XR-C1, a new CHO cell mutant which is defective in DNA-PKcs, is impaired in both V(D)J coding and signal joint formation

DNA-dependent protein kinase (DNA-PK) plays an important role in DNA double-strand break (DSB) repair and V(D)J recombination. We have isolated a new X-ray-sensitive CHO cell line, XR-C1, which is impaired in DSB repair and which was assigned to complementation group 7, the group that is defective in the XRCC7 / SCID ( Prkdc ) gene encoding the catalytic subunit of DNA-PK (DNA-PKcs). Consistent with this complementation analysis, XR-C1 cells lackeddetectable DNA-PKcs protein, did not display DNA-PK catalytic activity and were complemented by the introduction of a single human chromosome 8 (providing the Prkdc gene). The impact of the XR-C1 mutation on V(D)J recombination was quite different from that found in most rodent cells defective in DNA-PKcs, which are preferentially blocked in coding joint formation, whereas XR-C1 cells were defective in forming both coding and signal joints. These results suggest that DNA-PKcs is required for both coding and signal joint formation during V(D)J recombination and that the XR-C1 mutant cell line may prove to be a useful tool in understanding this pathway.     

The univector plasmid-fusion system, a method for rapid construction of recombinant DNA without restriction enzymes

BACKGROUND:. Modern biological research is highly dependent upon recombinant DNA technology. Conventional cloning methods are time-consuming and lack uniformity. Thus, biological research is in great need of new techniques to rapidly, systematically and uniformly manipulate the large sets of genes currently available from genome projects. RESULTS:. We describe a series of new cloning methods that facilitate the rapid and systematic construction of recombinant DNA molecules. The central cloning method is named the univector plasmid-fusion system (UPS). The UPS uses Cre-lox site-specific recombination to catalyze plasmid fusion between the univector - a plasmid containing the gene of interest - and host vectors containing regulatory information. Fusion events are genetically selected and place the gene under the control of new regulatory elements. A second UPS-related method allows for the precise transfer of coding sequences only from the univector into a host vector. The UPS eliminates the need for restriction enzymes, DNA ligases and many in vitro manipulations required for subcloning, and allows for the rapid construction of multiple constructs for expression in multiple organisms. We demonstrate that UPS can also be used to transfer whole libraries into new vectors. Additional adaptations are described, including directional PCR cloning and the generation of 3' end gene fusions using homologous recombination in Escherichia coli. CONCLUSIONS:. Together, these recombination-based cloning methods constitute a new comprehensive approach for the rapid and efficient generation of recombinant DNA that can be used for parallel processing of large gene sets, a feature that will facilitate future genomic analysis.     

Cognitive–behavior therapy versus pharmacotherapy: Now that the jury's returned its verdict, it's time to present the rest of the evidence.

Recent reanalyses suggest that pharmacotherapy was superior to cognitive–behavior therapy in the acute treatment of more severely depressed outpatients in the National Institute of Mental Health Treatment of Depression Collaborative Research Program (TDCRP). At the same time, this finding was neither robust across sites within the TDCRP nor consistent with findings from other studies. D. F. Klein (1990) has argued that those other studies were inherently flawed because they did not include pill-placebo controls, an argument that he extended to drug–psychotherapy comparisons in the treatment of panic as well. It is agreed that the inclusion of such controls would have facilitated the interpretation of the findings, but it is not agreed that their omission rendered those studies uninterpretable. Cognitive–behavior therapy remains a viable alternative to pharmacotherapy in the treatment of depression and a particularly promising intervention in the treatment of panic disorder. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Prospects for future comparisons between drugs and psychotherapy: Lessons from the CBT-versus-pharmacotherapy exchange.

In these concluding comments to the exchange among N. S. Jacobson and S. D. Hollon, D. F. Klein, I. Elkin et al, and R. J. McNally (see record 1996-02770-007, 1996-02770-008, 1996-02770-010, and 1996-02770-009, respectively) issues of continuing controversy are highlighted: the best ways to control for the allegiance effect, the complexities of assessing competence, and the role of pill placebos in comparisons between cognitive–behavior therapy (CBT) and pharmacotherapy. It is concluded that controlling for allegiance within site is the optimal strategy, that measures of competence must take context into account, and that pill placebo controls are optimal but not essential to comparisons between CBT and pharmacotherapy. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Effects of dietary vegetable oils on behavior and drug responses in mice

