Toda la información sobre las últimas publicaciones científicas de la Clínica Universidad de Navarra. Transduction efficacy, antitumoral effect, and toxicity of adenovirus-mediated herpes simplex virus thymidine kinase/ ganciclovir therapy of hepatocellular carcinoma: the woodchuck animal model
All information about the latest scientific publications of the Clínica Universidad de Navarra. Transduction efficacy, antitumoral effect, and toxicity of adenovirus-mediated herpes simplex virus thymidine kinase/ ganciclovir therapy of hepatocellular carcinoma: the woodchuck animal model
Abstract We constructed a functional MoMuLV-based bicistronic retroviral vector encoding the herpes simplex virus type I thymidine kinase gene, which induces sensitivity to the prodrug ganciclovir (gcv), and the reporter beta-galactosidase gene (MFG-tk-IRES-lacZ). The U937 histiocytic cell line was transduced with this vector, and a clone (VB71) with high-level transgene expression was selected. Severe combined immunodeficient (SCID) mice were injected with VB71 cells to evaluate the role of long terminal repeat methylation in transgene silencing in vivo and to see whether 5-azacytidine (5 aza-C) demethylating agent prevented it. We found 5 aza-C maintained gene expression at high level in vitro. In vivo, time to tumor onset was significantly longer in SCID mice receiving the VB71 cells, 5 aza-C, and gcv compared with animals treated with either 5 aza-C or gcv alone. The number of injected tumor cells influences tumor onset time and the efficacy of 5 aza-C and gcv treatment. The standard ...
Summary Purified herpes simplex virus thymidine kinase has been used to immunize mice for the production of monoclonal antibodies to the enzyme. Monoclonal antibodies were successfully produced against both herpes simplex virus type 1 and type 2 enzymes. These antibodies should prove useful for detecting the enzyme under a variety of experimental conditions. We also demonstrate that the antibodies can provide an alternative method for obtaining large amounts of purified thymidine kinase.
TY - JOUR. T1 - Stem cells as vectors to deliver HSV/tk gene therapy for malignant gliomas. AU - Rath, Prakash. AU - Shi, Huidong. AU - Maruniak, Joel A.. AU - Litofsky, N. Scott. AU - Maria, Bernard L.. AU - Kirk, Mark D.. PY - 2009/10/12. Y1 - 2009/10/12. N2 - The prognosis of patients diagnosed with malignant gliomas including glioblastoma multiforme (GBM) is poor and there is an urgent need to develop and translate novel therapies into the clinic. Neural stem cells display remarkable tropism toward GBMs and thus may provide a platform to deliver oncolytic agents to improve survival. First we provide a brief review of clinical trials that have used intra-tumoral herpes simplex virus thymidine kinase (HSV/tk) gene therapy to treat brain tumors. Then, we review recent evidence that neural stem cells can be used to deliver HSV/tk to GBMs in animal models. While previous clinical trials used viruses or non-migratory vector-producing cells to deliver HSV/tk, the latter approaches were not ...
Summary The induction of thymidine kinase (TK) and DNA polymerase was inhibited by interferon (IFN) in mouse L-cells infected with herpes simplex virus type 1 (HSV-1). The inhibitory activity of IFN at this early stage of HSV-1 replication was followed by a reduced virus yield and was dependent on the multiplicity of infection. The expression of a cloned thymidine kinase (tk) gene of HSV-1, in biochemically transformed L-cells (LTK+), was not affected by IFN. These same LTK+ cells, however, developed an antiviral state since, upon HSV-1 infection, the induction of TK and DNA polymerase of the replicating virus was inhibited by IFN. Furthermore, IFN inhibited the transactivation of the HSV-1 tk gene in the biochemically transformed LTK+ cells, which followed infection by a virus mutant defective in the tk gene (HSV-1 TK-). This transactivation is dependent on expression of immediate-early HSV-1 α-genes. These results indicate that IFN inhibits HSV-1 replication at an early step prior to DNA synthesis.
Thymidine kinases form part of the salvage pathway for pyrimidine deoxyribonucleotide synthesis. TKs are expressed in a variety of organisms from human to bacteria as well as in a number of viruses. The reaction catalysed by TK involves the transfer of a γ-phosphoryl moiety from ATP to 2deoxy-thymidine (dThd) to produce thymidine 5-monophosphate (dTMP). Certain TKs, such as those from herpes simplex virus type 1 (HSV-1) and varicella zoster virus (VZV) have, in addition, thymidylate kinase activity allowing the conversion of dTMP to thymidine 5-diphosphate (dTDP). TKs can be classified into two types which differ in several respects [1]. Type 1 TKs are of higher molecular weight, typically around 40 kDa, and are active as homodimers. This subfamily contains the HSV1, HSV2 and VZV TKs, and also mitochondrial TK.. TKs of type 2 include those from poxviridae such as vaccinia virus (VV) and variola virus, [2], as well as from human [3] hTK, (human type II thymidine kinase 1) and mouse [4]. Type ...
