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Biomedical subjects

S Hamada

Publications and source records attributed to S Hamada.

At least 91 records · Page 5Linked to original sources

Early detection of the no-reflow phenomenon in reperfused acute myocardial infarction using technetium-99m tetrofosmin imaging.

Evaluation of myocardial perfusion in the early stage of acute myocardial infarction (MI) is clinically important for adjunctive therapies to minimize infarct size. To determine the role of early scintigraphic detection of impaired myocardial reperfusion after primary coronary angioplasty (PTCA) in patients with acute MI, semiquantitative technetium-99m tetrofosmin single-photon emission tomographic (SPET) imaging was performed before primary PTCA (before; area at risk), 60 min after PTCA (after) and at 1 month (1 M; final infarct) in 35 patients with acute MI. The left ventricle was divided into 13 segments and the defect score was calculated as the sum of the perfusion defect of each segment, from 3 (complete defect) to 0 (normal perfusion). A significant myocardial perfusion change after PTCA was defined as a change in the defect score (before minus after PTCA) of >/=4. The echocardiographic asynergic score was defined as the number of asynergic (severe hypokinetic or akinetic) segments corresponding to the analogous segments on SPET images, and recovery of wall motion was calculated as absolute change in the asynergic score (before PTCA minus 1 M). Among the 35 patients, 15 (43%) had a change in the defect score of <4 (no reflow: group 1) while 20 had a change in the defect score of >/=4 (reflow: group 2). There were no significant differences between the two groups with respect to the time between admission to PTCA, revascularization time, collateral grade or Thrombolysis in Myocardial Infarction (TIMI) flow grade before PTCA. Despite the lack of a difference in area at risk between the two groups (group 1 = 12.8+/-4.3 and group 2 = 15.1+/-4.7), final infarct size in group 1 was significantly larger compared with that in group 2 (8.1+/-4.3 vs 4.9+/-3.0, P<0.001). Recovery of wall motion was significantly smaller in group 1 than in group 2 (4.3+/-1.7 to 3.5+/-1.5 vs 4. 1+/-2.1 to 1.6+/-1.6, P<0.001). In conclusion, a small change (<4) in defect score (scintigraphic no-reflow phenomenon) after primary PTCA indicates persisting impaired myocardial perfusion or irreversible cellular damage just after PTCA which is associated with poor recovery of wall motion, as compared with that observed in cases of reflow (>/=4 in defect score).

Aged↗

Regulated expression of the Shiga toxin B gene induces apoptosis in mammalian fibroblastic cells.

Shiga toxins (Stxs) produced by enterohaemorrhagic Escherichia coli may induce colonic ulceration, bloody diarrhoea and acute renal failure. The A subunit (StxA) is known to inhibit protein synthesis, whereas the B subunits (StxB) bind to Gb3 on the cell surface. However, the mechanisms by which Stxs kill target cells remain unclear. Stx1A or Stx1B genes were introduced into pcDNA3.1 vectors and transfected into NIH3T3 and HeLa cells. The Stx1B gene-transfected cells became apoptotic with accompanying DNA fragmentation, whereas the Stx1A gene-transfected cells were found to be necrotic and no DNA fragmentation occurred. The HeLa/C4 cells integrated with the Stx1B gene with a tetracycline-inducible promoter eventually produced cytoplasmic Stx1B, leading to DNA fragmentation on the addition of doxycycline. These apoptotic changes were abrogated by pretreatment with Z-VAD-fmk. These results suggest that the transfected Stx1B gene induces apoptosis by activating the caspase cascade after Stx1B expression in the cytoplasm.

3T3 Cells↗

Regulation of small intestinal transit by central nervous calcitonin receptor.

