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1.
Protein Sci ; 33(6): e4976, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38757374

RESUMO

G-protein coupled receptors (GPCRs) are the largest class of membrane proteins encoded in the human genome with high pharmaceutical relevance and implications to human health. These receptors share a prevalent architecture of seven transmembrane helices followed by an intracellular, amphipathic helix 8 (H8) and a disordered C-terminal tail (Ctail). Technological advancements have led to over 1000 receptor structures in the last two decades, yet frequently H8 and the Ctail are conformationally heterogeneous or altogether absent. Here we synthesize a peptide comprising the neurotensin receptor 1 (NTS1) H8 and Ctail (H8-Ctail) to investigate its structural stability, conformational dynamics, and orientation in the presence of detergent and phospholipid micelles, which mimic the membrane. Circular dichroism (CD) and nuclear magnetic resonance (NMR) measurements confirm that zwitterionic 1,2-diheptanoyl-sn-glycero-3-phosphocholine is a potent stabilizer of H8 structure, whereas the commonly-used branched detergent lauryl maltose neopentyl glycol (LMNG) is unable to completely stabilize the helix - even at amounts four orders of magnitude greater than its critical micellar concentration. We then used NMR spectroscopy to assign the backbone chemical shifts. A series of temperature and lipid titrations were used to define the H8 boundaries as F376-R392 from chemical shift perturbations, changes in resonance intensity, and chemical-shift-derived phi/psi angles. Finally, the H8 azimuthal and tilt angles, defining the helix orientation relative of the membrane normal were measured using paramagnetic relaxation enhancement NMR. Taken together, our studies reveal the H8-Ctail region is sensitive to membrane physicochemical properties and is capable of more adaptive behavior than previously suggested by static structural techniques.


Assuntos
Receptores de Neurotensina , Receptores de Neurotensina/química , Receptores de Neurotensina/metabolismo , Receptores de Neurotensina/genética , Humanos , Micelas , Ressonância Magnética Nuclear Biomolecular , Peptídeos/química , Peptídeos/metabolismo , Dicroísmo Circular , Conformação Proteica em alfa-Hélice , Detergentes/química , Modelos Moleculares
2.
Sci Rep ; 14(1): 10888, 2024 05 13.
Artigo em Inglês | MEDLINE | ID: mdl-38740835

RESUMO

Ethylenediaminetetraacetic acid (EDTA), a classically used chelating agent of decalcification, maintains good morphological details, but its slow decalcification limits its wider applications. Many procedures have been reported to accelerate EDTA-based decalcification, involving temperature, concentration, sonication, agitation, vacuum, microwave, or combination. However, these procedures, concentrating on purely tissue-outside physical factors to increase the chemical diffusion, do not enable EDTA to exert its full capacity due to tissue intrinsic chemical resistances around the diffusion passage. The resistances, such as tissue inner lipids and electric charges, impede the penetration of EDTA. We hypothesized that delipidation and shielding electric charges would accelerate EDTA-based penetration and the subsequent decalcification. The hypothesis was verified by the observation of speedy penetration of EDTA with additives of detergents and hypertonic saline, testing on tissue-mimicking gels of collagen and adult mouse bones. Using a 26% EDTA mixture with the additives at 45°C, a conventional 7-day decalcification of adult mouse ankle joints could be completed within 24 h while the tissue morphological structure, antigenicity, enzymes, and DNA were well preserved, and mRNA better retained compared to using 15% EDTA at room temperature. The addition of hypertonic saline and detergents to EDTA decalcification is a simple, rapid, and inexpensive method that doesn't disrupt the current histological workflow. This method is equally or even more effective than the currently most used decalcification methods in preserving the morphological details of tissues. It can be highly beneficial for the related community.


Assuntos
Detergentes , Ácido Edético , RNA Mensageiro , Animais , Ácido Edético/química , Ácido Edético/farmacologia , Detergentes/química , Camundongos , RNA Mensageiro/genética , Solução Salina Hipertônica/química , Osso e Ossos/metabolismo , Osso e Ossos/efeitos dos fármacos , Osso e Ossos/química , Técnica de Descalcificação/métodos
3.
Trop Anim Health Prod ; 56(4): 126, 2024 Apr 16.
Artigo em Inglês | MEDLINE | ID: mdl-38625501

