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Biomedical subjects

S Kuroda

Publications and source records attributed to S Kuroda.

At least 163 records · Page 9Linked to original sources

Purification, characterization, and sequence determination of phospholipase D secreted by Streptoverticillium cinnamoneum.

Phospholipase D (PLD), secreted into the culture medium of an actinomycete, Streptoverticillium cinnamoneum, has been purified to homogeneity and characterized. The Stv. cinnamoneum PLD efficiently catalyzes both the hydrolysis and transphosphatidylation of various phospholipids, including phosphatidylethanolamine (PE), phosphatidylcholine (PC), and phosphatidylserine (PS). However, the substrate specificity differs between the two reactions; PE serves as the most preferred substrate for the hydrolysis, but PC and PS are better substrates than PE for the transphosphatidylation. In addition, the transphosphatidylation but not the hydrolysis of PE and PC is markedly activated on the addition of metal ions, especially Al3+. Nucleotide and amino acid sequence determination of the Stv. cinnamoneum PLD revealed the presence of common structural motifs identified in all PLD sequences from various species.

Aluminum↗

Neuroprotective effects of a novel nitrone, NXY-059, after transient focal cerebral ischemia in the rat.

Recent results have demonstrated that the spin trapping agent alpha-phenyl-N-tert-butyl nitrone (PBN) reduces infarct volume in rats subjected to 2 hours of middle cerebral artery occlusion, even when given 1 to 3 hours after the start of recirculation. In the current study, the authors assessed the effect of NXY-059, a novel nitrone that is more soluble than PBN. Loading doses were given of 0.30, 3.0, or 30 mg x kg(-1) followed by 0.30, 3.0, or 30 mg x kg(-1) x h(-1) for 24 or 48 hours. Dose-response studies showed that when treatment was begun 1 hour after recirculation, 0.30 mg x kg(-1) had a small and 30 mg x kg(-1) a marked effect on infarct volume. At equimolar doses (3.0 mg x kg(-1) for NXY-059 and 1.4 mg x kg(-1) for PBN), NXY-059 was more efficacious than PBN. Similar results were obtained when a recovery period of 7 days was allowed. The window of therapeutic opportunity for NXY-059 was 3 to 6 hours after the start of recirculation. Studies of the transfer constant of [14C]NXY-059 showed that, in contrast to PBN, this more soluble nitrone penetrates the blood-brain barrier less extensively. This fact, and the pronounced antiischemic effect of NXY-059, suggest that the delayed events leading to infarction may be influenced by reactions occurring at the blood-endothelial interface.

Animals↗

Insulin-induced phosphorylation and activation of cyclic nucleotide phosphodiesterase 3B by the serine-threonine kinase Akt.

Cyclic nucleotide phosphodiesterase (PDE) is an important regulator of the cellular concentrations of the second messengers cyclic AMP (cAMP) and cGMP. Insulin activates the 3B isoform of PDE in adipocytes in a phosphoinositide 3-kinase-dependent manner; however, downstream effectors that mediate signaling to PDE3B remain unknown. Insulin-induced phosphorylation and activation of endogenous or recombinant PDE3B in 3T3-L1 adipocytes have now been shown to be inhibited by a dominant-negative mutant of the serine-threonine kinase Akt, suggesting that Akt is necessary for insulin-induced phosphorylation and activation of PDE3B. Serine-273 of mouse PDE3B is located within a motif (RXRXXS) that is preferentially phosphorylated by Akt. A mutant PDE3B in which serine-273 was replaced by alanine was not phosphorylated either in response to insulin in intact cells or by purified Akt in vitro. In contrast, PDE3B mutants in which alanine was substituted for either serine-296 or serine-421, each of which lies within a sequence (RRXS) preferentially phosphorylated by cAMP-dependent protein kinase, were phosphorylated by Akt in vitro or in response to insulin in intact cells. Moreover, the serine-273 mutant of PDE3B was not activated by insulin when expressed in adipocytes. These results suggest that PDE3B is a physiological substrate of Akt and that Akt-mediated phosphorylation of PDE3B on serine-273 is important for insulin-induced activation of PDE3B.

3',5'-Cyclic-AMP Phosphodiesterases↗

Regulation of the cytoskeleton and cell adhesion by the Rho family GTPases in mammalian cells.

