An innovative and automated method for the at-line monitoring of secreted protein was developed by harnessing a Surface Plasmon Resonance-based biosensor to a bioreactor. The proof of concept was ...performed by following at-line the relative concentration of a secreted protein produced by transient transfection of mammalian cells in a bioreactor. Our results suggest that our approach can be readily applied to the at-line determination of both protein concentration and bioactivity. Our experimental setup and strategy can thus satisfy the needs related to the development of novel bioprocess control protocols in the context of the new process analytical technology that arises in the biopharmaceutical industry. Biotechnol. Bioeng. 2008;100: 184-188.
We here describe a scheme of spatially modulated surface plasmon resonance (SPR) polarimetry that enables to combine ultra-high phase sensitivity with good signal-to-noise background. The proposed ...approach uses spatial modulation of s-polarized component by birefringent elements and the extraction of phase-polarization information by Fourier-transform methods. This scheme was tested for monitoring the interactions between an antibody and its biological partner. Our experimental data, collected by amplitude-sensitive and phase-sensitive polarimetry demonstrate that the latter scheme provides at least one order of magnitude improvement in terms of detection limit.
Expanded bed adsorption (EBA) is an efficient protein purification process reducing time and steps of downstream processing (DSP) since nonclarified culture media can be processed directly without ...prior treatments such as filtration or centrifugation. However, cells and debris can interact with the adsorbent and affect bed stability as well as purification performance. To optimize EBA operating conditions these biomass/adsorbent interactions have to be understood and characterized. The adsorption of Human Embryonic Kidney cells (HEK 293) on unprimed and nickel‐primed metal affinity adsorbent was studied in a closed loop EBA setup. With the unprimed adsorbent, the overall level of interaction observed was nonsignificant. With the nickel‐primed adsorbent and an initial cell concentration ranging from 0.08 × 106 to 0.2 × 106 cells/mL, biomass/adsorbent interaction was found to be moderate and the adsorption apparent first‐order kinetic rate constant was determined to be k = 0.009 to 0.011 min‐1.
Renewed interest in the measurement of cellular K(+) effluxes has been prompted by the observation that potassium plays an active and important role in numerous key cellular events, in particular ...cell necrosis and apoptosis. Although necrosis and apoptosis follow different pathways, both induce intracellular potassium effluxes. Here, we report the use of potassium-selective microelectrodes located in a microfluidic platform for cell culture to monitor and quantify such effluxes in real time. Using this platform, we observed and measured the early signs of cell lysis induced by a modification of the extracellular osmolarity. Furthermore, we were able to quantify the number of dying cells by evaluating the extracellular potassium concentration. A comparison between the potentiometric measurement with a fluorescent live-dead assay performed under similar conditions revealed the delay between potassium effluxes and cell necrosis. These results suggest that such platforms may be exploited for applications, such as cytotoxicological screening assays or tumor cell proliferation assays, by using extracellular K(+) as cell death marker.
To investigate whether floral resource quality impacts on bumble bee floral choices, we determined the pollen foraging constancy and floral choices of four bumble bee species commonly occurring in ...peaty, wet meadows in South Belgium. We subsequently analyzed the chemical contents of pollen and nectar, as well as the nectar production of the major host plant species. Individuals of B. lapidarius and B. pascuorum collected high-quality pollen (i.e., having high essential amino acid and phytosterol content) on Comarum palustre and Trifolium pratense, whereas individuals of B. terrestris s.l. and B. hypnorum enlarged their diet breadth to less valuable pollen resources (Cirsium palustre and Valeriana repens). Since Persicaria bistorta and Comarum palustre offer abundant and concentrated nectar, these plant species might represent major nectar sources for bumble bee species in peaty, wet meadows. The present study demonstrated the role of pollen composition on differences in foraging strategies among bumble bee species.
Several bee species are experiencing significant population declines. As bees exclusively rely on pollen for development and survival, such declines could be partly related to changes in their host ...plant abundance and quality. Here, we investigate whether generalist bumblebee species, with stable population trends over the past years, adapted their diets in response to changes in the distribution and chemical quality of their pollen resources. We selected five common species of bumblebee in NW Europe for which we had a precise description of their pollen diet through two time periods (‘prior to 1950’ and ‘2004–2005’). For each species, we assessed whether the shift in their pollen diet was related with the changes in the suitable area of their pollen resources. Concurrently, we evaluated whether the chemical composition of pollen resources changed over time and experimentally tested the impact of new major pollen species on the development of B. terrestris microcolonies. Only one species (i.e. B. lapidarius) significantly included more pollen from resources whose suitable area expanded. This opportunist pattern could partly explain the expansion of B. lapidarius in Europe. Regarding the temporal variation in the chemical composition of the pollen diet, total and essential amino acid contents did not differ significantly between the two time periods while we found significant differences among plant species. This result is driven by the great diversity of resources used by bumblebee species in both periods. Our bioassay revealed that the shift to new major pollen resources allowed microcolonies to develop, bringing new evidence on the opportunist feature of bumblebee in their diets. Overall, this study shows that the response to pollen resource drift varies among closely related pollinators, and a species‐rich plant community ensures generalist species to select a nutrient‐rich pollen diet.
