Publications by authors named "Chikako Murakami"

37 Publications

Individual identification using the RapidHIT™ ID system for forensic samples.

Leg Med (Tokyo) 2020 Nov 6;47:101776. Epub 2020 Aug 6.

Department of Legal Medicine, Kitasato University School of Medicine, 1-15-1 Kitasato, Minami-ku, Sagamihara, Kanagawa 252-0374, Japan.

The RapidHIT™ ID system produces GlobalFiler™ analysis results after a short operating time. This device is effective because it automatically extracts DNA from oral mucosal cells or from blood stains and saliva collected at a crime scene, with subsequent polymerase chain reaction performed to produce a DNA profile. Two types of dedicated cartridges are available for RapidHIT™ ID: the RapidHIT™ ID ACE GlobalFiler Express sample cartridge for oral cells and other samples and the RapidINTEL™ sample cartridge for minute samples, such as blood stains. Previously validated specimens include oral mucosa cells and blood stains left at crime scenes. There have been no reports of blood and nail clipping samples collected from the postmortem bodies at the time of death. This report summarizes the results of using the RapidHIT™ ID system by collecting a variety of actual forensic samples from postmortem bodies at different stages of decomposition, which were subsequently analyzed using these cartridges.
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http://dx.doi.org/10.1016/j.legalmed.2020.101776DOI Listing
November 2020

Extensive gastric necrosis secondary to acute gastric dilatation: A case report.

Leg Med (Tokyo) 2019 Feb 13;36:85-88. Epub 2018 Nov 13.

Department of Legal Medicine, Kitasato University School of Medicine, 1-15-1 Kitasato, Minami-ku, Sagamihara, Kanagawa 252-0374, Japan.

We report a case of sudden death in a patient who developed extensive gastric necrosis secondary to acute gastric dilatation. A 36-year-old man with mental retardation (but without difficulties in activities of daily living), developed an illness after a meal out with friends, necessitating 3 hospital visits. He returned home after receiving drug therapy; however, his condition deteriorated, and he was transferred to our hospital via ambulance. Whole-body computed tomography performed upon admission revealed gastric dilatation. A stomach tube was inserted, and 2000 mL of gastric aspirate was obtained. The patient died approximately 5 h later despite receiving treatment. Autopsy revealed 1000 mL of gastric contents and extensive gastric necrosis. He was diagnosed with extensive gastric necrosis secondary to acute gastric dilatation.
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http://dx.doi.org/10.1016/j.legalmed.2018.11.007DOI Listing
February 2019

Vulvar reconstruction should be performed using gluteal-fold perforator flap because of less morbidities and complications.

Rev Col Bras Cir 2014 Mar-Apr;41(2):134-6

Clinical Research Center, National Hospital Organization Nagasaki Medical Center, Nagasaki, Japan.

The authors present a case of bilateral vulvar defects after abrasion of malignant skin neoplasm, reconstructed with a gluteal-fold perforator flap, resulting in a successful outcome.
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http://dx.doi.org/10.1590/s0100-69912014000200011DOI Listing
March 2016

Emergent free flow-through anterolateral thigh flaps for Gustilo-Anderson III fracture of the upper extremity.

J Emerg Trauma Shock 2014 Jan;7(1):53-5

Department of Plastic and Reconstructive Surgery, Clinical Research Center, National Hospital Organization, Nagasaki Medical Center, Nagasaki, Japan.

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http://dx.doi.org/10.4103/0974-2700.125642DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3912654PMC
January 2014

Cutaneous malignant rhabdoid tumor in the palm of an adult.

Rare Tumors 2013 12;5(3):e36. Epub 2013 Jul 12.

Department of Plastic and Reconstructive Surgery, Clinical Research Center and National Organization, Nagasaki Medical Center , Nagasaki.

Malignant rhabdoid tumor is a rare tumor occurring mostly in the neonatal kidneys and central nervous system. Cutaneous malignant rhabdoid tumors are extremely rare in adults. The aim of the study was to report on the clinical, histologic, and immunophenotypic characteristics of this cutaneous malignant rhabdoid tumor which developed in an adult. A 27-year-old male complained of a right palm neoplasm that had been present for 6 months, which was initially diagnosed as an epithelioid sarcoma by biopsy. However, detailed investigation with immunohistochemistry enabled us to make a diagnosis of a rhabdoid tumor. The patient underwent radical abrasion, chemotherapy, and irradiation, and has survived for 1 year without relapse. Only 20 adult cases have been reported thus far in the English literature. We are reporting the 21(st) case, who remains disease-free at 12 months. Complete resection and local irradiation may increase survival, because there is no standard and reliable curative chemotherapeutic regimen.
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http://dx.doi.org/10.4081/rt.2013.e36DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3804811PMC
November 2013

Vascularized bone graft is a better option for the reconstruction of maxillary defects.

Eur Arch Otorhinolaryngol 2013 Sep 4;270(10):2779-81. Epub 2013 Jul 4.

