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Biomedical subjects

T Handa

Publications and source records attributed to T Handa.

At least 91 records · Page 5Linked to original sources

Role of antibodies against outer-membrane proteins in murine resistance to infection with encapsulated Klebsiella pneumoniae.

In the assessment of immunity to the encapsulated virulent strain of Klebsiella pneumoniae and its avirulent mutant defective for capsular polysaccharide (CPS), killed bacterial vaccine of both strains could protect mice equally against challenge with 100 x LD50 of encapsulated wild strain. Antisera to each strain conferred the same level of protection on naive mice upon transfer; the protective anti-mutant serum was highly capable of opsonizing the encapsulated bacteria. In addition to the common antigenic components shared by both strains, the wild strain had antigen(s) unrelated to the mutant since the protective capacity of the anti-wild serum was not affected by preabsorption with the mutant strain; the protection conferred by the anti-mutant serum was mediated by antibodies against non-capsular antigens since the antiserum did not contain antibodies against purified CPS detectable by ELISA. As possible candidates among the non-capsular antigens, outer-membrane proteins (OMPs) extracted from the mutant strain were examined for their immunogenicity. Immunoblotting of the protein-containing fraction and ELISA using LPS-free OMP suggested that a number of proteins were involved in the immune response evoked by K. pneumoniae. Furthermore, mice immunized with OMP or anti-OMP serum could overcome a lethal challenge with the wild strain. These results indicated that OMPs of K. pneumoniae are implicated as the protective antigens and may pave the way for the development of non-capsular, proteinaceous vaccines.

Animals↗

Preparation of prolonged-release spherical micro-matrix of ibuprofen with acrylic polymer by the emulsion-solvent diffusion method for improving bioavailability.

Prolonged-release spherical micro-matrices of ibuprofen with acrylic polymer (Eudragit RS) were prepared using a novel emulsion-solvent diffusion method. It was found by examining cross sections of the spherical matrix before and after dissolution tests with a scanning electron microscope and a porosimeter that the resultant micro-matrix had a sponge-like internal structure. The spherical matrices were successfully recovered with a relatively high concentration of the drug in ethanol (0.4-0.6 g/ml) and over a wide range of temperatures (5-35 degrees C). The size of the spherical matrix could be easily controlled by varying the agitation speed of the system and the concentration of emulsifier added to the aqueous medium. The drug release rate from the spherical matrix decreased with increasing concentration of polymer formulated due to the reduced diffusion path and increased tortuosity in the matrix. Spherical matrices with ibuprofen:Eudragit RS = 3:1 improved the bioavailability of the drug and prolonged the drug action in beagle dogs.

Acrylates↗

In vitro primary induction of T cells mediating delayed footpad reaction and acquired cellular resistance to Listeria monocytogenes.

We established an in vitro system generating L. monocytogenes-specific T cells primarily from unprimed spleen cells of mice. Normal spleen cells were cultured for 5 days in the presence of L. monocytogenes in vitro. Viable cells were harvested and assessed for their capacity to confer acquired cellular resistance (ACR) and delayed footpad reaction (DFR) upon local passive transfer to naive syngeneic recipient mice. When normal spleen cells were stimulated with viable L. monocytogenes, the viable cells that were recovered after 5 days of culture conferred a high level of ACR and DFR. Negative selection revealed that the effector cells obtained in primary in vitro culture were Thy 1+, L3T4+, Lyt2- cells. T cells mediating ACR could not be generated in the culture of normal spleen cells with heat-killed bacteria; however, cells mediating only DFR were generated in the presence of a large number of killed L. monocytogenes. The expression of DFR and ACR by T cells generated in this primary culture system was Listeria-specific; reactions were not observed against unrelated bacterial antigens including S. typhimurium, S. aureus, E. coli and PPD. FACS analysis of the cells in culture showed that L3T4+ and Lyt2- T cells were being enriched during culture. The primary generation of antigen-specific T cells in vitro was also possible with spleen cells from NTx mice but not with cells from nude mice, suggesting the presence of Listeria-specific precursors in NTx mice.

Animals↗

The relationship between walking speed and muscle strength for knee extension in hemiparetic stroke patients: a follow-up study.