Salmon calcitonin (sCT) suppresses small intestinal transit (SIT) or motility, but the mechanism is not well understood. Bolus s. c. administration of a pharmacologic dose of sCT (140 IU/kg) to mice significantly decreased plasma calcium and phosphorus, and suppressed SIT from 1 to 8 h for plasma calcium and phosphorus or 20 h for SIT (respective maximal effects were seen at 5 h, between 2 and 8 h, and between 1 and 5 h). Significant SIT inhibition did not occur at doses smaller than 140 IU/kg. Reverse transcription-polymerase chain reactions and Southern analysis demonstrated high levels of calcitonin receptor mRNA in diencephalon and lung, moderate levels of mRNA in cerebellum, kidney, and muscle, and barely detectable amounts in cerebral cortex and thymus. No message was detectable in duodenum, jejunum, liver, testis, or heart. Specific binding of [125I] sCT was demonstrated in the diencephalon. Intracerebroventricular (i.c.v.) administration of sCT inhibited SIT time- and dose-dependently. Maximal inhibition was obtained at a dose of 4 IU/kg, 20 min after injection. Pretreatment with sCT (140 IU/kg s.c.) completely abolished inhibition of SIT by i.c.v. sCT (4 IU/kg). These results suggest that sCT binds to receptors in the central nervous system and inhibits small bowel transit.

Animals↗

Targeted salivary gland immunization with plasmid DNA elicits specific salivary immunoglobulin A and G antibodies and serum immunoglobulin G antibodies in mice.

For the development of vaccines against oral and pharyngeal pathogens invading the mucosal epithelia, both secretory and serum immunoglobulin A (IgA) and IgG antibodies and cytotoxic T lymphocytes (CTL) have been induced. We used a novel approach, targeted salivary gland (TSG) immunization, using plasmid pcDNA3/fimA, coding for Porphyromonas gingivalis fimbriae. Expression of subunit protein, fimbrillin, was observed in eukaryotic cells growing in vitro following transfection with pcDNA3/fimA. In this study, we obtained good humoral and cell-mediated immune responses in BALB/c mice by TSG administration using the above-mentioned DNA vaccine. The production of fimbria-specific IgA and IgG antibodies in saliva and serum IgG antibody was significantly stimulated by TSG immunization. Injection of DNA vaccine into salivary gland elicited high-level production of antigen-specific IgG antibody, similar to that induced following intramuscular immunization. The major IgG subclass that recognized fimbriae was IgG2a in serum from pcDNA3/fimA-immunized mice. Reverse transcription-PCR analysis of mononuclear cells from salivary glands showed that levels of Th2 cytokine-specific mRNA were higher in the immunized group than in the nonimmunized group. In addition, TSG DNA immunization resulted in the generation of antigen-specific CTL in spleen. These results indicate that TSG immunization with plasmid DNA may represent a genetic immunization strategy against infection by oral and pharyngeal pathogens that may invade local, mucosal surfaces.

Animals↗

Molecular interactions of Porphyromonas gingivalis fimbriae with host proteins: kinetic analyses based on surface plasmon resonance.

Fimbriae of Porphyromonas gingivalis are thought to play an important role in the colonization and invasion of periodontal tissues. In this study, we analyzed the interactions of P. gingivalis fimbriae with human hemoglobin, fibrinogen, and salivary components (i.e., proline-rich protein [PRP], proline-rich glycoprotein [PRG], and statherin) based on surface plasmon resonance (SPR) spectroscopy with a biomolecular interaction analyzing system (BIAcore). The real-time observation showed that the fimbriae interacted more quickly with hemoglobin and PRG than with other proteins and more intensely with fibrinogen. The significant association constant (ka) values obtained by BIAcore demonstrated that the interactions between fimbriae and these host proteins are specific. These estimated Ka values were not too different; however, the Ka values for hemoglobin (2.43 x 10(6)) and fibrinogen (2.16 x 10(6)) were statistically greater than those for the salivary proteins (1.48 x 10(6) to 1.63 x 10(6)). The Ka value of anti-fimbriae immunoglobulin G for fimbriae was estimated to be 1. 22 x 10(7), which was 6.55-fold higher than the mean Ka value of the host proteins. Peptide PRP-C, a potent inhibitor of PRP-fimbriae interaction, dramatically inhibited fimbrial association to PRP and PRG and was also inhibitory against other host proteins by BIAcore. The binding of fimbriae to these proteins was also evaluated by other methods with hydroxyapatite beads or polystyrene microtiter plates. The estimated binding abilities differed considerably, depending on the assay method that was used. It was noted that the binding capacity of PRP was strongly diminished by immobilization on a polystyrene surface. Taken together, these findings suggest that P. gingivalis fimbriae possess a strong ability to interact with the host proteins which promote bacterial adherence to the oral cavity and that SPR spectroscopy is a useful method for analyzing specific protein-fimbriae interactions.

