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Biomedical subjects

K Horiuchi

Publications and source records attributed to K Horiuchi.

At least 37 records · Page 2Linked to original sources

Direct detection of crosslinks of collagen and elastin in the hydrolysates of human yellow ligament using single-column high performance liquid chromatography.

Collagen and elastin are recognized as two major connective tissue proteins of human yellow ligament. In both collagen and elastin there are many kinds of intra- or intermolecular crosslinks. Pyridinoline (Pyr) and deoxypyridinoline (Dpyr) are mature crosslinks which maintain the structure of the collagen fibril. Desmosine (Des) and isodesmosine (Isodes) represent the major crosslinking components of elastin. Pentosidine (Pen), which is a senescent crosslink and one of the advanced glycation end products, accumulates with age in tissue proteins including collagen. We developed a direct and one-injection HPLC method to measure Pyr, Dpyr, Des, Isodes, and Pen in the hydrolysate of human yellow ligament. This method used one column and two detectors. Recovery rates of Pyr, Dpyr, Pen, Des, and Isodes were 86.4-98.3, 83.6-96.8, 78.7-95.6, 83.6-97.9, and 85.6-99.3%, respectively (n = 8). The intraassay coefficients of variation for Pyr, Dpyr, Pen, Des, and Isodes were 3.7, 4.1, 5.4, 4.5, and 4.7%, respectively (n = 8), and the interassay coefficients of variation for Pyr, Dpyr, Pen, Des, and Isodes were 4.4, 5.1, 4.9, 4.6 and 4.1%, respectively. Linear regression analysis showed the linearity (r = 0.99, P = 0.0001) of calibration line for each Pyr, Dpyr, Pen, Des, and Isodes. Using this method, we investigated age-related changes in the crosslinks of collagen and elastin in human yellow ligament. There was a significant correlation between Pen and age, but no correlations with Pyr, Dpyr, Des, and Isodes. We believe that this method is useful for investigating the content of these crosslinks in both collagen and elastin under various conditions.

Chromatography, High Pressure Liquid↗

Enhancement of hemoglobin and F-cell production by targeting growth inhibition and differentiation of K562 cells with ribonucleotide reductase inhibitors (didox and trimidox) in combination with streptozotocin.

Upon appropriate drug treatment, the human erythroleukemic K562 cells have been shown to produce hemoglobin and F-cells. Fetal hemoglobin (Hb F) inhibits the polymerization events of sickle hemoglobin (Hb S), thereby ameliorating the clinical symptoms of sickle cell disease. Ribonucleotide reductase inhibitors (RRIs) have been shown to inhibit the growth of myeloid leukemia cells leading to the production of Hb F upon differentiation. Of the RRIs currently in use, hydroxyurea is the most effective agent for Hb F induction. We have examined the capacity of two novel RRIs, didox (DI) and trimidox (TRI), in combination with streptozotocin (STZ), to induce hemoglobin and F-cell production. The K562 cells were cultured with different concentrations of didox-STZ or trimidox-STZ at a fixed molar ratio of 3:1 and 1:5 for 96 hr, respectively. At pre-determined time intervals, aliquots of cells were obtained and total hemoglobin (benzidine positive) levels, number of F-cells, and Hb F were determined by the differential staining technique, fetal hemoglobin assay kit, and fluorescence cytometry respectively. The effect of combined drug treatment on the growth of K562 cells was examined by isobologram analysis. Our results indicate that a synergistic growth-inhibitory differentiation effect occurred when didox or trimidox was used in combination with STZ on K562 cells. There was an increase in the number of both benzidine-positive normoblasts and F-cells, accompanied by morphologic appearances typical of erythroid maturation. On day 4, the number of benzidine-positive cells showed a 6-9-fold increase and the number of F-cells was between 2.5- and 5.7-fold higher than the respective controls. Based upon these results, treatment with a ribonucleotide reductase inhibitor, such as didox or trimidox, in combination with STZ, might offer an additional promising option in sickle cell disease therapy.

