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Foods (Basel, Switzerland) May 2024Bee products are considered true wonders of nature, used since ancient times, and studied even today for their various biological activities. In this study, we...
Bee products are considered true wonders of nature, used since ancient times, and studied even today for their various biological activities. In this study, we hypothesise that Romanian bee products from different origins (micro apiary products, lyophilised forms, commercial) exhibit distinct chemical compositions, influencing their biological activities. An LC-MS analysis revealed varied polyphenolic content patterns, with cumaric acid, ferulic acid, rosmarinic acid, and quercitine identified in significant amounts across all samples. Primary anti-inflammatory evaluation phases, including the inhibition of haemolysis values and protein denaturation, unveiled a range of protective effects on red blood cells (RBC) and blood proteins, contingent upon the sample concentration. Antimicrobial activity assessments against 12 ATCC strains and 6 pathogenic isolates demonstrated varying efficacy, with propolis samples showing low efficacy, royal jelly forms displaying moderate effectiveness, and apilarnin forms exhibiting good inhibitory activity, mostly against Gram-positive bacteria. Notably, the lyophilised form emerged as the most promising sample, yielding the best results across the biological activities assessed. Furthermore, molecular docking was employed to elucidate the inhibitory potential of compounds identified from these bee products by targeting putative bacterial and fungal proteins. Results from the docking analysis showed rosmarinic and rutin exhibited strong binding energies and interactions with the putative antimicrobial proteins of bacteria (-9.7 kcal/mol to -7.6 kcal/mol) and fungi (-9.5 kcal/mol to -8.1 kcal/mol). The findings in this study support the use of bee products for antimicrobial purposes in a biologically active and eco-friendly proportion while providing valuable insights into their mechanism of action.
PubMed: 38790755
DOI: 10.3390/foods13101455 -
Antioxidants (Basel, Switzerland) May 2024Alcohol is toxic to neurons and can trigger alcohol-related brain damage, neuronal loss, and cognitive decline. Neuronal cells may be vulnerable to alcohol toxicity and...
Alcohol is toxic to neurons and can trigger alcohol-related brain damage, neuronal loss, and cognitive decline. Neuronal cells may be vulnerable to alcohol toxicity and damage from oxidative stress after differentiation. To consider this further, the toxicity of alcohol to undifferentiated SH-SY5Y cells was compared with that of cells that had been acutely differentiated. Cells were exposed to alcohol over a concentration range of 0-200 mM for up to 24 h and alcohol effects on cell viability were evaluated via MTT and LDH assays. Effects on mitochondrial morphology were examined via transmission electron microscopy, and mitochondrial functionality was examined using measurements of ATP and the production of reactive oxygen species (ROS). Alcohol reduced cell viability and depleted ATP levels in a concentration- and exposure duration-dependent manner, with undifferentiated cells more vulnerable to toxicity. Alcohol exposure resulted in neurite retraction, altered mitochondrial morphology, and increased the levels of ROS in proportion to alcohol concentration; these peaked after 3 and 6 h exposures and were significantly higher in differentiated cells. Protein carbonyl content (PCC) lagged behind ROS production and peaked after 12 and 24 h, increasing in proportion to alcohol concentration, with higher levels in differentiated cells. Carbonylated proteins were characterised by their denatured molecular weights and overlapped with those from adult post-mortem brain tissue, with levels of PCC higher in alcoholic subjects than matched controls. Hence, alcohol can potentially trigger cell and tissue damage from oxidative stress and the accumulation of oxidatively damaged proteins.
PubMed: 38790685
DOI: 10.3390/antiox13050580 -
Scientific Reports May 2024Oral disorders can exert systemic ramifications beyond their localized effects on dental tissues, implicating a wide array of physiological conditions. The utilization...
