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Proceedings of the National Academy of... Jul 2023The serotonin transporter (SERT) is a member of the SLC6 neurotransmitter transporter family that mediates serotonin reuptake at presynaptic nerve terminals. SERT is the...
The serotonin transporter (SERT) is a member of the SLC6 neurotransmitter transporter family that mediates serotonin reuptake at presynaptic nerve terminals. SERT is the target of both therapeutic antidepressant drugs and psychostimulant substances such as cocaine and methamphetamines, which are small molecules that perturb normal serotonergic transmission by interfering with serotonin transport. Despite decades of studies, important functional aspects of SERT such as the oligomerization state of native SERT and its interactions with potential proteins remain unresolved. Here, we develop methods to isolate SERT from porcine brain (pSERT) using a mild, nonionic detergent, utilize fluorescence-detection size-exclusion chromatography to investigate its oligomerization state and interactions with other proteins, and employ single-particle cryo-electron microscopy to elucidate the structures of pSERT in complexes with methamphetamine or cocaine, providing structural insights into psychostimulant recognition and accompanying pSERT conformations. Methamphetamine and cocaine both bind to the central site, stabilizing the transporter in an outward open conformation. We also identify densities attributable to multiple cholesterol or cholesteryl hemisuccinate (CHS) molecules, as well as to a detergent molecule bound to the pSERT allosteric site. Under our conditions of isolation, we find that pSERT is best described as a monomeric entity, isolated without interacting proteins, and is ensconced by multiple cholesterol or CHS molecules.
Topics: Animals; Swine; Serotonin Plasma Membrane Transport Proteins; Cryoelectron Microscopy; Detergents; Serotonin; Central Nervous System Stimulants; Cocaine; Methamphetamine
PubMed: 37436958
DOI: 10.1073/pnas.2304602120 -
MethodsX Jun 2024Surfactants are indispensable in industrial applications today due to their wetting, emulsifying, dispersing, cleansing, and detergent properties. The use of surfactants...
Surfactants are indispensable in industrial applications today due to their wetting, emulsifying, dispersing, cleansing, and detergent properties. The use of surfactants extends from the cosmetic industry to the petroleum industry and beyond. Their characteristics and effectiveness can be assessed through various standardized tests, and based on these methods, their applications can be determined. However, there is a lack of a universally applicable testing method for one crucial and complex property: the detergent effect. The detergent effect refers to the removal of unwanted contaminants from a solid surface. However, cleaning is not solely attributed to the surfactant but to the appropriate combination of various factors, whose synergistic effect reduces surface contamination. The most significant factors influencing detergent effect include the characteristics and nature of the contaminants, properties of the cleaning solution (surfactant concentration and composition, water hardness, enzymes, etc.), temperature, washing time, and hydrodynamic conditions. Additionally, the presence of electrolytes, pH of the cleaning solution, and detergent foaming properties may also play important roles. Our goal was to develop a detergent effect testing methodology that is not specific to any particular application domain but offers a straightforward and easy-to-implement solution for comparing the detergent effect of various types of surfactants.•The study presents a method for determining detergent effect of surfactants.•The method is universal and suitable for the evaluation of any type of surfactant.•The method is low-cost and easy to perform.
PubMed: 38799038
DOI: 10.1016/j.mex.2024.102755 -
The Journal of Hospital Infection Oct 2023Transmission of infections via contaminated endoscopes is a common problem. Manual cleaning, using at least a detergent, is an important step in endoscope processing and...
BACKGROUND
Transmission of infections via contaminated endoscopes is a common problem. Manual cleaning, using at least a detergent, is an important step in endoscope processing and should be performed as soon as possible to avoid drying of organic residues that might interfere with high-level disinfection and promote biofilm formation.
AIM
To assess the efficacy of two detergent-disinfectants, enzymatic and non-enzymatic, and of an enzymatic detergent used during the manual cleaning against a Klebsiella pneumoniae biofilm.
METHODS
A 24 h biofilm statically formed in a Tygon tube was exposed to detergent-disinfectants at 20 °C and 35 °C for 10 mn, and to enzymatic detergent at 45 °C for 60 mn. The logarithmic reduction in bacteria in the Tygon tube and the number of bacteria in the product supernatant were calculated.
FINDINGS
Biofilm formation was reproducible between assays. After exposure to detergent-disinfectants, the logarithmic reduction was between 6.32 and 6.71 log cfu/cm in the Tygon tubes. No bacteria were found in their supernatants. Results in the detergent-disinfectant group were not affected by the exposure temperature or the addition of enzymes. No decrease in the bacterial load was observed in the Tygon tubes after exposure to the enzymatic detergent. Bacteria were found in its supernatant.