Mammals have two isoenzymes, that are chemically very different, TK1 and TK2. The former was first found in fetal tissue, the second was found to be more abundant in adult tissue, and initially they were termed fetal and adult thymidine kinase. Soon it was shown that TK1 is present in the cytoplasm only in anticipation of cell division (cell cycle-dependent),[10][11] whereas TK2 is located in mitochondria and is cell cycle-independent.[12][13] The two isoenzymes have different reaction kinetics and are inhibited by different inhibitors. The viral thymidine kinases differ completely from the mammalian enzymes both structurally and biochemically and are inhibited by inhibitors that do not inhibit the mammalian enzymes.[14][15][16] The genes of the two human isoenzymes were localized in the mid-1970s.[17][18] The gene for TK1 was cloned and sequenced.[19] The corresponding protein has a molecular weight of about 25 kD. Normally, it occurs in tissue as a dimer with a molecular weight of around 50 ...
Aims: Thymidine kinase 1 (TK1) is an enzyme involved in DNA synthesis and an important proliferation marker. We explored the association of preoperative serum TK1 activity with clinicopathological parameters and prognosis in terms of recurrence-free survival (RFS) in breast cancer (BC) patients. Patients and methods: TK1 activity in serum of 120 healthy women and 161 BC patients was measured by quantitative ELISA. Results: Serum TK1 activity in BC patients was significantly higher than in healthy women (P , 0.0001). In BC patients elevated TK1 activity was significantly associated with advanced T stage (P = 0.015), higher grade (P = 0.013), presence of tumor necrosis (P = 0.006), vascular invasion (P = 0.002), and lack of estrogen receptor (ER) and progesterone receptor (PR) expression (P = 0.0004 and P = 0.003). Higher TK1 activity was found in patients with BRCA1/2 mutations compared to those without the mutation (P = 0.004). Multivariate Cox proportional hazards analyses demonstrated that ...
TY - JOUR. T1 - Differences in thermostability of thymidine kinase isoenzymes in normal ovary and ovarian carcinoma. AU - Demeter, A.. AU - Abonyi, M.. AU - Look, K. Y.. AU - Keszler, G.. AU - Staub, M.. AU - Weber, G.. PY - 2001/4/17. Y1 - 2001/4/17. N2 - Thymidine kinase I (TK 1 EC. 2.7.1.21) the most specific and cell-cycle regulated salvage enzyme for pyrimidine nucleoside supply of DNA synthesis is a promising target to rationally designed chemo- and other therapies. The present study was undertaken to compare the heat stability of TK isoenzymes of both normal ovarian and epithelial ovarian cancer cells. Tissue extracts of epithelial ovarian carcinomas (N = 7) and normal ovaries (N=9) were analyzed for thymidine kinase activity using the polyethyleneimine-cellulose disc radioassay. The TK activity in extracts of ovarian carcinomas was 12-fold higher than in extracts of normal ovaries. The TK activity of ovarian carcinomas decreased significantly even after 30 minutes incubation at 37°C ...
DI-fusion, le Dépôt institutionnel numérique de lULB, est loutil de référencementde la production scientifique de lULB.Linterface de recherche DI-fusion permet de consulter les publications des chercheurs de lULB et les thèses qui y ont été défendues.
9-(4-(18)F-Fluoro-3-[hydroxymethyl]butyl)guanine ((18)F-FHBG) is a sensitive and specific PET reporter probe for imaging the PET reporter genes, herpes simplex 1 thymidine kinase (HSV1-tk) and its mutant HSV1-sr39tk. (18)F-FHBG has suitable pharmacok
TY - JOUR. T1 - Glucocorticoid regulation of the genes encoding thymidine kinase, thymidylate synthase, and ornithine decarboxylase in P1798 cells. AU - Barbour, Karen W.. AU - Berger, Sondra H.. AU - Berger, Franklin G.. AU - Thompson, E. Aubrey. PY - 1988/1. Y1 - 1988/1. N2 - The expression of a number of genes was measured in P1798 cells treated for various periods of time with 0.1 mm dexamethasone. Thymidine kinase (TK) activity decreased under these conditions with 50% inhibition achieved within approximately 8 h. Decreased TK activity was associated with reduced abundance of TK mRNA. Analysis of nuclear tran-scription indicated that this was attributable to a decrease in the number of RNA polymerase II molecules engaged in transcription of the TK gene. With respect to TK, there was an overall correlation between enzyme activity, mRNA, and nuclear transcription. The data are consistent with the hypothesis that glucocorticoid inhibition of expression of TK is primarily due to inhibition of ...
