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141.
Nature of polymorphisms in 16S-23S rRNA gene intergenic transcribed spacer fingerprinting of Bacillus and related genera 总被引:1,自引:0,他引:1
Daffonchio D Cherif A Brusetti L Rizzi A Mora D Boudabous A Borin S 《Applied and environmental microbiology》2003,69(9):5128-5137
The intergenic transcribed spacers (ITS) between the 16S and 23S rRNA genetic loci are frequently used in PCR fingerprinting to discriminate bacterial strains at the species and intraspecies levels. We investigated the molecular nature of polymorphisms in ITS-PCR fingerprinting of low-G+C-content spore-forming bacteria belonging to the genera Bacillus, Brevibacillus, Geobacillus, and Paenibacillus: We found that besides the polymorphisms in the homoduplex fragments amplified by PCR, heteroduplex products formed during PCR between amplicons from different ribosomal operons, with or without tRNA genes in the ITS, contribute to the interstrain variability in ITS-PCR fingerprinting patterns obtained in polyacrylamide-based gel matrices. The heteroduplex nature of the discriminating bands was demonstrated by fragment separation in denaturing polyacrylamide gels, by capillary electrophoresis, and by cloning, sequencing, and recombination of purified short and tRNA gene-containing long ITS. We also found that heteroduplex product formation is enhanced by increasing the number of PCR cycles. Homoduplex-heteroduplex polymorphisms (HHP) in a conserved region, such as the 16S and 23S rRNA gene ITS, allowed discrimination of closely related strains and species undistinguishable by other methods, indicating that ITS-HHP analysis is an easy and reproducible additional tool for strain typing. 相似文献
142.
Bradbury A Velappan N Verzillo V Ovecka M Chasteen L Sblattero D Marzari R Lou J Siegel R Pavlik P 《Trends in biotechnology》2003,21(7):312-317
There are many ways in which the use of antibodies and antibody selection can be improved and developed for high-throughput characterization. Standard protocols, such as immunoprecipitation, western blotting and immunofluorescence, can be used with antibody fragments generated by display technologies. Together with novel approaches, such as antibody chips and intracellular immunization, these methods will yield useful proteomic data following adaptation of the protocols for increased reliability and robustness. To date, most work has focused on the use of standard, well-characterized commercial antibodies. Such protocols need to be adapted for broader use, for example, with antibody fragments or other binders generated by display technologies, because it is unlikely that traditional approaches will provide the required throughput. 相似文献
143.
Background
A large body of data derived from animal, epidemiological and clinical studies indicate that n-3 polyunsaturated fatty acids have a favourable effect on the prognosis of patients with cardiovascular disease in general, and on reducing sudden death in particular. Depressed heart rate variability (HRV), an indicator of impairment of the autonomic nervous system, has been shown to be a powerful predictor of subsequent mortality in patients surviving an acute myocardial infarction. A multitude of studies have demonstrated this strong association, suggesting that the imbalance in the sympathic/parasympathetic system may facilitate emergence of ventricular arrhythmias. Heart rate variability parameters will be assessed in the present study, with the primary objective of evaluating the possible superiority of Omacor (a highly refined, concentrated omega-3 fatty acid) versus placebo in improving HRV from baseline to endpoint in patients with recent uncomplicated myocardial infarction. Both groups will receive optimal conventional treatment. The study will also explore and quantify improvement in time domain HRV indices and will assess the safety of administering Omacor to optimally treated post-infarction patients (conventional treatment).Methods
This multi-centre study will evaluate the effect of Omacor 1 g, o.d. on time-domain HRV parameters in comparison to placebo o.d. in patients with recent uncomplicated transmural myocardial infarction. Patients will be screened during the first few days after the acute event as appropriate for the patient's condition, and after obtaining informed consent. Based on inclusion/exclusion criteria, a first 24-hour Holter recording will be performed. Two to five days later, screened patients still eligible for the study will undergo a second 24-hour Holter recording. After the second Holter recording, all patients will be randomly allocated to treatment with Omacor 1 g, o.d. or placebo o.d. One hundred patients will be followed in double-blind fashion for a six-month period after randomization. Visits, including 24-hour Holter recording and assessment of adverse events, will take place at one-month intervals ± five days after randomization, i.e., six times in all. 相似文献144.
