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1.
Reprod Domest Anim ; 59(5): e14569, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38715435

RESUMO

The effects of an aqueous extract of Scabiosa atropurpurea L. (AES) on the reproduction potential of Queue Fine de l'Ouest rams were evaluated over 9 weeks. Eighteen mature (4-6 years old) rams (52.8 ± 2.6 kg) were divided into three groups. The control (C) group was fed oat hay ad libitum with 700 g of concentrate and the other two groups were fed the same diet supplemented with AES at 1 and 2 mg/kg body weight (AES1 and AES2, respectively). Ram sperm was collected with an artificial vagina (2 × 2 days/week) to evaluate sperm production and quality, antioxidant activity, the adenosine triphosphate (ATP) and calcium concentrations. Sexual behaviour and plasma testosterone concentrations were also investigated. The administration of AES improved sexual behaviour (the duration of contact and the number of lateral approaches). The addition of AES also improved individual spermatozoa motility (C: 71.7% ± 6.3%; AES1: 78.3% ± 4.9%; AES2: 83.8% ± 4.4%), the sperm concentration (C: 5.6 ± 0.36; AES1: 6.4 ± 0.81; AES2: 6.7 ± 0.52 × 109 spermatozoa/mL), the ATP ratio (C: 1 ± 0.08; AES1: 2.1 ± 0.08; AES2: 3.3 ± 0.08) and the calcium concentration (C: 5.6 ± 0.24; AES1: 7.7 ± 0.21; AES2: 8.1 ± 0.24 mmol/L). AES treatment decreased the percentage of abnormal sperm (C: 18.5% ± 1.2%; AES1: 16.2% ± 1.1%; AES2: 14.8% ± 0.94%) and DNA damage (C: 62%; AES1: 27%; AES2: 33%) and was associated with elevated seminal fluid antioxidant activity (C: 22 ± 0.27; AES1: 27.1 ± 1.08 and AES2: 27.5 ± 0.36 mmol Trolox equivalents/L) and plasma testosterone (C: 8.3 ± 0.7; AES1: 11.7 ± 0.4; AES2: 15 ± 0.7 ng/L). In conclusion, our study suggests that S. atropurpurea may be potentially useful to enhance libido and sperm production and quality in ram.


Assuntos
Extratos Vegetais , Comportamento Sexual Animal , Espermatozoides , Masculino , Animais , Espermatozoides/efeitos dos fármacos , Comportamento Sexual Animal/efeitos dos fármacos , Extratos Vegetais/farmacologia , Extratos Vegetais/administração & dosagem , Testosterona/sangue , Análise do Sêmen/veterinária , Motilidade dos Espermatozoides/efeitos dos fármacos , Suplementos Nutricionais , Antioxidantes/farmacologia , Dieta/veterinária , Contagem de Espermatozoides , Cálcio/análise , Cálcio/sangue , Carneiro Doméstico , Trifosfato de Adenosina/metabolismo , Trifosfato de Adenosina/análise
2.
Analyst ; 149(10): 2796-2800, 2024 May 13.
Artigo em Inglês | MEDLINE | ID: mdl-38669149

RESUMO

A near-infrared fluorescent nanoprobe consisting of Nile blue-capped ZIF-90 is first proposed for real-time imaging of mitochondrial ATP. Owing to the strong binding of ATP with Zn2+, the structure of the probe is disrupted, leading to the release of fluorescent NB.


Assuntos
Trifosfato de Adenosina , Corantes Fluorescentes , Mitocôndrias , Oxazinas , Corantes Fluorescentes/química , Corantes Fluorescentes/síntese química , Oxazinas/química , Humanos , Mitocôndrias/química , Mitocôndrias/metabolismo , Trifosfato de Adenosina/análise , Trifosfato de Adenosina/química , Trifosfato de Adenosina/metabolismo , Células HeLa , Raios Infravermelhos , Imagem Óptica/métodos , Nanopartículas/química
3.
Anal Chem ; 96(16): 6202-6208, 2024 Apr 23.
Artigo em Inglês | MEDLINE | ID: mdl-38598750

RESUMO

New strategies for accurate and reliable detection of adenosine triphosphate (ATP) with portable devices are significant for biochemical analysis, while most recently reported approaches cannot satisfy the detection accuracy and independent of large instruments simultaneously, which are unsuitable for fast, simple, and on-site ATP monitoring. Herein, a unique, convenient, and label-free point-of-care sensing strategy based on novel copper coordination polymer nanoflowers (CuCPNFs) was fabricated for multimode (UV-vis, photothermal, and RGB values) onsite ATP determination with high selectivity, sensitivity, and accuracy. The resulting CuCPNFs with a 3D hierarchical structure exhibit the ATP-triggered decomposition behavior because the competitive coordination between ATP and the copper ions of CuCPNFs can result in the formation of ATP-Cu, which reveals preeminent peroxidase mimics activity and can accelerate the oxidation of 3, 3', 5, 5'-tetramethylbenzidine (TMB) to form oxTMB. During this process, the detection system displayed not only color changes but also a strong NIR laser-driven photothermal effect. Thus, the photothermal and color signal variations are easily monitored by a portable thermometer and a smartphone. This multimode point-of-care platform can meet the requirements of onsite, without bulky equipment, accuracy, and reliability all at once, greatly enhancing its application in practice and paving a new way in ATP analysis.


