Efficacy of point source legislation quantified for a 64-year river water quality trajectory of phosphorus loading.

Changes in river water quality often follow typical trajectories characterized by sequential phases of degradation and recovery induced by management measures, typically achieved with combinations of legislative and technological interventions. However, the key question about the effectiveness of the different types of legal interventions - source control, use-related, and end-of-pipe - remains poorly understood. With the case of phosphorus (P), which is a valuable indicator of surface water quality and a widespread target of legislation at various governance levels in order to control eutrophication of water bodies, we quantified the relation between point source loading of P and resulting river water quality for a multidecadal trajectory of the river Ruhr (Germany). In particular, we analysed P-related legislation targeting point source pollution enforced at EU, national, state, and local level and linked their development with measured total phosphorus (TP) concentrations in the river Ruhr (Germany). To this end, we combined archival data with information in the literature and conducted interviews with contemporary witnesses to describe and quantify the efficacy of each legislative approach. Although not specifically targeted at P reduction, end-of-pipe measures (sewer systems and wastewater treatment plants (WWTP)) reduced TP inputs to surface waters already in the 1960s and 1970s, curbing TP inputs to the Ruhr by 38% in 1980. The first targeted source control legislation - the banning of phosphates in textile detergents in 1981 - effectively reduced TP concentrations in WWTP influents by 36% since 1990. In combination with stronger end-of-pipe legislation focusing on P elimination in WWTP since the 1990s, TP concentrations in WWTP effluents were reduced by 86% at the end of the 1990s and by 92% in 2021. Complete and successful source control for textile detergents made use-related legislation redundant. Our study demonstrates that source control measures should be prioritized, because they are the fastest way to curb emissions. These findings provide insights that can inform efficient decision-making regarding water quality in a trajectory perspective of hierarchical governance and technological needs, as well as effective policy-making and management for other pollutants requiring control from point sources.

Towards the Development of a Smart Detergent with Enhanced Safety and Efficiency for Pesticide Residue Removal from Fruits and Vegetables.

The research focused on the development and evaluation of special detergents for washing fruits and vegetables, with the primary emphasis on removing pesticide residues. The research aimed to improve food safety and meet consumer preferences for effective cleaning of food products. Using the cloud point characteristic of non-ionic surfactants, a 'smart' detergent was developed to adapt to typical washing conditions. Optimization of the detergent system composition was conducted and the properties of the surfactant system in relation to the cloud point were investigated to highlight the importance of precise control over detergent behavior in response to temperature changes. The physicochemical properties study of the model washing baths included surface tension, aggregate size, solubilization properties, and foaming ability. A model detergent, tailored for both cleaning efficacy and safety against the skin, was developed. Washing efficacy tests demonstrated the superior ability of the designed detergent to remove pesticide residues, eliminating consumer concerns and promoting healthier and safer food consumption. The conducted research paves the way for innovative and safe detergents for washing fruits and vegetables, thereby increasing food safety and consumer satisfaction.

Lipidomics Reveals the Effects of Detergents on Skin Surface Lipids in Young Adults.

BACKGROUND: Detergent is a chemical product commonly used in people's daily life. Contact with detergent solutions can damage the human skin barrier and cause skin diseases. Skin surface lipids (SSLs) play a decisive role in skin barrier function. This study aimed to observe the changes of SSLs in young adults after exposure to detergent solutions to explore the underlying mechanism of skin barrier function damage. METHODS: A self-controlled study on youth adults was conducted in Zhengzhou, China, in November 2020. The study lasted for a total of 1 week, and skin barrier function was assessed by trans-epidermal water loss (TEWL) values. The changes of SSLs before and after exposure to the detergent with subjects were measured using ultra-performance liquid chromatography quadrupole time of flight mass spectrometry. RESULTS: The skin barrier function of subjects' hands was impaired after exposure to detergent (TEWL value increased, p < 0.001). A total of 520 SSLs were detected, divided into 6 main categories. The average relative abundance of these 6 major lipids decreased after exposure. Sphingolipids (mainly ceramides), free fatty acids (mainly long-chain fatty acids), cholesterol lipids, and glycerophospholipids are the most severely damaged lipids. CONCLUSION: Detergent solutions can damage the skin barrier function and SSLs of young hands; interventions targeting SSLs to eliminate detergent damage to human skin may be of value.

