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Biomedical subjects

K Harada

Publications and source records attributed to K Harada.

At least 775 records · Page 43Linked to original sources

[In situ hybridization histochemistry for the kidney diseases].

Crucial points of in situ hybridization methods were pointed out for study on the kidney diseases at molecular level. As examples of in situ hybridization study on kidney, we introduced two cases: 1) expression of the hepatocyte growth factor gene during renal regeneration after unilateral nephrectomy, and 2) induction of the metallothionein gene expression by ischemic acute renal failure. We proposed an idea to establish a probe bank for the in situ hybridization study.

Acute Kidney Injury↗

[A case report of a 63-year-old patient with malignant hyperthermia].

We have experienced a case of fulminant malignant hyperthermia who was a 63-year-old female weighing 44 kg. There was no particular past history nor family history. She underwent right mastoidectomy because of chronic otitis media. Her preoperative physical status was ASA I. She was premedicated with diazepam 10 mg and loxatigine 75 mg P.O. The induction was done with thiamylal 200 mg IV and fentanyl 0.1 mg IV followed by vecuronium 6 mg IV for endotracheal intubation. Intubation was easy and uneventful. Anesthesia was maintained with nitrous oxide 3 l.min-1, oxygen 3 l.min-1 and enflurane 2.0%. Seventy min after the induction of anesthesia, arterial blood gas analysis showed severe respiratory acidosis (PCO2: 63.2 mmHg, pH: 7.27) and it was improved with manual hyperventilation at that time. Pulse rate increased from 80 to 115 b.p.m. 20 minutes later. Then, the patient was ventilated with 100% oxygen, and anesthetic circuits and machine were exchanged for new units. Surgery was postponed. Muscle stiffness of upper extremities was observed and her temperature increased to a maximum of 38.9 degrees C. Surface cooling was started and dantrolene sodium 60 mg and furosemide 20 mg were given intravenously. The patient was transferred to the intensive care unit, and clinical signs improved gradually within one hour. Serum enzymes; CPK, LDH, GOT and GPT increased on the first postoperative day. On the 11 th postoperative day skeletal muscle biopsy was done under local anesthesia with 1% procaine and Ca-induced Ca-release rate test revealed positive for enflurane. This is the oldest patient of malignant hyperthermia reported in Japan.

Anesthesia, Inhalation↗

Human pulmonary hypoplasia. Statistical, morphological, morphometric, and biochemical study.

Human pulmonary hypoplasia was studied statistically and pathologically in a large series of autopsy cases. Multiple logistic regression analysis indicated five independent risk factors from 10 statistically significant factors for pulmonary hypoplasia: (1) hydrops fetalis; (2) renal anomalies; (3) hernia, including diaphragmatic hernia and omphalocele; (4) skeletal anomalies; and (5) abnormalities of amniotic fluid, such as oligohydramnios and polyhydramnios. The characteristics of pulmonary hypoplasia for each factor were defined by morphological, morphometric, and biochemical methods. All bronchiolar branching, acinar complexity, and acinar maturation were retarded in hypoplastic lungs with hydrops fetalis, renal anomalies, affected side of diaphragmatic hernia, omphalocele, and skeletal anomalies. Only acinar complexity and maturation were impaired in the lung with oligohydramnios due to prolonged rupture of membranes. The pathogenesis of pulmonary hypoplasias should be considered differently with each associated anomaly and time of impairment. While impairment in early gestational stage before 16 weeks' gestation results in both reduced bronchiolar branching and retarded acinar development, that, at late stage, influences only acinar development.

Amino Acids↗

Gastric antisecretory and antiulcer actions of interleukin-1. Evidence for the presence of an "immune-brain-gut" axis.

Increasing evidence suggests that interleukin-1 (IL-1), a cytokine mainly produced by activated monocytes/macrophages, has various biological actions in addition to its immunological activities. In the present study, we examined the effect of IL-1 on gastric secretion and gastric ulcer formation in rats. Gastric secretion was assessed in conscious pylorus-ligated rats weighing approximately 200 g. The peripheral injection of IL-1 resulted in a dose-related inhibition of gastric acid output. The central injection of IL-1 similarly reduced gastric acid secretion at 100 times smaller doses than peripherally injected IL-1, suggesting that this gastric antisecretory action of IL-1 is mediated by the central nervous system. In addition, it was found that this inhibitory effect of IL-1, either peripherally or centrally administered, was still evident at 8 h after injection, indicating the long-lasting property of this IL-1 action. On the basis of these antisecretory actions of IL-1, we determined whether or not pretreatment with IL-1 would prevent experimentally induced gastric ulcer formation. As expected, the central administration of IL-1 dose-dependently suppressed the development of gastric mucosal lesions induced by water-immersion restraint stress, a well-established ulcerogenic procedure. These results clearly demonstrated that IL-1 has potent antisecretory and antiulcer effects that are mediated by the central nervous system. Moreover, these findings suggest that there may exist an "immune-brain-gut" axis, which is involved in the regulation of gastric secretion and mucosal homeostasis, especially under certain pathophysiological conditions that activate the immune system to release various cytokines including IL-1.

