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Biomedical subjects

M Nagai

Publications and source records attributed to M Nagai.

At least 109 records · Page 6Linked to original sources
PubMed1999

Effect of Lauryl Alcohol on Morphology of Uniform Polystyrene-Poly(methyl methacrylate) Composite Microspheres Prepared by Porous Glass Membrane Emulsification Technique.

Fairly uniform poly(styrene)-poly(methyl methacrylate) (PST-PMMA) composite microspheres were prepared by employing an SPG (Shirasu Porous Glass) membrane emulsification technique. A mixture of PST, PMMA, and cosurfactant [lauryl alcohol (LOH)] dissolved in dichloromethane (DCM) was used as the dispersed phase, and an aqueous phase containing poly(vinyl alcohol) and sodium lauryl sulfate was used as the continuous phase. It is necessary to add LOH to obtain uniform particles with the SPG emulsification technique. The effects of the volume of LOH on the morphology of the final particles were investigated by varying the volume of LOH from 0 to 2 ml (per 1.2 g polymer). A three-component model was developed for different PMMA/PST ratios and LOH/polymer ratios, based on Sundberg's theory; and the calculation on morphology was carried out by using the three-component model. Agreement was obtained between experimental and calculated results. When 2 ml of LOH was added, it was found that LOH can engulf the polymer particles completely; a hemicore (HCP1P2P3) morphology, where PST and PMMA formed a hemisphere core inside a LOH shell, was observed when the PMMA/PST ratio was high, while core-shell-shell (CSP1P2P3) morphology, where PMMA formed a core and PST and LOH formed an inner shell and an outer shell, respectively, was observed when the PMMA/PST ratio was low. When the volume of LOH was below 1 ml (per 1.2 g polymer), however, LOH could not always engulf the inner polymer particles completely; cored hemisphere (CHSP3P2P1), core-shell-shell (CSP2P1P3), and hemicore (HCP1P2P3) morphologies were observed, depending on the PMMA/PST ratio and volume of LOH. Copyright 1999 Academic Press.

Journal Article↗
PubMed1999

Molecular pathologic analysis of the tonsil in HTLV-I-infected individuals.

Little is known about the role of the tonsils in HTLV-I infection. We performed molecular pathologic studies of tonsils in individuals positive or negative for anti-HTLV-I antibodies (HTLV-I-Ab) to clarify histologic characteristics of tonsils in HTLV-I infection. We collected tonsils and peripheral blood samples from patients who underwent tonsillectomy in a prospective manner. HTLV-I-Ab in serum was examined and presence of HTLV-I provirus was detected by polymerase chain reaction (PCR) in extracted DNA of both peripheral blood and tonsils. Histopathologic and immunohistochemical evaluations of tonsils were performed. HTLV-I seropositivity and PCR detection of HTLV-I provirus matched perfectly. Tonsil samples from seropositive individuals showed atrophy of the mantle zone and high numbers of T cells in the marginal zone compared with findings in HTLV-I-negative samples. HTLV-I provirus could be detected only from extracted DNA of extrafollicular areas. PCR in situ hybridization also showed positive signals in some mononuclear cells located in the marginal zone. There was a significant correlation between HTLV-I proviral load in tonsils and in peripheral blood. These results suggest the presence of characteristic histologic changes and deviated localization of HTLV-I-infected cells in the tonsils of individuals positive for HTLV-I.

Antibodies, Viral↗
PubMed1999

Increased levels of soluble Fas ligand in CSF of rapidly progressive HTLV-1-associated myelopathy/tropical spastic paraparesis patients.

The interaction of Fas ligand (FasL) with Fas-bearing cells induces apoptosis and contributes to the negative regulation of peripheral T-cell responses. Membrane-bound FasL is cleaved by a matrix metalloproteinase-like enzyme and converted to a soluble form (sFasL). Recent studies suggest that such sFasL can cause systemic tissue damage. Here we report that serum and CSF levels of soluble FasL (sFasL) are markedly higher in three active phase patients with HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). All of these patients showed higher sFasL levels in CSF than in serum. Although the HTLV-1 proviral load of patients showed no correlation with serum or with CSF sFasL, CSF sFasL levels of 14 HAM/TSP patients correlated with the anti-HTLV-1 antibody titer and neopterin concentration in CSF. These results indicate that sFasL mediated mechanisms may contribute to the inflammatory process and subsequent spinal tissue damage seen in HAM/TSP patients.

