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Biomedical subjects

T Fujinaga

Publications and source records attributed to T Fujinaga.

At least 19 recordsLinked to original sources

Physiological production of singlet molecular oxygen in the myeloperoxidase-H2O2-chloride system.

The putative role of singlet oxygen (1O2) in the respiratory burst of neutrophils has remained elusive due to the lack of reliable means to study its quantitative production. To measure 1O2 directly from biological or chemical reactions in the near infrared region, we have developed a highly sensitive detection system which employs two InGaAs/InP pin photodiodes incorporated with a dual charge integrating amplifier circuit. Using this detection system, we detected light emission derived from a myeloperoxidase (MPO)-mediated reaction in physiological conditions: pH 7.4, 1-30 nM MPO, 10-100 microM H2O2 and 100-130 mM CI in place of Br without the use of deuterium oxide. The MNPO-H2O2-CI(-) system exhibited a single emission peak at 1.27 microm with a spectral distribution identical to that of delta singlet oxygen. Our results suggest physiological production of 1O2 in the MPO-H2O2-CI(-) system at an intravacuolar neutral pH. The MPO-mediated generation of 1O2, which may have an important role in host defense mechanisms, is discussed in connection with previous results.

Animals

Surgical correction of temporomandibular joint ankylosis in two cats.

Two cats with temporomandibular joint ankylosis were examined. Difficulty in opening the mouth and malocclusion were associated with trauma and fracture around the zygomatic arch and the condyle of the mandible after an automobile accident. Unilateral condylectomy or aggressive caudal mandibulectomy was performed to improve quality of life. Excision arthroplasty of the temporomandibular joint improved mandibular function and appearance. Some malocclusion remained but had no clinical significance.

Accidents, Traffic

Peri-operative changes in echocardiographic measurements and plasma atrial and brain natriuretic peptide concentrations in 3 dogs with patent ductus arteriosus.

Peri-operative changes in echocardiographic measurements and plasma levels of atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) were investigated for 1 month in 3 dogs with patent ductus arteriosus (PDA). Post-operative left ventricular end-diastolic dimention and fractional shortening decreased in all cases. Pre-operatively increased plasma ANP concentrations reduced dramatically after the operation. Peri-operative changes in plasma BNP levels had slightly S-shaped curves in all cases. These observations suggest that post-operative responsiveness of ANP and cardiac function are rapid in comparison with cardiac morphological changes, and BNP has a different pathophysiological significance from ANP in dogs with PDA.

Animals

Endothelial cell responses to chitin and its derivatives.

The effects of chitin and its derivatives on the proliferation of human umbilical vein endothelial cells (HUVECs) and on the production of cytokines were examined in vitro. Chitin and its derivatives had no effect on the proliferation of cultured HUVECs. N-Sulfonated 70% deacetylated chitin (S-DAC70) stimulated the production of interleukin (IL)-1beta, IL-6, IL-8, and tumor necrosis factor (TNF)-alpha from HUVECs. Compared to S-DAC70, the other materials tested in the present study showed less effect in the stimulation of IL-8 and TNF-alpha production and had no effect in the stimulation of IL-1beta and IL-6 production. These results indicated that S-DAC70 affects HUVECs function but not proliferation.

Biocompatible Materials

Relationship between glutathione S-transferase M1 deficiency and urothelial cancer in dye workers exposed to aromatic amines.

