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Biomedical subjects

K Hamada

Publications and source records attributed to K Hamada.

At least 37 records · Page 2Linked to original sources

Crystallographic characterization of the radixin FERM domain bound to the cytoplasmic tail of the adhesion protein ICAM-2.

Radixin is a member of the ERM proteins, which cross-link plasma membranes and actin filaments. The FERM domains located at the N-terminal regions of ERM proteins are responsible for membrane association through direct interactions with the cytoplasmic domains of integral membrane proteins. Here, crystals of the complex between the radixin FERM domain and the full-length cytoplasmic tail (28-residue peptide) of intercellular adhesion molecule 2, ICAM-2, have been obtained. The crystals were found to belong to space group P3(1)21 or P3(2)21, with unit-cell parameters a = b = 100.44 (9), c = 99.49 (6) A, and contain one complex in the crystallographic asymmetric unit. An intensity data set was collected to a resolution of 2.60 A.

Animals↗

Molecular cloning of human squamous cell carcinoma antigen 1 gene and characterization of its promoter.

The squamous cell carcinoma antigen (SCCA) serves as a serological marker for squamous cell carcinomas. Molecular cloning of the SCCA genomic region has revealed the presence of two tandemly arrayed genes, SCCA1 and SCCA2, which are 95% identical in nucleotide sequence. SCCA1 is a papain-like cysteine proteinase inhibitor, while SCCA2 is a chymotrypsin-like serine proteinase inhibitor. We analyzed here the sequence and the promoter activity of the 5'-flanking region of the SCCA1 gene. Deletion analysis of SCCA1 and SCCA2 promoter identified a 471-bp core promoter region upstream of the transcription start site. The transcriptional activity of SCCA1 promoter was up-regulated in squamous cell carcinoma cells, compared with keratinocyte and adenocarcinoma cells. The ratios of SCCA1 to SCCA2 promoter activity in squamous cell carcinoma, keratinocyte and adenocarcinoma cells were respectively 1.6, 5.3 and 2.8. Position -50 of SCCA1 and SCCA2 promoters played an important role in determining the promoter activities of SCCA1 and SCCA2. These findings suggest that the transcriptional regulation of SCCA1 and SCCA2 might differ among squamous cell carcinoma, keratinocyte and adenocarcinoma cells, and that SCCA1 promoter might be a potential target of gene therapy for squamous cell carcinoma.

Antigens, Neoplasm↗

Inhibition of gap junctional intercellular communication in rat liver epithelial cells with transforming RNA.

Previous studies indicated that transforming RNA, derived from the 3' half of the U5 small nuclear RNA first stem structure, suppressed the secretory protein translation in vitro. Gap junctions facilitate homeostatic control of cell growth and differentiation and their dysfunction has been correlated with carcinogenesis. Here, we reported that transforming RNA directly suppressed the gap junction protein, connexin 43, translation and thereby inhibited functional gap junction function in rat epithelial cells. Together with previous data, this implies that altered expression of transforming RNA itself is a potential mechanism in inhibiting gap junction function during carcinogenesis.

Animals↗

A new method for the size-selective EXAFS of neutral free clusters.

Recently we have measured Photoelectron Photoion Coincidence (PEPICO) spectra for selenium dimer Se2 and larger species such as Se5 at the beamline BL-12C in Photon Factory. The PEPICO spectra reveal that the multiply charged ions, Se(z+), are produced as a result of de-excitation processes following the inner-shell excitation and fragmented to atomic ions owing to the Coulomb explosion. The branching ratios from the parent clusters to the daughter ions exhibit clear dependence on the size of the parent clusters. In particular, the branching ratios in the non-resonant absorption region are well reproduced by assuming that the charges are randomly distributed within the clusters before the Coulomb explosion. Based upon these findings, we propose a new method for the size-selective EXAFS of neutral free clusters by utilizing the PEPICO measurements.

Journal Article↗

Ionic Strength Effects of Electrolytes on Solubilized States of Water in AOT Reversed Micelles.

The limiting amounts of solubilized aqueous NaCl, NaNO(3), MgCl(2), and AlCl(3) in sodium 1,2-bis-(2-ethylhexyloxycarbonyl)-1-ethanesulfonate (aerosol OT)/isooctane solutions have been measured as a function of the ionic strength of the electrolytes. In any system, the limiting amounts increased up to the optimal ionic strength (Io(opt)), and afterward, as ionic strength increased, they decreased and were followed by a constant. These increased and decreased curves of water solubilization below and above Io(opt), called, respectively, salting-in and salting-out curves, could be interpreted, respectively, from the counteracting effects of attractive intermicellar interaction and interfacial bending stress. The effects of electrolytes on the solubilized states of water by micelles were examined using NMR and near-infrared spectroscopic techniques. Consequently, two types of waters, i.e., water bound directly to the ionic head groups of the surfactant in reversed micelles and water interacting with the hydrated head groups in swollen micelles through hydrogen bonds, were almost unaffected by any electrolytes, whereas much greater effects were observed for bulklike water in W/O microemulsions. The order of the degree of lowering of salting-out curves in W/O microemulsion regions in each system was found to agree with the Hofmeister series. Copyright 2001 Academic Press.

