Integration of the simian virus 40 genome into cellular DNA in temperature-sensitive (N) and temperature-insensitive (A) transformants of 3T3 rat and Chinese hamster lung cells

We studied the pattern of integration of the simian virus 40 (SV40) genome into the cellular DNA of N-transformants (temperature sensitive) and A-transformants (temperature insensitive) derived from 3T3-Fisher rat and Chinese hamster lung cells. The SV40 DNA was covalently linked to the cellular DNA in both types of transformants. In the rat cells, most N-transformants contained SV40 sequences integrated at a single site; most A-transformants contained SV40 sequences integrated at two to five sites. In the Chinese hamster cells, no significant correlation between the number of integration sites and the phenotype of the transformant was found; one of three integration sites were observed for both the N- and A-transformants. Single copies and tandem repeats of SV40 sequences were observed in A- and N-transformants derived from rat cells. A-transformants arise neither by amplification of the SV40 genome nor by integration at a unique site.     

Factors which disorganize microtubules or microfilaments increase the frequency of cell transformation by polyoma virus.

Griseofulvin, 12-O-tetradecanoyl phorbol-13-acetate, melittin, epidermal growth factor, vinblastine, cytochalasin B, podophyllotoxin, colcemid, and colchicine were unable to transform cells but could increase from 8- to 40-fold the frequency of cell transformation by polyoma virus. The 3T3-like cells were resting at confluence and were exposed to the drug only during the 1st week after viral infection. Griseofulvin, a tumor promoter, reduced or increased the frequency of transformation depending on the dose with which the infected cells were treated. The antitumor activity of tumor promoters is discussed.     

RNase P cleaves the adenine riboswitch and stabilizes pbuE mRNA in Bacillus subtilis

RNase P from Bacillus subtilis cleaves in vitro the adenine riboswitch upstream of pbuE, which codes for an adenine efflux pump. The guanine riboswitch, encoded upstream of xpt-pbuX operon, is not cleaved. The cleavage sites do not occur at any predicted structures that should be recognized by RNase P in the theoretical model of the adenine riboswitch. However, it is possible to draw alternative secondary structure models that match the apparent requirements for RNase P substrates at these cleavage sites. Support for these models is provided by appropriate mutagenesis experiments. Adenine showed no effect on the cleavage in vitro of the pbuE adenine riboswitch by RNase P holoenzyme from B. subtilis. The results of genetic experiments performed in B. subtilis support the cleavage of adenine riboswitch by RNase P in vivo and suggest that it induces the stabilization of pbuE mRNA under normal conditions.     

Schistosoma haematobium (Egyptian strain): rate of development and effect of praziquantel treatment

This study investigates the development of the Egyptian strain of Schistosoma haematobium and the resultant immunohistopathology and biochemical changes in organs affected. In addition, the response of different developmental stages of S. haematobium worms to praziquantel (PZQ) was examined. Schistosoma haematobium-infected hamsters were classified into 4 groups and were treated at day 35, 55, 75, and 95 postinfection (PI), respectively. Each group was subdivided into 3 subgroups. Two of them were treated orally with PZQ (300 mg/kg or 500 mg/kg divided equally on 2 consecutive days), and the third group was left without treatment. Treated groups were killed 20 days posttreatment. Infection with S. haematobium became patent 73 days PI; tissue egg load and worm fecundity were higher at 95 days and maximal 115 days PI, with an oogram pattern comparable to that in Schistosoma mansoni infection. In the liver, small cellular granulomas were observed 75 days PI, with preponderance of CD4+ T-cell phenotypes. In the urinary bladder, only submucosal focal Brunn's-nest formation and angiogenesis without typical granulomas were observed. Ninety-five and 115 days PI, confluent granulomata with multiple eggs in the center were observed in the liver and urinary bladder, with a preponderance of CD8+ positive T cells in the liver and hyperplasia of the urinary bladder epithelium with cystitis cystica and papillae formation. One hundred percent worm eradication was recorded with the higher dose of PZQ in animals treated 75 and 95 days PI. In conclusion, in spite of the long prepatent period of the Egyptian strain of S. haematobium, sensitivity to PZQ was recorded soon after infection. Granulomata were similar to those of S. mansoni in the livers and urinary bladders, but they were confluent with multiple eggs in the centers, hyperplasia of the urinary bladder urothelium with cystitis cystica, papillae, and Brunn's-nest formation predictive of malignant changes with no hepatocyte dysplasia.     

