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Biomedical subjects

H Nakae

Publications and source records attributed to H Nakae.

At least 37 records · Page 2Linked to original sources
PubMed2000

Electrolyzed strong acid aqueous solution irrigation promotes wound healing in a burn wound model.

The purpose of the study was to understand the effectiveness of electrolyzed strong acid aqueous solution (ESAAS) for acceleration of epithelialization in a rat burn wound model. Eighteen anesthetized Sprague-Dawley rats received 30% total body surface area third-degree burns, and 2 days after injury, the wound eschars were removed. Rats were divided into 3 groups: Group 1, no irrigation; Group 2, irrigation with physiological saline; and Group 3, irrigation with ESAAS. Wounds were observed macroscopically until they were fully epithelialized, and epithelialized wounds were examined microscopically. Epithelialization of the wounds occurred significantly early in the rats treated with ESAAS (p < 0.05). Proliferation of associated lymphocytes and macrophages was more extensive in all rats that underwent irrigation than it was in control rats. ESAAS irrigation may promote tissue growth in burn wounds.

Acids↗
PubMed2000

Hydrogen peroxide induces midzonal heat shock protein 72 and apoptosis in sinusoidal endothelial cells of hypoxic rat liver.

OBJECTIVE: To investigate the heat shock protein (HSP) 72 expression and apoptosis induced by hydrogen peroxide in hypoxic rat liver. DESIGN: Prospective control study using the isolated rat liver. SETTING: Animal research facility. SUBJECTS: Fasted, pathogen-free specific, male Sprague-Dawley rats. INTERVENTIONS: A low-flow hypoxia model was made by reducing an afferent pressure from 10 to 2.5 cm H2O, and by perfusing the isolated rat liver for 2 hrs. MEASUREMENT AND MAIN RESULTS: We investigated the hydrogen peroxide production by using the 2'-7' dichlorofluorescein image, the induction of HSP 72 by using immunohistochemistry, and apoptosis by using terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick end-labeling method in the low flow hypoxic rat liver. In low-flow hypoxia, hydrogen peroxide production, HSP 72 expression, and apoptosis were induced in the midzone of rat liver. Prevalence of HSP 72 expression was higher in the sinusoidal endothelial cells (SEC) than in the hepatocytes. All apoptotic cells were SEC with expression of HSP 72. Hydrogen peroxide was derived from hepatocytes. Pretreatment with the specific xanthine oxidase inhibitor, sodium(-)-8-(3-methoxy-phenylsulfinylphenyl) pyrazolo [1,5-a]-1,3,5-triazine-4-olate monohydrate significantly attenuated hydrogen peroxide production, HSP 72 expression, and apoptosis of SEC in the midzone. CONCLUSION: Xanthine oxidase-dependent hydrogen peroxide induces midzonal and SEC-dominant HSP 72 expression and apoptosis in hypoxic rat liver.

Animals↗
PubMed2000

Effectiveness of electrolyzed oxidized water irrigation in a burn-wound infection model.

OBJECTIVE: The purpose of the study was to determine whether electrolyzed oxidized water (EOW) functions as a bactericide in burn injury with Pseudomonas aeruginosa infection in a rat burn-wound model. METHODS: Anesthetized Sprague-Dawley rats (n = 31) were subjected to third-degree burns to 30% of total body surface area. Two days after injury, all rats were infected with P. aeruginosa using 1 mL of a suspension containing 1 x 10(8) colony-forming units. Rats were assigned to one of three groups: no irrigation (group I), irrigation with physiologic saline (group II), or irrigation with EOW (group III). Blood culture, endotoxin levels, and survival rates were determined. RESULTS: Survival rate was significantly higher in group III than in groups I or II (p < 0.0001). Serum endotoxin levels on day 3 after infection in group III were significantly lower than the levels in group I (p < 0.01) and group II (p < 0.01). There were significant differences between the three groups in the culture of P. aeruginosa (p < 0.05). CONCLUSION: Irrigation and disinfection with EOW may become useful in preventing burn-wound sepsis.

Animals↗
PubMed2000

Long-term follow-up study of unruptured vertebral artery dissection: clinical outcomes and serial angiographic findings.