RESUMO

The objective of this study was to compare the indirect methods of obtaining digestibility with the direct method of total fecal collection to estimate the apparent digestibility of nutrients in sheep and goats supplemented with non-protein nitrogen. Five goats and five sheep with no defined racial pattern were used, distributed in two 5 × 5 Latin squares, with split plots, considering the diets as plots and the apparent digestibility determination methodologies as subplots. The diets were composed of buffelgrass hay and the addition, via ruminal infusion, of increasing amounts of nitrogen supplementation in order to gradually raise the CP level of the basal diet in intervals of 2% points, that is, + 2, +4, + 6 and + 8%. Samples of the feeds offered, and the leftovers were collected daily during the five days of collection to determine the nutrient intake, as well as the total collection of feces to determine the apparent digestibility of the nutrients. The amount of fecal dry matter excreted was estimated by the concentration of Indigestible Acid Detergent Fiber (ADFi), Indigestible Neutral Detergent Fiber (NDFi), Indigestible Dry Matter at 244 h (DMi 244 h) and Indigestible Dry Matter at 264 h (DMi 264 h). Among the evaluated markers, DMi 264 h had the lowest accuracy in estimating fecal excretion and nutrient digestibility. For the goat species, the markers ADFi and DMi 244 h proved to be able to adequately predict fecal excretion and digestibility indices, while NDFi stood out for both species. Among the evaluated markers, NDFi is the one that most accurately estimates the nutrient digestibility of the diet for goats and sheep.


Assuntos
Detergentes , Cabras , Animais , Ovinos , Nutrientes , Suplementos Nutricionais , Nitrogênio
4.
Microb Cell Fact ; 23(1): 109, 2024 Apr 12.
Artigo em Inglês | MEDLINE | ID: mdl-38609920

RESUMO

BACKGROUND: Cellulase is considered a group member of the hydrolytic enzymes, responsible for catalyzing the hydrolysis of cellulose and has various industrial applications. Agricultural wastes are used as an inexpensive source for several utilizable products throughout the world. So, searching for cellulase enzymes from fungal strains capable of utilizing agricultural wastes to increase productivity, reduce costs and overcome waste accumulation in the environment is very important to evaluate its potency as a bio-additive to detergent agents. RESULTS: In the current study, the previously identified fungal strain Aspergillus terreus MN901491 was screened and selected for cellulase production. Medium parameters were optimized using one-factor-at-a-time (OFAT) and multi-factorial (Plackett-Burman and Box-Behnken) design methods. OFAT showed the ability of the fungal strain to utilize agricultural wastes (corn cob and rice straw) as a substrate. Also, yeast extract was the best nitrogen source for enhancing cellulase productivity. The most significant variables were determined by Plackett-Burman Design (PBD) and their concentrations were optimized by Response Surface Methodology (RSM) using Box-Behnken Design (BBD). Among eleven independent variables screened by PBD, malt extract, (NH4)2SO4, and KCl were the most significant ones followed by rice straw which affected cellulase production positively. The ANOVA results particularly the R2-value of PBD (0.9879) and BBD (0.9883) confirmed the model efficiency and provided a good interpretation of the experiments. PBD and BBD improved cellulase productivity by 6.1-fold greater than that obtained from OFAT. Medium optimization using OFAT and statistical models increased cellulase production from A. terreus MN901491 by 9.3-fold compared to the non-optimized medium. Moreover, the efficiency of cellulase activity on cotton fabrics as a bio-additive detergent was evaluated and estimated using whiteness and scanning electron microscope (SEM) that affirmed its potential effect and remarkable detergent ability to improve whiteness by 200% in comparison with non-washed fabric and by 190% in comparison with fabric washed by water. CONCLUSION: The presented work was stabilized as a multi-efficiency in which wastes were used to produce cellulase enzyme from the fungal strain, Aspergillus terreus MN901491 as a bio-additive to detergent applications that involved ecofriendly and green processes.


Assuntos
Celulase , Oryza , Detergentes , Aspergillus , Projetos de Pesquisa
5.
PLoS One ; 19(4): e0300864, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38635849

RESUMO

Chia (Salvia hispanica L.) seed (CS) and Pumpkin (Cucurbita moschata) seed (PS) are used in ruminant diets as energy sources. The current experiment studied the impact of dietary inclusion of CS and PS on nutrient intake and digestibility, milk yield, and milk composition of dairy sheep. Twelve primiparous Texel × Suffolk ewes [70 ± 5 days in milk (DIM); 0.320 ± 0.029 kg milk yield] were distributed in a 4 × 3 Latin square design and fed either a butter-based control diet [CON; 13 g/kg dry matter] or two diets with 61 g/kg DM of either CS or PS. Dietary inclusion of CS and PS did not alter live weight (p >0.1) and DM intake (p >0.1). However, compared to the CON, dietary inclusion of both CS and PS increased the digestibility of neutral detergent fiber (p <0.001) and acid detergent lignin (p < 0.001). Milk production (p = 0.001), fat-corrected milk (p < 0.001), and feed efficiency (p < 0.001) were enhanced with PS, while the highest milk protein yield (p < 0.05) and lactose yield (p < 0.001) were for CS-fed ewes. Compared to the CON diet, the ingestion of either CS and/or PS decreased (p < 0.001) the C16:0 in milk. Moreover, both CS and PS tended to enhance the content of C18:3n6 (p > 0.05) and C18:3n3 (p > 0.05). Overall short-term feeding of CS and/or PS (up to 6.1% DM of diet) not only maintains the production performance and digestibility of nutrients but also positively modifies the milk FA composition.


