Search PubMed⌕ Search

Biomedical subjects

N Hamada

Publications and source records attributed to N Hamada.

At least 109 records · Page 6Linked to original sources

Lazaroid U-74389G for 48-hour canine liver preservation.

Lazaroids have been reported to attenuate preservation and reperfusion injury. In this study, we examined whether lazaroids can improve the outcome after 48-hr canine liver preservation and transplantation. Adult female beagle dogs were randomized into 4 dosage groups (5 animals each). Lazaroid U-74389G was intravenously administered at a dose of 0 mg/kg, 6 mg/kg, 10 mg/kg, or 15 mg/kg to donors 30 min before harvesting and also to recipients 30 min before revascularization. Control animals (0 mg/kg) were given the lazaroid vehicle. The liver grafts were orthotopically transplanted after 48 hr of hypothermic preservation in UW solution. Lazaroid treatment significantly improved outcome after transplantation. Five-day animal survival increased from 0% in the control to 60% in the 6 mg/kg group, 100% in the 10 mg/kg group, and 80% in the 15 mg/kg group. Lazaroid protected the hepatocytes from damage during preservation, and enhanced energy charge and hepatic blood flow after reperfusion. Histological alterations were significantly less severe in the lazaroid-treated groups. The area of necrotic hepatocytes decreased from 43.7 +/- 17.7 in the control to 13.5 +/- 3.0 in the lazaroid 10 mg/kg group. These results indicate that lazaroid U-74389G has potential for improvement of clinical liver preservation.

Animals↗

An automated and portable low-flow pulsatile perfusion system for organ preservation.

While machine preservation reduces the incidence of delayed graft function in renal transplant recipients, it is only used in 10% of kidney transplantations. The performance of our portable, low-flow-pulsatile organ perfusion system was examined in a canine kidney autotransplantation model. Grafts were stored for 72 h by simple cold preservation in University of Wisconsin (UW) solution, or by high or low-flow machine preservation After preservation, the grafts were autotransplanted and the animals were followed for 15 days. Graft function was better in machine-preserved kidneys. Tissue biochemistry indicated that machine preservation resulted in higher levels of adenine nucleotides and better histological integrity than the cold storage. While histology and biochemistry of machine-preserved groups were similar, electromicroscopy of high-flow grafts showed mild accumulation of intravenous debris and endothelial swelling. This study shows that a simplified machine perfusion technique is effective for organ preservation.

Animals↗

Sympathetic overactivity of intraocular muscles evaluated by accommodation in patients with hyperthyroidism.

Sympathetic overactivity occurs in Graves' disease, but little is known about autonomic nervous function in the eyes of subjects with this disease. We examined this function of the intraocular muscles in 12 patients with hyperthyroid Graves' disease and 12 healthy controls. Pupil size, pupillary unrest, and accommodation were measured with a computer-assisted infrared optometer and pupillometer. The mean and the coefficient of variation of the areas of the pupils were used to express pupil size and the degree of pupillary unrest, respectively. Accommodation was measured with the target light beam moving slowly and steadily, or instantaneously, and the results are expressed as the change in the eye's refractive power in response to these movements. The mean pupil size of the patients was not different from that of the controls. Pupillary unrest in the patients was smaller than in the controls. Accommodation in the patients was lower than that of the controls. Five patients were examined again 3 months later when they became euthyroid; pupillary unrest and accommodation had improved in all five patients. There were no significant differences in the activity of sympathetic nerves governing intraocular muscles in patients with or without eyelid retraction. These results indicate that intraocular muscles are sympathetically overactive in patients with hyperthyroidism, and suggest that eyelid retraction is not caused by sympathetic overactivity alone, but by another factor or factors, in addition.

Accommodation, Ocular↗

Expression of functional Porphyromonas gingivalis fimbrillin polypeptide domains on the surface of Streptococcus gordonii.

Genetically engineering bacteria to express surface proteins which can antagonize the colonization of other microorganisms is a promising strategy for altering bacterial environments. The fimbriae of Porphyromonas gingivalis play an important role in the pathogenesis of periodontal diseases. A structural subunit of the P. gingivalis fimbriae, fimbrillin, has been shown to be an important virulence factor, which likely promotes adherence of the bacterium to saliva-coated oral surfaces and induces host responses. Immunization of gnotobiotic rats with synthetic peptides based on the predicted amino acid sequence of fimbrillin has also been shown to elicit a specific immune response and protection against P. gingivalis-associated periodontal destruction. In this study we engineered the human oral commensal organism Streptococcus gordonii to surface express subdomains of the fimbrillin polypeptide fused to the anchor region of streptococcal M6 protein. The resulting recombinant S. gordonii strains expressing P. gingivalis fimbrillin bound saliva-coated hydroxyapatite in a concentration-dependent manner and inhibited binding of P. gingivalis to saliva-coated hydroxyapatite. Moreover, the recombinant S. gordonii strains were capable of eliciting a P. gingivalis fimbrillin-specific immune response in rabbits. These results show that functional and immunologically reactive P. gingivalis fimbrillin polypeptides can be expressed on the surface of S. gordonii. The recombinant fimbrillin-expressing S. gordonii strains may provide an effective vaccine or a vehicle for replacement therapy against P. gingivalis. These experiments demonstrated the feasibility of expressing biologically active agents (antigens or adhesin molecules) by genetically engineered streptococci. Such genetically engineered organisms can be utilized to modulate the microenvironment of the oral cavity.

