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1.
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
2.
Sci Rep ; 14(1): 5715, 2024 03 08.
Artigo em Inglês | MEDLINE | ID: mdl-38459094

RESUMO

Kidney transplantation is a common yet highly demanding medical procedure worldwide, enhancing the quality of life for patients with chronic kidney disease. Despite its prevalence, the procedure faces a shortage of available organs, partly due to contamination by microorganisms, leading to significant organ disposal. This study proposes utilizing photonic techniques associated with organ support machines to prevent patient contamination during kidney transplantation. We implemented a decontamination system using ultraviolet-C (UV-C) irradiation on the preservation solution circulating through pigs' kidneys between harvest and implant. UV-C irradiation, alone or combined with ultrasound (US) and Ps80 detergent during ex-vivo swine organ perfusion in a Lifeport® Kidney Transporter machine, aimed to reduce microbiological load in both fluid and organ. Results show rapid fluid decontamination compared to microorganism release from the organ, with notable retention. By including Ps80 detergent at 0.5% during UV-C irradiation 3 log10 (CFU mL-1) of Staphylococcus aureus bacteria previously retained in the organ were successfully removed, indicating the technique's feasibility and effectiveness.


Assuntos
Descontaminação , Detergentes , Humanos , Animais , Suínos , Descontaminação/métodos , Qualidade de Vida , Diálise Renal , Rim , Preservação de Órgãos/métodos , Perfusão
3.
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
4.
ACS Chem Biol ; 19(3): 763-773, 2024 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-38449446

RESUMO

The inwardly rectifying potassium Kir4.2 channel plays a crucial role in regulating membrane potentials and maintaining potassium homeostasis. Kir4.2 has been implicated in various physiological processes, including insulin secretion, gastric acid regulation, and the pathogenesis of central nervous system diseases. Despite its significance, the number of identified ligands for Kir4.2 remains limited. In this study, we established a method to directly observe ligands avoiding a requirement to observe the high-mass ligand-membrane protein-detergent complexes. This method used collision-induced affinity selection mass spectrometry (CIAS-MS) to identify ligands dissociated from the Kir4.2 channel-detergent complex. The CIAS-MS approach integrated all stages of affinity selection within the mass spectrometer, offering advantages in terms of time efficiency and cost-effectiveness. Additionally, we explored the effect of collisional voltage ramps on the dissociation behavior of the ligand and the ligand at different concentrations, demonstrating dose dependency.


Assuntos
Detergentes , Canais de Potássio Corretores do Fluxo de Internalização , Ligantes , Potássio/metabolismo
5.
J Appl Microbiol ; 135(3)2024 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-38439676

RESUMO

AIMS: We aimed to develop a method to assess the virucidal performance of domestic laundry in a lab-scale washing machine (Rotawash) based on EN 17658. METHODS AND RESULTS: For method development, virus recovery was investigated after drying on cotton carriers for three test viruses murine norovirus (MNV), modified vaccinia virus Ankara (MVA), and bovine coronavirus (BCoV), followed by washing simulations in flasks and Rotawash. MNV and MVA demonstrated sufficient recovery from carriers after drying and washing (up to 40°C and 60 min). BCoV exhibited lower recovery, indicating less relevance as a test virus. Rotawash efficacy tests conducted with MNV, a resistant, non-enveloped virus, showed limited efficacy of a bleach-free detergent, aligning with results from a domestic washing machine. Rotawash washes achieved higher reductions in infectious virus titers than suspension tests, indicating the role of washing mechanics in virus removal. CONCLUSIONS: This study established a practical method to test the virucidal efficacy of laundry detergents in Rotawash, simulating domestic washing.


