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Biomedical subjects

Y Hara

Publications and source records attributed to Y Hara.

At least 361 records · Page 20Linked to original sources

Induction of anterior chamber-associated immune deviation with lymphoreticular allogeneic cells.

Immune privilege in the anterior chamber of the eye results in part from a selective deficit in delayed hypersensitivity that is elicited by antigenic materials placed in this unique tissue site. This stereo-typical systemic immune response (anterior chamber-associated immune deviation [ACAID]) to intraocular antigen can also be evoked in naive mice by the intravenous injection of syngeneic peritoneal exudate cells (PEC) that have been incubated overnight with a soluble antigen in the presence of supernatants of cultured iris and ciliary body (I/CB) cells or with transforming growth factor (TGF)-beta. To determine whether a similar protocol could be used to induce ACAID to transplantation antigens expressed on allogeneic PEC, we conducted experiments with allodisparate PEC donors and recipients selected to differ at loci dictating MHC and/or minor histocompatibility antigen. Recipients of I/CB supernatant-treated PEC from donors disparate for class I and/or class II antigens, as well as donors disparate only for minor H antigens, were specifically unable to display donor-specific delayed hypersensitivity, whereas recipients of PBS-treated allogeneic PEC developed intense donor-specific delayed hypersensitivity. We conclude that allogeneic PEC that have been cultured with I/CB supernatant overnight create an alloantigen-specific ACAID-inducing signal that confers ACAID on naive recipient mice.

Animals↗

Orthotopic liver retransplantation in rats.

A surgical experience with a method of rate orthotopic liver retransplantation (OLRT), and a preliminary study of immunological responses after OLRT are reported. OLRT was performed on the same recipient after the fist orthotopic liver transplantation (1st-OLT) according to our original (Kamada's) cuff method. Replacement of the portal vein (PV) and infra-hepatic vena cava (IHVC) cuffs was not technically difficult. However, there were no survivors from the first 6 retransplanted rats, mainly due to complications from defective supra-hepatic vena cava (SHVC) anastomoses. Unlike the human intra-abdominal SHVC, the posterior wall of the intra-abdominal SHVC in rats is too short and fragile to perform an end-to-end anastomosis twice between donor and recipient SHVC. For a second group of seven retransplants, a modification of the SHVC anastomosis was made between donor and recipient SHVC in conjunction with the recipient's cuff diaphragm. This enabled reanastomosis to be secure, resulting in the improved 1-week survival after isogenic OLRT (85.7%). This OLRT model has been applied to the fully allogeneic combination for several immunological studies and led to novel findings. Thus, an experimental model of a rat orthotopic liver retransplant model has the potential to allow more valuable insights into the immunological study of chronic rejection, sensitization and chimerism following liver retransplantation.

Anastomosis, Surgical↗

Interaction of class III antiarrhythmic drugs with muscarinic M2 and M3 receptors: radioligand binding and functional studies.

We have recently reported that class III antiarrhythmic drugs inhibit the muscarinic acetylcholine (ACh) receptor-operated K+ current (IK,ACh) in guinea-pig atrial cells by different molecular mechanisms. The data obtained from the patch-clamp study suggest that D,L-sotalol inhibits IK,ACh by blocking the muscarinic receptors, whereas MS-551 inhibits the K+ current by blocking the muscarinic receptors and depressing the function of the K+ channel itself and/or the guanine nucleotide-binding protein (G protein). This study was undertaken to determine whether the class III antiarrhythmic drugs D,L-sotalol and MS-551 interact with the muscarinic receptors of cardiac and peripheral tissues. Both drugs inhibited concentration dependently the specific [3H]N-methylscopolamine ([3H]-NMS) binding to membrane preparations obtained from guinea-pig atria and submandibular glands. The competition curves of these drugs for [3H]-NMS binding to glandular membranes were monophasic, suggesting competition with [3H]-NMS at a single site. Although the competition curve of D,L-sotalol for [3H]-NMS binding to atrial membranes was monophasic, that of MS-551 was biphasic and showed high- and low-affinity states of binding. D,L-Sotalol showed slightly, but significantly, higher affinity for cardiac-type muscarinic receptors (M2) than for glandular-type muscarinic receptors (M3). The inhibition constant (Ki) for MS-551 in glandular membranes was also slightly greater than the high-affinity Ki value for the drug in atrial membranes. In guinea-pig left atria and ilea, D,L-sotalol shifted the concentration-response curves for the negative inotropic effect and the contracting effect of carbachol in a parallel manner. The slopes of Schild plot were not significantly different from unity, suggesting competitive antagonism, and the pA2 for D,L-sotalol in left atria was slightly greater than that in ilea. MS-551 also shifted the concentration response curve for the negative inotropic effect of carbachol in atrial preparations to a greater extent than that for the contracting effect in ileal preparations, although MS-551 failed to show a pure competitive antagonism. These results suggest that both D,L-sotalol and MS-551 interact with cardiac M2 and peripheral M3 receptors, and that at high concentrations they exert anticholinergic activity in cardiac and peripheral tissues.

