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Biomedical subjects

S Yoshihara

Publications and source records attributed to S Yoshihara.

At least 91 records · Page 5Linked to original sources

Separation of outer cysts from metacercariae of Fasciola gigantica in an aqueous two-phase system.

The outer cysts (Oc) of Fasciola gigantica metacercariae (Mc) were separated in an aqueous two-phase system composed of polyethylene glycol and dextran in distilled water. A large number of separated Mc without Oc were recovered in the bottom layer rich in dextran in the system. Ten separated Mc in the layer were used to inoculate the mouse with a syringe and juveniles were recovered from the animals on the 14th day. When the number of juveniles recovered was compared with that from the mice infected orally with the same number of Mc, there was a significant difference between them (P < 0.05). The former method is easier than the latter for inoculation.

Animals↗

Inhibition of bronchoconstriction by pituitary adenylate cyclase activating polypeptide (PACAP 1-27) in guinea-pigs in vivo.

1. We studied the inhibitory effect of pituitary adenylate cyclase activating polypeptide (PACAP 1-27) on the increase in total pulmonary resistance (RL) caused either by allergen or histamine in anaesthetized, ventilated guinea-pigs. 2. PACAP 1-27 given via i.v. infusion (0.045-4.5 nmol kg-1 min-1) dose-dependently reduced the increase in RL caused by inhaled ovalbumin and histamine. At the highest dose, PACAP 1-27 prevented the increase in RL caused by ovalbumin and histamine completely. Infusion of PACAP 1-27 and the beta 2-adrenoceptor agonist, salbutamol (0.045-4.5 nmol kg-1 min-1) inhibited the increase in RL similarly, but salbutamol increased the heart rate more than PACAP 1-27. 3. PACAP 1-27 and salbutamol given via inhaled aerosol (0.1 mM, 20 breaths) significantly reduced the increase in RL caused by histamine infused i.v., whereas aerosolised sterile saline did not. Both PACAP 1-27 and salbutamol caused bronchodilator effects within 1 min of drug inhalation and these effects remained throughout the 20 min of study. 4. Because PACAP 1-27 produced significant bronchodilatation and rapid onset of sustained action in vivo and without pronounced cardiovascular side effects, we conclude that this peptide may have therapeutic potential as a bronchodilator.

Adenylyl Cyclases↗

Plasma extravasation in the rat trachea induced by cold air is mediated by tachykinin release from sensory nerves.

Cold air was delivered to anesthetized, artificially ventilated, pathogen-free F344 rats via a tracheal cannula. Inhalation of cold air increased Evans blue dye extravasation in the trachea in a time-dependent (1 to 10 min) manner. Plasma extravasation increased after 3 min exposure to cold air and reached a maximum after 10 min exposure. The neutral endopeptidase inhibitor, phosphoramidon (2.5 mg/kg, intravenously), increased by 84% the plasma extravasation induced by inhalation of cold air for 1 min. The plasma extravasation evoked by 5 min exposure to cold air was abolished by the NK1 tachykinin receptor antagonist, CP-99,994 (4 mg/kg, intravenously); was reduced 30% by the B2 bradykinin receptor antagonist, HOE140 (0.1 mumol/kg, intravenously); and was not affected by H1 (pyrilamine, 10 mg/kg, intraperitoneally) or H2 (cimetidine, 10 mg/kg, intraperitoneally) histamine receptor antagonists or the cyclooxygenase inhibitor indomethacin (5 mg/kg, intravenously). In rats infected with Sendai virus, plasma extravasation evoked by inhalation of cold air was greater than in pathogen-free rats. Pretreatment with CP-99,994 (4 mg/kg, intravenously) inhibited completely the plasma extravasation induced by cold air in virus-infected rats. These findings indicate that cold air increases plasma extravasation in the rat trachea by a neurogenic mechanism that involves the release of tachykinins from sensory nerves. Kinin release may also play a role in this neurogenic inflammatory response.

