Efficient production of infectious viruses requires enzymatic activity of Epstein-Barr virus protein kinase

AbstractThe Epstein-Barr virus (EBV) BGLF4 gene product is the only protein kinase encoded by the virus genome. In order to elucidate its physiological roles in viral productive replication, we here established a BGLF4-knockout mutant and a revertant virus. While the levels of viral DNA replication of the deficient mutant were equivalent to those of the wild-type and the revertant, virus production was significantly impaired. Expression of the BGLF4 protein in trans fully complemented the low yield of the mutant virus, while expression of a kinase-dead (K102I) form of the protein failed to restore the virus titer. These results demonstrate that BGLF4 plays a significant role in production of infectious viruses and that the kinase activity is crucial

遺伝子診断教育のための簡便な毛髪形態（EDAR）及び耳垢型（ABCC11）遺伝子多型解析法

Recently, progress in molecular biology is being made in various fields such as biology, agriculture and medical and pharmaceutical sciences. Students graduating from not only pharmaceutical departments but also general high schools will be required to have knowledge of molecular biology techniques for understanding prevailing technologies. However, it is difficult to include such experiments in a curriculum for students at general high schools and universities because of safety aspect, expensive materials and experimental equipment and time consumed. This report introduces the convenient experimental methods on human DNA polymorphisms for the genotyping of the hair thickness-determining gene EDAR on the chromosome 2 and the earwax type gene ABCC11 on the chromosome 16 using a PCR-RFLP method. This experiment enables each student to handle DNA safely. The total experiment time is less than 6 hours. The PCR-RFLP experiment proposed here is a suitable genotyping method to acquire molecular biological knowledge and techniques

Chemical synthesis of oligodeoxyribonucleotides containing the Dewar valence isomer of the (6–4) photoproduct and their use in (6–4) photolyase studies

The pyrimidine(6–4)pyrimidone photoproduct, a major UV lesion formed between adjacent pyrimidine bases, is transformed to its Dewar valence isomer upon exposure to UVA/UVB light. We have synthesized a phosphoramidite building block of the Dewar photoproduct formed at the thymidylyl(3′–5′)thymidine site and incorporated it into oligodeoxyribonucleotides. The diastereoisomers of the partially protected dinucleoside monophosphate bearing the (6–4) photoproduct, which were caused by the chirality of the phosphorus atom, were separated by reversed-phase chromatography, and the (6–4) photoproduct was converted to the Dewar photoproduct by irradiation of each isomer with Pyrex-filtered light from a high-pressure mercury lamp. The Dewar photoproduct was stable under both acidic and alkaline conditions at room temperature. After characterization of the isomerized base moiety by NMR spectroscopy, a phosphoramidite building block was synthesized in three steps. Although the ordinary method could be used for the oligonucleotide synthesis, benzimidazolium triflate as an alternative activator yielded better results. The oligonucleotides were used for the analysis of the reaction and the binding of Xenopus (6–4) photolyase. Although the affinity of this enzyme for the Dewar photoproduct-containing duplex was reportedly similar to that for the (6–4) photoproduct-containing substrate, the results suggested a difference in the binding mode

Subserosal Inflammatory Pseudotumor Causing Intussusception

We present an adult case of intussusception caused by subserosal inflammatory pseudotumor of the terminal ileum. Enteric intussusception was diagnosed by characteristic Xray finding, so called "beak-like" filling defect in the terminal ileum. An exploratory laparotomy revealed focal subserosal thickening in the terminal ileum, and partial ileocolectomy was performed. Microscopically, we identified a pseudotumor that extended through the muscularis propria into serosa. The pseudotumor was composed of fibrous stroma and chronic inflammatory cells with calcification

唾液中カフェイン薬物動態に薬物代謝酵素CYP1A2 遺伝子多型が及ぼす影響に関する研究

Though drugstore pharmacists can also be known as primary health care pharmacists, it cannot yet be said that they completely fill this role. We believe that it is crucial for pharmacists to consult reliably with patients. To develop this ability, it is necessary to select a familiar and relevant theme by considering various factors that are worth discussing in their own right through conducting simple verification experiments. Caffeine is found in many foods and drinks, as well as in over the counter cold remedies and analgesics. Moreover, it has been widely used around the world to probe the effectiveness of drug-metabolizing enzyme activity of medications for liver disease. We have built a simple and easy experiment system to evaluate the pharmacokinetics of caffeine in saliva, and we examined the relationship between the drug-metabolizing enzyme CYP1A2 genetic polymorphism and various related constitutional hereditary factors

Photosensitized [2 + 2] cycloaddition of N-acetylated cytosine affords stereoselective formation of cyclobutane pyrimidine dimer

Photocycloaddition between two adjacent bases in DNA produces a cyclobutane pyrimidine dimer (CPD), which is one of the major UV-induced DNA lesions, with either the cis-syn or trans-syn structure. In this study, we investigated the photosensitized intramolecular cycloaddition of partially-protected thymidylyl-(3′→5′)-N4-acetyl-2′-deoxy-5-methylcytidine, to clarify the effect of the base modification on the cycloaddition reaction. The reaction resulted in the stereoselective formation of the trans-syn CPD, followed by hydrolysis of the acetylamino group. The same result was obtained for the photocycloaddition of thymidylyl-(3′→5′)-N4-acetyl-2′-deoxycytidine, whereas both the cis-syn and trans-syn CPDs were formed from thymidylyl-(3′→5′)-thymidine. Kinetic analyses revealed that the activation energy of the acid-catalyzed hydrolysis is comparable to that reported for the thymine-cytosine CPD. These findings provided a new strategy for the synthesis of oligonucleotides containing the trans-syn CPD. Using the synthesized oligonucleotide, translesion synthesis by human DNA polymerase η was analyzed