Antineoplastic Agents↗

The development of (99m)Tc-analog of Cu-DTS as an agent for imaging hypoxia.

Works on dithiosemicarbazone (DTS) derivatives radiolabeled with divalent Cu (Cu-62, Cu-64) indicate its potentiality as an ischemic tissue detecting agent. Development of analogous derivatives labeled with the more accessible technetium-99m (Tc) is most desirable. Various synthesized DTS derivatives are radiolabeled with a novel approach, using a macromolecular Sn(II)-complex under an anaerobic condition at pH 3.4-4.5 and stabilization by ascorbate solution at pH 6.7-7.0. Characterization of Tc-DTS derivatives done by various analytical methods (TLC, HPLC, EP, PC) and by in vivo studies in normal mice and in rats myocardial LAD (left anterior descent coronary artery) occlusion model. Among tested DTS, only Tc-ATSE, Tc-ATSM and Tc-ATSM(2) showed distinctive characteristics, with the latter presenting high myocardium uptake in regions of ischemia in LAD rat myocardium model. Potentiality of the Cu-DTS mimetic agent, Tc-ATSM(2) as an ischemia-damaged myocardium agent is discussed.

Animals↗

Renal masses detected by general health checkup.

BACKGROUND: A total of 60,604 persons underwent a general health checkup at Toma Hospital, Saitama, Japan, between January 1993 and June 1997, and transabdominal ultrasonography (US) was performed on all persons. We investigated the usefulness of transabdominal US in detecting renal tumors during general health checkups. METHODS: A definite diagnosis was made in cases where the re-examination by US revealed calculus with acoustic shadow or a simple cyst. Computed tomography (CT) was conducted in patients suspected of having a renal tumor, and a diagnosis of renal angiomyolipoma (AML) was made if plain CT gave evidence of a fatty component. Patients diagnosed as having a renal cell carcinoma (RCC) with non-uniform enhancement underwent nephrectomy. If a definite diagnosis could not be made even by contrast CT, the monitoring by US and contrast CT once a year was continued. RESULTS: Of 97 (0.16%) patients whose transabdominal US findings indicated a diagnosis of suspected renal tumor, 58 underwent the re-examination by US; of these, CT was conducted in 47. Detailed US revealed a hyperechoic pattern in 44 patients, a mixed pattern in nine and an isohypoechoic pattern with internal echo in five. By plain CT, 24 patients were diagnosed as having AML, and two as having renal calcification. Contrast CT enabled a diagnosis of renal cyst to be made in two patients and renal tumor in 14. All 14 patients with RCC except one underwent nephrectomy, which provided pathologic evidence supporting the diagnosis. Five patients had a tumor unidentifiable by CT, with continuing periodic follow-up by US and CT. Eleven patients underwent no CT. CONCLUSION: Of the 60,604 persons who underwent a general health checkup, we found 14 cases of RCC and 24 cases of AML. As a small RCC has a favorable prognosis, patients greatly benefit from early detection; therefore, we should recognize the importance of identifying renal masses by US screening during general health checkups.

Adolescent↗

Correlation between deposition of immuno-components and infiltration pattern of polymorphonuclear leukocytes in the lesions of chronic urticaria.

Urticaria is often associated with perivascular infiltration of leukocytes into the lesions. Although mast cell-derived chemical mediators are considered to play crucial roles in the infiltration of leukocytes as well as in the dermal edema, other mechanisms for the leukocyte infiltration have not been well defined. This study revealed that approximately 25% of the cases of chronic idiopathic urticaria in whom wheals had continued for more than 12 h had deposition of immuno-components in the lesions, although histological examination of the lesions did not show leukocytoclastic vasculitis. In these lesions with deposition of immuno-components, both neutrophils and eosinophils had infiltrated at a constant ratio (approximately 2:1), whereas, in the lesions without deposition, a variable population of leukocytes was seen. This result suggests that activation of complements occurs in the lesions of a considerable percentage of patients with chronic idiopathic urticaria and that the complement fragments influence the infiltration patterns of polymorphonuclear leukocytes.

