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Biomedical subjects

K Takayama

Publications and source records attributed to K Takayama.

At least 109 records · Page 6Linked to original sources
PubMed1999

Distribution characteristics of entrapped recombinant human erythropoietin in liposomes and its intestinal absorption in rats.

Recombinant human erythropoietin (Epo) is frequently administered by intravenous (i.v.) injection for the clinical treatment of renal anemia. Oral (per os; p.o.) administration is desired as an alternative route to i.v. administration, and liposomes have been chosen as a drug carrier. We found previously that after a p.o. administration to rats of Epo entrapped in liposomes before gel filtration, the Epo was absorbed, but variability in the number of days of appearance and in the levels of pharmacological effects, i.e. , the peak of circulating reticulocyte counts (RTC), was observed. The purpose of the present study was to examine the distribution characteristics of Epo in liposomes and intestinal absorption of liposomal Epo in rats by using purified Epo entrapped in liposomes after gel filtration (Epo/liposomes). The distribution characteristics of Epo/liposomes were determined by measuring the Epo in liposomes by a radioimmunoassay, high-performance liquid chromatography and zeta potential measurements. We observed that the protein part of Epo was mostly entrapped in liposomes, and was not adsorbed by the liposomal membrane at middle and high Epo p.o. doses, but the zeta potential of the Epo/liposomes increased negatively with the increase in the Epo p.o. doses. These results suggest that the sialic acid part of Epo entrapped in liposomes may project out from liposomes, depending on the entrapped Epo concentration. Little Epo was adsorbed or penetrated into liposomes when it was added to empty liposomes. After the p. o. administration of Epo/liposomes, the peak of RTC appeared at a 2-day delay on day 6, without variation and without dose dependency in comparison with that after i.v. administration. These results suggest that one of the reasons for the variability may be because the non-entrapped Epo and/or Epo/liposomes itself affected the intestinal absorption of Epo/liposomes. In conclusion, Epo/liposomes without nonentrapped Epo may be clinically useful for the oral administration of Epo.

Administration, Oral↗
PubMed1999

Absorption of insulin from pluronic F-127 gels following subcutaneous administration in rats.

The main objective of this work was to evaluate the use of Pluronic (PF127) gels, polylactic-co-glycolic acid (PLGA) nanoparticles and their combination for parenteral delivery of peptides and proteins having short half-lives using insulin as a model drug. The in vitro insulin release profiles of various PF127 formulations were evaluated at 37 degrees C using a membraneless in vitro model. In vivo evaluation of the serum glucose and insulin levels was performed following subcutaneous administration of various insulin formulations in normal rats. The in vitro results demonstrated that the higher the concentration of PF127 in the gel, the slower the release of insulin from the matrices, independent of the vehicle used. The acute hypoglycemic peak resulting from administration of an insulin solution between 0.5 and 2.0 h after administration (peak at 1 h) is replaced after administration of insulin-PLGA nanoparticles by an almost constant hypoglycemic effect with a slower recovery of the serum glucose levels at about 2 h after administration. By loading insulin into PF127 gels, a slower and more prolonged hypoglycemic effect of insulin was obtained in inverse proportion to the polymer concentration. PF127 gel formulations containing insulin-PLGA nanoparticles had the most long-lasting hypoglycemic effects of all formulations. From the current in vitro and in vivo study, we concluded that PF127 gel formulations containing either drug or drug-nanoparticles could be useful for the preparation of a controlled delivery system for peptides and proteins having short half-lives.

Animals↗
PubMed1999

Enhanced rectal absorption of insulin-loaded Pluronic F-127 gels containing unsaturated fatty acids.