Effect of alfalfa supplementary change dietary non-fibrous carbohydrate (NFC) to neutral detergent fiber (NDF) ratio on rumen fermentation and microbial function in Gansu alpine fine wool sheep (Ovis aries).

Bodyweight loss and rumen microbial dysfunction of grazing sheep was a challenge for the sheep production industry during cold season, which were considered to correlated with under-roughage-feeding. Alfalfa is a good roughage supplementary for ruminants, which can improve grazing sheep bodyweight-loss and rumen microbial dysfunction during grass-withering period. This study evaluated the effects of alfalfa hay supplementary change dietary non-fibrous carbohydrate/neutral detergent fiber (NFC/NDF) ratios on rumen fermentation and microbial function of Gansu alpine fine wool sheep during extreme cold season. 120 ewes (3-4 yrs) with an average body weight of 28.71 ± 1.22 kg were allocated randomly into three treatments, and fed NFC/NDF of 1.92 (H group), 1.11 (M group), and 0.68 (L group), respectively. This study was conducted for 107 d, including 7 d of adaption to the diets. The rumen fermentation parameters and microbial characteristics were measured after the end of feeding trials. The results showed that the concentrations of sheep body weight, nitrogen components (Total-N, Soluble protein-N and Ammonia-N), blood biochemical indices (LDH, BUN and CHO) and ruminal volatile fatty acids (TVFA and propionate) significantly increased with an increase in the proportion of NFC/NDF ratios (p < .05), and the acetate and acetate/propionat ratio presented a contrary decreasing trend (p < .05). A total of 1018 OTUs were obtained with 97% consistency. Ruminococcus, Ruminococcaceae and Prevotella were observed as the predominant phyla in ruminal fluid microbiota. Higher NFC/NDF ratios with Alfalfa supplementary increased the richness and diversity of ruminal fluid microbiota, and decreased ruminal fluid microbiota beta-diversity. Using clusters of orthologous groups (COG), the ruminal fluid microbiota of alfalfa supplementary feeding showed low immune pathway and high carbohydrate metabolism pathway. In summary, the study suggested that there was an increasing tendency in dietary NFC/NDF ratio of 1.92 in body weight, ruminal fermentation, microbial community composition and fermentation characteristics through developing alfalfa supplementary system.

Can conventional forages be replaced with cotton plant (Gossypium hirsutum) wastes in fattening lambs? Laboratory and animal studies.

The cotton plant (Gossypium hirsutum) is a shrub native to many arid and semi-arid regions around the world, while the nutritional value of its wastes has been less scientifically investigated. Different components of whole cotton plant wastes (WCPW) including leaf blade, cotton pod, stem, root, bract, petiole, and cottonseed were evaluated for their nutritional values by standard laboratory methods. After that, we tested the WCPW for partial substitution (0, 20, 40, and 60% substitution or 0, 10, 20, and 30% of dietary dry matter (DM)) with dietary common forage in a completely randomized design with 32 feedlot male lambs for 90 days. A diverse range of chemical and mineral compositions was found among the different WCPW's components. The cottonseed had the highest crude protein (CP) and ether extract (EE) contents, while the lowest neutral detergent fiber (NDF) and acid detergent fiber (ADF) were observed in the leaf blade (P < 0.0001). The highest contents of calcium, phosphorus, sodium, magnesium, and iron were also observed in the leaf blade (P < 0.0001). Higher potential gas production, in vitro organic matter digestibility (OMD), in vitro dry matter digestibility (DMD), and total volatile fatty acids (TVFA) were also related to the leaf blade (P < 0.0001). Bract had the highest acid-base buffering capacity (P < 0.0001). The lambs fed on 30% of dietary DM with WCPW exhibited lower final body weight (BW), average daily gain (ADG), CP or NDF digestibility, ruminal TVFA, propionate, plasma total protein, and higher feed conversion ratio (FCR) compared to the control group. Generally, WCPW can be substituted up to 40% of common forages (or 20% of diet DM) without any adverse effect on growth performance and blood metabolites of feedlot lambs, especially during feed shortages.