Walking speed and isokinetic strength of knee extension were examined in 18 hemiparetic stroke patients prior to, and 4 and 8 weeks after gait training. Although increase of the walking speed did not coincide with that of the isokinetic strength of the affected and non-affected sides, regression analyses revealed that the strength of the affected side was the primary determinant of the walking speed and the variance explained by it gradually increased with a period of the training.

Aged↗

Walking cycle after stroke.

Temporal-distance parameters of gait, maximum walking speed, walking rate and stride length, were successively examined in 10 hemiparetic stroke patients for 8 weeks after starting gait training. In patients with walking speed less than 20 m/min i.e., walking rate less than 90 steps/min, the increase of walking speed was significantly related to that of walking rate and stride length. In patients with walking speed greater than 20 m/min, the increase of walking speed correlated only to that of stride length, indicating that a limiting factor of walking speed in hemiparetic patients was the decreased stride length.

Adult↗

Effect of ankle-foot orthosis (AFO) on body sway and walking capacity of hemiparetic stroke patients.

Body sway, the total length of the sway of the center of foot pressure (CFP) and maximum walking speed were examined with and without AFO in eight post-stroke hemiparetic patients. Without AFO, the CFP moved towards the non-affected limb and the body sway was large. Wearing AFO, the CFP shifted to the midposition and the body sway became small. Without AFO, the time to walk the prescribed distance was longer, the cadence slower and the steps shorter than with AFO. However, there was no correlation between the improvements in body sway and walking capacity. The AFO compensated only for the instability of the ankle joint but not for the dysfunction of the central nervous system after the stroke.

Aged↗

Localization of dural fistulas using metrizamide digital subtraction fluoroscopic cisternography.

Demonstration of the exact site of dural fistulas in cases of cerebrospinal fluid rhinorrhea is difficult. Previous reports have described the use of metrizamide cisternography combined with either hypocycloidal tomography or computerized tomography; however, direct, dynamic, real-time visualization of the fistula is difficult with instillation of a minimal dose of metrizamide using those methods. A digital video subtraction fluoroscopy system can visualize the actual site of the fistula directly and dynamically using only a small amount of metrizamide.

Adult↗

Co-operative effect of MCF and MAF(IFN-gamma) in the protection of mice against Listeria monocytogenes.

The effects of macrophage chemotactic factor (MCF) and macrophage-activating factor (MAF) on protection to Listeria monocytogenes were analysed using 'MCF-rich fraction' and murine recombinant interferon-gamma (rIFN-gamma). Recombinant IFN-gamma showed no macrophage chemotactic activity in the assays performed in vitro and in vivo. Although a single injection of either MCF-rich fraction or rIFN-gamma into the footpads of mice led to a significant degree of anti-listerial activity, the highest degree of protection was observed when injected together. The i.v. administration of 20,000 U rIFN-gamma did not raise significant protective activity against Listeria; however, injection with rIFN-gamma prior to that with the MCF-rich fraction into the footpad produced a higher level of protective activity than the group treated with MCF alone. In addition to MAF(IFN-gamma), MCF seems to play another important role in the full expression of protection against L. monocytogenes.

Animals↗

Protective immunity to Listeria monocytogenes in neonatally thymectomized (NTx) mice: involvement of T cells distinct from those in sham-thymectomized mice.

Neonatally thymectomized (NTx) mice, whose ability to mount antigen-specific cell-mediated immunity is reported to be generally defective, were found to be capable of mounting a normal level of acquired cellular resistance (ACR) and delayed footpad reaction (DFR) to Listeria monocytogenes. The present study was done in order to determine the functional differences of T cells contributing to the protection against L. monocytogenes between NTx and sham-operated mice. In mice immunized with viable L. monocytogenes, the absolute number of splenic T cells was significantly lower in NTx mice compared with sham-operated mice. When the ability of immune T cells to transfer ACR and DFR was examined by passive transfer, lymphocytes from immune NTx mice conferred a higher level of ACR and DFR on naive recipient mice, despite the marked difference in total number of T cells compared with immune Sham mice. Antigen-specific proliferation and interleukin-2 (IL-2) production by splenic T cells from immune NTx mice were significantly lower than in those from immune Sham mice. The proliferative response of T cells to exogenous IL-2 was also lower in NTx group. These results suggest that the requirement for the IL-2-driven T-cell proliferation system is basically low in the generation of effector T cells specific for L. monocytogenes.

