PURPOSE: 5-iododeoxyuridine analogues have been exclusively developed for the potential antiviral and antitumor therapeutic agents. In this study, we synthesized carbocyclic radioiododeoxyuridineanalogue (ddIVDU) and carbocyclic intermediate as efficient carbocyclic radiopharmaceuticals. MATERIALS AND METHODS: The synthesis is LAH reduction, hetero Diels-Alder reaction as key reactions including Pd(0)-catalyzed coupling reaction together with organotin. MCA-RH7777 (MCA) and MCA-tk (HSV1-tk positive) cells were treated with various concentration of carbocyclic ddIVDU, and GCV. Cytotoxicity was measured by the MTS methods. For in vitro uptake study, MCA and MCA-tk cells were incubated with 1uCi of [(125)I]carbocyclic ddIVDU. Accumulated radioactivity was measured after various incubation times. RESULTS: The synthesis of ddIVDU and precursor for radioiodination were achieved from cyclopentadiene in good overall yield, respectively. The radioiododemetallation for radiolabeling gave more than 80% yield with > 95% radiochemical purity. GCV was more toxic than carbocyclic ddIVDU in MCA-tk cells. Accumulation of [(125)I]carbocyclic ddIVDU was higher in MCA-tk cells than MCA cells. CONCLUSION: Biological data reveal that ddIVDU is stable in vitro, less toxic than ganciclovir (GCV), and selective in HSV1-tk expressed cells. Thus, this new carbocyclic nucleoside, referred to in this paper as carbocyclic 2',3'-didehydro-2',3'-dideoxy-5- iodovinyluridine (carbocyclic ddIVDU), is a potential imaging probe for HSV1-tk.
Cycloaddition Reaction ; Ganciclovir ; Herpesvirus 1, Human ; Idoxuridine ; Radioactivity ; Radiopharmaceuticals

Background : Gene transfer with vectors derived from murine retroviruses is restricted to cells which are proliferating and synthesizing DNA at the time of infection. Accordingly, selective introduction of genes encoding for susceptibility to otherwise nontoxic drugs (suicide genes) into proliferating tumor may be used to treat cancer. We investigated the efficacy of in vitro transduction of neuroblastoma cell with the herpes simplex-thymidine kinase (HSV-tk) gene followed by administration of the antiviral drug ganciclovir. METHODS: The LNC/tK vector was transfered in vitro into mouse Neuro 2a cell lines (ATCC) and the transduced cell lines were selected in G-418, 500microgram/ml, for 14 days. Onex104 cells were cultured in 96 well culture plates in increasing concentrations of ganciclovir for 72 hours. The sesitivity to ganciclivir of these HSV-tk transduced, G-418 selected cells was measured with MTT assay RESULTS: The survival of HSV-tk transduced 1x104 neuro 2a cell lines is 103+/-3.5%, 68+/-4.2%, 54+/-3.8%, 17+/-2.6%, 13+/-3.1% at the concentration of 0, 0.1, 1.0, 10, 20microgram/ml ganciclovir, respectively. And the survival of HSV-tk not transduced 1x104 neuro 2a cell lines is 100+/-4.5%, 97+/-5.6%, 104+/-3.5%, 106+/-3.8%, 101+/-4.2%. CONCLUSION: We concluded that in vitro transduction of neuroblastoma cell with the herpes simplex-thymidine kinase gene followed by administration of the antiviral drug ganciclovir is very effective.
Animals ; Cell Line* ; DNA ; Ganciclovir ; Mice ; Neuroblastoma* ; Phosphotransferases ; Retroviridae ; Zidovudine*

Background : Gene transfer with vectors derived from murine retroviruses is restricted to cells which are proliferating and synthesizing DNA at the time of infection. Accordingly, selective introduction of genes encoding for susceptibility to otherwise nontoxic drugs (suicide genes) into proliferating tumor may be used to treat cancer. We investigated the efficacy of in vitro transduction of neuroblastoma cell with the herpes simplex-thymidine kinase (HSV-tk) gene followed by administration of the antiviral drug ganciclovir. METHODS: The LNC/tK vector was transfered in vitro into mouse Neuro 2a cell lines (ATCC) and the transduced cell lines were selected in G-418, 500microgram/ml, for 14 days. Onex104 cells were cultured in 96 well culture plates in increasing concentrations of ganciclovir for 72 hours. The sesitivity to ganciclivir of these HSV-tk transduced, G-418 selected cells was measured with MTT assay RESULTS: The survival of HSV-tk transduced 1x104 neuro 2a cell lines is 103+/-3.5%, 68+/-4.2%, 54+/-3.8%, 17+/-2.6%, 13+/-3.1% at the concentration of 0, 0.1, 1.0, 10, 20microgram/ml ganciclovir, respectively. And the survival of HSV-tk not transduced 1x104 neuro 2a cell lines is 100+/-4.5%, 97+/-5.6%, 104+/-3.5%, 106+/-3.8%, 101+/-4.2%. CONCLUSION: We concluded that in vitro transduction of neuroblastoma cell with the herpes simplex-thymidine kinase gene followed by administration of the antiviral drug ganciclovir is very effective.
Animals ; Cell Line* ; DNA ; Ganciclovir ; Mice ; Neuroblastoma* ; Phosphotransferases ; Retroviridae ; Zidovudine*

