RESUMO
Cobalt-doped Prussian blue composite nanocubes (Co-PB NCs) were synthesized, which can quickly convert O2 to O2â¢- and 1O2. Due to the presence of cobalt and iron transition metal redox electron pairs, Co-PB NCs with high oxidase mimetic activity can rapidly oxidize the substrate 3,3',5,5'-tetramethylbenzidine (TMB) to produce blue products (ox-TMB) without the assistance of unstable H2O2. Using ascorbic acid-2-phosphate trisodium salt (AAP) as a substrate, it can be converted to reduced ascorbic acid (AA) under acid phosphatase (ACP) hydrolysis, resulting in suppression of TMB oxidation. Therefore, an enzyme cascade signal amplification strategy for rapid colorimetric detection of AA/ACP was developed based on the high-efficiency oxidase-like activity of Co-PB NCs combined with the hydrolysis effect of ACP. The color changes at low concentrations of AA and ACP could be observed by the naked eye, and the detection limits of AA and ACP were 1.67 µM and 0.0266 U/L, respectively. The developed colorimetric method was applied to the determination of AA in beverages and ACP in human serum, and the RSDs were less than 3%, showing good reproducibility. This work provides a promising strategy for the use of metal-doped Prussian blue composite material for the construction of rapid colorimetric sensing platforms that avoid the use of unstable hydrogen peroxide.
Assuntos
Fosfatase Ácida , Ácido Ascórbico , Cobalto , Colorimetria , Ferrocianetos , Limite de Detecção , Colorimetria/métodos , Ácido Ascórbico/química , Ácido Ascórbico/sangue , Ácido Ascórbico/análise , Ácido Ascórbico/análogos & derivados , Ferrocianetos/química , Humanos , Fosfatase Ácida/sangue , Fosfatase Ácida/análise , Fosfatase Ácida/química , Cobalto/química , Benzidinas/química , Peróxido de Hidrogênio/química , Oxirredução , Sucos de Frutas e Vegetais/análiseRESUMO
Highly stable copper nanocluster (CuNCs) with aggregation-induced emission (AIE) properties was synthesized. α-, ß-, and γ- MnO2 were utilized as quenchers, with CuNCs fluorescence quenching of 48.9%, 91.5%, and 96.6%, respectively. L-ascorbate-2-phosphate (AAP) was hydrolyzed by acid phosphatase (ACP), and ascorbic acid (AA) was formed. Then, MnO2 could be restored by AA, and the fluorescence of the CuNCs could be restored. An on-off-on detection platform with a high signal/noise ratio was constructed for the sensing of ACP. The fluorescence recovery rate of the CuNCs was related to the crystal forms of MnO2. Then, the equilibrium constants (K) for the reaction between AA and MnO2 were calculated to evaluate the reaction process. Compared with the K values of CuNCs/α-MnO2 and CuNCs/γ-MnO2, the K values for AA and ß-MnO2 were maximum. The CuNCs/ß-MnO2 system exhibited optimal fluorescence recovery for the sensitive detection of ACP. In the concentration range 0.005-0.06 U/mL, the detection limit was 0.0028 U/mL. The determination of serum ACP levels also revealed satisfactory results. This study provides novel insights into enhancing the sensitivity of the determination of quenchers in different crystal form.
