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Biomedical subjects

S Kameda

Publications and source records attributed to S Kameda.

At least 19 recordsLinked to original sources

Kidney-targeted naked DNA transfer by retrograde injection into the renal vein in mice.

We recently developed a novel kidney-targeted gene transfer technique in rats, using the retrograde renal vein injection of naked plasmid DNA. Many animal disease models are created in mice by transgenic or knockout technologies. However, it is much harder to perform renal vein injection in mice than in rats because they have a thin and short vein. Here we transferred the mouse interleukin (IL)-10 gene into mice by retrograde renal vein injection, using an IL-10 and immunoglobulin fusion protein (IL-10/Fc) (96-kDa) expression plasmid, pCAGGS-IL10/Fc. We observed a dose-response relationship between serum IL-10 levels and the amount of injected DNA. The serum IL-10 levels peaked at day 1 and then were sustained for at least 2 weeks. These results demonstrate that the kidney-targeted naked plasmid DNA transfer of mice by retrograde renal vein injection can be achieved, and the kidney serves as a depot organ for the production of large proteins.

Animals↗

Post-secretion neutralization of transgene-derived effect: soluble erythropoietin receptor/IgG1Fc expressed in liver neutralizes erythropoietin produced in muscle.

BACKGROUND: The regulation of transgene expression is a key issue for the development of safe gene therapy. Various strategies have been used to regulate protein production at the levels of transgene expression, transcription, translation, and secretion. Neutralization following secretion is another important backup system to prevent super-therapeutic levels of a protein from being expressed by gene transfer. METHODS: We tested whether the soluble human erythropoietin receptor (EpoR)/IgG(1)Fc could neutralize the rat Epo at the post-secretory level and suppress erythrocytosis. RESULTS: To assess whether soluble human EpoR could bind rat Epo in vitro, we used the Epo-dependent human leukemic cell line, AS-E2. EpoR/IgG(1)Fc significantly inhibited the growth of AS-E2 cells in Epo-containing medium. To test this neutralization effect of EpoR/IgG(1)Fc in vivo, we first transferred pCAGGS-Epo into rat muscle by in vivo electroporation, confirmed erythropoiesis for 3 weeks, and then delivered EpoR/IgG(1)Fc by liver-targeted gene transfer via tail-vein injection with hydrodynamics-based transfection. Reticulocyte counts and hematocrit levels in rats that received pCAGGS-EpoR/IgG(1)Fc injections were significantly lower than in rats that received pCAGGS-EpoR, pCAGGS-IgG(1)Fc, or no injection. CONCLUSIONS: These results demonstrate that liver-targeted pCAGGS-EpoR/IgG(1)Fc transfer by tail-vein injection with hydrodynamics-based transfection is useful for neutralizing Epo delivered by in vivo electroporation. This backup strategy at the level of post-secretion could facilitate the clinical application of gene therapy in the future.

Animals↗

Hydrodynamics-based transfer of PCR-amplified DNA fragments into rat liver.

A high level of plasmid DNA expression in rat liver can be achieved by the rapid injection of a large volume of a naked DNA solution into the tail vein, called the 'hydrodynamics-based procedure.' The preparation of PCR-amplified DNA fragments is easier than that of naked DNA. In this paper we evaluated the effects of expressing the erythropoietin (Epo) gene in the rat liver by injecting fCAGGS-Epo, an Epo-expressing PCR-amplified DNA fragment, via the tail vein. After injection of 5 pmol fCAGGS-Epo (10 microg) or pCAGGS-Epo (18.4 microg), plasmid DNA, the serum Epo levels peaked at week 1, then persisted for at least 12 weeks. Transgene-derived Epo secretion resulted in significant erythropoiesis. These results demonstrated that transfer of PCR-amplified DNA fragments into the rat liver via rapid tail vein injection can be achieved. This method may provide a useful means for studying the physiologic function of a putative gene.

Animals↗

Hydrodynamics-based delivery of the viral interleukin-10 gene suppresses experimental crescentic glomerulonephritis in Wistar-Kyoto rats.