Air↗

Metabolism of 4-aminobiphenyl and 4-acetylaminobiphenyl in perfused guinea pig liver.

To obtain further knowledge of the metabolic fate of carcinogenic 4-aminobiphenyl (ABP) and 4-acetyl-aminobiphenyl (AABP) in intact guinea pig liver, we performed in situ liver perfusion using a recirculation method. Following a biexponential disappearance of ABP from the perfusate, not only AABP but also its secondary metabolites 4'-hydroxy AABP (4'-OH AABP) and 4-glycolylaminobiphenyl (GABP) appeared as the major metabolites. 4'-Hydroxy ABP (4'-OH ABP) was also detected in the perfusate as another predominant metabolite, while N-hydroxy ABP (N-OH ABP) as a minor one. Most of these metabolites, except for AABP and GABP, found in both perfusate and bile were conjugated with glucuronic acid. In addition, considerable amounts of all the metabolites were also detected in either unconjugated or conjugated form in a homogenate of the liver tissue after perfusion. When AABP was infused, an almost similar metabolic profile to that of ABP was obtained, but the amount of ABP was quite small and N-OH AABP rather than N-OH ABP was found. These results demonstrate that both ABP and AABP are rapidly metabolized by a combination of N-acetylation, C- and N-hydroxylations as well as glucuronidation in guinea pig liver.

Aminobiphenyl Compounds↗

[Brain CT studies in 26 cases of aged patients with Down syndrome].

Computed tomographic images of brains from 26 individuals (10 males and 16 females) with Down syndrome were analysed for roentogenographic measurement. Their ages ranged from 14 to 47 years, the average being 28 years. The results showed that their Sylvian fissure ratio was larger in the aged group. A high incidence of calcification in basal ganglia, choroid plexus and pineal body was noted (85%). An increased Sylvian fissure ratio and a high incidence of intracranial calcification may be practically used as representatives of premature aging. Furthermore, a high incidence of mega cisterna magna implicates that it is worthy of study whether individuals with Down syndrome have a predisposition to underdevelopment of cerebellum.

Adolescent↗

Analysis of schizont antigen of Leucocytozoon caulleryi by SDS-PAGE and western blot.

To investigate the molecular weight of the schizont antigen of Leucocytozoon caulleryi, the schizont antigen was analysed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and western blot. Thirty to 35 protein bands were detected by SDS-PAGE under reduced condition and ranged from 25 to 300 kDa. Most of them reacted with the sera of infected chickens by Western blot. The bands corresponding to polypeptides of 44, 58, 67 and 180 kDa reacted strongly, however, the bands of 30, 50 and 78 kDa did not react.

Animals↗

Relationship between hemolytic activity and adsorption capacity of aluminum hydroxide and calcium phosphate as immunological adjuvants for biologicals.

Aluminum hydroxide (Al) and calcium phosphate (Ca) gels have been used as vaccine adjuvants for many years. We investigated mechanism of the hemolytic activities of both adjuvant materials. The hemolytic activity of each gel depended on the gel dose. The adsorption capacities and the hemolytic activities of both adjuvants decreased as the concentration of phosphate increased in a gel-washing solution. A positive correlation between the hemolytic activity and the adsorption capacity was found in Al-gel. A disruptive effect of Ca-gel on membrane of erythrocytes was shown by electron microscopy. Ca-gel required more than 10 times as much pre-adsorbed ovalbumin as did Al-gel to inhibit the hemolysis. These results suggest that the hemolytic activity of both adjuvant materials depended mainly on the adsorption ability, and it may be useful to control the adsorption ability of adjuvants to reduce their hemolytic activity.

Adjuvants, Immunologic↗

Role of kinins in anaphylactic-induced bronchoconstriction mediated by tachykinins in guinea-pigs.