Assuntos
Fosfatase Ácida , Cobre , Compostos de Manganês , Nanopartículas Metálicas , Óxidos , Compostos de Manganês/química , Óxidos/química , Cobre/química , Fosfatase Ácida/química , Fosfatase Ácida/metabolismo , Nanopartículas Metálicas/química , Ácido Ascórbico/química , Ácido Ascórbico/análogos & derivados , Limite de Detecção , Espectrometria de Fluorescência/métodos , HumanosRESUMO
Ectophosphatases catalyse the hydrolysis of phosphorylated molecules, such as phospho-amino acids, in the extracellular environment. Nevertheless, the hydrolysis of nucleotides in the extracellular environment is typically catalysed by ectonucleotidases. Studies have shown that acid ectophosphatase, or transmembrane-prostatic acid phosphatase (TM-PAP), a membrane-bound splice variant of prostatic acid phosphatase, has ecto-5'-nucleotidase activity. Furthermore, it was demonstrated that ectophosphatase cannot hydrolyse ATP, ADP, or AMP in triple-negative breast cancer cells. In contrast to previous findings in MDA-MB-231 cells, the ectophosphatase studied in the present work displayed a remarkable capacity to hydrolyse AMP in luminal A breast cancer cells (MCF-7). We showed that AMP dose-dependently inhibited p-nitrophenylphosphate (p-NPP) hydrolysis. The p-NPP and AMP hydrolysis showed similar biochemical behaviours, such as increased hydrolysis under acidic conditions and comparable inhibition by NiCl2, ammonium molybdate, and sodium orthovanadate. In addition, this ectophosphatase with ectonucleotidase activity was essential for the release of adenosine and inorganic phosphate from phosphorylated molecules available in the extracellular microenvironment. This is the first study to show that prostatic acid phosphatase on the membrane surface of breast cancer cells (MCF-7) is correlated with cell adhesion and migration.
Assuntos
Fosfatase Ácida , Neoplasias da Mama , Humanos , Células MCF-7 , Feminino , Hidrólise , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Neoplasias da Mama/enzimologia , Fosfatase Ácida/metabolismo , 5'-Nucleotidase/metabolismo , Monofosfato de Adenosina/metabolismo , Proteínas Tirosina Fosfatases/metabolismo , Proteínas Tirosina Fosfatases/antagonistas & inibidores , Nitrofenóis/farmacologia , Nitrofenóis/metabolismo , Linhagem Celular Tumoral , Compostos OrganofosforadosRESUMO
PURPOSE: The acid phosphatase 1 (ACP1) gene encodes low-molecular-weight protein tyrosine phosphatase, which is overexpressed in prostate cancer (PC) and a potential therapeutic target. We analyzed ACP1 expression in primary/metastatic PC and its association with molecular profiles and clinical outcomes. METHODS: NextGen sequencing of DNA (592-gene/whole-exome sequencing)/RNA(whole-transcriptome sequencing) was performed for 5,028 specimens. ACP1-High/ACP1-Low expression was defined as quartile (Q4/1) of RNA transcripts per million (TPM). DNA mutational profiles were analyzed for ACP1-quartile-stratified samples. Gene set enrichment analysis was used for Hallmark collection of pathways. PD-L1+(≥2+, ≥5%; SP142) was tested by immunohistochemistry. Tumor microenvironment's (TME) immune cell fractions were estimated by RNA deconvolution/quanTIseq. Overall survival (OS) was assessed from initial diagnosis/treatment initiation to death/last follow-up. RESULTS: We included 3,058 (60.8%) samples from the prostate, 634 (12.6%) from lymph node metastases (LNMs), and 1,307 (26.0%) from distant metastases (DMs). ACP1 expression was higher in LNM/DM than prostate (49.8/47.9 v 44.1 TPM; P < .0001). TP53 mutations were enriched in ACP1-Q4 (37.9%[Q4] v 27.0%[Q1]; P < .001) among prostate samples. Pathways associated with cell cycle regulation and oxidative phosphorylation were enriched in ACP1-Q4, whereas epithelial-mesenchymal transition and tumor necrosis factor-alpha signaling via nuclear factor kappa-light-chain-enhancer of activated B-cell pathways were enriched in ACP1-Q1. Neuroendocrine and androgen receptor signaling was increased in ACP1-Q4. M2 macrophages and natural killer cell fractions were increased, whereas T cells and M1 macrophages were decreased in ACP1-Q4. While OS differences between ACP1-Q1/Q4 were not statistically significant, there was a trend for worse OS among ACP1-Q4 prostate samples (Q4 v Q1: hazard ratio [HR], 1.19 [95% CI, 0.99 to 1.42]; P = .06) and DM (HR, 1.12 [95% CI, 0.93 to 1.36]; P = .22) but not LNM (HR, 0.98 [95% CI, 0.74 to 1.29]; P = .87). CONCLUSION: ACP1-High tumors exhibit a distinct molecular profile and cold TME, highlighting ACP1's potential role in PC pathogenesis and novel therapeutic targeting.