Daniele?ArobbaEmail author Rosanna?Caramiello Angiolo?Del Lucchese 《Vegetation History and Archaeobotany》2003,12(4):253-262
The archaeological site we studied is part of an early Iron Age hill fort (8th/7th cent. b.c.), located 800 m from the coast on the top of a hill named MonteTrabocchetto. This paper concerns an excavation, called saggio O, which disclosed a very varied stratigraphy characterised by highly anthropogenic layers and by a pit, presumably used as a silo for food storage, which was very rich in charred seeds and fruits. The study of the pit content showed the dominance of Hordeum vulgare, while Triticum dicoccon, T. monococcum, T. aestivum/durum, Panicum miliaceum and Setaria italica were less strongly represented. Some edible Leguminosae were also found (Lens culinaris, Vicia faba var. minor and V. ervilia). In the frequented areas around the pit, herbaceous weeds and fruit tree macro-remains were present (Prunus cf. spinosa, Corylus avellana, Quercus sp. and Vitis vinifera ssp. sylvestris). The identification of a large number of botanical taxa has provided important information on food of plant origin and agricultural practices during the early Iron Age on the Ligurian coast, the proto-historic archaeobotanical aspects of which are largely unknown. 相似文献
145.
Di Fabio R Tranquillini E Bertani B Alvaro G Micheli F Sabbatini F Pizzi MD Pentassuglia G Pasquarello A Messeri T Donati D Ratti E Arban R Dal Forno G Reggiani A Barnaby RJ 《Bioorganic & medicinal chemistry letters》2003,13(21):3863-3866
To identify neuroprotective agents after stroke, new substituted tetrahydroquinoline derivatives were designed as antagonists of the glycine binding site associated to the NMDA receptor, satisfying the key pharmacophoric requirements. In particular, the racemate 3c exhibited outstanding in vivo activity in the MCAo model in rats, when given iv both pre- and post-ischemia. Pure enantiomers 3c-(+) and 3c-(-) have been prepared following an original synthetic route. Despite the significant difference of activity observed in vitro, they shown similar neuroprotective profile in the MCAo model in rats. 相似文献
146.
Semen from 5 Piedmontese bulls was subjected to the hypoosmotic swelling (HOS) test in order to determine if the results could be correlated to the fertilizing capacity in vitro. Semen was routinely prepared for in vitro fertilization (IVF), with aliquots being sampled after thawing, after separation on a Percoll gradient and after capacitation in a medium containing heparin. The aliquots were added to a fructose-sodium citrate hypoosmotic solution (100 mOsm) and incubated at 37 degrees C for 5 min. At least 200 spermatozoa were observed at x 400 and classified according to the presence or the absence of a swollen tail. After capacitation, spermatozoa were used to fertilize in vitro-matured bovine oocytes (1.5 x10(6) cells/mL); IVF was performed in Fert-TALP medium supplemented with 6 mg/mL BSA and 10 microL/mL heparin in a humidified atmosphere with 5% CO2 and 5% oxygen. Presumptive zygotes were cultured in SOF medium supplemented with 8 mg/mL BSA and amino acids. There were no significant differences in the in vitro fertility of the bulls, but a significant difference was found between bulls in the response to the HOS test. The 3 assays were significantly correlated, while no significant correlation was observed between the percentage of swollen spermatozoa and in vitro fertility. The HOS test does not appear to be sufficiently sensitive to discriminate between semen samples of intermediate fertility like those used in this preliminary research. 相似文献
147.
The measurement of the Reynolds stress tensor, or at least of some of its components, is a necessary step to assess if the turbulence associated with the flow near prosthetic devices can damage blood constituents. Because of the intrinsic three dimensionality of turbulence, in general, a three-component anemometer should be used to measure directly the components of the Reynolds stress tensor. However, this can be practically unfeasible, especially in vivo; therefore, it is interesting to investigate the possibility of characterizing the turbulent flows that may occur in the circulatory system with the monodimensional data that a less complete equipment (e.g., a pulsed ultrasound Doppler) can yield. From the general expression of the Reynolds stress tensor, the highest shear stress can be deduced, as well as the Reynolds normal stress in the main flow direction. The relation between these two quantities, which is an issue already addressed in previous works, can thus be rigorously formulated in terms of some characteristic parameters of the Reynolds stress tensor, the principal normal stresses and the angles that the directions that define them form with the main flow direction. An experimental verification of the ratio of the two above-mentioned quantitites for the flow across bileaflet valves, investigated by means of two-dimensional laser Doppler anemometry, will illustrate the limitations of the monodimensional approach estimating the maximum load on blood constituents. 相似文献
148.