Journal Article↗

Standing stock and production rate of phytoplankton and a red tide outbreak in a heavily eutrophic embayment, Dokai Bay, Japan.

The seasonal variation of phytoplankton biomass and primary productivity in a heavily eutrophic embayment, Dokai Bay, Japan, was determined. Dokai Bay was characterized by high phytoplankton biomass and productivity during summer and low phytoplankton biomass and productivity during other seasons. The results suggested that phytoplankton growth was limited by only irradiance and water temperature under the high nutrient concentrations available for phytoplankton growth in the entire year. Moreover, in spite of sufficient nutrient for phytoplankton growth in the entire year, a red tide occurred only in the summer period in this bay. Our results suggested that a red tide occurred by the high phytoplankton growth rate in the summer season, but in other periods surface phytoplankton was flushed out of the bay before forming the red tide, because phytoplankton growth rate was low and could not form the red tide due to low irradiance and low water temperature.

Animals↗

Degradation and fracture of Ni-Ti superelastic wire in an oral cavity.

Superelastic Ni-Ti wire is widely used in orthodontic clinics, but delayed fracture in the oral cavity has been observed. Because hydrogen embrittlement is known to cause damage to Ti alloy systems, orthodontic wires were charged with hydrogen using an electro-chemical system in saline. Tensile tests were carried out, and fracture surfaces were observed after hydrogen charging. The strength of the Co-Cr alloy and stainless steel used in orthodontic treatment, was not affected by the hydrogen charging. However, Ni-Ti wire showed significant decreases in strength. The critical stress of martensite transformation was increased with increasing hydrogen charging, and the alloy was embrittled. The fractured surface of the alloys with severe hydrogen charging exhibited dimple patterns similar to those in the alloys from patients. In view of the galvanic current in the mouth, the fracture of the Ni-Ti alloys might be attributed to the degradation of the mechanical properties due to hydrogen absorption.

Absorption↗

Renal vascular walls in patients with preeclampsia superimposed on essential hypertension.

This study was performed to clarify the relationship between changes in contractile proteins in renal vascular walls and the prognosis of hypertension during pregnancy. Twenty preeclamptic patients underwent renal biopsies after delivery and were divided into the following three groups: group I, patients with persistent hypertension after delivery (n = 7; mean age, 34.8 +/- 1.4 years [SE]); group II, patients who became normotensive after delivery and hypertensive again during follow-up (n = 5; mean age, 34.8 +/- 1.6 years), and group III, patients who became normotensive after delivery (n = 8; mean age, 28.0 +/- 1.0 years). We also examined age-matched healthy controls (group IV; n = 7; mean age, 34.9 +/- 1.5 years). Renal biopsy specimens were immunohistochemically stained by the avidin-biotinylated peroxidase complex method using antimonoclonal smooth muscle cell myosin heavy chain isoform antibodies (SM-1, SM-2) and antimonoclonal alpha-smooth muscle cell actin antibody (actin). We estimated and semiquantitatively scored the degree of staining in each section. In interlobular arteries, SM-1, SM-2, and actin staining in group I were significantly reduced compared with group IV (SM-1, SM-2, P: < 0.05; actin, P: < 0.01). In afferent arterioles (Afs), SM-1, SM-2, and actin staining were reduced in group I. SM-2 staining in group I was significantly reduced compared with the other three groups (versus group II, P: < 0.05; versus groups III and IV, P: < 0.01). These findings suggest that phenotypic changes in vascular smooth muscle cells (especially the disappearance of SM-2 in Afs) reflect the stage of underlying essential hypertension and can predict from the change in hypertension during pregnancy whether it will persist after delivery.

Actins↗

Biochemical markers in the synovial fluid of glenohumeral joints from patients with rotator cuff tear.