Assuntos
Fosfatase Ácida , Neoplasias da Próstata , Humanos , Masculino , Neoplasias da Próstata/genética , Neoplasias da Próstata/patologia , Fosfatase Ácida/genética , Pessoa de Meia-Idade , Idoso , Proteínas Proto-Oncogênicas/genética , Proteínas Tirosina Fosfatases/genética , MutaçãoRESUMO
We have previously reported two single-agent phase I trials, evaluating the dose or schedule, of a DNA vaccine (pTVG-HP) encoding prostatic acid phosphatase (PAP) administered with GM-CSF as the adjuvant. These were in patients with PSA-recurrent, radiographically non-metastatic, prostate cancer (PCa). We report here the long-term safety and overall survival of these patients. Specifically, 22 patients with non-metastatic, castration-sensitive PCa (nmCSPC) were treated with pTVG-HP, 100-1500 µg, administered over 12 weeks and followed for 15 y. 17 patients with non-metastatic castration-resistant PCa (nmCRPC) were treated with 100 µg pTVG-HP with different schedules of administration over 1 y and followed for 5 y. No adverse events were detected in long-term follow-up from either trial that were deemed possibly related to vaccination. Patients with nmCSPC had a median overall survival of 12.3 y, with 5/22 (23%) alive at 15 y. 8/22 (36%) died due to prostate cancer with a median survival of 11.0 y, and 9/22 (41%) died of other causes. Patients with nmCRPC had a median overall survival of 4.5 y, with 8/17 (47%) alive at 5 y. The presence of T-cells specific for the PAP target antigen was detectable in 6/10 (60%) individuals with nmCSPC, and 3/5 (60%) individuals with nmCRPC, many years after immunization. The detection of immune responses to the vaccine target years after immunization suggests durable immunity can be elicited in patients using a DNA vaccine encoding a tumor-associated antigen.Trial Registration: NCT00582140 and NCT00849121.
Assuntos
Vacinas Anticâncer , Antígeno Prostático Específico , Neoplasias da Próstata , Vacinas de DNA , Humanos , Masculino , Vacinas de DNA/imunologia , Vacinas de DNA/administração & dosagem , Vacinas de DNA/efeitos adversos , Antígeno Prostático Específico/imunologia , Vacinas Anticâncer/imunologia , Vacinas Anticâncer/efeitos adversos , Vacinas Anticâncer/administração & dosagem , Idoso , Seguimentos , Neoplasias da Próstata/imunologia , Pessoa de Meia-Idade , Adjuvantes Imunológicos/administração & dosagem , Adjuvantes Imunológicos/efeitos adversos , Resultado do Tratamento , Idoso de 80 Anos ou mais , Fator Estimulador de Colônias de Granulócitos e Macrófagos/imunologia , Recidiva Local de Neoplasia , Análise de Sobrevida , Fosfatase Ácida , Proteínas Tirosina Fosfatases/imunologiaRESUMO
Engineering enzyme-substrate binding pockets is the most efficient approach for modifying catalytic activity, but is limited if the substrate binding sites are indistinct. Here, we developed a 3D convolutional neural network for predicting protein-ligand binding sites. The network was integrated by DenseNet, UNet, and self-attention for extracting features and recovering sample size. We attempted to enlarge the dataset by data augmentation, and the model achieved success rates of 48.4%, 35.5%, and 43.6% at a precision of ≥50% and 52%, 47.6%, and 58.1%. The distance of predicted and real center is ≤4 Å, which is based on SC6K, COACH420, and BU48 validation datasets. The substrate binding sites of Klebsiella variicola acid phosphatase (KvAP) and Bacillus anthracis proline 4-hydroxylase (BaP4H) were predicted using DUnet, showing high competitive performance of 53.8% and 56% of the predicted binding sites that critically affected the catalysis of KvAP and BaP4H. Virtual saturation mutagenesis was applied based on the predicted binding sites of KvAP, and the top-ranked 10 single mutations contributed to stronger enzyme-substrate binding varied while the predicted sites were different. The advantage of DUnet for predicting key residues responsible for enzyme activity further promoted the success rate of virtual mutagenesis. This study highlighted the significance of correctly predicting key binding sites for enzyme engineering.