Assuntos
Cucurbita , Animais , Feminino , Ovinos , Cucurbita/metabolismo , Lactação , Salvia hispanica , Detergentes , Fibras na Dieta/metabolismo , Dieta/veterinária , Sementes/metabolismo , Digestão , Ração Animal/análise , Zea mays/metabolismo , Suplementos Nutricionais/análise , Rúmen/metabolismo
6.
J Anim Sci ; 1022024 Jan 03.
Artigo em Inglês | MEDLINE | ID: mdl-38581217

RESUMO

Pelleted total mixed ration (P-TMR) feeding, which has become a common practice in providing nutrition for fattening sheep, requires careful consideration of the balance between forage neutral detergent fiber (FNDF) and rumen degradable starch (RDS) to maintain proper rumen functions. The present study aimed to investigate the effects of the dietary FNDF/RDS ratio (FRR) on chewing activity, ruminal fermentation, ruminal microbes, and nutrient digestibility in Hu sheep fed a P-TMR diet. This study utilized eight ruminally cannulated male Hu sheep, following a 4 × 4 Latin square design with 31 d each period. Diets consisted of four FRR levels: 1.0 (high FNDF/RDS ratio, HFRR), 0.8 (middle high FNDF/RDS ratio, MHFRR), 0.6 (middle low FNDF/RDS ratio, MLFRR), and 0.4 (low FNDF/RDS ratio, LFRR). Reducing the dietary FRR levels resulted in a linear decrease in ruminal minimum pH and mean pH, while linearly increasing the duration and area of pH below 5.8 and 5.6, as well as the acidosis index. Sheep in the HFRR and MHFRR groups did not experience subacute ruminal acidosis (SARA), whereas sheep in another two groups did. The concentration of total volatile fatty acid and the molar ratios of propionate and valerate, as well as the concentrate of lactate in the rumen linearly increased with reducing dietary FRR, while the molar ratio of acetate and acetate to propionate ratio linearly decreased. The degradability of NDF and ADF for alfalfa hay has a quadratic response with reducing the dietary FRR. The apparent digestibility of dry matter, organic matter, neutral detergent fiber, and acid detergent fiber linearly decreased when the dietary FRR was reduced. In addition, reducing the dietary FRR caused a linear decrease in OTUs, Chao1, and Ace index of ruminal microflora. Reducing FRR in the diet increased the percentage of reads assigned as Firmicutes, but it decreased the percentage of reads assigned as Bacteroidetes in the rumen. At genus level, the percentage of reads assigned as Prevotella, Ruminococcus, Succinivibrio, and Butyrivibrio linearly decreased when the dietary FRR was reduced. The results of this study demonstrate that the dietary FRR of 0.8 is crucial in preventing the onset of SARA and promotes an enhanced richness of ruminal microbes and also improves fiber digestibility, which is a recommended dietary FRR reference when formulating P-TMR diets for sheep.


Forage neutral detergent fiber (FNDF) and rumen degradable starch (RDS) are key components of carbohydrates in the diet for ruminants, which would reflect saliva secretion and the acid production potential of feed. However, appropriate FNDF to RDS ratios (FRR) applicable to ruminants under the condition of pelleted total mixed ration (P-TMR) feeding have not been reported. In this study, we investigated the effects of the dietary FRR on chewing activity, ruminal fermentation, ruminal microbial communities, and nutrient digestibility of Hu sheep under P-TMR feeding. The results indicate that reducing dietary FRR levels would induce acidosis in sheep, which negatively affected fiber utilization and ruminal bacterial communities. The FRR of 0.8 was a recommended dietary FRR when formulating a P-TMR diet for fattening sheep, as indicated by decreased ruminal acidosis risk and increased richness of ruminal microbes in the rumen as well as nutrient digestibility.


Assuntos
Acidose , Doenças dos Ovinos , Masculino , Feminino , Animais , Ovinos , Leite/metabolismo , Mastigação/fisiologia , Amido/metabolismo , Lactação/fisiologia , Detergentes/metabolismo , Silagem/análise , Propionatos/metabolismo , Fermentação , Rúmen/metabolismo , Fibras na Dieta/metabolismo , Carboidratos da Dieta/metabolismo , Dieta/veterinária , Nutrientes , Acetatos/metabolismo , Acidose/veterinária , Digestão/fisiologia
7.
Environ Sci Pollut Res Int ; 31(21): 30497-30508, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38607492

RESUMO

Detergents are highly produced pollutants with environmental problems like foam generation and toxic effects in biota. Nonylphenol ethoxylates (NPEs) are efficient, economical, and versatile surfactants, used in detergents for more than 40 years due to their detergency capacity. In the environment, NPE biodegrades into the metabolite nonylphenol (NP), classified as an endocrine disruptor. The identification and quantification of 4-NP in a designed detergent and 30 commercially available detergents were performed to prove the degradation of NPE into 4-NP during storage time. This investigation introduces the first evidence of NPE degradation during storage in commercially available detergents, demonstrating a novel exposure pathway in humans that has not been explored before, representing potential human health risks. Therefore, simple, easy, low-cost, and available approaches to remove and substitute NP is paramount. Alkyl polyglucoside (APG) was assessed as a substitute, and the feasibility of this substitution was proven according to physical and chemical properties, cleaning performance, and antimicrobial properties. NPE substitution in detergents is demonstrated as a viable strategy to minimize exposure risks in humans and the environment.