The objective of this study was to prepare and to evaluate Pluronic F-127 (PF127) gel containing unsaturated fatty acids such as oleic acid (18:1), eicosapentaenoic acid (20:5) and docosahexaenoic acid (22:6) as a potential formulation for rectal delivery of insulin. The hypoglycemic effect of insulin was examined following rectal administration of the various formulations in normal rats. Rectal insulin absorption was markedly enhanced, and marked hypoglycemia was induced by all PF127 gels (insulin dose, 5 U/kg) containing different unsaturated fatty acids. PF127 gels containing unsaturated fatty acids presented low tmax mean values indicating that the absorption of insulin occurred very rapidly in the rectum. The relative hypoglycemic efficacy of PF127 gel formulations containing fatty acids such as oleic acid, eicosapentaenoic (EPA) and docosahexaenoic (DHA) were 28.4+/-8.1, 26.8+/-14.3 and 23.1+/-5.7%, respectively. The finding demonstrated that 20% PF127 gels containing unsaturated fatty acids are potential formulations for rectal delivery of insulin.

Administration, Rectal↗
PubMed1999

Remote loading of diclofenac, insulin and fluorescein isothiocyanate labeled insulin into liposomes by pH and acetate gradient methods.

Remote loading of the model drugs diclofenac, insulin and fluorescein isothiocyanate labeled insulin (FITC-insulin) into liposomes by formation of transmembrane gradients were examined. A trapping efficiency of almost 100% was obtained for liposomal diclofenac, by the calcium acetate gradient method, whereas liposomes prepared by the conventional reverse-phase evaporation vesicle method had 1-8% trapping efficiencies. Soybean-derived sterol was a better stabilizer of the dipalmitoylphosphatidylcholine bilayer membrane than cholesterol, as shown from trapping efficiencies and drug release. The pH gradient method resulted in a 5-50% of FITC-insulin liposomal trapping efficiency, while insulin could not be loaded by this method. Liposomes released calcein in response to insulin, showing insulin interacts with the liposomal membrane in the presence of a transmembrane gradient. The present work has demonstrated a remote loading method for weak acids such as diclofenac into liposomes by the acetate gradient method. From the result of remote loading of FITC-insulin into liposomes by the pH gradient method, this method may be available for the preparation of liposomal peptides.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed1999

Permeation of several drugs through keratinized epithelial-free membrane of hamster cheek pouch.

The hamster cheek pouch mucosa was selected as a substitute for the human buccal mucosa in an in vitro permeation study. Considering that a keratinized layer is not present in the human buccal mucosa, keratinized epithelial-free hamster cheek pouch (KEF-membrane) was prepared by chemical splitting. To confirm the usefulness of the KEF-membrane, a permeation study was conducted using several drugs with different lipophilicities. The permeability coefficient of hydrophilic drugs through the KEF-membrane (Pkef) was significantly greater than that through a viable KEF-membrane (Pkef-viable), which was estimated by using the permeability coefficient of the viable full-thickness membrane and that of the keratinized layer. On the other hand, the Pkef values of lipophilic drugs were comparable with the Pkef-viable values. Furthermore, the ratio of these P values (Pkef/Pkef-viable) decreased with increasing lipophilicity of drugs. These findings indicate that the KEF-membrane may be useful for buccal permeation studies of lipophilic drugs.

Animals↗
PubMed1999

Tetanus after a resection for a gangrenous perforated small intestine: report of a case.

We report herein the case of a 75-year-old man who developed severe tetanus 24 h after the resection of a gangrenous perforated small intestine. It seemed that the tetanus was caused by a spillage of the intestinal contents harboring Clostridium tetani; however, this was not identified by a culture. The diagnosis of tetanus was made only when opisthotonus in this patient became evident and normal tetanus treatment proved to be successful.

Aged↗
PubMed1999

Liquid jets, accelerated thrombolysis: a study for revascularization of cerebral embolism.

