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1.
NPJ Biofilms Microbiomes ; 10(1): 30, 2024 Mar 23.
Artigo em Inglês | MEDLINE | ID: mdl-38521769

RESUMO

Biofilms are surface-associated communities of bacteria that grow in a self-produced matrix of polysaccharides, proteins, and extracellular DNA (eDNA). Sub-minimal inhibitory concentrations (sub-MIC) of antibiotics induce biofilm formation, potentially as a defensive response to antibiotic stress. However, the mechanisms behind sub-MIC antibiotic-induced biofilm formation are unclear. We show that treatment of Pseudomonas aeruginosa with multiple classes of sub-MIC antibiotics with distinct targets induces biofilm formation. Further, addition of exogenous eDNA or cell lysate failed to increase biofilm formation to the same extent as antibiotics, suggesting that the release of cellular contents by antibiotic-driven bacteriolysis is insufficient. Using a genetic screen for stimulation-deficient mutants, we identified the outer membrane porin OprF and the ECF sigma factor SigX as important. Similarly, loss of OmpA - the Escherichia coli OprF homolog - prevented sub-MIC antibiotic stimulation of E. coli biofilms. Our screen also identified the periplasmic disulfide bond-forming enzyme DsbA and a predicted cyclic-di-GMP phosphodiesterase encoded by PA2200 as essential for biofilm stimulation. The phosphodiesterase activity of PA2200 is likely controlled by a disulfide bond in its regulatory domain, and folding of OprF is influenced by disulfide bond formation, connecting the mutant phenotypes. Addition of reducing agent dithiothreitol prevented sub-MIC antibiotic biofilm stimulation. Finally, activation of a c-di-GMP-responsive promoter follows treatment with sub-MIC antibiotics in the wild-type but not an oprF mutant. Together, these results show that antibiotic-induced biofilm formation is likely driven by a signaling pathway that translates changes in periplasmic redox state into elevated biofilm formation through increases in c-di-GMP.


Assuntos
Antibacterianos , Infecções por Pseudomonas , Humanos , Antibacterianos/farmacologia , Antibacterianos/metabolismo , Pseudomonas aeruginosa/fisiologia , Escherichia coli/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Biofilmes , Diester Fosfórico Hidrolases , Dissulfetos/metabolismo
2.
Helicobacter ; 29(2): e13064, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38459689

RESUMO

BACKGROUND: Helicobacter pylori (H. pylori) infection is the most extensively studied risk factor for gastric cancer. As with any bacteria, H. pylori will release distinctive odors that result from an emission of volatile metabolic byproducts in unique combinations and proportions. Effectively capturing and identifying these volatiles can pave the way for the development of innovative and non-invasive diagnostic methods for determining infection. Here we characterize the H. pylori volatilomic signature, pinpoint potential biomarkers of its presence, and evaluate the variability of volatilomic signatures between different H. pylori isolates. MATERIALS AND METHODS: Using needle trap extraction, volatiles in the headspace above H. pylori cultures were collected and, following thermal desorption at 290°C in a splitless mode, were analyzed using gas chromatography-mass spectrometry. The resulting volatilomic signatures of H. pylori cultures were compared to those obtained from an analysis of the volatiles in the headspace above the cultivating medium only. RESULTS: Amongst the volatiles detected, 21 showed consistent differences between the bacteria cultures and the cultivation medium, with 11 compounds being elevated and 10 showing decreased levels in the culture's headspace. The 11 elevated volatiles are four ketones (2-pentanone, 5-methyl-3-heptanone, 2-heptanone, and 2-nonanone), three alcohols (2-methyl-1-propanol, 3-methyl-1-butanol, and 1 butanol), one aromatic (styrene), one aldehyde (2-ethyl-hexanal), one hydrocarbon (n-octane), and one sulfur compound (dimethyl disulfide). The 10 volatiles with lower levels in the headspace of the cultures are four aldehydes (2-methylpropanal, benzaldehyde, 3-methylbutanal, and butanal), two heterocyclic compounds (2-ethylfuran and 2-pentylfuran), one ketone (2-butanone), one aromatic (benzene), one alcohol (2-butanol) and bromodichloromethane. Of the volatile species showing increased levels, the highest emissions are found to be for 3-methyl-1-butanol, 1-butanol and dimethyl disulfide. Qualitative variations in their emissions from the different isolates was observed. CONCLUSIONS: The volatiles emitted by H. pylori provide a characteristic volatilome signature that has the potential of being developed as a tool for monitoring infections caused by this pathogen. Furthermore, using the volatilome signature, we are able to differentiate different isolates of H. pylori. However, the volatiles also represent potential confounders for the recognition of gastric cancer volatile markers.


