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61.
Aims: To develop a SYBR Green quantitative real-time PCR protocol enabling detection and quantification of a fish probiotic and two turbot pathogenic Vibrio spp. in microcosms.
Methods and Results: Phaeobacter 27-4, Vibrio anguillarum 90-11-287 and Vibrio splendidus DMC-1 were quantified as pure and mixed cultures and in presence of microalgae ( Isochrysis galbana ), rotifers ( Brachionus plicatilis ), Artemia nauplii or turbot ( Psetta maxima ) larvae by real-time PCR based on primers directed at genetic loci coding for antagonistic and virulence-related functions respectively. The optimized protocol was used to study bioencapsulation and maintenance of the probiont and pathogens in rotifers and for the detection and quantification of Phaeobacter and V. anguillarum in turbot larvae fed rotifers loaded with the different bacteria in a challenge trial.
Conclusions: Our real-time PCR protocol is reproducible and specific. The method requires separate standard curve for each host organism and can be used to detect and quantify probiotic Phaeobacter and pathogenic Vibrio bioencapsulated in rotifers and in turbot larvae.
Significance and Impact of the Study: Our method allows monitoring and quantification of a turbot larvae probiotic bacteria and turbot pathogenic vibrios in in vivo trials and will be useful tools for detecting the bacteria in industrial rearing units. 相似文献
Methods and Results: Phaeobacter 27-4, Vibrio anguillarum 90-11-287 and Vibrio splendidus DMC-1 were quantified as pure and mixed cultures and in presence of microalgae ( Isochrysis galbana ), rotifers ( Brachionus plicatilis ), Artemia nauplii or turbot ( Psetta maxima ) larvae by real-time PCR based on primers directed at genetic loci coding for antagonistic and virulence-related functions respectively. The optimized protocol was used to study bioencapsulation and maintenance of the probiont and pathogens in rotifers and for the detection and quantification of Phaeobacter and V. anguillarum in turbot larvae fed rotifers loaded with the different bacteria in a challenge trial.
Conclusions: Our real-time PCR protocol is reproducible and specific. The method requires separate standard curve for each host organism and can be used to detect and quantify probiotic Phaeobacter and pathogenic Vibrio bioencapsulated in rotifers and in turbot larvae.
Significance and Impact of the Study: Our method allows monitoring and quantification of a turbot larvae probiotic bacteria and turbot pathogenic vibrios in in vivo trials and will be useful tools for detecting the bacteria in industrial rearing units. 相似文献
62.
Jens T. Høeg Yair Achituv Benny K.K. Chan Karen Chan Peter Gram Jensen Marcos Pérez‐Losada 《Journal of morphology》2009,270(2):241-255
We used scanning electron microscopy (SEM) to describe cypris morphology in species of the barnacles Ibla and Paralepas, both of which are pivotal in understanding cirripede evolution. In Ibla, we also studied late naupliar stages with video and SEM. Special emphasis was put on the lattice organs, the antennules and the thorax and telson. In Paralepas we had settled specimens only and could therefore only investigate the carapace with the lattice organs. Cyprids of Ibla quadrivalvis and Paralepas dannevigi have five sets of lattice organs, grouped as two anterior and three posterior pairs. The organs are of the pore‐field type and the terminal pore is situated anteriorly in the first pair, just as in the Rhizocephala and the Thoracica. In Ibla the armament of antennular sensilla resembles that found in the Thoracica but differs from the Rhizocephala. The absence of setules on the A and B setae sited terminally on the fourth antennular segment is a similarity with the Acrothoracica. The attachment disc is angled rather than facing distally and is encircled by a low cuticular velum. The thoracopods have two‐segmented endopods and exopods as in the Thoracica, but the number, shape, and position of thoracopodal setae differ somewhat from other species of that superorder. Both Ibla and Paralepas cyprids have a deeply cleaved telson, but no independent abdominal part. In cypris morphology, Ibla and Paralepas show several synapomorphies with the clade comprising Rhizocephala and Thoracica and there are no specific apomorphies with either the Acrothoracica, the Rhizocephala or any particular subgroup within the Thoracica. This is in agreement with recent molecular evidence that Ibla (Ibliformes) is the sister taxon to all other Thoracica and the ibliforms therefore become the outgroup of choice for studying character evolution within the superorder. Paralepas, and other pedunculated barnacles without shell plates, are apparently not primitive but are secondarily evolved and nested within the Thoracica. J. Morphol., 2009. © 2008 Wiley‐Liss, Inc. 相似文献
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64.