It is known that rotator cuff tears are sometimes accompanied by joint destruction. Our purpose was to elucidate the pathology with this condition. Thirty-two synovial fluid (SF) samples aspirated from the glenohumeral joints of patients with rotator cuff tears, including 7 with partial-thickness and 25 with full-thickness tears of the rotator cuff (10 massive and 15 isolated supraspinatus tendon (SSp) tears), were examined. Collagenase (MMP-1), stromelysin 1 (MMP-3), tissue inhibitor of metalloproteinases-1 (TIMP-1) and carboxy-terminal type II procollagen peptide (pCOL Il-C) were measured in the SF using the respective sandwich enzyme immunoassays. Glycosaminoglycan (GAG) was also quantified with a cationic dye binding method using 1,9-dimethylmethylene blue. Levels of any molecules except pCOL II-C in the SF appeared to be higher in full-thickness tears than those in partial-thickness tears. Moreover, levels of MMP-1, MMP-3 and GAG in the SF were significantly higher in massive tears of the rotator cuff in comparison with those in isolated SSp tears. Such significance was not observed in the levels of TIMP-1 or pCOL II C in the SF. We examined the relation of those levels with operative findings or clinical parameters from full-thickness tears, and observed significant correlations of the tear size with the levels of MMP-1, MMP-3 and GAG in the SF. Although these marker molecules in SF do not always originate from cartilage, our results may indicate the potential for accelerated cartilage-degrading activity in the glenohumeral joint in massive tears of the rotator cuff.

Adult↗

Vascular endothelial growth factor (VEGF) expression in the subacromial bursa is increased in patients with impingement syndrome.

Vascular endothelial growth factor (VEGF), which is known to be an angiogenetic factor, plays an important role in the inflammation of synovial tissue. To investigate the relationships between VEGF and clinical symptoms in rotator cuff disease, VEGF expression was examined using RT-PCR and immunohistochemical analysis in 50 patients with this disease (26 with full-thickness cuff tear, 12 with partial-thickness tear, and 12 with subacromial bursitis). VEGF mRNA expression was detected in 40 out of 50 patients by RT-PCR. VEGF mRNA expression was found more frequently in the patients with motion pain (39 out of 41) than in those without motion pain (1 out of 9) with statistical significance (Fisher's test, P < 0.001). Thirty-one out of 33 patients with synovial proliferation showed VEGF mRNA expression, whereas the expression of this transcript was found in 9 out of 17 patients without synovial proliferation. This association with synovial proliferation was also significant (Fisher's test, P = 0.0013). Thirty out of 41 patients with motion pain had synovial proliferation but 3 out of 9 patients without motion pain had synovial proliferation. In all these 30 patients with both motion pain and synovial proliferation, VEGF mRNA expression was detected. This association between motion pain and synovial proliferation was also significant (Fisher's test, P < 0.05). The mean vessel count and area in subacromial bursa expressing VEGF was significantly higher than in those without VEGF (Mann Whitney's U test, P < 0.01). These results suggested that VEGF expression is associated with vascularity, synovial proliferation and shoulder motion pain in the rotator cuff disease.

Acromion↗

Healing of full-thickness tears of avian supracoracoid tendons: in situ hybridization of alpha1(I) and alpha1(III) procollagen mRNA.

Although clinically significant, much remains unknown about the healing of the torn rotator cuff tendon, because of unavailability of appropriate animal model. The human supraspinatus tendon faces the subacromial bursa superiorly, and the joint capsule inferiorly, while the digital flexor tendon is surrounded by the synovium. We hypothesized that the supraspinatus tendon heals by the process which is different from that observed with digital flexor tendons, in which epitenon cells migrate into the torn portion circumferentially. The avian supracoracoid tendon was adopted for this experiment because of its similarity to the human supraspinatus tendon. We developed a full-thickness tendon laceration followed by primary suture. The objective of this study was to detect localization of the responsible cells for repair of the tendon. We examined the process using histology and in situ hybridization. Starting at week 1 the peritendon cells of the bursal side proliferated and migrated into the laceration site. At week 6, the tendon stumps were continuous with new connective tissue. High-level expression of procollagen mRNA in the proliferating peritendon cells on the bursal side demonstrates to contribute to the reparative process, which progressed to the joint side. This mode of repair is different from that of the digital flexor tendon.

Animals↗

Effect of high population density environment on skin barrier function in mice.