Aged↗
PubMed1999

UV resonance Raman studies of alpha-nitrosyl hemoglobin derivatives: relation between the alpha 1-beta 2 subunit interface interactions and the Fe-histidine bonding of alpha heme.

Human alpha-nitrosyl beta-deoxy hemoglobin A, alpha(NO)beta(deoxy), is considered to have a T (tense) structure with the low O(2) affinity extreme and the Fe-histidine (His87) (Fe-His) bond of alpha heme cleaved. The Fe-His bonding of alpha heme and the intersubunit interactions at the alpha 1-beta 2 contact of alpha(NO)-Hbs have been examined under various conditions with EPR and UV resonance Raman (UVRR) spectra excited at 235 nm, respectively. NOHb at pH 6.7 gave the UVRR spectrum of the R structure, but in the presence of inositol-hexakis-phosphate (IHP) for which the Fe-His bond of the alpha heme is broken, UVRR bands of Trp residues behaved half-T-like while Tyr bands remained R-like. The half-ligated nitrosylHb, alpha(NO)beta(deoxy), in the presence of IHP at pH 5.6, gave T-like UVRR spectra for both Tyr and Trp, but binding of CO to its beta heme (alpha(NO)beta(CO)) changed the UVRR spectrum to half-T-like. Binding of NO to its beta heme (NOHb) changed the UVRR spectrum to 70% T-type for Trp but almost R-type for Tyr. When the pH was raised to 8.2 in the presence of IHP, the UVRR spectrum of NOHb was the same as that of COHb. EPR spectra of these Hbs indicated that the Fe-His bond of alpha(NO) heme is partially cleaved. On the other hand, the UVRR spectra of alpha(NO)beta(deoxy) in the absence of IHP at pH 8.8 showed the T-like UVRR spectrum, but the EPR spectrum indicated that 40-50% of the Fe-His bond of alpha hemes was intact. Therefore, it became evident that there is a qualitative correlation between the cleavage of the Fe-His bond of alpha heme and T-like contact of Trp-beta 37. We note that the behaviors of Tyr and Trp residues at the alpha 1-beta 2 interface are not synchronous. It is likely that the behaviors of Tyr residues are controlled by the ligation of beta heme through His-beta 92(F8)-->Val-beta 98(FG5)-->Asp-beta 99(G1 )-->Tyr-alpha 42(C7) or Tyr-beta 145(HC2).

Carboxyhemoglobin↗
PubMed1999

Study on Preparation and Morphology of Uniform Artificial Polystyrene-Poly(methyl methacrylate) Composite Microspheres by Employing the SPG (Shirasu Porous Glass) Membrane Emulsification Technique.

Fairly uniform polystyrene-poly(methyl methacrylate) (PST-PMMA) composite microspheres were prepared by employing the SPG (Shirasu Porous Glass) membrane emulsification technique. PST, PMMA, and cosurfactant (lauryl alcohol, LOH) dissolved in dichloromethane (DCM) were used as a dispersed phase, and an aqueous phase containing poly(vinyl alcohol) and sodium lauryl sulfate was the continuous phase. The effects of LOH amount on the critical pressure of emulsification (Pcr), size distribution of droplets, and morphologies of final particles were investigated. It was found that Pcr decreased with increasing LOH amount because of preferential partition of LOH on the surface of the droplets in the initial stage of emulsification. When polymer concentration or PMMA/PST ratio was low, the size distribution of droplets decreased with increasing LOH amount, whereas an inverse trend was observed when both polymer concentration and PMMA/PST ratio were high. When polymer concentration was low, PST-PMMA core-shell particles always were obtained in the absence of LOH, irrespective of the PMMA/PST ratio. In the presence of LOH, however, microdomain, hemisphere, and inverted core-shell morphologies were formed as the PMMA/PST ratio decreased from 5/5 to 1/9 (g/g). When polymer concentration was high, different morphologies such as multiplet and inverted core-core-shell were observed. Theoretical calculations of morphologies were carried out, and agreement was obtained between experimental and calculated results. Copyright 1999 Academic Press.

