemistry and in situ hybridization histochemistry of vesicular glutamate transporters 1 and 2 (VGluT1 and VGluT2). Chemical depletion of the inferior olivary complex neurons by 3-acetylpyridine treatment almost completely removed VGluT2 immunoreactivity from the molecular layer, leaving VGluT1 immunoreactivity apparently intact. On the other hand, neuronal deprivation of the cerebellar cortex by kainic acid injection induced a large loss of VGluT1 immunoreactivity in the molecular layer. In the cerebellar granular layer, both VGluT1 and VGluT2 immunoreactivities were found in mossy fiber terminals, and the two immunoreactivities were mostly colocalized in single-axon terminals. Signals for mRNA encoding VGluT2 were found in the inferior olivary complex, and those for VGluT1 and VGluT2 mRNAs were observed in most brainstem precerebellar nuclei sending mossy fibers, such as the pontine, pontine tegmental reticular, lateral reticular and external cuneate nuclei. These results indicate that climbing and parallel fibers selectively use VGluT2 and VGluT1, respectively, whereas mossy fibers apply both VGluT1 and VGluT2 together to accumulate glutamate into synaptic vesicles. Since climbing-fiber and parallel-fiber terminals are known to make depressing and facilitating synapses, respectively, VGluT1 and VGluT2 might have distinct properties associated with those synaptic characteristics. Thus, it would be the next interesting issue to determine whether mossy-fiber terminals co-expressing VGluT1 and VGluT2 show synaptic facilitation or depression.

In vivo transduction of central neurons using recombinant Sindbis virus: Golgi-like labeling of dendrites and axons with membrane-targeted fluorescent proteins.

A new recombinant virus which labeled the infected neurons in a Golgi stain-like fashion was developed. The virus was based on a replication-defective Sindbis virus and was designed to express green fluorescent protein with a palmitoylation signal (palGFP). When the virus was injected into the ventrobasal thalamic nuclei, many neurons were visualized with the fluorescence of palGFP in the injection site. The labeling was enhanced by immunocytochemical staining with an antibody to green fluorescent protein to show the entire configuration of the dendrites. Thalamocortical axons of the infected neurons were also intensely immunostained in the somatosensory cortex. In contrast to palGFP, when DsRed with the same palmitoylation signal (palDsRed) was introduced into neurons with the Sindbis virus, palDsRed neither visualized the infected neurons in a Golgi stain-like manner nor stained projecting axons in the cerebral cortex. The palDsRed appeared to be aggregated or accumulated in some organelles in the infected neurons. Anterograde labeling with palGFP Sindbis virus was very intense, not only in thalamocortical neurons but also in callosal, striatonigral, and nigrostriatal neurons. Occasionally there were retrogradely labeled neurons that showed Golgi stain-like images. These results indicate that palGFP Sindbis virus can be used as an excellent anterograde tracer in the central nervous system.

[Relationship between clinical activity and lymphocyte subsets in patients with Guillain-Barré syndrome].

T lymphocytes are probably of pathogenic importance in Guillain-Barré syndrome (GBS). Blood samples from 31 patients with GBS and from 25 healthy controls were studied with flow cytometry. Subsets of the CD4 population were determined: CD4+CD29+ (helper-inducer) cells and CD4+CD45RA+ (naive or suppressor-inducer) cells. Recently, deviations of circulating CD8 populations have been noticed. Therefore, subsets of the CD8 population were also determined: CD8+CD11b dull (suppressor-effector) cells and CD8+ CD11b bright (cytotoxic-effector/NK) cells. During the active stage of GBS, significantly increased proportions of CD4+HLA-DR+ (activated CD4+) cells and CD4+CD29+ cells were found compared to those in control samples (p < 0.0005, p < 0.05, respectively). The proportion of CD8+CD11b dull cells was significantly lower than that in the control samples (p < 0.005). Sequential analysis showed a decrease in the proportion of CD4+HLA-DR+ cells and CD4+CD29+ cells from the active to the convalescent stage (p < 0.05, p < 0.005, respectively). The proportion of CD8+CD11b dull cells was low in GBS patients with high disease severity.

[Two cases of encephalo-myelo-radiculoneuropathy, triggered by herpes simplex virus type-1 infection].