Assuntos
Detergentes , Detergentes/química , Etilenoglicóis/química , Fenóis , Tensoativos/química , Humanos , Disruptores Endócrinos/análise
8.
Arch Med Sadowej Kryminol ; 73(3): 272-277, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38662468

RESUMO

Toilet cleaner containing hydrochloric acid is a common item found in households all over the world. Due to the availability of the substance, it becomes one of the main contributors to corrosive damage to the gastrointestinal system. This study reports a case of a female in her 50s with an alleged history of ingestion of toilet cleaner an empty bottle of which was found together with a suicide note at the incident site. During the autopsy, the forensic expert made an intriguing observation regarding the dispersion of ingested acid to other organs without gastric perforation. Despite the absence of gastric perforation, the corrosive effects of the ingested acid were evident in various organs, including the liver and spleen. This phenomenon suggests a unique mechanism by which the acid is able to disperse and cause damage beyond the stomach, leading to widespread organ involvement. However, through a comprehensive analysis of the detailed history, typical macroscopic autopsy findings, and chemical analysis reports, it is possible to establish that the cause of death is corrosive acid poisoning. In such cases, further investigation is warranted to gain a better understanding of the underlying mechanisms responsible for the dispersion of the acid and its clinical implications. By delving deeper into these aspects, we can enhance our knowledge and contribute to the field of forensic medicine.


Assuntos
Autopsia , Humanos , Feminino , Pessoa de Meia-Idade , Ácido Clorídrico/efeitos adversos , Ácido Clorídrico/intoxicação , Cáusticos/intoxicação , Cáusticos/toxicidade , Queimaduras Químicas/patologia , Queimaduras Químicas/etiologia , Detergentes/intoxicação , Detergentes/efeitos adversos , Suicídio Consumado , Patologia Legal
9.
PLoS One ; 19(4): e0284346, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38648212

RESUMO

Child impeding packs are difficult for children to open so protect them from unintended access to hazardous contents inside the pack. However, if packs are difficult for adults to open, in normal usage scenarios, this may result in a higher occurrence of packs being left open. This research explores differences in ease of usage and closure compliance between two types of child impeding packs of liquid laundry detergent capsules. The two packs, "Pinch & Lift" and "Press & Lift", had different opening and closing mechanisms. "Press & Lift" also included an audible "click" signal to confirm complete closure to the user. The research was performed across two studies. In both studies, the packs were used in participants' homes according to their usual storage and usage practices as replacements to their current liquid laundry detergent capsule pack. All participants had small children living with them in their household. In Study 1, self-reported closure and ease of use data was collected from 99 adult participants in Germany. They used each package in their home for 10 days. Study 2 extended Study 1 by measuring closure rates with an objective assessment using in pack sensors for a 10-day period for each pack. Self-reported closure and ease of use data were also collected. Study 2 was conducted with 87 participants in the United Kingdom. Results across both studies showed "Press & Lift" with the audible "Click" close signal to be rated by participants to be significantly easier to open and close and have a higher self-reported closure rate than "Pinch & Lift". In addition, Study 2 results demonstrated higher closure rates using "Press & Lift" based on the sensor-measured closure compliance. Together, the results suggest transition to a pack with a mechanism that is child impeding and easier for an adult to use with an audible closure signal, like the "Press & Lift" system, has potential to reduce child access to a capsule from a pack by reducing the likelihood of the pack being left open by the adult user. Ultimately, such packs could protect children from potential poisoning injury across a range of consumer products.


Assuntos
Detergentes , Humanos , Adulto , Feminino , Masculino , Criança , Embalagem de Produtos/métodos , Pessoa de Meia-Idade , Adulto Jovem , Reino Unido , Alemanha , Qualidade de Produtos para o Consumidor
10.
Int J Mol Sci ; 25(8)2024 Apr 20.
Artigo em Inglês | MEDLINE | ID: mdl-38674110

RESUMO

Membrane proteins constitute about 20% of the human proteome and play crucial roles in cellular functions. However, a complete understanding of their structure and function is limited by their hydrophobic nature, which poses significant challenges in purification and stabilization. Detergents, essential in the isolation process, risk destabilizing or altering the proteins' native conformations, thus affecting stability and functionality. This study leverages single-particle cryo-electron microscopy to elucidate the structural nuances of membrane proteins, focusing on the SLAC1 bacterial homolog from Haemophilus influenzae (HiTehA) purified with diverse detergents, including n-dodecyl ß-D-maltopyranoside (DDM), glycodiosgenin (GDN), ß-D-octyl-glucoside (OG), and lauryl maltose neopentyl glycol (LMNG). This research not only contributes to the understanding of membrane protein structures but also addresses detergent effects on protein purification. By showcasing that the overall structural integrity of the channel is preserved, our study underscores the intricate interplay between proteins and detergents, offering insightful implications for drug design and membrane biology.


