Metabolic Activity Research Articles

In vitro study of essential oils encapsulated in chitosan microparticles against Histoplasma capsulatum and their pathogenicity in Caenorhabditis elegans.

Histoplasmosis, caused by Histoplasma capsulatum, poses risks for immunocompromised individuals. With limited therapeutic options, this study explores microparticles as antimicrobial delivery systems for Cymbopogon flexuosus and Pelargonium graveolens essential oils against H. capsulatum. The broth microdilution assay showed MICs of 32 to 128 µg/mL in filamentous phase and 8 to 64 µg/mL in yeast phase. Combining microparticles with antifungal drugs demonstrated synergistic effects in both filamentous and yeast-like forms with amphotericin B or itraconazole. Chitosan microparticles reduced H. capsulatum biofilm biomass and metabolic activity by about 60% at 512 µg/mL. In vivo evaluation with Caenorhabditis elegans showed H. capsulatum caused over 90% mortality. These findings highlight the potential use of chitosan microparticles as a delivery system for essential oils against H. capsulatum, especially in combination with other compounds.

Bumble bee gut microbial community structure differs between species and commercial suppliers, but metabolic potential remains largely consistent.

Bumble bees are key pollinators for natural and agricultural plant communities. Their health and performance are supported by a core gut microbiota composed of a few bacterial taxa. However, the taxonomic composition and community structure of bumble bee gut microbiotas can vary with bee species, environment, and origin (i.e., whether colonies come from the wild or a commercial rearing facility), and it is unclear whether metabolic capabilities therefore vary as well. Here we used metagenomic sequencing to examine gut microbiota community composition, structure, and metabolic potential across bumble bees from two different commercial Bombus impatiens suppliers, wild B. impatiens, and three other wild bumble bee species sampled from sites within the native range of all four species. We found that the community structure of gut microbiotas varied between bumble bee species, between populations from different origins within species, and between commercial suppliers. Notably, we found that Apibacter is consistently present in some wild bumble bee species-suggesting it may be a previously unrecognized core phylotype of bumble bees-and that commercial B. impatiens colonies can lack core phylotypes consistently found in wild populations. However, despite variation in community structure, the high-level metabolic potential of gut microbiotas was largely consistent across all hosts, including for metabolic capabilities related to host performance, though metabolic activity remains to be investigated.IMPORTANCEOur study is the first to compare genome-level taxonomic structure and metabolic potential of whole bumble bee gut microbiotas between commercial suppliers and between commercial and wild populations. In addition, we profiled the full gut microbiotas of three wild bumble bee species for the first time. Overall, our results provide new insight into bumble bee gut microbiota community structure and function and will help researchers evaluate how well studies conducted in one bumble bee population will translate to other populations and species. Research on taxonomic and metabolic variation in bumble bee gut microbiotas across species and origins is of increasing relevance as we continue to discover new ways that social bee gut microbiotas influence host health, and as some bumble bee species decline in range and abundance.

Regulatory mechanism of carbohydrate metabolism pathways on oil biosynthesis of oil plant Symplocos paniculata

The mechanism underlying oil synthesis in oil plant fruits remains elusive, as sugar metabolism provides the essential carbon skeleton without a clear understanding of its intricate workings. The transcriptome and oil and sugar metabolites’ content of Symplocos paniculate, an extraordinary oil plant with immense ecological significance, were subjected to a comparative analysis throughout fruit development. The findings unveiled that the impact of sugar metabolism on oil synthesis varied throughout distinct stages of fruit development. Remarkably, during the initial phase of fruit development from 10 to 90 days after flowering (DAF), pivotal genes involved in starch biosynthesis, such as ADP-glucose pyrophosphorylase (AGP), starch synthase (SS), and starch branching enzyme (SBE), facilitated an earlier accumulation of starch within the fruit. Whereas, during the fruit maturation stage (from 90 DAF to 170 DAF), the expression of phosphofructokinase 1 (PFK-1), pyruvate kinase (PK) and pyruvate dehydrogenase (PDH) enzyme genes involved in the glycolysis pathway was significantly upregulated, thereby facilitating a rapid and substantial accumulation of oil. The sugar metabolism activity of S. paniculata fruit exerts a crucial influence on the process of oil synthesis, which is highly dependent on the specific developmental stage. These significant discoveries provide potential candidate genes for advanced genetic improvement using molecular biotechnology, thus enhancing both fruit oil production and modifying the composition of fatty acids.