We report two cases of encephalo-myelo-radiculoneuropathy, triggered by herpes simplex virus type-1 (HSV-1) infection. Patient 1 (a 25-year-old man) and patient 2 (a 52-year-old man) were admitted to the hospital because of fever, headache, abnormal behavior, and loss of consciousness. In each case, cerebrospinal fluid (CSF) showed lymphocytic pleocytosis with protein elevation, and serum and CSF IgG antibody titers to HSV-1 were elevated markedly. Although patient 1 was treated with aciclovir in the early phase of encephalitis, he developed severe quadriparesis as a sequela. Patient 2 was treated with a combination of aciclovir and corticosteroids, and he recovered completely about 4 months after the onset of the disease. There have been only a few reports of encephalo-myelo-radiculoneuropathy triggered by HSV-1 infection. Early corticosteroid therapy was effective in our patients with post-HSV-1 infectious encephalo-myelo-radiculoneuropathy. These two patients were studied with flow cytometry for peripheral blood lymphocyte subsets during the disease course. In the active stage of the disease, the helper-inducer (CD4 + CD29+), activated T cell (CD4 + CD25+), and cytotoxic/NK (CD8 Dull + CD11b Bright+) subsets were increased compared with subsets in controls. An interesting finding was mismatched responses with an increased suppressor-inducer (CD4 + Leu8+) subset and a decreased suppressor-effecter (CD8 Bright+ CD11b Dull+) subset, indicating a possible autoimmune character of encephalo-myelo-radiculoneuropathy triggered by viral infection.

[A case of spinal segmental myoclonus and propriospinal myoclonus: a neuroelectrophysiologic analysis].

A 37-year-old male patient with spinal segmental myoclonus and propriospinal myoclonus was described. He was admitted to our hospital because of paroxysmal axial myoclonus, which first appeared one month before. He denied any significant accident such as trauma or fever. Apart from myoclonus, no abnormal findings were observed by physical and neurological examinations, routine laboratory investigations and MRI of the cervical and thoracic spinal cords. The myoclonus consisted of continuous rhythmic contractions of the bilateral thoracal and abdominal muscles. Its frequency was approximately 0.3Hz. The clinical findings were typical of spinal segmental myoclonus. In addition, the myoclonus started in the thoracal muscles and frequently spread up to the neck muscles and down to the leg muscles. The myoclonus disappeared in sleep. Polymyography revealed the following findings: (1) The jerks were found on the bilateral axial muscles including sternocleidomastoid, biceps, triceps, pectoralis major, abdominal muscles and quadriceps. (2) Homologous muscles were activated synchronously. (3) The duration of bursts was variable ranging 100 to 400msec. (4) The jerks in the pectoralis muscle preceded those in other muscles. The latencies of the jerks in the other muscles increased with their distance from pectoralis, based on the linear regression analysis of the onset of jerks in the various muscles. (5) The jerks were induced by tapping or electrical stimulation anywhere on the body including the face, but not by flash or sound. From the above polymyographical findings, the myoclonus seems to originate in the T3 spinal cord and slowly up and down the spinal cord at 0.6-1.6 m/sec, suggesting that it is mediated by the propriospinal tract.

[Polymyalgia rheumatica and myelitis associated with anti-cardiolipin antibody].

A 78-year-old woman was admitted to our hospital on September 14, 1992, because of systemic myalgia and stiffness, joint pain, and gait disturbance. She had begun to feel headache and pain in the neck and shoulder in the middle of August, 1992. The pain became systemic, and was accompanied by a low-grade fever, which was unresponsive to NSAIDs. On admission, she had no joint swelling or deformities in the extremities. Neurological examination revealed weakness in the right leg, hypoalgesia below the left C4 level, hyperreflexia in the right extremities, and right Babinski's sign. The erythrocyte sedimentation rate was very high (100 mm/h). Levels of other acute phase reactants were also high. Tests for antinuclear antibody and anti-cardiolipin antibody were positive, but a test for rheumatoid factor was negative. Creatine kinase activity was within normal limits. A T1-weighted magnetic resonance image of the cervical spine at 0.5 T showed an intramedullary low signal. A T2-weighted image showed a borderless spindle-like high signal. Four nodules enhanced by Gd DTPA were seen at C1-C4. The age at onset, myalgia, stiffness, and erythrocyte sedimentation rate were considered to be consistent with a diagnosis of polymyalgia rheumatica. Glucocorticoid treatment was therefore started, and a dramatic clinical improvement was evident within a few days. The patient was discharged from hospital on November 30, 1992. To our knowledge, myelopathy complicated by polymyalgia rheumatica has never been reported previously. Recently, some patients with polymyalgia rheumatica have been reported to have anti-cardiolipin antibody in serum. In the present case anti-cardiolipin antibody may have played a role in the formation of microemboli or in angitis of the cervical spine.

Pathological examination with two-millimetre serial sectioning for sentinel lymph node biopsies in breast cancer.

AIMS: To compare enhanced pathology with serial sectioning and the transcription-reverse transcription concerted reaction (TRC) for detecting sentinel node (SN) metastasis in breast cancer cases. METHODS: In total, 115 SN samples from 32 breast cancer cases were investigated by pathological examination with 2.0-mm serial sectioning and by quantitative analysis of carcinoembryonic antigen messenger RNA with the TRC. RESULTS: The results were concordant in 98.3% of these cases. Two histologically metastatic nodes tested negative by TRC, whereas none tested positive by TRC alone. CONCLUSION: Pathological examination with 2-mm sectioning showed superior performance to TRC under the study conditions.