Assuntos
Detergentes , Norovirus , Bovinos , Animais , Camundongos , Detergentes/farmacologia , Têxteis
6.
J Anim Sci ; 1022024 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-38366060

RESUMO

The purpose of this study was to evaluate the differences in annual pasture and native pasture on dry matter (DM) intake, nutrient digestibility, nitrogen (N) and energy utilization, and methane (CH4) emission of grazing sheep, and to provide the basis for rational livestock grazing in salinized regions. The study used 10 male Hu sheep ♀ × thin-tailed Han sheep ♂ rams (20 ±â€…5 kg) aged 5 mo. Sheep grazing was conducted in annual pasture and native pasture using a 2 × 2 Latin square design. After a 15-d adaptation period for grazing, the digestion and metabolism experiment of sheep were conducted, while CH4 emissions were measured using sulfur hexafluoride tracer gas. DM intake did not differ between annual pasture and native pasture (P = 0.386). Meanwhile, the digestibility of DM (P < 0.001), neutral detergent fiber (P < 0.001), acid detergent fiber (P < 0.01), crude protein (P < 0.001), and ether extract (P < 0.001) of sheep grazing on native pasture was significantly higher than that of annual pasture. Sheep grazing on native pasture had increased N intake (P < 0.001) and N retained (P < 0.001) compared with those grazing on annual pasture. Digestion energy (P < 0.05) and metabolic energy (P < 0.01) of sheep grazing on annual pasture were significantly improved compared with those on native pasture, while fecal energy (P < 0.001), urine energy (P < 0.001) and CH4 energy (CH4-E) output (P < 0.001) and CH4 emission (P < 0.001) of sheep grazing on annual pasture were significantly decreased. The CH4-E/gross energy (GE) values of sheep grazing on annual pasture and native pasture were 0.09 and 0.10, respectively. In conclusion, grazing sheep have higher N utilization on native pasture, whereas grazing sheep have higher energy utilization and low CH4 emissions in annual pasture. In conclusion, annual pasture has a lower CH4-E/GE compared to native pasture, which helps in reducing environmental pollution.


The reduction of methane (CH4) emissions and nitrogen (N) excretion from livestock production systems can help mitigate environmental impact and improve feeding efficiency. The energy requirements of livestock are crucial for enhancing their performance and minimizing environmental impact. It is imperative to accurately ascertain the N and energy efficiency, and CH4 emissions associated with sheep grazing across diverse grassland ecosystems to optimize forage resource utilization without compromising livestock production performance, thereby facilitating sustainable grassland management and grazing practices. Sheep grazing on native pasture had higher nutrient digestibility and N utilization, while sheep grazing on annual pasture showed higher energy utilization and less CH4 emissions. CH4-energy/gross energy for grazing sheep on annual pasture and native pasture was 0.09 and 0.10, respectively. This study assessed the differences in N and energy utilization and CH4 emissions, among sheep grazing on different grasses, providing data support for the development of more rational livestock grazing methods.


Assuntos
Ração Animal , Dieta , Ovinos , Masculino , Animais , Ração Animal/análise , Dieta/veterinária , Metano/metabolismo , Nitrogênio/metabolismo , Detergentes , Carneiro Doméstico , Digestão
7.
Biomolecules ; 14(2)2024 Jan 28.
Artigo em Inglês | MEDLINE | ID: mdl-38397393

RESUMO

The purpose of this review is to succinctly examine the methodologies used in lipid raft research in the brain and to highlight the drawbacks of some investigative approaches. Lipid rafts are biochemically and biophysically different from the bulk membrane. A specific lipid environment within membrane domains provides a harbor for distinct raftophilic proteins, all of which in concert create a specialized platform orchestrating various cellular processes. Studying lipid rafts has proved to be arduous due to their elusive nature, mobility, and constant dynamic reorganization to meet the cellular needs. Studying neuronal lipid rafts is particularly cumbersome due to the immensely complex regional molecular architecture of the central nervous system. Biochemical fractionation, performed with or without detergents, is still the most widely used method to isolate lipid rafts. However, the differences in solubilization when various detergents are used has exposed a dire need to find more reliable methods to study particular rafts. Biochemical methods need to be complemented with other approaches such as live-cell microscopy, imaging mass spectrometry, and the development of specific non-invasive fluorescent probes to obtain a more complete image of raft dynamics and to study the spatio-temporal expression of rafts in live cells.