It has been suspected that various infections, including cytomegalovirus(CMV) infection, are associated with IgA nephropathy. In case of CMV infection, ganciclovir is known to be a treatment of choice for severe CMV infection in general. But ganciclovir has a lot of severe toxicity, so children with normal immunity are seldom treated by ganciclovir when CMV infection is suspected. On the other hand, intravenous immunoglobulin can also be used to treat CMV infection.
Child ; Cytomegalovirus ; Ganciclovir ; Glomerulonephritis, IGA ; Hand ; Humans ; Immunoglobulin A ; Immunoglobulins ; Pregnenediones

Congenital cytomegalovirus (CMV) infection is the leading identified nongenetic cause of congenital sensorineural hearing loss (SNHL). In symptomatic CMV infected infants, systemic ganciclovir therapy prevents hearing deterioration or maintains normal hearing in early childhood. In this article, we present a case of hearing improvement in a congenital CMV infected infant with SNHL.
Cytomegalovirus ; Ganciclovir ; Hearing ; Hearing Loss ; Hearing Loss, Sensorineural ; Humans ; Infant

OBJECTIVETo detect whether the murine T-cell lymphoma cell line EL4 could be infected by murine cytomegalovirus (MCMV), and to observe the morphological changes and apoptosis of El4 cells before and after infection.

METHODSEL4 cells were infected with MCMV smith strain with 1, 10 and 100 multiplicity of infection (MOI) respectively. The morphology of the cells was observed by light microscopy and Wright's-Giemsa staining. The survival rate was calculated by trypan blue staining. RT-PCR-based assay was used to detect the copy number of MCMV-DNA in the infected ELA cells. Flow cytometry was used to detect apoptosis. RT-qPCR was used to detect the mRNA expression levels of P53, P21, cFlip and Caspase 8. The protein expression levels of Caspase8, P53, BAX, BCL-2 and Cleaved Caspase3 proteins were detected by Western blot.

RESULTSAfter Wright-Giemsa staining, it was found that the infected EL4 cells displayed larger volume, irregular nuclei and the folded twist under light microscopy. Compared with the normal control group, the survival rate of EL4 cells decreased, and the apoptosis rate statistically significantly (P<0.05) increased with the increasing MOI and the infected time in each group. While, the level of apoptosis protein P53, BAX/BCL-2, Cleaved-caspase3 and Caspase8 were up-regulated. And the survival rate, apoptosis rate and the apoptosis protein level of infected EL4 cells with MOI=10 were the most obvious at the 5day. Compared with MCMV infection group (MOI=60), the content of MCMV DNA in EL4 cells was decreased in MCMV+GGV group ［MOI=60, GCV 25 (g/ml)］, and the cell apoptosis rate and apoptosis protein expression of P53, Caspase8, BAX/BCL-2 were decreased (P<0.05).

CONCLUSIONMurine T-cell lymphoma cell line EL4 can be infected by MCMV and displayes an obvious apoptosis phenomenon. MCMV may up-regulate the expression levels of apoptosis protein P53, BAX-BCL-2, Cleaved-caspase3 and Caspase8 in EL4 cells. The drug ganciclovir reduces the copy mumber of MCMV DNA in infected EL4 cells and inhibited the killing effect of MCMV on EL4 cells.