Animals↗

Particle design of tolbutamide in the presence of soluble polymer or surfactant by the spherical crystallization technique: improvement of dissolution rate.

Poorly soluble crystals of tolbutamide were modified in the presence of a soluble polymer or surfactant by the spherical crystallization technique, the objective being to improve the dissolution rate and to transform platelet crystals into spherical agglomerates. An HCI solution was added to a tolbutamide:NaOH solution containing a water-soluble polymer or surfactant. The tolbutamide crystals were agglomerated with either and were free flowing and spherically compact. The size of the crystals of the agglomerate depended on the viscosity of the solvent and adsorption of the surfactant onto the crystal surface. The tolbutamide-agglomerated crystals dissolved isotropically, with no evidence of disintegration. The dissolution process was described in terms of the Hixson-Crowell equation. The dissolution rate of the agglomerate was 8 times faster than that of conventionally crystallized tolbutamide. Therefore, the solubility and flow-ability of tolbutamide can be improved using the spherical crystallization technique.

Crystallization↗

A significant role of the macrophage accumulation induced by MCF in the protection of mice against Listeria monocytogenes in vivo.

Analysis was done on macrophage chemotactic factor (MCF) produced in the culture supernatant of spleen cells from mice immunized with Listeria monocytogenes. MCF was produced by Thy-1+, Lyt-1+ lymphocytes. MCF activity was resistant against pH 2 treatment and heating at 56 degrees C for 30 min, but was abolished by digestion with trypsin. G-100 gel filtration chromatography revealed that the approximate molecular weight of MCF was 15,000. MCF-rich fraction obtained by gel filtration chromatography showed neither MAF activity nor interferon activity. MCF activity in MCF-rich fraction was not affected by treatment with anti-rIFN-gamma antibody. An injection of MCF-rich fraction into the peritoneal cavity of mice induced a significant degree of accumulation of polymorphonuclear leukocytes (PMN) in a very short time after injection and macrophages thereafter. Resistance against listerial infection was augmented at the site where macrophage accumulation was provoked by the injection with MCF-rich fraction. It was shown that MCF plays an important role by itself in the protection against listerial infection by the accelerated accumulation of macrophages.

Animals↗

Enhancement of the dissolution rate of a poorly water-soluble drug (tolbutamide) by a spray-drying solvent deposition method and disintegrants.

The dissolution rate of a poorly water-soluble drug, tolbutamide, was improved by spray-drying a diluted ammonia solution of the drug containing either a low-substituted hydroxypropylcellulose (L-HPC) or partly pregelatinized corn starch (PCS) as disintegrants. With L-HPC the resultant particles were agglomerates of disintegrant with drug on the surface and within the particles, while particles formed with PCS were composed of a single core of PCS on which the drug was deposited. The deposited drug crystals were very fine because the rapid solvent evaporation restricted crystal growth. The spray-dried particles prepared with PCS had a structure similar to that of an ordered mix. The drug dissolution rate from the spray-dried particles was more rapid than that of the powdered drug alone or with disintegrant and could be attributed to separation of the layer of fine drug crystals from the surface of the particles by swelling of disintegrant. PCS enhanced the drug dissolution rate compared with systems using corn starch. The dissolution rate also depended on the drug content of the particles which was higher than that in ordered mixtures or conventional solvent deposition systems. This system described also had the advantage of avoiding the use of organic solvents.

Chemical Phenomena↗

Gamma interferon-mediated increase in the number of Ia-bearing macrophages during infection with Listeria monocytogenes.

The role of gamma interferon (IFN-gamma) in an increase in Ia-bearing macrophages during Listeria monocytogenes infection was studied. The peritoneal macrophages from L. monocytogenes-infected mice contained a high proportion of Ia. Intraperitoneal injection of the supernatant from a culture of spleen cells from L. monocytogenes-infected mice induced Ia-rich exudates in normal mice. The Ia-inducing activity in the culture supernatant was abrogated by the pretreatment of spleen cells with anti-Thy-1.2 antibody plus complement. Immunoadsorption of the culture supernatant with anti-recombinant IFN-gamma antibody and protein A-Sepharose CL-4B completely abrogated its Ia-inducing activity. These results suggested that an increase in Ia-bearing macrophages during L. monocytogenes infection was attributable to T-cell-derived IFN-gamma.

