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Biomedical subjects

Kazuhiro Yamada

Publications and source records attributed to Kazuhiro Yamada.

At least 19 recordsLinked to original sources

[Optimization of three-dimensional triple IR fast spoiled gradient recalled acquisition in the steady state (FSPGR) to decrease vascular artifact at 3.0 Tesla].

The purpose of this study was to decrease vascular artifacts caused by the in-flow effect in three-dimensional inversion recovery prepared fast spoiled gradient recalled acquisition in the steady state (3D IR FSPGR) at 3.0 Tesla. We developed 3D triple IR (3IR) FSPGR and examined the signal characteristics of the new sequence. We have optimized scan parameters based on simulation, phantom, and in-vivo studies. As a result, optimized parameters (1st TI=600 ms, 3rd TI=500 ms) successfully have produced the vessel signal at more than 40% reduction, while gray-white matter contrast was preserved. Moreover, the reduced artifact was also confirmed by visual inspection of the in-vivo images for which this condition was used. Thus, 3D 3IR FSPGR was a useful sequence for the acquisition of T1-weighted images at 3.0 Tesla.

Adult↗

[Vessel contrast and saturation effect at three Tesla in three-dimensional time of flight magnetic resonance angiography].

The purpose of this study was to clarify the effects of the increased field strength of 1.5T vs. 3.0T on blood vessel contrast and saturation in 3D time of flight MR angiography. Vessel contrast and saturation effect were evaluated for various flow velocities and flip angles using a steady-state flow phantom. The results showed increased vessel contrast and decreased saturation of blood on 3.0T compared to 1.5T, due to the following factors: 1) sufficient saturation of stationary tissue by extended T1 relaxation time, 2) increased signal-to-noise ratio (SNR), and 3) high in-flow effect. However, the blood signal on 3.0T revealed a strong tendency to phase dispersion compared to that of 1.5T. Therefore, in order to exploit the characteristic of high field strength in 3.0T, it is suggested that adequate consideration of these factors is important in setting the image acquisition parameters.

Imaging, Three-Dimensional↗

[Experimental study of coil selection considered from subject size].

We examined the index of coil selection in consideration of subject size to measure signal-to-noise ratio (SNR) and uniformity in a phantom experiment. The QD-Head, 8-channel-Head, and 8-channel-Head-Neck coil were used for head examination, QD-Body, 4-channel-TORSO, and 8-channel-Body coil were used for body examination. In the examination of the head, measurements were performed at various positions in a coil. The SNR measured by the QD-Head coil showed the highest values for 9 cm or 12 cm diameters of the phantom, and 10 cm to 15 cm distance from the coil entrance. The QD-Head coil also showed adequate uniformity at most sizes of phantoms. In the body examination, both multi-channel coils showed an SNR higher than the QD-Body coil for all sizes, but they were inferior to the QD-Body coil in uniformity. The 4-channel-TORSO coil showed an SNR higher than the 8-channel-Body coil when a phantom with a diameter of 21 cm or less was used. Since sensitivity distributions differ according to various cases, the 8-channel coil does not necessarily always show a high SNR. It is necessary to perform coil selection according to the subject's size and purpose.

Magnetic Resonance Imaging↗

Human methionine synthase reductase is a molecular chaperone for human methionine synthase.

Sustained activity of mammalian methionine synthase (MS) requires MS reductase (MSR), but there have been few studies of the interactions between these two proteins. In this study, recombinant human MS (hMS) and MSR (hMSR) were expressed in baculovirus-infected insect cells and purified to homogeneity. hMSR maintained hMS activity at a 1:1 stoichiometric ratio with a K(act) value of 71 nM. Escherichia coli MS, however, was not activated by hMSR. Moreover, hMS was not significantly active in the presence of E. coli flavodoxin and flavodoxin reductase, which maintain the activity of E. coli MS. These results indicate that recognition of MS by their reductive partners is very strict, despite the high homology between MS from different species. The effects of hMSR on the formation of hMS holoenzyme also were examined by using crude extracts of baculovirus-infected insect cells containing hMS apoenzyme (apoMS). In the presence of MSR and NADPH, holoenzyme formation from apoMS and methylcobalamin was significantly enhanced. The observed stimulation is shown to be due to stabilization of human apoMS in the presence of MSR. Apoenzyme alone is quite unstable at 37 degrees C. MSR also is able to reduce aquacobalamin to cob(II)alamin in the presence of NADPH, and this reduction leads to stimulation of the conversion of apoMS and aquacobalamin to MS holoenzyme. Based on these findings, we propose that MSR serves as a special chaperone for hMS and as an aquacobalamin reductase, rather than acting solely in the reductive activation of MS.