Assuntos
Trifosfato de Adenosina , Cobre , Polímeros , Cobre/química , Trifosfato de Adenosina/análise , Polímeros/química , Sistemas Automatizados de Assistência Junto ao Leito , Humanos , Nanoestruturas/química , Limite de Detecção , Colorimetria , Benzidinas/química , Testes Imediatos
4.
Biosens Bioelectron ; 257: 116334, 2024 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-38678788

RESUMO

Burkholderia pseudomallei, widely distributed in tropical and subtropical ecosystems, is capable of causing the fatal zoonotic disease melioidosis and exhibiting a global trend of dissemination. Rapid and sensitive detection of B. pseudomallei is essential for environmental monitoring as well as infection control. Here, we developed an innovative biosensor for quantitatively detecting B. pseudomallei relies on ATP released triggered by bacteriophage-induced bacteria lysis. The lytic bacteriophage vB_BpP_HN01, with high specificity, is employed alongside magnetic nanoparticles assembly to create a biological receptor, facilitating the capture and enrichment of viable target bacteria. Following a brief extraction and incubation process, the captured target undergoes rapid lysis to release contents including ATP. The EXPAR-CRISPR cascade reaction provides an efficient signal transduction and dual amplification module that allowing the generated ATP to guide the signal output as an activator, ultimately converting the target bacterial amount into a detectable fluorescence signal. The proposed bacteriophage affinity strategy exhibited superior performance for B. pseudomallei detection with a dynamic range from 10^2 to 10^7 CFU mL-1, and a LOD of 45 CFU mL-1 within 80 min. Moreover, with the output signal compatible across various monitoring methods, this work offers a robust assurance for rapid diagnosis and on-site environmental monitoring of B. pseudomallei.


Assuntos
Trifosfato de Adenosina , Bacteriófagos , Técnicas Biossensoriais , Burkholderia pseudomallei , Sistemas CRISPR-Cas , Burkholderia pseudomallei/virologia , Técnicas Biossensoriais/métodos , Bacteriófagos/química , Bacteriófagos/isolamento & purificação , Trifosfato de Adenosina/metabolismo , Trifosfato de Adenosina/análise , Melioidose/microbiologia , Limite de Detecção , Humanos , Nanopartículas de Magnetita/química
5.
Talanta ; 274: 126025, 2024 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-38574539

RESUMO

Exposure to bioaerosol contamination has detrimental effects on human health. Recent advances in ATP bioluminescence provide more opportunities for the quantitative detection of bioaerosols. Since almost all active organisms can produce ATP, the amount of airborne microbes can be easily measured by detecting ATP-driven bioluminescence. The accurate evaluation of microorganisms mainly relies on following the four key steps: sampling and enrichment of airborne microbes, lysis for ATP extraction, enzymatic reaction, and measurement of luminescence intensity. To enhance the effectiveness of ATP bioluminescence, each step requires innovative strategies and continuous improvement. In this review, we summarized the recent advances in the quantitative detection of airborne microbes based on ATP bioluminescence, which focuses on the advanced strategies for improving sampling devices combined with ATP bioluminescence. Meanwhile, the optimized and innovative strategies for the remaining three key steps of the ATP bioluminescence assay are highlighted. The aim is to reawaken the prosperity of ATP bioluminescence and promote its wider utilization for efficient, real-time, and accurate detection of airborne microbes.


Assuntos
Trifosfato de Adenosina , Microbiologia do Ar , Medições Luminescentes , Trifosfato de Adenosina/análise , Medições Luminescentes/métodos , Bactérias/isolamento & purificação , Humanos , Monitoramento Ambiental/métodos
6.
Diagn Microbiol Infect Dis ; 109(2): 116275, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38537505

RESUMO

BACKGROUND: Tuberculosis (TB), caused by Mycobacterium tuberculosis (MTB), remains a global health threat, necessitating faster and more accessible diagnostic methods. This study investigates critical parameters in the application of a commercial ATP bioluminescence assay for the detection of MTB. METHOD: Our objective was to optimize the ATP bioluminescence protocol using BacTiter-Glo™ for MTB, investigating the impact of varying volumes of MTB suspension and reagent on assay sensitivity, evaluating ATP extraction methods, establishing calibration curves, and elucidating strain-specific responses to antimicrobial agents. RESULTS: ATP extraction methods showed no significant improvement over controls. Calibration curves revealed a linear correlation between relative light units (RLU) and colony-forming units (CFU/mL), establishing low detection limits. Antimicrobial testing demonstrated strain-specific responses aligning with susceptibility and resistance patterns. CONCLUSION: Our findings contribute to refining ATP bioluminescence protocols for enhanced MTB detection and susceptibility testing. Further refinements and validation efforts are warranted, holding promise for more efficient diagnostic platforms in the future.


