Applications of magnetic and electromagnetic forces in micro-analytical systems.

Novel applications of magnetic fields in analytical chemistry have become a remarkable trend in the last two decades. Various magnetic forces have been employed for the migration, orientation, manipulation, and trapping of microparticles, and new analytical platforms for separating and detecting molecules have been proposed. Magnetic materials such as functional magnetic nanoparticles, magnetic nanocomposites, and specially designed magnetic solids and liquids have also been developed for analytical purposes. Numerous attractive applications of magnetic and electromagnetic forces on magnetic and non-magnetic materials have been studied, but fundamental studies to understand the working principles of magnetic forces have been challenging. These studies will form a new field of magneto-analytical science, which should be developed as an interdisciplinary field. In this review, essential pioneering works and recent attractive developments are presented.

Association study of TRAF1-C5 polymorphisms with susceptibility to rheumatoid arthritis and systemic lupus erythematosus in Japanese.

OBJECTIVE: The primary aim of this study was to investigate the association of polymorphisms of TRAF1-C5, a newly identified rheumatoid arthritis (RA) risk locus in Caucasians, with susceptibility to RA and systemic lupus erythematosus (SLE) in Japanese populations. Gene expression levels of TRAF1 and C5 to assess the functional significance of genotypes were also analysed. METHODS: A multicentre association study consisting of 4 RA case-control series (4397 cases and 2857 controls) and 3 SLE case-control series (591 cases and 2199 shared controls) was conducted. Genotyping was performed using TaqMan genotyping assay for two single nucleotide polymorphisms (SNPs) that showed the best evidence of association in the previous Caucasian studies. Quantifications of TRAF1 and C5 expression were performed with TaqMan expression assay. RESULTS: Significant differences in allele frequency for both SNPs were observed between RA and control subjects (combined odds ratio = 1.09), while no significant difference was detected between patients with SLE and controls. Interestingly, alleles rs3761847 A and rs10818488 G had increased the risk for RA in the present study, while they decreased the risk in the original studies. A significant difference was found between risk allele carriers and non-carriers of rs10818488 for the expression level of TRAF1 in phorbol myristate acetate-stimulated lymphoblastoid cell lines (p = 0.04). CONCLUSION: Association of TRAF1-C5 locus with RA susceptibility was detected in the Japanese populations with modest magnitude, while no significant association was observed for SLE. Significant positive effect of genotype on the expression of TRAF1 might support the genetic association between TRAF1 and RA.

Selective expression of MHC class I in the affected muscle of a patient with idiopathic inflammatory myopathy.

The dominant clinical feature of polymyositis/dermatomyositis is weakness in proximal, rather than distal, musculature. Although rare, cases of focal/localized myositis in which polymyositis-like muscle inflammation is present in only one muscle or extremity have also been reported. The underlying mechanisms dictating involvement of specific muscle groups in polymyositis/dermatomyositis and focal/localized myositis have not been identified. Here, we describe a rare case of dropped-head syndrome due to localized idiopathic inflammatory myopathy (IIM) in the splenius capitis (neck extensor) muscle where major histocompatibility complex (MHC) class I expression was up-regulated in involved muscle fibers. Interestingly, the adjacent trapezius muscle was not affected, corresponding to muscle biopsy findings that did not show any sign of inflammation or MHC class I expression. Our case report therefore suggests that selection of affected muscle in IIM might be influenced by the MHC class I expression of the muscle.

Sex differences and the roles of sex steroids in apoptosis of sexually dimorphic nuclei of the preoptic area in postnatal rats.

The brain contains several sexually dimorphic nuclei that exhibit sex differences with respect to cell number. It is likely that the control of cell number by apoptotic cell death in the developing brain contributes to creating sex differences in cell number in sexually dimorphic nuclei, although the mechanisms responsible for this have not been determined completely. The milieu of sex steroids in the developing brain affects sexual differentiation in the brain. The preoptic region of rats has two sexually dimorphic nuclei. The sexually dimorphic nucleus of the preoptic area (SDN-POA) has more neurones in males, whereas the anteroventral periventricular nucleus (AVPV) has a higher cell density in females. Sex differences in apoptotic cell number arise in the SDN-POA and AVPV of rats in the early postnatal period, and an inverse correlation exists between sex differences in apoptotic cell number and the number of living cells in the mature period. The SDN-POA of postnatal male rats exhibits a higher expression of anti-apoptotic Bcl-2 and lower expression of pro-apoptotic Bax compared to that in females and, as a potential result, apoptotic cell death via caspase-3 activation more frequently occurs in the SDN-POA of females. The patterns of expression of Bcl-2 and Bax in the SDN-POA of postnatal female rats are changed to male-typical ones by treatment with oestrogen, which is normally synthesised from testicular androgen and affects the developing brain in males. In the AVPV of postnatal rats, apoptotic regulation also differs between the sexes, although Bcl-2 expression is increased and Bax expression and caspase-3 activity are decreased in females. The mechanisms of apoptosis possibly contributing to the creation of sex differences in cell number and the roles of sex steroids in apoptosis are discussed.