A prior study has reported that a rapid recanalization therapy of cerebral embolism, using liquid jet impacts generated by the interaction of gas bubbles with shock waves, can potentially penetrate through thrombi in as little as a few microseconds with very efficient ablation (Kodama et al. 1997). The present study was undertaken to examine the liquid jet impact effect on fibrinolysis in a tube model of an internal carotid artery. First, the conditions for generating the maximum penetration depth of liquid jets in the tube were investigated. Gelatin was used to mimic thrombi. The shock wave was generated by detonating a silver azide pellet weighing about a few micrograms located in a balloon catheter. The collapse of the inserted gas bubbles and the subsequent liquid jet formation were recorded with high-speed photography. Second, thrombi were formed using fresh human blood from healthy volunteers. The fibrinolysis induced by the liquid jet impact with urokinase was explored. This was conducted under selected conditions based on the experiment using the gelatin. Fibrinolysis was calculated as the percentage of the weight loss of the thrombus. Fibrinolysis with urokinase alone and with a single liquid jet impact with urokinase was 1.9 +/- 3.7% (n = 16) and 20.0 +/- 9.0% (n = 35), respectively, for an incubation time of 60 min. Statistical differences were obtained between all groups (ANOVA). These results suggest that liquid jet impact thrombolysis has the potential to be a rapid and effective therapeutic modality in recanalization therapy for patients with cerebral embolism and other clinical conditions of intra-arterial thrombosis.

Carotid Artery, Internal↗
PubMed1999

Artificial neural network as a novel method to optimize pharmaceutical formulations.

One of the difficulties in the quantitative approach to designing pharmaceutical formulations is the difficulty in understanding the relationship between causal factors and individual pharmaceutical responses. Another difficulty is desirable formulation for one property is not always desirable for the other characteristics. This is called a multi-objective simultaneous optimization problem. A response surface method (RSM) has proven to be a useful approach for selecting pharmaceutical formulations. However, prediction of pharmaceutical responses based on the second-order polynomial equation commonly used in RSM, is often limited to low levels, resulting in poor estimations of optimal formulations. The aim of this review is to describe the basic concept of the multi-objective simultaneous optimization technique in which an artificial neural network (ANN) is incorporated. ANNs are being increasingly used in pharmaceutical research to predict the non-linear relationship between causal factors and response variables. The usefulness and reliability of this ANN approach is demonstrated by the optimization for ketoprofen hydrogel ointment as a typical numerical example, in comparison with the results obtained with a classical RSM approach.

Chemistry, Pharmaceutical↗
PubMed1999

Granulocyte, granulocyte-macrophage, and macrophage colony-stimulating factors can stimulate the invasive capacity of human lung cancer cells.

We and other researchers have previously found that colony-stimulating factors (CSFs), which generally include granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF) and macrophage colony-stimulating factor (M-CSF), promote invasion by lung cancer cells. In the present study, we studied the effects of these CSFs on gelatinase production, urokinase plasminogen activator (uPA) production and their activity in human lung cancer cells. Gelatin zymographs of conditioned media derived from human lung adenocarcinoma cell lines revealed two major bands of gelatinase activity at 68 and 92 kDa, which were characterized as matrix metalloproteinase (MMP)-2 and MMP-9 respectively. Treatment with CSFs increased the 68- and 92-kDa activity and converted some of a 92-kDa proenzyme to an 82-kDa enzyme that was consistent with an active form of the MMP-9. Plasminogen activator zymographs of the conditioned media from the cancer cells showed that CSF treatment resulted in an increase in a 48-55 kDa plasminogen-dependent gelatinolytic activity that was characterized as human uPA. The conditioned medium from the cancer cells treated with CSFs stimulated the conversion of plasminogen to plasmin, providing a direct demonstration of the ability of enhanced uPA to increase plasmin-dependent proteolysis. The enhanced invasive behaviour of the cancer cells stimulated by CSFs was well correlated with the increase in MMPs and uPA activities. These data suggest that the enhanced production of extracellular matrix-degrading proteinases by the cancer cells in response to CSF treatment may represent a biochemical mechanism which promotes the invasive behaviour of the cancer cells.

Adenocarcinoma↗
PubMed1999

IL-4 inhibits the migration of human Langerhans cells through the downregulation of TNF receptor II expression.