Assuntos
Trifosfato de Adenosina , Medições Luminescentes , Mycobacterium tuberculosis , Tuberculose , Mycobacterium tuberculosis/efeitos dos fármacos , Trifosfato de Adenosina/análise , Trifosfato de Adenosina/metabolismo , Medições Luminescentes/métodos , Humanos , Tuberculose/diagnóstico , Tuberculose/microbiologia , Sensibilidade e Especificidade , Testes de Sensibilidade Microbiana/métodos , Técnicas Bacteriológicas/métodos
7.
Talanta ; 274: 125992, 2024 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-38552479

RESUMO

Organic photoelectrochemical transistor (OPECT) biosensor is now appearing in perspective of public, which characterized by amplified the grating electrode potential by ion transport. In this study, the DNA network formed by the hybridization chain reaction (HCR) detects the target adenosine triphosphate (ATP) by adjusting the surface potential of the new heterojunction of ZnIn2S4/MXene. The formation of DNA network amplifies the detection signal of ATP. Significantly, OPECT biosensor could further amplify the signal, which calculated the gain achieved 103, which is consistent with the gain signal of the previously reported OPECT biosensor. Furthermore, the OPECT biosensor achieved a highly sensitivity detection of the target ATP, which the linear detection range is 0.03 pM-30 nM, and the detection limit is 0.03 pM, and illustrated a high selectivity to ATP. The proposed OPECT biosensor achieved signal amplification by adjusting the surface potential of ZnIn2S4/MXene through cascade DNA network, which provides a new direction for the detection of biomolecules.


Assuntos
Trifosfato de Adenosina , Técnicas Biossensoriais , DNA , Técnicas Eletroquímicas , Transistores Eletrônicos , Zinco , Trifosfato de Adenosina/análise , Trifosfato de Adenosina/química , Técnicas Biossensoriais/métodos , DNA/química , DNA/análise , Técnicas Eletroquímicas/métodos , Técnicas Eletroquímicas/instrumentação , Zinco/química , Índio/química , Processos Fotoquímicos , Limite de Detecção , Hibridização de Ácido Nucleico
8.
ACS Appl Mater Interfaces ; 16(4): 5129-5137, 2024 Jan 31.
Artigo em Inglês | MEDLINE | ID: mdl-38227932

RESUMO

Adenosine triphosphate (ATP) is a small molecule that is released to the urine from bladder urothelial cells and the bladder mucosal band of the human body. In certain cases, ATP can serve as a biomarker in bladder disease. For the practical applicability of luminescent sensors for ATP in urine, it is significant to find a new strategy for making the detection progress simple and available for in-field urine analysis. Here, a novel luminescent lanthanide coordination polymer (Tb-BPA) was designed and synthesized for quick and sensitive detection of ATP through luminescence quenching with a quenching constant of 4.90 × 103 M-1 and a detection limit of 0.55 × 10-6 M. Besides, Tb-BPA has excellent anti-interference ability and can detect ATP in simulated urine with a small relative standard deviation (<4%). Moreover, the luminescent polyacrylonitrile nanofiber films modified by Tb-BPA were prepared by electrospinning and were used for ATP visual detection. Notably, this film is easy to recover and reuse, and maintains good detection performance after at least 7 cycles.


Assuntos
Elementos da Série dos Lantanídeos , Humanos , Trifosfato de Adenosina/análise , Polímeros , Luminescência
9.
J Sci Food Agric ; 104(3): 1458-1469, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-37814322