Animals↗

Direct interfacing of high-speed countercurrent chromatography to frit electron ionization, chemical ionization, and fast atom bombardment mass spectrometry.

Direct interfacing of analytical high-speed countercurrent chromatography (HSCCC) to mass spectrometry (MS) was demonstrated for the first time, and its performance was evaluated in terms of chromatography and mass spectrometry. HSCCC/MS interface was based upon Frit electron ionization (EI), chemical ionization (CI), and fast atom bombardment (FAB). Separations were conducted by newly developed HSCCC-4000 with a 2.5-cm revolutional radius and 0.3 mm or 0.55 mm i.d. multilayer coiled column which is capable of operating at a maximum speed of 4000 rpm. To demonstrate the potential capability of HSCCC/frit MS, three indole auxin mixtures, two mycinamicin (macrolide antibiotics) mixtures, and a colistin complex (peptide antibiotics) were analyzed under HSCCC/frit EI, CI, and FABMS conditions, respectively. The data obtained indicated that interfacing to frit/MS does not adversely affect the chromatographic resolution and mass spectra provide structural information. The HSCCC system interfaced with a frit-equipped mass spectrometer will offer a new dimension in the separation of biologically important substances.

Chromatography↗

The effect of apo E secretion on lipoprotein uptake in transfected cells.

To investigate the role of apolipoprotein E (apo E) secreted by peripheral tissues in local lipoprotein metabolism, we developed a cell strain that constitutively produced and secreted apo E. A fusion plasmid containing rat apo E genomic DNA under control of mouse metallothionein promotor was constructed and transfected into Chinese hamster ovary cells. A stable transformant designated CHO-MAEII constitutively secreted rat apo E mainly in the form of sialylated free protein. The secretion was further enhanced by metal induction up to 1 micrograms apo E/ml per 12 h. When incubated with 125I-labeled very low density lipoprotein (125I-VLDL) at 37 degrees C, CHO-MAEII took up and degraded 125I-VLDL with higher affinity than control cells. Furthermore, considerable amount of methylated 125I-VLDL was degraded by CHO-MAEII, while no methylated 125I-VLDL was degraded by control cells. No significant differences were found in the uptake of 125I-LDL. The data indicated that apo E molecules secreted by CHO-MAEII were transferred to 125-VLDL particles, which caused a higher affinity of these particles for LDL receptors on the cells. It is suggested that apo E secreted from peripheral tissues enhances the uptake of lipoproteins by themselves or by surrounding cells in the local environment which demand cholesterol and express LDL receptors. CHO-MAEII was a good model for these 'auto- or paracrine-like functions' of apo E.

Animals↗

Cardioprotective effects of KRN2391 and nicorandil on ischemic dysfunction in perfused rat heart.

The cardioprotective effect of KRN2391 (N-cyano-N-(2-nitroxymethyl)-3- pyridinecarboximidamide methanesulfonate), a novel vasodilator, was studied in the isolated perfused rat heart and compared with that of nicorandil. The isolated buffer-perfused rat heart was subjected to 25 min ischemia followed by 30 min reperfusion. The heart was pretreated with 0.1-10 microM KRN2391, 10-1000 microM nicorandil or vehicle. Before ischemia, KRN2391 (1-10 microM) and nicorandil (10-1000 microM) increased coronary flow, but did not modify the cardiac mechanical function. KRN2391 (1-10 microM) and nicorandil at high doses (300-1000 microM) resulted in significant improvements of cardiac functions and coronary flow during reperfusion and significantly reduced the release of cytosolic enzymes. The protective effects of 3 microM KRN2391 and 300 microM nicorandil were completely reversed by 3 microM glibenclamide, a blocker of ATP-sensitive potassium channels. Thus, KRN2391 and nicorandil at high doses have a direct cardioprotective effect, which may be related to activation of ATP-sensitive potassium channels.