Assuntos
Dissulfetos , Infecções por Helicobacter , Helicobacter pylori , Pentanóis , Neoplasias Gástricas , Humanos , Álcoois
3.
Eur Rev Med Pharmacol Sci ; 28(4): 1471-1479, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38436181

RESUMO

OBJECTIVE: Thiols are organic compounds containing sulfhydryl groups that exert antioxidant effects via dynamic thiol-disulfide homeostasis. The shift towards disulfide indicates the presence of an oxidative environment. Different modes of delivery can affect thiol-disulfide homeostasis. Accordingly, we planned this research to evaluate the effects of the mode of delivery on thiol-disulfide homeostasis in both maternal serum and fetal cord blood samples. PATIENTS AND METHODS: We conducted a prospective case-control study involving two groups: vaginal delivery (n=50) and elective cesarean section (CS) (n=45). The vaginal delivery group exclusively comprised uncomplicated term deliveries, while the CS group included pregnant individuals with scheduled cesarean deliveries due to the absence of spontaneous labor onset. Maternal serum and fetal cord blood samples were collected, and thiol-disulfide exchanges were analyzed using an automated method capable of measuring both aspects of the thiol-disulfide balance. RESULTS: The levels of native thiol (-SH) and total thiol in both maternal serum and fetal cord blood samples were significantly higher in the vaginal delivery group than those in the CS group. An important discovery of our study was that fetal cord disulfide (-SS) level, which may reflect oxidative stress, was higher in newborns born via vaginal delivery when examined alone. However, in both maternal and fetal cord blood, the combined ratios, SS/SH ratio (%), SS/Total thiol ratio (%), and SH/Total thiol ratio (%) were observed to be similar between the groups in both maternal and fetal cord blood. It was observed that as the mother's weight gained during pregnancy increased, SS/SH and SS/total thiol increased (positive correlation), while SH/total thiol decreased (negative correlation). CONCLUSIONS: Our results showed that the dynamic thiol-disulfide homeostasis was greatly influenced by the way of delivery and supported the idea that vaginally-delivered infants may have more oxidative stress.


Assuntos
Cesárea , Parto Obstétrico , Recém-Nascido , Gravidez , Lactente , Humanos , Feminino , Estudos de Casos e Controles , Dissulfetos , Homeostase , Estresse Oxidativo , Compostos de Sulfidrila
4.
Protein Sci ; 33(4): e4949, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38511500

RESUMO

Primary defects in folding of mutant proinsulin can cause dominant-negative proinsulin accumulation in the endoplasmic reticulum (ER), impaired anterograde proinsulin trafficking, perturbed ER homeostasis, diminished insulin production, and ß-cell dysfunction. Conversely, if primary impairment of ER-to-Golgi trafficking (which also perturbs ER homeostasis) drives misfolding of nonmutant proinsulin-this might suggest bi-directional entry into a common pathological phenotype (proinsulin misfolding, perturbed ER homeostasis, and deficient ER export of proinsulin) that can culminate in diminished insulin storage and diabetes. Here, we've challenged ß-cells with conditions that impair ER-to-Golgi trafficking, and devised an accurate means to assess the relative abundance of distinct folded/misfolded forms of proinsulin using a novel nonreducing SDS-PAGE/immunoblotting protocol. We confirm abundant proinsulin misfolding upon introduction of a diabetogenic INS mutation, or in the islets of db/db mice. Whereas blockade of proinsulin trafficking in Golgi/post-Golgi compartments results in intracellular accumulation of properly-folded proinsulin (bearing native disulfide bonds), impairment of ER-to-Golgi trafficking (regardless whether such impairment is achieved by genetic or pharmacologic means) results in decreased native proinsulin with more misfolded proinsulin. Remarkably, reversible ER-to-Golgi transport defects (such as treatment with brefeldin A or cellular energy depletion) upon reversal quickly restore the ER folding environment, resulting in the disappearance of pre-existing misfolded proinsulin while preserving proinsulin bearing native disulfide bonds. Thus, proper homeostatic balance of ER-to-Golgi trafficking is linked to a more favorable proinsulin folding (as well as trafficking) outcome.


