ABSTRACT
PURPOSE: Chemical modifications in monoclonal antibodies can change hydrophobicity, charge heterogeneity as well as conformation, which eventually can impact their physical stability. In this study, the effect of the individual charge variants on physical stability and aggregation propensity in two different buffer conditions used during downstream purification was investigated. METHODS: The charge variants were separated using semi-preparative cation exchange chromatography and buffer exchanged in the two buffers with pH 6.0 and 3.8. Subsequently each variant was analysed for size heterogeneity using size exclusion chromatography and dynamic light scattering, conformational stability, colloidal stability, and aggregation behaviour under accelerated stability conditions. RESULTS: Size variants in each charge variant were similar in both pH conditions when analyzed without extended storage. However, conformational stability was lower at pH 3.8 than pH 6.0. All charge variants showed similar apparent melting temperature at pH 6.0. In contrast, at pH 3.8 variants A3, A5, B2, B3 and B4 display lower Tm, suggesting reduced conformational stability. Further, A2, A3 and A5 exhibit reduced colloidal stability at pH 3.8. In general, acidic variants are more prone to aggregation than basic variants. CONCLUSION: Typical industry practice today is to examine in-process intermediate stability with acidic species and basic species taken as a single category each. We suggest that perhaps stability evaluation needs to be performed at specie level as different acidic or basic species have different stability and this knowledge can be used for clever designing of the downstream process to achieve a stable product.
Subject(s)
Antibodies, Monoclonal , Protein Stability , Antibodies, Monoclonal/chemistry , Hydrogen-Ion Concentration , Drug Stability , Protein Conformation , Protein Aggregates , Chromatography, Ion Exchange/methods , Hydrophobic and Hydrophilic Interactions , Chromatography, Gel , Colloids/chemistry , Biological Products/chemistry , Humans , BuffersABSTRACT
Herein, a step-by-step protocol for simultaneous detection of 20 amino acids commonly present in cell culture media is described. The protocol facilitates detection of both primary and secondary amino acids through a two-step precolumn derivatization strategy using ortho-phthalaldehyde and 9-fluorenylmethyl chloroformate as derivatizing agents. The separation of derivatized amino acids with varying hydrophobicity is achieved through reverse-phase chromatography. The amino acids are simultaneously detected in a single workflow through the use of Variable Wavelength Detector at 338 and 262 nm. The protocol is applicable for both mammalian and bacterial cell culture matrices with an option for automation of precolumn derivatization.
Subject(s)
Amino Acids , Biological Products , Animals , Chromatography, High Pressure Liquid/methods , Amino Acids/chemistry , o-Phthalaldehyde/chemistry , Amines , MammalsABSTRACT
Pulmonary vein isolation (PVI) is used for rhythm control in atrial fibrillation (AF). Posterior wall isolation (PWI) is often an adjunct to PVI. Successful PWI is limited by esophageal location, epicardial bridging signals, tissue thickness, and mapping catheter resolution. High-density grid mapping catheters can assist with PWI. Here, we report a case of a 71-year-old woman with persistent AF who underwent PVI and PWI with high-density grid mapping catheters, thus demonstrating the use of omnipolar technology in facilitating successful PWI.
ABSTRACT
Global temperatures are expected to increase due to climate change, and heat stress is one of the major limiting factors affecting future agriculture. To identify plant-associated microorganisms which can promote heat stress tolerance in wheat, we have screened several bacteria isolated from etiolated seedlings of the invasive noxious weed Parthenium hysterophorus. One isolate designated as Ph-04 was found to confer enhanced heat stress tolerance in wheat. The 16S rRNA gene sequence analysis showed that Ph-04 isolate shared highest sequence identity with Bacillus paramycoides species of the Bacillus cereus group. Ph-04 treated wheat seeds exhibited enhanced germination, longer coleoptile, radicle and seminal root length than control seedlings when grown in the dark at optimum and high temperatures. Similarly, under autotrophic conditions, Ph-04 treated plants also exhibited enhanced heat stress tolerance with a significant increase in membrane integrity and significantly reduced levels of H2O2 under heat stress compared to control plants. This observed heat stress tolerance is associated with constitutively higher basal levels of proline, and activity of antioxidant enzymes, catalase (CAT) and ascorbate peroxidase (APX) in Ph-04 treated plants grown under unstressed conditions with further increase under heat stress conditions compared to controls. Plant recovery after heat stress also showed that the Ph-04 treated plants exhibited significantly less damage in terms of survival percentage and exhibited better morphological and physiological characteristics compared to control plants. The study proves that invasive weeds can harbour potentially beneficial microorganisms, which can be transferred to non-native crop (host) plants to improve climate resilience characteristics.