Animals↗

Induction by killed Listeria monocytogenes of effector T cells mediating delayed-type hypersensitivity but not protection in mice.

Using a local passive transfer system, we found that effector T cells mediating delayed-type hypersensitivity (DTH) but not acquired cellular resistance (ACR) to Listeria monocytogenes (strain EGD) were generated in mice immunized with killed Listeria, although immunized mice did not express DTH or ACR. When non-adherent cells of peritoneal, lymph node, or spleen cells from mice immunized with killed Listeria were transferred into the footpad of naive recipient mice along with eliciting antigen, positive delayed footpad reaction (DFR) was elicited. However, there was no evident protection against challenge at the site of the local transfer. Cells from mice immunized with viable Listeria conferred significant degrees of DFR and ACR on the recipients. DFR transferred by cells immunized with killed Listeria was mediated by L3T4+ T cells in an antigen-specific manner. The antigen-specific proliferative response of T cells from mice immunized with killed Listeria was much lower than that of T cells from mice immunized with viable Listeria. The production of macrophage chemotactic factor (MCF) by cells from killed Listeria-immune mice was much the same as that by cells from viable Listeria-immune mice. In contrast, the production of interleukin-2 (IL-2) and macrophage activating factor (MAF) was much lower in cells from killed Listeria-immune mice. The elimination of L. monocytogenes (strain L461), a strain of low virulence, was enhanced at the site of DFR transferred with cells from killed Listeria-immune mice. These results suggest that stimulation with killed bacteria is effective for the generation of DTH-mediating effector T cells, and that different effector T cells mediating DTH or ACR are involved in cell-mediated immunity to L. monocytogenes.

Animals↗

[The diagnostic method of localization of cerebrospinal fluid rhinorrhea by digital video subtraction system].

Demonstration of the exact site of the dural fistula in cerebrospinal fluid rhinorrhea is difficult. Previous reports present the methods of metrizamide cisternography combined with both hypocycloid tomography and computed tomography. But in these methods, direct, dynamic, actual and real-time visualization of the fistula with minimal dose of metrizamide is rather difficult. By using digital video subtraction system (Philips DVI-2CV), we could visualize the direct, dynamic and actual site of fistula with small amount of metrizamide instilled into the suboccipital subarachnoid space with the patient prone position. We report a successful case of traumatic cerebrospinal fluid rhinorrhea drained through the bony defect at the planum sphenoid into the posterior ethmoid sinus. This is the first report to deal with the usefulness of digital video subtraction system for exact localization of cerebrospinal fluid rhinorrhea. We emphasize the usefulness of metrizamide cisternography by the digital video subtraction system combined with the metrizamide computed tomographic cisternography for the precise localization of dural fistula.

Adult↗

Effects of lysophosphatidylcholine micelle and phosphatidylcholine liposome on photoreduction of methylviologen.

Photoinduced reduction of methylviologen (MV2+) by ethylenediaminetetraacetate (EDTA3-), which was sensitized by thiacarbocyanine dyes having long alkyl chains (C+m-n) embedded in palmitoyl lysophosphatidylcholine micelle and dipalmitoyl phosphatidylcholine liposomal membrane, was carried out. The formation rate of reduced methylviologen cation radical (MV+.) decreased with the time of irradiation with visible light, and the deceleration was more pronounced in the micellar solution. In kinetic studies, we found that the sensitizer divalent cation radical (C2+.m-n) is formed through the reaction of photoexcited sensitizer (C+*m-n) with MV2+ as an intermediate in this reaction, and that the reduction of C2+.m-n with EDTA3- inhibits the back reaction of MV+. with C2+.m-n. The inhibition was greater in the liposomal solution than in the micellar solution. This was ascribed to a higher concentration of EDTA3- on the liposomal surfaces through the electrostatic interaction between EDTA3- and the liposomal surfaces, the charge of which is attributed to the univalent cation sensitizer embedded in the liposomal membrane. The difference in the positive charge density of the surface of these lipid aggregates was due to the difference in the curvature of the micelle and the liposome. These results suggest that the dipalmitoyl phosphatidylcholine liposome is a more effective carrier than the palmitoyl lysophosphatidylcholine micelle for the production of MV+. in the photoreduction studied here.

Edetic Acid↗