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Biomedical subjects

Daisuke Suzuki

Publications and source records attributed to Daisuke Suzuki.

At least 55 records · Page 3Linked to original sources
PubMed2004

Interaction of the spectrin-like repeats of alpha-actinin-4 with humanin peptide.

BACKGROUND: Podocyte alpha-actinin-4 (actinin-4) is an essential component of the glomerular filtration barrier. We recently reported that the central rod spectrin-like repeats (R1-R4) of actinin-4 have a high affinity to puromycin aminonucleoside (PAN), which can induce nephro-sis in animals. The aim of this study was to identify endogenous molecules that interact with the actinin-4 R1-R4 domain. METHODS: To identify such molecules, we performed a bacterial two-hybrid screening of a human kidney cDNA library using as a bait human actinin-4 R1-R4. We further verified the identified interactions by in vitro affinity assays and immunofluorescent studies of cultured human embryonic kidney HEK293 cells. To investigate the expression of the identified molecules in podocytes, in situ hybridization, and immunohistochemical studies were performed. RESULTS: One isolated cDNA from the library encoded humanin, a recently identified antiapoptotic peptide. In vitro affinity assays showed specific interactions of recombinant actinin-4 R1-R4, R1, R2, R3, and R4 proteins with humanin-Sepharose. PAN had no effect on these interactions. Green fluorescent protein-fused humanin and endogenous actinin colocalized mainly in the perinuclear cytoplasm of HEK293 cells. Altered colocalization was not observed by the addition of PAN. In situ hybridization and immunohistochemistry showed the expression of humanin in podocytes. CONCLUSIONS: Our results suggest that humanin is a novel binding partner of the actinin-4 R1-R4 domain in podocytes. Humanin and PAN are unlikely to compete for the same binding surface in actinin-4.

Actinin↗
PubMed2004

Comparative histology of lymph nodes from aged animals and humans with special reference to the proportional areas of the nodal cortex and sinus.

Lymph nodes are composed of a lymphocyte-rich area or cortex (subdivided into the superficial and deep cortex and the medullary cord) and another, macrophage-rich area (incorporating the subcapsular and medullary sinuses). We measured the proportional area of the cortex in lymph nodes from aged experimental mammals (rats, guinea pigs, dogs and rabbits) and elderly Japanese humans. The cervical, axillary and inguinal nodes were generally richer in cortex tissue than the pulmonary regional and mesenteric nodes. Histological heterogeneity and medullary sinus dominance were much more evident in the human nodes than in those from animals, except for the guinea pig thoracic node. Human pulmonary regional nodes were characterized by a large medullary sinus; in guinea pigs, these nodes had a similar histology but the T lymphocyte-containing areas were smaller and thinner than in humans. The paraaortic node was well developed in humans and dogs, but not in other animals tested. These species- and region-specific histological differences may influence the evaluation of experimental animal models of lymph node function, such as those recently identified for research into sentinel nodes.

Aged↗
PubMed2004

Expression of human nephrin mRNA in diabetic nephropathy.

BACKGROUND: Diabetic nephropathy (DN) is associated with functional changes in the filtration barrier, and microalbuminuria is a strong predictor of the development of overt DN. Nephrin is a novel podocyte-specific protein which localizes at the slit diaphragm. This study examines the expression of nephrin mRNA in the kidneys of type 2 diabetics with DN. METHODS: Renal tissues were obtained from 13 type 2 diabetics with DN. We also examined samples from five patients with minimal change nephrotic syndrome (MCNS) and five normal kidneys (normals) as control. The severity of DN was classified into two grades based on histopathological findings. DN grade 1 (DN1 = seven patients) presented mild mesangial expansion, and DN grade 2 (DN2 = six patients) moderate mesangial expansion. Nephrin mRNA was quantitated and localized by in situ hybridization. RESULTS: Cells positive for nephrin mRNA were detected exclusively in glomerular epithelial cells. The percentage of cells positive for nephrin mRNA in DN2 was significantly lower than in MCNS and normal kidneys. Furthermore, there was an inverse correlation between the percentage of cells positive for nephrin mRNA and extent of proteinuria. CONCLUSION: The low expression of nephrin mRNA may be closely linked to development and/or progression of proteinuria in human diabetic nephropathy.

Adult↗
PubMed2004

High expression of PKC-MAPK pathway mRNAs correlates with glomerular lesions in human diabetic nephropathy.

