Division of labor within the DNA damage tolerance system reveals non-epistatic and clinically actionable targets for precision cancer medicine

Crosslink repair depends on the Fanconi anemia pathway and translesion synthesis polymerases that replicate over unhooked crosslinks. Translesion synthesis is regulated via ubiquitination of PCNA, and independently via translesion synthesis polymerase REV1. The division of labor between PCNA-ubiquitination and REV1 in interstrand crosslink repair is unclear. Inhibition of either of these pathways has been proposed as a strategy to increase cytotoxicity of platinating agents in cancer treatment. Here, we defined the importance of PCNA-ubiquitination and REV1 for DNA in mammalian ICL repair. In mice, loss of PCNA-ubiquitination, but not REV1, resulted in germ cell defects and hypersensitivity to cisplatin. Loss of PCNA-ubiquitination, but not REV1 sensitized mammalian cancer cell lines to cisplatin. We identify polymerase Kappa as essential in tolerating DNA damage-induced lesions, in particular cisplatin lesions. Polk-deficient tumors were controlled by cisplatin treatment and it significantly delayed tumor outgrowth and increased overall survival of tumor bearing mice. Our results indicate that PCNA-ubiquitination and REV1 play distinct roles in DNA damage tolerance. Moreover, our results highlight POLK as a critical TLS polymerase in tolerating multiple genotoxic lesions, including cisplatin lesions. The relative frequent loss of Polk in cancers indicates an exploitable vulnerability for precision cancer medicine.     

Isolation and characterization of a virulent bacteriophage from Staphylococcus carnosus

Abstract A virulent bacteriophage, øSK311, was isolated from Staphylococcus carnosus , an organism used as a starter culture for the production of dry sausage. Electron microscopic studies revealed that this bacteriophage showed some morphological similarities with the Escherichia coli phages T4 and λ. The host range of øSK311 extends over 9 different staphylococcal species. A phage-resistant mutant of S. carnosus could be isolated. Cells of this mutant exhibited a capsule-like structure. The DNA of øSK311 possesses a G + C content of 31.4 mol% and appears to be highly modified.     

The chimeric cytokine Hyper-IL-6 enhances the efficiency of lentiviral gene transfer in hepatocytes both in vitro and in vivo

Lentiviral vectors have been used for gene transfer into the liver but their ability to efficiently transduce quiescent hepatocytes remains controversial. Lentivirus-mediated gene transfer is more efficient in cycling cells. We determine the effect of H-IL6 in the lentiviral transduction. The lentiviral vector was used to transduce HepG2 cells and mice liver cells, previously treated with H-IL6. The highest transduction level was observed in HepG2 cells treated with 30 ng/mL H-IL6 and in the mice that received 4 μg H-IL6. Our results suggest that H-IL6 is an inducer of lentiviral gene transfer into the liver cells without any toxicity.     

Control of Liriomyza langei on chrysanthemum by Diglyphus isaea produced with a standard or modified parasitoid rearing technique

Abstract: Overproduction of male parasitoids during mass rearing will increase costs for biological control because wasp shipments contain fewer females and only females kill hosts directly. We have developed a rearing technique capable of significantly reducing male‐biased sex ratios in Diglyphus isaea (Walker) (Hym., Eulophidae), a commercially reared parasitoid of agromyzid leafminers. In this study, we examined the effect of rearing technique on the efficacy of D. isaea for biological control of Liriomyza langei Frick (Dip., Agromyzidae) on chrysanthemum, Dendranthema grandiflora Tzvelev var. ‘Miramar’. We produced D. isaea on mixtures of small and large hosts (our modified technique) or on only large hosts (simulating commercial mass‐rearing) and compared: (1) control of L. langei with D. isaea produced by the two rearing techniques, and (2) damage and yield of unprotected and protected plants. The two rearing techniques produced similar numbers of waSPS per rearing cohort, but the ‘modified’ technique reduced the proportion of males by approximately 13%. The two techniques also produced females of similar size, but the ‘modified’ technique produced smaller males. In greenhouse trials simulating leafminer infestations of potted chrysanthemums during commercial production, we found no significant differences between the levels of control obtained by releasing identical numbers and sex ratios of adult waSPS produced by either rearing technique. Mine counts on plants protected by waSPS of either rearing history were similar and around 30–70% less than unprotected plants during most of the 11‐week crop cycle. At crop harvest, more than half of the foliage on protected plants was undamaged compared with <10% on unprotected crops. Damage to the flower stems of protected plants was relatively light in the top half of the canopy compared with the bottom half. Protected plants were around 10–15% taller and produced twice as many flower buds compared with unprotected plants. Our ‘modified’ rearing technique can reduce overproduction of males in D. isaea with no compromise in biological control efficacy. Adoption of our rearing technique by commercial insectaries could reduce implementation costs for not only D. isaea but also other parasitoids that show host‐size‐dependent sex allocation.     

Influenza epidemics in the USSR and Czechoslovakia in 1979-1984

In this work materials characterizing the appearance and development of influenza epidemic at the territories of the USSR and the Czech Socialist Republic are presented, the common features and differences of the epidemic process in both countries are recorded. The work shows that in both countries the appearance of this epidemic is caused by the same virus. In most cases the epidemic started earlier and lasted longer in the USSR, but morbidity rate during the epidemic was, on the whole, higher in the Czech Socialist Republic. Similarity in the course of the primary period of the epidemic processes from their appearance to their maximum rise was observed. In both countries the maximum rise of morbidity rate was registered on weeks 3-4 from the beginning of the epidemic.     

DFP-sensitive polypeptides of the guinea pig peritoneal macrophage

Guinea pig peritoneal exudate macrophages were reacted with [³H] diisopropyl fluorophosphate (10^?7 to 10^?4 M) and subjected to sodium dodecyl sulfate-polyacrylamide disc electrophoresis and fluorography. Macrophages reacted with 10^?5 M [³H]diisopropyl fluorophosphate contain eight major ³H-labelled polypeptides which have apparent molecular weights of 83,000, 75,000, 63,000, 48,000, 41,000, 30,000, 26,000, and 25,000. The sensitive polypeptides were not seen when guinea pig polymorphonuclear leukocytes, lymph node lymphocytes, erythrocytes, serum or plasma were reacted with [³H]diisopropyl fluorophosphate, demonstrating that these components are particular to the macrophage. The finding of a large number of diisopropyl fluorophosphate-sensitive proteins associated exclusively with the macrophage supports the concept that serine esterases play a unique role in macrophage physiology.     

A new spectrophotometric assay for enzymes of purine metabolism. II. Determination of guanase activity.

A new method for the determination of guanase is described. Xanthine, the product of the guanase reaction, is oxidized by xanthine oxidase, forming uric acid and hydrogen peroxide. Hydrogen peroxide is further reduced to water by catalase in the presence of ethanol. The acetaldehyde formed in this reaction step is dehydrogenated NAD or NADP dependent by aldehyde dehydrogenase. The NADH or NADPH production is measured and utilized for the calculation of the guanase activity. The sensitivity of the method can be doubled by the addition of uricase, which oxidizes uric acid to permit the formation of another mole of hydrogen peroxide.