CONCLUSION
These results show the importance of the choice of products used during the manual cleaning phase. They also show the potential benefit of combining detergent and disinfectant activity to decrease the bacterial load during the manual cleaning step of endoscope processing.
Topics: Humans; Disinfectants; Klebsiella pneumoniae; Detergents; Disinfection; Endoscopes; Biofilms
PubMed: 37487794
DOI: 10.1016/j.jhin.2023.04.019 -
Clinical Drug Investigation Dec 2023Phase II studies on tauro-urso-deoxycholic acid (TUDCA) raised the promise of safety and efficacy in patients with amyotrophic lateral sclerosis, a currently incurable... (Review)
Review
Phase II studies on tauro-urso-deoxycholic acid (TUDCA) raised the promise of safety and efficacy in patients with amyotrophic lateral sclerosis, a currently incurable and devastating disease. We review the available evidence on the efficacy and safety of TUDCA, administered alone or in combination, by analyzing and comparing published and ongoing studies on amyotrophic lateral sclerosis. Two independent phase II studies (using TUDCA solo or combined with sodium phenylbutyrate) showed similar efficacy in slowing disease progression measured by functional scales. One open-label follow-up TUDCA+sodium phenylbutyrate study suggested a benefit on survival. Two subsequent phase III studies with TUDCA (solo or combined with sodium phenylbutyrate) have been initiated and are currently ongoing. Their completion is expected by the end of 2023 and beginning of 2024. Evidence collected by phase II studies indicates that there are no safety concerns in patients with amyotrophic lateral sclerosis. The efficacy shown in phase II studies was considered sufficient to grant approval in some countries but not in others, owing to discrepant views on the strength of evidence. It will be necessary to wait for the results of ongoing phase III studies to attain a full appreciation of these data.
Topics: Humans; Amyotrophic Lateral Sclerosis; Phenylbutyrates; Taurochenodeoxycholic Acid
PubMed: 37973672
DOI: 10.1007/s40261-023-01324-0 -
Environmental Toxicology and Chemistry Jul 2023Certain emerging pollutants are among the most widely used chemicals globally, causing widespread concern in relation to their use in products devoted to cleaniness and... (Review)
Review
Certain emerging pollutants are among the most widely used chemicals globally, causing widespread concern in relation to their use in products devoted to cleaniness and asepsis. Nonylphenol ethoxylate (NPEOn) is one such contaminant, along with its degradation product, nonylphenol, an active ingredient presents in nonionic surfactants used as herbicides, cosmetics, paints, plastics, disinfectants, and detergents. These chemicals and their metabolites are commonly found in environmental matrices. Nonylphenol and NPEOn, used, are particularly concerning, given their role as endocrine disruptors chemical and possible neurotoxic effects recorded in several biological models, primarily aquatic organisms. Limiting and detecting these compounds remain of paramount importance. The objective of the present review was to evaluate the toxic effects of nonylphenol and NPEOn in different biological models. Environ Toxicol Chem 2023;42:1439-1450. © 2023 SETAC.
Topics: Endocrine Disruptors; Phenols; Models, Biological; Water Pollutants, Chemical
PubMed: 37057841
DOI: 10.1002/etc.5633 -
Allergy Jan 2024Epithelial barrier impairment is associated with many skin and mucosal inflammatory disorders. Laundry detergents have been demonstrated to affect epithelial barrier...
BACKGROUND
Epithelial barrier impairment is associated with many skin and mucosal inflammatory disorders. Laundry detergents have been demonstrated to affect epithelial barrier function in vitro using air-liquid interface cultures of human epithelial cells.
METHODS
Back skin of C57BL/6 mice was treated with two household laundry detergents at several dilutions. Barrier function was assessed by electric impedance spectroscopy (EIS) and transepidermal water loss (TEWL) measurements after the 4 h of treatments with detergents. RNA sequencing (RNA-seq) and targeted multiplex proteomics analyses in skin biopsy samples were performed. The 6-h treatment effect of laundry detergent and sodium dodecyl sulfate (SDS) was investigated on ex vivo human skin.
RESULTS
Detergent-treated skin showed a significant EIS reduction and TEWL increase compared to untreated skin, with a relatively higher sensitivity and dose-response in EIS. The RNA-seq showed the reduction of the expression of several genes essential for skin barrier integrity, such as tight junctions and adherens junction proteins. In contrast, keratinization, lipid metabolic processes, and epidermal cell differentiation were upregulated. Proteomics analysis showed that the detergents treatment generally downregulated cell adhesion-related proteins, such as epithelial cell adhesion molecule and contactin-1, and upregulated proinflammatory proteins, such as interleukin 6 and interleukin 1 beta. Both detergent and SDS led to a significant decrease in EIS values in the ex vivo human skin model.
