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Environmental Science and Pollution... Mar 2023At household level, clothes washing has been recognised as an emitter of microplastics (MPs) into the environment and it is supposed that dishwashing is also a source of...
At household level, clothes washing has been recognised as an emitter of microplastics (MPs) into the environment and it is supposed that dishwashing is also a source of MPs, although little attention has been paid so far. In this work, the emission of MPs released from dishwashing procedures at household level has been studied. The effect of different parameters such as time, temperature and type of detergent has been analysed. In addition, the MP content of tap water has been evaluated in order to determine its contribution to the MPs in dishwasher effluent. Results showed that when the dishwasher was operated empty with a pre-wash programme (15 min and room water temperature), between 207 and 427 MPs were released per load (3 L), whereas this value increased notably with an intensive programme (164 min and water at 70 °C) (1025-1370 MPs per load, 15 L), which highlighted the effect of temperature and time on MP release. Additionally, when a polypropylene lunch box was washed, the number of MPs released increased by 14 ± 3 MPs and 166 ± 12 MPs of total. Finally, the influence of the use of detergent with the dishwasher empty and containing lunch boxes has been studied. With detergent, 35-54% more MPs were released from dishwasher accessories, whereas no additional release took place from lunch boxes. This work shows for the first time the important contribution of domestic dishwashing to MP pollution and the environmental benefits of using more environmentally friendly materials in both dishwashing machine accessories and food utensils.
Topics: Microplastics; Plastics; Detergents; Environmental Monitoring; Water Pollutants, Chemical
PubMed: 36701065
DOI: 10.1007/s11356-023-25433-7 -
Biotechnology and Bioengineering Apr 2022Detergent-mediated virus inactivation (VI) provides a valuable orthogonal strategy for viral clearance in mammalian processes, in particular for next-generation...
Detergent-mediated virus inactivation (VI) provides a valuable orthogonal strategy for viral clearance in mammalian processes, in particular for next-generation continuous manufacturing. Furthermore, there exists an industry-wide need to replace the conventionally employed detergent Triton X-100 with eco-friendly alternatives. However, given Triton X-100 has been the gold standard for VI due its minimal impact on protein stability and high inactivation efficacy, inactivation by other eco-friendly detergents and its impact on protein stability is not well understood. In this study, the sugar-based detergent commonly used in membrane protein purification, n-dodecyl-β- d-maltoside was found to be a promising alternative for VI. We investigated a panel of detergents to compare the relative VI efficacy, impact on therapeutic quality attributes, and clearance of the VI agent and other impurities through subsequent chromatographic steps. Detergent-mediated inactivation and protein stability showed comparable trends to low pH inactivation. Using experimental and modeling data, we found detergent-mediated product aggregation and its kinetics to be driven by extrinsic factors such as detergent and protein concentration. Detergent-mediated aggregation was also impacted by an initial aggregation level as well as intrinsic factors such as the protein sequence and detergent hydrophobicity, and critical micelle concentration. Knowledge gained here on factors driving product stability and VI provides valuable insight to design, standardize, and optimize conditions (concentration and duration of inactivation) for screening of detergent-mediated VI.
Topics: Animals; Biological Products; Detergents; Kinetics; Mammals; Octoxynol; Protein Stability; Virus Inactivation
PubMed: 35023152
DOI: 10.1002/bit.28034 -
Methods in Molecular Biology (Clifton,... 2020Small angle neutron scattering (SANS) is a powerful tool for studying the structure of solubilized membrane proteins. It allows describing the general dimension of the...
Small angle neutron scattering (SANS) is a powerful tool for studying the structure of solubilized membrane proteins. It allows describing the general dimension of the membrane protein , evidence conformational changes, and may provide a low-resolution structure at the nm resolution range. This is because SANS can discriminate between the membrane protein and its amphiphilic partner by specific deuteration of the partners and of the buffer. This chapter was written to offer to a scientist aiming to describe a membrane protein structure the basic tools to consider a SANS experiment. It presents the general principle of contrast variation and a bibliographic survey of experimental strategies used for membrane proteins, some basic theoretical background, and a succinct description of the principles of analysis, of the instrumental and sample requirement, and of the practical steps, prior to the experiments, during the experiments and for data analysis.