Members of the Rho family of small Ras-like GTPases--including RhoA, -B, and -C, Rac1 and -2, and Cdc42--exhibit guanine nucleotide-binding activity and function as molecular switches, cycling between an inactive GDP-bound state and an active GTP-bound state. The Rho family GTPases participate in regulation of the actin cytoskeleton and cell adhesion through specific targets. Identification and characterization of these targets have begun to clarify how the Rho family GTPases act to regulate cytoskeletal structure and cell-cell and cell-substratum contacts in mammalian cells. The Rho family GTPases are also involved in regulation of smooth muscle contraction, cell morphology, cell motility, neurite retraction, and cytokinesis. However, the molecular mechanisms by which the Rho family GTPases participate in the regulation of such processes are not well established.

Cell Adhesion↗

Cell adhesion and Rho small GTPases.

The Rho small GTPases, Cdc42, Rac1 and Rho, are implicated in regulation of integrin-mediated cell-substratum adhesion and cadherin-mediated cell-cell adhesion. Identification and characterization of effectors of these GTPases have provided insights into their modes of action. Rho-kinase, an effector of Rho, regulates integrin-mediated cell-substratum adhesion (focal adhesion) by regulating the phosphorylation state of myosin light chain (MLC): it directly phosphorylates MLC and also inactivates myosin phosphatase. IQGAP1, an effector of Cdc42 and Rac1, regulates cadherin-mediated cell-cell adhesion by interacting with (beta)-catenin and dissociating (alpha)-catenin from the cadherin-catenins complex. Activated Cdc42 and Rac1 inhibit IQGAP1, thereby stabilizing the cadherin-catenins complex. Cdc42/Rac1 and IQGAP1 thus appear to constitute a switch that regulates cadherin-mediated cell-cell adhesion.

Cadherins↗

Human NELL-1 expressed in unilateral coronal synostosis.

Surgical correction of unilateral coronal synostosis offers a unique opportunity to examine the molecular differences between an abnormal and a normal cranial suture. We isolated and identified a cDNA fragment whose expression was up-regulated in the premature fusing and fused coronal sutures, as compared with normal coronal sutures. The nucleotide sequence of the full-length cDNA of this gene, human NELL-1, has approximately 61% homology with the chicken Nel gene. Both chicken Nel and human NELL-1 are comprised of six epidermal growth factor-like repeats. The human NELL-1 messages were localized primarily in the mesenchymal cells and osteoblasts at the osteogenic front, along the parasutural bone margins, and within the condensing mesenchymal cells of newly formed bone in sites of premature sutural fusion. Human multiorgan tissue mRNA blot showed that NELL-1 was specifically expressed in fetal brain but not in fetal kidney, liver, or lung. We also showed that Nell-1 was expressed in rat calvarial osteoprogenitor cells and was largely absent in rat tibiae and fibroblast cell cultures. In conclusion, our data suggest that the NELL-1 gene is preferentially expressed in cranial intramembranous bone and neural tissue (both of neural crest cell origin) and is up-regulated during unilateral premature closure of the coronal suture. The precise role of this gene is unknown.

Amino Acid Sequence↗

Depression in the early stages of Pick's disease.

To better understand the nature of the symptoms of depression in the early stages of Pick's disease, we performed a retrospective study of the medical records of eight patients who were originally treated for major depressive disorders before being clinically diagnosed with Pick's disease. Six of the eight manifested psychomotor retardation and social withdrawal, seven of the eight were agitated and five of the eight showed hyperbulia too. However, only two of the eight showed melancholia or physical symptoms such as insomnia or loss of appetite. All patients were treated with antidepressants but these were not effective in relieving the symptoms of depression. The data we gathered in this study will be useful in the future for distinguishing between Pick's disease-related depression (in the early stages of the disease) and major depression.

Affect↗

Delirium in elderly patients admitted to a psychiatric hospital: causative factors and treatment.

We studied the causes and treatments of delirium in 43 patients admitted to the geriatric-psychiatric ward of a hospital in Japan. Patients studied were divided into three groups according to the type of factor responsible for their delirium. We termed these causal factors precipitating, facilitating and predisposing factors. Twenty-one patients exhibited precipitating factors, the most common of which were overmedication and poisoning. Almost all these cases were treated with psychotropic drugs. Facilitating factors were judged responsible in nine cases. In six of these, admission to a hospital was thought to be the facilitating factor. Thirteen patients who had only predisposing factors were treated with psychotropic drugs while 27% of patients without precipitating factors were successfully treated for delirium without the use of such drugs.