The Effect of Environmental Factors on the Nitrate and Nitrite Metabolism of Oral Actinomyces and Schaalia Species.

Actinomyces naeslundii and Schaalia odontolytica belong to the most predominant nitrite-producing bacteria in the oral microbiome. Nitrite has antibacterial and vasodilatory effects that may contribute to maintaining oral and systemic health. We have previously elucidated the metabolic characteristics of the nitrite-producing activity of oral Veillonella species and the effects of oral environmental factors. However, this is still unknown for Actinomyces and Schaalia species. Furthermore, these bacteria are thought to degrade nitrite. Therefore, this study aimed to comprehensively elucidate the effects of environmental factors (pH, oxygen concentration, glucose, lactate, and the presence of nitrate/nitrite during growth) on nitrate and nitrite metabolism of these bacterial species using the type strains. Nitrite was quantified by Griess reagent, and final metabolites were analyzed by high-performance liquid chromatography (HPLC). The nitrite-producing activity of A. naeslundii and S. odontolytica was affected variously by environmental factors. Especially in A. naeslundii, under anaerobic conditions, the activity increased in a concentration-dependent manner with the addition of glucose or lactate and was higher at lower pH when lactate was added. The nitrite-degrading activity of both bacteria was lower than the nitrite-producing activity and was less affected by environmental factors. Metabolites from glucose by A. naeslundii were different with and without nitrate, suggesting that nitrate altered metabolic pathways. The growth was inhibited under anaerobic conditions but promoted under aerobic conditions. These results indicate that the nitrite-producing capacity of the oral microflora must take into account not only the composition and abundance of bacteria but also the variation in metabolic activity due to various environmental factors.

Efficacy of nitrofuran derivatives against biofilms of Histoplasma capsulatum strains and their in vivo toxicity.

To evaluate the efficacy of two nitrofuran derivatives against biofilms formed by two strains of Histoplasma capsulatum and to study the toxicity of these compounds in alternative models: Caenorhabditis elegans, Galleria mellonella, and zebrafish. The metabolic activity of biofilms was measured after treatment using the XTT reduction assay. Scanning electron microscopy (SEM) and confocal microscopy were used to observe damage to mature biofilms. Survival curves were generated for G. mellonella, while percentage survival was determined for C. elegans and zebrafish. The compounds showed efficacy against early and mature biofilms at concentrations equal to or up to two times higher than those required to eliminate planktonic fungal cells (3.90 to 31.25 μg/mL). Micrographs showed a reduction in metabolic activity, biofilm thickness, and extracellular matrix. In addition, the compounds showed little or no toxicity in alternative models, even at the highest concentrations tested. These results are promising for the development of new therapeutic alternatives, especially for species, such as H. capsulatum, which are recognized as high-priority pathogens. Few studies have investigated resistance and antifungal treatment targeting biofilms of this species, making this work a relevant contribution to future approaches.

Photobiostimulation of Saccharomyces cerevisiae with Nano Cobalt Ferrite: A Sustainable Approach to Bioethanol Production from Banana Peels.