Evaluation of haemolymph phenoloxidase activity from the grub of Zophobas morio as a predictor of immune response.

In insects, enzyme phenoloxidase plays a critical role in cuticular sclerotisation and defensive functions. In the present investigation, haemolymph phenoloxidase activity from the grub of Zophobas morio was attempted to evaluate as a reliable predictor of insect's immunological response. Among the various substrates tested, L-DOPA was chosen as an appropriate substrate due to its high oxidation. The optimum pH and temperature for haemolymph PO activity was found to be 8 and 30 °C, respectively. The optimum substrate concentration of L-DOPA was found to be 7.5 mM for subsequent PO enzymatic characterisation. Among the various chemical inhibitors and copper chelators, PO activity was significantly reduced in the case of PMSF and thiourea. Preincubation of haemolymph with non-self-molecules showed enhancement of PO activity in the case of LPS from Serratia marcescens. In addition, exogenous proteases like α-chymotrypsin enhanced the PO activity of haemolymph and an increase in PO activity was demonstrated when haemolymph was preincubated with the anionic detergent, SDS and cationic detergent, cetyl pyridium chloride. Alteration of PO activity was observed under agonising conditions of starvation, ligation and microplastics injection at different time intervals. Interestingly, there were no correlation between PO and insect defence under live challenge of microbes. SDS protein profile revealed a significant increase in the 85 kDa and 55 kDa polypeptides in all the experiments over control after 24 h, 48 h and 96 h. Mass spectrophotometric analysis of the polypeptides revealed their homology to antimicrobial peptides for 55 kDa protein and 85 kDa protein. A significant increase in 85 kDa polypeptide was observed in the haemolymph of the grubs after 72 h in the case of starved and microplastics injected groups only. These results demonstrated that PO may not be a reliable benchmark of immunological response in this insect.

Can dietary magnesium sources and buffer change the ruminal microbiota composition and fermentation of lactating dairy cows?

Magnesium oxide (MgO) is one of the most used Mg supplements in livestock. However, to avoid relying upon only one Mg source, it is important to have alternative Mg sources. Therefore, the objective of this study was to evaluate the effects of the interaction of two Mg sources with buffer use on the ruminal microbiota composition, ruminal fermentation, and nutrient digestibility in lactating dairy cows. Twenty lactating Holstein cows were blocked by parity and days in milk into five blocks with four cows each, in a 2 × 2 factorial design. Within blocks, cows were assigned to one of four treatments: 1) MgO; 2) MgO + Na sesquicarbonate (MgO+); 3) calcium-magnesium hydroxide (CaMgOH); 4) CaMgOH + Na sesquicarbonate (CaMgOH+). For 60 d, cows were individually fed a corn silage-based diet, and treatments were top-dressed. Ruminal fluid was collected via an orogastric tube, for analyses of the microbiota composition, volatile fatty acids (VFA), lactate, and ammonia nitrogen (NH3-N). The microbiota composition was analyzed using V4/16S rRNA gene sequencing, and taxonomy was assigned using the Silva database. Statistical analysis was carried out following the procedures of block design analysis, where block and cow were considered random variables. Effects of Mg source, buffer, and the interaction between Mg Source × Buffer were analyzed through orthogonal contrasts. There was no interaction effect of the two factors evaluated. There was a greater concentration of NH3-N, lactate, and butyrate in the ruminal fluid of cows fed with CaMg(OH)2, regardless of the buffer use. The increase in these fermentation intermediates/ end-products can be explained by an increase in abundance of micro-organisms of the genus Prevotella, Lactobacillus, and Butyrivibrio, which are micro-organisms mainly responsible for proteolysis, lactate-production, and butyrate-production in the rumen, respectively. Also, dietary buffer use did not affect the ruminal fermentation metabolites and pH; however, an improvement of the apparent total tract digestibility of dry matter (DM), organic matter (OM), neutral fiber detergent (NDF), and acid fiber detergent (ADF) were found for animals fed with dietary buffer. In summary, there was no interaction effect of buffer use and Mg source, whereas buffer improved total tract apparent digestibility of DM and OM through an increase in NDF and ADF digestibility and CaMg(OH)2 increased ruminal concentration of butyrate and abundance of butyrate-producing bacteria.