Topics: Detergents; Deuterium Exchange Measurement; Humans; Membrane Proteins; Neutron Diffraction; Scattering, Small Angle
PubMed: 33582991
DOI: 10.1007/978-1-0716-0724-4_7 -
Journal of Microbiological Methods Dec 2023Leishmaniasis is an infectious disease caused by protozoan species in the genera Leishmania and Endotrypanum. Current antileishmanial drugs are limited due to adverse...
Leishmaniasis is an infectious disease caused by protozoan species in the genera Leishmania and Endotrypanum. Current antileishmanial drugs are limited due to adverse effects, variable efficacy, the development of resistant parasites, high cost, parenteral administration and lack of availability in endemic areas. Therefore, active searching for new antileishmanial drugs has been done for years, mainly by academia. Drug screening techniques have been a challenge since the intracellular localization of Leishmania amastigotes implies that the host cell may interfere with the quantification of the parasites and the final estimation of the effect. One of the procedures to avoid host cell interference is based on its detergent-mediated lysis and subsequent transformation of viable amastigotes into promastigotes, their proliferation and eventual quantification as an axenic culture of promastigotes. However, the use of detergent involves additional handling of cultures and variability. In the present work, cultures of intracellular amastigotes were incubated for 72 h at 26 °C after exposure to the test compounds and the transformation and proliferation of parasites took place without need of adding any detergent. The assay demonstrated clear differentiation of negative and positive controls (average Z´ = 0.75) and 50% inhibitory concentrations of compounds tested by this method and by the gold standard enumeration of Giemsa-stained cultures were similar (p = 0.5002) and highly correlated (r = 0.9707). This simplified procedure is less labor intensive, the probability of contamination and the experimental error are reduced, and it is appropriate for the automated high throughput screening of compounds.
Topics: Animals; Leishmania; Parasites; Drug Evaluation, Preclinical; Detergents; Leishmaniasis; Antiprotozoal Agents
PubMed: 37871728
DOI: 10.1016/j.mimet.2023.106847 -
Current Protocols in Protein Science Aug 2014Well-characterized membrane protein detergent complexes (PDC) that are pure, homogenous, and stable, with minimized excess detergent micelles, are essential for... (Review)
Review
Well-characterized membrane protein detergent complexes (PDC) that are pure, homogenous, and stable, with minimized excess detergent micelles, are essential for functional assays and crystallization studies. Procedural steps to measure the mass, size, shape, homogeneity, and molecular composition of PDCs and their host detergent micelles using size-exclusion chromatography (SEC) with a Viscotek Tetra Detector Array (TDA; absorbance, refractive index, light scattering, and viscosity detectors) are presented in this unit. The value of starting with a quality PDC sample, the precision and accuracy of the results, and the use of a digital benchtop refractometer are emphasized. An alternate and simplified purification and characterization approach using SEC with dual absorbance and refractive index detectors to optimize detergent and lipid concentration while measuring the PDC homogeneity is also described. Applications relative to purification and characterization goals are illustrated as well.
Topics: Chromatography, Gel; Detergents; Membrane Proteins; Micelles; Refractometry
PubMed: 25081744
DOI: 10.1002/0471140864.ps2910s77 -
Advanced Biology Jan 2024Solubilizing extracellular matrix (ECM) materials and transforming them into hydrogels has expanded their potential applications both in vitro and in vivo. In this...
