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61.
In Caenorhabditis elegans, an effective RNA interference (RNAi) response requires the production of secondary short interfering RNAs (siRNAs) by RNA-directed RNA polymerases (RdRPs). We cloned secondary siRNAs from transgenic C. elegans lines expressing a single 22-nucleotide primary siRNA. Several secondary siRNAs start a few nucleotides downstream of the primary siRNA, indicating that non-RISC (RNA-induced silencing complex)-cleaved mRNAs are substrates for secondary siRNA production. In lines expressing primary siRNAs with single-nucleotide mismatches, secondary siRNAs do not carry the mismatch but contain the nucleotide complementary to the mRNA. We infer that RdRPs perform unprimed RNA synthesis. Secondary siRNAs are only of antisense polarity, carry 5' di- or triphosphates, and are only in the minority associated with RDE-1, the RNAi-specific Argonaute protein. Therefore, secondary siRNAs represent a distinct class of small RNAs. Their biogenesis depends on RdRPs, and we propose that each secondary siRNA is an individual RdRP product.  相似文献   
62.
Caenorhabditis elegans proteins AFF-1 and EFF-1 [C. elegans fusion family (CeFF) proteins] are essential for developmental cell-to-cell fusion and can merge insect cells. To study the structure and function of AFF-1, we constructed vesicular stomatitis virus (VSV) displaying AFF-1 on the viral envelope, substituting the native fusogen VSV glycoprotein. Electron microscopy and tomography revealed that AFF-1 formed distinct supercomplexes resembling pentameric and hexameric "flowers" on pseudoviruses. Viruses carrying AFF-1 infected mammalian cells only when CeFFs were on the target cell surface. Furthermore, we identified fusion family (FF) proteins within and beyond nematodes, and divergent members from the human parasitic nematode Trichinella spiralis and the chordate Branchiostoma floridae could also fuse mammalian cells. Thus, FF proteins are part of an ancient family of cellular fusogens that can promote fusion when expressed on a viral particle.  相似文献   
63.
A specific immunofluorescent histochemical method for cyclic adenosine monophosphate was used to study rat cerebellum. After topical treatment with norepinephrine or stimulation of norepinephrine-containing afferents from locus coeruleus, there was a striking increase in the number of Purkinje cells with strong cyclic adenosine monophosphate reactivity. Other putative inhibitory transmitters had no significant effect on staining of Purkinje cells. The results provide the first histochemical support for the hypothesis that cyclic adenosine monophosphate can be generated postsynaptically in central neurons in response to noradrenergic stimuli.  相似文献   
64.
To assess the antinociceptive efficacy of transdermal (TD) buprenorphine (B) in dogs, a prospective, positive-controlled experimental study was performed in 10 healthy Beagles. In an open label crossover design, the dogs initially received intravenous B (IVB, 0.02 mg kg?1) as a positive control, followed by TDB (52.5 μg h?1) 4 months later. Blood was collected at regular intervals for determination of the plasma concentrations of B ([B]) and its metabolite norbuprenorphine. The antinociceptive efficacy was assessed using thermal and mechanical models of nociception. The peak concentration [B] was 1.54 ng mL?1 (±1.98) 60 h after TDB application, although three dogs had no measurable [B] after TDB. Maximum thermal threshold (TT) was 52.6 °C (±0.48) at 1 h after IVB administration and 51.63 °C (±1.01) 72 h after TDB application. The significant increase in TT indicated that effective antinociception was achieved beyond 36 h after the application of TDB, lasting until patch removal. There was hysteresis between [B] and the antinociceptive effect.  相似文献   
65.
Wet deposition plays an important role as a mechanism for cleansing of the polluted troposphere. Fluxes of pollutants from the atmosphere to earth's surface can affect soil and aquatic ecosystems. The investigation of the pollutant incorporation into wet deposition is therefore of considerable interest for environmental research. The project “Winter precipitation at Mount Rigi” addresses the processes determining the chemical composition of wet deposition through an integrated approach considering atmospheric physical and chemical processes and their complex interactions. The investigated topics include: aerosol and gas scavenging, atmospheric chemistry (cloud water chemistry), cloud physics and dynamics. The design of the experiment, using three ground based measuring stations at the northwestern slope of the mountain, and the conceptual ideas using case study investigations are discussed.  相似文献   
66.