Magnesium oxide (MgO) is extensively used as a dietary magnesium (Mg) source in dairy cow diets. However, dairy operations can benefit from other Mg sources. Thus, we evaluated the replacement of dietary MgO with calcium­magnesium hydroxide (CaMg(OH)2) in diets with and without ruminal buffer and their effects on the ruminal microbiota composition, ruminal fermentation, and nutrient digestibility in lactating dairy cows. The study used 20 lactating Holstein cows that were blocked in groups of four and randomly assigned to one of the four treatments. The ruminal content, feed, feces, and urine were collected for analysis of the microbiota composition, ruminal fermentation, nitrogen metabolism, and apparent nutrient digestibility. There was no interaction effect of dietary buffer use and Mg source, while buffer improved total tract apparent digestibility of the dry matter and fiber components; CaMg(OH)2 increased the ruminal concentration of butyrate and the abundance of butyrate-producing bacteria. In summary, we conclude that using CaMg(OH)2 can improve ruminal fermentation regardless of buffer use, which indicates that we can take advantage of the mineral formulation in the diet to modulate the ruminal microbiota composition.

Cyclic pressure induced decellularization of porcine descending aortas.

The demand for decellularized xenogeneic tissues used in reconstructive heart surgery has increased over the last decades. Complete decellularization of longer and tubular aortic sections suitable for clinical application has not been achieved so far. The present study aims at analyzing the effect of pressure application on decellularization efficacy of porcine aortas using a device specifically designed for this purpose. Fresh porcine descending aortas of 8 cm length were decellularized using detergents. To increase decellularization efficacy, detergent treatment was combined with pressure application and different treatment schemes. Quantification of penetration depth as well as histological staining, scanning electron microscopy, and tensile strength tests were used to evaluate tissue structure. In general, application of pressure to aortic tissue does neither increase the decellularization success nor the penetration depth of detergents. However, it is of importance from which side of the aorta the pressure is applied. Application of intermittent pressure from the adventitial side does significantly increase the decellularization degree at the intimal side (compared to the reference group), but had no influence on the penetration depth of SDC/SDS at both sides. Although the present setup does not significantly improve the decellularization success of aortas, it is interesting that the application of pressure from the adventitial side leads to improved decellularization of the intimal side. As no adverse effects on tissue structure nor on mechanical properties were observed, optimization of the present protocol may potentially lead to complete decellularization of larger aortic segments.

The microdomains (rafts) of plasmalemma in the protection of the plant cell under oxidative stress.

The investigation of the lipid-protein microdomains of the plasmalemma isolated with the aid of the non-detergent technique in the zones of the sucrose density gradient after high-speed centrifugation from the tissue pieces of beet roots, which underwent oxidative stress, was conducted. The microdomains, whose lipid composition - according to the definition - allowed us to classify them as rafts, were studied. After the exposure to oxidative stress (100 mM hydrogen peroxide), the variations in the composition of membrane lipids bound up mainly with the elevations of the content of raft-forming lipids (sterols, sterol esters). Oxidative stress provoked redistribution in the composition of sterols, which led to an elevation in the content of campesterol and in the ratio of stigmasterol/sitosterol. Furthermore, the variations were registered in the content of phospholipids and phosphoglycerolipids, which are capable of stabilizing the lamellar structure of membranes. The results obtained allow one to assume that under the oxidative stress, variations in the composition of lipids in microdomains of the plasma membrane can take place. These variations may influence the functioning of the membranes, and the membranes may participate in the protection of the plant cell.

Association of the chemical composition and nutritional value of forage resources in Colombia with methane emissions by enteric fermentation.