BACKGROUND: Activation of protein kinase C (PKC) is a major signaling pathway for transforming growth factor (TGF)-beta to induce extracellular matrix (ECM) production in diabetic nephropathy (DN). PKC also activates mitogen-activated protein kinase (MAPK), which is called the PKC-MAPK pathway. The PKC-MAPK pathway is probably responsible for PKC-related abnormalities in diabetic glomeruli. To confirm the involvement of this pathway, we determined the localization and expression of mRNAs in glomeruli by in situ hybridization method. METHODS: In the present study, we examined expression of PKCbeta1, MAPK/ERK kinase (MEK) 1, MEK2, extracellular signal-regulated protein kinase (ERK) 1, ERK2, and TGF-beta1 mRNAs using renal tissue samples from kidneys affected by DN (N= 21) and from normal human kidney (NHK; N= 6). We also performed an immunohistochemical study using anti-phosphorylated MEK1/2 (P-MEK) and ERK1/2 (P-ERK) antibodies. The glomerular severity of DN was classified into three groups according to mesangial expansion: D1 (N= 4), D2 (N= 13), and D3 (N= 4). We analyzed differences and correlations between variables. RESULTS: In the glomeruli, the number of cells that stained for these mRNAs in DN was significantly higher than in NHK. The expression of PKC-MAPK pathway mRNAs tended to be inversely proportional to the degree of mesangial expansion. The P-MEK and P-ERK signal intensity were parallel to its mRNA expression pattern. Furthermore, there were significant correlations among the P-MEK, P-ERK signal intensity, PKCbeta1 mRNA expression. CONCLUSION: Our results suggest that high expression of PKC-MAPK pathway mRNAs plays an important role in the development and/or progression of early tissue damage in DN.

Adult↗
PubMed2004

A spectrum of clinical manifestations caused by host immune responses against Epstein-Barr virus infections.

Epstein-Barr virus (EBV), or human herpesvirus 4 (HHV-4), infects the vast majority of adults worldwide, and establishes both nonproductive (latent) and productive (lytic) infections. Host immune responses directed against both the lytic and latent cycle-associated EBV antigens induce a diversity of clinical symptoms in patients with chronic active EBV infections who usually contain an oligoclonal pool of EBV-infected lymphocyte subsets in their blood. Episomal EBV genes in the latent infection utilize an array of evasion strategies from host immune responses: the minimized expression of EBV antigens targeted by host cytotoxic T lymphocytes (CTLs), the down-regulation of cell adhesion molecule expression, and the release of virokines to inhibit the host CTLs. The oncogenic role of latent EBV infection is not yet fully understood, but latent membrane proteins (LMPs) expressed during the latency cycle have essential biological properties leading to cellular gene expression and immortalization, and EBV-encoded gene products such as viral interleukin-10 (vIL-10) and bcl-2 homologue function to survive the EBV-infected cells. The subsequent oncogenic DNA damage may lead to the development of neoplasms. EBV-associated NK/T cell lymphoproliferative disorders are prevalent in Asia, but quite rare in Western countries. The genetic immunological background, therefore, is closely linked to the development of EBV-associated neoplasms.

Epstein-Barr Virus Infections↗
PubMed2003

Rectification efficiency of a Brownian motor.

The energy balance of a Brownian motor is discussed based on a Langevin equation without the overdamped approximation. Energetics of the system suggests that the frictional dissipation energy associated with the unidirectional movement should be counted as a part of the useful energy for the rectification process of a Brownian motor. This leads to a new definition of the efficiency, which is applicable, contrary to the conventional one, even if the external load is absent. For the so-called flashing ratchet model, we numerically solve the Langevin equation for various situations and discuss both the temperature and the friction strength dependence of the rectification efficiency and the role of the duty ratio.

Journal Article↗
PubMed2003

Increase of TCR V beta accessibility within E beta regulatory region influences its recombination frequency but not allelic exclusion.