Animals↗

Isolation and characterization of a minor fimbria from Porphyromonas gingivalis.

We have discovered two distinctly different fimbriae expressed by the same Porphyromonas gingivalis strain. The construction of a fimA mutant of P. gingivalis ATCC 33277 has previously been reported by N. Hamada et al. (Infect. Immun. 62:1696-1704, 1994). Expression of fimbriae on the surface of the fimA mutant and the wild-type strain, ATCC 33277, were investigated by electron microscopy. The wild-type strain produced long fimbrial structures extending from the cell surface, whereas those structures were not observed on the fimA mutant. However, short fimbrial structures were seen on the surface of the fimA mutant. The short fimbrial protein was purified from the fimA mutant by selective protein precipitation and chromatography on DEAE Sepharose CL-6B. We have found that the second fimbrial structure of P. gingivalis ATCC 33277 is distinct from the 41-kDa (43-kDa) major fimbrial protein (FimA). We provisionally call this protein minor fimbriae. The molecular mass of the minor fimbriae is 67 kDa as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions after boiling at 100 degrees C. The component shows a ladder-like pattern at 80 degrees C under nonreducing conditions, suggesting a tendency to aggregate or polymerize. In immunoblotting analysis, anti-minor fimbria serum reacted with both the 100 degrees C- and the 80 degrees C-treated minor fimbriae. The anti-minor fimbria serum also reacts with the same-molecular-size fimbrial preparation from the wild-type strain. Immunogold electron microscopy showed that the anti-minor fimbria serum bound to the minor fimbria on the cell surface of the wild-type strain. This is the first report on the identification of the minor fimbria produced by P. gingivalis. These results suggest that the minor fimbriae appearing on the fimA mutant strain are produced together with numerous long major fimbriae on the wild-type strain. Moreover, the minor fimbriae are different in size and antigenicity from the earlier-reported FimA, a major 41-kDa fimbrial component of P. gingivalis.

Amino Acids↗

Changes in significance of axillary lymph node dissection for patients with breast cancer.

Axillary lymph node dissection has been a routine part of breast cancer treatment for more than 100 years. As so few patients have been shown to have positive nodes, more consideration should be given to eliminating axillary node dissection for duct carcinoma in situ (DCIS) and T1a lesions. And for patients with T1/2N0M0 cancer of the breast, lumpectomy alone without axillary dissection followed by radiation therapy of the intact breast and regional lymph nodes should be a reasonable treatment without any arm morbidity. Between September 1989 and September 1994, we have treated 40 breast cancer patients with this method and no local recurrence nor distant metastasis has been encountered so far. Therefore, it is concluded that axillary dissection should be performed routinely only for N1b lesions and larger.

Axilla↗

Glutamine metabolism of intestine grafts: influence of mucosal injury by prolonged preservation and transplantation.

The demand for glutamine increased only in the preserved intestine in the early postoperative period (3 days after transplantation). Glutamine demand of the preserved grafts returned to control and immediate levels 7 and 14 days after transplantation. Three days after intestinal transplantation, when the intestinal mucosa was actively regenerating, the demand for glutamine was markedly enhanced. The enhanced demand for glutamine was met by increased output of glutamine by the liver and skeletal muscle. Glutamine uptake by the intestinal graft was enhanced by a brief infusion of glutamine. Thus, we believe exogenous glutamine supplementation may be beneficial for the recovery of intestinal grafts with severe mucosal injury.

Animals↗

Lidocaine action and conformational changes in cytoskeletal protein network in human red blood cells.

The mechanism of action of lidocaine, which is commonly used clinically as a local anesthetic, was studied in human red blood cells. The influx of [14C]lidocaine through the cell membrane induced reversible transformation of human red blood cells from discocytes to stomatocytes. This change in shape depended on the lidocaine concentration and required both ATP and carbonic anhydrase. The lidocaine-induced shape change occurred as a result of spectrin aggregation, which altered the intracellular environment of the human red blood cells, mediated by carbonic anhydrase and activation of vacuolar type H(+)-ATPase (V-ATPase). Lidocaine controlled the influx of 22Na into the human red blood cells in a concentration-dependent manner. When incubated in media containing 6-chloro-9-[(4-diethylamino)-1-methyl-butyl]amino-2-methoxyacridine (mepacrine), an inhibitor of Na+ channels, human red blood cells changed shape from discocytes to stomatocytes and the intracellular pH decreased. This phenomenon was very similar to the shape change induced by lidocaine. These results suggest that the mode of action of lidocaine is related to a conformational change in the cytoskeletal protein network.