Assuntos
Detergentes , Microdomínios da Membrana , Microdomínios da Membrana/química , Encéfalo
8.
An Acad Bras Cienc ; 96(1): e20200004, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38359286

RESUMO

This study aimed to evaluate the impact of Meloidogyne javanica, Meloidogyne incognita, and Meloidogyne arenaria on different aspects of the development of sugarcane plants under greenhouse conditions. For this purpose, seedlings of the RB867515 genotype were individually inoculated with 5,000 eggs + second-stage juveniles of their respective nematodes/plant, and non-inoculated plants were used as control. After 330 days of inoculation, the plants were removed from the pots, and the following characteristics were evaluated: fresh mass of the aerial part and root system; leaf area; leaf chlorophyll index; culm diameter; fresh mass of culms; broth volume; contents of neutral and acid detergent fiber, cellulose, hemicellulose, lignin, apparent sucrose in broth, and reducing sugars in broth; total soluble solids concentration. Subsequently, the final population of nematodes in the root system of inoculated plants was determined to calculate the reproduction factor of nematodes. The results showed that all tested Meloidogyne species negatively affected plant development and the composition of some analyzed fractions, in comparison to the non-inoculated control. However, the presence of the root-knot nematode in sugarcane plants increased the contents of neutral and acid detergent fiber, cellulose, hemicellulose, lignin, and reducing sugars, regardless of the Meloidogyne species.


Assuntos
Saccharum , Tylenchoidea , Animais , Lignina , Detergentes , Celulose , Sacarose
9.
J Biotechnol ; 384: 45-54, 2024 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-38403131

RESUMO

Recently developed multi-specific antibody formats enable new therapeutic concepts. Conveniently, formats with an Fc domain allow purification in well-established mAb platform processes. However, due to the structural complexity of the formats, the assembled molecules may be sensitive to extreme pH commonly used for viral inactivation. An alternative to low pH incubation for virus inactivation is the use of a mixture of tri-n-butyl phosphate (TnBP, solvent) and Polysorbate 80 (PS80, detergent). While TnBP is toxic, this combination has a long history of use in the manufacturing of human plasma-derived products that are sensitive to low or high pH incubation. Data are provided demonstrating that the solvent/detergent (S/D) treatment using TnBP and PS80 can be successfully used for pH-sensitive, multi-specific antibody formats in the clarified cell culture fluid (CCCF). A different placement of the S/D within the purification process, namely during the capture by Protein A (PA), has been evaluated. This alternative placement allows effective viral inactivation by S/D while preserving the viral reduction and viral inactivation achieved through the PA step itself, enabling the cumulation of these effects. Furthermore, the process alternative simplifies the liquid handling by reducing the added volumes of the required S/D liquids, thus reducing the amount of toxic TnBP to a minimum. Data are shown demonstrating a complete removal of TnBP and PS80 in the process.


Assuntos
Detergentes , Inativação de Vírus , Humanos , Fator VIII/metabolismo , Anticorpos , Solventes , Concentração de Íons de Hidrogênio
10.
J Anim Sci ; 1022024 Jan 03.
Artigo em Inglês | MEDLINE | ID: mdl-38422238

RESUMO

Two experiments were conducted using 120 pigs to test the hypothesis that supplementation of ß-mannanase could reduce digesta viscosity, enhance nutrient digestion, and improve intestinal health and growth of nursery pigs. In experiment 1, 48 crossbred barrows were randomly allotted to four treatments with increasing levels of ß-mannanase at 0, 200, 400, and 600 U/kg in feeds. All pigs were euthanized on day 12 to collect jejunal digesta to measure digesta viscosity and ileal digesta to measure apparent ileal digestibility (AID) of dry matter (DM), gross energy (GE), neutral detergent fiber (NDF), and acid detergent fiber (ADF). In experiment 2, 72 nursery pigs were randomly allotted to three treatments with increasing levels of ß-mannanase at 0, 400, and 600 U/kg in feeds. Plasma collected on day 9 was used to measure tumor necrosis factor-α (TNF-α), immunoglobulin G (IgG), malondialdehyde (MDA), and protein carbonyl (PC). All pigs were euthanized on day 10 to collect duodenal and jejunal tissues to evaluate the production of TNF-α, IL-6, and MDA, morphology, crypt cell proliferation, and expression of tight junction proteins in the jejunum. Data were analyzed using the MIXED procedure for polynomial contrasts and the NLMIXED procedure for broken-line analysis of SAS. In experiment 1, ß-mannanase supplementation tended to have quadratic effects on digesta viscosity (P = 0.085) and AID of GE (P = 0.093) in the pigs. In experiment 2, jejunal digesta viscosity of the pigs was reduced (P < 0.05) when ß-mannanase was supplemented at 360 U/kg of feed. ß-Mannanase supplementation linearly reduced (P < 0.05) TNF-α, IgG, MDA, and PC in the duodenum, and TNF-α, IgG, and MDA in the jejunum of the pigs. ß-Mannanase supplementation linearly increased (P < 0.05) villus height to crypt depth ratio and crypt cell proliferation in the jejunum. ß-Mannanase supplementation tended to linearly improve (P = 0.083) expression of zonula occludens-1 in the jejunum. In conclusion, supplementation of ß-mannanase at 360 U/kg reduced the digesta viscosity and up to 600 U/kg positively affected intestinal health and growth of pigs by reducing inflammation and oxidative stress whilst enhancing structure and barrier function in the jejunum.


