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31.
Typical preparation of seed samples for infrared (IR) microspectroscopy involves imbibition of the seed for varying time periods followed by cryosectioning. Imbibition, however, may initiate germination even at 4° C with associated changes in the chemistry of the sample. We have found that it is possible to section seeds that are sufficiently hard, such as soybeans, on a standard laboratory microtome without imbibition. The use of dry sectioning of unimbibed seeds is reported here, as well as a comparison of different mounting media and modes of analysis. Glycerol, Tissue-Tek, and ethanol were used as mounting media, and the quality of the resulting spectra was assessed. Ethanol was the preferred mountant, because it dried quickly with no residue and thus did not interfere with the spectrum of interest. Analysis in transmission mode using barium fluoride windows to hold the samples was compared with transmission-reflection analysis with sections mounted on special infrared-reflecting slides. The two modes of analysis performed well in different regions of the spectrum. The mode of analysis (transmission vs. transmission-reflection) should be based on the components of greatest interest in the sample. 相似文献
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G van der Horst B Skosana A Legendre P Oyeyipo SS du Plessis 《Biotechnic & histochemistry》2018,93(1):49-58
We used automated sperm morphology analysis to investigate rat sperm morphometry and morphology in Sprague-Dawley and Wistar rats in three research centers to develop normal baseline values for sperm morphometry and to quantify the percentage of morphologically normal sperm in healthy rats. The participating centers were IRSN in Paris, France (Sprague-Dawley rats), University of the Western Cape, South Africa (Wistar rats) and Stellenbosch University (Wistar rats), South Africa. All three centers used identical sperm isolation techniques from the cauda epididymis, the same staining protocols, identical computer-aided sperm morphometry analysis (CASMA) software and microscopes with similar optics. With CASMA, fully automated analysis of the different parts of stained sperm, e.g., head, acrosome, mid-piece, can be performed, many sperm morphometric features can be measured accurately and eventually normal sperm morphology can be defined. We found that it is possible to distinguish sperm morphometric characteristics of Sprague-Dawley and Wistar rats. We also developed cut-off values for evaluating the percentage normal sperm in these two rat strains using the automatic analysis mode. Normal sperm morphology varied between 67 and 74% by contrast with previous findings of > 90%. 相似文献
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Fernanda Fraga Campos Policarpo A Sales Junior Alvaro José Romanha Márcio SS Araújo Ezequias P Siqueira Jarbas M Resende Tania MA Alves Olindo A Martins-Filho Vera Lúcia dos Santos Carlos A Rosa Carlos L Zani Betania Barros Cota 《Memórias do Instituto Oswaldo Cruz》2015,110(1):65-74
Aiming to identify new sources of bioactive secondary metabolites, we isolated 82
endophytic fungi from stems and barks of the native Brazilian tree Caesalpinia
echinata Lam. (Fabaceae). We tested their ethyl acetate extracts in several in vitro
assays. The organic extracts from three isolates showed antibacterial activity
against Staphylococcus aureus and Escherichia coli [minimal inhibitory concentration
(MIC) 32-64 μg/mL]. One isolate inhibited the growth of Salmonella typhimurium (MIC
64 μg/mL) and two isolates inhibited the growth of Klebsiella oxytoca (MIC 64 μg/mL),
Candida albicans and Candida tropicalis (MIC 64-128 μg/mL). Fourteen extracts at a
concentration of 20 μg/mL showed antitumour activities against human breast cancer
and human renal cancer cells, while two isolates showed anti-tumour activities
against human melanoma cancer cells. Six extracts were able to reduce the
proliferation of human peripheral blood mononuclear cells, indicating some degree of
selective toxicity. Four isolates were able to inhibit Leishmania (Leishmania)
amazonensis and one isolate inhibited Trypanosoma cruzi by at least 40% at 20 μg/mL.