1. In the present study, we have investigated the role of kinins in allergen-induced bronchoconstriction. 2. Anaesthetized guinea-pigs were sensitized to ovalbumin, ventilated artificially, pretreated with atropine (1.4 mumol kg-1, i.v.) and total pulmonary resistance (RL) measured. In preliminary studies in the presence of the neutral endopeptidase inhibitor, phosphoramidon (4.5 mumol kg-1, i.v.), the bradykinin B2 receptor antagonist Hoe 140 (0.1 mumol kg-1, i.v.) completely abolished the increase in RL following aerosolized bradykinin (1 mM, 40 breaths), but had no effect on the increase in RL following aerosolized neurokinin A (NKA, 10 microM, 40 breaths). On the other hand, a combination of the NK1 (CP-96,345, 2 mumol kg-1, i.v.) and NK2 (SR 48968, 0.3 mumol kg-1, i.v.) tachykinin receptor antagonists abolished completely the increase in RL produced by NKA and partially inhibited the increase in RL produced by bradykinin. These results confirm previous studies that suggest that bradykinin induces the release of tachykinins from sensory nerves in guinea-pig airways. 3. Aerosolized ovalbumin (0.5%, 5 breaths) increased RL in sensitized guinea-pigs pretreated with atropine (1.4 mmol kg-1, i.v.), an effect that began within 2 min and reached a maximum within 5 min; RL remained above baseline at 20 min. Pretreatment with the bradykinin B2 receptor antagonist, Hoe 140, decreased the bronchoconstrictor effect of ovalbumin markedly at 10 to 20 min. In the presence of phosphoramidon (4.5 mumol kg-1, i.v.) the inhibition induced by Hoe 140 was apparent earlier and remained over the 20 min period of study. 4. Pretreatment with a combination of NK1 (CP-96,345) and NK2 (SR 48968) tachykinin receptor antagonists also markedly inhibited ovalbumin-induced bronchoconstriction; addition of the bradykinin B2 receptor antagonist to the NK1 and NK2 tachykinin receptor antagonists had no additional inhibitory effect on antigen-induced bronchoconstriction.5. These findings confirm that activation of sensory nerves to release tachykinins in guinea-pig airways contribute to antigen-induced bronchoconstriction, and provide evidence that tachykinin release is due to kinins generated during the allergic response.

Adrenergic beta-Antagonists↗

Evidence for reduction of bradykinin-induced bronchoconstriction in guinea-pigs by release of nitric oxide.

1. In this study the influence of nitric oxide (NO) on the bronchoconstriction induced by bradykinin in anaesthetized and artifically ventilated guinea-pigs pretreated with atropine was investigated. 2. Aerosol administration of bradykinin (0.1-1 mM, 40 breaths) caused a dose-dependent increase in lung resistance (RL): maximum increase in RL was 2.5 fold the baseline value. Pretreatment with aerosolized NG-nitro-L-arginine methyl ester (L-NAME) or NG-monomethyl-L-arginine (L-NMMA) (1 mM, 10 breaths every 5 min for 30 min), NO synthase inhibitors, markedly increased the bronchoconstrictor response to bradykinin. L-Arginine, but not D-arginine, (3 mM, 10 breaths every 5 min for 30 min) reversed the hyperresponsiveness to aerosolized bradykinin caused by L-NAME and L-NMMA. 3. L-NAME (1 mM, 10 breaths every 5 min for 30 min) increased the bronchoconstriction induced by intravenous bradykinin (1-10 nmol kg-1). L-Arginine, but not D-arginine, (10 breaths every 5 min for 30 min) reversed the hyperresponsiveness to intravenous bradykinin caused by L-NAME. 4. The increase in RL induced by capsaicin, either aerosol (10 microM, 10 breaths) or i.v. (20 nmol kg-1) was not affected by L-NAME (1 mM, 10 breaths every 5 min for 30 min). Acute resection of the vagi did not affect the bronchoconstriction evoked by bradykinin in guinea-pigs, either in the absence or presence of L-NAME (1 mM, 10 breaths every 5 min for 30 min). 4. These results suggest that, irrespective of the route of administration, bradykinin releases NO or a related molecule which exerts a bronchodilator action that opposes the bronchoconstrictor mechanisms activated by bradykinin itself.