Seventy percent of the murine TCRbeta locus (475 kb) was deleted to generate a large deleted TCRbeta (beta(LD)) allele to investigate a possible linkage between germline transcription, recombination frequency, and allelic exclusion of the TCR Vbeta genes. In these beta(LD/LD) mice, the TCRbeta gene locus contained only four Vbeta genes at the 5' side of the locus, and consequently, the Vbeta10 gene was located in the original Dbeta1-Jbeta1cluster within the Ebeta regulatory region. We showed that the frequency of recombination and expression of the Vbeta genes are strongly biased to Vbeta10 in these mutant mice even though the proximity of the other three 5'Vbeta genes was also greatly shortened toward the Dbeta-Jbeta cluster and the Ebeta enhancer. Accordingly, the germline transcription of the Vbeta10 gene in beta(LD/LD) mice was exceptionally enhanced in immature double negative thymocytes compared with that in wild-type mice. During double negative-to-double positive transition of thymocytes, the level of Vbeta10 germline transcription was prominently increased in beta(LD/LD) recombination activating gene 2-deficient mice receiving anti-CD3epsilon Ab in vivo. Interestingly, however, despite the increased accessibility of the Vbeta10 gene in terms of transcription, allelic exclusion of this Vbeta gene was strictly maintained in beta(LD/LD) mice. These results provide strong evidence that increase of Vbeta accessibility influences frequency but not allelic exclusion of the TCR Vbeta rearrangement if the Vbeta gene is located in the Ebeta regulatory region.

Alleles↗
PubMed2003

Enol ester as an olefinic partner in enyne cyclization. A novel tandem cyclization to stereodefined bicyclo[3.3.0]octenes.

When (Z)-1-tridecen-6-ynyl propanoate was treated with a titanium(II) alkoxide reagent, Ti(O-i-Pr)4/2i-PrMgCl, a novel tandem cyclization took place to give (1RS,2RS,3SR)-3-ethyl-4-hexyl-4-bicyclo[3.3.0]octene-2,3-diol virtually as a single stereoisomer after aqueous workup. The generality of this cyclization was also shown. Similarly, (Z)-3-butyl-1-tridecen-6-ynyl propanoate afforded (1RS,2RS,3SR,8SR)-8-butyl-3-ethyl-4-hexyl-4-bicyclo[3.3.0]octene-2,3-diol with a diastereoselectivity of >95:5, demonstrating the simultaneous construction of a total of four consecutive stereogenic centers in the product and with virtually complete diastereoselectivity.

Journal Article↗
PubMed2003

Crocodilian bone-tendon and bone-ligament interfaces.

We investigated bone-tendon (27 sites) and bone-ligament (12 sites) interfaces in six pairs of crocodile limbs and girdles under light microscopy. These crocodilian interfaces often included a direct, unmediated insertion in which the tendon or ligament fibers inserted directly into the bone itself without fibrocartilaginous mediation. This was quite different from the usual direct insertion known in mammals and lizards. Fibrocartilaginous tissue at the bone-tendon interface is generally believed to protect tendon fibers against shear stress. Other types of insertions were found in the crocodilian epiphyses, namely, hyaline cartilage and pseudofibrocartilaginous insertions. Notably, a thick periosteum/perichondrium and subchondral layer was involved at both interfaces. The thick periosteum/perichondrium seemed to form along the epiphyseal hyaline cartilage and might function in replacement of fibrocartilaginous tissues. Crocodilian thick periosteum/perichondrium would be expected to reinforce the limb and girdle bones--especially their epiphyses, in which secondary centers of ossification are absent. The subchondral layer--a kind of fibrocartilaginous tissue--seemed to play the role of the growth plate in compensating for the absence of secondary centers of ossification. Therefore, we hypothesized that the crocodile-specific bone-tendon interfaces were the result of these specializations of bone development and growth. In crocodiles, the disadvantages of the single ossification center are effectively compensated for by specialized morphologies, including these interfaces. Specialized bone growth provides the crocodile with the largest body size of the recent reptiles and an extremely fast method of locomotion.

Alligators and Crocodiles↗
PubMed2003

Glomerular expression of connective tissue growth factor mRNA in various renal diseases.

Connective tissue growth factor (CTGF) is a cysteine-rich member of a new family of growth regulators. It is an important factor in the pathogenesis of mesangial matrix accumulation and progressive glomerulosclerosis. The present study was designed to elucidate the role of CTGF in diabetic nephropathy (DN), immunoglobulin A nephropathy (IgA-N), membranous nephropathy (MN), and minimal change nephrotic syndrome (MCNS). We evaluated the expression and localization of CTGF mRNA in surgically excised renal tissue samples from 10 patients with DN, 10 with IgA-N, 10 with MN, 10 with MCNS, and 10 normal human kidney (NHK) tissue samples, by using high-resolution in situ hybridization with digoxigenin-labelled oligonucleotide. To quantify CTGF mRNA expression, we counted all nuclei, and nuclei surrounded by CTGF-positive cytoplasm, in at least 10 randomly selected cross-sections of non-sclerotic glomeruli, and expressed the results as a percentage of total glomerular cells. In all glomeruli, CTGF mRNA was expressed mainly in glomerular intrinsic cells, including glomerular mesangial and epithelial cells and some cells of Bowman's capsule. The percentage of cells positive for CTGF mRNA was significantly higher in DN and IgA-N than in MN, MCNS and NHK. However, there was no significant difference in the percentage of CTGF mRNA-positive cells between DN and IgA-N. Our study indicates that CTGF may play an important role in the development and progression of glomerulosclerosis in DN and IgA-N, which are both accompanied by mesangial matrix expansion and comprise two major causes of end-stage renal failure.