TY - JOUR. T1 - Gene therapy for transplant arteriosclerosis. AU - Rekhter, M. D.. AU - Shah, N.. AU - Simari, R. D.. AU - Work, C.. AU - Kim, J. S.. AU - Nabel, G. J.. AU - Nabel, E. G.. AU - Gordon, D.. PY - 1997/12/1. Y1 - 1997/12/1. N2 - Transplant arteriosclerosis is a thickening of the intima that develops in arteries of transplanted organs. Smooth muscle cell (SMC) replication plays a central role in its pathogenesis. One strategy to selectively eliminate dividing cells is to express a herpesvirus thymidine kinase (tk) gene which phosphorylates the nucleoside analog, ganciclovir, into a toxic form which leads to cell killing. To evaluate a feasibility of transgene expression in arterial grafts, we performed ex vivo adenovirus (ADV)-mediated transfer of human placental alkaline phosphatase (hpAP) gene into aortas of donor rabbits, and then transplanted them into carotid artery of recipient rabbits. 2 days post-transplant luminal endothelial cells, adventitial cells, and medial SMCs ...
Local inflammatory response and vector spread after direct intraprostatic injection of a recombinant adenovirus containing the herpes simplex virus thymidine kinase gene and ganciclovir therapy in mice Academic Article ...
DNA containing the herpes simplex virus thymidine kinase (HSVtk) gene was used to transform wild-type tk+ mouse L cells to a tk++ status in vitro using methotrexate as a selective agent. HSVtk DNA was also used to transform mouse bone marrow cells in vitro. Transformed marrow cells injected into irradiated and methotrexate-treated recipient mice gave rise to proliferating cells which in some cases dominated the marrow population and which contained HSVtk gene sequences. ...
Cadee JA, van Luyn MJ, Brouwer LA, Plantinga JA, van Wachem PB, De Groot CJ, den OW, and Hennink WE. (2000). In vivo biocompatibility of dextran-based hydrogels. J Biomed Mater Res 50: 397-404. PubMed. de Vries EF, van WA, Harmsen MC, Mulder NH, Vaalburg W, and Hospers GA. (2000). [(11)C]FMAU and [(18)F]FHPG as PET tracers for herpes simplex virus thymidine kinase enzyme activity and human cytomegalovirus infections. Nucl Med Biol 27: 113-9. PubMed. Dijk F, Westerhof M, Busscher HJ, van Luyn MJ, and van Der Mei HC. (2000). In vitro formation of oropharyngeal biofilms on silicone rubber treated with a palladium/tin salt mixture. J Biomed Mater Res 51: 408-12. PubMed. Kas-Deelen AM, de Maar EF, Harmsen MC, Driessen C, van Son WJ, and The TH. (2000). Uninfected and cytomegalic endothelial cells in blood during cytomegalovirus infection: effect of acute rejection. J Infect Dis 181: 721-4. PubMed. Kas-Deelen AM, Harmsen MC, de Maar EF, Oost-Kort WW, Tervaert JW, Van Der Meer J, van Son WJ, and The ...
EBV Thymidine Kinase, 0.1 mg. Thymidine kinase (TK) belongs to a group of enzymes such as dihydrofolate reductase, thymidylate synthase, and DNA polymerase that are involved in DNA synthesis and precursor production.
Valaciclovir belongs to a family of molecules. Valaciclovir is a prodrug, an esterified version of aciclovir that has greater oral bioavailability (about 55%) than aciclovir (10-20%). It is converted by esterases to the active drug aciclovir, as well as the amino acid valine, via hepatic first-pass metabolism. Aciclovir is selectively converted into a monophosphate form by viral thymidine kinase, which is far more effective (3000 times) in phosphorylation of aciclovir than cellular thymidine kinase. Subsequently, the monophosphate form is further phosphorylated into the active triphosphate form, aciclo-GTP, by cellular kinases. Aciclo-GTP is a very potent inhibitor of viral DNA polymerase; it has approximately 100 times higher affinity to viral than cellular polymerase.[citation needed] Its monophosphate form also incorporates into the viral DNA, resulting in chain termination. It has also been shown that the viral enzymes cannot remove aciclo-GMP from the chain, which results in inhibition of ...