The concentrations of major ions and spheroidal carbonaceous fly-ash particles (SCPs) in bulk deposition were determined in weekly samples from six European mountain lakes during 1997/98. SCPs are produced only from high temperature combustion of fossil-fuels and therefore provide an unambiguous indicator of atmospheric deposition from this source. Positive correlations were observed between SCPs and SO4 2?, NO3 ? and NH4 + at all sites except for some determinands at Jorisee (Switzerland) and Starolesnienske (Slovakia). Correlations between SCPs and SO4 2? + NO3 ? were always more positive than for SCPs with 'total acid ions' (SO4 2? + NO3 ? + NH4 +). This is in agreement with the expectation that the contribution to NH4 + deposition made by fossil-fuels is negligible. Good positive correlations between SCPs and all acid anions were observed at Estany Redo (Pyrenees); lower but still positive correlations were observed for all acid ions with SCPs at Gossenköllesee (Austria), Lochnagar and Kårvatn (central Norway), whilst little trend in correlation was observed for Jorisee and Starolesnienske. It is suggested that this gradient reflects the influence of fossil-fuels on acid deposition in these areas. A high positive correlation was observed between SCP and Cl? at Gossenkollesee possibly as a result of HCl from coal combustion.  相似文献   
67.
Cold-pressed, non-raffinated evening primrose oil was found to contain lipophilic radical scavengers. A highly enriched fraction of these compounds could be obtained from the oil by extraction with aqueous ethanol and subsequent liquid-liquid partitioning with petroleum. LC-DAD-MS analysis revealed that the fraction contained three aromatic compounds with identical UV and ESI-MS spectra. The compounds were isolated by RP-HPLC and their structures established by chemical and spectroscopic means as 3-O-trans-caffeoyl derivatives of betulinic, morolic, and oleanolic acid. The morolic acid derivative was a new compound. The three esters exhibited pronounced radical scavenging activity against the stable 2,2-diphenyl-1-picrylhydrazyl radical and were potent inhibitors of neutrophil elastase and cyclooxygenase-1 and -2 in vitro. Commercial samples of evening primrose oils contained only traces of these lipophilic antioxidants.  相似文献   
68.
Aerosol photoemission (APE) has been found to be an excellent surrogate measure of particulate PAH concentration in combustion aerosols under a variety of laboratory and field conditions. Samples of oil stove, automobile exhaust, and urban air particulate matter were concurrently analyzed for aerosol photoemission intensity and particulate PAH concentration. In-situ, real-time analysis of the particulate matter was performed by UV-induced electron photoemission. Two photon energies, 4.9 and 6.7 eV, were chosen for irradiation. The latter energy was more sensitive and less selective. Particulate PAH concentrations were determined by collecting the particulate matter on a filter, solvent extracting the filter, and analyses of the extracts by gas chromatography / mass spectroscopy. Linear correlations (0.84≤r2≤1.00) were observed between aerosol photoemission intensities and the sum of particulate PAH concentrations for laboratory and field studies under a wide variety of conditions. The studies performed to date cover a dynamic range of three orders of magnitude (1 to 1000 ng / m3). The time resolution is in the order of a few seconds.  相似文献   
69.
Defence reactions of cattle against Fasciola hepatica take place not only in the liver but also on a prehepatic level. The aim of the present experiments is to identify and quantify the cellular response in the small intestine. Five groups of two bulls (Simmental x Red Holstein, 6 months old) were infected by oesophageal probe with metacercariae of F. hepatica (MC) according to two experimental procedures. Two groups of bulls served as controls. In the first experiment both infected groups received 1000 MC once or twice at an interval of five weeks. The intestinal tissue was taken before infection and 5, 10 and 19 weeks p.i., respectively 5 and 14 weeks p. reinf. (p.ri.). In the second experiment, three infected groups received 400, 2 x 400 MC (at an interval of five weeks) and 800 MC respectively. Biopsies were taken one week ante infectionem, 6, 11 and 19 weeks p.i., respectively 1, 6 and 14 weeks post reinfectionem (p. ri.). The biopsies of the small intestine were prepared for histology and the tissue sections stained as follows: Astra blue for mucosal mast cells (MMC), Lendrum for eosinophils (EOS) and Periodic Acid Schiff (PAS) for goblet cells (GC). The density of the three cell populations in the mucosa was determined microscopically and quantified. In both experiments the density of the MMC and EOS increased with each reinfection and depended on the infectious dose. There was no significant variation of the GC-numbers. The described intestinal cellular reactions suggest a participation of the small intestinal wall in the immunity against the common liver fluke.  相似文献   
70.
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