Assuntos
Proteínas de Bactérias , Microscopia Crioeletrônica , Detergentes , Haemophilus influenzae , Microscopia Crioeletrônica/métodos , Haemophilus influenzae/ultraestrutura , Haemophilus influenzae/química , Proteínas de Bactérias/química , Proteínas de Bactérias/ultraestrutura , Detergentes/química , Microscopia Eletrônica de Transmissão/métodos , Proteínas de Membrana/química , Proteínas de Membrana/ultraestrutura , Proteínas de Membrana/metabolismo
11.
Protein Expr Purif ; 219: 106479, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38574878

RESUMO

Owing to vast therapeutic, commercial, and industrial applications of microbial proteases microorganisms from different sources are being explored. In this regard, the gut microbiota of Monopteruscuchia were isolated and examined for the production of protease. All the isolates were primarily and secondarily screened on skim milk and gelatin agar plates. The protease-positive isolates were characterized morphologically, biochemically, and molecularly. Out of the 20 isolated strains,6 belonging to five different genera viz.Bacillus,Priestia,Aeromonas,Staphylococcus, and Serratia demonstrated proteolytic activity. Bacillussafensis strain PRN1 demonstrated the highest protease production and, thus, the largest hydrolytic clear zones in both skim milk agar (15 ± 1 mm) and gelatin (16 ± 1 mm) plates. The optimized parameters (time, pH, temperature, carbon, nitrogen) for highest protease activity and microbial growth of B.safensis strain PRN1 includes 72 h (OD600 = 0.56,1303 U/mL), pH 8 (OD600 = 0.83, 403.29 U/mL), 40 °C (OD600 = 1.75, 1849.11 U/mL), fructose (OD600 = 1.22, 1502 U/mL), and gelatin (OD600 = 1.88, 1015.33 U/mL). The enzyme was purified to homogeneity using salt-precipitation and gel filtration chromatography. The sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) demonstrated that the purified enzyme was a monomer of a molecular weight of ∼33 kDa. The protease demonstrated optimal activity at pH 8 and 60 °C. It was strongly inhibited by phenylmethylsulfonyl fluoride (PMSF), demonstrating that it belongs to the serine-proteases family. The compatibility of the enzyme with surfactants and commercial detergents demonstrates its potential use in the detergent industry. Furthermore, the purified enzyme showed antibacterial and blood-stain removal properties.


Assuntos
Bacillus , Detergentes , Serina Proteases , Detergentes/química , Detergentes/farmacologia , Serina Proteases/isolamento & purificação , Serina Proteases/química , Serina Proteases/genética , Serina Proteases/metabolismo , Bacillus/enzimologia , Bacillus/genética , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Proteínas de Bactérias/isolamento & purificação , Proteínas de Bactérias/metabolismo , Concentração de Íons de Hidrogênio
12.
PDA J Pharm Sci Technol ; 78(2): 187-195, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38609155

RESUMO

The use of detergents or low pH hold are commonly employed techniques in biologics downstream processing to inactivate enveloped viruses. These approaches have been demonstrated to be robust and are detailed in ASTM E2888 (low pH) and ASTM E3042-16 (Triton X-100), accordingly. One of the recent challenges is the need for a replacement of Triton X-100 with a more environmentally friendly detergent with similar log10 reduction value (LRV) achieved. The presentations in this session focused on a detailed assessment of a range of detergents. The most well characterized and potentially robust detergents identified were TDAO (n-Tetradecyl-N,N-dimethylamine-N-oxide) and Simulsol SL 11 W. Key performance factors assessed (in direct comparison with the industry standard Triton X-100) were viral inactivation kinetics (total elapsed time to achieve equilibrium), LRV achieved of enveloped viruses, toxicity, potential impact on product quality and process performance, clearance of residual detergent in subsequent downstream steps, assays to support assessment with appropriate limit of quantification, and commercial supply of detergent of the appropriate quality standard. Both TDAO and Simulsol SL11 had similar overall LRV as Triton-100. In addition, for the low pH viral inactivation, reduced LRV was observed at pH > 3.70 and low salt concentration (outside of the ASTM range), which is a cautionary note when applying low pH inactivation to labile proteins.