This study presents a pioneering investigation into the effects of cobalt ferrite nanoparticles (CoFe₂O₄ NPs) on the fermentation efficiency of Saccharomyces cerevisiae and the production of bioethanol from banana peel biomass. The findings reveal a notable difference between the control sample, which produced only 11.16% bioethanol, and the enhanced yield achieved with the addition of 100 ppm CoFe₂O₄ nanoparticles, which reached an impressive 52.16%. This substantial increase underscores the potential of nanomaterials to catalyze fermentation processes, likely due to their unique physicochemical properties that enhance metabolic activity in yeast cells. Additionally, the study explored the impact of visible light irradiation on bioethanol production. Light exposure alone resulted in a 15.44% increase in ethanol yield compared to the control sample without nanoparticles. This emphasizes the role of light in enhancing fermentation dynamics, potentially by providing additional energy for metabolic reactions. When CoFe₂O₄ nanoparticles were activated by visible light, their stimulating effects on ethanol production were further intensified, leading to a remarkable ethanol yield of 63.01%. These results indicate a synergistic relationship between the nanoparticles and light, where the photoactivation of the nanomaterials not only boosts their catalytic properties but also enhances the overall metabolic activity of Saccharomyces cerevisiae. This suggests a promising approach for developing more efficient fermentation processes, potentially increasing bioethanol yields while utilizing waste materials. Overall, the integration of nanotechnology with renewable biomass resources offers a viable pathway toward more sustainable energy solutions.

Uncovering proteome variations and concomitant quality changes of different drying methods Cordyceps sinensis by 4D-DIA structural proteomics

IntroductionCordyceps sinensis is a fungus, serves dual purposes as both a medicinal herb and a food source. Due to its high water content, fresh Cordyceps sinensis is difficult to preserve, necessitating the drying necessary to process Cordyceps sinensis.MethodsUsing 4D-DIA proteomics, researchers analyzed the proteome profiles of fresh Cordyceps sinensis (CK) under three different drying conditions: vacuum freeze-drying (FD), oven-drying (OD), and air-drying (AD). In addition, it was found that the protein and free sulfhydryl content of Cordyceps sinensis decreased significantly and the disulfide bond content increased after different drying methods.Results and discussionA total of 3762 proteins were identified, showing variations between groups and high protein content. In the control groups consisting of fresh Cordyceps sinensis samples and the three drying methods, FD. vs CK exhibited the fewest differentially abundant proteins, with the majority being upregulated. On the other hand, CK vs OD displayed the greatest amount of distinct proteins, with a significant rise in both up-regulated and down-regulated proteins. Analysis of KEGG indicated that the distinct proteins were predominantly concentrated in pathways like the ribosome, synthesis of coenzymes, and metabolism of amino sugar and nucleotide sugar. Notably, there was a significant overlap between ribosome and ribosome biogenesis in eukaryotes pathways. The process of drying Cordyceps sinensis resulted in a significant upregulation of the expression of proteins linked to various metabolic pathways. This observation suggests that the drying treatment might activate or enhance certain biochemical processes within the organism, potentially influencing its overall metabolic activity. This finding highlights the importance of post-harvest dry methods on the biochemical properties of Cordyceps sinensis, which could have implications for its nutritional and medicinal value.This study provides a theoretical basis for the realization of Cordyceps sinensis resource utilization and storage methods, and provides theoretical support for guaranteeing the sustainable development of Cordyceps sinensis resources.

GDH1-dependent α-ketoglutarate promotes HBV transcription by modulating histone methylations on the cccDNA minichromosome.

Hepatitis B virus (HBV) hijacks host cell metabolism, especially host glutamine metabolism, to support its replication. Glutamate dehydrogenase 1 (GDH1), a mitochondrial enzyme crucial for glutamine metabolism, can interact with histone demethylases to regulate gene expression through histone methylation. However, the mechanisms underlying GDH1-mediated glutamine metabolism reprogramming and the roles of key metabolites during HBV infection remain unclear. Transcriptomic and metabolomic analyses of HBV-infected cell were performed. Both HBV-infected cells and humanized liver chimeric mice were used to elucidate the effect of glutamine metabolism on HBV. HBV infection leads to the abnormal activation of glutamine metabolism, including upregulation of key enzymes and metabolites involved in glutamine metabolism. The viral core protein (HBc) mediates the translocation of GDH1 into the nucleus, where GDH1 activates covalently closed circular DNA (cccDNA) transcription by converting glutamate to α-ketoglutarate (αKG). Mechanistically, the promoting effect of GDH1-derived αKG on cccDNA transcription is independent of its conventional role. Rather, αKG directly interacts with the lysine-specific demethylase KDM4A and enhances KDM4A demethylase activity to regulate αKG-dependent histone demethylation, controlling cccDNA transcription. Our findings highlight the importance of glutamine metabolism in HBV transcription and suggest that glutamine deprivation is a potential strategy for silencing cccDNA transcription.