BACKGROUND: This study was performed to determine the cut-off value and the predictability of symptomatic human cytomegalovirus (HCMV) infection according to the peak value of HCMV antigenemia assay in kidney transplant recipients. MATERIALS AND METHODS: We reviewed the results of HCMV antigenemia assay (Chemicon, CA, USA) in patients who received kidney transplantation at our institution from May 2003 through May 2008, and investigated the existence and the type of HCMV infection by the medical record review. Patients who underwent the test only once during the episode or those who received ganciclovir for more than 48hrs before the test were excluded. The receiver-operator characteristic curve was drawn and the point showing maximum likelihood ratio (LR) was chosen as the cut-off value of symptomatic HCMV infection. RESULTS: A total of 689 episodes were screened and 134 episodes were enrolled. Thirty-three (24.6%) episodes were symptomatic HCMV infection, 23 (17.2%) episodes were associated with HCMV syndrome, and 10 (7.5%) episodes were tissue-invasive diseases. The maximum LR was 7.5 (95% confidence interval, 4.014.2) and the cut-off value was 29.5 cells/200,000 WBC. The sensitivity, specificity, positive predictive value, and negative predictive value were 66.7%, 91.1%, 71.0%, and 89.3%, respectively. CONCLUSIONS: The cut-off value of symptomatic HCMV infection by the peak value of HCMV antigenemia assay in our study was similar with previous results, although the sensitivity was relatively low.
Cytomegalovirus ; Ganciclovir ; Humans ; Kidney ; Kidney Transplantation ; Medical Records ; Sensitivity and Specificity

BACKGROUND: This study was performed to determine the cut-off value and the predictability of symptomatic human cytomegalovirus (HCMV) infection according to the peak value of HCMV antigenemia assay in kidney transplant recipients. MATERIALS AND METHODS: We reviewed the results of HCMV antigenemia assay (Chemicon, CA, USA) in patients who received kidney transplantation at our institution from May 2003 through May 2008, and investigated the existence and the type of HCMV infection by the medical record review. Patients who underwent the test only once during the episode or those who received ganciclovir for more than 48hrs before the test were excluded. The receiver-operator characteristic curve was drawn and the point showing maximum likelihood ratio (LR) was chosen as the cut-off value of symptomatic HCMV infection. RESULTS: A total of 689 episodes were screened and 134 episodes were enrolled. Thirty-three (24.6%) episodes were symptomatic HCMV infection, 23 (17.2%) episodes were associated with HCMV syndrome, and 10 (7.5%) episodes were tissue-invasive diseases. The maximum LR was 7.5 (95% confidence interval, 4.014.2) and the cut-off value was 29.5 cells/200,000 WBC. The sensitivity, specificity, positive predictive value, and negative predictive value were 66.7%, 91.1%, 71.0%, and 89.3%, respectively. CONCLUSIONS: The cut-off value of symptomatic HCMV infection by the peak value of HCMV antigenemia assay in our study was similar with previous results, although the sensitivity was relatively low.
Cytomegalovirus ; Ganciclovir ; Humans ; Kidney ; Kidney Transplantation ; Medical Records ; Sensitivity and Specificity

OBJECTIVETo evaluate the method of adenovirus vector-mediated herpes simplex virus-thymidine kinase gene (ADV-TK)/ganciclovir(GCV) system and photodynamic therapy(PDT) for treating the oral malignant tumor.

METHODSTen patients who were suffering from oral malignant tumor of the different positions were selected, and injected with the hematoporphyrin derivative (HPD), irradiated by picking the corresponding laser frequency, and injected the ADV-TK gene into the tumor body and periphery. By the imaging and the hemodynamics analysis, the clinical efficacy after infusing vein with GCV was assessed.

RESULTSAfter the combination method treatments, patients' tumors appeared clear shrinkage or complete extinction. There was obvious fall of the blood flow volume in the tumor body. Imaging results showed significantly differences. So it was a perfect and effective treatment.

CONCLUSIONThere are many advantages to apply the ADV-TK/GCV system and PDT treatment on the oral malignant tumor, minimal side effects and greater clinical security. It is a safe and credible therapy which can be offered for curing the oral malignant tumor systematically.

Human cytomegalovirus encodes an unusual protein kinase UL97 which can phosphorylate exogenous substrates, including histone H2B and nucleoside analogs such as ganciclovir. The previous result interestingly showed that the peptides phosphorylated by UL97 have K/R at the 5 positions (P+5) downstream from the pSer. To confirm the importance of the basic residue in the position, we used two peptide libraries, 4S4K (MAXXXXSXXXXKXANNN) and 4S6N (MAXXXXSXXXXXXNNN). The activity of phosphorylation by UL97 was higher in the peptide library 4S4K than 4S6N, suggesting the importance of basic residue at P+5 position. The screening with a peptide library 4S4K showed slight tendencies for N in the P+1 and P+2, M in the P+2, K in the P+4 and P+6 positions and several amino acids in the other positions. This result will give information to develop an optimal peptide for screening a novel UL97 inhibitor.
Amino Acids ; Cytomegalovirus* ; Ganciclovir ; Histones ; Humans* ; Mass Screening* ; Peptide Library* ; Peptides ; Phosphorylation ; Protein Kinases* ; Substrate Specificity*