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21.
Reference concentrations of cholecalciferol in animals: a basis for establishing non-target exposure
AAC Fairweather CT Eason PA Elder CMF Eason D Arthur 《New Zealand journal of zoology.》2013,40(4):280-289
AbstractCholecalciferol (vitamin D3) is widely used as a vertebrate pesticide in New Zealand. However, cholecalciferol also occurs naturally in animals. Therefore, when trying to determine whether a non-target animal has been exposed to cholecalciferol baits, knowledge of the baseline cholecalciferol concentrations in the animal's plasma and tissue is required. We analysed cattle, sheep, pig, deer, dog and cat plasma and liver samples for the vitamin D3 metabolite 25-hydroxycholecalciferol (25-OHD), a sensitive biomarker for cholecalciferol. Based on these data and a literature search we present 25-OHD reference ranges. We also examined the literature for 25-OHD concentrations in poisoned animals and compared these to the reference ranges. Where plasma and liver samples have 25-OHD concentrations at least four times higher than our reference ranges it is likely that the animal has been exposed to cholecalciferol baits. 25-OHD concentrations 10 times higher than the reference range indicate ingestion of abnormally high amounts of cholecalciferol. 相似文献
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Isolation of two populations of sperm cells and microelectrophoresis of pairs of sperm cells from pollen tubes of tobacco (Nicotiana tabacum) 总被引:1,自引:1,他引:0
Prior research has indicated that the two sperm cells of Nicotiana tabacum are dimorphic, suggesting that they may participate in preferential fertilization during in vivo fusion with the egg and
central cells. To probe the mechanism of potential preferential fertilization in this plant, it will be necessary to use modern
sensitive molecular techniques. For this purpose, two individual populations of two sperm cells, constituting the Svn (associated
with the vegetative nucleus) and Sua (unassociated with the vegetative nucleus), were isolated in the thousands from tobacco
pollen tubes with a micromanipulator as a preliminary step toward research on gametic recognition using molecular techniques.
Microelectrophoresis of paired sperm cells from a single pollen tube was conducted at different developmental stages. Sperm
cells isolated from 1-, 2-, 3- and 4-cm stylar lengths migrated to the negative pole, with the Sua displaying significantly
greater electrophoretic mobility than the Svn, reflecting a more positively charged cell surface on the Sua. The sperm cells
isolated from 1-cm style are very sensitive to electron potential in an electrophoretic field, presumably reflecting that
they are still in a young state. Differences in cell surface charge between the Sua and Svn may be related with cell fate
during fertilization.
Supported by National Natural Science Foundation of CHINA (30170060) 相似文献
24.
S Senthilvel B Jayashree V Mahalakshmi P Sathish Kumar S Nakka T Nepolean CT Hash 《BMC plant biology》2008,8(1):119
Background
Pearl millet [Pennisetum glaucum (L.) R. Br.] is a staple food and fodder crop of marginal agricultural lands of sub-Saharan Africa and the Indian subcontinent. It is also a summer forage crop in the southern USA, Australia and Latin America, and is the preferred mulch in Brazilian no-till soybean production systems. Use of molecular marker technology for pearl millet genetic improvement has been limited. Progress is hampered by insufficient numbers of PCR-compatible co-dominant markers that can be used readily in applied breeding programmes. Therefore, we sought to develop additional SSR markers for the pearl millet research community. 相似文献25.
Naguleswaran A Alaeddine F Guionaud C Vonlaufen N Sonda S Jenoe P Mevissen M Hemphill A 《International journal for parasitology》2005,35(13):1459-1472
We have previously shown that treatment of Neospora caninum tachyzoites with the aspartyl protease inhibitor pepstatin A reduces host cell invasion [Naguleswaran, A., Muller, N., Hemphill, A., 2003. Neospora caninum and Toxoplasma gondii: a novel adhesion/invasion assay reveals distinct differences in tachyzoite-host cell interactions. Exp. Parasitol. 104, 149-158]. Pepstatin A-affinity-chromatography led to the isolation of a major band of approximately 52 kDa which was identified as a homologue of a previously described Toxoplasma gondii putative protein disulfide isomerase (TgPDI) through tandem mass spectrometry. A BLAST search against N. caninum expressed sequence tags (ESTs) on the ApiDots server using TgPDI cDNA as query sequence revealed a 2251 bp PDI-like consensus (NcPDI), which shows 94% identity to the T. gondii homologue. In N. caninum tachyzoites, NcPDI was found mainly in the soluble hydrophilic fraction. Immunofluorescence showed that expression of NcPDI was dramatically down-regulated in the bradyzoite stage, and immunogold-EM on tachyzoites localised the protein to the cytoplasm, mostly in close vicinity to the nuclear membrane, to the micronemes, and to the parasite cell surface. However, NcPDI was absent in rhoptries and dense granules. Preincubation of tachyzoites with the sulfhydryl blocker 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB), p-chloromercuribenzoic acid (pCMBA), and with the PDI inhibitor bacitracin reduced adhesion of parasites to host cells. In addition, incubation of N. caninum tachyzoites with affinity-purified anti-NcPDI antibodies reduced host cell adhesion. PDIs catalyse the formation, reduction or isomerisation of disulfide bonds. Many major components of the adhesion and invasion machinery of apicomplexan parasites are cysteine-rich and dependent on correct folding via disulfide bond formation. Thus, our data points towards an important role for surface-associated NcPDI in Neospora-host cell interaction. 相似文献