Tailoring the Electrocatalytic Properties of Porphyrin Covalent Organic Frameworks for Highly Selective Oxygen Sensing In Vivo.

In vivo selective sensing of oxygen (O2) dynamics in the central nervous system could provide insights into energy metabolism and neural activities. Although the electrocatalytic four-electron oxygen reduction reaction (ORR) paves an effective way to the electrochemical sensing of O2 in vivo, the concurrent hydrogen peroxide reduction reaction (HPRR) within the potential windows for four-electron ORR unfortunately poses a great challenge to the conventional mechanism employed for selective electrochemical O2 sensing. In this work, we find that regulation of the linkers within the skeleton of porphyrin-based covalent organic frameworks (COFs) could improve the selectivity of the O2 sensor against hydrogen peroxide (H2O2). The electrochemical results reveal that the Co porphyrin active sites facilitate the direct four-electron pathway for ORR and that the Co porphyrin-based COF, enriched with pyrene units, shows enhanced four-electron ORR kinetics and better tolerance to HPRR. The theoretical calculation suggests that introducing pyrene units essentially weakens the adsorption of H2O2, leading to suppression of the HPRR. The microsensor fabricated with the Co porphyrin-based COF as the electrocatalyst features a high selectivity for real-time monitoring of O2 in a living rat brain.

Electrode- and Label-Free Assessment of Electrophysiological Firing Rates through Cytochrome C Monitoring via Raman Spectroscopy.

In vitro neurotoxicology aims to assess and predict the side effects of exogenous chemicals toward the human brain. Among the exploited approaches, electrophysiological techniques stand out for the high spatiotemporal resolution and sensitivity, with the patch clamp considered the gold standard technique for such purposes. However, structural toxicity and metabolic effects may elude detection when only the electrical activity is measured, highlighting the need for integrating electrophysiological recordings with complementary approaches such as optical methods. In this study, we describe an integrated platform for recording neuronal electrical activity and performing chemical analysis with a noninvasive label-free optical imaging, Raman spectroscopy. Specifically, we developed a protocol that maximizes the signal-to-noise ratio while avoiding the crosstalk of the electrical and spectroscopical readouts and any phototoxicity associated with the laser exposure. Synchronous and sequential electrical-optical measurements were carried out and compared, with the sequential approach being more suitable for the longitudinal investigation and correlation of the neuronal electrical activity to the intracellular content of reduced cytochrome C, lipids, proteins, and nucleic acids. Data analysis shows a strong correlation between the metabolic status of the single cells and the overall neuronal firing rate, suggesting the electrode- and label-free assessment of the neuronal firing rates through the monitoring of cytochrome C via Raman spectroscopy when multielectrode array devices with high electrical noise and impedance are used. Conversely, the neuronal firing rate and the reduced cytochrome C content were not correlated to lipids, proteins, and nucleic acids. Thus, this study demonstrates the crosstalk of the neuronal firing rate and reduced cytochrome C as downstream and upstream features of the neuronal metabolic activity and that through the monitoring of the de novo synthesis of lipids, proteins, and nucleic acids, Raman spectroscopy provides additional information for a more accurate assessment of the acute and chronic neurotoxicity.

