An extracellular protease inhibitor (BTPI-301) of trypsin was purified and characterized from an isolate of Pseudomonas mendocina.
Results
BTPI-301was purified to homogeneity by (NH4)2SO4, precipitation, DEAE Sepharose and CNBr-activated Sepharose chromatography. Homogeneity was proved by native PAGE and SDS-PAGE. The intact molecular mass was 11567 Da by MALDI-TOF analysis. BTPI-301was a competitive inhibitor with a Ki of 3.5 × 10?10 M. It was stable and active at pH 4–12 and also at 4–90 °C for 1 h. Peptide mass fingerprinting by MALDI revealed that the BTPI-301 is a new inhibitor not reported so far with protease inhibitory activity. The pI of the inhibitor was 3.8. The stoichiometry of trypsin-BTPI-301 interaction is 1:1. The inhibitor was specific towards trypsin.
Conclusion
A pH tolerant and thermostable protease inhibitor BTPI-301 active against trypsin was purified and characterized from P. mendocina that could be developed and used as biopreservative as well as biocontrol agent.
We developed nine microsatellite loci using an enriched library method from the genomic DNA of Pratt’s leaf-nosed bat (Hipposideros pratti). These loci were tested on 96 individuals sampled from Sichuan Province, China. The number of alleles per locus varied from
4 to 14. The expected and observed heterozygosity values ranged from 0.513 to 0.886 and from 0.375 to 0.966, respectively.
Three microsatellite loci departed significantly from Hardy–Weinberg equilibrium (HWE). No linkage disequilibrium was found.
These microsatellite loci will be used in studies of conservation genetics in this species.相似文献
To investigate the effects of Procambarus clarkii on macroinvertebrate diversity, we conducted a mesocosm experiment simulating small pools in rice field pads after the rice
season. We hypothesized that crayfish predation would negatively impact macroinvertebrate diversity, and the magnitude of
this impact should vary with the size of P. clarkii. We conducted a short-term mesocosm experiment to determine macroinvertebrate diversity in the presence of three size classes
and in the absence of crayfish, as well as the diet composition of crayfish from the three size classes. At the end of the
experiments, the diet of crayfish was composed of the most available taxa (Culicidae, Chironomus, Tanytarsini and Orthocladinae). These results also show evidence that, in confined areas, crayfish are important predators
of major rice pests such as rice Chironominae larvae. Macroinvertebrate diversity was negatively affected by crayfish presence,
but the effect was inversely proportional to crayfish size. The highest diversity index was obtained in the absence of P. clarkii, and juvenile crayfish significantly reduced macroinvertebrate diversity. Thus, the impact of P. clarkii on aquatic macroinvertebrates is size dependent and may be relevant in small pools formed in rice field pads from early autumn
to late winter. Overall, our findings suggest that the negative effects of P. clarkii on macroinvertebrate diversity may be particularly strong in local natural assemblages confined to puddles of water or small
ponds in wetland areas. 相似文献
The accuracy by which phenotype can be reproduced by genotype potentially is important in determining the stability, environmental
sensitivity, and evolvability of morphology and other phenotypic traits. Because two sides of an individual represent independent
development of the phenotype under identical genetic and environmental conditions, average body asymmetry (or "fluctuating
asymmetry") can estimate the developmental instability of the population. The component of developmental instability not explained
by intrapopulational differences in gene or environment (or their interaction) can be further defined as internal developmental
noise. Surprisingly, developmental noise remains largely unexplored despite its potential influence on our interpretations
of developmental stability, canalization, and evolvability. Proponents of fluctuating asymmetry as a bioindicator of environmental
or genetic stress, often make the assumption that developmental noise is minimal and, therefore, that phenotype can respond
sensitively to the environment. However, biologists still have not measured whether developmental noise actually comprises
a significant fraction of the overall environmental response of fluctuating asymmetry observed within a population. 相似文献
Many studies of biochemical networks have analyzed network topology. Such work has suggested that specific types of network
wiring may increase network robustness and therefore confer a selective advantage. However, knowledge of network topology
does not allow one to predict network dynamical behavior – for example, whether deleting a protein from a signaling network
would maintain the network's dynamical behavior, or induce oscillations or chaos. 相似文献
Atrial fibrillation (AF) is reported to be a less frequent cause of ischemic stroke in China than in Europe and North America, but it is not clear whether this is due to underestimation. Our aim was to define the true frequency of AF-associated stroke, to determine the yield of 6-day Holter ECG to detect AF in Chinese stroke patients, and to elucidate predictors of newly detected AF.
Methods
Patients with acute ischemic stroke or transient ischemic attack (TIA) were enrolled in a prospective, multicenter cohort study of 6-day Holter monitoring within 7 days after stroke onset at 20 sites in China between 2013 and 2015. Independent predictors of newly-detected AF were determined by multivariate analysis.