Animals↗

Isolation, characterization and chromosomal mapping of an actin gene from the primitive red alga Cyanidioschyzon merolae.

Based on the results of cytological studies, it has been assumed that Cyanidioschyzon merolae does not contain actin genes. However, Southern hybridization of C. merolae cell-nuclear DNA with a yeast actin-gene probe has been suggested the presence of an actin gene in the C. merolae genome. In the present study, an actin gene was isolated from a C. merolae genomic library using a yeast actin-gene probe. The C. merolae actin gene has no intron. The predicted actin is composed of 377 amino acids and has an estimated molecular mass of 42 003 Da. Southern hybridization indicated that the C. merolae genome contains only one actin gene. This gene is transcribed at a size of 2.4 kb. When Southern hybridization was performed with C. merolae chromosomes separated by pulsed-field gel electrophoresis, a band appeared on unseparated chromosomes XI and XII. A phylogenetic tree based on known eucaryote actin-gene sequences revealed that C. merolae diverged after the division of Protozoa, but before the division of Fungi, Animalia and Chlorophyta.

Actins↗

Enhanced cellular proliferative activity and cell death in chronic thyroiditis and thyroid papillary carcinoma.

For the analysis of cellular proliferative activity and cell death in thyroid diseases, the Ki-67 labeling index, bcl-2 protein expression and cell death of follicular epithelia by immunohistochemistry and in situ DNA nick-end labeling methods were evaluated in normal thyroid tissues as well as in surgical specimens from cases of Hashimoto's disease (16 cases), focal lymphocytic thyroiditis (13 cases), Graves' disease (15 cases), follicular adenoma (20 cases) and papillary carcinoma (43 cases). Cellular proliferative activity and cell death were both enhanced in cases of thyroiditis, including Hashimoto's disease and focal lymphocytic thyroiditis. Thyroids from patients with follicular adenoma and papillary carcinoma also showed increased cellular proliferative activity and cell death. In addition, predominant high cellularity and partial loss of bcl-2 protein expression in papillary carcinoma suggested that the overgrowth and dedifferentiation were associated with malignancy.

Carcinoma, Papillary↗

Ganglioside characterization of a cell line displaying motor neuron-like phenotype: GM2 as a possible major ganglioside in motor neurons.

We have examined ganglioside compositions and the presence of sulfated glucuronyl glycolipids of immortalized motor neuron-like cell lines, neuroblastoma-spinal cord (NSC) hybrid cell lines established by fusing mouse neuroblastoma N18TG2 with motor neuron-enriched embryonic spinal cord cells. Among NSC cell lines, only NSC-34 aggregates acetylcholine receptors on co-cultured myotube and expresses a receptor for S-laminin, a neuromuscular junction specific basal lamina protein. GM2, which is only a minor ganglioside component of CNS, was the major component in NSC-34 occupying almost 75% of total gangliosides, whereas GD1a and GM3 were major species in the parental N18TG2, which had only 8.5% GM2. These results indicated that NSC lines have unique ganglioside pattern that is distinctive from other nervous tissues, and this pattern, especially that of NSC-34 cells, might reflect the characteristics of mouse spinal motor neuron gangliosides. Sulfated glucuronyl paragloboside was demonstrated to be present in N18TG2, however, it could not be detected in either of NSC cell lines. Even though the pathogenesis of amyotrophic lateral sclerosis remains unknown, autoimmunological participation has been suggested. Because high-titered antibody against GM2 has been observed in a patient with amyotrophic lateral sclerosis-like disease, GM2 which is possibly expressed on the surface of motor neurons might serve as a potential target antigen in this disorder.

Animals↗

Chemoprevention of 2-amino-1-methyl-6-phenylimidazo[4,5-b]-pyridine (PhIP)-induced mammary gland carcinogenesis by antioxidants in F344 female rats.