Adult↗
PubMed2002

Selective preparation of benzyltitanium compounds by the metalative Reppe reaction. Its application to the first synthesis of alcyopterosin A.

Dialkoxytitanacyclopentadienes, prepared from two different acetylenes and a divalent titanium alkoxide reagent, Ti(O-i-Pr)4/2 i-PrMgCl, reacted with propargyl bromide to give directly benzyltitanium compounds. The resultant benzyltitanium compounds underwent deuteriolysis, iodinolysis (with I2), or oxygenation (with O2 gas) to give the corresponding deuterium-labeled compounds, iodides, or alcohols, illustrating their synthetic versatility. The first synthesis of alcyopterosin A, a bicyclic aromatic sesquiterpenoid recently isolated and characterized, has been achieved by this method, starting with an appropriate combination of an acetylene and a diyne.

Benzyl Compounds↗
PubMed2002

Selective syntheses of metalated pyridines from two different unsymmetrical acetylenes, a nitrile, and a titanium(II) alkoxide.

Cyclotrimerization of two different, unsymmetrical acetylenes and p-toluenesulfonylnitrile with a divalent titanium alkoxide reagent, Ti(O-i-Pr)4/2 i-PrMgCl, yielded single pyridyltitanium compounds in a highly selective manner. These metalated pyridines were confirmed by deuteriolysis to give the corresponding deuterated pyridines and underwent iodinolysis and copper-catalyzed alkylation to demonstrate their synthetic utility. Alternatively, a different type of cyclotrimerization of an alkynamide, terminal acetylenes, and alpha-alkoxynitriles mediated by the same titanium(II) alkoxide again proceeded in a highly selective manner to give single pyridines having a titanated side chain.

Journal Article↗
PubMed2002

Histology of the bone-tendon interfaces of limb muscles in lizards.

Lizards exhibit continual bone growth at the epiphysis, and their limb muscles are distributed differently from those of mammals because of differences in weight bearing. We therefore characterized the bone-tendon (B-T) interface (also termed the enthesis) in lizards. Using the forelimbs of five monitor lizards and three iguanas, we performed histological investigations on 57 B-T interfaces. Most reptilian tendons were very short and were often composed of wavy fiber bundles. Fibrocartilage (FC)-mediated direct insertion was observed at all epiphyses, whereas periosteum-mediated indirect insertions, including fleshy attachments, were often located on the flat surfaces of the pectoral girdles and at the diaphyses of the limb bones. The reptilian B-T interface was characterized by variability in the morphology of the FC-mediated insertions, especially by morphologies intermediate between those of FC- and periosteum-mediated interfaces; i.e., 1) various degrees of absence of the clear FC zonation seen in mammals, including the tidemark; 2) involvement of the periosteum in the FC; 3) the presence of various types of FC cells in the tendon near the interface, to reinforce the tendon against compression or shear stress; and 4) both FC and hyaline cartilage (lateral articular cartilage) receiving the tendon at the epiphysis. Overall, variations in the connective tissue, especially the FC tissue, were very evident in the reptilian B-T interface. The specific structures of the interfaces probably represent adaptations to the continuous growth and loose joint structures of lizards.

Animals↗
PubMed2002

Two-dimensional mapping of impedance magnetocardiograms.