Amino Acid Sequence↗

Secretion of functional salivary peptide by Streptococcus gordonii which inhibits fimbria-mediated adhesion of Porphyromonas gingivalis.

Porphyromonas gingivalis, a putative periodontopathogen, can bind to human salivary components with its fimbriae. We have previously shown that fimbriae specifically bind to a peptide domain shared by a major salivary component, i.e., proline-rich (glyco)proteins (PRPs). The synthetic domain peptide PRP-C (pPRP-C) significantly inhibits the fimbrial binding to PRPs. In this study, a recombinant strain of Streptococcus gordonii secreting pPRP-C was generated as a model of a possible approach to prevent the oral colonization by the pathogen. A duplicate DNA fragment (prpC) encoding pPRP-C was obtained by self-complementary annealing of synthetic oligonucleotides. prpC was connected downstream to a promoter and a gene encoding a signal peptide of Streptococcus downei glucosyltransferase I in frame. The linked fragments were inserted into the plasmid pMNK-4 derived from pVA838. The constructed plasmid was inserted to produce the transformant S. gordonii G9B, which then successfully secreted recombinant pPRP-C (r-pPRP-C) of the expected size. The concentrated bacterial culture supernatant containing r-pPRP-C inhibited the binding of P. gingivalis cells and fimbriae to PRP1 in a dose-dependent manner up to 72 and 77%, respectively. The r-pPRP-C concentrate also inhibited the coaggregation of P. gingivalis with various streptococcal strains as effectively as synthetic pPRP-C in a dose-dependent manner. Collectively, pPRP-C was found to be able to prevent P. gingivalis adherence to salivary receptor protein and plaque-forming bacteria. These results suggest that this recombination approach with a nonperiodontopathic bacterium may be suitable for the therapeutic prevention of P. gingivalis adherence to the oral cavity.

Bacterial Adhesion↗

Inhibitory effects of protamines on proteolytic and adhesive activities of Porphyromonas gingivalis.

Protamines (salmine prepared from sperm DNA of salmon and clupeine from herring sperm), which are basic peptides rich in arginine, were found to inhibit the proteolytic activity of arginine-specific cysteine protease (RC-protease) from Porphyromonas gingivalis. Lineweaver-Burk plot analysis revealed that the protamines competitively inhibited proteolytic activity with cleavage of benzoyl-L-arginine-p-nitroanilide, a synthetic substrate of RC-protease. Furthermore, the protamines were capable of binding strongly to P. gingivalis fimbriae and inhibited fimbrial interaction with immobilized fibronectin. These results clearly show that protamines are potent inhibitors of the proteolytic and adhesive activities of P. gingivalis.

Adhesins, Bacterial↗

Distribution of Porphyromonas gingivalis strains with fimA genotypes in periodontitis patients.