Oral disorders can exert systemic ramifications beyond their localized effects on dental tissues, implicating a wide array of physiological conditions. The utilization of essential oils (EOs) for protection of oral health represents a longstanding practice. Consequently, in this investigation, essential oil derived from Nigella sativa seeds (NSEO) underwent isolation via the hydro-distillation process, followed by a comprehensive evaluation of its antioxidant, anti-inflammatory, anti-fungal, antibacterial activities, and cytocompatibility. The isolated NSEO manifested as a pale-yellow substance and was found to harbor a diverse spectrum of bioactive constituents, including steroids, triterpenoids, flavonoids, phenols, proteins, alkaloids, tannin, sesquiterpenoid hydrocarbons, monoterpenoid alcohol, and monoterpenoid ketone (thymoquinone). Notably, the total phenolic content (TPC) and total flavonoid content (TFC) of NSEO were quantified at 641.23 μg GAE/gm and 442.25 μg QE/g, respectively. Furthermore, NSEO exhibited concentration-dependent inhibition of protein denaturation, HRBC membrane stabilization, and hemolysis inhibition. Comparative analysis revealed that NSEO and chlorhexidine (CHX) 0.2% displayed substantial inhibition of hemolysis compared to aspirin. While NSEO and CHX 0.2% demonstrated analogous antibacterial activity against Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa, NSEO showcased heightened efficacy against Lactobacillus acidophilus and Candida albicans. Additionally, NSEO exhibited pronounced effects against periodontal pathogens such as Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, and Prevotella intermedia. Importantly, no cytotoxicity was observed on human gingival fibroblast cell lines. These findings underscore the potential of NSEO as a potent antibacterial and antifungal agent in the management of oral microbial pathogens, thereby offering avenues for the development of innovative therapies targeting diverse oral inflammatory conditions. Nevertheless, further investigations are imperative to unlock its full therapeutic repertoire.
Topics: Oils, Volatile; Antioxidants; Nigella sativa; Anti-Inflammatory Agents; Humans; Microbial Sensitivity Tests; Anti-Bacterial Agents; Seeds; Anti-Infective Agents
PubMed: 38789533
DOI: 10.1038/s41598-024-62915-1 -
Journal of Dairy Science May 2024Residual lipids (RL) in whey protein isolate (WPI) are detrimental to optimal functional applications (like foaming and low turbidity) and contribute to off-flavor...
Residual lipids (RL) in whey protein isolate (WPI) are detrimental to optimal functional applications (like foaming and low turbidity) and contribute to off-flavor development during powder storage. The objective of this research was to prepare an experimental WPI by removing RL without using the traditional microfiltration (MF) process and compare its properties with commercially available WPIs made using MF and some other whey powders. We hypothesize that by adjusting the pH of whey to < 5.0, we would be close to the isoelectric point of any remaining denatured proteins (DP) and phospholipoproteins (PLP), and therefore reduce electrostatic repulsion between these molecules. Further, demineralization of the acidified whey protein solution by ultrafiltration (UF) combined with diafiltration (DF) should reduce ionic hindrance to aggregation and thereby help with the aggregation of these denatured proteins as well as most RL; centrifugation or clarification could be used to remove these materials. Calcium should also be more extensively removed by this approach, which should improve the heat stability of the experimental WPI. Demineralization was achieved on a pilot scale by acidifying liquid (cheese) whey protein concentrate (WPC-34) to pH 4.5 using HCl, and UF the whey protein solution along with extensive DF using acidified (pH∼3.5) reverse osmosis filtered (RO) water. Demineralized whey protein solution was adjusted to various combinations of pH (4.1 to 4.9), conductivities (500 to 2000 μS.cm), and protein concentrations (1 to 7%) and then centrifuged at 10,000 × g for 10 min. The effective sedimentation (precipitation) of RL in these treatments was estimated by measuring the turbidity of the supernatants. Maximum precipitation was observed at pH 4.5-4.7. Reducing conductivity via UF/DF increased the precipitation of RL due to reduced ionic hindrance to aggregation Maximum sedimentation of RL was observed at protein concentrations ≤3% because of a higher density difference between the precipitate and serum phase. SDS-PAGE analysis confirmed the sedimentation of PLPs, caseins, and DPs upon isoelectric precipitation at pH ∼4.5, while native whey proteins or undenatured whey proteins remained soluble in the supernatant, unaffected by the pretreatment. To scale up the process, 750 L of fluid WPC34 was acidified and demineralized by UF (volume concentration factor = 1.35) and DF until the permeate solids reached 0.1% (when desired demineralization was achieved), clarified using a pilot-scale desludging clarifier to remove RL, neutralized, ultrafiltered to concentrate the protein, and then spray-dried to produce an experimental WPI (91% protein and 1.8% fat db). In another trial, demineralized UF concentrate was clarified by gravity sedimentation and the supernatant was neutralized, ultrafiltered, and spray-dried to produce a second experimental WPI (91% protein and < 1% fat db). These experimental WPI powders were compared with several commercially available WPI powders to assess functional properties like solubility, heat stability, foamability and foam strength, gelation, and sensory attributes over accelerated storage. Experimental WPI had excellent functional properties, had low turbidity, were highly heat stable and only developed very slight to slight off-flavors upon accelerated storage, their properties were comparable to the WPI manufactured commercially using MF even after accelerated storage.