Gold-induced pulmonary disease: clinical features, outcome, and differentiation from rheumatoid lung disease.

Gold-induced pulmonary disease is difficult to diagnose, especially, in the case in which interstitial pneumonia appears in the course of gold therapy for rheumatoid arthritis. We analyzed the literature to define the clinical features and prognosis of gold-induced pulmonary disease, and to identify those features that distinguish gold-induced pulmonary disease from pulmonary disease secondary to the underlying disease process of rheumatoid arthritis. Relevant articles from the medical literature were identified using a Mediline search, and the bibliographies of the articles were retrieved. These works were critically reviewed for information on the clinical, physiologic, radiographic, pathologic, and bronchoalveolar lavage (BAL) findings. A total of 140 cases of gold-induced pulmonary disease were identified from 110 reports. In 81% of the patients, gold was being used to treat rheumatoid arthritis, bronchial asthma (6%), pemphigus (5%), or other processes (9%). Side effects other than pulmonary toxicity were common, and included skin rash (38%), peripheral eosinophilia (38%), liver dysfunction (15%), and proteinuria (22%). Only the presence of pemphigus or liver dysfunction correlated with a bad prognosis. Gold-induced pulmonary disease most often followed improvement in rheumatoid arthritis, presumably induced by gold therapy. BAL lymphocytosis and computed tomography (CT) scan findings are useful in making a diagnosis of gold-induced pulmonary disease in an appropriate clinical setting. Features that distinguish gold-induced pulmonary disease from rheumatoid lung disease include female predominance, presence of fever or skin rash, absence of subcutaneous nodules or finger clubbing, low titers of rheumatoid factor at onset of lung disease, lymphocytosis in bronchoalveolar lavage fluid (BALF), and alveolar opacities along the bronchovascular bundles on chest CT scan. Gold-induced lung disease is a distinct entity that can be distinguished from rheumatoid lung disease. It usually improves with cessation of therapy or treatment with corticosteriods.

An alanine racemase gene as a new index for detecting Escherichia coli in foods.

A gene of alanine racemase, a typical prokaryotic enzyme, was evaluated as a new index for detecting Escherichia coli in foods. An alanine racemase gene fragment containing a non-conserved sequence of the gene was amplified from genomic DNA of E. coli by a polymerase chain reaction, and then labeled with digoxigenin as a probe for detecting E. coli. Food samples and bacteria were each treated as at 25 degrees C for 10 min in 0.1N NaOH containing 0.5% SDS, before being directly spotted on to nylon membranes for DNA hybridization. The probe was specific for E. coli; all 48 strains of E. coli examined, including such pathogenic strains as E. coli O157:H7, showed positive signals, whereas all 59 strains of non-E. coli species, except for one strain (Shigella sonnei), did not show a signal. Various foods inoculated with E. coli K-12 showed positive signals whereas no uninoculated foods showed any signal. Quantification of E. coli cells in the death phase by the hybridization method showed good correlation with that by the plate culture method. The alanine racemase gene could prove useful as an index for detecting E. coli in foods.

Comparative study of efficacy of plasma exchange versus intravenous gammaglobulin treatment on acute postinfectious polyradiculoneuropathy: a preliminary report.

We tried to compare the efficacy of plasma exchange (PE) with that of intravenous immunoglobulin (IVIG) in patients with postinfectious polyneuritis (Guillain-Barré syndrome [GBS] and cranial neuritis). Fifteen patients with postinfectious polyneuritis were divided into 2 groups. The IVIG group included 5 cases of GBS and 2 cases of postinfectious cranial neuritis (ophthalmoplegic type). The PE group included 5 cases of GBS and 3 cases of postinfectious cranial neuritis (ophthalmoplegic type). The changes and incidences of improvement of muscle strength scores (MSSs) and ocular movement scores (OMSs) were evaluated before treatment and 1, 2, 3, and 4 weeks after treatment. No significant differences between the IVIG and PE groups were found in the MSSs or OMSs at any time after treatment. These data suggested that PE and IVIG had equivalent efficacy. In the IVIG group, the proportion of suppressor-inducer T cells significantly increased (p < 0.01) (before versus after treatment), and the proportion of suppressor-effector cells also increased but not significantly (before versus after treatment). In the PE group, the percentage of suppressor-inducer T cells significantly decreased (p < 0.05) (before versus after treatment) while the proportion of suppressor/cytotoxic T cells significantly increased (p < 0.05) (before versus after treatment). The percentage of suppressor-effector T cells also increased (before versus after treatment) but not significantly.