In the livestock sector, strategies are available to mitigate gas emissions, such as methane, one of the alternatives that have shown potential correspondence to changes in the composition of the diet. The main aim of this study was to analyze the influence of methane emissions with data on enteric fermentation obtained from the Electronic Data Gathering, Analysis, and Retrieval (EDGAR) database and based on forecasts of methane emissions by enteric fermentation with an autoregressive integrated moving average (ARIMA) model and the application of statistical tests to identify the association between methane emissions from enteric fermentation and the variables of the chemical composition and nutritional value of forage resources in Colombia. The results reported positive correlations between methane emissions and the variables ash content, ethereal extract, neutral detergent fiber (NDF), and acid detergent fiber (ADF) and negative correlations between methane emissions and the variables percentage of unstructured carbohydrates, total digestible nutrients (TDN), digestibility of dry matter, metabolizable energy (MERuminants), net maintenance energy (NEm), net energy gain (NEg), and net lactation energy (NEI). The variables with the most significant influence on the reduction of methane emissions by enteric fermentation are the percentage of unstructured carbohydrates and the percentage of starch. In conclusion, the analysis of variance and the correlations between the chemical composition and the nutritive value of forage resources in Colombia help to understand the influence of diet variables on methane emissions of a particular family and with it in the application of strategies of mitigation.

Supercritical carbon dioxide-decellularized arteries exhibit physiologic-like vessel regeneration following xenotransplantation in rats.

Currently, many techniques are used for decellularization of grafts, including physical, enzymatic, and chemical treatments. Indeed, decellularized xenogenic grafts provide superior outcomes than alternative synthetic conduits. However, vascular grafts produced by these methods are not perfect; their defects include defective vessel wall structures, detergent residues, and the development of aneurysms after grafting. Therefore, it is essential to develop a more appropriate process to produce decellularized vascular grafts. Supercritical carbon dioxide (ScCO2) has been used in decellularization technologies in recent years. It is beneficial for the long-term preservation of tissues and regeneration of new vessels. We have previously reported that ScCO2-produced acellular porcine corneas show excellent biocompatibility following lamellar corneal transplantation in rabbits. In this study, we wanted to use this method to fabricate vascular grafts (ScCO2-decellularized rabbit femoral artery (DFA)) and analyze their efficacy, parameters regarding rejection by the recipient's (ACI/NKyo rats) immune system and biocompatibility, structural regeneration, and functionality in vivo. The results indicated that the ScCO2-DFA showed higher biocompatibility, enhanced chemotactic migration of endothelial progenitor cells, lower risk of vasculopathy, lower inflammatory and splenic immune responses, and better physiological-like tension responses after xenotransplantation (XTP) in ACI/NKyo rats compared with the results obtained after XTP using detergent decellularized vascular grafts (SDS-DFA). In conclusion, ScCO2 is an excellent decellularization technique in the fabrication of biocompatible vascular grafts and has tremendous application in vascular regenerative medicine.

Optimized production of keratinolytic proteases from Bacillus tropicus LS27 and its application as a sustainable alternative for dehairing, destaining and metal recovery.

The present study describes the isolation and characterization of Bacillus tropicus LS27 capable of keratinolytic protease production from Russell Market, Shivajinagar, Bangalore, Karnataka, with its diverse application. The ability of this strain to hydrolyze chicken feathers and skim milk was used to assess its keratinolytic and proteolytic properties. The strain identification was done using biochemical and molecular characterization using the 16S rRNA sequencing method. Further a sequential and systematic optimization of the factors affecting the keratinase production was done by initially sorting out the most influential factors (NaCl concentration, pH, inoculum level and incubation period in this study) through one factor at a time approach followed by central composite design based response surface methodology to enhance the keratinase production. Under optimized levels of NaCl (0.55 g/L), pH (7.35), inoculum level (5%) and incubation period (84 h), the keratinase production was enhanced from 41.62 U/mL to 401.67 ± 9.23 U/mL (9.65 fold increase) that corresponds to a feather degradation of 32.67 ± 1.36% was achieved. With regard to the cost effectiveness of application studies, the crude enzyme extracted from the optimized medium was tested for its potential dehairing, destaining and metal recovery properties. Complete dehairing was achieved within 48 h of treatment with crude enzyme without any visible damage to the collagen layer of goat skin. In destaining studies, combination of crude enzyme and detergent solution [1 mL detergent solution (5 mg/mL) and 1 mL crude enzyme] was found to be most effective in removing blood stains from cotton cloth. Silver recovery from used X-ray films was achieved within 6 min of treatment with crude enzyme maintained at 40 °C.