Journal Article↗
PubMed1999

Reciprocal gene expression of osteoclastogenesis inhibitory factor and osteoclast differentiation factor regulates osteoclast formation.

Osteoblasts/stromal cells support the formation of osteoclast-like cells (OCL) from osteoclast progenitor cells via expressing a membrane-associated protein, osteoclast differentiation factor (ODF), in the presence of osteotropic factors, whereas the cells secrete a substantial amount of osteoclastogenesis inhibitory factor (OCIF) in the unstimulated state. There are both OCL formation-supporting and the nonsupporting cell lines in osteoblasts/stromal cell lineages. The mechanism that divides osteoblasts/stromal cell lines into the two types is not known. The present study reports that OCL formation-supporting cell line ST2 showed a greatly increased level of ODF mRNA, whereas their OCIF mRNA was drastically diminished in the presence of 1alpha, 25(OH)2-dihydroxyvitamin D3 or prostaglandin E2. In contrast, MC3T3-E1 cells lacking OCL formation-supporting ability did not show a decrease in OCIF mRNA in response to the factors, despite a similar increase in ODF mRNA as ST2 cells. However, inactivated MC3T3-E1 cells secreting nothing supported OCL formation in coculture with human promyelocytic cells, HL60. On the contrary, ST2 cells did not support OCL formation from HL60 cells when cocultured in medium conditioned by 1alpha, 25(OH)2 vitamin D3-treated MC3T3-E1. These findings indicate that reciprocal gene expression of ODF and OCIF in osteoblasts/stromal cells is essential for supporting OCL formation.

Animals↗
PubMed1999

HLA alleles determine human T-lymphotropic virus-I (HTLV-I) proviral load and the risk of HTLV-I-associated myelopathy.

The risk of disease associated with persistent virus infections such as HIV-I, hepatitis B and C, and human T-lymphotropic virus-I (HTLV-I) is strongly determined by the virus load. However, it is not known whether a persistent class I HLA-restricted antiviral cytotoxic T lymphocyte (CTL) response reduces viral load and is therefore beneficial or causes tissue damage and contributes to disease pathogenesis. HTLV-I-associated myelopathy (HAM/TSP) patients have a high virus load compared with asymptomatic HTLV-I carriers. We hypothesized that HLA alleles control HTLV-I provirus load and thus influence susceptibility to HAM/TSP. Here we show that, after infection with HTLV-I, the class I allele HLA-A*02 halves the odds of HAM/TSP (P < 0.0001), preventing 28% of potential cases of HAM/TSP. Furthermore, HLA-A*02(+) healthy HTLV-I carriers have a proviral load one-third that (P = 0.014) of HLA-A*02(-) HTLV-I carriers. An association of HLA-DRB1*0101 with disease susceptibility also was identified, which doubled the odds of HAM/TSP in the absence of the protective effect of HLA-A*02. These data have implications for other persistent virus infections in which virus load is associated with prognosis and imply that an efficient antiviral CTL response can reduce virus load and so prevent disease in persistent virus infections.

Alleles↗
PubMed1999

Schizophrenia and the serotonin-2A receptor promoter polymorphism.

Serotonin-2A (5-HT2A) receptors have received much investigative attention in schizophrenia because (1) several studies have shown a decrease in the number of 5-HT2A receptors in the prefrontal cortex of postmortem brains of schizophrenic patients; (2) atypical antipsychotic drugs are antagonists for 5-HT2A receptors; and (3) a positive association between a T to C polymorphism at position 102 of the 5-HT2A receptor gene and schizophrenia has been reported. A G to A polymorphism at position -1438 of the 5-HT2A receptor gene was studied in 119 schizophrenic patients and 106 healthy control subjects, all of whom were Japanese. The genotype and allele frequencies did not differ between the patients and control subjects. Furthermore, the genotype frequency did not differ according to diagnostic subtype, family history, age at onset of illness, or daily dosage of antipsychotic medication. Our results suggest that the polymorphism does not contribute to the etiology or clinical characteristics of schizophrenia. However, the gene is greater than 20 kbp in length, and thus it is possible that other areas that affect expression of the gene may vary. We found that the -1438G/A variant was in linkage disequilibrium with the T102C polymorphism.