Adult↗

Endoscopic treatment of vesicoureteral reflux in children with glutaraldehyde cross-linked bovine dermal collagen. Short-term results.

BACKGROUND: Endoscopic treatment using glutaraldehyde cross-linked (GAX) collagen was conducted on 4 children with bilateral primary vesicoureteral reflux (VUR)..... a 1-year-old boy and three 5- to 8-year-old girls $B!D (Ball having a history of repeated hospitalization for fever due to acute pyelonephritis, visiting as pediatric outpatients regularly and receiving antibiotics continuously. By international VUR classification, 4 ureters were grade 3, 2 grade 4, and 2 grade 5. METHODS: After nonallergy to GAX collagen was confirmed intracutaneously, a needle was used through a 9.5 Fr cystoscope channel to puncture bladder mucosa 4 to 5 mm from the affected ureteral orifice at 6 o'clock under general anesthesia; 1.1 to 1.9 ml of GAX collagen was injected immediately below affected orifices. RESULTS: Three months after surgery, voiding cystourethrography showed reflux had disappeared in 6 ureters, for a short-term success rate of 75%. VUR in the Remaining 2 ureters improved from grade 3 to 1 and from grade 5 to 4. No postoperative urinary tract infection occurred and antibiotics were stopped. CONCLUSION: Since GAX collagen is less viscous than Teflon paste, it is easily injected into submucosa, does not form granuloma or migrate to other organs, and is noncarcinogenic. Endoscopic VUR treatment using GAX collagen is indicated when less invasion and shorter hospitalization are considered, although it requires general anesthesia, which itself involves some risk.

Child↗

[A case of tuberculous epididymitis which was difficult to distinguish from a testicular tumor].

A 70-year-old man complaining of painless right scrotal swelling was referred to our hospital. He had a past history of left nephrectomy for renal tuberculosis at the age of 28. Power Doppler ultrasonography revealed a hypoechoic and hypovascular tumor with septa in the scrotum. We suspected a right testicular tumor and therefore, performed a right high inguinal orchiectomy. On macroscopic findings, the fluid of the tumor was yellowish and mucinous and the ipsilateral testis was remarkably atrophic. The pathological diagnosis was tuberculous epididymitis with central necrosis. In recent years, tuberculous epididymitis is rare, and this case was considered to be the first report of power Doppler ultrasonographic findings in tuberculous epididymitis.

Aged↗

Evaluation of threshold stress for bone resorption around screws based on in vivo strain measurement of miniplate.

The purpose of this study is to investigate the critical threshold stress causing bone resorption evaluated from strain measurement in vivo, comparing the various finite element models. In this study strains of miniplates used for mandibular fractures were measured once a week until the strains reduced. The maximum bite force for each patient was applied in the incisal, right molar and left molar region. The strains increased and reached a peak level at 2-4 weeks, whereas the bite forces increased during the period of measurements. A 3-D osteosynthesis model using finite element method showed that the compressive stresses of the bone surrounding screws ranged within approximately -40 MPa under the condition generating the same amounts of strains measured in the miniplates. Furthermore, various finite element models simulating mandibular reconstruction using the fibular graft were constructed. The models for reconstruction using single strut fibula showed distinct stress concentration in the cortical bone surrounding screws, and the peak stress levels were 2 to 3 times as strong as that of the fracture model. We conclude that critical threshold for bone resorption should be approximately -50 MPa (3600 micro strain).

Journal Article↗

Patch clamp studies on V-type ATPase of vacuolar membrane of haploid Saccharomyces cerevisiae. Preparation and utilization of a giant cell containing a giant vacuole.