5-Methyltetrahydrofolate-Homocysteine S-Methyltran↗

Can the presence of carotid artery calcification on panoramic radiographs predict the risk of vascular diseases among 80-year-olds?

OBJECTIVE: The purpose of this study was to evaluate whether subsequent vascular diseases and related death could be predicted by the presence of carotid artery calcification detected on panoramic radiographs among elderly persons. STUDY DESIGN: We evaluated 659 panoramic radiographs from 262 male and 397 female 80-year-old subjects and their general medical examination data to assess the relationship between the presence of carotid artery calcification and vascular disease risk at baseline examination. Of these subjects, the occurrence of vascular diseases within 5 years after baseline examination was examined in 191 subjects. Further, the causes of death were examined in 108 subjects who died within 5 years after baseline examination. RESULTS: There was a significant difference in the history of past vascular diseases among subjects with and without carotid artery calcifications; however, no significant difference in the occurrence of subsequent vascular diseases was found among them. Further, there was no significant difference in the occurrence of vascular disease-related death within 5 years after baseline examination among subjects with and without carotid artery calcifications (P = 0.719). CONCLUSION: Our results suggest that the presence of carotid artery calcifications on panoramic radiographs may be related to the history of past vascular diseases; however, this is not a useful marker for subsequent vascular diseases and related death among 80-year-olds.

Aged, 80 and over↗

Protein complex expression by using multigene baculoviral vectors.

Elucidation of the molecular basis of protein-interaction networks, in particular in higher eukaryotes, is hampered by insufficient quantities of endogenous multiprotein complexes. Present recombinant expression methods often require considerable investment in both labor and materials before multiprotein expression, and after expression and biochemical analysis these methods do not provide flexibility for expressing an altered multiprotein complex. To meet these demands, we have recently introduced MultiBac, a modular baculovirus-based system specifically designed for eukaryotic multiprotein expression. Here we describe new transfer vectors and a combination of DNA recombination-based methods, which further facilitate the generation of multigene cassettes for protein coexpression (Fig. 1), thus providing a flexible platform for generation of protein expression vectors and their rapid regeneration for revised expression studies. Genes encoding components of a multiprotein complex are inserted into a suite of compatible transfer vectors by homologous recombination. These progenitor constructs are then rapidly joined in the desired combination by Cre-loxP-mediated in vitro plasmid fusion. Protocols for integration of the resulting multigene expression cassettes into the MultiBac baculoviral genome are provided that rely on Tn7 transposition and/or Cre-loxP reaction carried out in vivo in Escherichia coli cells tailored for this purpose. Detailed guidelines for multigene virus generation and amplification, cell culture maintenance and protein production are provided, together with data illustrating the simplicity and remarkable robustness of the present method for multiprotein expression using a composite MultiBac baculoviral vector.

Baculoviridae↗

Analysis of head and mandibular tapping movements in patients with mandibular protrusion.

The present study was conducted to investigate whether there was a functional coupling between the head and mandibular movements in ten patients with mandibular protrusion (MP) and ten control subjects with normal occlusion (Normal), using a six degrees-of-freedom measuring device. Single-peak waveforms were predominantly seen in both MP (98.2%) and Normal (99.3%). However, vertical displacements of the upper and lower incisor points (VD(UIP) and VD(LIP)) were all significantly larger in MP than those in Normal. The ratio VD(UIP)/VD(LIP) also increased more sharply with an increase in VD(LIP) in MP, compared to that in Normal. Mandibular rotation in MP was also significantly larger than that in Normal. The results showed that, in MP, the head moves more vertically in rhythmical coordination with mandibular movement during tapping. Finally, it may be that this larger vertical head movement is related to the greater condylar rotation in MP subjects.