PURPOSE: It is speculated that the susceptibility to urothelial cancer in dye workers who are exposed to aromatic amines is affected not only by occupational environmental factors but by host specific factors. We evaluated the interaction between glutathione S-transferase M1 gene deficiency and the occupational environmental factors associated with urothelial cancer. MATERIALS AND METHODS: The study included 137 workers who had prior exposure to dyestuff intermediates, of whom 36 had urothelial cancer. The prevalence of a glutathione S-transferase M1 gene polymorphism was investigated using polymerase chain reaction. The relationship between the glutathione S-transferase M1 0/0 gene and occupational environmental factors in the onset of urothelial cancer was examined by multivariate analysis. RESULTS: The prevalence of glutathione S-transferase M1 gene deficiency did not differ significantly between the urothelial cancer (21 cases, 58.3%) group and the cancer-free (47, 46.3%) group. It was estimated that 29.6% of the urothelial cancers in these dye workers was attributable to the glutathione S-transferase M1 0/0 gene. Analysis using multiple logistic models showed low predictive ability for urothelial cancer due to glutathione S-transferase M1 gene deficiency (p = 0.084, odds ratio 2.260, 95% confidence interval [CI] 0.904 to 5.652). A history of working in small factories (p = 0.000, odds ratio 7.404, 95% CI 2.854 to 19.206) and a long period of exposure (p = 0.016, odds ratio 5.051, 95% CI 1.371 to 18.612) significantly predicted cancer. CONCLUSIONS: We demonstrated a strong trend using the multiple logistic analysis of the contribution of glutathione S-transferase M1 gene polymorphism and occupational environmental factors. Therefore, the glutathione S-transferase M1 enzyme might have an important role in the detoxification of aromatic amine derived carcinogens. Occupational environmental factors, however, might contribute more than a glutathione S-transferase M1 gene deficiency to the occurrence of urothelial cancer among individuals exposed to aromatic amines, because of the extremely potent carcinogenicity of some occupational environmental factors.

2-Naphthylamine

Measurement of serum and synovial fluid keratan sulphate and antibody to collagen type II in equine osteoarthritis.

Keratan sulphate (KS) concentration and anticollagen type II antibody levels were measured in the serum and synovial fluid (SF) of clinically normal horses and horses with osteoarthritis (OA). Serum KS in OA was significantly higher than that in normal horses, while no significant difference was found in KS levels of SF between normal and OA. Assays of antibody to collagen type II showed no significant increase in sera and SF of OA. It was suggested that levels of serum KS would be of value in the pathological detection of OA in the joint, although there was no evidence that the measurable autoimmunity to collagen antigens would reflect the process of OA.

Animals

Spectrophotometric determination of neutrophil cytochrome b558 of chronic granulomatous disease.

BACKGROUND: Chronic granulomatous disease (CGD) is an inherited disease characterized clinically by severe recurrent bacterial infections from infancy. This disease is a disorder of the formation of superoxide (O2-) by the neutrophil NADPH oxidase system, mostly due to defects in cytochrome b558 (cyt b558), which is one of the oxidase components. Diagnosis of CGD has been performed by the assay of the O2- forming activity, immunological determination of defects in the oxidase components, and or spectrophotometry of cyt b558. However, spectrophotometric analysis of the b-type heme is difficult with small amounts of blood from infant CGD patients, as the limited amounts of neutrophils are contaminated with a relatively high ratio of hemoglobin (Hb) that interferes with the heme spectrum of cyt b558. This report presents an accurate method for the spectrophotometric analysis of cyt b558 in a small amount of CGD neutrophils that were treated with CO gas in a safe procedure instead of the previously reported CO-bubbling method. METHODS AND RESULTS: The difference of the reduced minus oxidized cyt b558 spectrum was measured under no interference from oxy Hb at the alpha and beta bands and differentiated as d[delta A]/d lambda (lambda = wavelength) to obtain further evidence for the defects of the cyt b558 heme spectrum. The interference from CO-insensitive met Hb was eliminated by subtracting the absorption peak at the Soret (gamma) band of the contaminating met Hb, which was estimated from the CO-treated and untreated spectra of the same, hemolyzed sample. CONCLUSIONS: This spectrophotometric method is feasible for the determination of abnormality and heme content of cyt b558 with a small amount of CGD neutrophils in 10-20 mL of blood even in the presence of contaminating Hb.

Cytochrome b Group

Apoptosis induction of POS canine osteosarcoma cells by vitamin D and retinoids.

Vitamin D3: 1-alpha, 25(OH)2D3 (calcitriol), 22-oxa-1,25(OH)2D3 (OCT), cholecalciferol (vitamin D3), and retinoids: all-trans retinoic acid (ATRA) and 9-cis retinoic acid, induced morphological changes in POS canine osteosarcoma cells into elongated, spindle or fibroblast like-shaped cells, and apoptotic like cell death characterized by cell shrinkage, condensation and margination of the nucleus for all drugs at 10(-6)M-10(-9)M after 72 to 120 hr culture. Apoptosis as shown by DNA laddering was induced at 48 hr by all drugs at 10(-6)M, 10(-7)M at 96 hr, 10(-8)M and 10(-9)M at 120 hr respectively. These vitamins are suggested to adjunct antineoplastic agents in canine osteosarcoma therapy by induction of apoptosis.