The effect of culture media and seeding densities on the human malignant testicular germ cell line (NT2/D1) growth pattern

Testicular cancer is a common malignancy in young males, comprising 5% of urological cancers and 1% of all male cancers. The NT2/D1 cell line, a pluripotent embryonal carcinoma, is valuable in cancer and developmental biology research due to its differentiation potential into neurons and glial cells. Although Fetal Bovine Serum (FBS) is often used to promote cell growth, its limitations necessitate optimized culture conditions. Monitoring cell growth and health through methods such as cell counting, viability assays, growth curves, and microscopy is crucial for reliable research, particularly for evaluating specific cell characteristics before in vitro studies. The study compared the growth patterns of NT2/D1 cell lines under different culture conditions and seeding densities using trypan blue cell counting and MTT methods. The NT2/D1 cell lines cultured in heat-inactivated FBS (H.iFBS) and non-heat-inactivated FBS (Non-H.iFBS) exhibit distinct growth patterns, doubling times, and cell morphologies. Cell density and the condition of the culture medium are two key parameters influencing metabolic activities and enhancing variability. There is no preference between culturing cells in different FBS conditions yet it should be considered in experiment design. Thus systematic reporting of all cell culture parameters is essential to facilitate better comparisons of in vitro and in vivo experimental data.

Tetramycin ameliorates tebuconazole·azoxystrobin to control leaf spot and viral diseases of Taizishen

Leaf spot and viral diseases are the most frequently occurring leaf problems in Taizishen production. In this study, we examined the controlling role played by the co-application of tetramycin and low dose tebuconazole·azoxystrobin against leaf spot and viral diseases in Taizishen, investigating its resistance, electrophysiological information, growth and quality. Among them, electrophysiological information indicators include electrical signals [intrinsic capacitance (IC), resistance (IR), impedance (IZ), capacitive reactance (IXc), and inductive reactance (IXL)], intracellular water metabolism, nutrient transport, and plant metabolic activity. The results indicate that 0.3% tetramycin 1000-time + 75% tebuconazole·azoxystrobin 2000-time diluent controlled leaf spot and viral diseases the best, with protection effects of 90.03%~90.46% and 71.67%~73.08% at 15~30 days after the last fungicide application, respectively. These values are obviously higher than those treated with high doses of tetramycin or tebuconazole·azoxystrobin alone. Concurrently, their combined application could notably enhance total soluble flavonoids, total soluble phenols, protective enzyme activity, IC, intracellular water metabolism, nutrient transport, and metabolic activity, while reducing its MDA, IR, IZ, IXc, and IXL. Moreover, their co-application also could obviously ameliorate photosynthesis, biomass, agronomic trait, and root growth and quality, as well as actually reduce tebuconazole·azoxystrobin input. Additionally, the control effects of leaf spot and viral diseases in Taizishen treated by their combined application exhibited significant correlations with its disease resistance, electrophysiology, photosynthesis, growth, and quality parameters. This study highlights the combined application of low-dosage tebuconazole·azoxystrobin and tetramycin as a practicable measure for controlling leaf spot and viral diseases in Taizishen, promoting its resistance, growth, and quality, as well as reducing chemical pesticide application.

Antimicrobial effects of chlorhexidine gel associated with hyaluronic acid or cetylpyridinium chloride in a multispecies biofilm

Chlorhexidine (CHX) and cetylpyridinium chloride (CPC) are commonly used as mouthwashes due to their antimicrobial effect. More recently, hyaluronic acid (HA) has also been associated to oral health products aiming to improve their anti-inflammatory and antimicrobial effects. This study aimed to evaluate the effect of three different CHX-based commercially available products on the subgingival microbial composition and metabolic activity using a multispecies biofilm model. A biofilm model composed of 33 bacterial species with 7 days of maturation on a Calgary plate device was used. The multispecies biofilm was treated with CHX 0.2% (positive control), CHX 0.2% + CPC (test 1), CHX 0.2% + HA 1% (test 2), and culture medium (negative control). The metabolic activity of the multispecies biofilm was measured using a spectrophotometric assay and the microbial composition by checkerboard DNA-DNA hybridization. The studied groups were compared using ANOVA and post hoc Bonferroni tests. The significance level was established at 5%. The CHX+CPC gel was more effective than the other treatments in reducing proportions of red complex species and increasing bacterial species associated with periodontal health (p<0.05). A reduction of approximately 54% was observed in the microbial metabolic activity of biofilms treated with CHX and CHX + CPC, and 26% in biofilms treated with CHX+HA. The CHX+CPC gel seems to have a superior antibacterial potential when compared to a gel containing CHX only, or CHX+HA, in an in vitro multispecies biofilm model.