Although acyclovir (ACV) is an effective antiviral agent, herpes simplex virus (HSV) that is resistant to treatment can be a serious clinical problem in immunocompromised patients. It has been estimated that ACV-resistant (ACVr) HSV appears in as many as 5% of this population upon treatment (4, 10). Clinically significant drug resistance implies evasion of drug action but retention of pathogenesis. The vast majority of mutations associated with clinical drug resistance occur in the viral thymidine kinase (TK) gene (tk), which encodes the enzyme that selectively activates ACV. The phenotypes of clinical isolates with mutant TKs typically fall into one of three categories: altered substrate specificity, low TK activity (TKL), or lack of measurable TK activity (TK−) (11, 12, 26). That viruses lacking TK activity are pathogenic has been puzzling. Although TK is not required for growth in cell culture, it is essential for pathogenesis in mouse models of HSV infection, in particular, for ...
Most individuals have no or only minimal signs or symptoms from HSV-1 or HSV-2 infection. ELLANSÉ™ with its four distinctive versions, lasting from 12 months up to 4 years, allow you to choose how long you would like the results to last, eliminating the need for frequent repeat treatments. Herpes infections are common among women of reproductive age (i.e., aged 15-44 years). Patients older than 65 years are usually considered ineligible for ASCT. In U. The sympathetic fibres parallel the parasympathetic fibres as they supply the same areas. Herpes simplex virus thymidine kinase (HSV-TK) is widely used in gene therapy.. TheBody. First, a lot of time and energy worrying that your partner will get herpes. The culprit responsible for cold sores is the herpes simplex virus. IgM antibodies are produced immediately after infection. What electrodesiccation uses in destroying chlamydia is an electric current. Make research projects and school reports about Canker Sores easy with credible articles ...
It is called suicide gene therapy. And this is the concept: find a gene coding for an enzyme that human cells dont have. Insert that gene only into cancer cells--not normal cells. This enzyme works by taking a harmless "pro-drug" and converting it into a toxic compound. Early attempts at this therapy used the enzyme thymidine kinase from the herpes simplex virus. This enzyme converts the drug ganciclovir into a compound that stops DNA synthesis. If a cancer cell cant make new DNA, it will eventually die ...
Viral infection of mammalian cells entails the regulated induction of viral gene expression. The induction of many viral genes, including the herpes simplex virus gene encoding thymidine kinase (tk), depends on viral regulatory proteins that act in trans. Because recognition of the tk promoter by cellular transcription factors is well understood, its trans induction by viral regulatory proteins may serve as a useful model for the regulation of eukaryotic gene expression. A comprehensive set of mutations was therefore introduced into the chromosome of herpes simplex virus at the tk promoter to directly analyze the effects of promoter mutations on tk transcription. The promoter domains required for efficient tk expression under conditions of trans induction corresponded to those important for recognition by cellular transcription factors. Thus, trans induction of tk expression may be catalyzed initially by the interaction of viral regulatory proteins with cellular transcription factors. ...
Figure and Images: Expression of functional MMP-9 by GLV-1h255-infected tumor cells. (A) Expression cassettes of GLV-1h68 and GLV-1h255. In GLV-1h255 the insert in the Tk locus was replaced by the human mmp-9 gene under control of the PSE promoter. PSEL, synthetic early/late promoter; PSE, synthetic early promoter; P7.5, VACV p7.5 K early/late promoter; P11, VACV p11 late promoter; Tk, thymidine kinase locus, Ha, hemagglutinin locus. (B) Expression of virus-encoded MMP-9 (92 kDa) in GLV-1h255 infected PC-3 cells and supernatants in vitro, β-actin (42 kDa) was used as a loading control. (C) Activity of the MMP-9 protein was tested by gelatin zymography. Lysates and supernatants of infected A549 cells were isolated and separated by non-reducing SDS-PAGE. In zymography, cleavage of the substrate by MMP-9 resulted in a clear band. ...
BioAssay record AID 221799 submitted by ChEMBL: Compound was tested for antitumor activity against thymidine kinase deficient human B-lymphoblast Raji/0 cells.
Acquistare indescribably forzest 20 buy online gli integratori che elencheremo in questo report è al 100% legale per la Legge Italiana. In another embodiment, a fusion protein comprised of a modified form of the tet repressor (TetR) and a transactivation domain or a domain (eg, a dimerization domain) which recruits a transcriptional activator (eg, an endogenous transcriptional activator) to interact with the fusion protein by a protein-protein interaction (eg, a non-covalent interaction) is introduced into a cell. Coli is also found in this environment ( 60, 61). Viral resistance to acyclovir may occur due to loss of thymidine kinase activity, levitra malaysia alterations in thymidine kinase substrate specificity, or decreased DNA-polymerase sensitivity. I have heard excellent things about blogenginenet! Im not very internet smart so Im not 100% certain? «Lady Gaga dedicates her new Little Monsters tattoo to her fans». The decision making it is only has not a new buy retin a cream ...