Contribution of household dishwashing to microplastic pollution.

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.

Ionic Detergent Under Pressure-Vacuum as an Innovative Strategy to Generate Canine Tracheal Scaffold for Organ Engineering.

Decellularized scaffolds applied in tissue engineering offer improvements, supplying the elevated necessity for organs and tissues for replacement. However, obtaining a functional trachea for autotransplantation or allotransplantation is tricky due to the organ anatomical and structural complexity. Most tracheal decellularization protocols are lengthy, expensive, and could damage the tracheal extracellular matrix (ECM) architecture and functionality. Here, we aimed to evaluate the effectiveness of 3 different decellularization protocols combined with chemical and physical methods to obtain acellular canine tracheal scaffolds. Six adult dog tracheas were incised (tracheal segments) resulting in 28 rings for control tissue and 84 rings for decellularization (5-7 mm thick). Subsequently, decellularized tracheal scaffolds were microscopically/macroscopically characterized by histological analysis (Hematoxylin-Eosin, Masson's trichrome, Picrosirius red, Alcian blue, and Safranin O), immunohistochemistry for ECM components, scanning electron microscopy, and genomic DNA quantification. After decellularization, the tracheal tissue revealed reduced genomic DNA, and maintenance of ECM components preserved (structural proteins, adhesive glycoproteins, glycosaminoglycans and proteoglycans), suggesting ECM integrity and functionality. Comparatively, the combined ionic detergent with high vacuum pressure decellularization protocol revealed superior genomic DNA decrease (13.5 ng/mg) and improvement on glycosaminoglycans and proteoglycans preservation regarding the other decellularized trachea scaffolds and native tissue. Our results indicate that the 3 chemical/physical protocols reduce the decellularization time without ECM proteins damage. Notwithstanding, the use of ionic detergent under vacuum pressure was able to generate an innovative strategy to obtain acellular canine tracheal scaffolds with the highest levels of adhesive proteins that support its potentiality for recellularization and future tissue engineering application.

Heterogeneous chemistry of ozone with floor cleaning agent: Implications of secondary VOCs in the indoor environment.

Human daily activities such as cooking, and cleaning can affect the indoor air quality by releasing primary emitted volatile organic compounds (VOCs), as well as by the secondary product compounds formed through reactions with ozone (O3) and hydroxyl radicals (OH). However, our knowledge about the formation processes of the secondary VOCs is still incomplete. We performed real-time measurements of primary VOCs released by commercial floor-cleaning detergent and the secondary product compounds formed by heterogeneous reaction of O3 with the constituents of the cleaning agent by use of high-resolution mass spectrometry. We measured the uptake coefficients of O3 on the cleaning detergent at different relative humidities in dark and under different light intensities (320 nm < λ < 400 nm) relevant for the indoor environment. On the basis of the detected compounds we developed tentative reaction mechanisms describing the formation of the secondary VOCs. Intriguingly, under light irradiation the formation of valeraldehyde was observed based on the photosensitized chemistry of acetophenone which is a constituent of the cleaning agent. Finally, we modeled the observed mixing ratios of three aldehydes, glyoxal, methylglyoxal, and 4-oxopentanal with respect to real-life indoor environment. The results suggest that secondary VOCs initiated by ozone chemistry can additionally impact the indoor air pollution.

Miniprep assisted proteomics (MAP) for rapid proteomics sample preparation.