The migration of Langerhans cells is an initial event in the sensitization phase of contact sensitivity. Langerhans cells travel from the epidermis to the regional lymph node, and can be variously modulated in the skin where many cytokines are released from epidermal cells, dermal cells, T helper (Th) cells, and other inflammatory cells during the sensitization and elicitation phase of contact dermatitis, and thus induce an altered inflammatory skin reaction. The modulatory effect of the cytokines released in the skin, such as IL-1beta, GM-CSF, and TNF-alpha as epidermal cytokines, IL-2, IL-12, and IFN-gamma as Th1 type cytokines, and IL-4 and IL-10 as Th2 type cytokines, was analyzed using the chemotactic chamber method in this study. Both GM-CSF and TNF-alpha induced the migration of human Langerhans cells in vitro, whereas IL-1beta, IL-2, IL-10, IL-12, and IFN-gamma had no effect on Langerhans cell migration. In contrast, IL-4 inhibited Langerhans cell migration in a dose dependent manner. The inhibitory activity of IL-4 was reversed by both anti-human IL-4 monoclonal antibody and anti-human IL-4 receptor monoclonal antibody. IL-4 inhibited the Langerhans cell migration induced by both TNF-alpha and GM-CSF. Furthermore, anti-TNF-RII monoclonal antibody inhibited both random migration and the migration induced by TNF-alpha, but not that induced by GM-CSF. A reverse-transcriptase-polymerase chain reaction and fluorescence-activated cell sorter analysis revealed that TNF-alpha up-regulated and IL-4 downregulated the TNF receptor II (TNF-RII) expression of Langerhans cells at both the mRNA and the protein levels. The pretreatment of Langerhans cells with TNF-alpha enhanced the migration of Langerhans cells and the expression of TNF-RII. After pretreating Langerhans cells with TNF-alpha, IL-4 inhibited both the migration of Langerhans cells and the expression of TNF-RII in a time dependent manner. These results indicate that IL-4 inhibits the migratory activity of Langerhans cells by downregulating the expression of TNF-RII in human Langerhans cells and thereby modulates the immune response in the skin.

Antibodies, Monoclonal↗
PubMed1999

Thiomenthol derivatives as novel percutaneous absorption enhancers.

Thiomenthol derivatives were synthesized and their promoting activity on the percutaneous absorption of ketoprofen from hydrogels was evaluated in rats. The apparent penetration rate (Rp) of ketoprofen absorbed from the hydrogel was estimated by the pharmacokinetic model derived under the assumption of a constant penetration rate through the skin after a lag time. As an index of promoting activity of thiomenthol derivatives, an enhancement factor (Ef) was defined as the ratio of the Rp value with enhancer to the value obtained with the control not containing enhancer. Skin irritancy evoked by these derivatives was investigated microscopically by using a cross-section of the excised skin onto which ketoprofen hydrogel was applied. Total irritation score (TIS) was estimated by summation of each irritation score in several parts of the skin. The physicochemical parameters of thiomenthol derivatives such as a partition coefficient (log P) and a steric energy were calculated and the quantitative relationships between these parameters and the Ef values or TIS values were investigated on the basis of multiple regression analysis. As a result, a parabolic relationship between log P and Ef was noted. A similar relationship was also observed in the case of TIS.

Administration, Cutaneous↗
PubMed1999

Encapsulation of hydrophilic and lipophilic drugs in PLGA nanoparticles by the nanoprecipitation method.

The purpose of this study was to assess the relative advantages and drawbacks of the nanoprecipitation-solvent displacement method for a range of drugs with respect to the particle size and drug encapsulation in polylactic-co-glycolic acid (PLGA) nanoparticles. The particle size analysis indicated a unimodal particle size distribution in all systems, with a mean diameter of 160-170 nm, except for insulin nanoparticles, which showed a smaller particle size. The results of the encapsulation efficiency analysis demonstrated that more lipophilic drugs, such as cyclosporin and indomethacin, do not suffer from the problems of drug leakage to the external medium, resulting in improved drug content in the nanoparticles. In spite of the fact that valproic acid is a liquid that is very sparingly soluble in water, very low encapsulation efficiency was obtained. Ketoprofen, a drug sparingly soluble in water, demonstrated intermediate values of encapsulation that were well correlated with its intermediate lipophilicity. More hydrophilic drugs, such as vancomycin and phenobarbital, were poorly encapsulated in PLGA nanoparticles. Insulin was preferentially surface bound on the PLGA nanoparticles. However, a strong hypoglycemic effect of the insulin was observed after administration of the suspension of PLGA nanoparticles with surface-bound insulin to the ileum loop of male Wistar rats.