Adolescent↗

Autoimmunity against neurofilament protein and its possible association with HLA-DRB1*1502 allele in glaucoma.

Glaucoma is understood as a neurodegenerative disease and intraocular pressure has been regarded as the major risk factors for the optic nerve damages. However, recent studies suggested that several risk factors including autoimmunity are also shown to play important roles in glaucoma. To identify the retinal antigen in glaucoma, we used the serological analysis of recombinant cDNA expression libraries (SEREX) approach and quantified IgG antibodies directed against the identified antigens in an ELISA. We identified neurofilament protein and the prevalence of anti-bovine neurofilament light subunit (NF-L) autoantibodies in glaucomatous patients was significantly higher than in healthy controls and patients with other uveitic and optic nerve diseases (P<0.05). In addition, our immunogenetic analysis showed a possible association between HLA-DRB1*1502 allele and the patients positive for anti-NF-L autoantibodies. It suggests that the HLA class II-linked gene may be involved in development of autoimmunity in patients with glaucoma.

Adult↗

Regulation of human methylenetetrahydrofolate reductase by phosphorylation.

Methylenetetrahydrofolate reductase (MTHFR) catalyzes the reduction of methylenetetrahydrofolate to methyltetrahydrofolate, the methyl donor for the conversion of homocysteine to methionine. Regulation of MTHFR activity is crucial for maintaining cellular concentrations of methionine and S-adenosylmethionine (AdoMet). Purified recombinant human MTHFR expressed in insect cells is multiply phosphorylated on an N-terminal extension of the protein that contains a highly conserved serine-rich region. Treatment by alkaline phosphatase removes seven phosphoryl groups from the enzyme. Thr-34 was identified as one of the seven phosphorylation sites by using a monoclonal antibody directed toward pThr-Pro. Mutation of Thr-34 to Ala completely blocks modification as judged by mass spectrometric analysis, suggesting that Thr-34 is the priming phosphorylation site. The Thr34Ala mutant was expressed in baculovirus-infected insect cells, and its enzymic properties were compared with wild-type enzyme. The mutant enzyme and alkaline phosphatase-treated wild-type enzyme are more active than untreated wild-type enzyme and less sensitive to inhibition by saturating AdoMet, indicating that phosphorylation at Thr-34 is critical for allosteric regulation of human MTHFR activity by AdoMet. The absence of methionine and the presence of adenosine in the cell culture medium, which lead to a low intracellular AdoMet/S-adenosylhomocysteine ratio, are associated with faster electrophoretic mobility of MTHFR, presumably because of less or no phosphorylation. Because the faster-mobility MTHFR is associated with the more active form of MTHFR, this response is likely to increase methionine production. Those observations suggest that AdoMet functions not only as an allosteric inhibitor but also to control phosphorylation of human MTHFR.

Alkaline Phosphatase↗

Heat, phosphorus NMR and microcalorimetry in relation to the mechanism of filament sliding.

For muscle heat measurements the methods available are sensitive and rapid, and the heat is related to the chemical changes in a manner that provides a firm outline for understanding the mechanism of contraction. For example linear dependence of the shortening heat on the sarcomere length has shown that the rate of turnover of cross-bridges increases during shortening. However, heat is bound to lack specificity. In order to cope with this problem, various methods such as rigorous chemical analyses, phosphorus NMR and microcalorimetry have been introduced. As a result of ultra-rapid freezing and chemical analysis by D. R. Wilkie (Gilbert, Kretzchmar, Wilkie and Woledge, 1971), the energy balance discrepancy between (heat + work) and the amount of phosphocreatine (PCr) split emerged, i.e. the unexplained enthalpy. Calcium ions move from the sarcoplasmic reticulum to the calcium-receptive proteins in the sarcoplasm during contraction. In an attempt to find the cause of the unexplained enthalpy, microcalorimetry of calcium binding to calcium-receptive proteins has been performed. The results have shown that calcium ions dislocated between sites within the sarcoplasm on activation may produce about 1/3 of the unexplained heat. In addition calcium pump should operate by consuming PCr to relocate the calcium after the contraction. Time-resolved phosphorus NMR has also shown that a certain amount of PCr splitting continues during early minute of recovery period after the contraction without Pi released. This delayed splitting of PCr is most likely caused by the kinetic properties of the contractile proteins and can explain another 1/3 of the unexplained enthalpy. The mechanism of how muscle is regulated is another important question. Studies of calcium binding to calcium-receptive proteins in the sarcoplasm by using titration microcalorimetry has shown that troponin C has a characteristic single calcium-binding site that is most likely to be involved in the regulation of contraction.