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1E2P: The Effect of Substrate Binding on the Conformation and Structural Stability of Herpes Simplex Virus Type 1 Thymidine Kinase
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FLT was developed to provide a measure of tumor proliferation because it is taken up by cells and retained after phosphorylation by thymidine kinase 1. Thymidine kinase 1 increases as cells enter the DNA synthetic phase, and previous work has shown that FLT retention generally correlates with other measures of tumor proliferation. The most likely use of FLT will be in monitoring treatment response, but such studies need to consider the different methods for measuring FLT retention in tumors and their reproducibility.. The most common method of measuring tracer retention has been through the use of the semiquantitative approach of the SUV. The SUV uses the measurement of activity within a ROI along with the injected dose and the weight of the patient to calculate the desired variable. To take into account differences in the proportion of fat in patients, some investigators have adjusted the SUV by using lean body mass, ideal body weight, or even body surface area. When studying the ...
srp:SSUST1_0992 K00857 thymidine kinase [EC:2.7.1.21] , (GenBank) tdk; thymidine kinase (A) MAQLYYKYGTMNSGKTIEILKVAHNYEEQGKPVVIMTSALDTRDAFGVVSSRIGMRRDAV AIDDDMDIFGFIEKMEPRPYCVLIDEAQFLRRHHVYALARVVDELDVPVMAFGLKNDFRN ELFEGSQHLLLLADKLDEIKTICQYCSKKATMVLRTQDGKPTYEGEQIQIGGNETYIPVC RKHYFSPEIKELP ...
swd:Swoo_3233 K00857 thymidine kinase [EC:2.7.1.21] , (GenBank) Thymidine kinase (A) MAQLYFYYSAMNAGKSTSLLQSSYNYRERGMNTLVMTASIDDRYGVGKVASRIGIETDAQ VFGSDDNLAAMITSAHNEKQLHCILIDESQFLSKEQVKQLTHVVDNLDIPVLCYGLKTDF QGELFSGSQYLLAWADKLVELKTICHCGRKANMVLRLDGSGKPMRDGEQVAIGGNESYES VCRKHFREFLWD ...
Generation of MT-III-deficient mice. The mouse MT-III gene was isolated from a 129Sv genomic library (Palmiter et al., 1992). A disruption vector was constructed by replacing a 2.2 kbXhoI-NdeI region containing the promoter and exons 1 and 2 with a neomycin resistance cassette and inserting thymidine kinase genes at BamHI and BstEII sites in the 5′ and 3′ flanking regions, respectively. Electroporation and selection of AB1 embryonic stem cells was performed as described (Thomas et al., 1995). Colonies were screened for targeting by PCR and confirmed by Southern blot analysis of DNA digested withSstI by using a 0.6 kb BstEII-NdeI fragment as a probe. Eight positive clones were identified from 840 clones screened. Four clones were used to generate chimeric mice as described (Thomas et al., 1995), but only one clone transmitted through the germline, and it was used to establish a line of mice carrying the disrupted allele. F2 and F3 generation C57Bl/129Sv hybrid mice of both sexes were used in ...
Rabbit polyclonal antibody raised against synthetic peptide of TK1. A synthetic peptide corresponding to N-terminus of human TK1. (PAB5614) - Products - Abnova
TY - CHAP. T1 - Osteonectin promoter-mediated suicide gene therapy of prostate cancer. AU - Hsiao, Wan Chi. AU - Sung, Shian Ying. AU - Chung, Leland W.K.. AU - Hsieh, Chia Ling. PY - 2019/1/1. Y1 - 2019/1/1. N2 - Suicide gene therapy using the herpes simplex virus thymidine kinase (HSV-tk) gene, combined with the prodrug ganciclovir (GCV) medication, is a promising approach for the treatment of malignant tumors, including prostate cancer. The success of this therapeutic strategy requires tissue- or tumor-specific gene expression and efficient gene delivery. In this chapter, we describe the experimental protocols of key methodologies, including promoter construction, reporter assay, adenoviral vector construction and preparation, HSV-tk enzymatic assay and cytotoxicity assay to evaluate the specificity and efficacy of osteonectin promoter-mediated HSV-tk/GCV suicide gene therapy of prostate cancer.. AB - Suicide gene therapy using the herpes simplex virus thymidine kinase (HSV-tk) gene, combined ...