OBJECT: Although the spontaneous occurrence of an unruptured vertebral artery (VA) dissection has increasingly been recognized as a relatively common cause of stroke, and the clinical aspects of this lesion have gradually been determined, its natural course remains obscure. The main goal of this study was to clarify the management protocol for this condition by examining serial angiographic changes in patients with unruptured VA dissections. METHODS: Seventeen patients with unruptured VA dissections, including 13 men and four women, were clinically and angiographically examined between 1993 and 1998. All patients were observed using serial angiography studies. The initial angiography examinations most frequently revealed stenotic lesions (appearance of a pearl-and-string sign or string sign) in eight (47.1%) of 17 cases. In 15 cases (88.2%), changes in the lesions were evident on follow-up angiography studies. Stenotic lesions resulted in occlusion in four cases, normalization in three, and subsequent formation of an aneurysm in one case, which was treated successfully by proximal occlusion of the affected vessel performed using a detachable balloon. Occluded lesions, which were initially observed in three patients, recanalized in two patients and remained unchanged in one patient. Fusiform dilation alone was demonstrated in three patients during the initial angiography session; these lesions became normalized or were unchanged on follow-up studies. Saccular aneurysms were observed in two patients. In one of these cases, proximal ligation of the parent artery was successfully performed because of subsequent aneurysm enlargement. A double lumen, which appeared in one patient with an extradural VA dissection, became occluded. Magnetic resonance T2-weighted imaging studies revealed infarction corresponding to the posterior circulation in seven cases. During long-term observation in this series, good or excellent recovery was obtained in 14 (87.5%) of 16 patients, and moderate or severe disability in two (12.5%); one patient was lost to follow up after the second angiography study. CONCLUSIONS: A follow-up angiography study must be performed during the early stage (within approximately 3 weeks after onset of symptoms) to confirm the formation or enlargement of an aneurysm, because such conditions may be amenable to surgical treatment. Unruptured VA dissection could otherwise be treated and followed conservatively. Although the majority of dissected lesions seem likely to stabilize within a few months, as evidenced on angiography, in some cases a longer observation period is required.

Adult↗
PubMed1999

The effective removal of proinflammatory cytokines by continuous hemofiltration with a polymethylmethacrylate membrane following severe burn injury: report of three cases.

The serum levels of proinflammatory cytokines were investigated in three patients with severe burn injuries complicated by sepsis and pulmonary edema, who were treated with continuous hemofiltration (CHF) using a polymethylmethacrylate (PMMA) membrane. All patients had suffered burn injuries to more than 30% of their total body surface area (TBSA) and had burn indexes of 20 or more. Both interleukin (IL)-6 and tumor necrosis factor-alpha were detectable in one patient, while the serum IL-6 levels were elevated in the remaining two patients. The serum cytokines decreased 24 h after the initiation of CHF. Determinations of IL-6 in inflow and outflow blood samples as well as in the filtration fluid revealed that IL-6 was ultrafiltrated and/or adsorbed by the filter. Two of the three patients did not survive. Nevertheless, the results of this study indicate that since burn injuries are frequently associated with hypercytokinemia, the removal of cytokines by CHF with a PMMA membrane may be effective in the management of severe burn injuries.

Adult↗
PubMed1999

Nuclear matrix protein levels in burn patients with multiple organ dysfunction syndrome.

In this study, we investigated multiple organ dysfunction syndrome (MODS) and nuclear matrix protein (NMP), which is an indicator for apoptosis, in burn patients. The subjects were 17 patients with a total burn surface area (TBSA) of 26% or more. Eight of them had MODS, and nine did not. Seven patients died, and 10 survived. The NMP value in the group with MODS (610 +/- 318 units/ml) was significantly higher than in the group without MODS (82 +/- 31 units/ml). The NMP values of the patients who died (645 +/- 326 units/ml) were significantly higher than in the group who survived (111 +/- 95 units/ml), and they rose significantly as the number of impaired organs increased. Our findings suggested that the NMP level in burn cases associated with MODS was higher when apoptosis developed and larger numbers of organs were impaired.

Adult↗
PubMed1999

Interleukin-6 (IL-6) and IL-8 are induced in human oral epithelial cells in response to exposure to periodontopathic Eikenella corrodens.

Periodontitis is the inflammatory response in periodontal tissues elicited by bacterial colonization in periodontal pockets. In this response, pocket epithelial cells are the first cells to come into contact with bacteria. To elucidate this mechanism, we determined the adherence of the periodontopathic bacterium Eikenella corrodens 1073, which has a GalNAc-sensitive lectin-like adhesin (EcLS), to a human oral epithelial carcinoma cell line (KB) and the induction of proinflammatory cytokine production in the cells following exposure to this bacterium in vitro. In the adherence assay, EcLS played a role as the adhesin of this bacterium in adherence to KB cells. In a reverse transcriptase PCR, significant interleukin-8 (IL-8) and IL-6 mRNA levels were induced in response to exposure to this bacterium. In an enzyme-linked immunosorbent assay after an 8-h bacterial exposure, the IL-8 and IL-6 protein levels were 13.5- and 8.3-fold higher than those in the nonexposed controls, respectively. These protein responses were time dependent. Interestingly, when E. corrodens was separated from KB cells by cell culture inserts, a slight stimulation of the IL-6 and IL-8 mRNA and secreted protein levels was seen. These results imply that the direct contact of E. corrodens 1073 with oral epithelial cells is not necessarily required for the stimulation of IL-6 and IL-8 secretion. We suggest that E. corrodens induces the epithelial cells to secrete proinflammatory cytokines which serve as an early signaling system to host immune and inflammatory cells in underlying connective tissues.