RESUMO

BACKGROUND: Streptococcus thermophilus is an important strain widely used in dairy fermentation, with distinct urea metabolism characteristics compared to other lactic acid bacteria. The conversion of urea by S. thermophilus has been shown to affect the flavor and acidification characteristics of milk. Additionally, urea metabolism has been found to significantly increase the number of cells and reduce cell damage under acidic pH conditions, resulting in higher activity. However, the physiological role of urea metabolism in S. thermophilus has not been fully evaluated. A deep understanding of this metabolic feature is of great significance for its production and application. Genome-scale metabolic network models (GEMs) are effective tools for investigating the metabolic network of organisms using computational biology methods. Constructing an organism-specific GEM can assist us in comprehending its characteristic metabolism at a systemic level. RESULTS: In the present study, we reconstructed a high-quality GEM of S. thermophilus S-3 (iCH492), which contains 492 genes, 608 metabolites and 642 reactions. Growth phenotyping experiments were employed to validate the model both qualitatively and quantitatively, yielding satisfactory predictive accuracy (95.83%), sensitivity (93.33%) and specificity (100%). Subsequently, a systematic evaluation of urea metabolism in S. thermophilus was performed using iCH492. The results showed that urea metabolism reduces intracellular hydrogen ions and creates membrane potential by producing and transporting ammonium ions. This activation of glycolytic fluxes and ATP synthase produces more ATP for biomass synthesis. The regulation of fluxes of reactions involving NAD(P)H by urea metabolism improves redox balance. CONCLUSION: Model iCH492 represents the most comprehensive knowledge-base of S. thermophilus to date, serving as a potent tool. The evaluation of urea metabolism led to novel insights regarding the role of urease. © 2023 Society of Chemical Industry.


Assuntos
Redes e Vias Metabólicas , Streptococcus thermophilus , Animais , Streptococcus thermophilus/genética , Streptococcus thermophilus/metabolismo , Fermentação , Leite/química , Ureia/metabolismo , Trifosfato de Adenosina/análise
10.
Biosens Bioelectron ; 241: 115691, 2023 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-37738803

RESUMO

For discriminating diverse analytes and monitoring a specific chemical reaction, the emerging multi-channel "chemical nose/tongue" is challenging multi-material "chemical nose/tongue". The former contributes greatly to integrating different transduction principles from a single sensing material, avoiding the need for complex design, high cost, and tedious operation involved with the latter. Therefore, this high-order sensing puts a particular emphasis on the effects of encapsulation. Herein, the plasmonic gold nanoparticles (Au NPs) are encapsulated as a core into the fluorescent guanine monophosphate-Tb3+ infinite coordination polymer nanoparticles (GMP-Tb ICPs) to obtain a core-shell nanocomposite named Au NPs@GMP-Tb ICPs. Hence, a dual-channel "chemical tongue" based on Au NPs@GMP-Tb ICPs is present to realize high-order sensing of adenosine triphosphate (ATP)-related physiological phosphates and the monitoring of ATP hydrolysis. Considering the affinity of Tb3+ towards P-O bonds, four inorganic phosphates and three nucleotide phosphates with different phosphate group numbers and steric hindrance effect directly regulate two stimulus responses (fluorescence intensity and UV-vis absorbance) of Au NPs@GMP-Tb ICPs. Robust statistical methods, such as linear discriminant analysis and hierarchical cluster analysis, are used to recognize each phosphate by the developed sensor array either in the aqueous solution or in complex media such as serum, together with efficiently monitored ATP hydrolysis at different intervals. These findings and blind test clarify that the designed "chemical tongue" guarantees interference resistance and strengthens analytical capacity, together with offering valuable insight into "lab-on-a-nanoparticle" development for monitoring specific chemical reactions.


Assuntos
Técnicas Biossensoriais , Nanopartículas Metálicas , Trifosfato de Adenosina/análise , Ouro/química , Hidrólise , Nanopartículas Metálicas/química , Técnicas Biossensoriais/métodos , Fosfatos
11.
J Osteopath Med ; 123(12): 585-591, 2023 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-37698520

RESUMO

CONTEXT: Healthcare-associated infections (HAIs) pose a substantial public health threat. Despite significant strides to curb HAIs in hospital environments, outpatient settings have not received the same degree of attention. Given their emphasis on holistic, patient-centered care, osteopathic family medicine offices are pivotal in both disease prevention and comprehensive patient treatment. The importance of simple yet effective disinfection protocols, such as thorough cleaning between patient appointments, cannot be overstated in these settings because they are integral to minimizing disease transmission. OBJECTIVES: This study aims to assess the effectiveness of the current disinfection protocols in osteopathic family medicine offices. METHODS: A cross-sectional study evaluating disinfection practices on 18 examination tables in an osteopathic family medicine office was conducted. Two high-touch surfaces (midtorso region and table edge) were examined. Initial swab samples were collected after morning disinfection by Environmental Services, and terminal swab samples were gathered after day's-end disinfection by the medical staff. Adenosine triphosphate (ATP) bioluminescence assays were performed utilizing AccuPoint Advanced HC Reader, which quantified ATP, indicating contamination levels in the samples. The higher the ATP levels found in a sample, the greater the amount of biological contamination. All samplers were handled and tested as per manufacturer's instructions. A preliminary trial was conducted to confirm the internal validity of ATP bioluminescence measurements. The statistical analysis involved Shapiro-Wilk and Wilcoxon signed-rank tests, with significance set at p<0.05. Cohen's d test was utilized to calculate the effect size, identifying meaningful differences in initial and terminal swab sample relative light units (RLUs). RESULTS: The midtorso region demonstrated an 11.1 % increase in failure rate after terminal disinfection when compared to initial disinfection. A Wilcoxon signed-rank test revealed a median estimated pathogen level for the midtorso region that was higher after terminal disinfection (median, 193 RLUs; range, 1-690 RLUs; n=18) compared to initial disinfection (median, 134 RLUs; range, 4-946 RLUs; n=18). However, this increase was not statistically significant, p=0.9124, with a small effect size, d=0.04. The edge showed no change in failure rate after terminal disinfection, maintaining a 100 % failure rate both before and after disinfection. However, the Wilcoxon signed-rank test revealed a slight reduction in the median estimated pathogen levels after terminal disinfection (median, 2095 RLUs; range, 891-5,540 RLUs; n=18) compared to before disinfection (median, 2,257 RLUs; range, 932-5,825 RLUs; n=18). However, this reduction was not statistically significant, p=0.61, with a small effect size, d=0.12. CONCLUSIONS: The findings from this study reveal a substantial disparity in outcomes between the two sample locations, midtorso and edge. The midtorso demonstrated a relatively low failure rate in both initial and terminal swab samples, indicating successful outcomes. In contrast, the edge consistently displayed a 100 % failure rate, emphasizing the need for more care and attention when cleaning the edge of the examination to ensure better outcomes. By prioritizing adequate disinfection protocols, including thorough cleaning between patients, osteopathic family medicine offices can more effectively prevent disease transmission and promote patient safety.