Previously, we noted significant differences in the behavioral patterns of mice fed safflower oil with a very low alpha-linolenate/linoleate ratio and perilla oil with a high alpha-linolenate/linoleate ratio from mothers to offsprings. In this report, we compared the behavior and drug responses in mice fed diets containing six different vegetable oils-corn, rapeseed, soybean, safflower, perilla and a mixture of perilla and safflower oils- for a relatively short period: 8 months after weaning. Soybean oil is a component of most conventional diets and was used as a control. The alpha-linolenate/linoleate ratios of the oils appeared to affect the locomotor activities in a wheel cage: the activity decreased in the order of safflower, the mixture (1:1) and the perilla oil groups. However, the rapeseed oil group exhibited much higher locomotor activity than that expected from the alpha-linolenate/linoleate ratio. Additionally, the rapeseed oil group exhibited unusual behavior patterns, including higher ambulation and rearing activities, faster acquisition of the water maze task and slower habituation behavior as compared with the control group. Susceptibility to pentobarbital anesthesia tended to be higher in the rapeseed oil group. The differences in the alpha-linolenate/linoleate ratios of these oils alone do not account for the observed differences in the behavioral patterns among the six dietary groups. Although we cannot exclude the possibility that the observed behavioral anomaly is due to the unique fatty acid composition of rapeseed oil, we speculate that a factor(s) other than fatty acids in rapeseed oil affected nervous system functions.     

Intraoperative monitoring with stimulus-evoked electromyography during placement of iliosacral screws. An initial clinical study

Cells of the innate immune system secrete cytokines early in immune responses that guide maturing T helper (Th) cells along appropriate lineages. This study investigates the role of cytokine networks, bridging the innate and acquired immune systems, in the pathogenesis of an organ specific autoimmune disease. Experimental allergic encephalomyelitis (EAE), a demyelinating disease of the central nervous system, is widely used as an animal model for multiple sclerosis. We demonstrate that interleukin (IL)-12 is essential for the generation of the autoreactive Th1 cells that induce EAE, both in the presence and absence of interferon gamma. The disease-promoting effects of IL-12 are antagonized by IL-10 produced by an antigen nonspecific CD4+ T cell which, in turn, is regulated by the endogenous production of IL-12. This unique immunoregulatory circuit appears to play a critical role in controlling Th cell differentiation and provides a mechanism by which microbial triggers of the innate immune system can modulate autoimmune disease.     

Europa's differentiated internal structure: inferences from four Galileo encounters

Radio Doppler data from four encounters of the Galileo spacecraft with the jovian moon Europa have been used to refine models of Europa's interior. Europa is most likely differentiated into a metallic core surrounded by a rock mantle and a water ice-liquid outer shell, but the data cannot eliminate the possibility of a uniform mixture of dense silicate and metal beneath the water ice-liquid shell. The size of a metallic core is uncertain because of its unknown composition, but it could be as large as about 50 percent of Europa's radius. The thickness of Europa's outer shell of water ice-liquid must lie in the range of about 80 to 170 kilometers.     

Effects of injectio reduqing on plasma IL-8 and nitric oxide levels in rabbits with endotoxin induced disseminated intravascular coagulation

Experiments were performed for investigating the effects of Injectio Reduqing (RDQ) on plasma interleukin-8 (IL-8), NO2-/NO3-, complement 5a(C5a) and polymorphonuclear neutrophilic leukocyte (PMN) Chemotaxis Index (CI) in rabbits with endotoxin-induced disseminated intravascular coagulation (DIC). The results showed that plasma IL-8, NO2-/NO3-, C5a and CI levels of PMN increased markedly in model group, which were confirmed pathologically with obvious damage of tissues or organs. While in RDQ group the abov-mentioned parameters and damage of tissues or organs were reduced significantly (P < 0.01). The results suggested that the IL-8 and NO might be involved in pathogenesis of endotoxin-induced DIC, and RDQ could be used in preventing or treating DIC through mechanism of regulation of cytokines network.     

Emission characteristics of some dilute tungsten alloys

The effective work functions of nine tungsten-base alloys were measured. The second constituents were: 1) 5 pct Re, 2) 15 pct Re, 3) 2.5 pct Os, 4) 5 pct Os, 5) 1 pct Ir, 6) 2 pct Ir, 7) 5 pct Ta, 8) 10 pct Ta, and 9) 26 pct Ta. Work functions were determined from vacuum emission vehicle and thermionic emission microscope measurements. Mosaics of each surface were produced from the microscope which show the grain structure of the alloys and some anomalous emission areas. Some exceptionally high bare work functions were observed from samples with small alloy additions. For example, the work function of W1 pct Ir was 5.3 eV. DEAN LEO JACOBSON, formerly Section Manager, Electro-Optical Systems, Division of Xerox Corp., Pasadena, Calif.