Assuntos
Diabetes Mellitus , Células Secretoras de Insulina , Camundongos , Animais , Proinsulina/genética , Proinsulina/química , Dobramento de Proteína , Insulina/química , Retículo Endoplasmático , Homeostase , Dissulfetos/química
5.
J Neuroinflammation ; 21(1): 70, 2024 Mar 21.
Artigo em Inglês | MEDLINE | ID: mdl-38515139

RESUMO

Myeloperoxidase (MPO) plays critical role in the pathology of cerebral ischemia-reperfusion (I/R) injury via producing hypochlorous acid (HOCl) and inducing oxidative modification of proteins. High-mobility group box 1 (HMGB1) oxidation, particularly disulfide HMGB1 formation, facilitates the secretion and release of HMGB1 and activates neuroinflammation, aggravating cerebral I/R injury. However, the cellular sources of MPO/HOCl in ischemic brain injury are unclear yet. Whether HOCl could promote HMGB1 secretion and release remains unknown. In the present study, we investigated the roles of microglia-derived MPO/HOCl in mediating HMGB1 translocation and secretion, and aggravating the brain damage and blood-brain barrier (BBB) disruption in cerebral I/R injury. In vitro, under the co-culture conditions with microglia BV cells but not the single culture conditions, oxygen-glucose deprivation/reoxygenation (OGD/R) significantly increased MPO/HOCl expression in PC12 cells. After the cells were exposed to OGD/R, MPO-containing exosomes derived from BV2 cells were released and transferred to PC12 cells, increasing MPO/HOCl in the PC12 cells. The HOCl promoted disulfide HMGB1 translocation and secretion and aggravated OGD/R-induced apoptosis. In vivo, SD rats were subjected to 2 h of middle cerebral artery occlusion (MCAO) plus different periods of reperfusion. Increased MPO/HOCl production was observed at the reperfusion stage, accomplished with enlarged infarct volume, aggravated BBB disruption and neurological dysfunctions. Treatment of MPO inhibitor 4-aminobenzoic acid hydrazide (4-ABAH) and HOCl scavenger taurine reversed those changes. HOCl was colocalized with cytoplasm transferred HMGB1, which was blocked by taurine in rat I/R-injured brain. We finally performed a clinical investigation and found that plasma HOCl concentration was positively correlated with infarct volume and neurological deficit scores in ischemic stroke patients. Taken together, we conclude that ischemia/hypoxia could activate microglia to release MPO-containing exosomes that transfer MPO to adjacent cells for HOCl production; Subsequently, the production of HOCl could mediate the translocation and secretion of disulfide HMGB1 that aggravates cerebral I/R injury. Furthermore, plasma HOCl level could be a novel biomarker for indexing brain damage in ischemic stroke patients.


Assuntos
Lesões Encefálicas , Isquemia Encefálica , Proteína HMGB1 , AVC Isquêmico , Traumatismo por Reperfusão , Humanos , Ratos , Animais , Ácido Hipocloroso , Microglia/metabolismo , Proteína HMGB1/metabolismo , Ratos Sprague-Dawley , Lesões Encefálicas/metabolismo , Isquemia Encefálica/metabolismo , Barreira Hematoencefálica/metabolismo , Infarto da Artéria Cerebral Média/complicações , Infarto da Artéria Cerebral Média/patologia , Neurônios/metabolismo , Traumatismo por Reperfusão/metabolismo , Peroxidase/metabolismo , Taurina , Dissulfetos
6.
ACS Sens ; 9(3): 1410-1418, 2024 Mar 22.
Artigo em Inglês | MEDLINE | ID: mdl-38456391

RESUMO

Dimethyl disulfide (DMDS) is a common odor pollutant with an extremely low olfactory threshold. Highly sensitive and selective detection of DMDS in ambient humid air background, by metal oxide semiconductor (MOS) sensors, is highly desirable to address the increased public concern for health risk. However, it has still been a critical challenge up to now. Herein, p-type delafossite CuGaO2 has been proposed as a promising DMDS sensing material owing to its striking hydrophobicity (revealed by water contact angle measurement) and excellent partial catalytic oxidation properties (indicated by mass spectroscopy). The present CuGaO2 sensor shows a selective DMDS response, with satisfied humidity resistance performance and long-term stability at a relatively low operation temperature of 140 °C. An ultrahigh response of 100 to 10 ppm DMDS and a low limit of detection of 3.3 ppb could be achieved via a pulsed temperature modulation strategy. A smart sensing system based on a CuGaO2 sensor has been developed, which could precisely monitor DMDS vapor in ambient humid air, even with the presence of multiple interfering gases, demonstrating the practical application capability of MOS sensors for environmental odor monitoring.