Assuntos
Produtos Biológicos , Detergentes , Octoxinol , Inativação de Vírus , Bioensaio
13.
Int J Mol Sci ; 25(7)2024 Mar 25.
Artigo em Inglês | MEDLINE | ID: mdl-38612454

RESUMO

Synucleinopathies are a group of central nervous system pathologies that are characterized by the intracellular accumulation of misfolded and aggregated α-synuclein in proteinaceous depositions known as Lewy Bodies (LBs). The transition of α-synuclein from its physiological to pathological form has been associated with several post-translational modifications such as phosphorylation and an increasing degree of insolubility, which also correlate with disease progression in post-mortem specimens from human patients. Neuronal expression of α-synuclein in model organisms, including Drosophila melanogaster, has been a typical approach employed to study its physiological effects. Biochemical analysis of α-synuclein solubility via high-speed ultracentrifugation with buffers of increasing detergent strength offers a potent method for identification of α-synuclein biochemical properties and the associated pathology stage. Unfortunately, the development of a robust and reproducible method for the evaluation of human α-synuclein solubility isolated from Drosophila tissues has remained elusive. Here, we tested different detergents for their ability to solubilize human α-synuclein carrying the pathological mutation A53T from the brains of aged flies. We also assessed the effect of sonication on the solubility of human α-synuclein and optimized a protocol to discriminate the relative amounts of soluble/insoluble human α-synuclein from dopaminergic neurons of the Drosophila brain. Our data established that, using a 5% SDS buffer, the three-step protocol separates cytosolic soluble, detergent-soluble and insoluble proteins in three sequential fractions according to their chemical properties. This protocol shows that sonication breaks down α-synuclein insoluble complexes from the fly brain, making them soluble in the SDS buffer and thus enriching the detergent-soluble fraction of the protocol.


Assuntos
Sinucleinopatias , Idoso , Animais , Humanos , alfa-Sinucleína , Detergentes , Drosophila melanogaster
14.
BMC Plant Biol ; 24(1): 223, 2024 Mar 27.
Artigo em Inglês | MEDLINE | ID: mdl-38539072

RESUMO

BACKGROUND: Triticale is making its way on dairy farms as an alternative forage crop. This requires the availability of high-yielding triticale varieties with good digestibility. Triticale forage breeding mainly focussed on biomass yield, but efforts to improve digestibility are increasing. We previously investigated the interrelationships among different quality traits in soft dough triticale: starch, acid detergent fibre and in vitro digestibility of organic matter (IVOMD) and of neutral detergent fibre (IVNDFD) of the total plant, IVNDFD and Klason lignin of the stems, and ear proportion and stem length. Here we determine the genetic control of these traits, using a genome-wide association (GWAS) approach. A total of 33,231 DArTseq SNP markers assessed in a collection of 118 winter triticale genotypes, including 101 varieties and 17 breeding lines, were used. RESULTS: The GWAS identified a total of 53 significant marker-trait associations (MTAs). The highest number of significantly associated SNP markers (n = 10) was identified for total plant IVNDFD. A SNP marker on chromosome 1A (4211801_19_C/T; 474,437,796 bp) was found to be significantly associated with ear proportion, and plant and stem IVNDFD, with the largest phenotypic variation for ear proportion (R²p = 0.23). Based on MTAs, candidate genes were identified which were of particular relevance for variation in in vitro digestibility (IVD) because they are putatively involved in plasma membrane transport, cytoskeleton organisation, carbohydrate metabolic processes, protein phosphorylation, and sterol and cell wall biogenesis. Interestingly, a xyloglucan-related candidate gene on chromosome 2R, SECCE2Rv1G0126340, was located in close proximity of a SNP significantly associated with stem IVNDFD. Furthermore, quantitative trait loci previously reported in wheat co-localized with significantly associated SNP markers in triticale. CONCLUSIONS: A collection of 118 winter triticale genotypes combined with DArTseq SNP markers served as a source for identifying 53 MTAs and several candidate genes for forage IVD and related traits through a GWAS approach. Taken together, the results of this study demonstrate that the genetic diversity available in this collection can be further exploited for research and breeding purposes to improve the IVD of triticale forage.


Assuntos
Estudo de Associação Genômica Ampla , Triticale , Detergentes , Melhoramento Vegetal , Fenótipo
15.
Biomolecules ; 14(3)2024 Mar 11.
Artigo em Inglês | MEDLINE | ID: mdl-38540752

RESUMO

Capitellacin is the ß-hairpin membrane-active cationic antimicrobial peptide from the marine polychaeta Capitella teleta. Capitellacin exhibits antibacterial activity, including against drug-resistant strains. To gain insight into the mechanism of capitellacin action, we investigated the structure of the peptide in the membrane-mimicking environment of dodecylphosphocholine (DPC) micelles using high-resolution NMR spectroscopy. In DPC solution, two structural forms of capitellacin were observed: a monomeric ß-hairpin was in equilibrium with a dimer formed by the antiparallel association of the N-terminal ß-strands and stabilized by intermonomer hydrogen bonds and Van der Waals interactions. The thermodynamics of the enthalpy-driven dimerization process was studied by varying the temperature and molar ratios of the peptide to detergent. Cooling the peptide/detergent system promoted capitellacin dimerization. Paramagnetic relaxation enhancement induced by lipid-soluble 12-doxylstearate showed that monomeric and dimeric capitellacin interacted with the surface of the micelle and did not penetrate into the micelle interior, which is consistent with the "carpet" mode of membrane activity. An analysis of the known structures of ß-hairpin AMP dimers showed that their dimerization in a membrane-like environment occurs through the association of polar or weakly hydrophobic surfaces. A comparative analysis of the physicochemical properties of ß-hairpin AMPs revealed that dimer stability and hemolytic activity are positively correlated with surface hydrophobicity. An additional positive correlation was observed between hemolytic activity and AMP charge. The data obtained allowed for the provision of a more accurate description of the mechanism of the oligomerization of ß-structural peptides in biological membranes.