Assuntos
Aprendizado de Máquina , Sítios de Ligação , Engenharia de Proteínas/métodos , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Fosfatase Ácida/química , Fosfatase Ácida/genética , Fosfatase Ácida/metabolismo , Especificidade por Substrato , Bacillus anthracis/genética , Bacillus anthracis/enzimologia , Klebsiella/genética , Klebsiella/enzimologia , Ligantes , Ligação Proteica , Modelos Moleculares , Redes Neurais de ComputaçãoRESUMO
We performed a comprehensive study of protein (total protein, medium-molecular-weight peptides, creatinine, and urea), purine (uric acid), and lipid (cholesterol, triglycerides) metabolism, activity of AST, ALT, and acid phosphatase in blood plasma of white male rats under conditions of restriction of motor activity up to 28 days. Patterns of changes in metabolic profile during hypokinesia were established: prevalence of catabolic processes and atherogenic shifts in the lipid spectrum with maximum manifestation on 14-21 days of the experiment.
Assuntos
Colesterol , Triglicerídeos , Animais , Masculino , Ratos , Triglicerídeos/sangue , Triglicerídeos/metabolismo , Colesterol/sangue , Colesterol/metabolismo , Ácido Úrico/sangue , Ácido Úrico/metabolismo , Atividade Motora/fisiologia , Metaboloma/fisiologia , Metabolismo dos Lipídeos/fisiologia , Aspartato Aminotransferases/sangue , Aspartato Aminotransferases/metabolismo , Alanina Transaminase/sangue , Alanina Transaminase/metabolismo , Creatinina/sangue , Fosfatase Ácida/metabolismo , Fosfatase Ácida/sangue , Ureia/sangue , Hipocinesia/metabolismo , Hipocinesia/fisiopatologiaRESUMO
Microorganisms produce extracellular enzymes to meet elemental requirements and cope with stoichiometric imbalances of resources. To gain insights into the cycling of C, N, and P, the activities of the Câ¶Nâ¶P acquisition enzymes have been extensively investigated. To detect the effects of long-term fertilization practices on soil nutrient balance and characteristics of soil enzymatic stoichiometry in black soil, four different fertilization treatments were selectedï¼ no fertilization ï¼CKï¼, nitrogen fertilizer ï¼Nï¼, phosphorus fertilizer ï¼Pï¼, and combination of nitrogen and phosphorus fertilizers ï¼NPï¼. Soil samples were collected in both April 2021 and April 2022 to determine soil enzyme activities and their stoichiometric characteristics. The results showed that soil acid phosphatase and ß-D-glucosidase activities were significantly higher in the N and NP treatments than in CK by 68%-158% and 26%-222%, respectively. Soil ß-N-acetylaminoglucosidase activities were significantly higher in the P and NP treatments, with the highest around 75.48 nmol·ï¼g·hï¼-1 and 106.81 nmol·ï¼g·hï¼-1, respectively. Two-way ANOVA analysis showed that N and P inputs had a great impact on soil enzyme activities. Redundancy analysis showed that the main factors controlling enzyme activities were soil pH, microbial biomass phosphorus, and soil available P content. It was found that N inputs significantly increased enzyme vector length, which was ranged from 1.32 to 1.52, and the enzyme vector angles were all larger than 45°, suggesting C and P co-limited in the black soils. These findings suggest that 40 years of fertilization have had a great impact on soil enzymes and the related resource use strategy, which provides great implications for assessing soil nutrients balance and soil sustainability.