Aerosols↗

Age-related changes in evoked otoacoustic emission in normal-hearing ears.

The evoked otoacoustic emissions (e-OAEs) elicited by both tone-bursts and clicks were investigated in normal-hearing ears of three age groups: young age group (approximately 30 years), middle age group (31-50 years) and old age group (51 years approximately). The pseudothreshold of tone-burst e-OAE at stimulus frequencies between 500 Hz and 2 kHz was significantly elevated in the middle and old age groups compared with that in the young age group. When the relationship between averaged emission cochleograms drawn from the mean pseudothresholds of tone-burst e-OAEs at 6 stimulus frequencies between 500 Hz and 4 kHz and averaged pure-tone audiograms was analyzed, a clear difference was found among the three age groups in a frequency range between 500 Hz and 2 kHz. Total echo power, reproducibility, highest peak power and frequency area peak power up to 4 kHz in click e-OAE significantly decreased in the middle and old age groups compared with those in the young age group. These results indicate the possibility that the function of cochlear micromechanics deteriorates in all cochlear partitions with increasing age, even in normal-hearing ears.

Acoustic Stimulation↗

Pathophysiology of hearing impairment in acoustic neuroma with profound deafness: analysis by evoked otoacoustic emission and promontory stimulation test.

Evoked otoacoustic emissions (e-OAEs) were examined and promontory stimulus test (PST) done prior to tumor removal in 15 cases of surgically proven unilateral acoustic neuroma (AN) with more than 80 dB in pure-tone hearing level. The e-OAEs were elicited by tone-bursts at 1 kHz and 2 kHz, and an interaural difference in pseudothreshold of more than 10 dB was defined as a significant impairment of the cochlear function. In PST, electrical stimulation was applied by a burst mode at 6 stimulus frequencies between 50 Hz and 1600 Hz. The number of stimulus frequencies with a positive response was defined as the PST score. The results obtained were as follows; i) Mean interaural difference in e-OAE pseudothreshold was 16.0 dB at 1 kHz and 8.0 dB at 2 kHz, ii) Positive PST was found in only 40% of the subjects and the mean PST score was 0.93, iii). When the pathophysiology of hearing impairment was inferred from the combination of the findings in e-OAEs and PST, it was found that 33.3% of the subjects had pure cochlear impairment, 13.3% pure retrocochlear impairment and 46.7% cochlear-retrocochlear impairment. It is concluded that analysis of both e-OAEs and PST could clarify the pathophysiology of hearing impairment in AN with profound deafness, which cannot be determined by conventional psychoacoustic tests.

Acoustic Stimulation↗

Partial characterization of larval antigens of Strongyloides papillosus by western blot analysis.

Western blotting was used to analyse the antigens in extracts of adult and infective larvae (SIL) of Strongyloides papillosus with serum of infected rabbits. Several proteins in both the extracts reacted with immunoglobulin G in the serum. A protein of about 16 kDa in the extract of SIL showed an especially strong reaction. No cross-reaction was observed among proteins in the extracts of adult worms of such cattle helminths as Fasciola gigantica, Homalogaster paloniae, Trichuris discolor and Setaria digitata to S. papillosus-infected rabbit serum. The cuticle of the worms and substances in the worm tracks showed positive reaction with the immunoperoxidase test on SIL with the serum.

Animals↗

[Chromogranin A like immunoreactivity in respiratory tissues of ovalbumin-sensitized guinea pigs].