PubMed: 38788845
DOI: 10.3168/jds.2024-24671 -
Journal of the American Chemical Society Jun 2024High-resolution structural NMR analyses of membrane proteins are challenging due to their large size, resulting in broad resonances and strong signal overlap. Among the...
High-resolution structural NMR analyses of membrane proteins are challenging due to their large size, resulting in broad resonances and strong signal overlap. Among the isotope labeling methods that can remedy this situation, segmental isotope labeling is a suitable strategy to simplify NMR spectra and retain high-resolution structural information. However, protein ligation within integral membrane proteins is complicated since the hydrophobic protein fragments are insoluble, and the removal of ligation side-products is elaborate. Here, we show that a stabilized split-intein system can be used for rapid and high-yield protein trans-splicing of integral membrane proteins under denaturing conditions. This setup enables segmental isotope labeling experiments within folded protein domains for NMR studies. We show that high-quality NMR spectra of markedly reduced complexity can be obtained in detergent micelles and lipid nanodiscs. Of note, the nanodisc insertion step specifically selects for the ligated and correctly folded membrane protein and simultaneously removes ligation byproducts. Using this tailored workflow, we show that high-resolution NMR structure determination is strongly facilitated with just two segmentally isotope-labeled membrane protein samples. The presented method will be broadly applicable to structural and dynamical investigations of (membrane-) proteins and their complexes by solution and solid-state NMR but also other structural methods where segmental labeling is beneficial.
Topics: Membrane Proteins; Isotope Labeling; Nuclear Magnetic Resonance, Biomolecular
PubMed: 38787792
DOI: 10.1021/jacs.4c03294 -
Metabolites May 2024The tomato is a fruit vegetable rich in nutritional and medicinal value grown in greenhouses and fields worldwide. It is severely sensitive to heat stress, which... (Review)
Review
The tomato is a fruit vegetable rich in nutritional and medicinal value grown in greenhouses and fields worldwide. It is severely sensitive to heat stress, which frequently occurs with rising global warming. Predictions indicate a 0.2 °C increase in average surface temperatures per decade for the next three decades, which underlines the threat of austere heat stress in the future. Previous studies have reported that heat stress adversely affects tomato growth, limits nutrient availability, hammers photosynthesis, disrupts reproduction, denatures proteins, upsets signaling pathways, and damages cell membranes. The overproduction of reactive oxygen species in response to heat stress is toxic to tomato plants. The negative consequences of heat stress on the tomato have been the focus of much investigation, resulting in the emergence of several therapeutic interventions. However, a considerable distance remains to be covered to develop tomato varieties that are tolerant to current heat stress and durable in the perspective of increasing global warming. This current review provides a critical analysis of the heat stress consequences on the tomato in the context of global warming, its innate response to heat stress, and the elucidation of domains characterized by a scarcity of knowledge, along with potential avenues for enhancing sustainable tolerance against heat stress through the involvement of diverse advanced technologies. The particular mechanism underlying thermotolerance remains indeterminate and requires further elucidatory investigation. The precise roles and interplay of signaling pathways in response to heat stress remain unresolved. The etiology of tomato plants' physiological and molecular responses against heat stress remains unexplained. Utilizing modern functional genomics techniques, including transcriptomics, proteomics, and metabolomics, can assist in identifying potential candidate proteins, metabolites, genes, gene networks, and signaling pathways contributing to tomato stress tolerance. Improving tomato tolerance against heat stress urges a comprehensive and combined strategy including modern techniques, the latest apparatuses, speedy breeding, physiology, and molecular markers to regulate their physiological, molecular, and biochemical reactions.