Debris flows affect people and infrastructure around the world, and as a result, many numerical models and modelling approaches have been developed to simulate their impacts. Observations from ...instrumented debris-flow channels show that variability in inflow depth, velocity, and discharge in real debris flows is much higher than what is typically used in numerical simulations. However, the effect of this natural variability on numerical model outputs is not well known. In this study, we examine the effects of using complex inflow time series within a single-phase runout model utilizing a Voellmy flow-resistance model. The interactions between model topography and flow resistance were studied first using a simple triangular hydrograph, which showed that simulated discharges change because of local slopes and Voellmy parameters. Next, more complex inflows were tested using time series based on 24 real debris-flow hydrographs initiated from three locations. We described a simple method to scale inflow hydrographs by defining a target event volume and maximum allowable peak discharge. The results showed a large variation in simulated flow depths and velocities arising from the variable inflow. The effects of variable-inflow conditions were demonstrated in simulations of two case histories of real debris flows, where the variation in inflow leads to significant variations in the simulation outputs. The real debris-flow hydrographs were used to provide an indication of the range of impacts that may result from the natural variability in inflow conditions. These results demonstrate that variation in inflow conditions can lead to reasonable estimates of the potential variation in impacts.
Bumblebees are the predominant wild pollinators for many plant species in temperate regions. A bumblebee colony requires pollen and nectar throughout its lifetime, but degraded and fragmented ...habitats may have gaps in the temporal and spatial continuity of floral resources.
Heathlands are open biotopes that provide favourable habitat for bumblebees like Bombus jonellus, a declining species in Belgium. In heathlands, ericaceous species are the main plants that provide pollen and nectar for bumblebees. Although the nectar composition of ericaceous species has been previously studied, data on pollen composition remain scarce.
We examined bumblebee diets (composition of their pollen loads) in Belgian heathlands over the course of a colony lifetime to assess the fidelity of bumblebees for ericaceous species. We compared nutritional values by investigating the chemical composition (amino acids, polypeptides and sterols) of the pollen of the ericaceous and dominant non‐ericaceous species present in pollen loads. No relationship was detected between the abundance of a particular plant species in bumblebee loads and its pollen composition.
The successive flowering periods and the nutritional quality of pollen of ericaceous species offer valuable resources for bumblebees. Ericaceous species represent a large part of bumblebee diets in heathlands, especially in early spring and late summer when the diversity of other flowering species was low.
Bumblebee pollen loads also contained non‐ericaceous flowering species that grow outside heathlands. Thus, land planning must incorporate conservation strategies for the different elements of the landscape matrix, including heathlands, peatlands, meadows and margins.
This paper describes the calculation of the statistical weights and the Einstein
A-coefficients for the 39 molecules and their associated isotopologues/isotopomers currently present in the ...line-by-line portion of the
HITRAN database. Calculation of the Einstein
A-coefficients was carried out using the
HITRAN line intensities and the necessary statistical weights. The Einstein
A-coefficient and the statistical weights of the upper and lower levels of the transition were added in the new format of the line parameters for the most recent edition of the
HITRAN database.
•The HITRAN2020 molecular spectroscopic database and its validations are presented.•Extended line-by-line coverage to 55 molecules, with new isotopologues included.•Increased spectral and dynamic ...ranges for multiple molecules.•Quality and amount of spectral parameters (including sophisticated line shapes) is increased.•Updates to cross sections, CIA, software tools & auxiliary data also described.
The HITRAN database is a compilation of molecular spectroscopic parameters. It was established in the early 1970s and is used by various computer codes to predict and simulate the transmission and emission of light in gaseous media (with an emphasis on terrestrial and planetary atmospheres). The HITRAN compilation is composed of five major components: the line-by-line spectroscopic parameters required for high-resolution radiative-transfer codes, experimental infrared absorption cross-sections (for molecules where it is not yet feasible for representation in a line-by-line form), collision-induced absorption data, aerosol indices of refraction, and general tables (including partition sums) that apply globally to the data. This paper describes the contents of the 2020 quadrennial edition of HITRAN. The HITRAN2020 edition takes advantage of recent experimental and theoretical data that were meticulously validated, in particular, against laboratory and atmospheric spectra. The new edition replaces the previous HITRAN edition of 2016 (including its updates during the intervening years).
All five components of HITRAN have undergone major updates. In particular, the extent of the updates in the HITRAN2020 edition range from updating a few lines of specific molecules to complete replacements of the lists, and also the introduction of additional isotopologues and new (to HITRAN) molecules: SO, CH3F, GeH4, CS2, CH3I and NF3. Many new vibrational bands were added, extending the spectral coverage and completeness of the line lists. Also, the accuracy of the parameters for major atmospheric absorbers has been increased substantially, often featuring sub-percent uncertainties. Broadening parameters associated with the ambient pressure of water vapor were introduced to HITRAN for the first time and are now available for several molecules.
The HITRAN2020 edition continues to take advantage of the relational structure and efficient interface available at www.hitran.org and the HITRAN Application Programming Interface (HAPI). The functionality of both tools has been extended for the new edition.
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