Assuntos
Dissulfetos , Gases , Óxidos/química , Temperatura
7.
ACS Nano ; 18(11): 7945-7958, 2024 Mar 19.
Artigo em Inglês | MEDLINE | ID: mdl-38452275

RESUMO

Ferroptosis is a form of regulated cell death accompanied by lipid reactive oxygen species (ROS) accumulation in an iron-dependent manner. However, the efficiency of tumorous ferroptosis was seriously restricted by intracellular ferroptosis defense systems, the glutathione peroxidase 4 (GPX4) system, and the ubiquinol (CoQH2) system. Inspired by the crucial role of mitochondria in the ferroptosis process, we reported a prodrug nanoassembly capable of unleashing potent mitochondrial lipid peroxidation and ferroptotic cell death. Dihydroorotate dehydrogenase (DHODH) inhibitor (QA) was combined with triphenylphosphonium moiety through a disulfide-containing linker to engineer well-defined nanoassemblies (QSSP) within a single-molecular framework. After being trapped in cancer cells, the acidic condition provoked the structural disassembly of QSSP to liberate free prodrug molecules. The mitochondrial membrane-potential-driven accumulation of the lipophilic cation prodrug was delivered explicitly into the mitochondria. Afterward, the thiol-disulfide exchange would occur accompanied by downregulation of reduced glutathione levels, thus resulting in mitochondria-localized GPX4 inactivation for ferroptosis. Simultaneously, the released QA from the hydrolysis reaction of the adjacent ester bond could further devastate mitochondrial defense and evoke robust ferroptosis via the DHODH-CoQH2 system. This subcellular targeted nanoassembly provides a reference for designing ferroptosis-based strategy for efficient cancer therapy through interfering antiferroptosis systems.


Assuntos
Ferroptose , Compostos Organofosforados , Pró-Fármacos , Pró-Fármacos/farmacologia , Pró-Fármacos/metabolismo , Di-Hidro-Orotato Desidrogenase , Peroxidação de Lipídeos , Espécies Reativas de Oxigênio/metabolismo , Mitocôndrias/metabolismo , Dissulfetos/metabolismo
8.
Metallomics ; 16(3)2024 Mar 12.
Artigo em Inglês | MEDLINE | ID: mdl-38425033

RESUMO

The tuberculosis (TB) emergency has been a pressing health threat for decades. With the emergence of drug-resistant TB and complications from the COVID-19 pandemic, the TB health crisis is more serious than ever. Mycobacterium tuberculosis (Mtb), the causative agent of TB, requires iron for its survival. Thus, Mtb has evolved several mechanisms to acquire iron from the host. Mtb produces two siderophores, mycobactin and carboxymycobactin, which scavenge for host iron. Mtb siderophore-dependent iron acquisition requires the export of apo-siderophores from the cytosol to the host environment and import of iron-bound siderophores. The export of Mtb apo-siderophores across the inner membrane is facilitated by two mycobacterial inner membrane proteins with their cognate periplasmic accessory proteins, designated MmpL4/MmpS4 and MmpL5/MmpS5. Notably, the Mtb MmpL4/MmpS4 and MmpL5/MmpS5 complexes have also been implicated in the efflux of anti-TB drugs. Herein, we solved the crystal structure of M. thermoresistibile MmpS5. The MmpS5 structure reveals a previously uncharacterized, biologically relevant disulfide bond that appears to be conserved across the Mycobacterium MmpS4/S5 homologs, and comparison with structural homologs suggests that MmpS5 may be dimeric.


Assuntos
Mycobacteriaceae , Mycobacterium tuberculosis , Tuberculose , Humanos , Pandemias , Mycobacterium tuberculosis/metabolismo , Tuberculose/microbiologia , Sideróforos/metabolismo , Ferro/metabolismo , Dissulfetos/metabolismo , Proteínas de Bactérias/metabolismo
9.
Mikrochim Acta ; 191(4): 190, 2024 03 09.
Artigo em Inglês | MEDLINE | ID: mdl-38460000