PubMed: 38786760
DOI: 10.3390/metabo14050283 -
Gels (Basel, Switzerland) May 2024To determine the evolution of microbial community and microbial shift under anaerobic processes, this study investigates the use of denaturing gradient gel...
To determine the evolution of microbial community and microbial shift under anaerobic processes, this study investigates the use of denaturing gradient gel electrophoresis (DGGE). In the DGGE, short- and medium-sized DNA fragments are separated based on their melting characteristics, and this technique is used in this study to understand the dominant bacterial community in mesophilic and thermophilic anaerobic digestion processes. Dairy manure is known for emitting greenhouse gases (GHGs) such as methane, and GHG emissions from manure is a biological process that is largely dependent on the manure conditions, microbial community presence in manure, and their functions. Additional efforts are needed to understand the GHG emissions from manure and develop control strategies to minimize the biological GHG emissions from manure. To study the microbial shift during anaerobic processes responsible for GHG emission, we conducted a series of manure anaerobic digestion experiments, and these experiments were conducted in lab-scale reactors operated under various temperature conditions (28 °C, 36 °C, 44 °C, and 52 °C). We examined the third variable region (V3) of the 16S rRNA gene fingerprints of bacterial presence in anaerobic environment by PCR amplification and DGGE separation. Results showed that bacterial community was affected by the temperature conditions and anaerobic incubation time of manure. The microbial community structure of the original manure changed over time during anaerobic processes, and the community composition changed substantially with the temperature of the anaerobic process. At Day 0, the sequence similarity confirmed that most of the bacteria were similar (>95%) to sp. (strain: ATCC 31012), a Gram-negative bacteria, regardless of temperature conditions. At day 7, the sequence similarity of DNA fragments of reactors (28 °C) was similar to sp.; however, the DNA fragments of effluent of reactors at 44 °C and 52 °C were similar to (strain: DSM 5265) (similarity: 97%) and (strain: DSM 16624) (similarity: 100%), respectively. At day 60, the analysis showed that DNA fragments of effluent of 28 °C reactor were similar to (strain: NBRC 10162) (similarity: 87%), and DNA fragments of effluent of 36 °C reactors were similar to (strain: GB8-1) (similarity: 91%). In reactors with a relatively higher temperature, the DNA fragments of effluent of 44 °C reactor were similar to (strain: JC13) (similarity: 86%), and the DNA fragments of effluent of 52 °C reactor were similar to (strain: DSM 5265) (similarity: 99%). To authors' knowledge, this is one of the few studies where DGGE-based approach is utilized to study and compare microbial shifts under mesophilic and thermophilic anaerobic digestions of manure simultaneously. While there were challenges in identifying the bands during gradient gel electrophoresis, the joint use of DGGE and sequencing tool can be potentially useful for illustrating and comparing the change in microbial community structure under complex anaerobic processes and functionality of microbes for understanding the consequential GHG emissions from manure.
PubMed: 38786256
DOI: 10.3390/gels10050339 -
Gels (Basel, Switzerland) May 2024This study aimed to investigate the gelling behavior of faba bean (FB) and chickpea (CP) flour between 10 and 20% () concentration at pH 3.0, 5.0, and 7.0. Both sources...
This study aimed to investigate the gelling behavior of faba bean (FB) and chickpea (CP) flour between 10 and 20% () concentration at pH 3.0, 5.0, and 7.0. Both sources formed at pH 3.0 and 5.0 self-standing gels with 12% () of flour, while 16% () of flour was required to obtain a gel at pH 7.0. During gelling between 40 and 70 °C, a sharp increase of the elastic modulus G' was observed in both flours, mainly due to water absorption and swelling of the starch, one of the major constituents in the ingredients. Increasing the temperature at 95 °C, G' increased due to the denaturation of globulins and therefore the exposure of their internal part, which allowed more hydrophobic interactions and the formation of the gel. After cooling, both FB and CP gels displayed a solid-like behavior (tan δ ranging between 0.11 and 0.18) with G' values at pH 3.0 and 5.0 significantly ( < 0.05) higher than those at pH 7.0, due to the lower electrostatic repulsions at pHs far from the isoelectric point. The rheological properties were supported by the water binding capacity values, confirming the better gels' strength described by rheological analysis. These results will enhance our understanding of the role of legume flours in formulating innovative and sustainable food products as alternatives to animal ones.