Late Cretaceous (Cenomanian to Campanian) paleoenvironmental history of the Eastern Canadian margin of the Western Interior Seaway: bonebeds and anoxic events

Upper Cretaceous strata in the Pasquia Hills of the northern Manitoba Escarpment, eastern Saskatchewan, Canada provide a detailed paleoenvironmental and sea-level record of the eastern margin of the Western Interior Seaway. Sediments deposited during the Cenomanian/Turonian Greenhorn marine cycle are dominantly black mudstones deposited in a stratified water column, with bottom-water anoxia recurrently reaching into the photic zone. A middle Cenomanian sea-level lowstand event followed by transgression left a series of bonebeds within the Belle Fourche Member of the Ashville Formation, indicating a sedimentary environment starved of coarse siliciclastics. Maximum sea level resulted in the formation of limestone beds within the Favel Formation, further favoured by reduced terrigenous sediment input compared to the western margin. Limestone sedimentation was followed by a phase of increased freshwater input under lower sea level conditions, and reducing zoo- and phytoplankton diversities. During final Greenhorn regression, eastern Saskatchewan probably turned into a restricted basin severely limiting marine circulation. Poor or absent benthic foraminiferal assemblages and biomarker analysis suggest prevailing watermass stratification throughout the Cenomanian/Turonian transgressive/regressive cycle. This was caused either by a freshwater lid, stratification of Boreal and Tethyan-derived watermasses, or both, to various intensities affected by changing sea level. Basin oxygenation during Niobrara time varies between localities along the eastern margin as documented by presence/absence of benthic and planktic foraminifera.     

Type I IFN as a natural adjuvant for a protective immune response: lessons from the influenza vaccine model

The identification of natural adjuvants capable of selectively promoting an efficient immune response against infectious agents would represent an important advance in immunology, with direct implications for vaccine development, whose progress is generally hampered by the difficulties in defining powerful synthetic adjuvants suitable for clinical use. Here, we demonstrate that endogenous type I IFN is necessary for the Th1 type of immune response induced by typical adjuvants in mice and that IFN itself is an unexpectedly powerful adjuvant when administered with the human influenza vaccine, for inducing IgG2a and IgA production and conferring protection from virus challenge. The finding that these cytokines, currently used in patients, are necessary for full expression of adjuvant activity and are sufficient for the generation of a protective immune response opens new perspectives in understanding the basis of immunity and in vaccine development.     

A comparison of symmetry and flatness measurements in small electron fields by different dosimeters in electron beam radiotherapy

BackgroundSymmetry and flatness are two quantities which should be evaluated in the commissioning and quality control of an electron beam in electron beam radiotherapy. The aim of this study is to compare symmetry and flatness obtained using three different dosimeters for various small and large fields in electron beam radiotherapy with linac.Materials and methodsBeam profile measurements were performed in a PTW water phantom for 10, 15 and 18 MeV electron beams of an Elekta Precise linac for small and large beams (1.5 × 1.5 cm² to 20 × 20 cm² field sizes). A Diode E detector and Semiflex-3D and Advanced Markus ionization chambers were used for dosimetry.ResultsBased on the obtained results, there are minor differences between the responses from different dosimeters (Diode E detector and Semiflex-3D and Advanced Markus ionization chambers) in measurement of symmetry and flatness for the electron beams. The symmetry and flatness values increase with increasing field size and electron beam energy for small and large field sizes, while the increases are minor in some cases.ConclusionsThe results indicate that the differences between the symmetry and flatness values obtained from the three dosimeter types are not practically important.     

Hybrid E. coli--Mitochondrial ribonuclease P RNAs are catalytically active

RNase P is a ribonucleoprotein that cleaves tRNA precursors at their 5'-end. Mitochondrion-encoded RNA subunits of mitochondrial RNase P (mtP-RNA) have been identified in jakobid flagellates such as Reclinomonas americana, in the prasinophyte alga Nephroselmis olivacea, and in several ascomycete and zygomycete fungi. While the structures of ascomycete mtP-RNAs are highly reduced, those of jakobids, prasinophytes, and zygomycetes retain most conserved features of their bacterial counterparts. Therefore, these mtP-RNAs might be active in vitro in the absence of a protein subunit, as are bacterial P-RNAs. Here we present a comparative structural analysis including seven newly characterized jakobid mtP-RNAs. We investigate ribozyme activities of mtP-RNAs and find that even the most bacteria-like molecules of jakobids are inactive in vitro. However, when certain domains of jakobid and N. olivacea mtP-RNAs are replaced with those from Escherichia coli, these hybrid RNAs show catalytic activity. In vitro mutagenesis of these hybrid mtP-RNAs shows that various structural elements play a critical role in ribozyme catalysis and provide further support for the presence of these elements in mtP-RNAs. These include GNRA tetraloops in helix P14 and P18 of Jakoba libera, and a remnant P3 pairing in Seculamonas ecuadoriensis. Finally, we will discuss reasons for the failure of mtP-RNAs to show catalytic activity in the absence of P-proteins based on our mutagenesis analysis.