Anesthesia↗
PubMed1999

The relationship between the rigidity of the liposomal membrane and the absorption of insulin after nasal administration of liposomes modified with an enhancer containing insulin in rabbits.

The relationship between the rigidity of the liposomal membrane and the absorption of insulin after nasal administration of liposomes modified with an enhancer containing insulin was investigated for the nasal delivery of peptide drugs in rabbits. The rigid liposomal membrane makes liposomes stable, protecting insulin from enzymatic degradation. Soybean-derived sterol (SS) or its sterylglucoside (SG) was used as an enhancer. Dipalmitoylphosphatidylcholine (DPPC) liposomes modified with SG had increased fluidity of the hydrophobic group of the liposome bilayer compared with the liposomes modified with cholesterol (Ch) or SS, as shown by measurements of the steady-state fluorescence anisotropy of 1,6-diphenyl-1,3,5,-hexatriene (DPH); however, the fluidity of the polar group of the liposome bilayer was decreased according to measurements of steady-state fluorescence anisotropy of dansylhexadecylamine (DSHA) at 37 degrees C. These findings suggest that the fluidity of the hydrophobic group of the liposome bilayer is responsible for the increase of liposomal leakage and instability of the liposomes. When insulin was administered nasally to rabbits as a solution, no hypoglycemic effect was observed. The administration of insulin contained in DPPC/SG (7/4, mole) liposomes with high fluidity caused a high glucose reduction of long duration (8 hr). DPPC/SS and DPPC/Ch (7/4) liposomes with low fluidity caused low glucose reductions. These results demonstrated that liposomes modified with SG can be useful as carriers of insulin administered nasally.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed1999

Second-line chemotherapy with weekly cisplatin and irinotecan in patients with refractory lung cancer.

Cisplatin and irinotecan are reported to act synergistically. The authors have conducted a phase II trial combining cisplatin and irinotecan in patients with refractory lung cancer to evaluate the activity and safety of the regimen. Twenty-one patients, who had not responded to prior platinum-based chemotherapy, were entered into the study. Both cisplatin (30 mg/m2) and irinotecan (60 mg/m2) were administrated on days 1, 8, and 15, and the regimen was repeated every 28 days. There were six partial responses, and the response rate was 29% (95% confidence interval, 11.3%-52.2%). The median survival time of all patients was 32 weeks, and 1-year and 2-year survival rates were 43% and 11%, respectively. Major toxicities were hematologic; leukopenia of grades 3 and 4 developed in 43% patients, anemia developed in 38%, and thrombocytopenia developed in 19%. One notable nonhematologic toxicity was diarrhea; which was grade 3 in 38%. The weekly chemotherapy combining cisplatin and irinotecan was active against lung cancer, which is refractory to platinum-based chemotherapy. However, skips of drug administration or dose reduction were necessary in 76% patients during two courses of planned administration, though the ratio of actual dose to scheduled dose was 81%. Dose modification would be necessary to yield better results by this weekly chemotherapy.

Adult↗
PubMed1999

Application of extracorporeal shock wave on bone: preliminary report.