Single-cell profiling reveals a reduced epithelial defense system, decreased immune responses and the immune regulatory roles of different fibroblast subpopulations in chronic atrophic gastritis

PurposeTo identify key cellular changes and molecular events in atrophic mucosa, we aimed to elucidate the molecular mechanisms driving the occurrence of chronic atrophic gastritis (CAG).MethodsWe used single-cell RNA sequencing (scRNA-seq) to characterize changes in the epithelial state and tissue microenvironment associated with CAG. The molecular changes were identified by comparing differentially expressed genes (DEGs) between the two mucosa states. Gene Ontology (GO) pathway enrichment analysis was used to explore the potential functional changes in each cell subtype in atrophic mucosa. Gene set score analysis was conducted to compare the functional roles of different fibroblast subtypes and functional changes in cell subtypes between the CAG and control groups. Metabolic analysis was performed to compare the metabolic activity of C1Q+ macrophages under different conditions. NichNet analysis was used to analyze the regulatory relationships between CCL11+APOE+ fibroblasts and C1Q+ macrophages and between CCL11+APOE+ fibroblasts and CD8+ effector T cells. Transcription factor (TF) analysis was performed to determine the transcription status of different T-cell subtypes in atrophic and normal mucosa.ResultsWe generated a single-cell transcriptomic atlas from 3 CAG biopsy samples and paired adjacent normal tissues. Our analysis revealed that chief cells and parietal cells exhibited a loss of detoxification ability and that surface mucous cells displayed a reduced antimicrobial defense ability in CAG lesions. The mucous neck cells in CAG lesions showed upregulation of genes related to cell cycle transition, which may lead to aberrant DNA replication. Additionally, cells with the T exhaustion phenotype infiltrated under CAG condition. C1Q+ macrophages exhibited reduced phagocytosis, downregulated expression of pattern recognition receptors and decreased metabolic activity. NichNet analysis revealed that a subpopulation of CXCL11+APOE+ fibroblasts regulated the inflammatory response in the pathogenesis of atrophic gastritis. APSN+CXCL11+APOE+ fibroblasts were found to be associated with gastric cancer (GC) development.ConclusionsThe main goal of this study was to comprehensively elucidate the cellular changes in CAG lesions. We observed an immune decline in the mucosal microenvironment during the development of CAG, including a reduced immune response of C1Q+ macrophages, reduced cytotoxicity of T cells, and increased infiltration of exhausted T cells. Specifically, we demonstrated that different epithelial subtypes aberrantly express genes related to susceptibility to external bacterial infection and aberrant cell cycle progression. Our study provides new insights into the functions of epithelial changes and immune alterations during the development of CAG.

Targeting AI-2 quorum sensing: harnessing natural products against Streptococcus suis biofilm infection

The biofilm acts as a protective layer for Streptococcus suis (S. suis), contributing to the development of drug resistance and chronic infections. Autoinducer 2 (AI-2) quorum sensing represents the primary regulatory pathway governing biofilm formation in S. suis. Consequently, targeting AI-2 quorum sensing to inhibit biofilm formation represents a promising strategy for preventing and managing drug resistance and chronic infections caused by S. suis. This study established a small natural product library by integrating commercial drug molecules with Chinese herbal medicine molecules. Consequently, two natural products, salvianolic acid A (SAA) and rhapontin (RH), which target S. suis AI-2 via quorum sensing, were identified. SAA and RH inhibit AI-2 synthesis through noncompetitive and competitive binding to S-ribosylhomocysteinase (LuxS). By inhibiting S. suis AI-2 quorum sensing, these compounds modulate the expression of adhesion genes and the synthesis of extracellular polysaccharides (EPS), reducing the adhesion ability of S. suis and ultimately inhibiting biofilm formation. Using LC‒MS/MS, we further analysed the impact of SAA and RH on the metabolic activity of S. suis, revealing the potential medicinal value of these compounds. Finally, the efficacy of SAA and RH against S. suis infection was validated in Galleria mellonella larvae, confirming their significant anti-infection effects.