Solubilizing extracellular matrix (ECM) materials and transforming them into hydrogels has expanded their potential applications both in vitro and in vivo. In this study, hydrogels are prepared by decellularization of human placental tissue using detergent and enzymes and by the subsequent creation of a homogenized acellular placental tissue powder (P-ECM). A perfusion-based decellularization approach is employed using detergent and enzymes. The P-ECM with and without gamma irradiation is then utilized to prepare P-ECM hydrogels. Physical and biological evaluations are conducted to assess the suitability of the P-ECM hydrogels for biocompatibility. The decellularized tissue has significantly reduced cellular content and retains the major ECM proteins. Increasing the concentration of P-ECM leads to improved mechanical properties of the P-ECM hydrogels. The biocompatibility of the P-ECM hydrogel is demonstrated through cell proliferation and viability assays. Notably, gamma-sterilized P-ECM does not support the formation of a stable hydrogel. Nonetheless, the use of HCl during the digestion process effectively decreases spore growth and bacterial bioburden. The study demonstrates that P-ECM hydrogels exhibit physical and biological attributes conducive to soft tissue reconstruction. These hydrogels establish a favorable microenvironment for cell growth and the need for investigating innovative sterilization methods.
Topics: Female; Pregnancy; Humans; Hydrogels; Detergents; Placenta; Extracellular Matrix; Biological Assay
PubMed: 37786307
DOI: 10.1002/adbi.202300349 -
The Journal of Physical Chemistry. B Nov 2017Although fundamentally significant in structural, chemical, and membrane biology, the interfacial protein-detergent complex (PDC) interactions have been modestly...
Although fundamentally significant in structural, chemical, and membrane biology, the interfacial protein-detergent complex (PDC) interactions have been modestly examined because of the complicated behavior of both detergents and membrane proteins in aqueous phase. Membrane proteins are prone to unproductive aggregation resulting from poor detergent solvation, but the participating forces in this phenomenon remain ambiguous. Here, we show that using rational membrane protein design, targeted chemical modification, and steady-state fluorescence polarization spectroscopy, the detergent desolvation of membrane proteins can be quantitatively evaluated. We demonstrate that depleting the detergent in the sample well produced a two-state transition of membrane proteins between a fully detergent-solvated state and a detergent-desolvated state, the nature of which depended on the interfacial PDC interactions. Using a panel of six membrane proteins of varying hydrophobic topography, structural fingerprint, and charge distribution on the solvent-accessible surface, we provide direct experimental evidence for the contributions of the electrostatic and hydrophobic interactions to the protein solvation properties. Moreover, all-atom molecular dynamics simulations report the major contribution of the hydrophobic forces exerted at the PDC interface. This semiquantitative approach might be extended in the future to include studies of the interfacial PDC interactions of other challenging membrane protein systems of unknown structure. This would have practical importance in protein extraction, solubilization, stabilization, and crystallization.
Topics: Detergents; Fluorescence Polarization; Hydrophobic and Hydrophilic Interactions; Membrane Proteins; Molecular Dynamics Simulation
PubMed: 29035562
DOI: 10.1021/acs.jpcb.7b08045 -
Biochemical Society Transactions Jun 2016Membrane proteins account for a third of the eukaryotic proteome, but are greatly under-represented in the Protein Data Bank. Unfortunately, recent technological... (Review)
Review
Membrane proteins account for a third of the eukaryotic proteome, but are greatly under-represented in the Protein Data Bank. Unfortunately, recent technological advances in X-ray crystallography and EM cannot account for the poor solubility and stability of membrane protein samples. A limitation of conventional detergent-based methods is that detergent molecules destabilize membrane proteins, leading to their aggregation. The use of orthologues, mutants and fusion tags has helped improve protein stability, but at the expense of not working with the sequence of interest. Novel detergents such as glucose neopentyl glycol (GNG), maltose neopentyl glycol (MNG) and calixarene-based detergents can improve protein stability without compromising their solubilizing properties. Styrene maleic acid lipid particles (SMALPs) focus on retaining the native lipid bilayer of a membrane protein during purification and biophysical analysis. Overcoming bottlenecks in the membrane protein structural biology pipeline, primarily by maintaining protein stability, will facilitate the elucidation of many more membrane protein structures in the near future.