We read the article of Kinnunen et al., which evaluated the result of maxillary defects, and feel some objections. We present our considerations of their operative indication and thoughts based on our surgical experiences. Defects after palatectomy, which have left no dentition for the retention of an obturator, require vascularized bone-containing free flaps. Local flaps are available in only small defects of Class 1 and 2a. Most palatomaxillary defects following malignant tumor abrasion are classified as 2b, 2c, 3, or 4, which require microsurgical free flap transfer combined with bony reconstruction. Regarding bony reconstruction, non-vascularized bone grafts tend to be absorbed. Thus, we believe that bony reconstruction should be performed with vascularized bone. We agree with the authors' comment that PTMF may be useful in repairing defects due to complications in microvascular procedures in the palatal area. However, even when bone segment is required for salvage surgery, using a vascularized bone flap is more preferable. A parietal bone-fascial-periosteal flap based on the superficial temporal vessels is a suitable and reliable bone flap for the reconstruction of a maxillary defect following free skin flap transfer to the palate.
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http://dx.doi.org/10.1007/s00405-013-2619-5DOI Listing
September 2013

Simultaneous determination for oxicam non-steroidal anti-inflammatory drugs in human serum by liquid chromatography-tandem mass spectrometry.

Forensic Sci Int 2013 Apr 27;227(1-3):100-2. Epub 2012 Dec 27.

Department of Pathophysiology, Yokohama College of Pharmacy, Yokohama, Kanagawa 245-0066, Japan.

A high-performance liquid chromatography-tandem mass spectrometry (LC/MS/MS) technique was developed for the simultaneous determination of five non-steroidal anti-inflammatory oxicam drugs (ampiroxicam, tenoxicam, piroxicam, meloxicam and lornoxicam) in human plasma. These five oxicam drugs and isoxicam (internal standard) were extracted from human plasma with an Oasis(®) MAX cartridge column and analysed on a Unison UK-C18 column (2.0 mm × 100 mm, 3 μm) with an acetonitrile:10mM formic ammonium buffer (pH 3.0) (50:50) mobile phase at 0.20 ml/min at 37°C. The analytes were detected using a tandem mass spectrometer, equipped with an electrospray ion source (ESI). The instrument was used in multiple-reaction-monitoring (MRM) mode. The extraction yields from a 200 μl human plasma sample (containing 10 ng of each drugs) with the Oasis(®) MAX cartridge column were 93.3-102.5%. The detection limits were 0.01-6.5 ng/ml (S/N=3). Our developed method is very useful for the simultaneous determination of five oxicam (non-steroidal anti-inflammatory) drugs in human plasma by LC/MS/MS.
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http://dx.doi.org/10.1016/j.forsciint.2012.11.016DOI Listing
April 2013

Simultaneous determination of 5 psychotropic drugs of various types in an autopsy case of acute multiple drug poisoning.

Forensic Sci Int 2013 Apr 23;227(1-3):90-4. Epub 2012 Dec 23.

Department of Legal Medicine, Kitasato University School of Medicine, 1-15-1 Kitasato, Sagamihara, Kanagawa, Japan.

We attempted the simultaneous determination of 5 drugs, mirtazapine, sertraline, chlorpromazine, amoxapine and zolpidem, detected in a gas chromatography-mass spectrometry screening test in an autopsy case. The solid-phase extraction of the analytes from biological samples was achieved using Oasis(®)HLB cartridges (Waters, Milford, MA, USA). Gas chromatography was performed on a HP-5MS fused silica capillary column (30 m × 0.25 mm i.d., 0.25 μm film thickness, Agilent Technologies). The mass spectrometer was operated with an electron energy of 70 eV in electron impact mode. The qualitative and quantitative analyses were performed in full-scan mode and the selected ion monitoring mode, respectively. The total ion chromatogram showed good separation of these drugs. Linear graphs were obtained with good correlation coefficients for these drugs from 0.001 to 2.0 μg/mL (r(2)=0.9909-0.9986) using imipramine-d6 as an internal standard. The recoveries of these drugs were found to be 62.8-88.0% in spiked whole blood. Mirtazapine, sertraline, chlorpromazine, amoxapine and zolpidem were found in post-mortem samples of the deceased at concentrations of 2.67, 0.07, 0.25, 0.32 and 0.68 μg/mL, respectively. The concentration of mirtazapine was within the lethal level and those of amoxapine and zolpidem were within the toxic level. We diagnosed that the cause of death was acute multiple drug poisoning. The simple and practical procedure used in this study is useful for the simultaneous determination of psychotropic drugs of various types in post-mortem biological samples.
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http://dx.doi.org/10.1016/j.forsciint.2012.11.015DOI Listing
April 2013

ABO genotyping by TaqMan assay and allele frequencies in a Japanese population.

Leg Med (Tokyo) 2013 Mar 12;15(2):57-60. Epub 2012 Oct 12.

Department of Legal Medicine, Kitasato University School of Medicine, Kanagawa 252-0374, Japan.