RESUMO

Golgi protein 73 (GP73) is a new serum marker associated with early diagnosis and postoperative assessment of hepatocellular carcinoma (HCC). Herein, an electrochemical/fluorescence dual-signal biosensor was designed for determination of GP73 based on molybdenum disulfide/ferrocene/palladium nanoparticles (MoS2-Fc-PdNPs) and nitrogen-doped graphene quantum dots (NGQDs). GP73 aptamer (Apt) was labeled with NGQDs to form the NGQDs-Apt fluorescence probe. MoS2-Fc-PdNPs served not only as the fluorescence quencher but also as electrochemical enhancer. The sensing platform (NGQDs-Apt/MoS2-Fc-PdNPs) was formed based on the fluorescence resonance energy transfer (FRET) mechanism. In the presence of GP73, the specific binding of NGQDs-Apt to GP73 interrupted FRET, restoring the fluorescence of NGQDs-Apt at λex/em = 348/438 nm and enhancing the oxidation current of Fc in MoS2-Fc-PdNPs at 0.04 V through differential pulse voltammetry (DPV). Under the optimal conditions, the DPV current change and fluorescence recovery have a good linear relationship with GP73 concentration from 1.00 to 10.0 ng/mL. The calibration equation for the fluorescence mode was Y1 = (0.0213 ± 0.00127)X + (0.0641 ± 0.00448) and LOD was 0.812 ng/mL (S/N = 3). The calibration equation of the electrochemical mode was Y2 = (3.41 ± 0.111)X + (1.62 ± 0.731), and LOD of 0.0425 ng/mL (S/N = 3). The RSDs of fluorescence mode and electrochemical mode after serum detection were 1.62 to 5.21% and 0.180 to 6.62%, respectively. By combining the electrochemical and fluorescence assay, more comprehensive and valuable information for GP73 was provided. Such dual-mode detection platform shows excellent reproducibility, stability, and selectivity and has great application potential.


Assuntos
Carcinoma Hepatocelular , Dissulfetos , Grafite , Neoplasias Hepáticas , Nanopartículas Metálicas , Pontos Quânticos , Humanos , Molibdênio , Paládio , Nitrogênio , Reprodutibilidade dos Testes , Metalocenos
10.
Methods Mol Biol ; 2778: 101-115, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38478274

RESUMO

Membrane-embedded ß-barrels are the major building blocks of the Gram-negative outer membrane and are involved in antibiotic resistance, virulence, and the maintenance of bacterial cell physiology. The increased frequency of multidrug resistant Gram-negative infections warrants the sharing of accessible methods for the study of ß-barrels. One such method is "in vivo disulfide-bond crosslinking" which is a highly informative and cost-effective approach to study the structure, topology, dynamicity, and function of ß-barrels in situ. The approach can also be used to identify and finely map both stable or transient interactions between ß-barrels and other interacting proteins. In this chapter, I describe the conceptual basis of in vivo disulfide-bond crosslinking and the potential pitfalls in experimental design. I also provide a general protocol for high-efficiency in vivo disulfide-bond crosslinking and modified protocols as examples for how the method can be adapted to different scenarios.


Assuntos
Proteínas da Membrana Bacteriana Externa , Dobramento de Proteína , Proteínas da Membrana Bacteriana Externa/metabolismo , Modelos Moleculares , Bactérias/metabolismo , Dissulfetos
11.
Int J Biol Macromol ; 263(Pt 2): 130431, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38403212

RESUMO

In this study, we propose a novel approach to improve the performance of chitosan coating, and thioctic acid with disulfide bonds in its molecular structure was grafted onto the side groups of chitosan macromolecules. The introduction of disulfide bond network cross-linking structure in chitosan coating weakens hydrogen bonds between chitosan macromolecules, causing the macromolecular chains to be more prone to relative motion when subjected to external forces, ultimately improving flexibility of the coating. The modified chitosan becomes more suitable for antibacterial modification in smart wearable fabrics. Subsequently, we fabricated a smart wearable fabric with excellent antibacterial properties and strong electromagnetic shielding by employing the layer-by-layer spraying technique. This involved incorporating chitosan with disulfide bonds and MXene nanoparticles. The fabric surfaces containing chitosan with disulfide bonds exhibited enhanced flexibility compared to unmodified chitosan fabric, resulting in an 8-point improvement in tactile sensation ratings. This research presents a novel approach that simultaneously enhances the electromagnetic shielding effectiveness and efficient antibacterial properties of smart wearable textiles. Consequently, it advances the application of chitosan in the field of antibacterial finishing for functional textiles.


Assuntos
Quitosana , Ácido Tióctico , Quitosana/química , Têxteis , Antibacterianos/farmacologia , Antibacterianos/química , Dissulfetos
12.
Int J Biol Macromol ; 262(Pt 2): 129953, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38325678

RESUMO

Tau cleavage has been shown to have a significant effect on protein aggregation. Tau truncation results in the formation of aggregation-prone fragments leading to toxic aggregates and also causes the formation of harmful fragments that do not aggregate. Thus, targeting proteolysis of tau would be beneficial for the development of therapeutics for Alzheimer's disease and related tauopathies. In this study, amino-terminal quantification and ThT fluorimetry were respectively used to analyze the kinetics of tau fragmentation and fibril formation. SDS-PAGE analysis of tau protein incubated with a disulfide-reducing agent demonstrated that the cysteines of tau have a crucial role in the fibrillation and autoproteolysis. However, the structures converted to amyloid fibrils were different with conformations that led to autoproteolysis. The quantification of the amino terminal indicated that the double-disulfide parallel structures formed in the presence of heparin did not have protease activity. The survey of possible tau disulfide-mediated dimer configurations suggested that the non-register single disulfide bound conformations were involved in the tau autoproteolysis process. Moreover, the inhibition of autoproteolysis resulted in the increment of aggregation rate; hence it seems that the tau auto-cleavage is the cellular defense mechanism against protein fibrillation.