Chemopreventive effects of the antioxidants 1-O-hexyl-2,3,5- trimethylhydroquinone (HTHQ), 3-O-ethylascorbic acid (EAsA), 3-O-dodecylcarbomethylascorbic acid (DAsA), green tea catechins (GTC) and ellagic acid on 2-amino-1-methyl-6- phenylimidazo[4,5-b]pyridine (PhIP)-induced mammary carcinogenesis were examined in female F344 rats. Groups of 20-21 6-week-old rats were maintained on a powdered diet containing 0.02% PhIP alone, PhIP together with 0.5% HTHQ, 1% EAsA, 1% DAsA, 1% GTC or 0.1% ellagic acid, these antioxidants alone or basal diet alone without supplement for 52 weeks. The survival rates of PhIP plus antioxidant groups at the end of the experiment were higher than that of the PhIP alone group. Sequential observation of palpable mammary tumors demonstrated only one tumor by week 52 in the PhIP plus HTHQ group, whereas 40% of the rats receiving PhIP alone had tumors by this time point. The final incidence of mammary adenocarcinomas was significantly decreased in the PhIP plus HTHQ group (4.8%, P < 0.01) as compared to the PhIP alone value (40%). Although statistically not significant, incidences of adenocarcinomas in the other antioxidant-treated groups (23.8-28.6%) were also lower than in the PhIP alone group. Furthermore, the incidence of large intestinal tumors in the PhIP plus HTHQ group (0%) showed a tendency to decrease relative to the PhIP alone group (16.7%). These results indicate that antioxidants, particularly HTHQ, exert a potent chemopreventive action against PhIP-induced carcinogenesis.

Adenocarcinoma↗

Premedication with famotidine augments core hypothermia during general anesthesia.

BACKGROUND: Animal studies have provided considerable evidence to support a role of histamine in the central nervous system in thermoregulation, and premedication with a histamine H2 receptor antagonist before general anesthesia is used to reduce the risk of acid aspiration. The authors investigated whether premedication with famotidine had an effect on thermoregulation during general anesthesia. METHODS: In a randomized, placebo-controlled study, 30 ASA physical status 1 or 2 patients, scheduled for open abdominal surgery, were given either placebo or 40 mg oral famotidine 3 h before induction of anesthesia. Epidural buprenorphine (4 micrograms/kg) was injected, and anesthesia was maintained with 0.4-0.6% isoflurane and 66% nitrous oxide in oxygen. The tympanic membrane temperature was measured to assess core temperature, and forearm-fingertip and calf-toe skin-surface temperature gradients were used to assess peripheral vasoconstriction. Tympanic membrane temperature triggering initial vasoconstriction (a skin temperature gradient of 0 degree C) identified the vasoconstriction threshold. RESULTS: Tympanic membrane temperature during surgery in the patients premedicated with famotidine was significantly less than those in the patients without famotidine. Famotidine significantly reduced the thermoregulatory threshold for vasoconstriction in the leg (35.0 +/- 0.5 degree C, P < 0.05), compared to that in the placebo group (36.4 +/- 0.6 degree C) Once triggered, thermoregulatory vasoconstriction produced a core-temperature plateau and no further hypothermia was observed for the duration of the study. Neither mean arterial pressure nor heart rate were significantly different between the two groups. CONCLUSIONS: Premedication with famotidine augments intraoperative hypothermia. The mechanism appears to be inhibition of centrally mediated thermoregulatory control.

Abdomen↗

The effects of green tea catechins (Polyphenon) on DL-ethionine-induced acute pancreatitis.

The effects of green tea catechins (Polyphenon) on DL-ethionine-induced acute pancreatitis in rats were examined. The acute pancreatitis induced in this study was characterized by moderate inter- and intrastitial edema and patchy acinar cell necrosis. In rats induced with acute pancreatitis by an intraperitoneal injection of DL-ethionine, the wet weight of the pancreas (0.47 +/- 0.059 g/100 g body weight; p < 0.05), the serum amylase (10,432 +/- 996 IU/L; p < 0.001), and the tissue concentration of lipid peroxides (19.5 +/- 1.78 nmol/mg tissue DNA; p < 0.001) were significantly increased compared with values obtained in control rats (0.39 +/- 0.037 g/100 g body weight, 5,639 +/- 1,568 IU/L, and 10.7 +/- 1.04 nmol/mg tissue DNA, respectively) injected with isotonic saline. In contrast, in rats injected with DL-ethionine and supplied with a green tea catechin solution as a beverage instead of water during the experimental period, the tissue of pancreas was almost-correct, and the wet weight of the pancreas (0.39 +/- 0.054 g/100 g body weight; p < 0.05), the serum amylase (5,716 +/- 708 IU/L; p < 0.001), and the concentration of lipid peroxides in tissue (11.5 +/- 2.15 nmol/mg tissue DNA; p < 0.001) were significantly decreased compared with values obtained in rats injected with DL-ethionine and supplied with water as a beverage. These data suggest that green tea catechins may have a protective effect on the pathogenesis of experimental acute pancreatitis.