Complete enzymatic digestion of proteins for bottom-up proteomics is substantially improved by use of detergents for denaturation and solubilization. Detergents however, are incompatible with many proteases and highly detrimental to LC-MS/MS. Recently; filter-based methods have seen wide use due to their capacity to remove detergents and harmful reagents prior to digestion and mass spectrometric analysis. We hypothesized that non-specific protein binding to negatively charged silica-based filters would be enhanced by addition of lyotropic salts, similar to DNA purification. We sought to exploit these interactions and investigate if low-cost DNA purification spin-filters, 'Minipreps,' efficiently and reproducibly bind proteins for digestion and LC-MS/MS analysis. We propose a new method, Miniprep Assisted Proteomics (MAP), for sample preparation. We demonstrate binding capacity, performance, recovery and identification rates for proteins and whole-cell lysates using MAP. MAP recovered equivalent or greater protein yields from 0.5-50 µg analyses benchmarked against commercial trapping preparations. Nano UHPLC-MS/MS proteome profiling of lysates of Escherichia coli had 99.3% overlap vs. existing approaches and reproducibility of replicate minipreps was 98.8% at the 1% FDR protein level. Label Free Quantitative proteomics was performed and 91.2% of quantified proteins had a %CV <20% (2044/2241). Miniprep Assisted Proteomics can be performed in minutes, shows low variability, high recovery and proteome depth. This suggests a significant role for adventitious binding in developing new proteomics sample preparation techniques. MAP represents an efficient, ultra-low-cost alternative for sample preparation in a commercially obtainable device that costs ∼$0.50 (USD) per miniprep.

Facile and scalable fabrication of exosome-mimicking nanovesicles through PEGylated lipid detergent-aided cell extrusion.

We report a scalable fabrication method to generate exosome-mimicking nanovesicles (ENVs) by using a biocompatible, cell-binding lipid detergent during cell extrusion. A PEGylated mannosylerythritol lipid (MELPEG) detergent was rationally engineered to strongly associate with phospholipid membranes to increase cell membrane deformability and the corresponding friction force during extrusion and to enhance the dispersibility of ENVs. Compared to cell extrusion without detergent, cell extrusion in the presence of MELPEG increased the ENV production yield by approximately 20 times and cellular protein content per MELPEG-functionalized ENV by approximately 2-fold relative to that of unmodified ENVs. We verified that MELPEG strongly binds to ENV membranes and increases membrane deformability via expansion/swelling while preserving the integrity of the phospholipid bilayer structure. The results highlight that the MELPEG-aided cell extrusion process broadly applies to various cell lines; hence, it could be helpful in the production of ENVs for tissue regeneration, drug delivery, and cancer nanomedicine.

Prediction of methane per unit of dry matter intake in growing and finishing cattle from the ratio of dietary concentrations of starch to neutral detergent fiber alone or in combination with dietary concentration of ether extract.

Previous research demonstrated that a fixed value of 0.2433 (SE = 0.0134) Mcal of CH4/kg of dry matter intake (DMI) could be used to predict CH4 production with accuracy and precision on par with similar equations in the literature. Slope bias was substantially less for the fixed-coefficient equation than noted for the other DMI- or gross energy intake (GEI)-based equations, but mean bias was substantially greater, presumably reflecting the failure of the fixed-coefficient approach to account for dietary factors that affect CH4 production. In this article, we report on the use of the dietary ratio of concentrations of starch to neutral detergent fiber (NDF) and dietary ether extract (EE) concentration to improve the accuracy and precision of the fixed-coefficient equation. The same development data set used to create the fixed-coefficient equation was used in the present study, which included 134 treatment means from 34 respiration calorimetry studies. Based on stepwise regression with dietary NDF, starch, crude protein, EE, and the starch:NDF ratio as possible dependent variables, the starch:NDF ratio and EE were the only dietary variables selected (P ≤ 0.15). The study-adjusted relationship with the starch:NDF ratio (r2 = 0.673; root mean square error [RMSE] = 0.0327) was: Mcal of CH4/kg of DMI = 0.2883 - 0.03474 × starch:NDF; whereas the relationship with a model that included both starch:NDF ratio and dietary EE (r2 = 0.738; RMSE = 0.0315) was: Mcal of CH4/kg of DMI = 0.3227 - 0.0334 × starch:NDF - 0.00868 × % EE. A previously published independent data set with 129 treatment means from 30 respiration calorimetry studies was used to evaluate these two equations, along with two additional equations in which g/d of CH4 was predicted directly from DMI, starch:NDF ratio, and/or dietary EE. The two Mcal of CH4/kg of DMI equations had superior fit statistics to the previously published 0.2433 Mcal of CH4/kg of DMI equation, with a substantial decrease in mean bias and improved concordance correlation coefficients. Moreover, the Mcal of CH4/kg of DMI equations resulted in improved fit relative to direct prediction of g/d of CH4 from DMI, the starch:NDF ratio, and % EE. Based on these results, further evaluation of the dietary ratio of starch-to-NDF concentrations and EE concentration to predict methane production per unit DMI in beef cattle is warranted.