Assuntos
Fertilizantes , Nitrogênio , Fósforo , Microbiologia do Solo , Solo , Solo/química , Fósforo/análise , Fosfatase Ácida/metabolismo , Carbono/análise , Fatores de Tempo , ChinaRESUMO
The development of high-accuracy technologies to distinguish the quite tiny concentration change of tumor markers between negative and positive is of vital significance for early screening and diagnosis of cancers, but is still a great challenge for the conventional biosensors because of their "gradual" detection mode. Herein, a unique "leap-type" responsive lanthanide MOF-based biosensor (designated as Tb-CeMOF-X) with defect-mediated redox-/photo-activities is developed for precisely identifying acid phosphatase (ACP), an early pathological marker of prostate cancer (PCa) in serum. The engineered Tb-CeMOF-X probe achieves a bursting switch-on luminescence at the critical concentration of ACP (9â U â L-1), while keeping silent below this threshold, undergoing a qualitative signal change from "zero" to "one" between negative and positive indicators and thus significantly improving the identification precision. Significantly, such "leap-type" response performance can be further edited and amplified by rational defect engineering in the crystal structure to improve the accessibility of active centers, consequently maximizing the detection sensitivity toward ACP in the complex biological media. This study proposes the first paradigm for the development of "leap-type" biosensors with ultra-sensitive differentiation capability between negative and positive, and provides a potentially valuable tool for early and accurate screening of PCa.
Assuntos
Elementos da Série dos Lantanídeos , Estruturas Metalorgânicas , Oxirredução , Neoplasias da Próstata , Estruturas Metalorgânicas/química , Neoplasias da Próstata/diagnóstico , Neoplasias da Próstata/metabolismo , Masculino , Humanos , Elementos da Série dos Lantanídeos/química , Técnicas Biossensoriais/métodos , Fosfatase Ácida/química , Fosfatase Ácida/metabolismo , Fosfatase Ácida/sangue , Detecção Precoce de Câncer , Processos FotoquímicosRESUMO
The purple acid phosphatase was purified from 5.9-fold to apparent homogeneity from Anagelis arvensis seeds using SP-Sephadex C-50 and Sephadex G-100 chromatography. The results of residual activity tests conducted using different temperature ranges (50-70 °C) were calculated as the activation energy (Ed = 72 kJ/mol), enthalpy (69.31 ≤ (ΔH° ≤ 69.10 kJ/mol), entropy (-122.48 ≤ ΔS° ≤ -121.13 J/mol·K), and Gibbs free energy (108.87 ≤ ΔG° ≤ 111.25 kJ/mol) of the enzyme irreversible denaturation. These thermodynamic parameters indicate that this novel PAP is highly thermostable and may be significant for use in industrial applications. However, it may be confirmed by stopped-flow measurements that this substitution produces a chromophoric Fe3+ site and a Pi-substrate interaction that is about ten times faster. Additionally, these data show that phenyl phosphate hydrolysis proceeds more rapidly in metal form of A. arvensis PAP than the creation of a µ-1,3 phosphate complex. The Fe3+ site in the native Fe3+-Mn2+ derivative interacts with it at a faster rate than in the Fe3+-Fe2+ form. This is most likely caused by a network of hydrogen bonds between the first and second coordination spheres. This suggests that the choice of metal ions plays a significant role in regulating the activity of this enzyme.