Topics: Animals; Detergents; Humans; Membrane Proteins; Protein Stability; Proteomics; Solubility
PubMed: 27284049
DOI: 10.1042/BST20160049 -
International Journal of Molecular... Mar 2023Proteases are important enzymes that are engaged in a variety of essential physiological functions and have a significant possible use in industrial applications. In...
Proteases are important enzymes that are engaged in a variety of essential physiological functions and have a significant possible use in industrial applications. In this work, we reported the purification and biochemical characterization of a detergent stable, antimicrobial, and antibiofilm potential protease (SH21) produced by CSB55 isolated from Korean fermented vegetable kimchi. SH21 was purified to obtain homogeneity via ammonium sulfate precipitation (40-80%), Sepharose CL-6B, and Sephadex G-75 column. By analyzing the SDS-PAGE and zymogram, it was determined that the molecular weight was around 25 kDa. The enzyme activity was almost completely inhibited in the presence of PMSF and DFP, which indicated that it was a member of the serine protease family. SH21 showed excellent activity with a broad range of pH and temperature, with its maximum pH of 9.0 and temperature of 55 °C. The enzyme had estimated K and V values of 0.197 mg/mL and 1.22 × 10 U/mg, respectively. In addition, it preserved good activity in the presence of different organic solvents, surfactants, and other reagents. This enzyme showed good antimicrobial activity that was evaluated by MIC against several pathogenic bacteria. Furthermore, it exhibited strong antibiofilm activity as determined by MBIC and MBEC assay and degraded the biofilms, which were analyzed by confocal microscopic study. These properties established that SH21 is a potent alkaline protease that can be used in industrial and therapeutic applications.
Topics: Detergents; Endopeptidases; Bacillus; Serine Proteases; Temperature; Bacterial Proteins; Anti-Infective Agents; Hydrogen-Ion Concentration; Enzyme Stability
PubMed: 36982846
DOI: 10.3390/ijms24065774 -
Journal of Dental Hygiene : JDH Feb 2023Detergent pod-related aerodigestive/ocular chemical burns/ingestion poisoning injuries are a group of injuries of concern to parents/guardians of young children as well...
Detergent pod-related aerodigestive/ocular chemical burns/ingestion poisoning injuries are a group of injuries of concern to parents/guardians of young children as well as oral healthcare professionals. Each year detergent pod injuries result in thousands of emergency department (ED) visits and significant oral, eye, airway, and digestive trauma. The purpose of this study was to investigate trends in detergent pod injuries resulting in ED visits in children ages >0 to <18 years from 2016-2020. This cross-sectional study used data collected from the National Electronic Injury Surveillance System (NEISS). A secondary data analysis was performed on the NEISS data from 2016-2020 for children ages >0 to <18 years. Injuries were identified as detergent pod-related oral-aerodigestive/ingestion poisonings, detergent pod-related ocular injuries, or other product-related injuries among children. Time trends and demographic characteristics were analysed by age, sex, and race. There were an estimated 13,176 detergent pod-related oral-aerodigestive injuries/ingestion poisonings and 8,654 detergent pod-related ocular injuries with ED visits during 2016 - 2020. In adjusted logistic regression, white children were more likely to have oral-aerodigestive injuries/ingestion poisonings than black children (=0.0006). Time trend (=0.4358), and sex (=0.3905) failed to reach significance. Children, ages 5 to<18 years, were less likely to have ED visits for oral-aerodigestive injuries/ingestion poisonings, or any detergent pod-related injury than younger children. Children, ages 3 to ≤5 years, were more likely to have detergent pod-related ocular injuries than younger or older children. Time trends for detergent-pod related injuries requiring ED visits remained unchanged from 2016-2020 for oral-aerodigestive injuries/ingestion poisonings. Given the significant health risks associated with detergent pods and the fact that the injuries have not declined, there is a need for improved parent/guardian education and practices in safeguarding detergent pods from children. Dental hygienists are well positioned to provide anticipatory guidance on this safety issue.
Topics: Child; Humans; Adolescent; Child, Preschool; Detergents; Cross-Sectional Studies; Educational Status; Health Personnel
PubMed: 36854577
DOI: No ID Found