ABSTRACT
"Existing computational fluid dynamics studies of blood flows have demonstrated that the lower wall stress and higher oscillatory shear index might be the cause of acceleration in atherogenesis of vascular walls in hemodynamics. To prevent the chances of aneurysm wall rupture in the saccular aneurysm at distal aortic bifurcation, clinical biomagnetic studies have shown that extra-corporeal magnetic fields can be deployed to regulate the blood flow. Motivated by these developments, in the current study a finite element computational fluid dynamics simulation has been conducted of unsteady two-dimensional non-Newtonian magneto-hemodynamic heat transfer in electrically conducting blood flow in a bifurcated artery featuring a saccular aneurysm. The fluid flow is assumed to be pulsatile, non-Newtonian and incompressible. The Carreau-Yasuda model is adopted for blood to mimic non-Newtonian characteristics. The transformed equations with appropriate boundary conditions are solved numerically by employing the finite element method with the variational approach in the FreeFEM++ code. Hydrodynamic and thermal characteristics are elucidated in detail for the effects of key non-dimensional parameters i.e. Reynolds number (Reâ¯=â¯14, 21, 100, 200), Prandtl number (Prâ¯=â¯14, 21) and magnetic body force parameter (Hartmann number) (Mâ¯=â¯0.6, 1.2, 1.5) at the aneurysm and throughout the arterial domain. The influence of vessel geometry on blood flow characteristics i.e. velocity, pressure and temperature fields are also visualized through instantaneous contour patterns. It is found that an increase in the magnetic parameter reduces the pressure but increases the skin-friction coefficient in the domain. The temperature decreases at the parent artery (inlet) and both the distant and prior artery with the increment in the Prandtl number. A higher Reynolds number also causes a reduction in velocity as well as in pressure. The blood flow shows different characteristic contours with time variation at the aneurysm as well as in the arterial segment. The novelty of the current research is therefore to present a combined approach amalgamating the Carreau-Yasuda model, heat transfer and magnetohydrodynamics with complex geometric features in realistic arterial hemodynamics with extensive visualization and interpretation, in order to generalize and extend previous studies. In previous studies these features have been considered separately and not simultaneously as in the current study. The present simulations reveal some novel features of biomagnetic hemodynamics in bifurcated arterial transport featuring a saccular aneurysm which are envisaged to be of relevance in furnishing improved characterization of the rheological biomagnetic hemodynamics of realistic aneurysmic bifurcations in clinical assessment, diagnosis and magnetic-assisted treatment of cardiovascular disease."
Subject(s)
Aorta/physiopathology , Aortic Aneurysm/physiopathology , Hemorheology , Magnetic Fields , Models, Cardiovascular , Blood Flow Velocity , Computer Simulation , Electric Conductivity , Energy Transfer , Finite Element Analysis , Hot Temperature , Humans , Pulsatile Flow , Regional Blood Flow , Stress, Mechanical , Time FactorsABSTRACT
Two-dimensional rheological laminar hemodynamics through a diseased tapered artery with a mild stenosis present is simulated theoretically and computationally. The effect of different metallic nanoparticles homogeneously suspended in the blood is considered, motivated by drug delivery (pharmacology) applications. The Eringen micropolar model has been discussed for hemorheological characteristics in the whole arterial region. The conservation equations for mass, linear momentum, angular momentum (micro-rotation), and energy and nanoparticle species are normalized by employing suitable non-dimensional variables. The transformed equations are solved numerically subject to physically appropriate boundary conditions using the finite element method with the variational formulation scheme available in the FreeFEM++ code. A good correlation is achieved between the FreeFEM++ computations and existing results. The effect of selected parameters (taper angle, Prandtl number, Womersley parameter, pulsatile constants, and volumetric concentration) on velocity, temperature, and micro-rotational (Eringen angular) velocity has been calculated for a stenosed arterial segment. Wall shear stress, volumetric flow rate, and hemodynamic impedance of blood flow are also computed. Colour contours and graphs are employed to visualize the simulated blood flow characteristics. It is observed that by increasing Prandtl number (Pr), the micro-rotational velocity decreases i.e., microelement (blood cell) spin is suppressed. Wall shear stress decreases with the increment in pulsatile parameters (B and e), whereas linear velocity increases with a decrement in these parameters. Furthermore, the velocity decreases in the tapered region with elevation in the Womersley parameter (α). The simulations are relevant to transport phenomena in pharmacology and nano-drug targeted delivery in hematology.