Evaluation of selected indigenous spices- and herbs-derived small molecules as potential inhibitors of SREBP and its implications for breast cancer using MD simulations and MMPBSA calculations.

In this study, we conducted an extensive analysis of 252 bioactive compounds derived from native spices and herbs for their potential anti-breast cancer activity against sterol regulatory element-binding protein (SREBP), using in silico techniques. To evaluate the oral bioavailability, overall pharmacokinetics, and safety profiles of these compounds, we employed Lipinski's rule of five and ADME descriptors, which depicted 66 lead molecules. These molecules were then docked with the SREBP using molecular docking tools, which revealed that diosgenin and smilagenin were the most promising hits compared to the reference inhibitor betulin, with average binding affinities of -7.42 and -7.37kcal/mol and -6.27kcal/mol, respectively. To further assess the stability of these complexes along with betulin, we conducted molecular dynamics simulations over a 100ns simulation period. We employed various parameters, including the root-mean-square deviation, root-mean-square fluctuation, solvent-accessible surface area, free energy of solvation, and radius of gyration, followed by principal component analysis. Furthermore, we evaluated the toxicity of the selected compounds against various anticancer cell lines, as well as their metabolic activity related to CYP450 metabolism and biological activity spectrum. Based on these results, both molecules exhibited promising drug candidate potential and could be utilized for further experimental analysis to elucidate their anticancer potential.

Differential Signaling Pathways Identified in Aqueous Humor, Anterior Capsule, and Crystalline Lens of Age-Related, Diabetic, and Post-Vitrectomy Cataract

Background: The purpose of this study was to detect proteomic alterations and corresponding signaling pathways involved in the formation of age-related cataract (ARC), diabetic cataract (DC), and post-vitrectomy cataract (PVC). Methods: Three sample types, the aqueous humor (AH), the anterior capsule (AC), and the content of the phaco cassette, were collected during phacoemulsification surgery. The samples were obtained from 12 participants without diabetes mellitus (DM), 11 participants with DM, and 7 participants without DM, with a history of vitrectomy surgery in the past 12 months. The Sp3 protocol (Single-Pot, Solid-Phase, Sample-Preparation) was used for the sample preparation. The recognition and quantification of proteins were carried out with liquid chromatography online with tandem mass spectrometry. The DIA-NN software was applied for the identification and quantification of peptides/proteins. Statistical analysis and data visualization were conducted on Perseus software. Data are available via ProteomeXchange. Results: A very rich atlas of the lens and AH proteome has been generated. Glycosaminoglycan biosynthesis and the non-canonical Wnt receptor signaling pathway were differentially expressed in ARC compared to both the DC and PVC groups. In the PVC group, complement activation was differentially expressed in AH samples, while glutathione metabolism and oxidoreductase activity were differentially expressed in AC samples. Microfilament motor activity, microtubule cytoskeleton organization, and microtubule binding were differentially expressed in the DC and PVC groups in both AH and AC samples. Conclusions: The results of this study expand the existing knowledge on pathways involved in the pathophysiology of cataract, and suggest possible important druggable targets for slower progression or even prevention of cataract.

Omega-3 polyunsaturated fatty acids modify glucose metabolism in THP-1 monocytes.