Nursery pigs face challenges in digesting complex carbohydrates in their feeds, which can negatively affect their growth and intestinal health. In particular, ß-mannans can increase digesta viscosity and hinder nutrient digestion of nursery pigs. ß-Mannanase, an enzyme that breaks down ß-mannans, has been used in nursery feeds to alleviate negative impacts on nutrient utilization and intestinal health of nursery pigs. This study investigated the effects of increasing supplementation levels of ß-mannanase on intestinal health, nutrient utilization, and growth of nursery pigs. The results showed that supplementation of ß-mannanase at 360 U/kg in the feed reduced the digesta viscosity in the jejunum and up to 600 U/kg positively had beneficial effects on the intestinal health and growth of nursery pigs by reducing inflammation and oxidative stress through improving structure and barrier function in the jejunum.


Assuntos
Dieta , beta-Manosidase , Animais , Suínos , Dieta/veterinária , beta-Manosidase/farmacologia , Fator de Necrose Tumoral alfa , Detergentes/farmacologia , Digestão , Suplementos Nutricionais/análise , Imunoglobulina G , Ração Animal/análise
11.
Animal ; 18(3): 101084, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38367312

RESUMO

Procedures such as transport and marketing can subject animals to water and feed deprivation and impair animal health and performance. Maintaining the mineral status of animals under these conditions can bring benefits to health and performance. The use of hydroxychloride mineral sources can improve mineral status, nutrient digestibility and performance. Two studies were conducted to investigate how the supplementation of 02 trace mineral sources of Cu and Zn and 48-hour water/feed deprivation would affect the performance and metabolism of grass-fed beef cattle. In the first study, 20 castrated and rumen-canulated Nellore steers (BW = 350 ±â€¯132 kg; 20 m) were distributed in individual pens, in a 2 × 2 factorial arrangement: supplemental Cu and Zn sources from inorganic vs hydroxychloride (HTM) and 48-hours deprivation (WFD) vs unrestricted (WFU) access to water and feed. The 57d of study was divided into two periods: (1) Adaptation from -21d to -1d and (2) evaluation from 0d to 36d. Interaction between deprivation × period was detected (P < 0.05) for digestibility of DM (DMD), organic matter (OMD), neutral detergent fiber (NDFD), and acid detergent fiber (ADFD). Deprivation increased DMD, OMD, NDFD, and ADFD immediately after the deprivation period (3-5d), but impaired digestibility at longer periods such as 11-13d and 32-34d. DM (DMI) and nutrient intake (P = 0.075), as well as NDFD were higher in HTM. Several ruminal parameters were affected by deprivation: short-chain fatty acids concentration decreased, while rumen pH increased (deprivation × time; P < 0.05); decreased propionate, butyrate and increased isobutyrate, isovalerate, and valerate in WFD (deprivation × time; P < 0.05), respectively. In the second study, eighty-four intact Nellore males (BW = 260 ±â€¯35 kg) were blocked by BW and randomly assigned to Urochloa brizantha cv. Marandu paddocks for 131d in a 2 × 2 factorial arrangement. Liver Cu was higher in WFU/HTM animals (mineral × deprivation; P < 0.05). Interaction between deprivation × period (P < 0.05) was detected for BW and average daily gain (ADG). On 2d and 12d after deprivation, WFD increased ADG and recovered the BW lost. In conclusion, water and feed deprivation imposed in these trials were able to impact several nutrient digestibility and ruminal fermentation parameters in short- and long-term. Performance was not affected by the studied factors. Furthermore, supplementation with sources of Cu and Zn hydroxychloride increased Cu in the liver and tended to increase DMI and NDFD.