Accurate and precise equations to predict enteric methane production are an important tool as the beef industry works to mitigate greenhouse gas emissions. We predicted the megacalories of methane per unit of dry matter intake using data from growing and finishing beef and dairy steers and heifers. The predicted value was then multiplied by dry matter intake to estimate methane production. The most important predictor variables were the ratio of dietary starch to neutral detergent fiber concentrations and dietary ether extract concentration. When tested with an independent data set, predicting methane per unit dry matter intake from the ratio of starch to neutral detergent fiber, with or without the addition of dietary ether extract as a predictor variable, yielded better fits to the independent data set than equations in which the daily production of methane was predicted directly from dry matter intake, the ratio of starch to neutral detergent fiber, and/or dietary ether extract. Predictions could potentially be further refined by adjusting the starch component of the ratio for degradability in the rumen (e.g., degree of gelatinization or processing) and the neutral detergent fiber content for the contributions from roughage vs. grains and grain byproducts.

Effect of coagulation on microfibers in laundry wastewater.

Microplastics pollution in the aquatic system has received significant attention due to their recalcitrant nature and ecotoxicological threat. Municipal wastewater typically contains various microplastics with synthetic microfibers as a significant constituent from the laundry process. The fate of microfibers in conventional wastewater processes is not clearly understood. In this study, the effect of coagulation on microfibers obtained from a lint screen of a domestic dryer and resuspended in pure water, and also in laundry wastewater was investigated using ferric chloride and polyaluminum chloride (PACl). The removal efficiency of the microfibers resuspended in pure water varied from 86% to 96% depending on the fiber size ranges: < 90 µm, 90-125 µm, and >125 µm with the smaller size microfibers showing a lower removal efficiency. Surfactant present in detergent in laundry wastewater reduced the microfibers removal efficiency to 0-37%, however, the addition of PACl increased microfibers removal to 90%. The optimal PACl concentrations for ≥90% removal were 1.75, 2, 4, and 6 mg/L for 0.5, 2, 4, 8 mg/L detergent, respectively. Zeta potential, FTIR, and SEM analysis were applied to observe the surface changes of microfibers during coagulation indicating possible mechanisms of coagulation. The dominant mechanisms for coagulation of microfibers by FeCl3 and PACl seem to be charge neutralization and adsorption-bridging. This work provided some insights about the fate of laundry microfibers in primary treatment processes.

Aponeurosis discission, a low-detergent method for tissue-engineered acellular ligament scaffolds.

Detergent treatment is the most commonly used method for the decellularization of ligaments and tendon grafts. However, it is well recognized that detergent treatment can also adversely affect the extracellular matrix. This study found that discission into the aponeurosis layer of the patellar tendon (PT) before decellularization is conducive to extracting cells from the PT using a low quantity of detergent in a short time period. The acellular aponeurosis discission ligament (AADL) retains its native collagen fibril structure and mechanical properties. Moreover, the PT retained cell and tissue compatibility in vitro and in vivo. After implantation into a defective allogeneic PT, we found that the AADL healed well in the host, and its collagen structure exhibited gradual improvement 12 months after implantation with satisfactory reconstruction. IMPACT: The aponeurosis of tendons/ligaments is the main barrier to achieving complete decellularization, and it thus prevents complete recellularization for applications in tissue engineering. Aponeurosis can obstruct the removal of cell components. We found that excising the aponeurosis before decellularization allows for the removal of cellular components with a reduced amount of detergent, thus improving the biological properties of the acellular ligament. To the best of our knowledge, no similar studies have been performed. Graphical abstract.