Assuntos
Infecção Hospitalar , Desinfecção , Humanos , Desinfecção/métodos , Estudos Transversais , Medicina de Família e Comunidade , Infecção Hospitalar/prevenção & controle , Trifosfato de Adenosina/análise
12.
Biosens Bioelectron ; 234: 115356, 2023 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-37172362

RESUMO

The COVID-19 pandemic ignited massive research into the rapid detection of bioaerosols. In particular, nanotechnology-based detection strategies are proposed as alternatives because of issues in bioaerosol enrichment and lead time for molecular diagnostics; however, the practical implementation of such techniques is still unclear due to obstacles regarding the large research and development effort and investment for the validation. The use of adenosine triphosphate (ATP) bioluminescence (expressed as relative luminescence unit (RLU) per unit volume of air) of airborne particulate matter (PM) to determine the bacterial population as a representative of the total bioaerosols (viruses, bacteria, and fungi) has been raised frequently because of the high reponse speed, resolution, and compatibility with culture-based bioaerosol monitoring. On the other hand, additional engineering attempts are required to confer significance because of the size-classified (bioluminescence for different PM sizes) and specific (bioluminescence per unit PM mass) biological risks of air for providing proper interventions in the case of airborne transmission. In this study, disc-type impactors to cut-off aerosols larger than 1 µm, 2.5 µm, and 10 µm were designed and constructed to collect PM1, PM2.5, and PM10 on sampling swabs. This engineering enabled reliable size-classified bioluminescence signals using a commercial ATP luminometer after just 5 min of air intake. The simultaneous operations of a six-stage Andersen impactor and optical PM spectrometers were conducted to determine the correlations between the resulting RLU and colony forming unit (CFU; from the Andersen impactor) or PM mass concentration (deriving specific bioluminescence).


Assuntos
Técnicas Biossensoriais , COVID-19 , Humanos , Trifosfato de Adenosina/análise , Pandemias , Microbiologia do Ar , Técnicas Biossensoriais/métodos , COVID-19/diagnóstico , Aerossóis e Gotículas Respiratórios , Bactérias , Fungos , Monitoramento Ambiental/métodos , Tamanho da Partícula
13.
Anal Biochem ; 663: 115021, 2023 02 15.
Artigo em Inglês | MEDLINE | ID: mdl-36539047

RESUMO

ATP is a high-energy compound that plays a vital role in biological metabolism. Abnormal changes in ATP concentration are related to various diseases and reflect microbial metabolism in biofilms. In this work, we prepared carbon quantum dots (CDs) with aggregation-induced fluorescence inhibition effect using the bacterial culture medium as raw material with a hydrothermal method. Then, an abiotic fluorescent nanoprobe named CDs@zeolitic imidazolate frameworks-90 (ZIF-90) was facilely synthesized by encapsulating CDs into ZIF-90. Owing to the encapsulation of CDs in the hollow structure of ZIF-90, the blue fluorescence emission of CDs@ZIF-90 decreased significantly. In the presence of ATP, the ZIF-90 framework was destroyed due to the strong coordination between ATP and Zn2+. The released CDs exhibited stronger fluorescence intensity, which was closely related to the ATP concentration. The convenient synthesis process and rapid ATP-responsive ability make CDs@ZIF-90 highly promising for clinical and environmental analysis.