Karin A. W. Wadt Lauren G. Aoude Lotte Krogh Lone Sunde Anders Bojesen Karen Gr?nskov Nine Wartacz Jakob Ek Morten Tolstrup-Andersen Mette Klarskov-Andersen ?ke Borg Steffen Heegaard Jens F. Kiilgaard Thomas V. O. Hansen Kerenaftali Klein G?ran J?nsson Krzysztof T. Drzewiecki Morten Dun? Nicholas K. Hayward Anne-Marie Gerdes 《PloS one》2015,10(3)
Both environmental and host factors influence risk of cutaneous
melanoma (CM), and worldwide, the incidence varies depending on constitutional determinants of skin type and pigmentation, latitude, and patterns of sun exposure. We performed genetic analysis of CDKN2A, CDK4, BAP1, MC1R, and MITFp.E318K in Danish high-risk melanoma cases and found CDKN2A germline mutations in 11.3% of CM families with three or more affected individuals, including four previously undescribed mutations. Rare mutations were also seen in CDK4 and BAP1, while MC1R variants were common, occurring at more than twice the frequency compared to Danish controls. The MITF p.E318K variant similarly occurred at an approximately three-fold higher frequency in melanoma cases than controls. To conclude, we propose that mutation screening of CDKN2A and CDK4 in Denmark should predominantly be performed in families with at least 3 cases of CM. In addition, we recommend that testing of BAP1 should not be conducted routinely in CM families but should be reserved for families with CM and uveal melanoma, or mesothelioma. 相似文献
65.
Henrik Lütken James R. Lloyd Mikkel A. Glaring Lone Baunsgaard Kristian Holst Laursen Anna Haldrup Jens Kossmann Andreas Blennow 《Planta》2010,232(5):1127-1139
Two glucanotransferases, disproportionating enzyme 1 (StDPE1) and disproportionating enzyme 2 (StDPE2), were repressed using
RNA interference technology in potato, leading to plants repressed in either isoform individually, or both simultaneously.
This is the first detailed report of their combined repression. Plants lacking StDPE1 accumulated slightly more starch in
their leaves than control plants and high levels of maltotriose, while those lacking StDPE2 contained maltose and large amounts
of starch. Plants repressed in both isoforms accumulated similar amounts of starch to those lacking StDPE2. In addition, they
contained a range of malto-oligosaccharides from maltose to maltoheptaose. Plants repressed in both isoforms had chlorotic
leaves and did not grow as well as either the controls or lines where only one of the isoforms was repressed. Examination
of photosynthetic parameters suggested that this was most likely due to a decrease in carbon assimilation. The subcellular
localisation of StDPE2 was re-addressed in parallel with DPE2 from Arabidopsis thaliana by transient expression of yellow fluorescent protein fusions in tobacco. No translocation to the chloroplasts was observed
for any of the fusion proteins, supporting a cytosolic role of the StDPE2 enzyme in leaf starch metabolism, as has been observed
for Arabidopsis DPE2. It is concluded that StDPE1 and StDPE2 have individual essential roles in starch metabolism in potato and consequently
repression of these disables regulation of leaf malto-oligosaccharides, starch content and photosynthetic activity and thereby
plant growth possibly by a negative feedback mechanism. 相似文献
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69.
?sa N??v Lena Erlandsson Josefin Axelsson Irene Larsson Martin Johansson Lena Wester-Rosenl?f Matthias M?rgelin Vera Casslén Magnus Gram Bo ?kerstr?m Stefan R. Hansson 《PloS one》2015,10(5)
Preeclampsia is one of the most serious pregnancy-related diseases and clinically manifests as hypertension and proteinuria after 20 gestational weeks. The worldwide prevalence is 3-8% of pregnancies, making it the most common cause of maternal and fetal morbidity and mortality. Preeclampsia lacks an effective therapy, and the only “cure” is delivery. We have previously shown that increased synthesis and accumulation of cell-free fetal hemoglobin (HbF) in the placenta is important in the pathophysiology of preeclampsia. Extracellular hemoglobin (Hb) and its metabolites induce oxidative stress, which may lead to acute renal failure and vascular dysfunction seen in preeclampsia. The human endogenous protein, α1-microglobulin (A1M), removes cell-free heme-groups and induces natural tissue repair mechanisms. Exogenously administered A1M has been shown to alleviate the effects of Hb-induced oxidative stress in rat kidneys. Here we attempted to establish an animal model mimicking the human symptoms at stage two of preeclampsia by administering species-specific cell-free HbF starting mid-gestation until term, and evaluated the therapeutic effect of A1M on the induced symptoms. Female pregnant rabbits received HbF infusions i.v. with or without A1M every second day from gestational day 20. The HbF-infused animals developed proteinuria and a significantly increased glomerular sieving coefficient in kidney that was ameliorated by co-administration of A1M. Transmission electron microscopy analysis of kidney and placenta showed both intracellular and extracellular tissue damages after HbF-treatment, while A1M co-administration resulted in a significant reduction of the structural and cellular changes. Neither of the HbF-treated animals displayed any changes in blood pressure during pregnancy. In conclusion, infusion of cell-free HbF in the pregnant rabbits induced tissue damage and organ failure similar to those seen in preeclampsia, and was restored by co-administration of A1M. This study provides preclinical evidence supporting further examination of A1M as a potential new therapy for preeclampsia. 相似文献
70.
The validity of the Michaelis–Menten–Briggs–Haldane approximation for single enzyme reactions has recently been improved by
the formalism of the total quasi-steady-state approximation. This approach is here extended to fully competitive systems,
and a criterion for its validity is provided. We show that it extends the Michaelis–Menten–Briggs–Haldane approximation for
such systems for a wide range of parameters very convincingly, and investigate special cases. It is demonstrated that our
method is at least roughly valid in the case of identical affinities. The results presented should be useful for numerical
simulations of many in vivo reactions. 相似文献