Assuntos
Cobre , Zinco , Masculino , Bovinos , Animais , Suplementos Nutricionais , Dieta/veterinária , Água/metabolismo , Detergentes/metabolismo , Digestão , Minerais/metabolismo , Fermentação , Ração Animal/análise , Rúmen/metabolismo
12.
Biofabrication ; 16(2)2024 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-38394679

RESUMO

Decellularized matrices are an attractive choice of scaffold in regenerative medicine as they can provide the necessary extracellular matrix (ECM) components, signals and mechanical properties. Various detergent-based protocols have already been proposed for decellularization of skeletal muscle tissue. However, a proper comparison is difficult due to differences in species, muscle origin and sample sizes. Moreover, a thorough evaluation of the remaining acellular matrix is often lacking. We compared an in-house developed decellularization protocol to four previously published methods in a standardized manner. Porcine skeletal muscle samples with uniform thickness were subjected to in-depth histological, ultrastructural, biochemical and biomechanical analysis. In addition, 2D and three-dimensional cytocompatibility experiments were performed. We found that the decellularization methods had a differential effect on the properties of the resulting acellular matrices. Sodium deoxycholate combined with deoxyribonuclease I was not an effective method for decellularizing thick skeletal muscle tissue. Triton X-100 in combination with trypsin, on the other hand, removed nuclear material but not cytoplasmic proteins at low concentrations. Moreover, it led to significant alterations in the biomechanical properties. Finally, sodium dodecyl sulphate (SDS) seemed most promising, resulting in a drastic decrease in DNA content without major effects on the ECM composition and biomechanical properties. Moreover, cell attachment and metabolic activity were also found to be the highest on samples decellularized with SDS. Through a newly proposed standardized analysis, we provide a comprehensive understanding of the impact of different decellularizing agents on the structure and composition of skeletal muscle. Evaluation of nuclear content as well as ECM composition, biomechanical properties and cell growth are important parameters to assess. SDS comes forward as a detergent with the best balance between all measured parameters and holds the most promise for decellularization of skeletal muscle tissue.


Assuntos
Detergentes , Matriz Extracelular , Animais , Suínos , Detergentes/química , Detergentes/metabolismo , Detergentes/farmacologia , Matriz Extracelular/metabolismo , Octoxinol/química , Octoxinol/metabolismo , Octoxinol/farmacologia , Músculo Esquelético , Dodecilsulfato de Sódio/química , Dodecilsulfato de Sódio/metabolismo , Dodecilsulfato de Sódio/farmacologia , Tecidos Suporte , Engenharia Tecidual/métodos
13.
Bioconjug Chem ; 35(2): 223-231, 2024 02 21.
Artigo em Inglês | MEDLINE | ID: mdl-38215010

RESUMO

Membrane protein structures are essential for the molecular understanding of diverse cellular processes and drug discovery. Detergents are not only widely used to extract membrane proteins from membranes but also utilized to preserve native protein structures in aqueous solution. However, micelles formed by conventional detergents are suboptimal for membrane protein stabilization, necessitating the development of novel amphiphilic molecules with enhanced protein stabilization efficacy. In this study, we prepared two sets of tandem malonate-derived glucoside (TMG) variants, both of which were designed to increase the alkyl chain density in micelle interiors. The alkyl chain density was modulated either by reducing the spacer length (TMG-Ms) or by introducing an additional alkyl chain between the two alkyl chains of the original TMGs (TMG-Ps). When evaluated with a few membrane proteins including a G protein-coupled receptor, TMG-P10,8 was found to be substantially more efficient at extracting membrane proteins and also effective at preserving protein integrity in the long term compared to the previously described TMG-A13. This result reveals that inserting an additional alkyl chain between the two existing alkyl chains is an effective way to optimize detergent properties for membrane protein study. This new biochemical tool and the design principle described have the potential to facilitate membrane protein structure determination.