Acute Disease↗

The influence of aging on skin temperature and hemodynamic changes during spinal anesthesia.

We investigated the influence of aging on the relationship between arterial pressure and skin temperature as a simple and indirect indicator of cutaneous blood flow. Sole and palm skin temperatures, sublingual temperature, heart rate, mean arterial blood pressure (MAP), and the anesthetic level as determined by cold discrimination, were measured before and during minor surgery under spinal anesthesia in patients under 65 years (young group) and above 65 years (elderly group). The sole skin temperature (Tsole) started to increase in the young group whose anesthesia level reached above L1-L2, and approached the sublingual temperature in those whose anesthesia level reached above T8-T10 after spinal injection. There was, however, no relationship between the anesthesia level and Tsole in the young group. The change in Tsole was less in elderly patients than that in young patients with the same decrease in MAP. These findings suggest that a decrease in peripheral resistance may not be the main cause of hypotension during spinal anesthesia in elderly patients.

Adult↗

SD-3212, a new class I and IV antiarrhythmic drug: a potent inhibitor of the muscarinic acetylcholine-receptor-operated potassium current in guinea-pig atrial cells.

1. By use of patch-clamp techniques, the effects of SD-3212, a novel antiarrhythmic drug, on the calcium current (Ica), the sodium current (INa) and the muscarinic acetylcholine-receptor-operated potassium current (IK.ACh) were examined and compared with those of bepridil in guinea-pig single atrial cells. 2. SD-3212 inhibited ICa and INa in a concentration-dependent manner. The IC50 values of SD-3212 for inhibition of ICa and INa were 1.29 microM and 3.92 microM, respectively. The steady state inactivation curves of ICa and INa were shifted in the hyperpolarizing direction in the presence of 1 microM SD-3212. Similar inhibition of ICa and INa was also observed with bepridil. The IC50 values of bepridil for depression of ICa and INa were 1.55 microM and 4.43 microM, respectively. 3. The muscarinic acetylcholine-receptor-operated potassium current (IK.ACh) was activated by the extracellular application of 1 microM carbachol in the GTP-loaded cells or by the intracellular loading of GTP gamma S, a nonhydrolysable GTP analogue. SD-3212 potently inhibited the carbachol- and GTP gamma S-induced IK.ACh and the IC50 values were 0.38 microM and 0.20 microM, respectively. These IC50 values were very close and about 10 times lower than those for inhibiting ICa and INa. Bepridil also suppressed the carbachol- and GTP gamma S-induced IK.ACh with the IC50 values of 0.69 microM and 0.84 microM, respectively. 4. In guinea-pig atrial cells stimulated at 0.2 Hz, carbachol at a concentration of 1 microM markedly shortened action potential duration. Both SD-3212 (0.1-1 microM) and bepridil (1-10 microM) reversed the action potential shortening in a concentration-dependent manner. The antagonizing effect of SD-3212 on the carbachol-induced action potential shortening was more potent than that of bepridil. 5. These results suggest that SD-3212 inhibits IK.ACh by depressing the function of the potassium channel itself and/or associated GTP-binding proteins. SD-3212 is a unique antiarrhythmic drug, which potently inhibits IK.Ach in addition to its class I and IV effects. SD-3212 and bepridil may be useful for the termination and prevention of vagally-induced atrial flutter and fibrillation.

Action Potentials↗

Histological study of lectin binding in regenerated rat junctional epithelium.

We investigated the expression of carbohydrate residues in regenerated junctional epithelium (JE) cells histopathologically with lectin staining to clarify the mechanisms responsible for the changes in their expression in JE cells derived from residual oral epithelium. Curettage and root planing procedures were performed on the buccal gingival sulci of rat first lower molars, and JE and connective tissues were completely removed. The mandibles were resected after 12 h and 1, 2, 3, 4, 5, 7, 10 and 14 days, fixed with paraformaldehyde, decalcified with EDTA and embedded in paraffin. Serial sections were stained histochemically with four kinds of lectins (PNA, DBA, GS I, UEA I) to clarify the expression patterns of carbohydrate residues in regenerating epithelium. No binding of PNA or DBA was observed even when the regenerating epithelium was attached to the root surface, and they showed the same negative reactions as the basal cells of oral gingival epithelium (OGE). Positive reactions were, however, observed on the more stratified regenerating epithelium along the root surface. Positive reactions with GS I and negative reactions with UEA I were observed throughout the regeneration process, and these were the same as those observed in the basal cells of OGE. Therefore, we concluded that the basal cells and regenerated epithelium derived from OGE expressed the same carbohydrate residues. Furthermore, the expression of carbohydrate residues, one of the characteristics of JE, was related not only to the attachment to the tooth surface but also to changes of cell shape and cytoskeleton with stratification along the root surface.