Adult↗
PubMed1999

Immunogenicity of Actinobacillus ApxIA toxin epitopes fused to the E. coli heat-labile enterotoxin B subunit.

Peptides KDYGASTGSSL (Epil). SLLRRRRNGEDVSV (Epi3) and DDEIYGNDGHP (Epi6), predicted to constitute immunogenic epitopes of the hemolysin-cytotoxin ApxIA of Actinobacillus pleuropneumoniae were inserted into a surface-exposed loop of the B subunit of the E. coli heat-labile enterotoxin (EtxB). The resulting chimeric proteins were recognized by monospecific antibodies against purified native ApxI and by convalescent sera of pigs that were positive for A. pleuropneumoniae serotype 1. Mice anti-sera against chimeric proteins EtxB::ApxIAEpi3 and EtxB::ApxIAEpi6 reacted with purified ApxI. These results indicate that Epi3 and Epi6 regions constitute linear epitopes of the structural ApxIA protein toxin. Epitope Epi6 which is located in the structure of the glycine rich repeats in ApxI elicits the formation of hemolysin neutralizing antibodies when introduced into mice in the form of a chimeric EtxB fusion protein. We suggest that fusion of peptide sequences to EtxB is a useful tool for the analysis of epitopes of complex proteins such as RTX toxins.

Actinobacillus pleuropneumoniae↗
PubMed1999

Apolipoprotein E epsilon 4 allele and Japanese late-onset depressive disorders.

BACKGROUND: Several studies have suggested that late-onset depressive disorder (LOD) and the apolipoprotein E (Apo E) epsilon 4 allele are associated with dementia, respectively. The Apo E polymorphism is significantly heterogeneous among races. We hypothesized that the Apo E epsilon 4 allele frequency is elevated in Japanese LOD. METHODS: The Apo E genotype was studied in 134 patients (male, 53; female, 81) with early-/late-onset depressive disorder and 105 healthy normal controls (male, 41; female, 64). The patients were subdivided into those with early onset and late onset using 45 and 50 years as the cutoff ages. All the subjects were Japanese. RESULTS: There was statistically no difference between normal control subjects and patients with depressive disorders in Apo E genotype or allele frequency. There was statistically no difference in the age of onset of depressive disorders according to the Apo E genotype. There was no relation between the age of onset of depressive disorder and the number of epsilon 4 alleles the patient had. There was also no association between early-/late-onset depressive disorder and the Apo E genotype or allele frequency. CONCLUSIONS: Our results suggest that there is no association between the Apo E epsilon 4 allele and Japanese LOD.

Adult↗
PubMed1999

Quaternary structure sensitive tyrosine residues in human hemoglobin: UV resonance raman studies of mutants at alpha140, beta35, and beta145 tyrosine.