Actin Cytoskeleton↗

Double contours and craniofacial morphology.

This study was done to try to clarify the relationship between double contours and craniofacial morphology. The study sample included 56 pre-orthodontic patients with signs and symptoms of temporomandibular joint (TMJ) disorders. A comparison of craniofacial structures was done on 32 subjects with bilateral condylar bone change (BBC group: 28 female and 4 male) and 24 subjects with no condylar bone change (NBC group: 21 female and 3 male). The BBC showed significantly greater antegonial and ramus notch depths, as well as significantly more retruded mandibles, shorter ramus heights, and larger mandibular plane angles than the NBC. BBC subjects with bilateral double contours showed significantly more retruded mandibles, larger antegonial notch depth, and shorter lengths from the Sella to the Articulare than BBC subjects with no double contours. The study showed that important relationships exist between the presence of double contours and mandibular morphology and TMJ position.

Adolescent↗

Condylar bone change and sagittal incisal and condylar paths during mandibular protrusive excursion.

This study showed significant differences in sagittal condylar and incisal path angles during mandibular protrusive excursion, as well as flattening of the condylar path related to the existence and type of condylar bone change. Twenty-eight (28) patients with signs and symptoms of TMJ disorders were studied, using a six-degrees-of-freedom measuring device and helical CT. Sagittal incisal and condylar path angles at two mm and three mm condylar path lengths (CPL) were significantly shorter in the bilateral condylar bone change (BBC), compared to the no bone change (NBC) group. Also, NBC incremental sagittal condylar path angles from three to five mm CPL and NBC condylar path curvature at five mm CPL were both significantly larger than in BBC. Comparing types of bone change, incremental sagittal condylar path angles from three to five mm CPL was significantly less in osteophyte than in NBC or erosion groups. Condylar path curvature at five mm CPL was also significantly less in osteophyte than in NBC.

Dental Occlusion↗

Craniofacial morphology and inclination of the posterior slope of the articular eminence in female patients with and without condylar bone change.

This study investigated the association of craniofacial and glenoid fossa shapes and temporomandibular joint (TMJ) pathology in 39 orthodontic patients with signs and symptoms of TMJ disorders, using helical CT scans. Cephalometric measurements showed that 21 subjects with bilateral condylar bone change (BBC) had significantly smaller SNB angles, ramus heights and S-Ar/N-Ba ratios, as well as larger mandibular plane angles and lower anterior facial height than the 18 subjects with no condylar bone change (NBC). The average posterior slope of the left and right articular eminence in their central and lateral sections was significantly steeper in NBC than in BBC. Condylar bone change might, therefore, not only be related to the morphology of the mandible, but also of the glenoid fossa and cranial base. This appears to reflect adaptive changes in the condyle, articular eminence and cranial base in response to changes in loading.

Adolescent↗

Multivariate analysis of prognostic factors and effect of treatment in patients with IgA nephropathy.