ABO genotyping have become common tools for forensic casework. We developed a new rapid ABO genotyping method using a fast real-time PCR system with the TaqMan® Sample-to-SNP™ Kit. Eight single nucleotide polymorphism (SNP) sites in the ABO gene (nt 261, 297, 467, 657, 703, 829, 930 and 1061) were selected to determine the ABO genotypes. ABO genotypes were easily determined by examining allelic discrimination patterns. This method enabled analyses to be completed in about 1h per plate with no postmortem change influences. The detection limit in each SNP site was examined as 100pg per reaction. ABO genotyping from 1000 Japanese individuals was also examined to determine the distribution of ABO genotypes and allele frequencies. Thus, 31 genotypes were clearly identified, and these were controlled by four common and seven rare alleles. The power of discrimination, heterozygosity and polymorphism information contents were 0.913, 0.775 and 0.812, respectively. Therefore, selecting these eight SNP sites could be useful for high specific ABO genotyping. This rapid, sensitive and accurate genotyping method is useful for forensic casework.
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http://dx.doi.org/10.1016/j.legalmed.2012.08.009DOI Listing
March 2013

Tetanus following replantation of an amputated finger: a case report.

J Med Case Rep 2012 Oct 8;6:343. Epub 2012 Oct 8.

Division of Plastic and Reconstructive Surgery, National Nagasaki Medical Center, 1001-1 Kubara 2, Ohmura City, 856-8562, Japan.

Unlabelled:

Introduction: Tetanus is an infectious disease caused by tetanus toxin produced by Clostridium tetani and induces severe neurological manifestations. We treated a patient who developed tetanus during hospitalization for replantation of an amputated finger. To the best of our knowledge, this is the first published case report of such an entity.

Case Presentation: A 49-year-old Japanese man had an amputation of his right middle finger at the distal interphalangeal joint region in an accident at work. His middle finger was successfully replanted, but his fingertip was partially necrotized because of crushing and so additional reconstruction with a reverse digital arterial flap was performed 15 days after the injury. Tetanus developed 21 days after replantation of the middle finger, but symptoms remitted via rapid diagnosis and treatment.

Conclusions: In replantation after finger trauma with exposure of nerve and blood vessel bundles, concern over injuring nerves and blood vessels may prevent irrigation and debridement from being performed sufficiently; these treatments may have been insufficiently performed in this patient. It is likely that the replanted middle finger partially adhered, and Clostridium tetani colonized the partially necrotized region. Even when there is only limited soil contamination, administration of tetanus toxoid and anti-tetanus immunoglobulin is necessary when the fingers are injured outdoors and the finger nerves and blood vessels are exposed. The drugs should be administered just after replantation if the finger has been amputated. However, if clinicians pay attention to the possibility of tetanus development, treatment can be rapidly initiated.
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http://dx.doi.org/10.1186/1752-1947-6-343DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3492091PMC
October 2012

Reconstruction of total nasal defect including skin, bone, and lining, using a single free radial forearm osteocutaneous perforator flap.

Plast Reconstr Surg 2012 May;129(5):854e-857e

Department of Plastic and Reconstructive Surgery, Nagasaki University, and, Department of Plastic and Reconstructive Surgery, Clinical Research Center, National Hospital Organization Nagasaki Medical Center, Nagasaki, Japan (Fujioka) Department of Plastic and Reconstructive Surgery, National Hospital Organization Nagasaki Medical Center, Nagasaki, Japan (Hayashida, Murakami, Koga).

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http://dx.doi.org/10.1097/PRS.0b013e31824a9e7fDOI Listing
May 2012

Evaluation of superficial and deep self-inflicted wrist and forearm lacerations.

J Hand Surg Am 2012 May 2;37(5):1054-8. Epub 2012 Mar 2.

Department of Plastic and Reconstructive Surgery, National Hospital Organization Nagasaki Medical Center, Nagasaki, Japan.

Purpose: Self-inflicted wrist or forearm laceration is a specific type of injury presenting to emergency departments. Many investigators have described wrist-cutting from a psychiatric viewpoint. We hypothesized that the character of patients with deep wounds is different from those with superficial wounds. We investigated patients who cut their wrist or forearms as an act of self-mutilation from the viewpoint of wound severity.

Methods: We reviewed 31 patients with self-inflected wrist injuries who were treated in our medical center from 2004 through 2009. We divided them into 2 groups: deep (15 patients) and superficial (16 patients). We investigated differences in age and gender, sites of self-cutting, frequency of self-injury attempts, object used for wrist cutting, group psychiatric parameters, required wound treatments, and psychiatric history and follow-up.

Results: Younger patients were more likely to have injured themselves severely compared with older patients. Differences in clinical findings between deep and superficial injury groups included the following: (1) all male patients had deep injuries; (2) patients with superficial wounds were more likely to have cut themselves previously; (3) patients in the deep injury group tended to injure themselves at multiple sites; (4) patients in the deep injury group tended to perform self-cutting with any sharp-edged object at hand; (5) 50% of our patients had received no psychiatric care before being seen by us for their injury; and (6) one-third discontinued the psychiatric treatment prematurely.

Conclusions: There are differences between patients who perform self-inflicted deep versus superficial wrist cutting. We also found that the ages and psychiatric diagnoses of our patients differed from previous reports. This is likely because the available literature includes only patients who received psychiatric care. We found that 50% of our patients had received no psychiatric care, which highlights the importance of hand surgeons treating these patients to initiate psychiatric consultation.
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http://dx.doi.org/10.1016/j.jhsa.2011.12.040DOI Listing
May 2012

Analysis of the sarcomere protein gene mutation on cardiomyopathy - Mutations in the cardiac troponin I gene.