A new method for measuring two-dimensional (2-D) impedance magnetocardiograms (I-MCGs) and magnetocardiograms (MCGs) above the heart simultaneously, has been developed. The I-MCG's and MCG's signals are recorded by using a superconducting interference device (SQUID) system. To measure the I-MCG and MCG signals, four first-order gradiometers with an 18-mm diameter and a 50-mm baseline were used. The SQUIDs are driven by a flux-locked-loop circuit with a frequency range higher than that of an ac-current (40 kHz) with constant amplitude passed through a subject. The output of the circuit is filtered through two circuits: one for measuring the I-MCG signals and one for measuring the MCG signals. The I-MCG signals are obtained by demodulating the magnetic field, which is detected by the gradiometers, at the frequency of the ac current. As a result, the I-MCG signal reflects the change in spatial distribution of conductivity caused by the movement of the heart muscle and blood volume. A contour map of the 2-D I-MCG signals showed the largest signals occur above the right ventricle and right atrium. In a corresponding current-arrow map, it was found that the large current arrows occurred above the right side of the right ventricle. Furthermore, it was found that the systole and diastole timings obtained from the first-derivative I-MCG signal and the phonocardiogram were different. These results show that primitive 2-D I-MCG signal can provide much physiological information on the circulatory movement of the heart.

Adult↗
PubMed2002

Renal proximal tubular metabolism of protein-linked pentosidine, an advanced glycation end product.

BACKGROUND: Pentosidine, an advanced glycation end product, accumulates in plasma proteins of uremic patients. Its fate is, however, yet to be fully understood. METHODS: Three cell lines, JTC-12 (proximal tubular cells), MDCK (distal tubular cells), and BALB3T3 (nonrenal cells), were cultured in a double chamber system and were exposed to uremic serum, and the contents of protein-linked pentosidine derived from uremic sera were determined in each medium by HPLC assay. The presence of pentosidine in the cytoplasm of these cells was assessed by immunoperoxidase staining. RESULTS: When the apical cell membrane was exposed to uremic serum (fortified in the upper chamber), the contents of protein-linked pentosidine in the upper medium decreased by up to 30% after 24- and 48-hour incubations of JTC-12 cells but not of other cells. On the other hand, the contents of protein-linked pentosidine in the lower medium did not change. By contrast, exposure of the basolateral cell membrane of the three cell lines to uremic serum (fortified in the lower chamber) did not change the contents of protein-linked pentosidine both in the upper and lower medium after a 24-hour incubation. Pentosidine was detected immunohistochemically in the cytoplasm of JTC-12 cells, but not of BALB3T3 and MDCK cells, the apical membranes of which were exposed to uremic sera for 8 h. The immunoreaction disappeared 48 h after exposure. Pentosidine was not detected in the cytoplasm of JTC-12 cells, the basolateral membranes of which were exposed to uremic sera. The relevance of the in vitro results to humans was demonstrated by immunohistochemical studies in normal human kidney tissues showing that pentosidine was identified in the proximal renal tubules. CONCLUSION: These results suggest that the proximal tubular cells play a role in the disposal of plasma pentosidine.

3T3 Cells↗
PubMed2002

Overexpression of the serpin megsin induces progressive mesangial cell proliferation and expansion.

Mesangial cells maintain normal glomerular function by mediating ECM remodeling and immune complex disposal. We have recently identified megsin, a novel member of the serine protease inhibitor (serpin) superfamily predominantly expressed in the mesangium. While our previous studies suggested a role for megsin in the pathogenesis of human glomerular diseases, its exact biological significance remained unknown. Here we produced two lines of megsin transgenic mice. Overexpression of megsin led to progressive mesangial matrix expansion and an increase in the number of mesangial cells. These glomerular lesions were accompanied by an augmented immune complex deposition, together with Ig's and complement. Binding and functional assays in vitro identified plasmin as one biological substrate of megsin and confirmed its activity as a proteinase inhibitor. Transgenic animals exhibiting nephritis as a result of treatment with anti--glomerular basement membrane antiserum showed significantly more persistent expansion of the mesangial ECM than was seen in parental mice. Megsin therefore exerts a biologically relevant influence on mesangial function, and on the mesangial microenvironment, such that simple overexpression of this endogenous serpin engenders elementary mesangial lesions.

Animals↗
PubMed2002

Direct raises in blood Ca levels by infusing a high-Ca solution into the blood stream accelerate the secretion of calcitonin from the ultimobranchial gland in eels.

In eels, a CaCl(2) solution was infused into the pneumatic duct vein. Plasma Ca levels were significantly increased during 3 hr and were followed by significant raises in plasma calcitonin levels. These results strongly suggest that, in eels, direct raises in blood Ca levels by infusion of a high-Ca solution via blood vessels can accelerate the secretion of calcitonin from the ultimobranchial gland.

Adaptation, Physiological↗