Gene therapy is expected to revolutionize the treatment of kidney diseases. Viral interleukin (vIL)-10 has a variety of immunomodulatory properties. We examined the applicability of vIL-10 gene transfer to the treatment of rats with crescentic glomerulonephritis, a T helper 1 (Th 1) predominant disease. To produce the disease, Wistar-Kyoto rats were injected with a rabbit polyclonal anti-rat glomerular basement membrane antibody. After 3 h, a large volume of plasmid DNA expressing vIL-10 (pCAGGS-vIL-10) solution was rapidly injected into the tail vein. pCAGGS solution was similarly injected into control rats (pCAGGS rats). We confirmed the presence of vector-derived vIL-10 mainly in the liver and observed high serum vIL-10 levels in pCAGGS-vIL-10-injected rats. Compared with the pCAGGS rats, the pCAGGS-vIL-10 rats showed significant therapeutic effects: reduced frequency of crescent formation, decrease in the number of total cells, macrophages, and CD4+ T cells in the glomeruli, decrease in urine protein, and attenuation of kidney dysfunction. Using quantitative real-time polymerase chain reaction, we also observed that this model was Th1-predominant in the glomeruli and that the ratio of the transcripts of CD4, interferon-gamma, tumor necrosis factor-alpha, and monocyte chemotactic protein-1 to the transcripts of glucose-6-phosphate dehydrogenase in the glomeruli were all significantly lower in the pCAGGS-vIL-10 rats than in the pCAGGS rats. These results demonstrate that pCAGGS-vIL-10 gene transfer by hydrodynamics-based transfection suppresses crescentic glomerulonephritis.

Animals↗

Acute eosinophilic pneumonia caused by cigarette smoking.

It has been suggested that acute eosinophilic pneumonia (AEP) is associated with cigarette smoking because in Japan, the patients with AEP are young and have a high incidence of short-term smoking history. However, there has been no direct evidence to support that cigarette smoke causes AEP. Herein is reported the first case showing the direct evidence and a long-term clinical course of cigarette smoking-induced AEP, in which tolerance to repeated resumption of smoking cigarettes might have occurred. We should pay attention to the history of cigarette smoking in seeing patients with AEP, especially in young patients.

Adult↗

Real time image processing with an analog vision chip system.

A linear analog network model is proposed to characterize the function of the outer retinal circuit in terms of the standard regularization theory. Inspired by the function and the architecture of the model, a vision chip has been designed using analog CMOS Very Large Scale Integrated circuit technology. In the chip, sample/hold amplifier circuits are incorporated to compensate for statistic transistor mismatches. Accordingly, extremely low noise outputs were obtained from the chip. Using the chip and a zero-crossing detector, edges of given images were effectively extracted in indoor illumination.

Adaptation, Ocular↗

Clinical study of the pumping pressure changes in the temporomandibular joint space before and after arthroscopic surgery.

When arthroscopy of the temporomandibular joint (TMJ) is performed, we often observe differences in the resistance of hydraulic pressure during intra-articular pumping. It can be speculated that intra-articular adhesion and contracture of the capsule and ligament may cause these differences in resistance. In order to measure the changes in intra-articular hydraulic pressure before and after arthroscopic ablation surgery, seven cases were examined by balloon pumping technique. We concluded that the values of intra-articular hydraulic pressure are implicated in the severity of the internal derangement of TMJ.

Angioplasty, Balloon↗

Mouse trophoblast cell invasion of extracellular matrix purified from endometrial tissue: a model for peri-implantation development.

We have investigated the invasive activity of mouse trophoblast cells during embryo implantation in vitro by culturing blastocysts with extracellular matrix (ECM) purified from mouse endometrium obtained on day 4 of pregnancy. Endometrium was dissected from lyophilized mouse uteri, and intact ECM was isolated by sequential precipitation in nonionic detergent and high salt. Electron microscopic examination of the ECM revealed typical collagen fibers plus an amorphous material resembling basement membrane. Electrophoretic analysis of the ECM revealed an enrichment of high molecular weight proteins, and immunoblotting indicated the presence of fibronectin, laminin, entactin, and type IV collagen, but not the intracellular proteins 2',3'-cyclic nucleotide-3'-phosphodiesterase or vimentin. Mouse blastocysts cultured with this ECM attached to it within 3 days, and the trophoblast cells began to migrate through the matrix in a manner resembling trophoblast invasion in utero. Unlike blastocysts cultured on plastic surfaces, the trophoblast did not flatten and become disorganized, but retained a polarized, spherical structure. Fluorescent microscopy with fluorescein isothiocyanate-labeled phalloidin revealed a high degree of microfilament organization and established that actin was absent from the ECM preparation. In the presence of a serum substitute, differentiation continued through yolk sac formation. Without serum components, yolk sac did not form; however, light and electron microscopic examination indicated that the invasive behavior of trophoblast cells persisted and was comparable to that of trophoblasts cultured in the presence of the serum substitute. A three-dimensional model for investigating trophoblast behavior in ECM from the endometrium should be of great value in elucidating the cellular and molecular events surrounding the process of blastocyst implantation.

