The presence of surfactants in natural waters has become a growing environmental concern, as these substances, widely used in detergents, cleaning products, cosmetics, and industrial processes, can cause serious impacts on aquatic ecosystems, including deleterious effects on biota, depletion of dissolved oxygen, and eutrophication. This study presents a battery of ecotoxicological assays with different aquatic organisms to evaluate the acute, chronic, and sublethal toxic effects of anionic surfactants Linear Alkylbenzene Sulfonate (LAS) and Sodium Dodecyl Sulfate (SDS). The assays were performed with Vibrio fischeri bacteria, Daphnia similis crustaceans, Hyalella azteca amphipods, and Biomphalaria glabrata snails. Regarding LAS acute exposure, H. azteca was the most sensitive organism, followed by D. similis, B. glabrata, and V. fischeri, with E(L)C50 between 5.04 and 28.47 mg/L. Among the organisms exposed to SDS, the most sensitive were V. fischeri bacteria, followed by D. similis, H. azteca, and B. glabrata, with E(L)C50 values between 0.62 and 36.87 mg/L. Chronic effects on the reproduction of D. similis exposed to both surfactants were also observed, as well as teratogenic and lethal effects on B. glabrata embryos. The data obtained herein demonstrated the importance of evaluating the toxicity of surfactants to aquatic ecosystems, as these stressors caused different adverse effects in the exposed species.

This paper reports on a plausible mechanism of modulation by anionic phospholipids of KcsA, a model prokaryotic potassium channel. A first conclusion is that the association of anionic phospholipids with KcsA occurs at discrete regions located near both the extracellular and intracellular channel gates. This same behaviour is consistently observed in both detergent micelles and lipid bilayer-based experimental systems, indicating that the interaction does not depend on bulk bilayer characteristics. In the outer gate (i.e., the selectivity filter) anionic lipid binding to the so called non-annular lipid binding sites, drives the selectivity filter into a non-inactivating, conductive conformation, with a fairly high affinity to bind K[+]. Likewise, at the inner gate, the anionic lipid binding site seems to include cationic amino acid residues at the so called M0 helix. Here, interaction with anionic lipids facilitates the untangle of the C-terminal α-helical bundle at more neutral pH values, thus, facilitating inner gate opening. Both of these phenomena seem to occur independently and should favour the so called Conductive/Open state in the proposed gating cycle of the channel, which in the absence of anionic lipids is just a short-lived, transient state. Consequently, it is expected that the interactions of anionic lipid with KcsA should result in a great increase in ion channel activity. Indeed, this is the case, as reported in functional studies from different laboratories. Such an ability of specific lipids to regulate channel function points out to an interest in future developments of lipid-like drugs to control channelopathies.

A novel centrifuge-less cloud-point extraction (CL-CPE) method based on pyrogallol (PG) was developed for the spectrophotometric determination of total vanadium. The method employs a mixed micelle-mediated extraction system comprising the nonionic surfactant Triton X-114 and the ionic liquid Aliquat 336 (A336), which serves as a source of monovalent cations capable of forming an ion pair with the anionic vanadium-PG chelate. The extracted species, (A336+)[VIV(OH)(PG)2], exhibits several absorption maxima (309, 362, and 436 nm), providing enhanced selectivity through appropriate wavelength selection according to the sample matrix. The principal absorption maximum occurs at 309 nm. At a sevenfold preconcentration factor, the method provides high sensitivity at this wavelength, with a molar absorptivity of 3.2 × 107 L mol-1 cm-1, a limit of detection of 0.11 ng mL-1, and a Sandell sensitivity of 1.6 × 10-3 ng cm-2. The applicability of the method was demonstrated through the analysis of drinking water samples, spent vanadium catalyst materials, and vanadium-containing dietary supplements. The method was further evaluated using the RGBfast model, a white analytical chemistry assessment tool that integrates analytical performance, environmental sustainability, and practical and economic efficiency. The evaluation indicated a high overall whiteness score.