Assuntos
Detergentes , Proteínas de Membrana , Proteínas de Membrana/metabolismo , Detergentes/química , Micelas
14.
Sci Rep ; 14(1): 1660, 2024 01 18.
Artigo em Inglês | MEDLINE | ID: mdl-38238544

RESUMO

The patch-clamp technique has revolutionized neurophysiology by allowing to study single neuronal excitability, synaptic connectivity, morphology, and the transcriptomic profile. However, the throughput in recordings is limited because of the manual replacement of patch-pipettes after each attempt which are often also unsuccessful. This has been overcome by automated cleaning the tips in detergent solutions, allowing to reuse the pipette for further recordings. Here, we developed a novel method of automated cleaning by sonicating the tips within the bath solution wherein the cells are placed, reducing the risk of contaminating the bath solution or internal solution of the recording pipette by any detergent and avoiding the necessity of a separate chamber for cleaning. We showed that the patch-pipettes can be used consecutively at least ten times and that the cleaning process does not negatively impact neither the brain slices nor other patched neurons. This method, combined with automated patch-clamp, highly improves the throughput for single and especially multiple recordings.


Assuntos
Detergentes , Ultrassom , Neurônios/fisiologia , Neurofisiologia , Técnicas de Patch-Clamp
15.
J Anim Sci ; 1022024 Jan 03.
Artigo em Inglês | MEDLINE | ID: mdl-38290533

RESUMO

An experiment was conducted to determine the effects of betaine on growth performance and intestinal health in rabbits fed diets with different levels of digestible energy. During a 36-d experiment, a total of 144 healthy 35-d-old weaned New Zealand white rabbits with a similar initial body weight (771.05 ±â€…41.79 g) were randomly distributed to a 2 × 3 factorial arrangement. Experimental treatments consisted of two levels of digestible energy (normal: 10.20 and low: 9.60 MJ/kg) and three levels of betaine (0, 500, and 1,000 mg/kg). Results indicated that rabbits fed the diet with low digestible energy (LDE) had reduced body gain/feed intake on days 1 to 14 and 1 to 36 (P < 0.05), increased the apparent total tract digestibility (ATTD) of neutral detergent fiber, acid detergent fiber (ADF), and n-free extract, and decreased the ATTD of gross energy (GE), crude fiber, and organic matter (OM; P < 0.05). The LDE diet upregulated the gene abundance levels of duodenum junctional adhesion molecule-3 (JAM-3) and downregulated the ileum toll-like receptor 4, myeloid differentiation factor 88, and interleukin-6 (IL-6; P < 0.05). Activities of amylase, lipase, trypsin, and the immunoglobulin M content in the jejunum were decreased in the LDE treatment group (P < 0.05). Dietary betaine supplementation increased the ATTD of GE, dry matter (DM), ADF, and n-free extract by LDE (P < 0.05). The villus height, crypt depth, and goblet cell numbers were decreased, and the villus-crypt ratio was increased in the duodenum (P < 0.05). The gene abundance levels of duodenum IL-2 were downregulated, and the duodenum JAM-2 and JAM-3 were upregulated (P < 0.05). Furthermore, the addition of betaine to the LDE diet increased the ATTD of GE, DM, and OM in rabbits (P < 0.05). Gene abundance levels of ileum IL-6 and duodenum JAM-3 were upregulated (P < 0.05). In summary, LDE diets can reduce the activity of intestinal digestive enzymes and decrease the ATTD of nutrients. However, the addition of betaine to LDE diets improved the intestinal barrier structure and nutrient ATTD in rabbits, with better results when betaine was added at an additive level of 500 mg/kg.


Insufficient dietary energy can cause many negative effects on animal production and cause intestinal diseases, which are one of the main causes of morbidity and mortality in rabbits. Results of some experiments demonstrated that betaine has various physiological functions such as improving energy utilization and intestinal health. Therefore, the aim of this study was to evaluate the effects of betaine supplementation on growth performance, intestinal function, and health in rabbits fed diets with different levels of digestible energy. The results showed that the addition of betaine to a low-digestible energy diet improved the gut barrier structure and nutrient digestibility in rabbits.