Leg Med (Tokyo) 2010 Nov;12(6):280-3

Department of Legal Medicine, Kitasato University School of Medicine, 1-15-1 kitasato minami-ku, Sagamihara, Kanagawa 252-0374, Japan.

Developments in the molecular genetic studies of cardiomyopathy (CM) have led to discovery of a large number of mutations in the genes encoding the sarcomeric proteins. In this study, comprehensive screening of TNNI3 was performed in 36 consented autopsy cases diagnosed as CM, in order to evaluate the prevalence of gene mutations in sudden death caused by CM. In DCM cases, a new missense mutation Pro16Thr was detected. A single nucleotide polymorphism at -8 position of intron 3 (IVS 3 -8 T>A) was identified, which had a significant difference in allele frequency between DCM and control cases. From these results, it was indicated that this study contribute to genetic based diagnosis, risk stratification and prevention of sudden death caused by CM.
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http://dx.doi.org/10.1016/j.legalmed.2010.07.002DOI Listing
November 2010

A case of V-A shunt catheters migration into the pulmonary artery.

Leg Med (Tokyo) 2009 Jan 12;11(1):25-9. Epub 2008 Sep 12.

Department of Legal Medicine, Kitasato University School of Medicine, Sagamihara, Kanagawa, Japan.

A man in his sixties, who developed CPA at home, was transferred to the emergency center. Since CT images revealed a tube-shaped foreign body in the pulmonary artery, pulmonary embolism was initially suspected; however, this did not lead to a definite diagnosis. Autopsy revealed that the foreign body in the cadaver was a fragment of a V-A shunt catheter implanted about 30 years previously for the treatment of hydrocephalus. Although fibrous adhesion of a part of the catheter to the pulmonary artery wall was seen, suggesting that a fracture of the catheter had occurred a long time before, it was not known when the fracture had occurred. Since no pulmonary arterial obstruction secondary to the catheter or new thrombi, which had been initially suspected, were observed, the cause of death was determined to be ischemic cardiac failure. A fracture of a shunt catheter may be typically associated with some clinical manifestations, which are often found and treated. In this case, however, no symptoms appeared and the fracture of the shunt catheter remained untreated for a long time. This case was therefore considered to be extremely rare, and is an example of how a serious iatrogenic disease could occur.
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http://dx.doi.org/10.1016/j.legalmed.2008.06.005DOI Listing
January 2009

Effect of carotenoids and ascorbic acid of Japanese persimmons on cellular lipid peroxidation in HepG2 cells.

Biofactors 2004 ;21(1-4):241-5

Faculty of Education, Wakayama University, Wakayama, Japan.

In this experiment, we examine the functional property of carotenoids; beta-cryptoxanthin (Cry), zeaxanthin (Zea), beta-carotene (Car)) and ascorbic acid (AsA). The accumulation amounts of Cry, Zea and Car in HepG2 cells cultured in the high concentration medium were larger than that in a low concentration. Further those accumulation amounts in long incubation time within 24 hours were greater than that in a shorter time. When the added carotenoid concentration, with or without hydrogen peroxide, increased from 0 to 5 microM in the culture medium, the thiobarbituric acid reaction substance (TBARS) values in the HepG2 cells decreased significantly (p < 0.05). The decrease of TBARS values shows the antioxidative property of the carotenoids. When AsA and Tocopherol(Toc) were added to the medium from 0 to 20 microM, the TBARS values, with or without hydrogen peroxide, decreased significantly with increasing concentrations of AsA and Toc respectively (p < 0.05). The decreased amount of TBARS in 5 microM Cry compared with control(0 microM) was the largest among 6 antioxidants (Cry, Car, Zea, Retinol(Ret), AsA, Toc) used in this experiment.
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http://dx.doi.org/10.1002/biof.552210146DOI Listing
April 2005

Pseudodeflectusin, a novel isochroman derivative from Aspergillus pseudodeflectus a parasite of the sea weed, Sargassum fusiform, as a selective human cancer cytotoxin.

Bioorg Med Chem Lett 2004 Jul;14(13):3539-43

Department of Applied Biological Science, Tokyo University of Science, Noda, Chiba 278-8510, Japan.

A new isochroman derivative named pseudodeflectusin was isolated from a culture broth of Aspergillus pseudodeflectus. The structure was determined by spectroscopic means as 9-hydroxy-7-methyl-2-(methylethylidine)-furano[3,2-H]isochroman-3-one. This compound exhibited cytotoxicity for several human cancer cell lines from the stomach (NUGC-3), cervix (HeLa-S3), and peripheral blood (HL-60), but did not affect those from the lung (A549) or colon (DLD-1). The LD50 value of this compound for HL-60 cells was 39 microM.
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http://dx.doi.org/10.1016/j.bmcl.2004.04.050DOI Listing
July 2004

A novel DNA topoisomerase inhibitor: dehydroebriconic acid, one of the lanostane-type triterpene acids from Poria cocos.

Cancer Sci 2004 Apr;95(4):354-60

Laboratory of Food and Nutritional Sciences, Department of Nutritional Science and High Technology Research Center, Kobe-Gakuin University, Kobe, Hyogo 651-2180, Japan.