Effect of matrix metalloproteinase-3 functional SNP on serum matrix metalloproteinase-3 level and outcome measures in Japanese RA patients.

OBJECTIVE: A bi-allelic polymorphism on the promoter region, -1612 ins/del A, was found to influence the production of MMP-3. Since MMP-3 plays a particularly pivotal role in joint destruction, the MMP-3 gene is thought to be an interesting target gene of disease severity in RA. We attempt to determine whether the MMP-3 promoter polymorphism is associated with serum titre of MMP-3, disease activity and severity in Japanese RA patients. METHODS: DNA samples were obtained from 1504 RA patients as part of the Institute of Rheumatology Rheumatoid Arthritis observational cohort study. From the 2006 spring data, serum MMP-3 levels of 820 patients were available by enzyme immunoassay. Joint damage score at 5-yr disease duration could be measured using the Sharp/van der Heijde method in 162 patients. Genotyping of -1612 ins/del A was performed using fluorescent-labelled fragment analysis. Differences in serum MMP-3 level and joint damage score among genotypes of -1612 ins/del A polymorphism were analysed by linear regression analysis. RESULTS: No significant differences were found among MMP-3 genotypes on patient characteristics including disease activity score (P = 0.51) or health assessment questionnaire (P = 0.99). A significant effect of risk allele on serum MMP-3 level was observed (P = 0.038), while no significant effect was observed on radiographic joint damage (P = 0.47). CONCLUSION: We conclude that MMP-3 functional polymorphism is associated with serum MMP-3 titre, but is not a direct predictor for outcome measures in Japanese RA patients.

Toluene induces rapid and reversible rise of hippocampal glutamate and taurine neurotransmitter levels in mice.

Toluene, a widely used aromatic organic solvent, has been well characterized as a neurotoxic chemical. Although the neurobehavioral effects of toluene have been studied substantially, the mechanisms involved are not clearly understood. Hippocampus, which is one of the limbic areas of brain associated with neuronal plasticity, and learning and memory functions, may be a principal target of toluene. In the present study, to establish a mouse model for investigating the effects of acute toluene exposure on the amino acid neurotransmitter levels in the hippocampus, in vivo microdialysis study was performed in freely moving mice after a single intraperitoneal administration of toluene (150 and 300 mg/kg). Amino acid neurotransmitters in microdialysates were measured by a high performance liquid chromatography system. The extracellular levels of glutamate and taurine were rapidly and reversibly increased within 30 min after the toluene administration in a dose-dependent manner and returned to the basal level by 1h. Conversely, the extracellular level of glycine and GABA were stable, and no significant change was observed after the toluene administration. To further investigate the brain toluene level in the hippocampus of toluene-administered mice, we used a solid-phase microextraction (SPME) method and examined the time course changes of toluene in the hippocampus of living mice. The brain toluene level reached the peak at 30 min after injection and returned to the basal level after 2h. In the present study, we observed the relationship between brain toluene levels and amino acid neurotransmitter glutamate and taurine levels in the hippocampus. Therefore, we suggest that toluene may mediate its action through the glutamatergic and taurinergic neurotransmission in the hippocampus of freely moving mice.

Effect of non-steroidal anti-inflammatory ophthalmic solution on intraocular pressure reduction by latanoprost in patients with primary open angle glaucoma or ocular hypertension.

AIM: To investigate the effects of a non-steroidal anti-inflammatory drug (NSAID) ophthalmic solution on latanoprost induced intraocular pressure (IOP) reduction in glaucoma patients. METHODS: Examination was conducted on 16 eyes of 16 glaucoma patients who had been given only latanoprost for at least 6 weeks. The NSAID ophthalmic solution, sodium 2-amino-3-(4-bromobenzoyl) phenylacetate sesquihydrate, was additionally given for 12 weeks into one eye (NSAID group), while sodium hyaluronic acid ophthalmic solution was administered into the other eye (control group) in a double masked fashion. The IOP measurement was performed before the start of additional administration of ophthalmic solutions, 2, 4, 6, 8, 10, and 12 weeks after the start of additional administration, and 2, 4, and 6 weeks after discontinuing additional administration. RESULTS: No significant difference was observed in the IOPs before additional administration of ophthalmic solution between the NSAID group and the control group. Following the additional administration of ophthalmic solution, IOP in the NSAID group was consistently higher than that in the control group, and a maximum difference in IOP between the two groups was 1.08 (SD 1.75) mm Hg (p = 0.03). This trend was observed even after additional administration was discontinued. CONCLUSION: NSAID ophthalmic solution may partly affect IOP reduction by latanoprost.