Assuntos
Pontos Quânticos , Zeolitas , Trifosfato de Adenosina/análise , Corantes Fluorescentes/química , Carbono/química , Pontos Quânticos/química
14.
Braz. J. Pharm. Sci. (Online) ; 59: e21220, 2023. graf
Artigo em Inglês | LILACS | ID: biblio-1439533

RESUMO

Abstract Exposure to methanol can cause serious consequences such as permanent visual disturbances and death. The heart tissue is highly vulnerable to ATP deficiency. Our study aimed to investigate whether exogenous ATP administration may alleviate methanol-induced ATP deficiency and subsequent oxidative damage in rat heart tissue. A total of 30 rats were divided into equal five groups; Healthy Group (HG), Methotrexate (MXG), Methanol (MeOH), Methotrexate+Methanol (MXM), and Methotrexate+Methanol+ATP (MMA) groups. We inhibited tetrahydrofolate synthesis by methotrexate to induce methanol toxicity. Methotrexate was administered to MXG, MXM, and MMA group animals for seven days with a catheter directly to the stomach at a 0,3 mg/kg dose per day. At the end of this period, % 20 methanol at a dose of 3 g/kg was administered to MeOH, MMA and MXM group animals. Immediately after methanol application, MMA group animals were injected with ATP at a 4 mg/kg dose intraperitoneally. Blood samples and heart tissues were used for biochemical analysis and histopathological examination. Co-exposure to methanol and methotrexate substantially exacerbated cardiac damage, indicating the potent cardiotoxic effects of methanol. However, the administration of exogenous ATP to MMA group animals brought biochemical oxidative damage parameters and histopathological findings closer to HG.


Assuntos
Animais , Masculino , Ratos , Trifosfato de Adenosina/análise , Metanol/efeitos adversos , Cardiotoxicidade/classificação
15.
Anal Chem ; 94(42): 14699-14706, 2022 10 25.
Artigo em Inglês | MEDLINE | ID: mdl-36245090

RESUMO

In situ and quantitative measurements of adenosine 5'-triphosphate (ATP) in single living cells are highly desired for understanding several sorts of necessary physiological and pathological processes. Due to its small size and high sensitivity, an ultra-microelectrode can be used for single-cell analysis. However, ATP is difficult to detect in single cells because it is nonelectroactive and low in content. Herein, we introduced an electrochemical nano-biosensor based on an amphiphilic aptamer-assisted carbon fiber nanoelectrode (aptCFNE) with high signal-to-noise ratio. The low current (e.g., 60 pA) and the tiny diameter of the tip (ca. 400 nm) of the nanosensor made it noninvasive to living cells. The amphiphilic aptamer has good biocompatibility and can be stably modified to the surface of functionalized electrodes. CFNE, which was modified with ferrocene-labeled aptamer, could quickly and selectively detect ATP content in the nucleus, cytoplasm, and extracellular space of single HeLa cells. The results showed that the ATP contents in the nucleus, cytoplasm, and extracellular space were 568 ± 9, 461 ± 20, and 312 ± 4 µM, respectively. The anticancer drug treatment effects on the cellular level were further recorded, which was of great significance for understanding ATP-related biological processes and drug screenings. This strategy is universally applicable to detect other targets by changing the aptamer sequence, which will greatly improve our understanding of cell heterogeneity and provide a more reliable scientific basis for exploring major diseases at the single-cell level.


Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Humanos , Aptâmeros de Nucleotídeos/química , Metalocenos , Fibra de Carbono , Células HeLa , Técnicas Biossensoriais/métodos , Trifosfato de Adenosina/análise , Adenosina
16.
Sci Rep ; 12(1): 17946, 2022 10 26.
Artigo em Inglês | MEDLINE | ID: mdl-36289258

RESUMO

In fluctuating environments, many microorganisms acquire phenotypic heterogeneity as a survival tactic to increase the likelihood of survival of the overall population. One example of this interindividual heterogeneity is the diversity of ATP concentration among members of Escherichia coli populations under glucose deprivation. Despite the importance of such environmentally driven phenotypic heterogeneity, how the differences in intracellular ATP concentration emerge among individual E. coli organisms is unknown. In this study, we focused on the mechanism through which individual E. coli achieve high intracellular ATP concentrations. First, we measured the ATP retained by E. coli over time when cultured at low (0.1 mM) and control (22.2 mM) concentrations of glucose and obtained the chronological change in ATP concentrations. Then, by comparing these chronological change of ATP concentrations and analyzing whether stochastic state transitions, periodic oscillations, cellular age, and intercellular communication-which have been reported as molecular biological mechanisms for generating interindividual heterogeneity-are involved, we showed that the appearance of high ATP-holding individuals observed among E. coli can be explained only by intercellular transmission. By performing metabolomic analysis of post-culture medium, we revealed a significant increase in the ATP, especially at low glucose, and that the number of E. coli that retain significantly higher ATP can be controlled by adding large amounts of ATP to the medium, even in populations cultured under control glucose concentrations. These results reveal for the first time that ATP-mediated intercellular transmission enables some individuals in E. coli populations grown at low glucose to retain large amounts of ATP.