Recent studies noted the contribution of alpha42Tyr to the T-R-dependent UV resonance Raman (UVRR) spectral changes of HbA [Nagai, M., et al. (1996) J. Mol. Struct. 379, 65-75; Huang, S., et al. (1997) Biochemistry 36, 6197-6206], but the observed UVRR changes of the Tyr residue cannot be fully interpreted with alpha42Tyr alone. To identify the remaining contributions, the 235 nm-excited UVRR spectra of Tyr mutant Hbs at alpha140, beta35, and beta145 were investigated here. The Fe-His stretching mode demonstrated that all of these mutant Hbs take the T structure in the deoxy form under these experimental conditions. The UVRR change of the Trp residue of these mutants upon the T-R transition was the same as that in HbA, indicating that the T-R-dependent UVRR change of beta37Trp is not due to stacking with Tyr residues but is due to the formation or destruction of a hydrogen bond. The recombinant Hbs beta35Tyr --> Phe and beta35Tyr --> Thr both exhibited UVRR spectra identical with that of HbA, meaning that beta35Tyr is not responsible. In the spectra of des(beta146His,beta145Tyr)Hb with inositol hexaphosphate, the frequency shift of the Tyr RR bands was the same as that in HbA but the intensity enhancement in the CO form was small, suggesting that beta145Tyr contributes to a part of the intensity change, but scarcely relates to the frequency shift. In the spectra of Hb Rouen (alpha140Tyr --> His), the frequency shifts of bands at 1617 (Y8a) and 1177 (Y9a) cm-1 following ligation were half of those in HbA, while the intensity enhancement was not detected. This result means that alpha140Tyr is responsible for both the frequency shift and the intensity changes. It is suggested that the frequency shift of the Tyr RR bands upon the T --> R transition is due to changes in the hydrogen bonding state of alpha42- and alpha140Tyr and that the intensity enhancement is due to changes in the environment of the penultimate Tyr in both alpha and beta subunits (alpha140 and beta145). These alterations in the vibrational spectra clearly demonstrate which tyrosine residues are involved in the T-R transition as a result of modification of their local environments.

Amino Acid Substitution↗
PubMed1999

Decreased expression of p33ING1 mRNA in lymphoid malignancies.

The ING1 is a newly cloned putative tumor-suppressor gene, which is involved in the p53 signaling pathway. We found decreased expression of ING1 mRNA in 4 of 5 T-cell lines and 5 of 11 B-cell lines including two Burkitt lymphomas and two myelomas. These observations suggest that decreased ING1 expression might play an important role in the development or progression of some lymphoid tumors. Polymerase chain reaction-SSCP and sequencing analyses found neither point mutations nor small deletions in the ING1 gene, suggesting that decreased expression is due to transcriptional or post-transcriptional mechanisms.

Cell Cycle Proteins↗
PubMed1999

Myeloid- and lymphoid-specific breakpoint cluster regions in chromosome band 13q14 in acute leukemia.

Abnormalities of chromosome band 13q14 occur in hematologic malignancies of all lineages and at all stages of differentiation. Unlike other chromosomal translocations, which are usually specific for a given lineage, the chromosomal translocation t(12;13)(p12;q14) has been observed in both B-cell and T-cell precursor acute lymphoblastic leukemia (BCP-, TCP-ALL), in differentiated and undifferentiated acute myeloblastic leukemia (AML), and in chronic myeloid leukemia (CML) at progression to blast crisis. The nature of these translocations and their pathologic consequences remain unknown. To begin to define the gene(s) involved on chromosome 13, we have performed fluorescence in situ hybridization (FISH) using a panel of YACs from the region, on a series of 10 cases of acute leukemia with t(12;13)(p12;q14) and 1 case each with "variant" translocations including t(12;13)(q21;q14), t(10;13)(q24;q14) and t(9;13)(p21;q14). In 8/13 cases/cell lines, the 13q14 break fell within a single 1.4 Mb CEPH MegaYAC. This YAC fell immediately telomeric of the forkhead (FKHR) gene, which is disrupted in the t(2;13)(q35;q14) seen in pediatric alveolar rhabdomyosarcoma. Seven of the 8 cases with breaks in this YAC were AML. In 4/13 cases, the 13q14 break fell within a 1.7-Mb YAC located about 3 Mb telomeric of the retinoblastoma (RB1) gene: all 4 cases were ALL. One case of myelodysplastic syndrome exhibited a break within 13q12, adjacent to the BRCA2 gene. These data indicate the presence of myeloid- and lymphoid-specific breakpoint cluster regions within chromosome band 13q14 in acute leukemia.

Adult↗
PubMed1999

Secondary myeloid/natural killer cell precursor acute leukemia following essential thrombocythemia.