A method for obtaining giant protoplasts of Escherichia coli (the spheroplast incubation (SI) method: Kuroda et al. (Kuroda, T., Okuda, N., Saitoh, N., Hiyama, T., Terasaki, Y., Anazawa, H., Hirata, A., Mogi, T., Kusaka, I., Tsuchiya, T., and Yabe, I. (1998) J. Biol. Chem. 273, 16897-16904) was adapted to haploid cells of Saccharomyces cerevisiae. The yeast cell grew to become as large as 20 micrometer in diameter and to contain an oversized vacuole inside. A patch clamp technique in the whole cell/vacuole recording mode was applied for the vacuole isolated by osmotic shock. At zero membrane potential, ATP induced a strong current (as high as 100 pA; specific activity, 0.1 pA/micrometer(2)) toward the inside of the vacuole. Bafilomycin A(1,) a specific inhibitor of the V-type ATPase, strongly inhibited the activity (K(i) = 10 nM). Complete inhibition at higher concentrations indicated that any other ATP-driven transport systems were not expressed under the present incubation conditions. This current was not observed in the vacuoles prepared from a mutant that disrupted a catalytic subunit of the V-type ATPase (RH105(Deltavma1::TRP)). The K(m) value for the ATP dose response of the current was 159 microM and the H(+)/ATP ratio estimated from the reversible potential of the V-I curve was 3.5 +/- 0.3. These values agreed well with those previously estimated by measuring the V-type ATPase activity biochemically. This method can potentially be applied to any type of ion channel, ion pump, and ion transporter in S. cerevisiae, and can also be used to investigate gene functions in various organisms by using yeast cells as hosts for homologous and heterogeneous expression systems.

Anti-Bacterial Agents↗

Filamentous phage replication initiator protein gpII forms a covalent complex with the 5' end of the nick it introduced.

Rolling circle type DNA replication is initiated by introduction of a nick in the leading strand of the origin by the initiator protein, which in most cases binds covalently to the 5' end of the nick. In filamentous phage, however, such a covalent complex has not been detected. Using a suitable substrate and short reaction time, we show that filamentous phage initiator gpII forms a covalent complex with nicked DNA, which rapidly dissociates unless gpII is inactivated. A peptide-DNA complex was isolated from trypsin digest of the complex by ion-exchange column chromatography and gel filtration, and its peptide sequence was determined. The result indicated that gpII was linked to DNA by the tyrosine residue at position 197 from the N-terminus. The mutant protein in which this tyrosine was replaced by phenylalanine did not show any detectable activity to complement gene II amber mutant phage in vivo. In vitro, the mutant protein recognized the origin and bent DNA as well as the wild-type does, but failed to introduce a nick and to relax the superhelicity of cognate DNA.

Coliphages↗

Clinical utility of negative contrast intravascular ultrasound to evaluate plaque morphology before and after coronary interventions.

Although intravascular ultrasound (IVUS) is used for evaluation of plaque volume and lumen size as well as detection of vessel wall structures after catheter-based interventions, differentiation between the lumen and plaque structures can be difficult. This study attempted to evaluate the efficacy of negative contrast IVUS imaging for assessment of vessel wall morphology after coronary interventions. IVUS studies were performed in 67 lesions in 66 patients before and after coronary interventions. After the baseline ultrasound imaging run, warm 5% glucose solution was injected manually through the guiding catheter into the coronary artery to washout blood from the lumen to avoid speckled reflections from red blood cells (negative contrast). Quantitative measurements were obtained and plaque morphology was assessed for the presence and extent of medial dissections and intimal flaps. There was no difference in each quantitative parameter between baseline images and negative contrast images. The vessel wall boundary was clearly delineated from the lumen, which was defined as effective negative contrast in 51 of 67 lesions (76%). The baseline images revealed plaque dissection in 9 lesions (18%) and an intimal flap in 13 lesions (25%). In addition, 4 dissections (8%) and 16 intimal flaps (31%) were visualized during the infusion of negative contrast. Additional treatment was performed in 4 lesions (8%) based on the images with negative contrast. Negative contrast IVUS was more sensitive in demonstrating a plaque fracture than were baseline images. This method is useful for enhancing the diagnostic capability of IVUS imaging and may influence the decision-making process during interventional procedures.