Assuntos
Betaína , Detergentes , Coelhos , Animais , Betaína/farmacologia , Detergentes/farmacologia , Interleucina-6 , Digestão , Dieta/veterinária
16.
Int J Biol Macromol ; 260(Pt 1): 129507, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38244731

RESUMO

Halophiles are excellent sources of detergent proteases that are attributed to stability in alkaline pH, salts, surfactants, and hydrophobic solvents. The lower enzymatic yields and tedious downstream processes necessitate the search for newer halophilic sources. We have previously reported a halotolerant Exiguobacterium sp. TBG-PICH-001, which secretes solvent-tolerant alkaline protease/s. The present study describes the heterologous expression of two protease genes, namely, rsep metalloprotease (WP_195864791, 1.23 Kb) and tpa serine protease (WP_195864453, 0.879 Kb) genes. These were cloned into the pET 22b + plasmid vector and expressed in Escherichia coli BL21(DE3). The recombinant proteases rsep and tpa showed respective yields of 6.3 and 6.7 IU/mg, 11 and 12-fold higher than the crude native protease/s from TBG-PICH-001. These showed soluble expression at 46 and 32 KDa, respectively. These were purified to homogeneity through Ni-NTA-affinity chromatography. The purified proteases were characterized for properties like pH & temperature optima and stability, substrate specificity, kinetic parameters, and detergent attributes. They showed affinity towards various substrates with a respective Km of 392 and 301 µM towards casein. The recombinant proteases exhibited stability in the alkaline pH (7-10), surfactants, metal ions, detergents, and hydrophobic solvents, rendering their suitability as detergent additives.


Assuntos
Detergentes , Exiguobacterium , Exiguobacterium/metabolismo , Detergentes/química , Solventes/química , Estabilidade Enzimática , Serina Proteases/química , Tensoativos , Temperatura , Concentração de Íons de Hidrogênio , Proteínas de Bactérias/química
17.
Anal Chem ; 96(6): 2574-2581, 2024 02 13.
Artigo em Inglês | MEDLINE | ID: mdl-38291764

RESUMO

Mass spectrometry (MS) analysis is often challenged by contaminations from detergents, salts, and polymers that compromise data quality and can damage the chromatography and MS instruments. However, researchers often discover contamination issues only after they acquire the data. There is no existing contaminant assay that is sensitive enough to detect trace amounts of contaminants from a few microliters of samples prior to MS analysis. To address this crucial need in the field, we developed a sensitive, rapid, and cost-effective contaminant spot check and removal assay (ContamSPOT) to detect and quantify trace amounts of contaminants, such as detergents, salts, and other chemicals commonly used in the MS sample preparation workflow. Only 1 µL of the sample was used prior to MS injection to quantify contaminants by ContamSPOT colorimetric or fluorometric assay on a thin layer chromatography (TLC) plate. We also optimized contaminant removal methods to salvage samples with minimal loss when ContamSPOT showed a positive result. ContamSPOT was then successfully applied to evaluate commonly used bottom-up proteomic methods regarding the effectiveness of removing detergent, peptide recovery, reproducibility, and proteome coverage. We expect ContamSPOT to be widely adopted by MS laboratories as a last-step quality checkpoint prior to MS injection. We provided a practical decision tree and a step-by-step protocol with a troubleshooting guide to facilitate the use of ContamSPOT by other researchers. ContamSPOT can also provide a unique readout of sample cleanliness for developing new MS-based sample preparation methods in the future.


Assuntos
Detergentes , Proteômica , Detergentes/química , Proteômica/métodos , Reprodutibilidade dos Testes , Sais , Espectrometria de Massas/métodos
18.
Chemphyschem ; 25(4): e202300824, 2024 Feb 16.
Artigo em Inglês | MEDLINE | ID: mdl-38179823

RESUMO

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.