Animals↗

[Epidemiological study of tsutsugamushi disease in Gunma prefecture. A special field study and serotype].

An epidemiologically investigated of invasion of Rickettsia tsutsugamushi, inhabitant of mites and serum sample from patients with Tsutsugamushi disease in Gunma prefecture from 1984 to 1994 was made. Our data clearly indicated that Rickettsia tsutsugamushi was not located but widely spreaded throughout the Prefecture. Mites on rodents, were classified into 4 genus and 12 species and about 15% of them were Leptotrombidium pallidum and Leptotrombidium scuttellare, well known virulent vectors. The highest incidence rate of this disease was observed in the northwest area of the Prefecture from October to December, while a smaller number of patients occurred in other areas and in other months. About fifty percent of the serum samples from the patients were positive to the Karp strain. These results suggest that the major cause of this disease is the Karp strain and the disease could occur potentially in various areas of the Prefecture.

Animals↗

[Appearance of antibacterial activity of oxacillin against methicillin resistant Staphylococcus aureus (MRSA) in the presence of catechin].

We previously reported that tea catechin shows bactericidal activity against various bacteria including methicillin resistant Staphylococcus aureus (MRSA) and that bactericidal catechin damages the lipid bilayer of bacterial cell membranes. Here we describe that oxacillin (MPIPC) shows antibacterial activity against MRSA in the presence of catechin below MIC. Twenty clinical isolates of MRSA were examined by a cup method. In the absence of catechin, MPIPC even at a concentration of 40 micrograms/ml did not show antibacterial activity against all isolates of MRSA. However, when catechin below MIC (25-100 micrograms/ml) was mixed with the agar medium, MPIPC (5-12.5 micrograms/ml) showed antibacterial activity against all MRSA isolates. By counting the numbers of viable bacteria in a broth culture, only MPIPC (5 micrograms/ml) or catechin (100 micrograms/ml) showed similar growth curves to the control. But addition of both MPIPC and catechin reduced the number of viable bacteria to 1/100-1/10000 after 24 hours of cultures. Besides MPIPC, in the presence of catechin below MIC methicillin (12.5 micrograms/ml), aminobenzylpenicillin (32 micrograms/ml), tetracycline (2.5 micrograms/ml), and chloramphenicol (12.5 micrograms/ml) showed antibacterial activities against multiple drug resistant MRSA to antibiotics mentioned above. These findings suggest a possible use of catechin in the treatment of MRSA infection.

Catechin↗

Effects of various natural antioxidants on the Cu(2+)-mediated oxidative modification of low density lipoprotein.

We have reported in our previous paper that several flavan-3-ol derivatives (tea polyphenols) inhibited the Cu(2+)-mediated low density lipoprotein (LDL) oxidation in vitro. (-)-Epigallocatechin gallate (EGCG), in particular, exhibited strong inhibition. In this study, we have compared the antioxidative effects of EGCG with those of other natural antioxidants, such as flavonols, sesaminol, curcuminoid derivatives, tocopherol analogues and theaflavins. The antioxidative effects were monitored by conjugated diene formation in LDL which was carried out at 37 degrees C with 5 microM CuSO4 with or without antioxidants. Dibutyl hydroxytoluene (BHT) was used as a reference compound. The lag-time before the onset of conjugated diene formation was more than 100 min in the presence of 0.5 microM EGCG, theaflavin, myricetin, quercetin, and sesaminol. The ability to prolong the lag-time was in the order of sesaminol > quercetin > EGCG > theaflavin > or = myricetin > BHT > alpha-tocopherol. Among the 4 tocopherol analogues used, alpha-tocopherol showed the strongest antioxidative activity. We have also studied the effects of EGCG, BHT, and alpha-tocopherol on cholesteryl and alpha-tocopherol on cholesteryl ester (CE) degradation and apolipoprotein B 100 (apo B 100) fragmentation in the Cu(2+)-mediated oxidative modification of LDL. EGCG was the most effective inhibitor of CE degradation and apo B 100 fragmentation.

Antioxidants↗

Absorption of (-)-epigallocatechin gallate into rat portal vein.

Following oral administration of (-)-epigallocatechin gallate to rats, the presence of (-)-epigallocatechin gallate was examined in the portal blood. A compound present in the blood was identified as (-)-epigallocatechin gallate by HPLC and mass spectrometry analysis. The results clearly demonstrate that (-)-epigallocatechin gallate is absorbed, at least in part, into rat portal blood.

Absorption↗