TY - JOUR. T1 - Enhanced combined tumor-specific oncolysis and suicide gene therapy for prostate cancer using M6 promoter. AU - Ahn, M.. AU - Lee, S. J.. AU - Li, Xiaochun. AU - Jiménez, J. A.. AU - Zhang, Y. P.. AU - Bae, K. H.. AU - Mohammadi, Y.. AU - Kao, Chinghai. AU - Gardner, Thomas. PY - 2009/1. Y1 - 2009/1. N2 - Enzyme pro-drug suicide gene therapy has been hindered by inefficient viral delivery and gene transduction. To further explore the potential of this approach, we have developed AdIU1, a prostate-restricted replicative adenovirus (PRRA) armed with the herpes simplex virus thymidine kinase (HSV-TK). In our previous Ad-OC-TK/ACV phase I clinical trial, we demonstrated safety and proof of principle with a tissue-specific promoter-based TK/pro-drug therapy using a replication-defective adenovirus for the treatment of prostate cancer metastases. In this study, we aimed to inhibit the growth of androgen-independent (AI), PSA/PSMA-positive prostate cancer cells by AdIU1. In vitro the ...
We have developed a novel model for depleting mouse hepatic stellate cells (HSCs) that has allowed us to clarify their contributions to hepatic injury and fibrosis. Transgenic (Tg) mice expressing the herpes simplex virus thymidine kinase gene (HSV-Tk) driven by the mouse GFAP promoter were used to render proliferating HSCs susceptible to killing in response to ganciclovir (GCV). Effects of GCV were explored in primary HSCs and in vivo. Panlobular damage was provoked to maximize HSC depletion by combining CCl4 (centrilobular injury) with allyl alcohol (AA) (periportal injury), as well as in a bile duct ligation (BDL) model. Cell depletion in situ was quantified using dual immunofluorescence (IF) for desmin and GFAP. In primary HSCs isolated from both untreated wild-type (WT) and Tg mice, GCV induced cell death in ∼50% of HSCs from Tg, but not WT, mice. In TG mice treated with CCl4+AA+GCV, there was a significant decrease in GFAP and desmin-positive cells, compared to WT mice (∼65% reduction; ...
Acyclovir is a potent and selective antiviral agent. Unfortunately, drug-resistant (acyclovir-resistant) mutants have already been reported in herpes simplex virus type 1 (HSV-1) orofacial infections. We have developed a laboratory acyclovir-resistant HSV-1 mutant. The natural course of acyclovir-resistant HSV-1 keratitis was found to be less virulent than that observed in wild type HSV-1 keratitis, but the rate of ganglionic latency was as high as that induced by the parental strain. In vitro studies and in vivo observation of rabbit corneas infected with acyclovir-resistant HSV-1 both demonstrated a significant sensitivity to vidarabine and bromovinyldeoxyuridine ([E]-5-[2-bromovinyl]-2-deoxyuridine). The thymidine kinase activity of the acyclovir-resistant mutant was 69% of that of the wild type HSV-1 ...
The herpesvirus family comprises several widespread infectious pathogens. They infect a variety of animal hosts, including humans and cause complex clinical outcomes. Recently, the possible correlation between genital infection by human herpesviruses (HHVs) and male infertility has attracted considerable attention. In this chaper, we investigated the mechanism of HHV‐1‐induced infertility in transgenic (Tg) rats and its possible correlation with infertility in human males. Ectopic expression of HHV‐1 thymidine kinase (TK) in the testis of Tg rats increased male infertility. In addition, truncated TK proteins were found in postmeiotic spermatids of Tg rat testis, leading to progressive degeneration of germ cells and vacuolization of the seminiferous epithelium. These findings suggest the possibility that a similar process occurs within HHV‐infected human germ cells.
TY - JOUR. T1 - Ablation of Neurogenesis Attenuates Recovery of Motor Function after Focal Cerebral Ischemia in Middle-Aged Mice. AU - Sun, Fen. AU - Wang, Xiaomei. AU - Mao, Xiao Ou. AU - Xie, Lin. AU - Jin, Kunlin. PY - 2012/10/26. Y1 - 2012/10/26. N2 - Depletion of neurogenesis worsens functional outcome in young-adult mice after focal cerebral ischemia, but whether a similar effect occurs in older mice is unknown. Using middle-aged (12-month-old) transgenic (DCX-TK(+)) mice that express herpes simplex virus thymidine kinase (HSV-TK) under control of the doublecortin (DCX) promoter, we conditionally depleted DCX-positive cells in the subventricular zone (SVZ) and hippocampus by treatment with ganciclovir (GCV) for 14 days. Focal cerebral ischemia was induced by permanent occlusion of the middle cerebral artery (MCAO) or occlusion of the distal segment of middle cerebral artery (dMCAO) on day 14 of vehicle or GCV treatment and mice were killed 24 hr or 12 weeks later. Increased infarct volume ...