Chromogranin A like immunoreactivity (CGA-IR) was measured by a specific radioimmunoassay using N-terminal specific antiserum R-0763 in respiratory tissues of guinea pigs. All guinea pig were actively immunized by recurrent Ovalbumin (OA)-inhalation and divided into two groups; one was challenged by OA, and the other inhaled saline as a control. These groups were studied for respiratory resistance and sacrificed for measurement of CGA-IR concentrations in the trachea, major bronchus and lower lung. In the control group, CGA-IR level was 0.4-2 pmol/g wet weight of tissue, and its distribution order was lower lung > trachea > major bronchus. In the OA-challenged group, provocation clearly induced significant elevation of CGA-IR in the trachea and main bronchus coinciding with elevation of respiratory resistance. In the lower lung, on the other hand, the increase in CGA-IR was not significant. These results suggest that experimental immediate asthmatic response (IAR) possibly acts as a stressor to the sympathetic nervous systems in guinea pig air-ways.

Airway Resistance↗

A study of the bioactive bone cement--bone interface: quantitative and histological evaluation.

The interface between bone and a bioactive glass cement--a mixture of bioactive glass powder and ammonium phosphate solution, previously reported on by the authors--was evaluated quantitatively and histologically. The materials tested were (1) the original bioactive glass cement (BCI cement); (2) an improved type of bioactive glass cement (BCII cement); (3) polymethylmethacrylate (PMMA) bone cement; and (4) a bioactive, apatite-wollastonite-containing, glass ceramic (A-WGC). Hardened cylindrical specimens of each cement were inserted loosely into canine femora and the interfacial shear strengths were measured using a push-out test. The interfacial strength values of the bioactive glass cements increased with prolonged implantation time. At each postimplantation time studied (8, 12, and 24 weeks), the interfacial strength value of BCI cement did not differ significantly from that of A-WGC. BCII cement interfacial strength was greater than that of BCI cement, whereas the interfacial strength of PMMA bone cement remained at a very low level throughout the study. Histological examinations revealed that direct bonding of both bioactive glass cements to bone had occurred without pathologic degradation. After 24 weeks, the defects between the bone and the bioactive glass cements had been filled with mature lamellar bone. Because the bioactive glass cement system developed by the authors, especially BCII cement, shows excellent osteoconductivity and bonds to bone tightly, we consider it to be a promising material for fixing prostheses into bone.

Animals↗

A new bioactive bone cement consisting of BIS-GMA resin and bioactive glass powder.

We have developed a bioactive bone cement consisting of silane-treated CaO-SiO 2-P 2O 5-CaF 2 glass powder as the filling particles and bisphenol-a-glycidyl methacrylate (BIS-GMA) diluted with triethylene-glycol dimethacrylate (TEGDMA) as the organic matrix. Histological examination demonstrated direct bonding between the cement and bone along the circumference of the cement at 4 weeks after implantation in rat tibia. The compressive strength and toughness of the cement were two and four times greater than those of polymethylmethacrylate (PMMA) cement, respectively. The inflammatory reaction of the skin caused by the new cement was not as intense as that for PMMA 3 days after subcutaneous implantation. This new cement may be applicable as a bioactive bone cement with high mechanical strength.

Animals↗

Enzymic determination of acidic glycoconjugates in human pancreatic juice.

Acidic glycoconjugates (glycosaminoglycans, sulfated glycopeptide, and sialoglycopeptide) were isolated by precipitation with cetylpyridinium chloride from human pancreatic juice after digestion with pronase. The acidic glycoconjugates were found exclusively in the proteinaceous precipitate that occurred during dialysis against a buffer of low ionic strength. The concentration of the acidic glycoconjugates in normal pancreatic juice was about 2.4 mg/L. The acidic glycoconjugates were characterized by electrophoresis on cellulose acetate membrane and chemical analysis before and after digestion with Streptomyces hyaluronidase, chondroitinase AC, chondroitinase ABC, and heparitinase. It was found that the major acidic glycoconjugates were heparan sulfate (39.3%), sulfated glycopeptide (34.4%), chondroitin sulfate (14.2%), and the minor ones hyaluronic acid (6.4%) and sialoglycopeptide (5.7%).

Chondroitinases and Chondroitin Lyases↗