Prediction of future scotoma on conventional automated static perimetry using frequency doubling technology perimetry.

AIM: To see if scotoma detected with frequency doubling technology (FDT) is confirmed by Humphrey field analyser (HFA) 3 years later. METHODS: Subjects were first examined with the screening C-20-1 program of FDT. The visual field was examined annually for 4 years using HFA program C30-2. The central 58 test points in HFA were assigned to one of the 17 clusters corresponding to FDT test points. Each cluster was represented as the lowest probability symbol of total deviation (TD) of the HFA test points included in the cluster. Clusters were graded normal, suspected scotoma, and scotoma depending on probability of TD-5% or more, 5%-1%, less than 1%, respectively. Relative risk (RR) of abnormality on FDT for future scotoma on HFA was estimated. RESULTS: 80 eyes of 42 patients were followed up for 4 years. While 4.0% of normal clusters of HFA with normal FDT results developed into scotoma cluster, 20.8% of normal clusters with abnormal FDT results developed into scotoma cluster with HFA at the third year. RR for future scotoma was 5.24 (95% CI, 2.75 to 10.0, p<0.05). CONCLUSION: An abnormal result in FDT shows a high risk of future scotoma on HFA after 3 years even if the original HFA perimetry showed normal results.

Finding cases of angle-closure glaucoma in clinic setting using a newly developed instrument.

PURPOSE: To validate the applicability of a newly developed, noncontact scanning peripheral anterior chamber depth analyzer (SPAC) for screening eyes at the risk of angle-closure glaucoma (ACG). SUBJECTS AND METHODS: All glaucoma patients who visited the University of Yamanashi Hospital from February through May 2003 were enrolled, except those with aphakic eye or pseudophakic eye. Of the 552 enrolled patients, 48 with ACG or narrow angles requiring laser iridotomy (LI) were categorized as patients with high-risk ACG eyes, and those with open angle were categorized as patients with control eyes. In all, 20 patients with ACG or narrow angles requiring prophylactic LI, who were followed up by an independent private ophthalmic clinic, were enrolled for threshold analysis. Nonophthalmologists measured anterior chamber depth and the averaged values of three measurements were employed for analysis. Threshold analysis and discriminant analysis were employed for determining the sensitivity and specificity of SPAC for diagnosing eyes with high-risk ACG. RESULTS: SPAC distinguished well the high-risk ACG eyes from the control eyes, and one of the most useful criteria for screening is as follows: any of the four measured points should exceed 95% confidence interval, and sensitivity and specificity should be 97.6 and 83.5%, respectively. CONCLUSION: SPAC is thought to be useful for detecting eyes at the risk of ACG by nonophthalmologists.

Usefulness of peripheral anterior chamber depth assessment in glaucoma screening.

PURPOSE: This study was conducted to determine the usefulness of peripheral anterior chamber depth assessment in angle-closure glaucoma (ACG) screening in Japanese subjects. SUBJECTS AND METHODS: The subjects were 14,779 adults 40 years old or older. Eyes having peripheral anterior chamber depth that is 1/4 the peripheral corneal thickness (van Herick's classification: grade 2) and less than 1/4 the peripheral corneal thickness (van Herick's classification: grade 1) were extracted as narrow angle eyes, and those eyes were further examined. RESULTS: Of 14,779 subjects, 923 eyes of 505 subjects were diagnosed as narrow angle eyes (3.4%). Narrow angle eyes were observed in 4.9% of female subjects and 1.9% of male subjects, indicating a significantly higher frequency in women. The percentage of narrow angle eyes increased with age. Among the narrow angle eyes, 61 eyes of 32 subjects were diagnosed with ACG suspect (6.5%). In contrast to the frequency of ACG suspect in eyes classified as grade 1, according to van Herick's classification, being 17.9%, that in eyes classified as grade 2 was significantly lower at 5.6%. CONCLUSION: Since the incidence of ACG suspect increases as the peripheral anterior chamber depth decreases, caution for the peripheral anterior chamber depth is required for the ACG screening.

A newly developed peripheral anterior chamber depth analysis system: principle, accuracy, and reproducibility.