Results
Among 1511 patients with ischemic stroke and TIA (mean age 63 years, 33.1% women), 305 (20.2%) had either previously known (196, 13.0%) or AF newly-detected by electrocardiography (53, 3.5%) or by 6-day Holter monitoring (56/1262, 4.4%). A history of heart failure (OR?=?4.70, 95%CI, 1.64–13.5), advanced age (OR?=?1.06, 95%CI, 1.04–1.09), NIHSS at admission (OR?=?1.06, 95%CI, 1.02–1.10), blood high density lipoprotein (HDL) (OR?=?1.52, 95%CI, 1.09–2.13), together with blood triglycerides (OR?=?0.64, 95%CI, 0.45–0.91) were independently associated with newly-detected AF.
Conclusions
Contrary to previous reports, AF-associated stroke is frequent (20%) in China if systemically sought. Prolonged noninvasive cardiac rhythm monitoring importantly increases AF detection in patients with recent ischemic stroke and TIA in China. Advanced age, history of heart failure, and higher admission NIHSS and higher level of HDL were independent indicators of newly-detected AF.
Focal adhesion kinase (FAK) functions as a key enzyme in the integrin-mediated adhesion-signalling pathway. Here, we aimed to investigate the effects of FAK on adhesion of human dental pulp (HDP) cells. We transfected lentiviral vectors to silence or overexpress FAK in HDP cells ex vivo. Early cell adhesion, cell survival and focal contacts (FCs)-related proteins (FAK and paxillin) were examined. By using immunofluorescence, the formation of FCs and cytoskeleton was detected, respectively. We found that both adhesion and survival of HDP cells were suppressed by FAK inhibition. However, FAK overexpression slightly inhibited cell adhesion and exhibited no change in cell survival compared with the control. A thick rim of cytoskeleton accumulated and smaller dot-shaped FCs appeared in FAK knockdown cells. Phosphorylation of paxillin (p-paxillin) was inhibited in FAK knockdown cells, verifying that the adhesion was inhibited. Less cytoskeleton and elongated FCs were observed in FAK-overexpressed cells. However, p-paxillin had no significant difference compared with the control. In conclusion, the data suggest that FAK maintains cell adhesion, survival and cytoskeleton formation, but excessive FAK has no positive effects on these aspects. 相似文献
The hyperthermophilic α-amylase from Thermococcus sp. HJ21 does not require exogenous calcium ions for thermostability, and is a promising alternative to commercially available α-amylases to increase the efficiency of industrial processes like the liquefaction of starch. We analyzed the amino acid sequence of this α-amylase by sequence alignments and structural modeling, and found that this α-amylase closely resembles the α-amylase from Pyrococcus woesei. The gene of this α-amylase was cloned in Escherichia coli and the recombinant α-amylase was overexpressed and purified with a combined renaturation-purification procedure. We confirmed thermostability and exogenous calcium ion independency of the recombinant α-amylase and further investigated the mechanism of the independency using biochemical approaches. The results suggested that the α-amylase has a high calcium ion binding affinity that traps a calcium ion that would not dissociate at high temperatures, providing a direct explanation as to why the addition of calcium ions is not required for thermostability. Understanding of the mechanism offers a strong base on which to further engineer properties of this α-amylase for better potential applications in industrial processes. 相似文献
Multipurpose NTFP species typically experience higher harvest demand because of their multiple uses, which, when combined with unsustainable land use practices, may threaten population viability. We assessed local knowledge on the uses, habitat, and population status of Mimusops andongensis and Mimusops kummel, both multipurpose NTFP species in Benin, to promote their valorization and conservation and thus sustain local knowledge on their uses for domestication issues. One hundred households were randomly selected for structured interviews for M. andongensis and 500 for M. kummel. The relationship between age, sex, and ethnic groups and the species uses was assessed using comparison and correspondence analyses. Nearly all organs of the species were used. Both species were mainly exploited for medicinal purposes but also in construction and as firewood. We found similarities in some uses of the species organs, although the species occur in different ecological zones and are used by different ethnic groups. This result should be considered for the valorization of the species. Most informants reported that populations of M. andongensis were decreasing, although some felt that they were increasing, whereas less than one-third said that M. kummel was decreasing. There were strong relationships between gender, age, and ethnic affiliation of the users and the exploited organs of both species. Potential uses exist based on both the past and current uses of the species and in comparison to other countries where they are exploited. Local ethnoecological knowledge and practices will help to valorize and conserve the species. However, further research on the species’ seed germination and propagation ability are also necessary. 相似文献
Lipids are essential for mammalian cells to maintain many physiological functions. Emerging evidence has shown that cancer cells can develop specific alterations in lipid biosynthesis and metabolism to facilitate their survival and various malignant behaviors. To date, the precise role of cellular lipids and lipid metabolism in viral oncogenesis is still largely unclear with only a handful of literature covering this topic to implicate lipid metabolism in oncogenic virus associated pathogenesis. In this review, we focus on the role of lipid biosynthesis and metabolism in the pathogenesis of the Kaposi’s sarcoma-associated herpesvirus, a common causative factor for cancers arising in the immunocompromised settings.