Animals↗

[Changes in the cartilaginous contour of Legg-Calvé-Perthes disease: calculation on T1-weighted MR images].

T1-weighted MR images of Legg-Calvé-Perthes disease (LCPD) were classified into three groups on the basis of radiographic stage, and morphological differentiation for staging was attempted. In the stage of fragmentation, both enlargement and flattening of the cartilaginous contour surrounding the epiphysis could be recognized on MRI, and the growth plate showed more curvature than normal. This produced flattening of the epiphysis in the shape of a crescent. We confirmed these findings using four indexes for the measurement of cartilaginous outline, and the stage of avascular necrosis and fragmentation could be clearly differentiated. Cartilaginous deformities on MRI are very useful for differentiating between the stage of avascular necrosis and fragmentation.

Cartilage, Articular↗

Excitation-contraction uncoupling during ischemia in the blood perfused dog heart.

The bioluminescent of Ca(2+)-indicator, aequorin, was loaded into the left ventricular apex of blood-perfused hearts from 13 dogs for simultaneous recording of left ventricular pressure and intracellular calcium levels. During a 2 minute period of ischemia, systolic and diastolic pressures significantly decreased. In contrast, these pressure changes were associated with an increase in both systolic and diastolic calcium reaching a maximum diastolic value of 0.59 microM and a systolic value of 1.11 microM. This apparent dissociation between pressure and [Ca2+]i supports the hypothesis that changes in myofilament Ca2+ responsiveness are of major importance in modulating contractility during ischemia in large mammalian hearts.

Aequorin↗

[MR imaging of osteoradionecrosis of the mandibula following radiotherapy for head and neck cancers].

Magnetic resonance (MR) images of 13 patients with osteoradionecrosis of the mandibula were analyzed. Abnormalities were noted in all patients and could be classified into three groups. In nine patients, MR signals were homogeneously low in intensity on both T1 weighted images (T1WI) and T2 weighted images (T2WI), suggesting fibrosis of bone marrow. These nine patients had suffered radiologically and/or clinically overt osteoradionecrosis more than ten months previously, and inflammatory symptoms had settled down by the time of MR study. Two patients showed low signal intensity on T1WI and an area of inhomogeneous high intensity in a diffuse area of low signal intensity on T2WI, suggesting acute inflammation in the irradiated fibrous bone marrow. Both patients suffered pathologic fracture due to osteoradionecrosis and showed severe inflammatory symptoms at the time MR study. In one of the two patients, these pathologic changes were confirmed in surgically excised mandibulectomy specimens. The other two patients showed homogeneous low signal intensity on T1WI and high signal intensity on T2WI, a finding that may be due to slight inflammation. However, in one of the two patients, this signal feature possibly indicated loose fibrosis with marked cellularity.

Adult↗

[MRI evaluation of the patellar articular cartilage in patients with subluxation of the patella].

In patients with subluxation of the patella, injury of the patellar articular cartilage is frequently observed and correct evaluation is important to manage these patients. We examined 11 patients with subluxation of the patella and five normal volunteers. In 12 patellofemoral joints of seven patients with subluxation of the patella, the abnormalities observed on MRI were compared with those on arthroscopy and/or at operation. MRI was performed with a Magnetom 1.5 T (Siemens) using the round surface coil. Pulse sequences were SE (TR 400 ms/TE 19 ms), FLASH (TR 320 ms/TE 15 ms FA 90 degrees and 40 degrees), and SE (TR 2000 ms/TE 26, 70 ms). We analysed MR findings of the 12 abnormal joints and 10 normal joints according to the following classification of abnormalities observed on arthroscopy. (1) normal appearance (n = 3 joints), (2) softening and fibrillation (n = 6), (3) fragmentation (n = 3), and (4) erosion to bone (n = 0). In only one of the six cases with softening and fibrillation observed on arthroscopy, MRI could visualize the thickening of patellar articular cartilage, but in all three cases with fragmentation observed on arthroscopy, MRI could visualize the thin inhomogenous cartilage with irregular surface. The combination of SE (TR 400 ms/TE 19 ms) and FLASH (TR 320 ms/TE 15 ms FA 90 degrees) are extremely effective pulse sequence to detect the abnormalities of patellar articular cartilage. We conclude that MRI is a useful noninvasive method of detecting advanced changes in patellar articular cartilage.

Adolescent↗

Improvement of chemical analysis of antibiotics. XVII. Application of an amino cartridge to the determination of residual sulphonamide antibacterials in meat, fish and egg.