Previous reports have demonstrated that stressful stimuli markedly influenced physiological conditions and homeostasis. In this study, we examined the influence of high population density environment on barrier function and water retention property of skin in mice. Overcrowding stress induced dramatic body weight reduction and significant increase in adrenal gland weight. Macroscopic skin appearance showed moderate exfoliation and slight wrinkle formation in the stress group. Moreover epidermis hyperplasia not concomitant with inflammatory reactions such as infiltration of immunocytes and vasodilation was observed in the skin from the stress group. Skin surface conductance in the stress group was significantly lower than in the control group. On the other hand, transepidermal water loss in the stress group increased significantly, compared to the control group. Moreover, transdermal penetration of indomethacin and nicotinic acid amide was enhanced significantly. These results suggest that overcrowding stress induced impairment of barrier function and water retention property. To elucidate the mechanism of overcrowding stress-induce dry skin, contents of ceramide and pyrrolidone carboxylic acid, which are the important compounds contributing to those functions were evaluated. Both of them in stratum corneum declined significantly in the stress group. Taken together, these results demonstrate that overcrowding stress induced dry skin, and that the impairment of barrier function and water retention property correlated with decrease in ceramide and pyrrolidone carboxylic acid.

Adrenal Glands↗

Effects of high frequency repetitive transcranial magnetic stimulation on P(300) event-related potentials.

OBJECTIVE: Auditory event-related potentials (P(300)-ERPs) were analyzed before and after repetitive transcranial magnetic stimulation (rTMS). METHODS: Two rTMS trains (10 Hz, 3 s, 100% motor threshold and 5 min interval) were delivered over the left frontal area in healthy subjects. P(300)-ERPs were recorded at 14 electrode sites on the scalp using a typical oddball paradigm before and after rTMS. The latencies and amplitudes of N(100), P(200), N(200) and P(300) were measured and compared. The directed coherence (DCOH) was estimated to demonstrate information flow between different cortical areas. RESULTS: rTMS significantly influenced P(300)-ERPs. The effects differed on the different components (P<0.001). The latency of P(300) significantly increased after stimulation, which was more obvious in the frontal and central areas. The changes in P(300) amplitude were not significant (P>0.05). The DCOH from the frontal, central, parietal and occipital areas to the temporal area was significantly higher than the DCOH from the temporal area to the former 4 areas (P<0.01). CONCLUSIONS: rTMS with the present parameters can affect P(300)-ERPs, leading to a delayed P(300) component and changes in information connections around the stimulated site. Our data suggest that rTMS may postpone neuronal activities related to cognitive processing.

Adult↗

Origin of eukaryotic cell nuclei by symbiosis of Archaea in Bacteria is revealed by homology-hit analysis.

The origin of eukaryotic cell nuclei by symbiosis of Archaea in Bacteria was proposed on the basis of the phylogenetic topologies of genes. However, it was not possible to conclude whether or not the genes involved were authentic representative genes. Furthermore, using the BLAST and FASTA programs, the similarity of open reading frame (ORF) groups between three domains (Eukarya, Archaea and Bacteria) was estimated at one threshold. Therefore, their similarities at other thresholds could not be clarified. Here we use our newly developed 'homology-hit analysis' method, which uses multiple thresholds, to determine the origin of the nucleus. We removed mitochondria-related ORFs from yeast ORFs, and determined the number of yeast orthologous ORFs in each functional category to the ORFs in six Archaea and nine Bacteria at several thresholds (E-values) using the BLAST. Our results indicate that yeast ORFs related to the nucleus may share their origins with archaeal ORFs, whereas ORFs that are related to the cytoplasm may share their origins with bacterial ORFs. Our results thus strongly support the idea of nucleus symbiosis.

Archaea↗

Multifocal micronodular pneumocyte hyperplasia and lymphangioleiomyomatosis in tuberous sclerosis with a TSC2 gene.

A 45-year-old woman with a long-standing diagnosis of tuberous sclerosis (TSC) is presented. She has multifocal micronodular pneumocyte hyperplasia (MMPH) and lymphangioleiomyomatosis (LAM) of the lung, together with the detection of TSC2 gene mutation. During surgery for spontaneous pneumothorax, an open-lung biopsy was performed. Micronodules were well defined, measuring approximately 4 mm in diameter. These MMPHs were histologically composed of papillary proliferation of Type II pneumocytes, with positive immunoreactivity of keratin and surfactant apoprotein. The cystlike spaces, with dilatation and destruction of air spaces, were diffusely formed, and the walls were composed of the spindle cells. Such LAM showed positive immunoreactivity for HMB-45 (a monoclonal antibody specific for human melanoma) and tuberin (the gene product of TSC2). On germline mutation analysis using leukocytes of the present patient, a TSC2 gene mutation was confirmed as a deletion of G (or g) on Exon 9 by polymerase chain reaction-single-strand conformational polymorphism. However, no mutation was detected in her son. With microdissection analysis using paraffin-embedding lung tissues, LOH of the TSC2 gene preliminarily was detected in a LAM lesion but not in MMPH. It is suggested that MMPH, in addition to LAM, could be another pulmonary lesion in TSC patients and that the detection of TSC2 and/or TSC1 gene could essentially be useful for the pathogenesis of MMPH and LAM in TSC patients.