Assuntos
Fosfatase Ácida , Domínio Catalítico , Termodinâmica , Fosfatase Ácida/química , Fosfatase Ácida/metabolismo , Cinética , Especificidade por Substrato , Cátions Bivalentes , Ligação Proteica , Hidrólise , Concentração de Íons de Hidrogênio , Temperatura , Metais/químicaRESUMO
As part of the multifaceted strategies developed to shape the common environmental policy, considerable attention is now being paid to assessing the degree of environmental degradation in soil under xenobiotic pressure. Bisphenol A (BPA) has only been marginally investigated in this ecosystem context. Therefore, research was carried out to determine the biochemical properties of soils contaminated with BPA at two levels of contamination: 500 mg and 1000 mg BPA kg-1 d.m. of soil. Reliable biochemical indicators of soil changes, whose activity was determined in the pot experiment conducted, were used: dehydrogenases, catalase, urease, acid phosphatase, alkaline phosphatase, arylsulfatase, and ß-glucosidase. Using the definition of soil health as the ability to promote plant growth, the influence of BPA on the growth and development of Zea mays, a plant used for energy production, was also tested. As well as the biomass of aerial parts and roots, the leaf greenness index (SPAD) of Zea mays was also assessed. A key aspect of the research was to identify those of the six remediating substances-molecular sieve, zeolite, sepiolite, starch, grass compost, and fermented bark-whose use could become common practice in both environmental protection and agriculture. Exposure to BPA revealed the highest sensitivity of dehydrogenases, urease, and acid phosphatase and the lowest sensitivity of alkaline phosphatase and catalase to this phenolic compound. The enzyme response generated a reduction in the biochemical fertility index (BA21) of 64% (500 mg BPA) and 70% (1000 mg BPA kg-1 d.m. of soil). The toxicity of BPA led to a drastic reduction in root biomass and consequently in the aerial parts of Zea mays. Compost and molecular sieve proved to be the most effective in mitigating the negative effect of the xenobiotic on the parameters discussed. The results obtained are the first research step in the search for further substances with bioremediation potential against both soil and plants under BPA pressure.
Assuntos
Fosfatase Ácida , Compostos Benzidrílicos , Fenóis , Poluentes do Solo , Solo , Zea mays , Fenóis/química , Compostos Benzidrílicos/química , Poluentes do Solo/química , Zea mays/química , Solo/química , Fosfatase Ácida/metabolismo , Arilsulfatases/metabolismo , Fosfatase Alcalina/metabolismo , Zeolitas/química , Oxirredutases/metabolismo , Urease/metabolismo , Catalase/metabolismo , Biodegradação Ambiental , Silicatos de Magnésio/química , Amido/química , beta-Glucosidase/metabolismo , Compostagem/métodosRESUMO
The aim of this study was to modify phytase YiAPPA via protein surficial residue mutation to obtain phytase mutants with improved thermostability and activity, enhancing its application potential in the food industry. First, homology modeling of YiAPPA was performed. By adopting the strategy of protein surficial residue mutation, the lysine (Lys) and glycine (Gly) residues on the protein surface were selected for site-directed mutagenesis to construct single-site mutants. Thermostability screening was performed to obtain mutants (K189R and K216R) with significantly elevated thermostability. The combined mutant K189R/K216R was constructed via beneficial mutation site stacking and characterized. Compared with those of YiAPPA, the half-life of K189R/K216R at 80°C was extended from 14.81 min to 23.35 min, half-inactivation temperature (T50 30) was increased from 55.12°C to 62.44°C, and Tm value was increased from 48.36°C to 53.18°C. Meanwhile, the specific activity of K189R/K216R at 37°C and pH 4.5 increased from 3960.81 to 4469.13 U/mg. Molecular structure modeling analysis and molecular dynamics simulation showed that new hydrogen bonds were introduced into K189R/K216R, improving the stability of certain structural units of the phytase and its thermostability. The enhanced activity was primarily attributed to reduced enzyme-substrate binding energy and shorter nucleophilic attack distance between the catalytic residue His28 and the phytate substrate. Additionally, the K189R/K216R mutant increased the hydrolysis efficiency of phytate in food ingredients by 1.73-2.36 times. This study established an effective method for the molecular modification of phytase thermostability and activity, providing the food industry with an efficient phytase for hydrolyzing phytate in food ingredients.