Background: Access to safe drinking water is a considerable problem in Malawi. Contaminated water plays a major role in diarrheal disease, particularly for people living with HIV and their families. Water, sanitation, and hygiene (WASH) interventions can reduce the risk of diarrhea among children by up to 50%. However, few published reports describe WASH integration in routine HIV care programming in Africa. Objective: To evaluate a WASH program integrated within the prevention of vertical transmission of HIV (PVTH) care in Lilongwe, Malawi. Methods: The University of North Carolina's Project Malawi, the Proctor & Gamble Company (P&G), and the Malawi Ministry of Health implemented a safe drinking water program from 2008 to 2023, integrated within the national PVTH program in Lilongwe, Malawi. We provided a point-of-use WASH package to pregnant and breastfeeding women (PBFW) with HIV and their families that included soap, sieves, closed-water containers, and P&G water purification packets. We examine program impact by describing trends in childhood diarrhea and retention in care at 33 program facilities and 29 non-program facilities from 2018 to 2022. Findings: Over the program's final 5 years, an estimated 2 million water purification packets were distributed annually. From 2018 to 2022 in program facilities, non-bloody diarrhea rates among children under 5 years remained constant, while bloody-diarrhea rates fell at both program and non-program facilities. A small association was found between facility participation in the program and retention in care among PBFW living with HIV (incidence rate ratio: 1.05, 95% CI: 0.99-1.12), but of borderline statistical significance. Conclusions: In an environment where access to clean water is not assured, we demonstrated the feasibility of delivering a WASH package in HIV care settings over 15 consecutive years. The collaboration between government, academic, and industry partners provides an example for how to integrate WASH within PVTH programming.

This study evaluated the effects of dietary non-fiber carbohydrate to neutral detergent fiber ratio (NFC/NDF) and allicin supplementation on serum electrolytes, urinary biochemical indicators, nitrogen (N) and phosphorus excretion, and fecal microbiota in beef bulls. Forty 12-month-old beef bulls were assigned to a 2 × 2 factorial design based on dietary NFC/NDF (0.75 and 1.02) and allicin supplementation (0 and 0.04% on a DM basis), resulting in four treatment groups with 10 replicates per group. Increasing dietary NFC/NDF decreased serum pH and increased the anion gap (p < 0.05). An interaction between NFC/NDF and allicin supplementation was observed for the serum concentrations of non-ionized calcium and total calcium (p < 0.05), with the highest concentrations detected in cattle fed high-NFC/NDF diets supplemented with allicin. Serum uric acid was elevated by high NFC/NDF (p < 0.05). High dietary NFC/NDF also increased urine pH (p < 0.05). Significant interactions between dietary NFC/NDF and allicin supplementation were observed for allantoin, creatinine, and urease activity (p < 0.05). High NFC/NDF increased N excretion, total N excretion, and N excretion rate (p < 0.01). Additionally, allicin supplementation reduced fecal N excretion and N excretion rate and increased the apparent total-tract digestibility of N (p < 0.05). Fecal microbiota Alpha diversity analysis showed that high NFC/NDF decreased the Chao1, observed features, and Shannon (p < 0.05). At the phylum level, high NFC/NDF increased Pseudomonadota but decreased Cyanobacteriota, and a significant interaction between NFC/NDF and allicin was observed for Spirochaetota (p < 0.05). At the genus level, high NFC/NDF reduced unclassified_UCG-010 but increased Succinivibrio, unclassified_Muribaculaceae, and Methanobrevibacter (p < 0.05). Allicin increased Rikenellaceae_RC9_gut_group abundance (p < 0.05). In conclusion, dietary NFC/NDF and allicin supplementation modulate acid-base balance, N utilization, and fecal microbial composition in beef bulls, providing a scientific basis for improving feed efficiency.

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) is a popular technique for the separation and characterization of proteins. Whereas most proteins are denatured by the anionic detergent SDS, the enzyme polyphenol oxidase (PPO) is activated by low SDS concentrations, provided that the samples are not heated and reduced. Direct activity staining for PPO is therefore possible on SDS-PAGE gels. Color development from ortho-quinone reaction products can be enhanced with coupling reagents such as 4-amino-N,N-diethylaniline (ADA). As demonstrated with Agaricus bisporus (white and brown button mushroom) extracts, multiple PPO forms can be clearly resolved on SDS-PAGE gels. The method described here encompasses sample preparation, electrophoresis, and activity staining. It is a rapid, simple, and high-resolution technique that enables the comparison of PPO forms across several samples without the need for protein purification.

Commonly available alternative methods for the simple identification of Omphalotus japonicus using a beam reagent (5 w/v% potassium hydroxide ethanolic solution) were investigated. As an alternative to the beam reagent used for both the direct and extraction methods, a 5 w/v% aqueous solution of sodium sesquicarbonate, a common ingredient used in household detergents, was demonstrated to be capable of identifying Omphalotus japonicus. Furthermore, the concentration of the extract solution of thelephoric acid, which is the coloring molecule, was the same in an 80% ethanol aqueous solution as it was in anhydrous ethanol. This equivalence suggests that ethanol, specified as a disinfectant in the Japanese pharmacopoeia, can be used as an extraction solvent. These findings suggest that identification equivalent to the conventional identification method can be achieved even when using commonly available sodium sesquicarbonate and disinfectant ethanol.