Fimbriae (FimA) of Porphyromonas gingivalis are filamentous components on the cell surface and are thought to play an important role in the colonization and invasion of periodontal tissues. We previously demonstrated that fimA can be classified into four variants (types I to IV) on the basis of the nucleotide sequences of the fimA gene. In the present study, we attempted to detect the four different fimA genes in saliva and plaque samples isolated from patients with periodontitis using the PCR method. Four sets of fimA type-specific primers were designed for the PCR assay. These primers selectively amplified 392-bp (type I), 257-bp (type II), 247-bp (type III), and 251-bp (type IV) DNA fragments of the fimA gene. Positive PCR results were observed with reference strains of P. gingivalis in a type-specific manner. All other laboratory strains of oral and nonoral bacteria gave negative results. The sensitivity of the PCR assay for fimA type-specific detection was between 5 and 50 cells of P. gingivalis. Clinical samples were obtained from saliva and subgingival plaque from deep pockets (>/=4 mm) of 93 patients with periodontitis. Bacterial genomic DNA was isolated from the samples, and the targeted fragments were amplified by PCR. The presence of P. gingivalis was demonstrated in 73 patients (78.5%), and a single fimA gene was detected in most patients. The distribution of the four fimA types among the P. gingivalis-positive patients was as follows: type I, 5.4%; type II, 58.9%; type III, 6. 8%; type IV, 12.3%; types I and II, 6.8%; types II and IV, 2.7%; and untypeable, 6.8%. P. gingivalis with type II fimA was detected more frequently in the deeper pockets, and a significant difference of the occurrence was observed between shallow (4 mm) and deep (>/=8 mm) pockets. These results suggest that P. gingivalis strains that possess type II fimA are significantly more predominant in periodontitis patients, and we speculate that these organisms are involved in the destructive progression of periodontal diseases.

Adult↗

Pre-operative staging of ampullary tumours by endoscopic ultrasound.

Ampullary carcinomas have a significantly higher resectability rate and better prognosis than other periampullary carcinomas, although the prognosis is poor with advanced disease. Accurate tumour staging is therefore important in surgical planning. Our objective was to evaluate the usefulness of, and problems associated with, endoscopic ultrasound (EUS) in the pre-operative staging of ampullary tumours. 35 patients with ampullary tumours were pre-operatively examined with EUS. The imaging results were compared with histopathological findings of the resected specimen according to the TNM staging classification. The overall accuracy of tumour (T) staging was 74% (26/35) for all tumours, and 67% (6/9), 71% (10/14) and 83% (10/12) respectively for T1, T2 and T3 tumours. The overall accuracy of nodal (N) staging was 63%. In diagnosing pancreatic invasion, EUS had an accuracy of 86% (30/35), a sensitivity of 83% (10/12), and a specificity of 87% (20/23). In conclusion, EUS provides an accurate method of evaluating the stage of ampullary tumours, especially infiltration into the pancreas. This modality is useful to surgeons in deciding on an appropriate therapeutic approach and in giving a prognosis.

Aged↗

Effect of mandibular advancement splint on psycho-intellectual derangements in patients with sleep apnea syndrome.

The mandibular advancement splint (MAS) was recently introduced for the management of sleep apnea syndrome (SAS), although its effects on psycho-intellectual functions have not been elucidated yet. We examined psycho-intellectual function before and after treatment with MAS in patients with SAS. Twenty patients with SAS underwent psycho-intellectual function testing before and after treatment with MAS for 3 to 4 weeks. The apnea index significantly decreased from 19.0+/-15.6 to 2.4+/-1.9. The state anxiety score significantly decreased from 44.6+/-12.1 to 33.7+/-11.1, the trait anxiety score significantly decreased from 46.2+/-13.4 to 37.6+/-13.8, and the depression scale score significantly decreased from 39.2+/-11.0 to 30.8+/-9.9 with MAS treatment. By the Cornell Medical Index and the Yatabe-Guilford test, the patients became less neurotic and less eccentric after treatment. By the Uchida-Kraepelin psychodiagnostic test, calculation ability significantly increased from 1247.4+/-402.1 to 1950.2+/-651.9. We conclude that MAS treatment reduces apneic episodes and improves psycho-intellectual derangements in patients with SAS.

Adult↗

Self-biofeedback control of heart rate during exercise.