Assuntos
Doença de Alzheimer , Tauopatias , Humanos , Proteínas tau/química , Amiloide/química , Doença de Alzheimer/metabolismo , Tauopatias/metabolismo , Dissulfetos
13.
Int J Biol Macromol ; 262(Pt 2): 130222, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38365145

RESUMO

Understanding the mechanism by which Triticeae improves the quality of broomcorn millet (BM) is key to expanding the use of this crop to address food crises and food security. This study aimed to explore the effects of Triticeae on the disulfide bonds, secondary structures, microstructure, and rheological properties of BM dough, and to investigate the potential food applications of BM. Gluten protein, intermolecular SS, and ß-Sheets content of the reconstituted doughs were significantly improved compared with BM dough, which improved disorderly accumulation of starch and gluten-starch interaction in BM dough. CLSM analysis showed that broomcorn millet-common wheat (BM-CW) and broomcorn millet-durum wheat (BM-DW) also possessed larger protein areas, smaller lacunarities, and better gluten-starch interactions in the reconstituted doughs. Disulfide bonds were positively correlated with the gluten network structure, and more disulfide bonds were formed in BM-CW (3.86 µmol/g), which promoted stronger mechanical resistance in BM-CW. Therefore, the combination of BM flour with CW and DW flours had better dough elasticity and stability. Finally, a potential evaluation and optimization scheme for BM as a cooked wheaten food is proposed to improve the reference for future food security and dietary structure adjustment of residents.


Assuntos
Panicum , Amido , Amido/química , Glutens/química , Panicum/química , Triticum/química , Dissulfetos , Farinha
14.
Anim Sci J ; 95(1): e13917, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38323750

RESUMO

Allicin is a sulfur-containing compound extracted from raw garlic (Allium sativum L.). We compared the effect of allicin addition on growth performance, serum biochemical parameters, and rumen microbiota of goats compared to monensin. Twenty-four Anhui white goats were assigned randomly to one of three dietary treatments: 1) a basal diet (CON); 2) the basal diet with allicin addition at 750 mg per head per day (AC); 3) the basal diet with monensin addition at 30 mg per kg of diet (MS). Animals were fed for 8 weeks. Results showed the average daily gain, and feed efficiency was increased with allicin and monensin addition. Serum levels of IgG, total superoxide dismutase, and glutathione peroxidase were higher in the AC group than those in the CON and MS groups. The microbiota analysis revealed that monensin addition mainly affected genera related to carbohydrate and protein metabolism, and allicin mainly affected genera related to energy metabolism and intestinal health. In conclusion, allicin could improve growth performance and have advantages over monensin in improving the antioxidant capacity and immune function of goats. Allicin may be a potential alternative to monensin.


Assuntos
Dissulfetos , Alho , Microbiota , Ácidos Sulfínicos , Animais , Ração Animal/análise , Antioxidantes/metabolismo , Dieta/veterinária , Suplementos Nutricionais/análise , Cabras/metabolismo , Monensin/farmacologia , Rúmen/metabolismo
15.
Int J Biol Sci ; 20(3): 1042-1044, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38322120

RESUMO

Disulfidptosis occurs as a result of the accumulation of intracellular cystine followed by disulfide stress in actin cytoskeleton proteins due to a reduction of NADPH produced through the pentose phosphate pathway in cells with high expression of SLC7A11. It is a cell death caused by the redox imbalance resulting from the disruption of amino acid metabolism and glucose metabolism. The discovery of disulfidptosis has sparked immense enthusiasm, but there are numerous unresolved issues that need to be addressed. Solutions to these riddles will provide insights into the detailed mechanisms and the pathophysiological relevance of disulfidptosis and utilizing disulfidptosis as an actionable therapeutic target.