The extensive use of industrial chemicals increases the likelihood of accidental or deliberate environmental release, potentially occurring in occupational, public, or military settings, with skin contact being an important route of exposure. The effectiveness of subsequent skin decontamination depends on the physicochemical properties and absorption profiles of the chemicals involved, as well as the decontamination method applied. This study assessed three solution-based decontamination approaches, hard water, 2% soapy water, and Dahlgren Decon Skin Soap against three representative compounds: chloroacetone, chloroacetonitrile, and crotonaldehyde. For all chemicals tested, statistically significant differences (p < 0.05) were more commonly observed between decontaminated and untreated samples than among the decontamination treatments themselves. These findings indicate that decontamination reliably reduces dermal permeation, even though the specific formulations did not consistently produce distinguishable differences in efficacy. These results support the practical value of prompt decontamination, highlighting that readily accessible solutions can meaningfully mitigate dermal exposure risks in real‑world chemical release scenarios.

Plant-based feedstuffs may have limitations in nutrient digestibility and nutritional value for pigs that might be ameliorated using heat processing technologies. Thus, the main objective was to evaluate the effect of steam-pelleting or extrusion on digestibility of starch, crude protein (CP), amino acids (AA), and gross energy (GE) of the two contrasting ingredients, barley and faba bean in growing pigs. Barley and faba bean ingredients were ground (2.8-mm hammer mill screen), and a portion of each ingredient meal was either steam-pelleted or extruded. Subsequently, the heat-processed ingredients were re-ground. Seven crossbred ileal-cannulated barrows (54.3 ± 3.9 kg) were fed 6 diets containing 95.6% of each of the test ingredients and an N-free diet in a 7 × 7 Latin square. Pigs were fed diets at 2.8 × maintenance digestible energy (DE) during seven 9-d periods consisting sequentially of a 5-d adaptation, 2-d collection of feces, and 2-d collection of digesta. Data were analyzed as a 2 × 3 factorial arrangement. Meal barley contained (as is) 47% starch, 11% CP, 4.8% acid-detergent fiber (ADF), and 2.0% acid-hydrolyzed ether extract (AEE). Meal faba bean contained (as is) 28% starch, 27% CP, 8.6% ADF, and 1.9% AEE. Meal barley had greater (P < 0.05) apparent ileal digestibility (AID) of starch but generally lower (P < 0.05) AID and standardized ileal digestibility (SID) of AA than meal faba bean. Heat processing interacted (P < 0.05) with barley and faba bean on diet AID and SID of CP and most AA, AID of starch, apparent total tract digestibility (ATTD) of GE and starch, and DE value of test ingredients. Steam-pelleting or extruding barley increased (P < 0.05) the ATTD of GE and DE value by 3% but not AID or SID of CP. Steam-pelleting faba bean increased (P < 0.05) the AID of starch, and the AID and SID of CP, Ile, Leu, Met, and Val by 3 to 12%. Extruding faba bean increased (P < 0.05) the AID of starch, and the AID and SID of Ile, Leu, Met, Phe, Thr, and Val by 3% to 10%. Steam-pelleting or extrusion increased (P < 0.05) the predicted NE value of barley and faba bean. In conclusion, heat processing increased the predicted NE value and AA digestibility of faba bean but only the DE and predicted NE value of barley.

Cyclodextrins (CDs) exhibit concentration-dependent aggregation in aqueous solution governed by substitution chemistry, hydration, and intermolecular interactions. In this study, the aggregation behaviour of β-cyclodextrin (β-CD), hydroxypropyl-β-cyclodextrin (HP-β-CD), and sulfobutyl ether-β-cyclodextrin (SBE-β-CD) was systematically investigated in the presence of monovalent (K[+]) and divalent (Ca[2+]) electrolytes using dynamic light scattering, conductivity, surface tension, and phenolphthalein inclusion measurements. Under salt-free conditions, β-CD showed pronounced aggregation driven by intermolecular hydrogen bonding, SBE-β-CD displayed coexistence of molecular and aggregated populations consistent with its charged substituents, and HP-β-CD remained largely dispersed due to steric and hydration effects. Electrolyte addition modified aggregation trends in a valency-dependent manner. K[+] produced moderate changes consistent with electrostatic screening, whereas Ca[2+] induced stronger and time-dependent aggregation effects. Conductivity measurements reflected electrolyte-dependent modification of the ionic environment, while surface tension data supported changes in aggregation state. Phenolphthalein inclusion assays demonstrated that electrolyte-induced aggregation correlates with variations in effective cavity accessibility. These results provide comparative physicochemical insight into how substitution pattern and ion valency influence aggregation and host-guest behaviour of cyclodextrins in aqueous solution.