PubMed: 38786226
DOI: 10.3390/gels10050309 -
Frontiers in Oncology 2024For liquid biopsy of cancer, the extraction of circulating cell-free DNA (cfDNA) from plasma is required. We evaluated the efficacy of use of magnetic submicron...
OBJECTIVE
For liquid biopsy of cancer, the extraction of circulating cell-free DNA (cfDNA) from plasma is required. We evaluated the efficacy of use of magnetic submicron particles coated with abundant small zwitterions (MSP-ZEWBs) for extracting short fragments of cfDNA.
METHODS
We developed and optimized an MSP-ZEWB-based cfDNA extraction method using ampholytic ion-exchange materials and compared its results with those using a control kit. We measured the cfDNA concentration by quantitative polymerase-chain-reaction and using the Qubit method and analyzed cfDNA fragmentation patterns using a bioanalyzer.
RESULTS
The fragment size of cfDNA isolated from glycine hydrochloric acid at a pH of 2.2 exhibited a better alignment with the DNA marker. The highest DNA intensity was observed at the final concentration of 0.8% polyethylene glycol 8000. The intensity of cfDNA decreased significantly when isolated from plasma with DNA marker using MSP-ZEWBs with an adsorption buffer containing guanidine hydrochloride or isothiocyanoguanidine. All fragments were successfully extracted using MSP-ZEWBs from both plasma and phosphate-buffered saline. Notably, the intensity of short cfDNA fragments isolated using MSP-ZEWBs remained consistent for recovery of long DNA fragments. indicating a potential selective of small fragments.
CONCLUSION
The extraction of plasma cfDNA with MSP-ZEWBs requires no protein denaturation, shows resistance to cells remaining in plasma, and demonstrates higher overall efficiency and better reproducibility than other extraction methods. Use of MSP-ZEWBs may greatly enhance liquid biopsy of cancers through the analysis of plasma cfDNA in clinical practice.
PubMed: 38779084
DOI: 10.3389/fonc.2024.1397680 -
Saudi Journal of Biological Sciences Jul 2024Protein glycation, hyper-inflammatory reactions, and oxidative stress play a crucial role in the pathophysiology of numerous diseases. The current work evaluated the...
Protein glycation, hyper-inflammatory reactions, and oxidative stress play a crucial role in the pathophysiology of numerous diseases. The current work evaluated the protective ability of ethyl alcohol extract of leaves from holy basil ( Linn) against inflammation, oxidative stress, glycation and advanced glycation endproducts formation. Various assays assessed prementioned properties of holy basil. In addition, molecular docking was conducted. The highest hydrogen peroxide reduction activity (72.7 %) and maximum percentage of DPPH scavenging (71.3 %) depicted its vigorous antioxidant abilities. Furthermore, it showed the most excellent protection against proteinase activity (67.247 %), prevention of denaturation of egg albumin (65.29 %), and BSA (bovine serum albumin) (68.87 %) with 600 µg/ml. Percent aggregation index (57.528 %), browning intensity (56.61 %), and amyloid structure (57.0 %) were all reduced significantly using 600 μg/ml of extract. Additionally, the antimicrobial potential was also confirmed. According to a molecular docking study, active leaf extract ingredients were found to bind with superoxide dismutase, catalase, and carbonic anhydrase. As a conclusion, has a variety of health-promoting properties that may reduce the severity of many diseases in diabetic patients. However, in order to ascertain the mechanisms of action of the components of its leaves in disease prevention, more thorough research based on pharmacological aspects is needed.
PubMed: 38766504
DOI: 10.1016/j.sjbs.2024.104003