Assuntos
Dissulfetos , Proteínas dos Microfilamentos , Morte Celular , NADP
16.
Microb Cell Fact ; 23(1): 48, 2024 Feb 12.
Artigo em Inglês | MEDLINE | ID: mdl-38347541

RESUMO

BACKGROUND: The three-finger proteins are a collection of disulfide bond rich proteins of great biomedical interests. Scalable recombinant expression and purification of bioactive three-finger proteins is quite difficult. RESULTS: We introduce a working pipeline for expression, purification and validation of disulfide-bond rich three-finger proteins using E. coli as the expression host. With this pipeline, we have successfully obtained highly purified and bioactive recombinant α-Βungarotoxin, k-Bungarotoxin, Hannalgesin, Mambalgin-1, α-Cobratoxin, MTα, Slurp1, Pate B etc. Milligrams to hundreds of milligrams of recombinant three finger proteins were obtained within weeks in the lab. The recombinant proteins showed specificity in binding assay and six of them were crystallized and structurally validated using X-ray diffraction protein crystallography. CONCLUSIONS: Our pipeline allows refolding and purifying recombinant three finger proteins under optimized conditions and can be scaled up for massive production of three finger proteins. As many three finger proteins have attractive therapeutic or research interests and due to the extremely high quality of the recombinant three finger proteins we obtained, our method provides a competitive alternative to either their native counterparts or chemically synthetic ones and should facilitate related research and applications.


Assuntos
Escherichia coli , Corpos de Inclusão , Escherichia coli/metabolismo , Proteínas Recombinantes , Corpos de Inclusão/metabolismo , Dissulfetos/metabolismo
17.
ACS Appl Mater Interfaces ; 16(6): 6859-6867, 2024 Feb 14.
Artigo em Inglês | MEDLINE | ID: mdl-38299497

RESUMO

The rapid development of nanomedicine has considerably advanced precision therapy for cancer treatment. Superior to traditional chemotherapy, emerging theranostic nanoprodrugs can effectively realize inherent self-tracking, targeted drug delivery, stimuli-triggered drug release, and reduced systemic toxicity of chemotherapeutic drugs. However, theranostic nanoprodrugs with real-time drug release monitoring have remained rare so far. In this work, we developed a new glutathione-responsive theranostic nanoprodrug with a high drug-loading content of 59.4 wt % and an average nanoscale size of 46 nm, consisting of the anticancer drug paclitaxel and a fluorescent imaging probe with a high fluorescence quantum yield, which are linked by a disulfide-based glutathione-sensitive self-immolating linker. The strong fluorescence emission of the fluorophore enables efficacious self-tracking and sensitive fluorescence "ON-OFF" glutathione sensing. Upon encountering high-level glutathione in cancer cells, the disulfide bond is cleaved, and the resulting linker halves spontaneously collapse into cyclic small molecules at the same pace, leading to the simultaneous release of the therapeutic drug and the fluorescence-OFF imaging probe. Thereby, the drug release process is efficiently monitored by the fluorescence change in the nanoprodrug. The nanoprodrugs exerted high cytotoxicity toward various cancer cells, especially for A549 and HEK-293 cells, in which the nanoprodrugs generated better therapeutic effects than free paclitaxel. Our work demonstrated a new modality of smart theranostic nanoprodrugs for precise cancer therapy.


Assuntos
Nanopartículas , Neoplasias , Humanos , Medicina de Precisão , Linhagem Celular Tumoral , Liberação Controlada de Fármacos , Elétrons , Células HEK293 , Paclitaxel/farmacologia , Paclitaxel/uso terapêutico , Nanomedicina Teranóstica/métodos , Imagem Óptica/métodos , Glutationa/metabolismo , Dissulfetos/uso terapêutico , Nanopartículas/química , Neoplasias/diagnóstico por imagem , Neoplasias/tratamento farmacológico
18.
Behav Brain Res ; 462: 114894, 2024 Mar 28.
Artigo em Inglês | MEDLINE | ID: mdl-38311071

RESUMO

This study investigates the impact of orlistat on oxidative stress, spatial memory, recognition memory, and hippocampal tissue in obese rats. The study groups were divided into control, high fat diet-induced obese (HFDIO), HFDIO+orlistat (HFDIO+ORL) groups, each consisting of 8 animals. While control fed with standart diet, HFDIO and HFDIO+ORL fed with high-fat diets for 8 weeks to induce obesity. Then, ORL treated 10 mg/kg for 7 weeks, while control and HFDIO get water. At 16th week, novel object recognition (NOR) and Morris water maze (MWM) tests were performed. TNF-alpha, IL-1beta levels in hippocampal tissue, and total/native thiol/disulphide levels in serum were measured. TNF-alpha level of HFDIO was higher than control, while lower in HFDIO+ORL compared to HFDIO as like IL-1beta level. On the contrary, serum total thiol level was lower in HFDIO than control and higher in HFDIO+ORL compared to the HFDIO, while disulphide level was opposite of the total thiol levels. While recognition index was higher in HFDIO+ORL, in MWM, latency of finding platform in HFDIO was higher than control and latency of HFDIO+ORL was very similar to control in 2-4 days. The HFDIO group demonstrated decrease in time spent in platform zone compared to control, whereas time spent of the HFDIO+ORL was higher than HFDIO. Our study demonstrates that orlistat administration exerts beneficial effects on oxidative stress, spatial memory, recognition memory, and hippocampal tissue in obese rats. It shows that orlistat may have potential therapeutic implications for obesity-related cognitive impairments and hippocampal dysfunction.