AIM: To develop a new, non-contact system for measuring anterior chamber depth (ACD) quantitatively, and to investigate its accuracy as well as interobserver and intraobserver reproducibility. METHODS: The system scanned the ACD from the optical axis to the limbus in approximately 0.5 second and took 21 consecutive slit lamp images at 0.4 mm intervals. A computer installed program automatically evaluated the ACD, central corneal thickness (CT), and corneal radius of curvature (CRC) instantly. A dummy eye was used for investigating measurement accuracy. The effects of CT and CRC on the measurement results were examined using a computer simulation model to minimise measurement errors. Three examiners measured the ACD in 10 normal eyes, and interobserver and intraobserver reproducibility was analysed. RESULTS: The ACD values measured by this system were very similar to theoretical values. Increase of CRC and decrease in CT decreased ACD and vice versa. Data calibration using evaluated CT and CRC successfully reduced measurement errors. Intraobserver and interobserver variations were small. Their coefficient variation values were 7.4% (SD 2.3%) and 6.7% (0.7%), and these values tended to increase along the distance from the optical axis. CONCLUSION: The current system can measure ACD with high accuracy as well as high intraobserver and interobserver reproducibility. It has potential use in measuring ACD quantitatively and screening subjects with narrow angle.

Quantitative evaluation of changes in anterior segment biometry by peripheral laser iridotomy using newly developed scanning peripheral anterior chamber depth analyser.

AIM: Using the newly developed scanning peripheral anterior chamber depth analyser (SPAC), the effects of peripheral laser iridotomy (PLI) on peripheral anterior chamber depth (PACD) were determined quantitatively as was the association between PACD and chronic elevation of intraocular pressure (IOP) after PLI. METHODS: 16 eyes of 15 patients with acute primary angle closure glaucoma (PACG) attack, 14 eyes of 14 patients with narrow angle and PACG attack in their fellow eyes, and 13 eyes of seven patients with chronic angle closure glaucoma (CACG) were enrolled. The SPAC scanned the anterior ocular segment from the optical axis to the limbus and took 21 consecutive slit lamp images at 0.4 mm intervals. A computer installed program automatically evaluated the PACD and the averaged values of three measurements were employed for analysis. RESULTS: PLI significantly increased PACD and changed the iris contour from convex to flat or concave in all the enrolled eyes. The extent of the PLI induced PACD increase was enhanced with increasing distance from the optical axis. Comparing PACDs after PLI, eyes that received prophylactic PLI showed the greatest extent of PLI induced PACD increase, followed by eyes with CACG and eyes with PACG attack. The PACD of eyes with PACG attack was almost the same as that of the fellow eyes of PACG attack before prophylactic PLI. Eyes with PACG attack showed poorer IOP control after PLI than eyes with narrow angle and CACG with PLI. CONCLUSIONS: PLI significantly increases PACD and the small PLI induced opening of PACD may contribute to chronic IOP elevation after PLI.

Distributions of two chicken bombesin receptors, bombesin receptor subtype-3.5 (chBRS-3.5) and gastrin-releasing peptide receptor (chGRP-R) mRNAS in the chicken telencephalon.

Bombesin (BN)-like peptide receptors are known to be essential to the regulation of not only homeostasis, including feeding behavior, but also of emotional systems in mammal. Recently, two novel BN receptors, chicken BN-like peptide receptor subtype-3.5 (chBRS-3.5) and gastrin-releasing peptide receptor (chGRP-R), have been identified. Here, we report the localizations of these receptors' mRNAs in the chick brain through development using in situ hybridization. First, chBRS-3.5 mRNA signals were found in the dorsal ventricular ridge at embryonic day (ED) 9. Strong signals were observed in the hyperpallium accessorium, nidopallium and nucleus basorostralis pallii, and moderate signals were found in the hippocampus, cortex piriformis, hyperpallium intercalatum, area temporo-parieto-occipitalis, nucleus striae terminalis lateralis, nucleus olfactorius anterior and organum septi lateralis at ED16. This wide expression in the pallium persisted during posthatch periods. Abundant expressions in the hyperpallium, nidopallium, considered to be similar to the mammalian cortex, as well as in the hippocampus, indicate participation of these molecules in the processing of sensory information, motor function, learning and memory. Telencephalic areas devoid of chBRS-3.5 signals were the entopallium, arcopallium anterius, globus pallidus, nucleus intrapeduncularis, tuberculum olfactorius, nucleus septalis lateralis, hypothalamic and thalamic areas. In contrast to chBRS-3.5, chGRP-R mRNA signals were found in the pallidum at ED5 and 9. At ED16, chGRP-R mRNA signals were localized in the medial striatum and hypothalamus. GRP-R expression in the hypothalamic region was phylogenically conserved. Thus, chBRS-3.5 mRNA signals were distributed in a broader region and were more intense than chGRP-R mRNA. Taken together, chGRP-R and chBRS-3.5 mRNA occurred in similar regions of mammals that express GRP-R. BN/GRP-immunoreactive neurons and varicosities were found mainly in the pallium, especially in the hyperpallium accessorium and nidopallium, and this distribution coincided with that of chBRS-3.5 mRNA. This result suggests that the endogenous ligands for chBRS-3.5 were likely BN-like peptides produced in the pallium.