Objective: Neural stem cells (NSC) have an inherent brain tumor tropism that can be exploited for targeted delivery of therapeutic genes to invasive gliomas. Here, we demonstrate that the non-invasive intranasal administration of tumor-targeting NSC is able to deliver a novel suicide gene (TK007) to intracerebrally growing human glioblastoma xenografts.. Method: Murine NSC were genetically modified to express the novel herpes simplex virus thymidine kinase variant (TK007). The biological activity of the NSC-mediated TK007/ganciclovir (GCV) system was assessed in cell survival and bystander assays using various human glioma cell lines. Therapeutic effects of intratumoral (3x105 cells) and intranasal (1.5x106) NSC-TK007 application alone and the sequential combination of both was tested using an intracranial U87 human glioblastoma model in nude mice. All animals received 50 mg/kg GCV i.p. for five consecutive days. Two control groups received either NaCl instead of GCV or NSC containing the empty ...
Preferred Name: Valacyclovir Definition: The hydrochloride salt of the L-valyl ester of the antiviral drug acyclovir. Orally administered, valacyclovir is rapidly converted to acyclovir which inhibits viral DNA replication after further conversion to the nucleotide analog acyclovir triphosphate by viral thymidine kinase, cellular guanyl cyclase, and a number of other cellular enzymes. Acyclovir triphosphate competitively inhibits viral DNA polymerase; incorporates into and terminates the growing viral DNA chain; and inactivates viral DNA polymerase. The greater antiviral activity of acyclovir against herpes simplex virus (HSV) compared with varicella-zoster virus (VZV) is due to its more efficient phosphorylation by HSV thymidine kinase. NCI-GLOSS Definition: A substance that is being studied in the prevention of fungal, bacterial, and viral infections in patients undergoing donor stem cell transplantation with cells that are infected with cytomegalovirus. It belongs to the family of drugs ...
Glioblastoma is a highly aggressive malignant tumor involving glial cells in the human brain. We used high-throughput sequencing to comprehensively profile the small RNAs expressed in glioblastoma and non-tumor brain tissues. MicroRNAs (miRNAs) made up the large majority of small RNAs, and we identified over 400 different cellular pre-miRNAs. No known viral miRNAs were detected in any of the samples analyzed. Cluster analysis revealed several miRNAs that were significantly down-regulated in glioblastomas, including miR-128, miR-124, miR-7, miR-139, miR-95, and miR-873. Post-transcriptional editing was observed for several miRNAs, including the miR-376 family, miR-411, miR-381, and miR-379. Using the deep sequencing information, we designed a lentiviral vector expressing a cell suicide gene, the herpes simplex virus thymidine kinase (HSV-TK) gene, under the regulation of a miRNA, miR-128, that was found to be enriched in non-tumor brain tissue yet down-regulated in glioblastomas, Glioblastoma cells
Thymidine kinase (TK) belongs to the group of oncofetal enzymes. It catalyzes thymidine transformation to thymidine monophosphate in the presence of ATP. Elevated serum levels of TK can be found above all in the acute stages of malignant diseases of the hematopoietic system, in the bronchoendogenic carcinoma, carcinoma of the prostate, testis, and bladder.. ...
ID PCP1 preliminary; circular DNA; SYN; 3600 BP. XX AC S62800; ATCC37351; XX DT 01-JUL-1993 (Rel. 7, Created) DT 01-JUL-1995 (Rel. 12, Last updated, Version 1) XX DE Vertebrate/E.coli plasmid vector pCP1 - incomplete. XX KW cloning vector. XX OS Cloning vector OC Artificial sequences; Cloning vehicles. XX RN [1] RC plasmid from pUC9 & vaccinia virus 7.5kDa gene RC pCP1 from pCAT & plasmid RC pMM24 from pMM23 & SV40 72-bp repeats RA Cochran M.A., Mackett M., Moss B.; RT "Eukaryotic transient expression system dependent on transcription RT factors and regulatory DNA sequences of vaccinia virus"; RL Proc. Natl. Acad. Sci. U.S.A. 82:19-23(1985). XX RN [2] RC plasmid from pBR328 RC pGS8 from plasmid & vaccinia virus TK gene RC pMM1 from pUC9 & vaccinia virus TK gene RC pMM2 from pUC9 & vaccinia virus TK gene RC pMM3 from pMM1 & pMM2 RC pMM4 from pMM3 RC pMM5 from pMM4 RC pGS15 from pUC9 & pAG4 RC pGS19 from pGS15 & linker RC pGS20, pGS21 from pGS19 & pGS8 RC [pGS30 from cat gene] RC pCAT from pBR328 ...