Adult↗

Impairment of skin barrier function in NC/Nga Tnd mice as a possible model for atopic dermatitis.

BACKGROUND: The pathogenesis and aetiology of atopic dermatitis (AD) remain unclear. Establishment of suitable animal models should aid elucidation of the pathogenesis and development of therapy. OBJECTIVES: We focused on biophysical and biochemical parameters in the skin of NC/Nga Tnd mice to evaluate similarities to and differences from AD. METHODS: Biophysical (transepidermal water loss and skin surface conductance) and biochemical parameters (ceramide contents and activity of ceramide-metabolizing enzymes) were measured in NC/Nga Tnd mice in which spontaneous dermatitis appeared under ambient laboratory conditions (ALC). RESULTS: Biophysical parameters suggested impairment of water retention properties and barrier function. The amount of ceramide in NC/Nga Tnd mice under ALC decreased significantly. These dermatological features resembled those of AD, as did the clinical signs and histological changes. CONCLUSIONS: The results described here and previous immunological studies on AD suggest that the NC/Nga Tnd mouse may be a suitable model for certain aspects of AD.

Amidohydrolases↗

New approach for weight reduction by a combination of diet, light resistance exercise and the timing of ingesting a protein supplement.

We have reported that ingesting a meal immediately after exercise increased skeletal muscle accretion and less adipose tissue accumulation in rats employed in a 10 week resistance exercise program. We hypothesized that a possible increase in the resting metabolic rate (RMR) as a result of the larger skeletal muscle mass might be responsible for the less adipose deposition. Therefore, the effect of the timing of a protein supplement after resistance exercise on body composition and the RMR was investigated in 17 slightly overweight men. The subjects participated in a 12-week weight reduction program consisting of mild energy restriction (17% energy intake reduction) and a light resistance exercise using a pair of dumbbells (3-5 kg). The subjects were assigned to two groups. Group S ingested a protein supplement (10 g protein, 7 g carbohydrate, 3.3 g fat and one-third of recommended daily allowance (RDA) of vitamins and minerals) immediately after exercise. Group C did not ingest the supplement. Daily intake of both energy and protein was equal between the two groups and the protein intake met the RDA. After 12 weeks, the bodyweight, skinfold thickness, girth of waist and hip and percentage bodyfat significantly decreased in the both groups, however, no significant differences were observed between the groups. The fat-free mass significantly decreased in C, whereas its decrease in S was not significant. The RMR and post-meal total energy output significantly increased in S, while these variables did not change in C. In addition, the urinary nitrogen excretion tended to increase in C but not in S. These results suggest that the RMR increase observed in S might be associated with an increase in body protein synthesis.

Adult↗

Contrasting effects of zonisamide and acetazolamide on amygdaloid kindling in rats.

PURPOSE: Zonisamide (ZNS) and acetazolamide (AZM) are two antiepileptic drugs (AEDs) that differ in clinical efficacy. To elucidate the mechanisms of action of these compounds, we investigated their therapeutic and prophylactic effects in rats by using a kindling model of partial epilepsy. METHODS: Electrodes were implanted into the left amygdala of adult male Wistar rats. The animals were stimulated at the afterdischarge threshold until five stage 5 seizures were induced. The generalized seizure threshold was then determined. Therapeutic effects were examined in rats manifesting successive convulsions with near-threshold stimulation. To test prophylactic effects, drugs were administered intraperitoneally before daily kindling stimulation until the animal had a stage 5 seizure or reached day 18. RESULTS: ZNS (10-40 mg/kg; n=6) suppressed kindled seizures in a dose-dependent manner. Repeated administration for 7 days produced tolerance to anticonvulsive effects. AZM (25-200 mg/kg; n=7) showed limited therapeutic effect, alleviating only the clonic convulsion in stage 5 seizures and reducing afterdischarge duration. Secondary generalization was not significantly suppressed during repeated treatment (50-200 mg/kg; n=6). ZNS, 25 or 40 mg/kg (n=8), significantly retarded seizure development; 15.0 or 17.0 daily stimulations were required to produce a stage 5 seizure. AZM, 50-200 mg/kg (n=6), also retarded seizure development, with 14.0-14.8 stimulations required. CONCLUSIONS: ZNS exhibited modest therapeutic and prophylactic effects, whereas AZM showed mainly prophylactic effects. Hypotheses are presented that may explain the mechanisms of action of these drugs.

Acetazolamide↗