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1.
《Journal of molecular biology》2021,433(22):167254
Tau is an intrinsically disordered protein implicated in many neurodegenerative diseases. The repeat domain fragment of tau, tau-K18, is known to undergo a disorder to order transition in the presence of lipid micelles and vesicles, in which helices form in each of the repeat domains. Here, the mechanism of helical structure formation, induced by a phospholipid mimetic, sodium dodecyl sulfate (SDS) at sub-micellar concentrations, has been studied using multiple biophysical probes. A study of the conformational dynamics of the disordered state, using photoinduced electron transfer coupled to fluorescence correlation spectroscopy (PET-FCS) has indicated the presence of an intermediate state, I, in equilibrium with the unfolded state, U. The cooperative binding of the ligand (L), SDS, to I has been shown to induce the formation of a compact, helical intermediate (IL5) within the dead time (∼37 µs) of a continuous flow mixer. Quantitative analysis of the PET-FCS data and the ensemble microsecond kinetic data, suggests that the mechanism of induction of helical structure can be described by a U ↔ I ↔ IL5 ↔ FL5 mechanism, in which the final helical state, FL5, forms from IL5 with a time constant of 50–200 µs. Finally, it has been shown that the helical conformation is an aggregation-competent state that can directly form amyloid fibrils. 相似文献
2.
3.
The phytoplankton of some gravel-pit lakes in Spain 总被引:1,自引:1,他引:0
The phytoplankton communities of thirteen adjacent gravel-pit lakes in the lower Jarama river watershed (Madrid, Spain), were studied during spring mixing and summer stratification.If different seasons, the phytoplankton responded to different environmental factors. During spring, the abundance of SRP (soluble reactive phosphorus) and existence of a certain thermal stability resulted in the development of a greater biomass in some lakes. During summer, however, excessively high temperatures adversely affected the communities of the warmer lakes. At the species level, the responses were diverse; ordination techniques enabled us to group them.Some similarities were observed in phytoplankton composition between lakes, possibly due to local dispersion between adjacent lakes (frequented by abundant waterfowl). 相似文献
4.
We designed an Integrated Media Preparation System (IMPS) for continuous, on-line preparation of cell culture media and delivery to intermediate storage vessels or directly to a bioreactor. Key components of the IMPS include: a high precision, continuous fluid mixing device; formulation-specific liquid medium concentrates; validated process controls and membrane filtration; and automated dispensing into large volume flexible plastic containers. The IMPS system is designed to produce sterile, single-strength liquid medium from common raw materials at a delivery rate of 1000–3000 liters per hour and will manufacture homogenous batches from several thousand liters to over 60,000 liters. Fortified nutrient media prepared from multi-component 50X concentrates have been demonstrated to accelerate bioreactor seed chains, increase product yield, and reduce the overall manufacturing cost of nutrient medium. A productivity matrix will analyze the fully-loaded costs and contrast alternative methods for media preparation against projected biological yield.Abbreviations IMPS
Integrated Media Preparation System
- 50X
Nutrient fluid components formulated at fifty-fold final use concentration
- 1X
Nutrient fluid formulated at final, single-strength use concentration
- cGMP
Current Good Manufacturing Practices
- SCADA
Supervisory Control and Data Acquisition
- PLC
Process Logic Controller
- LTI
Life Technologies, Inc.
- WFI
Water for Injection
- CIP
Clean in place
- SIP
Sterilize in place
- HPLC
High performance liquid chromatography
- DMEM
Dulbecco's Modified Eagle's Medium 相似文献
5.
Nienow AW Langheinrich C Stevenson NC Emery AN Clayton TM Slater NK 《Cytotechnology》1996,22(1-3):87-94
Because of concern for cell damage, very low agitation energy inputs have been used in industrial animal cell bioreactors, typical values being two orders of magnitude less than those found in bacterial fermentations. Aeration rates are also very small. As a result, such bioreactors might be both poorly mixed and also unable to provide the higher oxygen up-take rates demanded by more intensive operation. This paper reports experimental studies both of K
L
a and of mixing (via pH measurements) in bioreactors up to 8 m3 at Wellcome and of scaled down models of such reactors at Birmingham. Alongside these physical measurements, sensitivity of certain cell lines to continuously controlled dO2 has been studied and the oxygen up-take rates measured in representative growth conditions. An analysis of characteristic times and mixing theory, together with other recent work showing that more vigorous agitation and aeration can be used especially in the presence of Pluronic F-68, indicates ways of improving their performance. pH gradients offer a special challenge. 相似文献
6.