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The IgG1hybridoma antibody, 91.9H, was originally raised against sulfated
mucins isolated from normal human colonic mucosa. Previous studies have
shown that the 91.9H antigen is expressed on normal colonic epithelial
cells and the sulfomucins that they produce, but not in the normal small
intestine and stomach. Tissue-specific changes occur in 91.9H antigen
expression in disease: the antigen diminishes in colonic carcinomas,
whereas in regions of gastric mucosa showing intestinal metaplasia and in
gastric carcinomas, the antigen is expressed as a "neo-antigen." This
report is concerned with elucidation, by the neoglycolipid technology, of
the determinant recognized by antibody 91.9H using sulfated and sialyl
oligosaccharides of Lewisa(Lea) and Lextypes, and analogs that lack
sulfate, sialic acid, or fucose. Binding experiments with the lipid-linked
oligosaccharides immobilized on chromatograms or on microwells, and
inhibition of binding experiments with free oligosaccharides based on di-,
tri- and tetrasaccharide backbones, show that the 91.9H antigenic
determinant is based on a trisaccharide backbone, and consists of the
3'-sulfated Leatetrasaccharide sequence, which is a potent ligand for the
E- and L-selectins. The antibody gives a relatively low signal with the
3'-sulfated non-fucosylated backbone, and has no detectable cross- reaction
with the 3'-sulfated Lexisomer, nor with sialyl-Leaand - Lexanalogues.
Antibody 91.9H is a valuable addition, therefore, to the repertoire of
reagents for mapping details of the distribution, and determining the
relative importance of sulfated and sialyl oligosaccharides as ligands for
the selectins, in normal and pathological epithelia and endothelia.
相似文献
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Tineke E Buffart Melanie Louw Nicole CT van Grieken Marianne Tijssen Beatriz Carvalho Bauke Ylstra Heike Grabsch Chris JJ Mulder Cornelis JH van de Velde Schalk W van der Merwe Gerrit A Meijer 《BMC medical genomics》2011,4(1):7
Background
Infection with H. pylori is important in the etiology of gastric cancer. Gastric cancer is infrequent in Africa, despite high frequencies of H. pylori infection, referred to as the African enigma. Variation in environmental and host factors influencing gastric cancer risk between different populations have been reported but little is known about the biological differences between gastric cancers from different geographic locations. We aim to study genomic instability patterns of gastric cancers obtained from patients from United Kingdom (UK) and South Africa (SA), in an attempt to support the African enigma hypothesis at the biological level.Methods
DNA was isolated from 67 gastric adenocarcinomas, 33 UK patients, 9 Caucasian SA patients and 25 native SA patients. Microsatellite instability and chromosomal instability were analyzed by PCR and microarray comparative genomic hybridization, respectively. Data was analyzed by supervised univariate and multivariate analyses as well as unsupervised hierarchical cluster analysis.Results
Tumors from Caucasian and native SA patients showed significantly more microsatellite instable tumors (p < 0.05). For the microsatellite stable tumors, geographical origin of the patients correlated with cluster membership, derived from unsupervised hierarchical cluster analysis (p = 0.001). Several chromosomal alterations showed significantly different frequencies in tumors from UK patients and native SA patients, but not between UK and Caucasian SA patients and between native and Caucasian SA patients.Conclusions
Gastric cancers from SA and UK patients show differences in genetic instability patterns, indicating possible different biological mechanisms in patients from different geographical origin. This is of future clinical relevance for stratification of gastric cancer therapy.30.
Mitochondrial DNA (mtDNA) sequences that include (a) a part of the
cytochrome b gene, (b) two tRNA genes, and (c) a part of the noncoding
D-loop region of 31 Anguilla japonica (Japanese eel) and 1 A. marmorata
collected from Taiwan, Japan, and mainland China were determined to
evaluate the population structure of Japanese eel. Among 30 genotypes
identified from the 31 Japanese eel mtDNAs sequenced, there are 58 variable
sites, predominantly clustered at the D-loop region. The phylogenetic tree
constructed by the unweighted pair-group method with arithmetic mean shows
neither significant genealogical branches nor geographic clusters.
Furthermore, the sequence-statistics test reveals little, if any,
significant genetic differentiation. These results indicate that the 31
Japanese eels might come from a single population. Analysis of sequence
variation in mtDNA by using the relationship between the number of
segregating sites and the average number of nucleotide differences under
the neutral mutation hypothesis reveals that neutral mutation acts as a
major factor influencing the evolutionary divergence of the Japanese eel
mitochondrial genome sequenced, especially in the noncoding region.
相似文献