A three-year prospective, randomized and open comparison between latanoprost and timolol in Japanese normal-tension glaucoma patients.

PURPOSE: To compare the longitudinal effects of treatment on intraocular pressure (IOP) and visual field performance in Japanese normal-tension glaucoma (NTG) between latanoprost and timolol. PATIENTS AND METHODS: This is an open-label, randomized, study. A total of 62 NTG patients were prospectively, consecutively enrolled. All study subjects were randomly assigned to 0.005% latanoprost instillation once daily in the morning or 0.5% timolol instillation twice daily for a prospective 3-year follow-up, and underwent a routine ocular examination every month. Automated perimetry was performed every 6 months using Humphrey field analysers. Stereophotographs of optic discs were also obtained every 6 months. RESULTS: Percentage of IOP reduction or the magnitude of IOP reduction showed no intergroup differences either at any time point (13-15%). In the visual field, the estimated rate of change in the MD value (dB/year) was -0.34+/-0.17 (SE) for the latanoprost group, and -0.10+/-0.18 (SE) for the timolol group. The estimated rate of change in MD showed no significant difference from zero in both groups, and there were no statistical intergroup differences. No changes in the optic nerve head topography in the vertical cup-to-disc ratio and rim area measured by image-analysis techniques were observed in either group. There were no patients who dropped out due to the side effects of treatment regimens. CONCLUSION: Both latanoprost and timolol single treatments reduced IOP by 13-15% at their trough effects for 3 years in Japanese NTG patients; both showed similar effects on visual field performance.

Comparison of iridial pigmentation between latanoprost and isopropyl unoprostone: a long term prospective comparative study.

AIM: To compare incidence of iridial pigmentation prospectively induced by long term treatment with latanoprost and isopropyl unoprostone (hereafter, unoprostone) in Japanese patients with glaucoma. METHODS: Patients with glaucoma treated with prostaglandin (PG) related ophthalmic solutions were sequentially enrolled. Patients treated for more than 30 months with PG related ophthalmic solutions were subjected to analysis. The entry criteria were no history of intraocular surgery, laser iridotomy, and/or laser trabeculoplasty within 12 months before and after the enrolment; and no history of uveitis; no changes in antiglaucoma drugs within 6 months before and after the enrolment. Photographs of the irides were taken under the same conditions and three glaucoma specialists evaluated the iridial pigmentation with masking of patient information. The correlation of iridial pigmentation with the background factors and the reduction of intraocular pressure (IOP) before and after the treatment were investigated. RESULTS: 48 eyes in 48 patients satisfied the enrolment criteria (25 eyes in the latanoprost group, 23 eyes in the unoprostone group). At the end of the follow up period, iridial pigmentation was present in 15 patients (60.0%) in the latanoprost group and seven patients (30.4%) in the unoprostone group. The correlation between development of iridial pigmentation and age, sex, concurrent use of other ophthalmic solutions, and IOP reduction was not significant. CONCLUSIONS: The incidence of iridial pigmentation induced by latanoprost or unoprostone is high in the case of long term treatment. Iridial pigmentation did not affect PG related ophthalmic solution induced IOP reduction.

Endoscopic treatment of delayed unions and nonunions with calcium phosphate paste.

We describe a new therapeutic procedure using a bone filler in a calcium phosphate paste (bone paste) for endoscopic surgery for delayed unions and nonunions. Guided by radiography, the site of nonunion was confirmed. Two portals were created toward the nonunion site and, through each portal, the tissue around the nonunion site was released. While confirming the site of nonunion under endoscopy, scar tissue at the nonunion was excised to expose the bone. Next, the bone paste was injected into the space between bone fragments. This therapeutic modality is indicated most favorably in cases of open fractures, in cases after replantation in which soft tissue is severely damaged but one wants to preserve as much soft tissue as possible, and in cases of nonunion with satisfactory fixation but insufficient callus formation.