Traditional Chinese medicinal plants are a treasure house for screening novel inhibitors of DNA polymerases and DNA topoisomerases from mammals; in the present study, nine lanostane-type triterpene acids were found in sclerotium of Poria cocos. Among the nine compounds, only dehydroebriconic acid could potently inhibit DNA topoisomerase II (topo II) activity (IC(50) = 4.6 microM), while the compound moderately inhibited the activities of DNA polymerases alpha, beta, gamma, delta, epsilon, eta, iota, kappa and lambda only from mammals, to similar extents. Another compound, dehydrotrametenonic acid, also showed moderate inhibitory effects against topo II (IC(50) = 37.5 microM) and weak effects against all the polymerases tested. Both compounds showed no inhibitory effect against topo I, higher plant (cauliflower) DNA polymerase I (alpha-like polymerase) or II (beta-like polymerase), calf thymus terminal deoxynucleotidyl transferase, human immunodeficiency virus type-1 reverse transcriptase, prokaryotic DNA polymerases such as the Klenow fragment of E. coli DNA polymerase I, Taq DNA polymerase and T4 DNA polymerase, or DNA metabolic enzymes such as T 7 RNA polymerase, T4 polynucleotide kinase and bovine deoxyribonuclease I. These findings suggest that dehydroebriconic acid and dehydrotrametenonic acid should be designated as topo II-preferential inhibitors, although they also moderately inhibited all the mammalian DNA polymerases tested. Both dehydrotrametenonic acid and dehydroebriconic acid could prevent the growth of human gastric cancer cells, and their LD(50) values were 63.6 and 38.4 microM, respectively. The cells were halted at the G1 phase in the cell cycle. The relation between the structure of triterpene acids and their inhibitory activities is discussed.
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http://dx.doi.org/10.1111/j.1349-7006.2004.tb03215.xDOI Listing
April 2004

Mechanism of cell cycle arrest by sulfoquinovosyl monoacylglycerol with a C18-saturated fatty acid (C18-SQMG).

Biochem Pharmacol 2004 Apr;67(7):1373-80

Laboratory of Food and Nutritional Science, Department of Nutritional Science, Kobe-Gakuin University, Nishi-ku, Kobe, Hyogo 651-2180, Japan.

We have screened the inhibitors of mammalian DNA polymerases from natural products, and in the process found that either sulfoglycolipids or sulfoquinovosyl monoacylglycerol with a C18-saturated fatty acid (C18-SQMG), potently and selectively inhibited the activity of mammalian DNA polymerase (pol) and moderately the pol alpha. C18-SQMG was a cancer cell growth suppressor and a promissive anti-tumor agent. The purpose of this study was to elucidate the cell growth inhibition mechanism of C18-SQMG using HeLa cells. Analyses of the cell cycle and cyclin expression suggested that C18-SQMG arrested the cell cycle at intra-S phase, and the inhibition manner of DNA replication by C18-SQMG was similar to that by hydroxyurea. However, the DNA replication block by C18-SQMG did not induce degradation of Cdc25A protein, which was required for the replication block by hydroxyurea. C18-SQMG somewhat delayed mitosis because it induced phosphorylation of protein kinases, such as checkpoint kinases 1 and 2. These results suggest that C18-SQMG at first blocked DNA replication at the S phase by inhibiting replicative DNA polymerases, such as alpha, and then as the result of the inhibition, the other checkpoint signals associated with the pol might have responded.
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http://dx.doi.org/10.1016/j.bcp.2003.12.004DOI Listing
April 2004

Inhibitory action of emulsified sulfoquinovosyl acylglycerol on mammalian DNA polymerases.

Lipids 2003 Oct;38(10):1065-74

Laboratory of Food and Nutritional Sciences, Kobe-Gakuin University, Nishi-ku, Kobe, Hyogo 651-2180, Japan.

We reported previously that sulfoquinovosyl diacylglycerol and sulfoquinovosyl monoacylglycerol (SQDG/SQMG) are potent inhibitors of mammalian DNA polymerases and DNA topoisomerase II, and can be potent immunosuppressive agents and anticancer chemotherapy agents [Matsumoto, Y., Sahara, H., Fujita T., Shimozawa, K., Takenouchi, M., Torigoe, T., Hanashima, S., Yamazaki, T., Takahashi, S., Sugawara, F., et al., An Immunosuppressive Effect by Synthetic Sulfonolipids Deduced from Sulfonoquinovosyl Diacylglycerols of Sea Urchin, Transplantation 74, 261-267 (2002); Sahara, H., Hanashima, S., Yamazaki, T., Takahashi, S., Sugawara, F., Ohtani, S., Ishikawa, M., Mizushina, Y., Ohta, K., Shimozawa, K., et al., Anti-tumor Effect of Chemically Synthesized Sulfolipids Based on Sea Urchin's Natural Sulfonoquinovosylmonoacylglycerols, Jpn. J. Cancer Res. 93, 85-92 (2002)]. In those experiments, the in vivo effectiveness greatly depended on the degree of water solubility of SQDG/SQMG. In the present work, we studied the emulsification of SQDG/SQMG in terms of their use in in vivo experiments. Lipid emulsions containing SQDG/SQMG (oil-in-water emulsions) in which the particle size was smaller than 100 nm were designed and synthesized, and then the biochemical modes of emulsified SQDG/SQMG were studied in comparison with those of SQDG/SQMG solubilized by DMSO. Emulsified SQDG/SQMG are also selective mammalian DNA polymerase inhibitors and potent antineoplastic agents but do not inhibit the DNA topoisomerase II activity. The growth inhibition effect of emulsified SQMG to NUGC-3 cancer cells was twofold stronger than DMSO-soluble SQMG (69 and 151 microM, respectively). From these results, the properties of lipid emulsions containing SQDG/SQMG and their possible use in in vivo experiments including clinical use are discussed.
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http://dx.doi.org/10.1007/s11745-006-1162-1DOI Listing
October 2003