Assuntos
Escherichia coli , Glucose , Humanos , Glucose/análise , Comunicação Celular , Trifosfato de Adenosina/análise
17.
Mikrochim Acta ; 189(11): 423, 2022 10 18.
Artigo em Inglês | MEDLINE | ID: mdl-36255523

RESUMO

A novel ratiometric electrochemiluminescence (ECL) system based on gold nanostars (AuNSs) support was constructed for the determination of hypotonicity-induced ATP release from HepG2 cells. AuNS@Lu nanoprobe was used as anodic luminophore and K2S2O8 as cathodic luminophore as well as anodic co-reactant. AuNS with the large specific surface was adopted to adsorb plentiful luminol to form solid-state probe and as affinity support to immobilize ATP aptamer (Apt). The obtained nanocomposite (Apt-AuNS@Lu) generated a strong ECL signal at + 0.4 V (vs. Ag/AgCl) with co-reactant K2S2O8, because of excellent conductivity and catalytic activity of AuNS. Furthermore, graphene oxide was reduced onto indium tin oxide (ITO) electrodes to facilitate the electron transfer. Following, polydopamine (PDA) film was formed via self-polymerization, improving stability and adhesion of the electrode surface. To immobilize ATP capture aptamer (AptC), abounding AuNSs were attached to RGO/PDA surface. When the sensor was incubated in the mixture solution of Apt-AuNS@Lu and target ATP, the ECL signal of Apt-AuNS@Lu increased with the increase of ATP concentration, meanwhile, the signal of K2S2O8 declined. The ratio of the two luminophores was used for the quantitative determination of ATP. The linear range was 5 to 250 nM, and the limit of detection was 1.4 nM at (3σ)/S. The method was successfully applied to analyze ATP release from HepG2 cells stimulated by 0.45% NaCl hypotonic solution. The results showed that the release kinetics profile of ATP had a sigmoidal shape with rapid release within 10 min and then slowed. Compared to the isotonic groups, the intracellular ATP concentration was 3.7 ± 0.3 µM (n = 3) decreasing by 40.3% and the extracellular was 23.4 ± 1.2 nM (n = 3) increasing by 9.2 times in the hypotonicity for 10 min, which showed ATP release from cells and good agreement with commercial ELISA test. The proposed strategy would be beneficial to broadening application of ECL technology in studying cell biological functions.


Assuntos
Luminol , Nanopartículas Metálicas , Medições Luminescentes , Cloreto de Sódio , Soluções Hipotônicas , Ouro , Trifosfato de Adenosina/análise
18.
Front Endocrinol (Lausanne) ; 13: 999475, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36246901

RESUMO

Particulate matter (PM) in polluted air can be exposed to the human body through inhalation, ingestion, and skin contact, accumulating in various organs throughout the body. Organ accumulation of PM is a growing health concern, particularly in the cardiovascular system. PM emissions are formed in the air by solid particles, liquid droplets, and fuel - particularly diesel - combustion. PM2.5 (size < 2.5 µm particle) is a major risk factor for approximately 200,000 premature deaths annually caused by air pollution. This study assessed the deleterious effects of diesel-derived PM2.5 exposure in HL-1 mouse cardiomyocyte cell lines. The PM2.5-induced biological changes, including ultrastructure, intracellular reactive oxygen species (ROS) generation, viability, and intracellular ATP levels, were analyzed. Moreover, we analyzed changes in transcriptomics using RNA sequencing and metabolomics using gas chromatography-tandem mass spectrometry (GC-MS/MS) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) in PM2.5-treated HL-1 cells. Ultrastructural analysis using transmission electron microscopy revealed disruption of mitochondrial cristae structures in a PM2.5 dose-dependent manner. The elevation of ROS levels and reduction in cell viability and ATP levels were similarly observed in a PM2.5 dose-dependently. In addition, 6,005 genes were differentially expressed (fold change cut-off ± 4) from a total of 45,777 identified genes, and 20 amino acids (AAs) were differentially expressed (fold change cut-off ± 1.2) from a total of 28 identified AAs profiles. Using bioinformatic analysis with ingenuity pathway analysis (IPA) software, we found that the changes in the transcriptome and metabolome are highly related to changes in biological functions, including homeostasis of Ca2+, depolarization of mitochondria, the function of mitochondria, synthesis of ATP, and cardiomyopathy. Moreover, an integrated single omics network was constructed by combining the transcriptome and the metabolome. In silico prediction analysis with IPA predicted that upregulation of mitochondria depolarization, ROS generation, cardiomyopathy, suppression of Ca2+ homeostasis, mitochondrial function, and ATP synthesis occurred in PM2.5-treated HL-1 cells. In particular, the cardiac movement of HL-1 was significantly reduced after PM2.5 treatment. In conclusion, our results assessed the harmful effects of PM2.5 on mitochondrial function and analyzed the biological changes related to cardiac movement, which is potentially associated with cardiovascular diseases.