Adhesins, Bacterial↗
PubMed1999

Usefulness of procalcitonin in Pseudomonas burn wound sepsis model.

Procalcitonin (PCT), a precursor of calcitonin, and endotoxin were determined in the burn wound sepsis model in which 21 Sprague-Dawley rats were scalded approximately 30% on their back. On day 2 post burn, the wounds were inoculated 1 x 10(8) colony-forming units of Pseudomonas aeruginosa. On day 5 post burn P. aeruginosa was detected by blood culture in 10 of the 21 rats (47.6%). The mortality rate 7 days after burn was 90.5%. Significant correlations were observed between serum endotoxin levels and serum PCT levels on day 5 post burn (r = 0.860, p<0.001). It was suggested that endotoxin may induce the release of PCT and that measuring the levels of PCT may be useful in diagnosing burn wound sepsis.

Animals↗
PubMed1998

Functional characteristics and the complete primary structure of ascidian gelsolin.

The body wall of the ascidian is composed of unusual multi-nucleated smooth muscle cells enriched with thin actin filaments containing troponin-tropomyosin which run along the longitudinal cell axis without being organized into striated structures. We purified an actin-binding protein of 80 kDa, tentatively termed 80K protein, from the body wall muscle of ascidian, Halocynthia roretzi, and characterized the functional properties and molecular structures. In the presence of Ca2+, the 80K protein accelerated the initial phase of actin polymerization, namely the nucleation process, decreased the level of polymerization at the steady state, caused marked reduction in viscosity of an F-actin solution, and fragmented F-actin filaments, while in the absence of Ca2+, it remained associated with F-actin without severing the filaments. The interaction of the 80K protein with actin was inhibited by phosphatidylinositol 4,5-bisphosphate (PIP2). When actin was polymerized in the presence of acrosome actin bundles from horseshoe crab sperm, the 80K protein inhibited the growth of actin filaments at the barbed end but not at the pointed end, indicating that the 80K protein functions as a barbed-end capping protein. In order to characterize the molecular structure of the 80K protein, cDNAs encoding this protein were isolated from the lambda gt11 cDNA library of the ascidian muscle by using a monoclonal antibody (AS23) specific for this protein and the entire sequence was determined. The deduced peptide sequence showed about 44% homology in amino acid residues with the human gelsolin sequence, and in addition, 6 repeating segments were observed in the sequence of the 80K protein as has been described in the gelsolin sequence. These results indicate strongly that the 80K protein belongs to the gelsolin family.

Actins↗
PubMed1998

Plasma levels of type II phospholipase A2 and nitrite/nitrate in patients with burns.

Type II phospholipase A2 (type II PLA2), nitrite/nitrate (NOx), tumor necrosis factor-alpha (TNF-alpha) and endotoxin were studied in burn patients. Type II PLA2 values and NOx values were both significantly higher in the group that died, the group with shock as a complication and the group with multiple organ dysfunction syndrome (MODS) as a complication, than in the group that survived, the group without shock as a complication and the group without MODS as a complication, respectively. The results suggested that type II PLA2 and nitric oxide (NO) may play a major role in the pathology of burn patients. Significant correlations were found between TNF-alpha and type II PLA2 values and between type II PLA2 and NOx values, suggesting the possibility of involvement in each other's production.

Adult↗
PubMed1997

Developmentally regulated mRNA splicing of clathrin assembly protein 3 (AP-3).

Clathrin assembly protein 3 (AP-3) is a neuron-specific component of clathrin coated vesicles. Because it promotes the assembly of uniform clathrin cages, AP-3 may play a regulatory role in synaptic vesicle recycling. Previously, using the monoclonal antibody MabR-18, we demonstrated that AP-3 expression starts from the embryonic stage and is maintained at high levels from the early postnatal stages through adult. In order to study the expression of AP-3 during early postnatal development at the mRNA level, RT-PCR analysis was performed. We divided the coding region of AP-3 into 10 regions and designed primers to amplify each region. As a result, developmentally regulated splicing sites were found in two regions. In one region, a PCR product with a 108-bp deletion was detected from postnatal day 10 (P10). In the other region, a product with a 15-bp deletion was increased compensating for the decrease of the undeleted product. The expression of isoforms changed mainly from around P7 to P10, whereas the level of AP-3 protein remained relatively constant throughout postnatal development. These results suggest that the expression of AP-3 isoforms with mRNA splicing is developmentally regulated in the brain and may be involved in the maturation of synaptic vesicle recycling.