Pyridoxal 5'-phosphate is a selective inhibitor in vivo of DNA polymerase alpha and epsilon.

Biochem Biophys Res Commun 2003 Dec;312(4):1025-32

Department of Nutritional Science, Kobe-Gakuin University, Nishi-ku, Kobe, 651-2180, Hyogo, Japan.

Vitamin B(6) compounds such as pyridoxal 5(')-phosphate (PLP), pyridoxal (PL), pyridoxine (PN), and pyridoxamine (PM), which reportedly have anti-angiogenic and anti-cancer effects, were thought to be inhibitors of some types of eukaryotic DNA polymerases. PL moderately inhibited only the activities of calf DNA polymerase alpha (pol alpha), while PN and PM had no inhibitory effects on any of the polymerases tested. On the other hand, PLP, a phosphated form of PL, was potentially a strong inhibitor of pol alpha and epsilon from phylogenetic-wide organisms including mammals, fish, insects, plants, and protists. PLP did not suppress the activities of prokaryotic DNA polymerases such as Escherichia coli DNA polymerase I and Taq DNA polymerase, or DNA-metabolic enzymes such as deoxyribonuclease I. For pol alpha and epsilon, PLP acted non-competitively with the DNA template-primer and competitively with the nucleotide substrate. Since PL was converted to PLP in vivo after being incorporated into human cancer cells, the anti-angiogenic and anti-cancer effects caused by PL must have been caused by the inhibition of pol alpha and epsilon activities after conversion to PLP.
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http://dx.doi.org/10.1016/j.bbrc.2003.11.027DOI Listing
December 2003

Some anti-chronic inflammatory compounds are DNA polymerase lambda-specific inhibitors.

Biochem Pharmacol 2003 Nov;66(10):1935-44

Department of Nutritional Science, Kobe-Gakuin University, Nishi-ku, Kobe, 651-2180 Hyogo, Japan.

We previously reported that a phenolic compound, petasiphenol, was a selective inhibitor of DNA polymerase lambda (pol lambda) in vitro. We found here that another phenolic compound, curcumin (diferuloylmethane), which is known as an anti-chronic inflammatory agent and is structurally quite similar to petasiphenol, was also a potent pol lambda inhibitor. The IC(50) values of petasiphenol and curcumin were 7.8 and 7.0 microM, respectively. Curcumin, as well as petasiphenol, did not influence the activities of replicative DNA polymerases, such as alpha, gamma, delta, and epsilon, but also showed no effect even on the pol beta activity belonging to the X family. Curcumin could prevent the growth of human NUGC-3 cancer cells with LD(50) values of 13 microM, and halted them at the G2/M phase in the cell cycle, whereas petasiphenol suppressed the cell growth at 66 microM and arrested the cells at the G1 phase. These data showed that curcumin and petasiphenol were slightly different functionally. We also previously reported that novel anti-inflammatory terpeno benzoic acids and triterpenoids were inhibitors of mammalian DNA polymerases. They could also efficiently inhibit the pol lambda activity, although they influenced the other polymerase species to the same extent, suggesting that there may be a physiological relationship between pol lambda inhibition and anti-12-O-tetradecanoylphorbol-13-acetate-induced inflammation. Expectedly, petasiphenol also showed an anti-12-O-tetradecanoylphorbol-13-acetate-induced inflammatory effect in mice. This finding may provide clues to investigating the molecular mechanism of inflammation.
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http://dx.doi.org/10.1016/s0006-2952(03)00551-3DOI Listing
November 2003

Analysis of cell cycle regulation by 1-mono-O-acyl-3-O-(alpha-D-sulfoquinovosyl)-glyceride (SQMG), an inhibitor of eukaryotic DNA polymerases.

Biochem Pharmacol 2003 Aug;66(4):541-50

Laboratory of Food and Nutritional Sciences, Department of Nutritional Science, Kobe-Gakuin University, Nishi-ku, Kobe, Hyogo 651-2180, Japan.

One of the sulfo-lipids, 1-mono-O-acyl-3-O-(alpha-D-sulfoquinovosyl)-glyceride (SQMG), potently and selectively inhibited the activity of mammalian DNA polymerases. SQMG was also a potent apoptosis inducer and the SQMG effect occurred through the induction of G1 arrest with a reduction in the proportion of cells in the S phase. SQMG clearly increased the levels of p53 and p21 proteins, but did not induce the expression of p27 and p16 proteins. SQMG markedly reduced the pRb protein level and inhibited pRb phosphorylation after 48hr. These results suggested that SQMG activates the G1 checkpoint as a result of the DNA polymerase inhibition, and then promotes a p53-dependent apoptotic response. Since aphidicolin, a well-known replicative DNA polymerase inhibitor, did not promote these protein expressions, the apoptosis-inducing pathway by SQMG differs from that by aphidicolin.
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http://dx.doi.org/10.1016/s0006-2952(03)00345-9DOI Listing
August 2003

Selective inhibition of mammalian DNA polymerase alpha by vitamin D2 and D3.