The trypanocidal extract obtained from Fusarium sp. [] culture was subjected
to bioguided fractionation, which revealed beauvericin as the compound responsible
for the observed toxicity of Fusarium sp. to T. cruzi. This depsipeptide showed a
half maximal inhibitory concentration of 1.9 μg/mL (2.43 μM) in a T. cruzi cellular
culture assay. KF611679相似文献
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Abdullah Alghasham Abdulaziz AM Al Salloom Ahmed SS Alghamadi 《Nucleosides, nucleotides & nucleic acids》2015,34(1):56-68
Present study probes the role of peroxynitrite (ONOO-)-modified thymidine-5′-monophosphate (TMP) in SLE patients with different disease activity scores according to the SLE Disease Activity Index (SLEDAI). Serum analysis showed significant increased number of subjects positive for anti-ONOO--TMP-protein antibodies in SLE patients with different SLEDAI scores. Interestingly, the levels of these antibodies were significantly higher among SLE patients, whose SLEDAI scores were ≥20. In addition, a significant correlation was observed between the levels of anti-ONOO--TMP-protein antibodies and the SLEDAI score (r = 0.595, p < 0.0001). In short, this study shows a positive association between anti-ONOO--TMP-protein antibodies and SLEDAI. The stronger response observed in patients with higher SLEDAI scores suggests that anti-ONOO--TMP-protein antibodies may be useful in evaluating the progression of SLE and in elucidating the mechanisms of disease pathogenesis. 相似文献
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EG Ciolac SS Mantuani CM Neiva CEL Verardi DM Pess?a-Filho L Pimenta 《Biology of sport / Institute of Sport》2015,32(2):103-108
The aim of the present study was to analyse the usefulness of the 6-20 rating of perceived exertion (RPE) scale for prescribing and self-regulating high-intensity interval training (HIT) in young individuals. Eight healthy young subjects (age = 27.5±6.7 years) performed maximal graded exercise testing to determine their maximal and reserve heart rate (HR). Subjects then performed two HIT sessions (20 min on a treadmill) prescribed and regulated by their HR (HR: 1 min at 50% alternated with 1 min at 85% of reserve HR) or RPE (RPE: 1 minute at the 9-11 level [very light-fairly light] alternated with 1 minute at the 15-17 level [hard-very hard]) in random order. HR response and walking/running speed during the 20 min of exercise were compared between sessions. No significant difference between sessions was observed in HR during low- (HR: 135±15 bpm; RPE: 138±20 bpm) and high-intensity intervals (HR: 168±15 bpm; RPE: 170±18 bpm). Walking/running speed during low- (HR: 5.7±1.2 km · h−1; RPE: 5.7±1.3 km · h−1) and high-intensity intervals (HR: 7.8±1.9 km · h−1; RPE: 8.2±1.7 km · h−1) was also not different between sessions. No significant differences were observed in HR response and walking/running speed between HIT sessions prescribed and regulated by HR or RPE. This finding suggests that the 6-20 RPE scale may be a useful tool for prescribing and self-regulating HIT in young subjects. 相似文献
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Phagocytosis of bacteria by polymorphonuclear leukocytes: a freeze-fracture, scanning electron microscope, and thin-section investigation of membrane structure 总被引:2,自引:2,他引:0
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The changes in membrane structure of rabbit polymorphonuclear (PMN) leukocytes during bacterial phagocytosis was investigated with scanning electron microscope (SEM), thin-section, and freeze-fracture techniques. SEM observations of bacterial attachment sites showed the involvement of limited areas of PMN membrane surface (0.01-0.25μm(2)). Frequently, these areas of attachment were located on membrane extensions. The membrane extensions were present before, during, and after the engulfment of bacteria, but were diminished in size after bacterial engulfment. In general, the results obtained with SEM and thin-section techniques aided in the interpretation of the three-dimensional freeze-fracture replicas. Freeze-fracture results revealed the PMN leukocytes had two fracture faces as determined by the relative density of intramembranous particles (IMP). Membranous extensions of the plasma