Assuntos
Peptídeos Catiônicos Antimicrobianos , Poliquetos , Animais , Peptídeos Catiônicos Antimicrobianos/farmacologia , Peptídeos Catiônicos Antimicrobianos/química , Dimerização , Micelas , Detergentes , Espectroscopia de Ressonância Magnética , Termodinâmica
16.
J Environ Manage ; 357: 120731, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38552513

RESUMO

Understanding how management influences forage nutritive value and grazer selection within grazing seasons is an ongoing effort for researchers and land managers globally. We used six, 65 ha pastures managed with patch-burn grazing and stocked with either cow-calf pairs (0.45-0.5 ha • AUM-1) or gestating ewes (0.4-0.48 ha • AUM-1) to explore how patterns in rangeland forage drive grazer selection in semi-arid rangeland over four summer grazing seasons at monthly intervals. We used near-infrared spectroscopy to determine nutritive value parameters from monthly forage clippings. We evaluated livestock performance as the average daily weight gains of each animal. We used mixed-effect models and ordination to compare patch and grazer types across the time-since-fire gradient and found that time-since-fire was significant for all measured variables. Cattle and sheep consistently preferred recently burned patches throughout grazing seasons. These recently burned patches typically contained available forage with higher crude protein and moisture content, lower biomass, and lower acid detergent fiber, acid detergent lignin, and neutral detergent fiber compared to intermediate time since fire patches and patches burned three years ago. Differences between patch-burn grazing with cattle and sheep were observed as additional patch contrasts for available biomass and crude protein, but grazer type and ecological site were not statistically significant factors for the nutritive value ordination. Our study indicates that patch-burn grazing is capable of imposing and maintaining heterogeneous, grazer selection, forage biomass, and nutritive value patterns desirable for heterogeneity focused land management, regardless of grazer type. These findings are especially relevant to the northern Great Plains where introduced grasses are homogenizing the structural environment of remaining rangelands. With prescribed fire currently an uncommon practice throughout the region, these findings provide a baseline of expectations for practitioners and land managers implementing patch-burn grazing and illustrate how grazing livestock can benefit from the patch contrast in forage nutritive value and biomass.


Assuntos
Detergentes , Poaceae , Animais , Bovinos , Ovinos , Feminino , Biomassa , Gado , Valor Nutritivo , Estações do Ano , Ração Animal
17.
Curr Microbiol ; 81(5): 114, 2024 Mar 13.
Artigo em Inglês | MEDLINE | ID: mdl-38478167

RESUMO

The moist and warm environment in the household washing machine provides ideal living conditions for the growth and survival of various microorganisms. However, the biodiversity of bacterial community in the different parts of washing machine from Chinese households has not been clarified. In this study, we assessed the bacterial communities in sealing strip, detergent drawer, inner drum, water filter and greywater of ten domestic washing machines quantitatively and qualitatively in Chengdu, southwestern China. The microbial cultivation results indicated that the washing machines from Chengdu had a severe microbial contamination reflected by large counts on bacteria, fungi and coliform. Furthermore, the sequencing data showed that the different parts displayed distinctive bacterial compositions. At the level of genus, the anaerobic bacteria of Caproiciproducens and Acidipropionibacterium were predominant in sealing strip. Barnesiella, Shinella and Sellimonas were detected as the characteristic bacteria in detergent drawer. The pathogens of Luteibacter and Corynebacterium at the genus level were the dominant bacteria in inner drum and water filter, respectively. The genera of Azospira, Roseococcus, Elstera and Aquicella, which belonged to the pathogenic phylum of Proteobacteria, were identified as bioindicators for the greywater. Gene function analysis on the sequencing data illustrated that the bacteria from washing machines were potentially associated with bacterial infectious diseases and antimicrobial resistance. This study shows the bacterial diversity in the different parts of washing machines, providing new clues for bacterial contamination in washing machines from Chinese households.