A simple, rapid and reliable method for the determination of residual sulphonamide antibacterials (SAs) (sulphathiazole, sulphisozole, sulphamethoxazole, sulphadiazine, sulphamerazine, sulphadimidine, sulphamonomethoxine, sulphadimethoxine, sulphamethoxypyridazine and sulphaquinoxaline) in meat, fish and egg was developed using a combination of high-performance liquid chromatography (HPLC) and clean-up with an amino-type prepacked cartridge. SAs were extracted with ethyl acetate and applied to a Baker 10 amino cartridge. After elution from the cartridge, SAs were determined by HPLC. The recoveries at the level of 0.5 ppm were 73.7-99.1% and the detection limits were 0.05 ppm. The analysis time per sample was about 45 min.

Anti-Bacterial Agents↗

[Dynamic MRI of hepatocellular carcinoma with portal vein tumor thrombus].

Dynamic MRI with injection of Gd-DTPA was performed in 7 patients with hepatocellular carcinoma associated with portal vein tumor thrombus. A portal vein tumor thrombus was clearly visualized as a low intense structure. The segment supplied with this portal vein shows high intensity, probably due to the decrease in portal blood flow and compensated arterial blood flow. A tumor itself, located in this segment, was shown as a low intense area. Thus dynamic MRI is a useful method for differentiating the tumor from the surrounding nontumorous tissue with decrease in portal flow.

Aged↗

Enhanced replication of herpes simplex virus by hexamethylene bisacetamide.

Hexamethylene bisacetamide (HMBA) is a potent inducer of differentiation of tumor cells. The effect of HMBA on cell growth and replication of herpes simplex virus (HSV) was investigated in HEp-2 epidermal cells, IMR-32 neuronal cells, K562 myeloid cells, Daudi Burkitt lymphoma cells, and CCRF-CEM T-lymphoid cells. The growth of HEp-2 and IMR-32 cells was not affected by HMBA at concentrations from 0.5 through 2 mM. The growth of K562, Daudi, and CCRF-CEM cells was inhibited by HMBA at concentrations from 1 through 5 mM. When HSV-infected cells were incubated with 0.5 through 5 mM HMBA, a dose-dependent increase in virus yield was observed in HEp-2 and IMR-32 cells, but not in the other cell lines. These findings indicate that HMBA enhances the replication of HSV in epidermal and neuronal cells and that HMBA therapy may be responsible for the development of herpetic lesions.

Acetamides↗

Lack of feedback inhibition of V kappa gene rearrangement by productively rearranged alleles.

Circular DNAs excised by immunoglobulin kappa chain gene rearrangements were cloned and characterized. 16 of 17 clones examined were double recombination products containing a V kappa-J kappa rearrangement (coding joint) as well as the reciprocal element (signal joint) of another V kappa-J kappa rearrangement. These products suggested multiple recombination, primary inversion, and secondary excision. In primary events, 5 of 16 translational reading frames were in-phase. Thus, V kappa gene rearrangement may not be inhibited by the presence of a productively rearranged allele. An unusually large trinucleotide (P) insertion forming a palindrome of 12 nucleotides was also observed in one of the coding joints.

Alleles↗

Purification of Food Color Red No. 106 (acid red) using high-speed counter-current chromatography.

High-speed counter-current chromatography (HSCCC) has been successfully applied to the separation of the components of Food Color Red No. 106 (R-106). The separation was performed using 25 mg of the sample with a two-phase solvent system composed of n-butanol and 0.01 M trifluoroacetic acid (1:1, v/v). Analyses by thin-layer chromatography, high-performance liquid chromatography and fast atom bombardment mass spectrometry confirmed that HSCCC was effective in the purification of the components of R-106. The separation gave 21 mg of a 99.9% pure main component (Acid Red) and 0.9 mg of 98.0% pure subsidiary dye which is probably a des-ethyl derivative.

Azo Compounds↗

Isolation of bacitracins A and F by high-speed counter-current chromatography.

The major components of bacitracin were separated and purified using high-speed counter-current chromatography (HSCCC). A systematic search for optimum two-phase solvent systems resulted in two systems: chloroform-ethanol-methanol-water (5:3:3:4) and chloroform-ethanol-water (5:4:3). These were selected based on the determination of the partition coefficients of all the components and the settling time of the phases. HSCCC with these solvent systems separated two components, bacitracins A and F. Improvements in the flow-cell arrangement eliminated noise in detection, making in-line monitoring possible. A tandem mass spectrometric technique was used to characterize the isolated components.

Bacitracin↗