Aged↗

Anorexia nervosa with severe liver dysfunction and subsequent critical complications.

A twenty-year-old woman with anorexia nervosa (body mass index=11) suffered from severe liver dysfunction (aspartate aminotransferase 5,000 IU/l, alanine aminotransferase 3,980 IU/l, prothrombin time 32%), hypoglycemia (serum glucose 27 mg/dl), and pancreatic dysfunction (amylase 820 IU/l, lipase 558 IU/l). She fell into a depressive state with irritability, which was not improved by intravenous glucose. Despite treatment with plasmapheresis for the liver dysfunction, she subsequently developed pulmonary edema, acute renal failure, gastrointestinal bleeding, and disseminated intravascular coagulation. Hemodialysis, mechanical ventilation and drug therapy including prednisolone, prostaglandin E1, and branched-chain amino acid, improved her critical condition. In this case, malnutrition may have been the cause for the liver dysfunction and subsequent complications.

Acute Kidney Injury↗

Telemedicine in neurosurgery using international digital telephone services between Japan and Malaysia--technical note.

A new image transmission and teleconference system using international digital telephone services was established between Japan and Malaysia. This new system consists of an ordinary personal computer, image scanner, and terminal adapter for digital telephone lines. The quality of images transferred using this system was high enough for diagnosis and discussion except for images such as radiographs requiring huge data transfer. Transmission of one image took approximately 20 seconds. The cost performance was almost equal to the conventional mailing system. The most remarkable advantage of this new system is the high quality of transferred images, the cost and time performance, and security of the medical information. New communication systems using international digital networks including the internet may allow re-distribution of medical resources between advanced countries and developing countries in neurosurgery.

Academic Medical Centers↗

Long-term patency of radial artery graft bypass for reconstruction of the internal carotid artery. Technical note.

Reconstruction of the carotid artery by using a radial artery graft is a useful option that can produce reliable long-term patency for the surgical treatment of giant and/or large aneurysms of the cavernous and paraclinoid internal carotid artery (ICA). During the past 10 years, 43 patients with intracavernous and paraclinoid giant aneurysms of the ICA have been treated by reconstruction of the ICA with radial artery grafts after ligation of the cervical ICA. The long-term patency of the grafted radial artery was evaluated over more than a 5-year period (mean 7.2 years) in 20 of these patients by using magnetic resonance angiography or conventional angiography. There was no late occlusion of the graft in any of these cases. Stenotic graft changes were observed in two cases.

Adult↗

[Development of a multiple side-hole sheath introducer for use in aortography].

We developed a multiple side-holed sheath introducer that can also be used as a catheter for aortography before selective arteriography of the pelvic organs or lower extremities, as an alternative to straight or pig-tail catheters, which were formerly used for aortography. Aortography using the multiple side holed sheath introducer provided good detection of arteries in 100% of 10 cases, without any complications or difficulties. Thus, the present sheath introducer can replace straight or pig-tail catheters, resulting in the more economical use of one catheter prior to selective arteriography.

Aortography↗

[Digital recording in neurosurgery].

The purpose of this study is to examine the image quality of multi-media for digital recording of surgical procedure using an operating microscope in neurosurgery. For video recording, high vision recording, digital video recording and analogue VHS recording were used. For still pictures, an analogue camera with 35 mm silver halide film and a micro-digital camera were used. The quality of photographs taken with a microdigital camera was superior to the quality of those taken with the conventional 35 mm film camera. The micro-digital camera system is superior to the conventional 35 mm camera in neurosurgery in its quality and success rate. In video recording, high vision analogue recording was superior to any other media as far as its image quality is concerned although its practical convenience is limited and cost performance is not always good. On the other hand, digital video can record high quality images, including still pictures, with satisfactory quality to the neurosurgeon. These digital recording media are also space saving for storing the huge amount of data obtained during surgery and the cost-performance is superior to that of the conventional method. In the near future, most of surgical procedures are supposed to be expected using digital media.

Humans↗

[Two cases of sporadic juvenile Parkinson's disease caused by homozygous deletion of Parkin gene].

Parkin gene, which is one of the causes of familial Parkinson's disease, was cloned in 1998, and it was found that mutations of this gene induces familial Parkinson's disease of autosomal recessive form (AR-JP). Clinical studies have revealed that almost cases of AR-JP are consanguineous or hereditary. However, AR-JP may also be caused by marriage between two carriers. We therefore investigated the existence of a homozygous deletion in the parkin gene in 10 patients with juvenile Parkinson's disease with the onset of age younger than 40 years without consanguinity or heredity. A large delection of the gene was found in 2 of the 10 patients with sporadic Parkinson's disease, and both of these deletion were exon 4. The present study clearly suggests that juvenile Parkinson's disease in Japan may be caused by abnormalities in parkin gene regardless of family history or consanguinity.