Daniele Castagnolo Fabrizio Manetti Marco Radi Beatrice Bechi Mafalda Pagano Alessandro De Logu Rita Meleddu Manuela Saddi Maurizio Botta 《Bioorganic & medicinal chemistry》2009,17(15):5716-5721
Two series of novel rigid pyrazolone derivatives were synthesized and evaluated as inhibitors of Mycobacterium tuberculosis (MTB), the causative agent of tuberculosis. Two of these compounds showed a high activity against MTB (MIC = 4 μg/mL). The newly synthesized pyrazolones were also computationally investigated to analyze if their properties fit the pharmacophoric model for antitubercular compounds previously built by us. The results are in agreement with those reported by us previously for a class of pyrazole analogues and confirm the fundamental role of the p-chlorophenyl moiety at C4 in the antimycobacterial activity. 相似文献
149.
Daniele Dondi Daniele Merli Luca Pretali Maurizio Fagnoni Angelo Albini Nick Serpone 《Photochemical & photobiological sciences》2007,6(11):1210-1217
A series of prebiotic mixtures of simple molecules, sources of C, H, N, and O, were examined under conditions that may have prevailed during the Hadean eon (4.6-3.8 billion years), namely an oxygen-free atmosphere and a significant UV radiation flux over a large wavelength range due to the absence of an ozone layer. Mixtures contained a C source (methanol, acetone or other ketones), a N source (ammonia or methylamine), and an O source (water) at various molar ratios of C : H : N : O. When subjected to UV light or heated for periods of 7 to 45 days under an argon atmosphere, they yielded a narrow product distribution of a few principal compounds. Different initial conditions produced different distributions. The nature of the products was ascertained by gas chromatographic-mass spectral analysis (GC-MS). UVC irradiation of an aqueous methanol-ammonia-water prebiotic mixture for 14 days under low UV dose (6 x 10(-2) Einstein) produced methylisourea, hexamethylenetetramine (HMT), methyl-HMT and hydroxy-HMT, whereas under high UV dose (45 days; 1.9 x 10(-1) Einstein) yielded only HMT. By contrast, the prebiotic mixture composed of acetone-ammonia-water produced five principal species with acetamide as the major component; thermally the same mixture produced a different product distribution of four principal species. UVC irradiation of the CH(3)CN-NH(3)-H(2)O prebiotic mixture for 7 days gave mostly trimethyl-s-triazine, whereas in the presence of two metal oxides (TiO(2) or Fe(2)O(3)) also produced some HMT; the thermal process yielded only acetamide. 相似文献
150.
Daniele Moretti Barbara Del Bello Giulia Allavena Alessandro Corti Cinzia Signorini Emilia Maellaro 《PloS one》2015,10(2)
Calpain-3 is an intracellular cysteine protease, belonging to Calpain superfamily and predominantly expressed in skeletal muscle. In human melanoma cell lines and biopsies, we previously identified two novel splicing variants (hMp78 and hMp84) of Calpain-3 gene (CAPN3), which have a significant lower expression in vertical growth phase melanomas and, even lower, in metastases, compared to benign nevi. In the present study, in order to investigate the pathophysiological role played by the longer Calpain-3 variant, hMp84, in melanoma cells, we over-expressed it in A375 and HT-144 cells. In A375 cells, the enforced expression of hMp84 induces p53 stabilization, and modulates the expression of a few p53- and oxidative stress-related genes. Consistently, hMp84 increases the intracellular production of ROS (Reactive Oxygen Species), which lead to oxidative modification of phospholipids (formation of F2-isoprostanes) and DNA damage. Such events culminate in an adverse cell fate, as indicated by the decrease of cell proliferation and by cell death. To a different extent, either the antioxidant N-acetyl-cysteine or the p53 inhibitor, Pifithrin-α, recover cell viability and decrease ROS formation. Similarly to A375 cells, hMp84 over-expression causes inhibition of cell proliferation, cell death, and increase of both ROS levels and F2-isoprostanes also in HT-144 cells. However, in these cells no p53 accumulation occurs. In both cell lines, no significant change of cell proliferation and cell damage is observed in cells over-expressing the mutant hMp84C42S devoid of its enzymatic activity, suggesting that the catalytic activity of hMp84 is required for its detrimental effects. Since a more aggressive phenotype is expected to benefit from down-regulation of mechanisms impairing cell growth and survival, we envisage that Calpain-3 down-regulation can be regarded as a novel mechanism contributing to melanoma progression. 相似文献