BACKGROUND: Although the clinical and histological prognostic factors of IgA nephropathy have been investigated in detail, the value of treatment in terms of renal outcome is not well understood. METHODS: The authors examined data from 237 patients with IgA nephropathy (age 31.4+/-13.5 years, mean+/-SD) who had been followed-up for at least six months (follow-up periods, 62.3+/-45.5 months). The authors initially tested the significance of prognostic factors (age, sex, systolic blood pressure, proteinuria, serum creatinine, and histological severity) and treatment strategies (steroid therapy, renin-angiotensin system inhibitors and tonsillectomy) on renal outcome with univariate analysis, then evaluated the findings using the Cox proportional hazards model. RESULTS: Univariate and multivariate analyses showed that among the prognostic variables, a high level of serum creatinine at renal biopsy, large amounts of proteinuria, and extensive histological injury were significant risk factors for end-stage renal failure. Kaplan-Meier analysis showed that the renal survival rates associated with these factors were significantly poorer depending on their severity. Univariate analysis revealed that tonsillectomy was the only significant treatment that contributes to the maintenance of renal survival. Moreover, urinary abnormalities disappeared at a significantly higher frequency when patients were treated by tonsillectomy. The Cox proportional hazards model showed that steroid therapy independently contributed to improve renal prognosis in addition to tonsillectomy, and the hazard ratios were 0.26 (95% CI, 0.07 to 0.93) and 0.37 (95% CI, 0.14 to 0.99), respectively. CONCLUSION: Steroid therapy and tonsillectomy can independently improve renal outcome in patients with IgA nephropathy.

Adolescent↗

Effect of sevelamer on dyslipidemia and chronic inflammation in maintenance hemodialysis patients.

BACKGROUND: Hemodialysis (HD) patients often experience cardiovascular events, that might be related to altered calcium-phosphate metabolism, dyslipidemia, and chronic inflammation in addition to hypertension. Sevelamer, a non-calcium-containing phosphate binder, may improve the lipid profile of HD patients. However, the influence of sevelamer on chronic inflammation has not been clarified. METHODS: We enrolled 36 maintenance HD patients with a serum calcium (Ca) or phosphate (P) level constantly greater than 9.5 mg/dL and 5.5 mg/dL, respectively. The dose of sevelamer was titrated to achieve a serum Ca and P in the target ranges. The study period was 24 weeks. Patients underwent the following measurements: bone mineral markers, lipids, and a high-sensitivity C-reactive protein (hs-CRP). RESULTS: In the 28 patients who completed the study, sevelamer significantly reduced the mean non-high-density lipoprotein cholesterol (non-HDL-C) level by 15% and 20% (p < 0.0001) after 12 and 24 weeks, respectively, in addition to reducing the serum P level and Ca x P product. Similarly, there was a significant reduction of the serum hs-CRP level after 12 and 24 weeks [median at baseline: 1.03 mg/dL (interquartile range 0.26-3.98 mg/dL) versus 0.57 (0.17-1.47) and 0.38 (0.16-1.03), respectively, p = 0.0259]. The reduction rate of hs-CRP was significantly correlated with those of non-HDL-C (r = 0.451, p < 0.0401) and P (r = 0.453, p < 0.0008) CONCLUSION: Hs-CRP levels were reduced by sevelamer administration, as well as non-HDL-C, P, and the Ca x P product. Sevelamer may have an anti-inflammatory effect, in addition to lowering phosphate and lipid levels in HD patients.

Aged↗

[Three patients with nephrotic syndrome due to membranous nephropathy complicated by Sjögren's syndrome].

We describe three females patients (age 40, 62 and 63 y) with nephrotic syndrome due to membranous nephropathy complicated by Sjögren's syndrome(SS). The patients satisfied the criteria for SS, but not those for systemic lupus erythematosus(SLE). All of them developed nephrotic syndrome, and light immunofluorescence as well as electron microscopy of renal biopsy specimens revealed membranous nephropathy. Two were positive for the SS-A antibody, and the other was positive for the SS-B antibody. Two were complicated with primary biliary cirrhosis (PBC) and autoimmune thyroiditis, whereas the other was complicated with interstitial pneumonia, but not with either PBC or autoimmune thyroiditis. Prednisolone (PSL) with or without cyclophosphamide improved the nephrotic syndrome in two of the patients. In conclusion, some patients with SS might become complicated with nephrotic syndrome due to membranous nephropathy if another systemic autoimmune disease exists.

Adult↗