Recent palaeolimnology of three shallow Danish lakes 总被引:4,自引:0,他引:4
Recent eutrophication histories of three shallow lakes (mean depths <3 m) were studied using palaeolimnological methods. Freeze-cores were dated using 210Pb, 137Cs, 134Cs and 241Am. Resultant chronologies were problematical at two sites (Væng Sø and Vesterborg Sø) due to sediment-water interface mixing, indicated by uniform 137Cs profiles over the surface 20–30 cm. Sediments at Langesø and Vesterborg Sø have a high carbonate content, which together with the high mixing rate have resulted in diatom dissolution below 30 cm at Vesterborg Sø. Diatom stratigraphy indicates relatively small biological changes at both Væng Sø and Langesø both lakes have been eutrophic for at least the last 150–200 years. Væng Sø is dominated by planktonic diatoms together with high percentages of benthic Fragilaria spp., and Langesø by planktonic diatoms, especially Cyclostephanos dubius and Stephanodiscus parvus.Epilimnetic phosphorus concentrations were inferred using weighted averaging; at Væng Sø the diatom-inferred TP (DI-TP) concentrations were close to observed values in the early 1980s, but failed to record the post-biomanipulation decrease to 55 g l-1 after 1988, presumably due to the smoothing of the sediment record by resuspension and mixing. At Langesø two increases in DI-TP suggest a two-phase enrichment of the lake, initial eutrophication 1880, and hypertrophy after 1950. The recent DI-TP values are lower than the annual observed values, and reasons for this are discussed. Despite problems associated with sediment mixing, the sediment records of these shallow systems can still be used for monitoring and for environmental reconstructions. 相似文献
7.
Plant secondary compounds can prevent feeding by phytophagous insects or, if ingested, can be poisonous to them. Less attention has been paid to the additional effects they have on feeding behavior when they are only weakly deterrent or not deterrent at all. Experiments were carried out on the generalist grass-hopperSchistocerca americana. Individuals were presented either with two cakes of high-quality artificial food with a single deterrent compound added or with two cakes, each with a different added deterrent compound. The deterrents consisted of single plant secondary compounds that were either marginally or strongly deterrent. There were profound differences in feeding behavior between those individuals given identical and those given different cake types, including longer feeding bouts on single cakes when a choice of different cake types was available. The behavioral effects demonstrate that the presence of secondary compounds in one food can influence the patterns of feeding on other available foods and suggest that such chemicals could impact foraging activities in a complex manner. 相似文献
8.
R N Cox R R Kaldany P W Brandt B Ferren R A Hudson A Karlin 《Analytical biochemistry》1984,136(2):476-486
A continuous-flow technique is described in which a photoaffinity label, membrane rich in acetylcholine receptor, and various effectors are rapidly mixed, passed through a delay tube, through a tube in which they are irradiated, and are collected in a tube containing quencher. Delay times as short as 20 ms between mixing and photolysis are achievable. Because the flow is continuous, milliliter volumes of membrane can be labeled in a single run, which is convenient for the analysis of both the functional effects and sites of photolabeling. Using this technique, we have found that receptor in its transitory, active state, in which the channel is open, is more susceptible to photolabeling by the noncompetitive inhibitor analog [3H] quinacrine azide than is receptor in either its resting or desensitized states, in which the channel is closed. This technique should prove generally useful for the photolabeling of transient conformational states of macromolecules. 相似文献
9.
G. D. Haffner 《Hydrobiologia》1977,54(3):225-231
Seston size distribution was determined with a Coulter Counter to investigate seston interaction and its effects on phytoplankton
production. Spatial distribution patterns of the non-productive component of the seston were as complex as those commonly
associated with algae. Seston might interact influencing production by changes in optical depth, abrasive actions and nutrient
adsorption. 相似文献
10.
The mixing of various molecular species of phosphatidylglycerol and phosphatidylcholine differing in their acyl chain lengths has been studied both in monolayers (π, Δ), and in water dispersions (fluorescence polarization) with varying pH and ionic strength of the aqueous phase and in the presence of the divalent cations Mg2+ and Ca2+. In dilauroylphosphatidylglycerol/dipalmitoylphosphatidylcholine mixtures, both in monolayers and in water dispersions, no phase separation was detected at pH 2.9 where phosphatidylglycerol was protonated. With dipalmitoylphosphatidylglycerol/dipalmitoylphosphatidylcholine mixtures, in monolayers and at the same pH, no phase separation was detected for surface pressures below . In monolayers, and under ionic conditions such that phosphatidylglycerol was ionized (pH 5.6, 10 mM NaCl) miscibility was observed with dilauroylphosphatidylglycerol and dipalmitoylphosphatidylcholine and also with dipalmitoylphosphatidylglycerol and dilauroylphosphatidylcholine. Varying the ionic strength did not alter the miscibility of these lipids. The divalent cations Mg2+ and Ca2+ did not modify that of dilauroylphosphatidylglycerol with dilauroylphosphatidylcholine or with dipalmitoylphosphatidylcholine. Both in monolayers and in water dispersions, dipalmitoylphosphatidylglycerol and dilauroylphosphatidylcholine appeared to be at least partly miscible, in the presence of magnesium. Only in the presence of calcium and at high surface pressure might the monolayer data account for phase separation between these two lipids. The data presented demonstrate the existence of strong cohesive forces between phosphatidylcholine and phosphatidylglycerol with a marked influence of the former on the physical state of the latter. From an analysis of the Δ data, it is suggested that intrafacial hydrogen bonds may play a significant role in stabilizing phosphatidylcholine/phosphatidylglycerol mixtures. 相似文献