J Pharmacol Sci 2003 Jul;92(3):283-90

Laboratory of Food & Nutritional Sciences, Department of Nutritional Science, Kobe-Gakuin University, Hyogo, Japan.

As described previously (H. Togashi et al. Biochem Pharmacol. 1998;56:583-590), the irradiated products of provitamin D(2) (ergosterol) inhibit the activities of eukaryotic DNA polymerases. In this report, therefore, we investigated whether vitamin D and its related compounds inhibited the activities of DNA polymerases. As expected, vitamin D(2) and vitamin D(3) were found to be selective inhibitors of mammalian DNA polymerase alpha (pol alpha) with IC(50) values of 123 and 96 micro M, respectively. On the other hand, provitamin D(2), provitamin D(3), and the active form of vitamin D(3) such as 1alpha,25-dihydroxyvitamin D(3) could not influence any of the DNA polymerase activities. Interestingly, vitamin D(3)-3beta-sulfate was a much stronger pol alpha inhibitor with an IC(50) value of 7.1 micro M. Vitamin D(2), vitamin D(3), and vitamin D(3)-3beta-sulfate could prevent the growth of NUGC-3 human gastric cancer cells with LD(50) values of 133, 77, and 44 micro M, respectively, but provitamin D(2) and provitamin D(3) could not. The cells were halted at the G1 phase in the cell cycle by these compounds.
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http://dx.doi.org/10.1254/jphs.92.283DOI Listing
July 2003

Distinct enzymatic properties of recombinant mouse DNA methyltransferases Dnmt3a and Dnmt3b.

J Biochem 2003 Jun;133(6):737-44

Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871.

Recombinant mouse Dnmt3a and Dnmt3b were expressed in sf9 cells and purified to near homogeneity. The purified Dnmt3a and Dnmt3b gave specific activities of 1.8 +/- 0.3 and 1.3 +/- 0.1 mol/h/mol enzyme towards poly(dGdC)-poly(dGdC), respectively, which were the highest among those reported. Dnmt3a or Dnmt3b showed similar K(m) values towards poly(dIdC)-poly(dIdC) and poly(dGdC)-poly(dGdC). The K(m) values for S-adenosyl-L-methionine were not affected by the methyl-group acceptors, poly(dI-dC)-poly(dIdC) and poly(dG-dC)-poly(dGdC). The results indicate that the enzymes are de novo-type DNA methyltransferases. Dnmt3a and Dnmt3b activities were inhibited by Mn(2+) and Ni(2+) and showed broad pH optima around neutral pH. Both enzymes were susceptible to sodium ions, which inhibited their activity at around physiological ionic strength. However, Dnmt3a was fully active at physiological potassium concentration, but Dnmt3b was not. Using designed oligonucleotides for the analysis of cytosine methylation, we demonstrated that, in addition to CpG, Dnmt3a methylated CpA but not CpT and CpC, and that Dnmt3b methylated CpA and CpT but scarcely CpC. The relative activity of Dnmt3b towards nonCpG sequences was higher than that of Dnmt3a. These differences in enzymatic properties of Dnmt3a and Dnmt3b may contribute to the distinct functions of these enzymes in vivo.
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http://dx.doi.org/10.1093/jb/mvg095DOI Listing
June 2003

Molecular action mode of Hippospongic acid A, an inhibitor of gastrulation of starfish embryos.

J Biochem 2003 Apr;133(4):541-52

Department of Nutritional Science, Kobe-Gakuin University, Nishi-ku, Kobe, Hyogo 651-2180.

Hippospongic acid A (HA-A) is a novel natural triterpene metabolite that exhibits inhibitory activity against the gastrulation of starfish embryos isolated from a marine sponge, Hippospongia sp. We succeeded in chemically synthesizing the natural enantiomer and the racemate HA-A. In this study, we examined its action mode in vitro. HA-A was a rare compound that could selectively but uniformly inhibit the activities of all the vertebrate DNA polymerases tested such as alpha, beta, delta, epsilon, eta, kappa, and lambda, in the IC(50) range of 5.9-17.6 microM, and interestingly also those of human DNA topoisomerases I and II (IC(50) = 15-25 microM). HA-A exhibited no inhibitory effect on DNA polymerases from insects, plants and prokaryotes, or on many other DNA metabolic enzymes. HA-A was an inhibitor specific to DNA polymerases and DNA topoisomerases from vertebrates, but not selective as to a subclass species among the enzymes. Since DNA polymerase beta is the smallest, we used it to analyze the biochemical relationship with HA-A. Biochemical, BIAcore and computer modeling analyses demonstrated that HA-A bound selectively to the N-terminal 8 kDa DNA template-binding domain of DNA polymerase beta, and HA-A inhibited the ssDNA binding activity. HA-A could prevent the growth of NUGC-3 cancer cells at both the G1 and G2/M phases, and induce apoptosis in the cells. The LD(50) value was 9.5 microM, i.e. in the same range as for the enzyme inhibition. Therefore, we concluded that one molecular basis of the gastrulation of starfish embryos is a process that requires DNA polymerases and DNA topoisomerases, and subsequently the gastrulation was inhibited by HA-A. We also discussed the in vivo role of HA-A.
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http://dx.doi.org/10.1093/jb/mvg070DOI Listing
April 2003

Kohamaic acid A, a novel sesterterpenic acid, inhibits activities of DNA polymerases from deuterostomes.