Reliable markers for monitoring bladder tumor therapy are needed to evaluate treatment effectiveness. Thymidine kinase 1 (TK1) is an enzyme involved in DNA synthesis and therefore proliferation-dependent. Serum concentration of TK1 (STK1) correlates with malignancy in various types of cancer, thus reflecting treatment results. This study explores for the first time the use of STK1 concentration, both as a prognostic marker and to monitor the outcome of bladder carcinoma surgery. STK1 in 56 bladder carcinoma patients was measured pre-operatively, and post-operatively at 1 week and 1, 3, and 6 months, using an immune ECL dot blot assay. An anti-TK1 chicken IgY antibody was used to determine STK1 concentrations. Mean pre-operative STK1 of bladder carcinoma patients was significantly higher than that of healthy individuals, with no overlap of individual values. STK1 concentrations increased significantly with tumor stage (I-III) and T-values (T1-T2), but not tumor grade (G1-G4). STK1 gradually ...
In the stab-injured spinal cord of GFAP-TK mice not given GCV, we found that all TK-expressing cells also expressed GFAP, and that ,98%, but not all, of GFAP-expressing reactive astrocytes also expressed detectable levels of transgene-derived TK (Fig. 2A-D,G). The inability to detect TK in all GFAP-expressing cells may be attributable to the discontinuous activity of the GFAP gene and differences in the intracellular half-life of the GFAP and TK proteins, as discussed elsewhere (Bush et al., 1999; Imura et al., 2003). We found no evidence for TK expression by cells that did not express GFAP.. To evaluate the degree of astrocyte cell division induced by SCI, we administered BrdU during the first 4 d after injury (Fig. 1A). At 14 d after injury, control mice exhibited many BrdU-labeled cells in tissue immediately adjacent to the stab wound (Fig. 2E,F); most of these BrdU-labeled cells expressed GFAP (Fig. 2H) and TK.. There was no qualitatively detectable difference in the response to SCI in all ...
AVIRAX Fabrigen Acyclovir Antiviral Action And Clinical Pharmacology: Acyclovir, a synthetic acyclic nucleoside analog, is a substrate with a high degree of specificity for herpes simplex and varicella-zoster specified thymidine kinase. Acyclovir is a poor substrate for host cell-specified thymidine kinase. Herpes simplex and varicella-zoster specified thymidine kinase transform acyclovir to its monophosphate which is then transformed by a number of cellular enzymes to acyclovir diphosphate and acyclovir triphosphate. Acyclovir triphosphate is both an inhibitor of, and a substrate for, herpesvirus-specified DNA polymerase. Although the cellular a-DNA polymerase in infected cells may also be inhibited by acyclovir triphosphate, this occurs only at concentrations of acyclovir triphosphate which are higher than those which inhibit the herpesvirus-specified DNA polymerase. Acyclovir is selectively converted to its active form in herpesvirus-infected cells and is thus preferentially taken up by these ...
The nucleotide sequence of the UL31 open reading frame is predicted to encode a basic protein with a hydrophilic amino terminus and a nuclear localization signal. To identify its gene product, we constructed a viral genome in which the thymidine kinase gene was inserted between the UL31 and UL32 open reading frames. The thymidine kinase gene was then deleted, and in the process, the 5 terminus of the UL31 open reading frame was replaced with a 64-bp sequence in frame with the complete, authentic sequence of the UL31 open reading frame. The inserted sequence encoded a hydrophilic epitope derived from glycoprotein B of human cytomegalovirus and for which a monoclonal antibody is available. We report that in infected cells, the tagged protein localized in and was dispersed throughout the nucleus. Nuclear fractionation studies revealed that the UL31 protein partitions with the nuclear matrix. The protein is phosphorylated in infected cells maintained in medium containing 32Pi. ...
A valid GLP study was performed to investigate the potential of the test item to induce mutations at the mouse lymphoma thymidine kinase locus using the cell line L5178Y. The method followed was that described in OECD TG 476. The assay was performed in three independent experiments, using two parallel cultures each. Experiment I, and II were performed with and without liver microsomal activation and a treatment period of 4 hours. The supplementary experiment III was solely performed with metabolic activation (4 hours treatment) to cover the cytotoxic range of approximately 10- 20% RTG that was not covered in the second experiment with metabolic activation. The concentration range of the main experiments was limited by the solubility of the test item in aqueous media and by cytotoxic effects. Relevant cytotoxic effects occurred at 40 µg/mL in the first experiment without metabolic activation and at 80.0 µg/mL in the presence of metabolic activation. In the second experiment cytotoxicity was ...