Assuntos
Poluentes Atmosféricos , Material Particulado , Trifosfato de Adenosina/análise , Trifosfato de Adenosina/metabolismo , Trifosfato de Adenosina/farmacologia , Poluentes Atmosféricos/análise , Poluentes Atmosféricos/toxicidade , Aminoácidos/metabolismo , Animais , Cromatografia Líquida , Humanos , Camundongos , Mitocôndrias/metabolismo , Miócitos Cardíacos/metabolismo , Material Particulado/análise , Material Particulado/toxicidade , Espécies Reativas de Oxigênio/metabolismo , Espectrometria de Massas em Tandem
19.
J Air Waste Manag Assoc ; 72(12): 1327-1340, 2022 12.
Artigo em Inglês | MEDLINE | ID: mdl-36226866

RESUMO

Bioaerosols play a momentous role in the transmission of human infectious diseases, so there has been increasing concern over their exposure in recent years. Bioaerosol monitor is crucial in environmental fields. Based on the universal existence of Adenosine triphosphate (ATP) in bioaerosols, ATP bioluminescence can be used as a powerful technique to detect bioaerosols without interference from non-bioaerosols. When ATP is released from bioaerosols, they can quantify microbial biomass by ATP bioluminescence. In this review, we provide the latest methodological improvements that enable more reliable quantification of bioaerosols in complex environmental samples, especially the use of ATP bioluminescence in this era of technological advancement via the following routes: lower sample content for the trace existence of bioaerosols in the atmosphere, higher sensitivity of ATP bioluminescence reaction system and shorter process times. We also highlight the new techniques in improving the efficiencies of these monitoring processes. The purpose of this paper is to make more people realize the great potential of the ATP bioluminescence system for monitoring airborne microorganisms. Additionally, the present work intends to increase people's awareness of developing novel technology combined with ATP bioluminescence reaction system to realize rapid, real-time, and sensitive sensing of bioaerosols.Implications: The ATP bioluminescence methodology can not only eliminate the interference of co-existing nonbiological (fluorescent or PM) but also significantly improve the efficiency of bioaerosol. Recent progresses, such as the application of ATP fluorescence technology in bioaerosol monitoring, indicating that the efficiency and sensitivity are possible to be further improved. Nevertheless, there is no reviews address these advances and deeply analyze the application of ATP fluorescence technology in this field. his contribution will attract wide attention from both academic and industrial communities of this field, as well as researchers engaging in environmental monitoring. Furthermore, the strategies and techniques of studying the ATP bioluminescence reviewed here is instructive for environment monitoring in various fields. Therefore, in view of significance and broad interest, we feel strongly that our critical review is very essential to the field of public health security, pharmaceutics, anti-bioterrorism, etc., and would like it to be published in Journal of the Air & Waste Management Association.


Assuntos
Trifosfato de Adenosina , Microbiologia do Ar , Humanos , Aerossóis/análise , Trifosfato de Adenosina/análise , Atmosfera , Monitoramento Ambiental/métodos
20.
ACS Appl Mater Interfaces ; 14(36): 40624-40632, 2022 Sep 14.
Artigo em Inglês | MEDLINE | ID: mdl-36049088

RESUMO

As an energy supplier, ATP plays an important role in various life activities, and there is an urgent need to develop an effective means of detecting ATP. However, the traditional sensors face serious nonspecific adsorption. In this work, an antifouling electrochemical biosensor based on the interpenetrating network of Y-DNA scaffold and polyaniline hydrogel was designed for ATP detection. The polyaniline hydrogel was conducive to the transport of electrons and ions, the structure of Y-DNA cross-linked by ATP aptamers in the polyaniline hydrogel achieved the effect of signal amplification. Super hydrophilic cellulose nanocrystals (CNCs) and zwitterion polypeptide sequence (Pep) were doped to play a synergistic antifouling effect. The hydrogel sensor we have built has a wide linear range of 0.1 pM-1 µM for ATP detection and a low detection limit of 0.025 pM (S/N = 3). For ATP detection in actual serum samples, the recovery of this sensor was 99.5%-106%, and the relative standard deviation was 0.4%-2.88%. It is proven that the sensor has good ATP detection performance, and it will provide a certain reference value for the detection of other biological small molecules.


Assuntos
Incrustação Biológica , Técnicas Biossensoriais , Trifosfato de Adenosina/análise , Compostos de Anilina , Incrustação Biológica/prevenção & controle , DNA/química , Técnicas Eletroquímicas , Hidrogéis , Limite de Detecção
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