Assuntos
Bactérias , Detergentes , Proteobactérias , Bacteroidetes , Água
18.
Chemosphere ; 354: 141698, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38490608

RESUMO

The increased use of personal care products and detergents in modern society has raised concerns about their potential adverse effects on the environment. These products contain various chemical compounds that can persist in water bodies, leading to water pollution and ecological disturbances. Bioremediation has emerged as a promising approach to address these challenges, utilizing the natural capabilities of microorganisms to degrade or remove these contaminants. This review examines the current strategies employed in the bioremediation of personal care products and detergents, with a specific focus on their sustainability and environmental impact. This bioremediation is essential for environmental rejuvenation, as it uses living organisms to detergents and other daily used products. Its distinctiveness stems from sustainable, nature-centric ways that provide eco-friendly solutions for pollution eradication and nurturing a healthy planet, all while avoiding copying. Explores the use of microbial consortia, enzyme-based treatments, and novel biotechnological approaches in the context of environmental remediation. Additionally, the ecological implications and long-term sustainability of these strategies are assessed. Understanding the strengths and limitations of these bioremediation techniques is essential for developing effective and environmentally friendly solutions to mitigate the impact of personal care products and detergents on ecosystems.


Assuntos
Cosméticos , Detergentes , Animais , Biodegradação Ambiental , Ecossistema , Estágios do Ciclo de Vida
19.
Plant Physiol Biochem ; 209: 108551, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38537382

RESUMO

Water stress affects the metabolic regulation and delays the growth and development of alfalfa, causing a reduction in biomass. New alfalfa germplasm was created with improved drought tolerance in greenhouse conditions by introducing the key gene P5CDH1 from C. songorica, a xerophytic grass. However, the field adaptability and response mechanism of new drought-tolerant alfalfa germplasms under water stress are still unclear. In the present study, the yield and quality traits of transgenic CsP5CDH1 alfalfa lines under water stress and normal irrigation conditions were measured and analyzed for two years. The genetic variance components of the tested traits were calculated from the data fitted by the mixed linear model. The plant height of all lines showed significant genotypic variation (σ2g) (P < 0.05), and the stem diameter, stem number, and dry weight of all lines had a significant genotype × environment interaction (σ2ge) (P < 0.05). The heritability (H) of plant height, stem diameter, stem number, dry weight and leaf-to-stem ratio of alfalfa lines were 0.87, 0.52, 0.59, 0.52 and 0.50, respectively. There were significant genotype × environment interactions (σ2ge) (P < 0.05) for the quality traits of all lines. The heritabilities (H) of acid detergent fiber and neutral detergent fiber were 0.65 and 0.64, respectively. The results of transcriptional expression analysis with RNA-seq showed that the genes MsProDH1, MsProDH4, MsProDH5, MsP5CDH1, MsP5CS5, MsP5CS9, and MsP5CR1, which are involved in the proline metabolism pathway, played an important role in the drought tolerance of innovative alfalfa germplasm. Under water stress, with the regulation of key genes in the proline metabolism pathway, the proline content of all alfalfa lines increased to varying degrees. Among them, the proline content in the shoots and roots of transgenic line L6 was 7.29 times and 12.22 times that under normal irrigation conditions, respectively. The present study helped to clarify that the new germplasm of alfalfa transformed with the CsP5CDH gene synthesized a large amount of proline under water stress, and effectively slowed leaf water loss, thus improving the drought resistance of alfalfa.


Assuntos
Desidratação , Secas , Medicago sativa/metabolismo , Detergentes/metabolismo , Poaceae , Prolina/metabolismo
20.
Anim Sci J ; 95(1): e13937, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38500367

RESUMO

This experiment aimed to study the degradation characteristics of bagasse after three fermentation treatments in beef cattle. Bagasse 1 was treated with 0.3% lactic acid bacteria (w/w). Bagasse 2 was treated with 0.3% mixed strains (Saccharomyces cerevisiae, Aspergillus niger, Aspergillus oryzae, and lactic acid bacteria at 2:1:1:1). Bagasse 3 was treated with 0.1% cellulase and 0.1% xylanase in addition to 0.3% mixed strains of bagasse 2. The dry matter (DM), crude ash (ASH), crude protein (CP), neutral detergent fiber (NDF), and acid detergent fiber (ADF) in the bagasses were determined. Compared to the control bagasse (without the strain and enzyme treatments), three fermented bagasses showed higher DM after 4 h fermentation. The CP and ASH contents in fermented bagasse 3 were the highest, while the contents of NDF and ADF in fermented bagasse 3 were the lowest among all the groups. The effective degradability of DM, CP, NDF, and ADF was highest in fermented bagasse 3 among the evaluated bagasse feed, followed by fermented bagasse 2 > fermented bagasse 1 > bagasse. Overall, fermented bagasse 3 was better than the control and other treated bagasses, thus fermented bagasse 3 is a hopeful source for ruminant diet of beef cattle.


Assuntos
Celulose , Detergentes , Rúmen , Bovinos , Animais , Rúmen/metabolismo , Fermentação , Detergentes/metabolismo , Digestão , Dieta/veterinária , Ruminantes/metabolismo , Saccharomyces cerevisiae , Ração Animal/análise , Fibras na Dieta/metabolismo
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