CONCLUSION
The present study demonstrated that laundry detergents and its main component, SDS impaired the epidermal barrier in vivo and ex vivo human skin. Daily detergent exposure may cause skin barrier disruption and may contribute to the development of atopic diseases.
Topics: Humans; Mice; Animals; Detergents; Mice, Inbred C57BL; Skin; Epidermis; Inflammation
PubMed: 37766519
DOI: 10.1111/all.15891 -
Journal of Chromatography. A Jul 2023Cloud-point extraction (CPE) is a pre-treatment technique for the extraction and preconcentration of different chemical compounds, such as metal ions, pesticides, drugs,...
Cloud-point extraction (CPE) is a pre-treatment technique for the extraction and preconcentration of different chemical compounds, such as metal ions, pesticides, drugs, phenols, vitamins etc., from various samples. CPE is based on the phenomenon of two phases (micellar and aqueous) forming after the heating of an aqueous isotropic solution of a non-ionic or zwitterionic surfactant above the cloud-point temperature. If analytes are added to the surfactant solution under suitable conditions, they should be extracted into the micellar phase, also called the surfactant-rich phase. Recently, the traditional CPE procedure is being increasingly replaced by improved CPE procedures. In this study, recent advances in CPE over the last three years (2020 - 2022), including the application of various innovative approaches, are reviewed. In addition to the basic principle of CPE, alternative extraction media in CPE, CPE supported by various auxiliary energies, a different modified CPE procedure and the use nanomaterials and solid-phase extraction in combination with CPE are presented and discussed. Finally, some future trends for improved CPE are presented.
Topics: Octoxynol; Surface-Active Agents; Water; Solid Phase Extraction; Phenols
PubMed: 37207414
DOI: 10.1016/j.chroma.2023.464053 -
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 -
Tau-RNA complexes inhibit microtubule polymerization and drive disease-relevant conformation change.Brain : a Journal of Neurology Aug 2023Alzheimer's disease and related disorders feature neurofibrillary tangles and other neuropathological lesions composed of detergent-insoluble tau protein. In recent...
Alzheimer's disease and related disorders feature neurofibrillary tangles and other neuropathological lesions composed of detergent-insoluble tau protein. In recent structural biology studies of tau proteinopathy, aggregated tau forms a distinct set of conformational variants specific to the different types of tauopathy disorders. However, the constituents driving the formation of distinct pathological tau conformations on pathway to tau-mediated neurodegeneration remain unknown. Previous work demonstrated RNA can serve as a driver of tau aggregation, and RNA associates with tau containing lesions, but tools for evaluating tau/RNA interactions remain limited. Here, we employed molecular interaction studies to measure the impact of tau/RNA binding on tau microtubule binding and aggregation. To investigate the importance of tau/RNA complexes (TRCs) in neurodegenerative disease, we raised a monoclonal antibody (TRC35) against aggregated tau/RNA complexes. We showed that native tau binds RNA with high affinity but low specificity, and tau binding to RNA competes with tau-mediated microtubule assembly functions. Tau/RNA interaction in vitro promotes the formation of higher molecular weight tau/RNA complexes, which represent an oligomeric tau species. Coexpression of tau and poly(A)45 RNA transgenes in Caenorhabditis elegans exacerbates tau-related phenotypes including neuronal dysfunction and pathological tau accumulation. TRC35 exhibits specificity for Alzheimer's disease-derived detergent-insoluble tau relative to soluble recombinant tau. Immunostaining with TRC35 labels a wide variety of pathological tau lesions in animal models of tauopathy, which are reduced in mice lacking the RNA binding protein MSUT2. TRC-positive lesions are evident in many human tauopathies including Alzheimer's disease, progressive supranuclear palsy, corticobasal degeneration and Pick's disease. We also identified ocular pharyngeal muscular dystrophy as a novel tauopathy disorder, where loss of function in the poly(A) RNA binding protein (PABPN1) causes accumulation of pathological tau in tissue from post-mortem human brain. Tau/RNA binding drives tau conformational change and aggregation inhibiting tau-mediated microtubule assembly. Our findings implicate cellular tau/RNA interactions as modulators of both normal tau function and pathological tau toxicity in tauopathy disorders and suggest feasibility for novel therapeutic approaches targeting TRCs.
Topics: Humans; Mice; Animals; tau Proteins; Alzheimer Disease; RNA; Neurodegenerative Diseases; Detergents; Polymerization; Tauopathies; Brain; RNA, Messenger; Caenorhabditis elegans; Microtubules; Poly(A)-Binding Protein I
PubMed: 36732296
DOI: 10.1093/brain/awad032 -
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