BACKGROUND: We have studied the effect of extracorporeal shock waves (ESW) on bone. ESW emitted by the new powerful generator provides three to six times greater energy than a common lithotriptor. Because the ESW causes fracture of rabbit femurs and induces new bone formation, we have called this treatment as ESWIB (ESW-induced bone formation). The purpose of this study is to confirm the effect of ESWIB on a canine model, which is more similar to clinical cases, and to apply ESWIB on nonunion of clinical cases. METHODS: In our basic research, ESWIB was applied on six canine femurs as follows: group I with 100, 500, 1,000 shots and group II with 100, 500, 1,000 shots. A femur was extracted immediately after ESWIB in group I and 2 months after ESWIB in group II. Blood tests, including blood cell counts and blood chemistry studies, were performed before and after ESWIB in group II. In our clinical research, we applied ESWIB to six patients of delayed or nonunion of the fracture. The sites of the ESWIB application were three tibiae, one radius, one femur, and one humerus. Average age of the patients, the period from the previous surgery, and the period until fusion was achieved were 38.6 years, 14.0 months, and 4.3 months, respectively. RESULTS: In our basic research, group I, 500 or more shots caused periosteum detachment. In addition, small fractures of the inner surface of the cortex were observed. However, gross fracture with displacement was not observed. In group II, 500 or more shots caused callus formation beneath the detached periosteum. Subcutaneous hemorrhage was seen in all dogs, and the degree of bleeding was directly proportional to the number of the shots. The blood was absorbed within a week. The level of serum creatine kinase was significantly high 2 days after ESWIB, but it recovered in a week. In our clinical research, four of the patients achieved union without any complications except mild subcutaneous bleeding. CONCLUSIONS: We predict that ESWIB will be one of the tactics for treatment of the delayed union or nonunion of the fracture in the future.

Adult↗
PubMed1999

Expression of p53 and glutathione S-transferase-pi relates to clinical drug resistance in non-small cell lung cancer.

To determine the predictive value of the expression of p53 and glutathione S-transferase-pi (GST-pi) with respect to chemotherapy response, immunostaining was performed on transbronchial biopsy specimens from previously untreated patients with non-small cell lung cancer. Of the 54 patients, 34 patients (63%) and 37 patients (69%) were positive for p53 and GST-pi, respectively. The response rates in the p53-positive and p53-negative group were 15 and 45%, and those in GST-pi-positive and GST-pi-negative groups were 16 and 47%, respectively. A multiple logistic regression analysis revealed that positive immunostaining for GST-pi was a significant risk factor for clinical chemotherapy resistance. The combination of these two markers was the most important independent factor in predicting a response to chemotherapy in multiple logistic regression analysis. Immunohistochemical expression of p53 and GST-pi was independently related to clinical chemoresistance in patients with non-small cell lung cancer. Combined use of these two biomarkers may be a useful predictor of clinical chemoresistance.

Adult↗
PubMed1999

Frequent expression of p53 protein without mutation in the atypical epithelium of human bronchus.

We investigated the correlation between p53 protein levels and mutations in the p53 gene of atypical bronchial epithelium (ABE). Protein levels were analyzed by immunohistochemistry, whereas mutations were assessed by polymerase chain reaction-single-strand conformational polymorphism (PCR-SSCP) and direct sequencing. A total of 78 formalin-fixed, paraffin-embedded bronchial biopsy specimens that had been diagnosed to be ABE were retrieved from the archives and examined. p53 protein was expressed in 44 of the 78 (56%) specimens overall. However, when pathologically classified, 38% of hyperplasias, 58% of metaplasias, and 73% of dysplasias were positive, indicating that an increased frequency of p53 expression correlated with the severity of ABE (P = 0.042). Among the 44 specimens that expressed p53 protein, 40 (91%) did not reveal mutations by PCR-SSCP. In the four specimens with abnormal PCR-SSCP bands, p53 gene mutation was identified by direct sequencing and revealed the same point mutation at codon 248 (CGG-to-CTG transversion) of exon 7 in all four specimens. These four specimens were dysplasias derived from patients with lung cancer. p53 protein expression in ABE was associated with the wild-type gene in most cases; therefore, wild-type p53 protein expressed in ABE might have a protective function from lung carcinogenesis, and mutation of p53 gene may be a late event in the sequential steps of lung carcinogenesis.

Adolescent↗