Animals

Ultrasonographic evaluation of portal vein hemodynamics in experimentally bile duct ligated dogs.

The purpose of this study was to evaluate the relationship between the results of laboratory examinations and ultrasonographic findings, especially portal vein hemodynamics in experimentally bile duct ligated dogs. Biliary obstruction was accomplished by surgically occluding the common bile duct in five dogs. All the dogs became visibly jaundiced within 24 hours after surgery. The total protein and albumin/globulin ratio showed a gradual decrease throughout the examination period, while blood urea nitrogen reached its peak in the 6th week and decreased to pre ligation values by the 10th week. Similar trends were noted in the alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, and direct and total bilirubin. Total cholesterol and fasting serum bile acid levels rapidly increased after surgery to peak values between the 2nd and 4th week, and then gradually decreased, but still remained high throughout the experiment period. The portal flow volume and velocity significantly (p < 0.05) decreased while only a slight increase was noted in the congestion index after bile duct ligation. The cross sectional area of the portal vein changed insignificantly. Bile duct and gallbladder distention was evident within the 1st week after ligation but there was little change in the echogenicity of the liver parenchyma. The results of this study suggest that the determination of Doppler ultrasound parameters of hepatic hemodynamics, especially the portal vein flow indices, may contribute to a better noninvasive assessment of the canine patient with biliary obstructive disease.

Animals

Serum copper and ceruloplasmin activity at the early growing stage in foals.

Serum concentrations of copper (Cu), zinc (Zn), manganese (Mn), calcium (Ca) and inorganic phosphorus (P), as well as antigenic ceruloplasmin (Cp) and oxidase activity as a functional index for copper metabolism, were measured in 10 foals (5 males and 5 females) and their dams. Samples were harvested from the foals within 1 wk after birth and monthly from 1 to 17 mo of age. Samples were collected from their dams in the perinatal period (monthly from 2 mo before delivery to 5 mo postpartum). Serum oxidase activity, antigenic Cp and Cu in foals were extremely low at 1 wk. Serum Cp had the lowest value of 17.0 +/- 8.0 (mean +/- SD) mg/dL within the 1st wk, then increased rapidly up to 43.7 +/- 5.8 mg/dL at 1 mo, and maintained this level until the 17th mo. Serum Zn in foals had the highest value of 73.2 +/- 13.1 micrograms/dL within 1 wk, then decreased to 38.3 +/- 5.9 micrograms/dL by 17 mo. Serum Mn, Ca and P in mares were almost stable and within established reference ranges for our laboratory in the perinatal period, and these values in foals were also in the normal range. Even on appropriate feeding, serum Cu, Cp and oxidase activity were quite low a few weeks after birth, while a higher proportion of Cp-binding copper was found in the foals. This might be caused by the limited synthesis of ceruloplasmin in this period. These data suggest that newborn foals are in a critical situation of marginal copper status in the early stage of growth.

Aging

Establishment of a monoclonal antibody (1/14/16H9) for detection of equine keratan sulfate.

OBJECTIVES: To establish a sensitive and specific monoclonal antibody (MAB) against equine keratan sulfate (KS) and to develop an enzyme immunoassay for measurement of the concentration of KS in serum and synovial fluid from horses. SAMPLE POPULATION: 18 synovial fluid and 48 serum samples were obtained from clinically normal horses and horses with arthritis. PROCEDURE: BALB/c mice were immunized with chondroitinase-ABC-digested proteoglycan monomer from equine joint cartilage, and MAB were raised, using Sp2/O cells as a fusion partner. A competitive ELISA was optimized, using one of the established MAB, and KS concentration in synovial fluid and sera from horses was measured. RESULTS: The high-titer MAB1/14/16H9, which specifically recognizes the epitope on equine KS, was identified. This antibody had no reactivity with chondroitin sulfate and core protein of proteoglycan monomers, hyaluronan, heparin, dermatan sulfate, and heparan sulfate. A competitive ELISA for determination of KS concentration was optimized, using this antibody. Precision data were obtained for the range of 10 to 160 ng/ml. The within- and between-assay coefficients of variation were 10.0 and 12.7%, respectively. CONCLUSIONS: MAB 1/14/16H9 that specifically recognized equine KS was established and was used to develop an enzyme immunoassay for measurement of the concentration of KS in synovial fluid and sera from horses. It is expected that the assay system using MAB 1/14/16H9 will contribute to evaluation of cartilage metabolic activity in horses.