Assuntos
Memória Espacial , Fator de Necrose Tumoral alfa , Ratos , Animais , Orlistate/farmacologia , Fator de Necrose Tumoral alfa/farmacologia , Hipocampo , Estresse Oxidativo , Obesidade/tratamento farmacológico , Dieta Hiperlipídica/efeitos adversos , Dissulfetos/farmacologia , Compostos de Sulfidrila/farmacologia
19.
Rapid Commun Mass Spectrom ; 38(7): e9713, 2024 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-38361473

RESUMO

RATIONALE: Disulfide bridges (DSB) play an important role in stabilizing three-dimensional structures of biopharmaceuticals, single purified proteins, and various cyclic peptide drugs that contain disulfide in their structures. Incorrect cross-linking known as DSB scrambling results in misfolded structures that can be inactive, immunogenic, and susceptible to aggregation. Very few articles have been published on the experimental annotation of DSBs in proteins and cyclic peptide drugs. Accurate characterization of the disulfide bond is essential for understanding protein confirmation. METHODS: Characterizing DSBs using mass spectrometry (MS) involves the chemical and enzymatic digestion of samples to obtain smaller peptide fragments, in both reduced and nonreduced forms. Subsequently, these samples are analyzed using MS to locate the DSB, either through interpretation or by employing various software tools. RESULTS: The main challenge in DSB analysis methods using sample preparation is to obtain a sample solution in which nonnative DSBs are not formed due to high pH, temperature, and presence of free sulfhydryl groups. Formation of nonnative DSBs can lead to erroneous annotation of disulfide bond. Sample preparation techniques, fragmentation methods for DSB analysis, and contemporary approaches for DSB mapping using this fragmentation were discussed. CONCLUSIONS: This review presents the latest advancement in MS-based characterization; also a critical perspective is presented for further annotation of DSBs using MS, primarily for single purified proteins or peptides that are densely connected and rich in cysteine. Despite significant breakthroughs resulting from advancements in MS, the analysis of disulfide bonds is not straightforward; it necessitates expertise in sample preparation and interpretation.


Assuntos
Peptídeos , Proteínas , Espectrometria de Massas , Proteínas/química , Peptídeos/análise , Peptídeos Cíclicos , Dissulfetos/química
20.
J Am Chem Soc ; 146(6): 3974-3983, 2024 Feb 14.
Artigo em Inglês | MEDLINE | ID: mdl-38299512

RESUMO

Biologics, including proteins and antisense oligonucleotides (ASOs), face significant challenges when it comes to achieving intracellular delivery within specific organs or cells through systemic administrations. In this study, we present a novel approach for delivering proteins and ASOs to liver cells, both in vitro and in vivo, using conjugates that tether N-acetylated galactosamine (GalNAc)-functionalized, cell-penetrating polydisulfides (PDSs). The method involves the thiol-bearing cargo-mediated ring-opening polymerization of GalNAc-functionalized lipoamide monomers through the so-called aggregation-induced polymerization, leading to the formation of site-specific protein/ASO-PDS conjugates with narrow dispersity. The hepatocyte-selective intracellular delivery of the conjugates arises from a combination of factors, including first GalNAc binding with ASGPR receptors on liver cells, leading to cell immobilization, and the subsequent thiol-disulfide exchange occurring on the cell surface, promoting internalization. Our findings emphasize the critical role of the close proximity of the PDS backbone to the cell surface, as it governs the success of thiol-disulfide exchange and, consequently, cell penetration. These conjugates hold tremendous potential in overcoming the various biological barriers encountered during systemic and cell-specific delivery of biomacromolecular cargos, opening up new avenues for the diagnosis and treatment of a range of liver-targeting diseases.


Assuntos
Produtos Biológicos , Galactosamina , Galactosamina/química , Hepatócitos/metabolismo , Oligonucleotídeos Antissenso/química , Dissulfetos/metabolismo , Compostos de Sulfidrila/metabolismo , Produtos Biológicos/metabolismo
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