Subject(s)
Models, Cardiovascular , Nanoparticles , Arteries , Blood Flow Velocity , Computer Simulation , Hemodynamics , Pulsatile Flow , Stress, MechanicalABSTRACT
This article presents a two-dimensional theoretical study of hemodynamics through a diseased permeable artery with a mild stenosis and an aneurysm present. The effect of metallic nanoparticles on the blood flow is considered, motivated by drug delivery (pharmacology) applications. Two different models are adopted to mimic non-Newtonian characteristics of the blood flow; the Casson (viscoplastic) fluid model is deployed in the core region and the Sisko (viscoelastic) fluid model employed in the peripheral (porous) region. The revised Buongiorno two-component nanofluid model is utilized for nanoscale effects. The blood is considered to contain a homogenous suspension of nanoparticles. The governing equations are derived by extending the Navier-Stokes equations with linear Boussinesq approximation (which simulates both heat and mass transfer). Natural (free) double-diffusive convection is considered to simulate the dual influence of thermal and solutal buoyancy forces. The conservation equations are normalised by employing appropriate non-dimensional variables. The transformed equations are solved numerically using the finite element method with the variational formulation scheme available in the FreeFEM++ code. A comprehensive mesh-independence study is included. The effect of selected parameters (thermophoresis, Brownian motion, Grashof number, thermo-solutal buoyancy ratio, Sisko parameter ratio, and permeability parameter) on velocity, temperature, nanoparticle concentration, and hemodynamic pressure have been calculated for two clinically important cases of arteries with stenosis and an aneurysm. Skin-friction coefficient, Nusselt number, volumetric flow rate, and resistance impedance of blood flow are also computed. Colour contours and graphs are employed to visualize the simulated blood flow characteristics. It is observed that by increasing the thermal buoyancy parameter, i.e. Grashof number (Gr), the nanoparticle concentration and temperature decrease, whereas velocity increases with an increment in the Brownian motion parameter (Nb). Furthermore, velocity decreases in the peripheral porous region with elevation in the Sisko material ratio (m) and permeability parameter (k'). The simulations are relevant to transport phenomena in pharmacology and nano-drug targeted delivery in haematology.
Subject(s)
Aneurysm/physiopathology , Arteries/physiopathology , Computer Simulation , Hemodynamics/physiology , Hydrodynamics , Nanoparticles/chemistry , Constriction, Pathologic , Friction , Motion , Numerical Analysis, Computer-Assisted , Porosity , Rheology , TemperatureABSTRACT
PURPOSE: Children with chronic conditions experience medical issues over long-term periods of time which can have lasting emotional and social consequences impacting daily life and functioning. Activities and participation outcomes are needed in order to comprehensively assess child-important health in clinical trials. Our objective was to review the extent to which activity and participation outcomes are included in clinical trials of childhood chronic disease and to determine what trial characteristics are associated with their use. METHODS: A review of a large clinical trial registration database (clinicaltrials.gov) was conducted over the 2010 calendar year. The measures used to assess primary and secondary endpoints were coded according to the ICF classification system. Trial characteristics that might be associated with activity and participation outcome use such as sponsorship type, intervention type, health condition, whether the trial was focused on pediatric patients, phase of trial and sample size were also extracted and explored with univariable and multivariable regressions. RESULTS: Four hundred and ninety-nine trials met inclusion criteria, 495 of which had complete information about hypothesized predictors. Only 36 out of 495 trials included an activity and participation outcome as part of the trial evaluation process. Both univariable and multivariable regression models showed that non-drug trials and late phase of trial (phase IV) showed the strongest likelihood with whether a trial would include an activity and participation outcome. DISCUSSION: Most registered clinical trials for children with chronic or ongoing medical conditions do not include a comprehensive approach to health outcomes assessment, especially drug trials and early phase trials. Outcome measures in pediatric clinical trials are lagging relative to World Health Organization standards for comprehensive health evaluation.
Subject(s)
Chronic Disease/psychology , Clinical Trials as Topic , Patient Outcome Assessment , Social Participation , Activities of Daily Living/classification , Child , Chronic Disease/therapy , Humans , International Classification of Functioning, Disability and Health , Patient Selection , Pediatrics , Quality of Life , Regression Analysis , Research Support as Topic , Sickness Impact ProfileABSTRACT
AIM: Our aims were to (1) describe the conceptual basis of popular generic instruments according to World Health Organization (WHO) definitions of functioning, disability, and health (FDH), and quality of life (QOL) with health-related quality of life (HRQOL) as a subcomponent of QOL; (2) map the instruments to the International Classification of Functioning, Disability and Health (ICF); and (3) provide information on how the analyzed instruments were used in the literature. This should enable users to make valid choices about which instruments have the desired content for a specific context or purpose. METHOD: Child health-based literature over a 5-year period was reviewed to find research employing health status and QOL/HRQOL instruments. WHO definitions of FDH and QOL were applied to each item of the 15 most used instruments to differentiate measures of FDH and QOL/HRQOL. The ICF was used to describe the health and health-related content (if any) in those instruments. Additional aspects of instrument use were extracted from these articles. RESULTS: Many instruments that were used to measure QOL/HRQOL did not reflect WHO definitions of QOL. The ICF domains within instruments were highly variable with respect to whether body functions, activities and participation, or environment were emphasized. INTERPRETATION: There is inconsistency among researchers about how to measure HRQOL and QOL. Moreover, when an ICF content analysis is applied, there is variability among instruments in the health components included and emphasized. Reviewing content is important for matching instruments to their intended purpose.