Cold-water washing has garnered significant attention as an effective strategy for energy conservation and carbon emission reduction. However, most commercially available detergent proteases are derived from mesophilic bacteria, exhibiting limited catalytic efficiency at low temperatures and insufficient source diversity. In this study, a novel cold-adapted subtilisin-like protease, designated PdSP4, was screened and identified from the Antarctic fungus Pseudogymnoascus sp. OUCMDZ-4032. The active band was localized using casein zymography, and the amino acid sequence was elucidated by mass spectrometric analysis, followed by successful heterologous expression in Pichia pastoris and Escherichia coli. Comprehensive enzymatic characterization suggested that PdSP4 retained better catalytic performance toward casein at 0-30 °C than a commercial alkaline protease from Bacillus licheniformis under the tested conditions. The enzyme displays an optimal activity at pH 8.0-9.0, and maintains robust tolerance to various nonionic surfactants and common metal ions. Notably, at 8 °C, PdSP4 effectively removes blood stains from fabrics and displays a pronounced synergistic washing effect when combined with commercial detergents, indicating its strong potential for cold-water laundry applications. In addition, a structure-guided protein engineering strategy led to the construction of a L245G mutant, which further enhanced the low-temperature catalytic activity of PdSP4. Collectively, this study provides a new fungal-derived candidate protease for cold-water detergents and lays a foundation for subsequent protein engineering optimization and industrial application.

Benzalkonium chloride (BAC) and didecyldimethylammonium chloride (DDAC) are quaternary ammonium compound (QAC)-based biocides co-formulated into a wide range of commercial disinfectants, resulting in frequent, simultaneous exposure in humans. Despite their ubiquitous use, the toxicity caused by combined BAC and DDAC exposure remains poorly characterized. In this study, we investigated these effects in human lung adenocarcinoma A549 cells, employing real-time stress granule (SG) formation as a sensitive early-stage biomarker of sub-lethal cell-level stress. Using A549 G3BP1-eGFP knock-in cells, the two compounds induced time- and concentration-dependent SG formation individually and eIF2α phosphorylation, accompanied by a reduction in cell viability and elevation in H₂O₂ levels. Combined exposure at sub-lethal concentrations markedly potentiated SG assembly and synergistically suppressed cell viability beyond levels predicted by the Bliss independence model. At the molecular level, co-exposure robustly amplified activation of multiple unfolded protein response (UPR) branches, including the PERK/eIF2α and IRE1α/XBP1 signaling pathways, whereas individual treatments only partially engaged UPR. Collectively, these findings demonstrate that BAC and DDAC co-exposure synergistically induced toxicity through the cooperative amplification of endoplasmic reticulum stress and oxidative stress signaling, and establish SG formation as a sensitive biomarker for detecting cell-level sub-lethal perturbations not captured by conventional assays. They underscore the need for mixture-based toxicological evaluations of co-formulated disinfectants and highlight the limitations of current frameworks for assessing the safety of single compounds.

Efficient access to soluble recombinant proteins remains a major bottleneck in biochemical and structural studies. We describe an aqueous, solvent-centric, fully miniaturized 96-well workflow to screen extraction conditions that preserve soluble recombinant protein during lysis and clarification in a single working day. Liquid-nitrogen-frozen E. coli pellets are cryogenically bead-milled with stainless-steel beads, retaining the native intracellular milieu while ensuring uniform disruption. The resulting wet, frozen cell powder can therefore be extracted with user-defined solvent, enabling systematic exploration of pH, ionic strength, detergents, and chaotropes. Protein solubility is assessed by a 1 μL chromogenic anti-His dot-blot. We demonstrate the use of the protocol by solubilizing a set of highly challenging de novo-generated proteins and show that dot-blot intensity provides a practical semiquantitative proxy for successful extraction of soluble proteins. We also provide experimentally supported guidelines on the influence of solvent reagents on subsequent steps of protein production, SDS-PAGE, and Ni-NTA purification. This workflow is compatible with upstream genetic solubility-enhancement, chassis- and cultivation-based strategies and enables direct transition from screening hits to scale-up. Because the workflow uses standard molecular biology equipment and inexpensive consumables, it can be readily adopted or automated in most laboratories.