Adenosine Triphosphate↗

Immunohistochemical and biochemical detection of low density lipoprotein receptors in cultured rat mesangial cells.

The expression of low density lipoprotein (LDL) receptors by cultured rat mesangial cells was demonstrated using immunohistochemical and biochemical methods. Using a mouse monoclonal anti-human LDL receptor antibody, the immunofluorescence technique was applied to cultured mesangial cells and showed granular staining. Immunoblotting analysis of the membrane fraction of cultured mesangial cells showed a single band with a protein of approximately 130,000 molecular weight. In addition, [125I]LDL binding and the uptake of [125I]LDL by cultured mesangial cells were studied. Binding and uptake both reached an equilibrium after about 30 min of incubation. A 50% reduction in [125I]LDL (20 micrograms protein/ml) binding and uptake occurred when unlabelled LDL was added to cultures at 20-40 micrograms protein/ml. Addition of high density lipoprotein without apoE to the culture medium did not induce competitive inhibition of [125I]LDL binding and uptake by mesangial cells. These findings suggest that cultured mesangial cells have the capacity to express an LDL receptor that regulates the cellular uptake of LDL.

Animals↗

[The effects of sevoflurane on the coronary circulation in the instrumented dog with or without coronary stenosis].

The effects of sevoflurane on the coronary circulation were investigated in the instrumented dogs with or without coronary stenosis. A-V block was produced by injection of formalin into A-V node to control heart rate at 120 beats.min-1 by pacing and mean systemic blood pressure was held constant at 65 mmHg with the pressurized blood reservoir. Left anterior descending coronary artery was cannulated with stainless steel tube supplied from the femoral artery. Total coronary flow was measured with an electromagnetic flow probe and the regional myocardial blood flow was calculated using color microsphere technique. Coronary resistance was reduced with the increasing end-tidal concentrations of sevoflurane up to 1.2 MAC. An adjustable screw clamp was used to reduce the coronary flow down to 50% of control value. Inner/outer ratio of regional myocardial blood flow decreased with stenosis, but was unaffected with 1.2 MAC of sevoflurane. We concluded that 1.2 MAC of sevoflurane was not strong enough to dilate the coronary artery to produce the transmural steal phenomenon in the dog with coronary stenosis. Further investigations are needed to clarify how hemodynamic factors influence the transmural steal phenomenon in myocardium.

Anesthetics↗

[Development of apoptosis induced by whole-body irradiation in murine liver].

Apoptosis is known to be induced by radiation. However, the correlation between radiation-induced apoptosis and radiation injury in tumors in vivo has been unclear. In this paper, we report the study of apoptosis induced by whole-body irradiation using an immunohistochemical technique to detect DNA fragmentation in murine liver. A dose of 7 Gy was employed as LD50/30. DNA fragmentation was observed 30 min after radiation, and it peaked between 1 and 6 hours. DNA fragmentation could be detected 48 hours after radiation in capillary endothelium. This study was able to reveal the development of radiation-induced apoptosis in the liver by detecting DNA fragmentation in situ.

Animals↗

Immunohistochemical study of p53 expression in cancer tissues from patients undergoing radiation therapy.

Immunostaining using p53 monoclonal antibodies (p53(Ab-3) recognizes mutant type and p53(Ab-6) the wild type of p53 protein) was performed on frozen sections of biopsy specimens obtained before and during preoperative radiotherapy from 23 patients with head and neck squamous cell carcinoma. The positive staining rates of p53(Ab-3) before radiotherapy and at radiation doses of 4Gy, 10Gy and 20Gy were 30.0%, 38.9%, 25.0% and 6.25%, and those of p53(Ab-6) 10.5%, 11.8%, 5.0% and 0% respectively. The relationship between the immunohistochemical findings and antitumor effect at radiation dose of 20Gy was examined on the correspondent haematoxylin-eosin sections. In patients whose p53(Ab-3) stainings were positive at any doses of radiotherapy, the antitumor effect at the cumulative dose of 20Gy waas either remarkable or effective. Moreover, the frequency of the expression of mutant type p53 protein tended to increase in rather radiosensitive tumors. As for wild type p53 protein, there was no remarkable relationship between the staining of p53(Ab-6) and the antitumor effect.

Adult↗