Biochim Biophys Acta 2003 May;1648(1-2):55-61

Laboratory of Food and Nutritional Sciences, Department of Nutritional Science, Kobe-Gakuin University, Nishi, Hyogo 651-2180, Japan.

We previously found and isolated a novel natural product, designated kohamaic acid A (KA-A), which inhibited the first cleavage of fertilized sea urchin eggs. In this paper, we report that this compound could selectively inhibit the activities of DNA polymerases (pol. alpha, beta, gamma, delta and epsilon ) only from species in the deuterostome branch in the animal kingdom, like sea urchin, fish and mammals, but not from protostomes including insects (fruit fly, Drosophila melanogaster) and mollusks (octopus and oyster). Inhibition of deuterostome DNA polymerases was dose dependent. IC(50) values for DNA polymerases of mammals and fish occurred at approximately 5.8-14.9 microM and those of sea urchin at 6.1-30.3 microM. In the sea urchin DNA polymerases, the activities of the replicative DNA polymerases such as alpha, delta and epsilon were more strongly inhibited than that of the repair-related pol. beta. KA-A is an inhibitor of replicative DNA polymerases from the deuterostome species, and subsequently, the inhibition of the first cleavage of fertilized sea urchin eggs might occur as a result of the suppression of DNA replication.
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http://dx.doi.org/10.1016/s1570-9639(03)00108-0DOI Listing
May 2003

A novel DNA polymerase inhibitor and a potent apoptosis inducer: 2-mono-O-acyl-3-O-(alpha-D-sulfoquinovosyl)-glyceride with stearic acid.

Biochim Biophys Acta 2003 Jan;1645(1):72-80

Department of Nutritional Science, Laboratory of Food & Nutritional Sciences, Kobe-Gakuin University, Nishi-ku, Hyogo 651-2180, Kobe, Japan.

Sulfo-glycolipids in the class of sulfoquinovosyl diacylglycerol (SQDG) including the stereoisomers are potent inhibitors of DNA polymerase alpha and beta. However, since the alpha-configuration of SQDG with two stearic acids (alpha-SQDG-C(18)) can hardly penetrate cells, it has no cytotoxic effect. We tried and succeeded in making a permeable form, sulfoquinovosyl monoacylglycerol with a stearic acid (alpha-SQMG-C(18)) from alpha-SQDG-C(18) by hydrolysis with a pancreatic lipase. alpha-SQMG-C(18) inhibited DNA polymerase activity and was found to be a potent inhibitor of the growth of NUGC-3 cancer cells. alpha-SQMG-C(18) arrested the cell cycle at the G1 phase, and subsequently induced severe apoptosis. The arrest was correlated with an increased expression of p53 and cyclin E, indicating that alpha-SQMG-C(18) induced cell death through a p53-dependent apoptotic pathway.
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http://dx.doi.org/10.1016/s1570-9639(02)00521-6DOI Listing
January 2003

Effects of glycolipids from spinach on mammalian DNA polymerases.

Biochem Pharmacol 2003 Jan;65(2):259-67

Laboratory of Food and Nutritional Sciences, Department of Nutritional Science, Kobe-Gakuin University, Nishi-ku, Kobe, Hyogo 651-2180, Japan.

We purified the major glycolipids in the class of monogalactosyl diacylglycerol (MGDG), digalactosyl diacylglycerol (DGDG) and sulfoquinovosyl diacylglycerol (SQDG) from a green vegetable, spinach (Spinacia oleracea L.). MGDG was an inhibitor of the growth of NUGC-3 human gastric cancer cells, but DGDG and SQDG had no such cytotoxic effect. Therefore, we studied MGDG and its monoacyglycerol-form, monogalactosyl monoacylglycerol (MGMG), in detail. MGMG with one fatty acid molecule was obtained from MGDG with two fatty acid molecules by hydrolyzing with a pancreatic lipase. MGMG was also found to prevent the cancer cell growth. MGDG was a potent inhibitor of replicative DNA polymerases such as alpha, delta and epsilon. MGMG inhibited the activities of all mammalian DNA polymerases including repair-related DNA polymerase beta with IC(50) values of 8.5-36 microg/mL, and the inhibition by MGMG was stronger than that by MGDG. Both MGDG and MGMG could halt the cell cycle at the G1 phase, and subsequently induced severe apoptosis. The relationship between the DNA polymerase inhibition and the cell growth effect by these glycolipids is discussed.
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http://dx.doi.org/10.1016/s0006-2952(02)01483-1DOI Listing
January 2003