Age of Onset↗

A behavioral and immunohistochemical study on the development of perirhinal cortical kindling: a comparison with other types of limbic kindling.

We have previously reported that the perirhinal cortex (PRC) plays an important role in the generalization of kindled seizures. In the present study, we kindled the rat PRC and made a comparison with amygdala (AM) and dorsal hippocampal (dHIPP) kindling. In order to produce a functional map of the seizure generalization pathway from these limbic foci, we also stained for Fos protein in sections of the PRC-, AM- and dHIPP-kindled brains using an immunohistochemistry technique. In the generalized seizures of PRC kindling the duration of afterdischarges (ADs) and the latency to forelimb clonus were significantly shorter than those of AM kindling or dHIPP kindling. Typically, the PRC-kindled rats demonstrated moving arrest or exploratory behavior for about 10 days, and then a characteristic 'rapid backward moving' behavior for 1 day, followed by the sudden appearance of generalized motor seizures. Fos protein induction after a single stimulation of the PRC is more widely observed than after a single stimulation of the AM, in that the PRC stimulation produced Fos protein expression in the temporal and parietal neocortices. Following AM and PRC kindling, the Fos-positive areas were asymmetrically propagated from the ipsilateral to the contralateral hemisphere. The contralateral PRC was primarily activated at the generalization of epileptic activity in the contralateral hemisphere. In contrast, the Fos protein distribution of the dHIPP-kindled rats was restricted to the bilateral hippocampi during the early stages, followed by the symmetrical propagation from the limbic system to the neocortex during the generalized seizures. These results indicate that the PRC plays a characteristic role in the seizure generalization of kindling.

Amygdala↗

Posttranscriptional control of adipocyte differentiation through activation of phosphoinositide 3-kinase.

Differentiation of adipocytes is an important aspect of energy homeostasis. Although the transcriptional regulation of adipocyte differentiation is relatively well characterized, the subsequent molecular events remain unclear. The activity of phosphoinositide (PI) 3-kinase precipitated with antibodies to phosphotyrosine has now been shown to increase transiently during adipocyte differentiation of 3T3-F442A and of 3T3-L1 cells. PI 3-kinase activity precipitated with antibodies to insulin receptor substrate 1 (IRS1) and association of subunits of PI 3-kinase with IRS1 were also increased at this stage of differentiation, suggesting that IRS1 contributes to PI 3-kinase activation. Inhibition of the activation of PI 3-kinase by expression of dominant negative mutant subunits of the enzyme prevented adipogenesis, as assessed by lipid accumulation and expression of key adipocyte proteins such as GLUT4, adipsin, and aP2, suggesting that PI 3-kinase activation is essential for adipocyte differentiation. However, these mutant proteins did not affect either the expression of the transcription factor PPARgamma at the mRNA or protein level or the increase in the abundance of mRNAs encoding the adipocyte marker proteins. These results demonstrate that adipocyte differentiation is regulated at the posttranscriptional level and that activation of PI 3-kinase is required for this regulation.

3T3 Cells↗

Requirement of GM2 ganglioside activator for phospholipase D activation.

Sequence analysis of a heat-stable protein necessary for the activation of ADP ribosylation factor-dependent phospholipase D (PLD) reveals that this protein has a structure highly homologous to the previously known GM2 ganglioside activator whose deficiency results in the AB-variant of GM2 gangliosidosis. The heat-stable activator protein indeed has the capacity to enhance enzymatic conversion of GM2 to GM3 ganglioside that is catalyzed by beta-hexosaminidase A. Inversely, GM2 ganglioside activator purified separately from tissues as described earlier [Conzelmann, E. & Sandhoff, K. (1987) Methods Enzymol. 138, 792-815] stimulates ADP ribosylation factor-dependent PLD in a dose-dependent manner. At higher concentrations of ammonium sulfate, the PLD activator protein apparently substitutes for protein kinase C and phosphatidylinositol 4,5-bisphosphate, both of which are known as effective stimulators of the PLD reaction. The mechanism of action of the heat-stable PLD activator protein remains unknown.

Amino Acid Sequence↗