Anatomy, Cross-Sectional↗

Emerging and entraining patterns of the sleep-wake rhythm in preterm and term infants.

It has been repeatedly reported that the sleep-wake rhythm in infants entrains around 3-4 months of age after a transient free-run rhythm. To clarify the emerging and entraining patterns of the sleep-wake rhythm, the sleep and wakefulness of 84 infants (44 preterm and 40 term infants) were longitudinally recorded at home for more than 16 weeks by the day-by-day plot method. Our results showed that the entrained sleep-wake rhythm emerged after transient manifestation of either ultradian or irregular sleep-wake patterns for 3-4 weeks in 75% of the infants. Only 7% of the infants showed a free-running sleep-wake rhythm before the entrainment. These facts suggest that most infants would be entrained to an ordinary daily schedule of mothers without expression of overt free-running rhythm of the biological clock. The mean age of the entrainment was 44.8 postconceptional weeks. There were no significant differences in either frequency of each pattern or the mean age of the entrainment, between preterm and term infants. In conclusion, the entrained sleep-wake rhythm emerges around 1 corrected month, after ultradian patterns in the majority of infants.

Circadian Rhythm↗

Carrier effect on radiolabeling the polynuclear pentavalent rhenium-186 complex of dimercaptosuccinic acid at alkaline pH: 186Re(V)-DMS.

Radiolabeling with rhenium (Re-186, Re-188), a tumor agent to resemble the pentavalent polynuclear technetium complex of dimercaptosuccinic acid (99mTc[V]-DMS) has been reported for radiotherapeutical use. Nevertheless, despite the periodic analogies between both radiometals, differences in the redox potential and the carrier concentration have made the radiolabeling of the rhenium counterpart difficult. In the present study, the effect of the carrier contained in the reactor-produced Re-186 was estimated as an important factor relevant to the Re-186 radiolabeling of DMS at an alkaline pH. Great effect of the carrier Re with an inverse correlation with the stannous ion was an interesting phenomenon relevant for an assumption on the Sn participation in the complex. Under strict control of various labeling parameters, the 186Re(V)-DMS was made available with high yield (93-97%) at an alkaline pH and at room temperature. The great effect of carrier offers support to the polymeric or polynuclear nature of the rhenium complex of DMS as depicted in the drug design basis of its parent Tc(V)-DMS. The biodistribution studies of Re(V)-DMS showed mimetic characteristics with its parent Tc(V)-DMS drug.

Animals↗

The integrity of the disulfide bond in a cyclic somatostatin analog during 99mTc complexation reactions.

Recent development of a variety of thiol-free chelating agents has facilitated the design of 99mTc-labeled somatostatin analogs suitable for receptor imaging of somatostatin-positive tumors. However, it remains ambiguous whether the disulfide bonds in cyclic peptides are stable during 99mTc complexation reactions, and contradictory results have been reported regarding the integrity of disulfide bonds in cyclic somatostatin analogs. To estimate the stability of the disulfide bond in a synthetic somatostatin analog at low peptide concentrations, [125I]I-RC-160, in which radioiodine was incorporated into the 3-Tyr residue, was synthesized and the integrity of the disulfide bond of the peptide was investigated in the presence of reducing agents such as ascorbic acid, dithionite, and stannous ions. The disulfide bond in [125I]I-RC-160 remained stable in the presence of ascorbic acid in boiling water. The disulfide bond was also stable when treated with stannous ions at concentrations sufficient to reduce 99mTc for complexation with a thiol-free chelating agent, bis(hydroxamamide) analog when the 99mTc complexation reaction was performed at room temperature. However, the disulfide bond of [125I]I-RC-160 was slightly cleaved in the presence of a small amount of stannous ions when the reaction was performed in boiling water. Treatment of [125I]I-RC-160 with dithionite in boiling water markedly reduced the disulfide bond of the parental peptide. These findings indicated that synthetic somatostatin analogs may be labeled with 99mTc with stannous ions as the reducing agent without impairing their structure after conjugation of thiol-free chelating agents that provide 99mTc chelates under mild reaction conditions.