Animals↗

Terminal patterns of the tegmental afferents in the interpeduncular nucleus: a Golgi study in the mouse.

The intranuclear course, distribution and termination of the tegmental afferents in the interpeduncular nucleus (IP) were studied in the mouse by means of the rapid Golgi method. Primarily on the basis of terminal branching patterns and distribution areas, two types of afferents were distinguished. The type 1 fibers distribute mainly within the rostral half in the form of numerous glomerular endings, the size of which corresponds well with that of the tufted terminal dendrites of the IP neurons. On the other hand, the caudal half of the IP has far fewer fibers than the rostral IP and is innervated by the type 2 fibers, which follow a tortuous course, terminating in dense fiber plexus. Thus, the rostral and caudal IP are innervated in a different fashion by different afferents originating from tegmental regions. These results are discussed in relation to the distribution patterns of another conspicuous afferent system of the IP, the fasciculus retroflexus.

Afferent Pathways↗

Terminal patterns of the fasciculus retroflexus in the interpeduncular nucleus of the mouse: a Golgi study.

The courses and terminal patterns of the fasciculus retroflexus (FR) in the interpeduncular nucleus (IP) were studied in the mouse, using the rapid Golgi method. Mainly on the basis of the distribution areas and terminal patterns, the FR fibers are divided into two types. The type 1 FR fibers are coarse in contour and take zigzag courses to distribute throughout the entire rostral half and core region of the caudal IP. In contrast, the type 2 fibers are fine, travel caudally along the lateral boundary of the IP and terminate in the lateral regions of the caudal half, forming a dense fiber plexus. The distribution areas of the type 1 and type 2 fibers are clearly differentiated from each other, from the cytoarchitectural as well as the fibroarchitectural viewpoint. Thus, the type 1 and type 2 FR fibers form different fiber systems in the IP. These results are discussed in the light of the known hodological, histochemical and ultrastructural studies.

Animals↗

A Golgi study on the olfactory bulb in the red stingray, Dasyatis akajei.

The intrinsic organization of the olfactory bulb (OB) was studied in the red stingray using the rapid Golgi method. The OB is horse shoe-shaped, surrounding the equator region of the nasal capsule. As seen in the sagittal sections, the OB is round with the long olfactory peduncle extending from the dorsocaudal region and the olfactory fibers in a thick bundle entering from the rostroventral aspect. Although not so distinct, the following areas are distinguished. A rostroventral ovoid area adjacent to the entrance of the olfactory fibers consists exclusively of the olfactory fibers running in various directions. Dorsocaudal to the olfactory fiber area is a wide crescent region containing thin bundles of olfactory fibers, olfactory glomeruli, mitral cells and a few disseminated granule cells. A narrow crescent area made up of scattered granule cells is located dorsocaudally to the above wide crescent area. The outermost region consists of a fiber layer encapsulating the dorsal to caudal aspect of the OB. Thus, while the major constituents of the vertebrate OB are recognized, the lamination is very obscure.

Animals↗

[A case of tsutsugamushi disease in the urban area of Komatsu City].

Most of patients with tsutsugamushi disease are diagnosed by their clinical histories suggesting the opportunities of Rickettsia infection in a rural region. We reported a 76-year-old female patient, who was considered to be infected in her house in the urban area of Komatsu City. She has shown typical clinical manifestations of tsutsugamushi disease, and was remitted successfully by oral administration of minocycline. Although specific antibodies to Rickettsia tsutsugamushi could not be detected in her serum by the complement fixation (CF) method during her clinical course, their significant elevation was confirmed by the indirect immunofluorescence (IF) method.

Aged↗

A Golgi study on the neuronal organization of the habenular ganglion in the red stingray, Dasyatis akajei.