membrane, lysosomes, and phagocytic vacuoles contained IMP's with a distribution and density similar to those of the plasma membrane. During phagocytosis, IMPs within the plasma membrane did not undergo a massive aggregation. In fact, structural changes within the membranes were infrequent and localized to regions such as the attachment sites of bacteria, the fusion sites on the plasma membrane, and small scale changes in the phagocytic vacuole membrane during membrane fusion. During the formation of the phagocytic vacuole, the IMPs of the plasma membrane appeared to move in with the lipid bilayer while maintaining a distribution and density of IMPs similar to those of the plasma membranes. Occasionally, IMPs were aligned to linear arrays within phagocytic vacuole membranes. This alignment might be due to an interaction with linearly arranged motile structures on the side of the phagocytic vacuole membranes. IMP-free regions were observed after fusion of lysosomes with the phagocytic vacuoles or plasma membrane. These IMP-free areas probably represent sites where membrane fusion occurred between lysosomal membrane and phagocytic vacuole membrane or plasma membrane. Highly symmetrical patterns of IMPs were not observed during lysosomal membrane fusion. 相似文献
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The enzyme activity of Mg-ATPase, Na+ -K+ -ATPase, 5'-nucleotidase and NAD(P)H-oxidase was cytochemically detected at the ultrastructural level in mouse peritoneal macrophages infected with untreated and with specific antibody-coated Toxoplasma gondii tachyzoites. The Mg++ -ATPase and 5'-nucleotidase were distributed throughout the macrophages'plasma membrane but were not observed in the membrane lining endocytic vacuoles containing ingested parasites; however, Na+ -K+ -ATPase activity was detected in the macrophages'plasma membrane as well as in the parasitophorous vacuoles that contained untreated or specific antibody-coated parasites. Reaction product, indicative of NAD(P)H-oxidase, was detected in the parasitophorous vacuoles that contained only-specific antibody-coated parasites. 相似文献
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Veena?DhawanEmail author SS?Handu Chandra?Kanwal?Nain Nirmal?Kumar?Ganguly 《Molecular and cellular biochemistry》2005,269(1):1-11
Chronic exposure to l-arginine results in regression of atherosclerotic lesions and reversal of endothelial dysfunction. We investigated whether chronic l-arginine supplementation induces regression of atherosclerotic lesions and reversal of endothelial dysfunction in atherogenic rhesus monkeys and the mechanism which leads to these effects. About 12 male rhesus monkeys were fed 1% cholesterol and 18 g butter for 6 months to create an experimental model of hypercholesterolaemia and atherosclerosis (Group I) and 12 monkeys were fed standard stock diet for 6 months (Group II). After, 6 months these two groups were further divided into 2 sub-groups which in addition to their respective diets were fed 2.5% l-arginine in drinking water for additional 6 months (Group III and Group IV). Systemic nitric oxide (NO) formation was assessed as plasma nitrite and cGMP formation every 3 months. Oxygen free radical (OFR) generation and malondialdehyde production as an index of lipid peroxidation were determined. Changes in isometric tension were compared in isolated ring segments of thoracic aorta from normal and hypercholesterolemic animals.Cholesterol feeding progressively reduced plasma nitrite and cGMP generation (p<0.05). Dietary l-arginine partly restored the levels of plasma nitrite and cGMP (p<0.05) but did not change plasma cholesterol levels. l-arginine significantly reduced aortic intimal thickening, blocked the production of carotid and coronary intimal plaques and completely preserved endothelium-dependent vasodilator function. Further, l-arginine significantly inhibited generation of the reactive oxygen species (ROS) generation and lipid peroxidation.Chronic oral supplementation with l-arginine blocks the progression of plaques via restoration of nitric oxide synthase substrate availability and reduction of vascular oxidative stress. (Mol Cell Biochem 269: 1–11, 2005) 相似文献
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