Ascorbic Acid↗

Influence of lower limb pneumatic compression on pulmonary artery temperature: effect on cardiac output measurements.

OBJECTIVES: To characterize the decreases in pulmonary artery temperature that coincide with the inflation cycle of pneumatic calf compression stockings and to examine their effects on the thermodilution measurement of cardiac output. DESIGN: Three-part observational study. SETTING: University hospital surgical intensive care unit. PATIENTS: Postoperative patients with indwelling pulmonary artery catheters. INTERVENTION: Thermodilution cardiac output measurements with and without pneumatic calf compression. MEASUREMENTS AND MAIN RESULTS: Phase 1 (n = 18) examined the effects of pneumatic compression on pulmonary artery temperature. There was no effect on pulmonary artery temperature (device off, 37.468+/-0.008 degrees C; device on, 37.458+/-0.014 degrees C), but the difference between the maximum and minimum pulmonary artery temperatures was increased (off, 0.031+/-0.006 degrees C; on, 0.055+/-0.012 degrees C [p < .001]). Phase 2 (n = 12) found that the mean thermodilution cardiac output with 10 mL of cold (0-5 degrees C) injectate was unchanged by pneumatic compression (off, 7.00+/-2.28 L/min; on, 6.89+/-2.22 L/min). However, when the compression devices were operating, the variability between the individual measurements was increased, as reflected by larger coefficients of variation (off, 3.19+/-1.96; on, 8.72+/-6.56 [p < .02]). Similar results were obtained during phase 3 (n = 5), when cardiac output was measured with room temperature Injectate. CONCLUSIONS: Intermittent pneumatic calf compression increased lower limb venous return, causing acute but transient decreases in pulmonary artery blood temperature. This did not affect the accuracy of thermodilution cardiac output measurements that were made using 10 mL of either cold or room temperature injectate.

Bandages↗

A case of acquired autoimmune bullous disease associated with IgM macroglobulinaemia.

A variety of autoimmune bullous dermatoses have been reported to develop in association with lymphoproliferative disorders. We report a patient with IgM macroglobulinaemia, who presented with a skin fragility similar to but somewhat milder than that seen in epidermolysis bullosa acquisita. Immunofluorescence detected circulating IgM autoantibodies reacting with the basement membrane zone, which reacted predominantly with dermal side of 1 M NaCl-split skin. Immunoblotting of the epidermal and dermal extracts with the patient's serum showed no specific reactivity. Further studies are needed to identify the antigenic molecules responsible for the IgM deposition.

Autoantibodies↗

Elimination of non-ionic contrast medium by hemodialysis in patients with impaired renal function.

The elimination rate of iohexol, a non-ionic contrast medium, from the blood by hemodialysis, and the elimination rate of iohexol by a dialyzer were studied in 15 patients with chronic renal dysfunction who required angiography or enhanced CT. The elimination rate of iohexol was 19.8% at 15 min after the start of hemodialysis, 30.6% after 30 min, 44.2% after 1 hour, 62.1% after 2 hours and 72.9% after 3 hours. The dialyzer elimination rate was maintained at about 75% from 1 to 3 hours after the start of hemodialysis. If only about 70% of iohexol in the blood needs to be eliminated, hemodialysis for 3 hours with a blood flow rate of 120 ml/min and a dialysate flow of 500 ml/min using a 0.7 m2 cellulose triacetate membrane is sufficient.

Contrast Media↗