Chronic inflammation is a driving factor in diseases like obesity and type 2 diabetes. Enhanced cellular glucose metabolism may contribute to heightened immune activation. A human supplementation trial showed that the n-3 PUFA α-linolenic acid (ALA) reduced oxidative phosphorylation in monocytes. Our objective here is to assess the direct effects of ALA and docosahexaenoic acid (DHA) on glucose metabolism in a cell culture model and to explore possible mechanisms. THP-1 monocytes were treated with 10-40 μM of ALA or DHA and compared with vehicle and oleic acid controls. The Seahorse XFe24 and Oroboros O2k Oxygraph systems were used to approximate catabolic rates in the presence of glucose. Both ALA and DHA reduced oxidative phosphorylation. We identified pyruvate dehydrogenase kinase 4 (PDK4) as a possible mechanistic candidate explaining the effect of DHA. Additionally, both n-3 PUFAs reduced LPS-induced IL-1β production, while only DHA increased reactive oxygen species to a small but significant extent. Our data suggest that ALA and DHA trigger a re-wiring of bioenergetic pathways in monocytes, possibly via the upregulation of PKD4. Given the close relationship between cell metabolism and immune cell activation, this may represent a novel mechanism by which n-3 PUFAs modulate immune function and inflammation.

Role of metabolism, resistance, and/or antagonism as drivers of endomicrobiomes assemblage in Origanum heracleoticum L

The understanding of selective forces driving the compartmentalization of microbiota in plants remains limited. In this study, we performed a phenotypic characterization of bacterial endophytes isolated from the medicinal plant Origanum heracleoticum, together with the determination of the antibiotic resistance profiles and the antagonistic interactions of communities within and across different plant organs. Results revealed organ-related differences in the metabolic capabilities of bacteria, with those associated with stems displaying the highest metabolic activity for carbon sources. Contrarily, the patterns of antibiotic resistance appeared closely aligned with the taxonomical classification of the endophytes. The presence of antagonistic interactions, likely spurred by resource limitations, favor bacteria exhibiting greater metabolic plasticity. In conclusion, this research advances our comprehension of the intricate dynamics between plants and their associated microbiota, indicating that its composition is mainly influenced by forces contributing to the selection of distinct functions and phenotypic traits.

Comparative Evaluation of the Biocompatibility of Sponges Based on Different Chitosan Salts in vitro

INTRODUCTION. The relevance of the study is due to the increase in the number of human skin lesions as a result of natural and man-made emergencies. Among the natural polymers widely used for the manufacture of wound dressings is chitosan, but its poor solubility is a technological and biological limitation for its effective inclusion and use as a component of wound dressings. One way to solve the problem is to modify chitosan with glycolic, lactic or ascorbic acids. For eff ective and adequate use of chemical modifications of chitosan as a material for wound dressings, it is necessary to evaluate their biocompatibility. AIM. In vitro assessment of the cytotoxicity and hemostatic properties of sponges based on chitosan modified with glycolic, lactic and ascorbic acids. MATERIALS AND METHODS. Sponges based on water-soluble chitosan salts were provided by Chemical Company “Orion” LTD. The biocompatibility of the obtained products was assessed on the culture of mesenchymal stem cells, using light and fluorescent microscopy. The metabolic activity of the cells (MTT-test) and the sorption properties of the biomaterials were assessed by spectrophotometry. RESULTS AND DISCUSSION. Chitosan glycolate or chitosan lactate sponges ensure the survival of most adherent cells, while chitosan ascorbate sponges inhibit cell viability and growth. It was found that chitosan glycolate sponges have the highest blood sorption capacity and are capable of absorbing over 700 μl of blood/cm3. Chitosan lactate and chitosan ascorbate sponges demonstrate a sorption capacity of 250 μl of blood/cm3. However, the ascorbate-based material inhibits the metabolic activity of cells. CONCLUSION. Based on the research results, sponges made of chitosan glycolate or lactate, which have good biocompatibility in vitro, can be used for further development of wound-healing coatings. In addition, sponges made of chitosan glycolate can be used as highly effective hemostatic materials. Sponges made of chitosan ascorbate require further development and separate study.