Animals

Effects of chitin and its derivatives on the proliferation and cytokine production of fibroblasts in vitro.

The effects of chitin and its derivatives on the proliferation of fibroblasts and on the production of cytokines were examined in vitro. Chitin and its derivatives showed almost no acceleratory effect on the proliferation of cultured fibroblasts. On the contrary, high-concentration 500 micrograms ml-1) D-glucosamine cultures supplemented with or without a 10% fetal calf serum (FCS) supplementation showed a significant (P < 0.05) reduction in the rate of proliferation of L929 fibroblast cells relative to control. High-concentration chitosan cultures supplemented with 10% FCS showed a significant (P < 0.05) reduction in the rate of L929 fibroblast proliferation. However, the inhibition of cell proliferation by high concentrations of chitosan did not show in cultures without FCS. Interleukin-8 (IL-8) was induced in the supernatants of rat primary cultured dermal fibroblasts stimulated with chitin and its derivatives. Chitin and its derivatives did not stimulate the production of IL-6 by mouse dermal primary cultured fibroblasts. IL-1 alpha, IL-1 beta and tumour necrosis factor-alpha were not detected in the fibroblast supernatants. These observations support the notion that cell proliferation is accelerated indirectly by chitin and its derivatives when these materials are used in vivo. In vivo findings of a angiogenesis and migration of neutrophils may be due to persistent release of IL-8 from fibroblasts.

Animals

Consideration of the optimum pH for the analysis of serum p-phenylenediamine oxidase activity in thoroughbred horses.

The optimum pH for the measurement of serum p-phenylenediamine oxidase (Ox) activity was given (pH 6.6), and the relationship between serum ceruloplasmin (Cp) concentration and its Ox activity was established in healthy adult horses. In adult horses, serum antigenic Cp concentrations were measured by the single radial immunodiffusion (SRID) method with the affinity-purified antibody to equine plasma Cp and compared with its Ox activity. Efficient co-relation between Cp concentration and Ox activity in the sera (r = 0.93) and its Ox/Cp ratio were given. These results might contribute to the calculation of antigenic Cp concentration from its Ox activity, the analysis of holo-Cp content in serum, and the research for copper metabolism in thoroughbred horses.

Animals

Comparison of systemic and renal hemodynamics measured by Doppler ultrasonography in canine experimental hypovolemia.

The aim of this study was to examine renal hemodynamics at the hypovolemic and recovery phases in two different hypovolemic shock models using Doppler ultrasonography, and to compare this with systemic hemodynamics. In experiment 1, the hypovolemic phase was induced in 6 mongrel dogs by removing arterial blood at 30 ml/kg for 60 min. In the recovery phase, this blood was reinfused at 30 ml/kg over 60 min. In experiment 2, hypovolemia was induced in 12 beagle dogs by rapid blood removal until blood pressure decreased to 40 mmHg and was maintained at this pressure for 30 min. Six of the dogs were then infused with 20 ml/kg hydroxyethyl starch over 5 min, and the other 6 were infused with 60 ml/kg lactated Ringer's solution also over 5 min. Parameters for systemic and renal hemodynamics were measured by using a polygraph and the Doppler method, respectively. The decrease of diastolic blood flow, resulted in an increase of vessel resistance, and was detected in the hypovolemic kidney by the Doppler method. The rapid and large volume infusion of resuscitation fluids was effective for the recovery of both systemic circulation and renal blood flow, however this induced an increase of kidney vessel resistance, a result of the autoregulation mechanism of the kidney. The changes in these parameters at the main renal artery and interlobar artery were similar.

Animals

Evaluation of the catabolic activity of cartilage by measurement of serum keratan sulfate concentration in foals.