To improve cardiac adjustment to exercise, we developed a new self-biofeedback heart rate (HR) controller. Using this device, we analyzed time courses of HR, running speed (RS), stride length (ST) and pitch of gait (PI) in response to various preset HR levels in 7 normal human subjects. When HR was preset at 80 bpm, HR increased rapidly in response to exercise and exceeded the preset level at 12. 1 s with overshoot. At the preset HRs of 100, 120, 140 and 160 bpm, the HRs increased to each preset level at 39.2, 64.5, 58.5 and 83.0 s after the onset of exercise, respectively, and the HRs were adjusted with a range of +/- 4%. For all preset HRs, RS, ST and PI increased more rapidly than the HR and reached the maximum values within 30 s. During exercise, RS, ST and PI remained constant within 1.5-5.5 min. HR, RS, ST and PI increased in proportion to the preset HR. The increases in PI against HR (DeltaPI/DeltaHR) decreased with the higher HR level, and at HRs of 160-170 bpm, HR and PI showed identical rhythm. The increases in RS were produced by 18-59% increases in PI and by 12-44% increases in ST. We concluded that, using our newly developed self-biofeedback HR control system, we could control HR to a given preset value by a change in RS due to PI and ST.

Adult↗

Differential diagnosis of lymphocytic interstitial pneumonia and malignant lymphoma on high-resolution CT.

OBJECTIVE: Lymphoproliferative disorders span a spectrum from inflammatory lesions to malignant neoplasms. The purpose of this study was to compare high-resolution CT findings of lymphocytic interstitial pneumonia with those of malignant lymphoma of the chest. MATERIALS AND METHODS: The study included 17 patients with lymphocytic interstitial pneumonia and 44 patients with malignant lymphoma (35 with non-Hodgkin's lymphoma and nine with Hodgkin's disease). Without knowledge of the pathologic diagnosis, two chest radiologists evaluated the frequency and distribution of high-resolution CT findings in both groups of patients. RESULTS: Cysts were more common in patients with lymphocytic interstitial pneumonia (14/17, 82%) than in patients with malignant lymphoma (1/44, 2%) (p < .0001). Air-space consolidation was more commonly seen in patients with malignant lymphoma (29/44, 66%) than in patients with lymphocytic interstitial pneumonia (3/17, 18%) (p < .001). Large nodules (11-30 mm in diameter) were more common in patients with malignant lymphoma (41%) than in patients with lymphocytic interstitial pneumonia (6%). Pleural effusions (25%) were seen only in patients with malignant lymphoma. We found no statistically significant difference in the distribution of lung lesions between patients with lymphocytic interstitial pneumonia and patients with malignant lymphoma. CONCLUSION: On high-resolution CT, cysts are characteristic in patients with lymphocytic interstitial pneumonia, whereas consolidation, large nodules, and pleural effusions are characteristic in patients with malignant lymphoma. These findings on high-resolution CT help differentiate lymphocytic interstitial pneumonia from malignant lymphoma.

Adult↗

A case of lymphoepithelial cyst of the pancreas.

We report a relatively rare case of lymphoepithelial cyst of the pancreas. The patient, a 43-year-old man with no subjective symptoms, was found to have a pancreatic tumor during a physical examination. Based on the ultrasonographic and abdominal computed tomographic findings, a pancreatic cystic tumor was diagnosed. Endoscopic retrograde pancreatography showed a normal duct system. Enucleation was easily performed. Macroscopically, the cyst resembled an atheroma. Histopathologic examination disclosed lymphoepithelial cyst of the pancreas.

Adult↗

[Usefulness of helical scanning CT in assessment of left ventricular papillary muscle contractility].

Three-dimensional data on the left ventricle at quasi-end-diastole and quasi-end-systole were acquired by helical scanning CT in 12 patients. Short axial images of the left ventricle and long axial images of the papillary muscle were created using multiplanar reconstruction. Left ventricular dimensions and the length of the papillary muscle were measured, and fractional shortening of the papillary muscle was calculated. There was a good correlation between left ventricular dimensions measured by echocardiogram and helical scanning CT (end-diastole: r = 0.94: end-systole: r = 0.86; fractional shortening of left ventricular dimension: r = 0.84). Papillary muscle fractional shortening was 6.3% in the ischemic group and 21.5% in the non-ischemic group (p < 0.005). Helical scanning CT is clinically useful for the assessment of left ventricular papillary muscle function.

Adult↗