The de novo leukemic transformation of essential thrombocythemia is a rare event, and usually associated with previous treatments. We describe a patient who received treatments with nitrosourea for long-standing essential thrombocythemia and subsequently developed extramedullary tumors, tentatively diagnosed as lymphoblastic lymphoma. Combination chemotherapy was initially successful, but relapsed with marked bone marrow involvement. Surface marker analysis revealed that the tumor cells had CD5, CD7, CD33, CD34, and CD56 antigens but lacked other T-cell, and B-cell markers. Immunogenotypical studies revealed germline configurations for both T-cell receptors and immunoglobulin genes. These clinical and phenotypical features are consistent with a myeloid/natural killer cell precursor leukemia, a recently proposed distinct clinical entity. To our knowledge, this is the first report of secondary leukemia of myeloid/ natural killer cell precursor origin, and suggest that myeloid/natural killer cell precursor might be a potent target of therapy-related leukemia.

Antigens, CD↗
PubMed1999

The effects of wrist distraction on carpal kinematics.

Changes in carpal kinematics under wrist distraction were studied in fresh cadaveric specimens. A magnetic tracking device measured kinematic motions of the scaphoid, lunate, and third metacarpal relative to the fixed radius in 3 planes of passive motion (coronal, sagittal, and "dart throwers") under progressive distraction loads. The change in percent contribution of the radiocarpal and midcarpal joints was calculated. Radiocarpal motion during extension was decreased as increasing traction was applied, but it increased with flexion. Motion of the scaphoid relative to the lunate was smaller in the oblique plane, resulting in less radiocarpal motion than in the sagittal plane. In the coronal plane, traction had little effect on radial deviation, but ulnar angulation of the scaphoid was greater with ulnar deviation of the wrist. These results suggest that different degrees of tension exist in the palmar and dorsal ligaments with the wrist under traction and during different planes of wrist motion. If wrist motion is desired during fixed traction, such as used clinically with external fixation, the dart-throwers motion (wrist extension with radial deviation and wrist flexion with ulnar deviation) appears to have the least impact on radiocarpal motion. If greater radiocarpal motion is desired, however, such as during postoperative mobilization, flexion-extension and radioulnar deviation will create more radiocarpal motion than the dart-thrower's motion.

Biomechanical Phenomena↗
PubMed1999

The origin of hypsarrhythmia and tonic spasms in West syndrome: evidence from a case of porencephaly and hydrocephalus with focal hypsarrhythmia.

We report on a 3-year-old girl with West syndrome and with focal hypsarrhythmia. The left hemisphere of the patient was virtually completely defective and continuous hypsarrhythmia was only seen in the residual right frontal cortex, where an interictal single photon emission computed tomography (SPECT) showed hyperperfusion. Despite a focal epileptic pattern, the tonic spasms were quite symmetrical. In our patient, spasms might not require the sensorimotor cortex, but the brainstem containing the descending pathways that control spinal reflexes and other infratentorial structures seem to be essential for the occurrence of spasms. This is in accordance with the result of an ictal SPECT that showed hyperperfusion of the brainstem and cerebellum. These findings suggest that hypsarrhythmia originates from cortical lesions, while subcortical structures may be primarily responsible for the tonic spasms in this patient.

Brain↗
PubMed1999

Messenger RNA level and protein localization of transforming growth factor-beta1 in experimental tooth movement in rats.

Transforming growth factor-beta1 (TGF-beta1) is known to be associated with local bone remodeling. We hypothesize that TGF-beta1 is induced in bone and regulates adaptive alveolar bone modeling during tooth movement. To test this hypothesis, we examined the steady-state mRNA level of TGF-beta1 and the protein localization of TGF-beta1 and its type II receptor (TbetaR-II) during experimental tooth movement. Maxillary right molars, M1, M2 and M3, of rats were moved palatally for 1, 3 and 7 d by a finger coil spring anchored to the upper incisors. The contralateral side was used as a control. After 3 d of tooth movement, intense immunostaining for TGF-beta1 and TbetaR-II was observed in cuboidally shaped osteoblasts in the tension zone, and in bone-resorbing osteoclasts in the compression zone. On the control side, few active osteoblasts and osteoclasts were observed. Reverse transcriptase-competitive polymerase chain reaction revealed significant increases in TGF-beta1 mRNA in alveolar bone cells on the tooth movement side on day 3 and day 7. Taken together, our data indicate that TGF-beta1 is induced during tooth movement and we propose that this may regulate adaptive alveolar bone modeling.

Alveolar Process↗