Assuntos
Resíduos de Praguicidas , Resíduos de Praguicidas/análise , Detergentes/análise , Frutas/química , Verduras/química , Contaminação de Alimentos/análise
19.
Environ Sci Technol ; 58(3): 1741-1751, 2024 Jan 23.
Artigo em Inglês | MEDLINE | ID: mdl-38184844

RESUMO

An anaerobic dynamic membrane bioreactor (AnDMBR) mimicking rumen conditions was developed to enhance the hydrolysis of lignocellulosic materials and the production of volatile fatty acids (VFAs) when treating food waste. The AnDMBR was inoculated with cow rumen content and operated at a 0.5 day hydraulic retention time, 2-4 day solids retention time, a temperature of 39 °C, and a pH of 6.3, characteristics similar to those of a rumen. Removal rates of neutral detergent fiber and acid detergent fiber of 58.9 ± 8.4 and 69.0 ± 8.6%, respectively, and a VFA yield of 0.55 ± 0.12 g VFA as chemical oxygen demand g volatile solids (VS)fed-1 were observed at an organic loading rate of 18 ± 2 kg VS m-3 day-1. The composition and activity of the microbial community remained consistent after biofilm disruption, bioreactor upset, and reinoculation. Up to 66.7 ± 5.7% of the active microbial populations and 51.0 ± 7.0% of the total microbial populations present in the rumen-mimicking AnDMBR originated from the inoculum. This study offers a strategy to leverage the features of a rumen; the AnDMBR achieved high hydrolysis and fermentation rates even when treating substrates different from those fed to ruminants.


Assuntos
Alimentos , Eliminação de Resíduos , Bovinos , Animais , Feminino , Anaerobiose , Biomassa , Rúmen , Hidrólise , Detergentes , Reatores Biológicos , Fermentação , Ácidos Graxos Voláteis
20.
Vet Med Sci ; 10(1): e31347, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-38227709

RESUMO

BACKGROUND: The use of plants and by-products, which are containing a high amount of secondary and anti-nutritional compounds such as tannins, in animal feed is limited. The methods that can reduce these compounds make facilitate their use in animal feed. OBJECTIVES: The aim of this study was to reduce the adverse effects of pomegranate peel (PP) tannin for fattening lambs using the tannase-producing bacteria. METHODS: Twenty-one Arabi male lambs (averagely 35 ± 3.8 kg weight and 8 ± 1.0 months age) were used in a completely randomized design with three treatments and seven replications in the present experiment. The experimental treatments included 1 - control diet (CNT, no PP), 2 - diet containing untreated PP (raw PP, UTPP) and 3 - diet containing PP treated with tannase-producing bacteria (bacteria treating PP, BTPP). RESULTS: Using UTPP decreased nutrient intake compared to the control and treatment with tannase-producing bacteria again significantly increased nutrient intake compared to the UTPP (p < 0.05). The digestibilities of organic matter, neutral detergent fibre and acid detergent fibre in the control treatment were significantly higher than UTPP and BTPP and in the BTPP were significantly higher than the UTPP (p < 0.05). The use of UTPP in the diet significantly decreased the pH, ammonia nitrogen concentration and the total protozoa population of the rumen compared to the control (p < 0.05), and treatment with bacteria increased them again. The lowest total protozoa population was observed in UTPP treatments (p < 0.05). The highest concentration of blood glucose was observed in UTPP; however, the highest concentrations of blood urea nitrogen, cholesterol, triglyceride, high-density lipoprotein (non-significant) and low-density lipoprotein were in the control treatment. The effect of experimental treatments on the dry matter consumption of the whole period was significant; however, there was no significant effect on average daily gain, feed conversion ratio, feed efficiency and longissimus muscle colorimetric systems. CONCLUSIONS: Therefore, considering the positive effects of treatment PP with tannin-degrading bacteria relative to raw PP, using these bacteria is a proper way to reduce tannin, thus improving the nutritional value of PP for ruminants.


Assuntos
Compostos de Boro , Hidrolases de Éster Carboxílico , Punica granatum , Porfirinas , Taninos , Animais , Detergentes , Digestão , Valor Nutritivo , Rúmen/fisiologia , Ovinos , Carneiro Doméstico , Taninos/análise , Taninos/metabolismo , Masculino
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