Adaptor Proteins, Vesicular Transport↗
PubMed1997

An immunohistochemical study on the localization of Porphyromonas gingivalis, Campylobacter rectus and Actinomyces viscosus in human periodontal pockets.

The localization and distribution of Porphyromonas gingivalis, Campylobacter rectus and Actinomyces viscosus were studied in human periodontal pockets. After obtaining voluntary consent from 9 patients, 12 teeth and their surrounding periodontal tissue with advanced adult periodontitis were extracted carefully so as not to change the structure of the periodontal pockets. The specimens were processed into serial sections. One of the sections was stained with Brown & Brenn-modified Gram stain to observe the distribution of bacteria. The others were stained immunohistochemically by the Labelled Streptavidin Biotin method (LSAB method) using specific rabbit antibodies against selected bacteria. Some bacteria could be found within epithelial cells. P. gingivalis was found in 9/12 of the samples examined. Small aggregates of P. gingivalis were scattered in all parts of the periodontal pockets, and some of these aggregates could be seen in close contact with the epithelium. Conversely, C. rectus was observed in 5/12 of the samples examined and was predominantly located in the middle and deep pocket zones. C. rectus tended to form large clumps in both the tooth-attached and epithelium-associated plaque area. A. viscosus was observed in 7/12 of the samples examined and was localized predominantly in the tooth-attached plaque area, especially in the shallow and middle pocket zones. Although unexpected spills of unattached plaque from periodontal pockets was possible, immunohistochemical staining with species-specific antibodies was extremely sensitive and revealed the localization and the distribution of periodontal disease-associated bacteria in human periodontal pockets.

Actinomyces viscosus↗
PubMed1996

Cloning, sequencing and expression of an Eikenella corrodens gene encoding a component protein of the lectin-like adhesin complex.

A lectin-like substance (LS), that was isolated from Eikenella corrodens (Ec) 1073, migrated as proteins of about 300 and 45 kDa upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions. In this study, we cloned the gene encoding the 45-kDa protein and predicted its structure and function. Based on the N-terminal 23-amino acid (aa) sequence of this protein, we cloned the region for its N-terminus. We cloned the entire gene by means of gene walking using polymerase chain reaction and Southern hybridization. The nucleotide sequences of cloned fragments revealed an open reading frame encoding a polypeptide of 330 aa (M(r), 35748). This ORF displayed high homology to those of porins of Neisseria species. Using the T7-expression system, the 45-kDa protein was produced in E. coli. Our results suggested that the 45-kDa protein of Ec 1073 is a component of the EcLS complex, and that it is the major outer membrane protein.

Amino Acid Sequence↗
PubMed1996

Molecular analysis of the gene encoding a protein component of the Eikenella corrodens adhesin complex that is close to the carbohydrate recognition domain.

A monoclonal antibody against a lectin-like substance (LS) of Eikenella corrodens (Ec) was used for screening the Ec DNA library. Three positive clones that carried an identical 12-kb segment were obtained. A 25-kDa protein, which specifically binds to the antibody, was overproduced in all of the Escherichia coli clones. Deletion analysis showed that the gene encoding the 25-kDa protein was located within a 1.2-kb segment. The nucleotide (nt) sequence of this segment contained an open reading frame encoding a protein of 24600 Da. We purified the 25-kDa protein from the cloned E. coli strain. The sequence of the first 10 amino acids(aa) from the N-terminus of the purified 25-kDa protein agreed with that deduced from the nt sequence. Since the monoclonal antibody used in this study inhibits the physiological activity of EcLS, we concluded that the 25-kDa protein is a component of the adhesin complex, which is located near the carbohydrate recognition domain of lectin in EcLS.

Adhesins, Bacterial↗
PubMed1996

Chronological changes in the complement system in sepsis.

The time courses of serum complement levels and the severity of sepsis were compared in two groups of septic patients, one in which the patients survived (surviving group) and one in which they did not (nonsurviving group). The components of the complement system, namely, C3a, C4a, C5a, CH50, C3, C4, and C5, were measured at several points in time after the diagnosis of sepsis had been established. A 2-antibody radioimmunoassay was used to measure C3a, C4a, and C5a; the latex agglutination test was used to measure C3 and C4; nephelometry was used to measure C5; and Meyer's 50% hemolysis method was used to measure CH50. Following the diagnosis of sepsis, the levels of CH50, C3, and C4 were significantly lower in the nonsurviving than the surviving group, while the levels of C3a and C4a were significantly higher in the nonsurviving than the surviving group. The C5a levels were significantly higher in the nonsurviving than the surviving group, although no significant intergroup differences were subsequently noted. These results suggest that the serum levels of C3a, C4a, C5a, CH50, C3, and C4 could serve as indices of the severity of sepsis. Thus, monitoring the complement system may be useful for predicting the outcome of patients with sepsis.

Adult↗