The neuronal organization of the habenular ganglion (HG) was studied in the red stingray using the rapid Golgi method. The HG was made up of the medial (MH) and lateral habenular nucleus (LH), and the former nucleus was further divided into a dorsal, intermediate and ventral subnucleus. Only one type of neurons were observed in the MH, while the LH was composed of two types of neurons. In the left HG cut at the rostrocaudal middle of the ganglion, the LH was located in the dorsolateral region, while the dorsal, intermediate and ventral subnuclei of the MH occupied the dorsomedial, intermediate and ventral portions of the ganglion, respectively. In contrast, the right ganglion seen at this level was composed exclusively of the MH, with the dorsal, intermediate and ventral subnuclei located in the dorsomedial, intermediate and ventral portions, respectively. In the caudal level of the left ganglion, each nucleus was seen almost in the same region as in the level of the rostrocaudal middle, however, three subnuclei of the MH fused with the same subnuclei of the opposite side. In the right ganglion at the caudal level, the LH appeared in the intermediate area. The right LH was far smaller and was located more ventrocaudally than the left LH. On account of the LH, the intermediate subnucleus of the MH was divided into a dorsal and ventral part. The dorsal and ventral subnuclei of the MH remained in the same region as in the rostral level. Thus, the HG of the red stingray exhibited a striking left-right asymmetry, the most remarkable aspect of which was considered to be differences of the size, form and location of the LH between the left and right HG.

Animals↗

A Golgi study on the afferent fibers to the habenular ganglion in the red stingray, Dasyatis akajei.

Afferent fibers to the habenular ganglion (HG) were derived mainly from the stria medullaris thalami (SM), which was roughly divided into a dorsal and ventral bundle. In the left ganglion seen at the level of the rostrocaudal middle, the dorsal bundle gave off collaterals to the lateral habenular nucleus (LH) and dorsal subnucleus of the medial habenular nucleus (MH), while the ventral bundle innervated the intermediate and ventral subnuclei of the MH. On the other hand, in the right ganglion at the level of the rostrocaudal middle, the dorsal subnucleus of the MH was innervated by collaterals from the dorsal bundle of the SM, whereas in the intermediate and ventral subnucleus fibers from the ventral bundle were seen. In the left ganglion at the caudal level, the dorsal and ventral bundle extended medially and joined the same bundle of the opposite side to constitute a dorsal and intermediate component of the habenular commissure, respectively. A third component of the HC, a ventral component, was seen to run between the fasciculus retroflexus of both sides. As in the case of the rostral level, the dorsal bundle of the SM emitted collaterals to the LH and dorsal subnucleus of the MH, while the intermediate and ventral subnuclei of the MH were projected upon by collaterals from the ventral bundle of the SM. At the caudal level of the right ganglion, the dorsal bundle gave off collaterals to the dorsal subnucleus of the MH. In contrast, the LH and the intermediate and ventral subnuclei of the MH were innervated by fibers from the ventral bundle. With regard to terminal patterns of the SM, fibers to the MH gave off many short fine branchlets forming the glomerular structures, whereas those to the LH branched out into numerous terminals to form a dense fiber plexus. Thus, the afferent fibers to the HG in the red stingray exhibited a striking left-right asymmetry.

Afferent Pathways↗

Sheehan's syndrome with hypomagnesemia and polymorphous ventricular tachycardia.

A 40 year old woman with Sheehan's syndrome demonstrated polymorphous ventricular tachycardia and hypomagnesemia. The diagnosis of Sheehan's syndrome was based on her history, physical examination and endocrinological data. Serum magnesium level was 1.5 mg/dl. After the treatment of polymorphous ventricular tachycardia by lidocaine, an electrocardiogram showed low voltage, T-wave inversion or flattening and QT-prolongation, and these electrocardiographic abnormalities gradually improved after the initiation of cortisone therapy. A chest roentgenogram, an echocardiogram, thallium-201 scan of the myocardium and hemodynamic parameters indicated no abnormal findings. It is suspected that hypomagnesemia caused polymorphous ventricular tachycardia. However, the cause of hypomagnesemia and its relation to Sheehan's syndrome in the present case were not determined, although, hypomagnesemia was improved after the administration of cortisone.

Adult↗