OBJECTIVES: To determine keratan sulfate (KS) concentration in the serum of foals at the early stage of growing, and to evaluate the role of serum KS as a cartilage catabolic marker, comparing its values with the fluctuation of serum alkaline phosphatase (ALP) activity as a measurement of osteoblastic activity. ANIMALS: 12 foals with normal growth and 3 foals with joint abnormalities within 18 months after birth. PROCEDURE: Measurement of KS concentration and ALP activity in serum and radiographic and physical examinations were done. RESULTS: In all foals, serum KS concentration was high from 1 week after birth to 3 months of age, while serum ALP decreased with aging. The value started to decrease rapidly from 3 to 5 months of age, then gradually reached adult values. During the first 3 months, KS concentration in male foals was significantly higher than that in female foals. In 3 foals which had joint problems, KS concentration was higher than that in normally growing foals at 1 week, and at 1, 2, and 3 months of age. CONCLUSIONS: Cartilage catabolic activity is higher in developing foals up to 3 months of age, suggesting that the immature joint at this time could be easily affected by any factor of loading. Moreover, though only 3 diseased foals were examined, higher serum KS concentration in these foals suggest that this variable might be a useful measure of joint diseases, even at an early stage of life in foals.

Aging

Comparison of regional myocardial technetium-99m-MIBI uptake between ECG-gated and ungated SPECT imaging.

UNLABELLED: Technetium-99m-MIBI uptake was compared between ECG-gated and ungated SPECT images in 10 normal subjects and 10 patients with coronary artery disease to investigate the effects of wall motion on regional myocardial uptake. METHODS: Left ventricular ejection fraction (LVEF) and wall motion were evaluated using the first-pass data acquired immediately after injection of 1110 MBq 99mTc-MIBI. A transaxial ungated image was reconstructed with cumulative data during a cardiac cycle. For transaxial gated images, data during a cardiac cycle were divided into eight frames and the first seven frames were used. The lateral/septal ratios for ungated and gated images were obtained using the counts in the ROIs drawn in the lateral and the septal walls. RESULTS: In 10 normal subjects, the lateral/septal ratio for gated increased during end-systole. The mean of the lateral/septal ratio was significantly lower for ungated than for gated (1.025 vs. 1.077, p < 0.05). In patients with septal wall asynergy and a normokinetic lateral wall, the mean of the lateral/septal ratios was significantly lower for ungated than for gated (1.267 vs. 1.325, p < 0.01). CONCLUSION: Ungated SPECT acquisition may underestimate regional myocardial uptake when myocardial wall motion is good, therefore, ECG-gated data should be acquired for accurate assessment of regional myocardial uptake of 99mTc-MIBI.

Adult

Ultrasonography, biochemical and hematological profiles in liver disease caused by intravenous administration of dimethylnitrosamine in dogs.

The results of liver function tests and ultrasonographical findings were analysed in 7 dogs that were intravenously injected dimethylnitrosamine (DMNA 2 mg/kg body weight) on 2 consecutive days each week for 10 weeks. Typical clinical signs and similar changes in liver enzyme concentrations that develop in dogs with natural cirrhosis were observed in this canine model. Severe anaemia and a significant reduction in the platelet numbers occurred in the dogs that died in the 5th week, while in all the other dogs these parameters decreased slightly. Serum total protein and the albumin/globulin ratio decreased gradually while the alkaline phosphatase, alanine amino transferase, aspartate amino transferase and gamma glutamyl transpeptidase activities increased significantly (p < 0.05) in all dogs after beginning the administration of DMNA. Ultrasound findings of a coarsened and heterogeneous echo pattern with increased echogenicity that are characteristic of canine cirrhosis were noticed at the same time when the changes in liver enzymes became evident. Present results suggest that ultrasonography in conjunction with liver function tests may be useful in the evaluation of experimentally induced liver cirrhosis.

Alanine Transaminase

[Urinary schistosomiasis: report of a case].

A 31-year-old Japanese man visited our department with macroscopic hematuria. He had stayed in Asia, Middle East and Africa during the past several years. He was diagnosed with urinary schistosomiasis because examination of urine and a biopsy specimen of the bladder mucosa revealed Schistosoma haematobium ova. Cystoscopy revealed granulomatous lesions and polypoid patches at the right ureteral orifice, posterior wall and dome of the bladder. Radiological examination showed no obstructive uropathy. Histologically, moderate to severe urothelial dysplasia was found but without evidence of malignancy. The disease has been under good control following treatment with praziquantel. Today international exchange between Japan and other countries is common. Therefore, physicians should consider the possibility of schistosomiasis in patients who have been to endemic areas.

Adult