Assuntos
6-Fitase , Estabilidade Enzimática , Mutagênese Sítio-Dirigida , 6-Fitase/genética , 6-Fitase/metabolismo , 6-Fitase/química , Simulação de Dinâmica Molecular , Engenharia de Proteínas , Concentração de Íons de Hidrogênio , Cinética , Ácido Fítico/metabolismo , Modelos Moleculares , Temperatura , Temperatura Alta , Mutação , Escherichia coli/genética , Escherichia coli/metabolismo , Indústria Alimentícia , Fosfatase Ácida , Proteínas de Escherichia coliRESUMO
Some research has identified the prevalence and motivation of using condoms by assailants during sexual assault cases proving the necessity of analyzing condom trace evidence. The majority of the papers published have discussed forensic analysis of lubricants from condoms retrieved at sexual assault scenes but those discussing the identification of semen from condoms are rare. Therefore, the present study aims to provide insight into the detectability of the semen that remained in a condom, to examine the effect of exposure time, environmental conditions, and condom type, and ultimately to determine the capability of the AP test and Microscopic examination for identification of this sample type. In the study, samples were collected from three male donors after being instructed on the proper way of collecting the semen sample. The received samples from the donors were checked first by microscopic examination to observe the sperm to confirm that the sample being handled was semen. After confirmation, samples were transferred to 4 prepared condoms (brands: dkt xxx and Manforce) and kept in conditions i.e. two condoms in a refrigerator maintained from 2 to 10°C and other ones at ambient temperature (weather status: summer season of average 39°C). The samples were analyzed into two batches, the first analysis batch was conducted after the samples were exposed to the conditions within 11-60 days. After analysis from the first batch, the samples were continuously kept in the same condition for the consecutive second batch conducted when the samples reached 40-90 days. This study has determined that semen remaining in a condom can be detected and each test studied is appropriate according to the exposure stage, i.e., time and conditions of exposure. It has been found that nonmotile spermatozoa can be observed when semen remains in the condom for a few days. However, if the sample reaches approximately 25 days at room temperature above 25°C or 54 days below 10°C, the semen may dry out limiting the effectiveness of microscopic examination. Despite this, even semen that remained in a condom for up to 90 days can be identified by Acid Phosphatase. Results on condom type used reveal that condom constituents can crossreact with semen but none of them can limit the semen identification with Acid Phosphatase.
Assuntos
Preservativos , Sêmen , Humanos , Masculino , Sêmen/química , Temperatura , Manejo de Espécimes , Fosfatase Ácida/análise , Microscopia , Fatores de TempoRESUMO
This study aimed to identify if biological material could be detected on the opposite side to deposition on fabric by commonly used presumptive and/or secondary tests. Additionally, this study aimed to ascertain if there is a difference in the DNA quantity and quality from samples obtained from both sides of the same substrate: cotton, polyester, denim, or combined viscose and polyester swatches. Blood, semen, or saliva (25 µL) was deposited on one side of 5 replicates of each fabric type and left for 24â¯h. Blood swatches were tested using Hemastix® and the ABACard® HemaTrace® immunoassay, semen swatches were tested using acid phosphatase (AP) reagent, the ABACard® p30® immunoassay and hematoxylin and eosin staining, and saliva swatches were tested using Phadebas® paper and the RSID-Saliva™ immunoassay. Both sides of each swatch were separately wet/dry swabbed and subjected to DNA analysis. Blood was able to be detected on the underside of all fabrics using both tests. Semen was able to be detected on the underside of swatches using the presumptive AP test but not p30®, and sperm was rarely observed. Saliva was able to be detected by RSID-Saliva™ but not Phadebas® paper when the underside of swatches were tested. Across all biological materials, DNA was able to be recovered from the top side of all 60 swatches. For the underside, DNA was able to be recovered from 54 swatches. Of the 6 swatches that DNA was unable to be recovered from, one sample was from semen and the rest were from saliva. This study has demonstrated that DNA and components of interest in forensically relevant biological material can be recovered from the opposite side to where it was originally deposited, and that observing biological material and/or DNA on one side of fabric does not definitively indicate direct deposition on that side.
Assuntos
Impressões Digitais de DNA , DNA , Saliva , Sêmen , Têxteis , Saliva/química , Sêmen/química , Humanos , Masculino , Projetos Piloto , DNA/análise , Imunoensaio , Sangue , Manchas de Sangue , Fosfatase Ácida/análise , VestuárioRESUMO
In this study, a nanozyme (ZIF-Co-Cys) with high oxidase-like catalytic activity was prepared, and a ratiometric fluorescent/photothermal dual-mode probe was constructed for organophosphorus pesticides (OPs) detection based on the competitive effect of ZIF-Co-Cys and the enzymatic reaction product of acid phosphatase (ACP) on o-phenylenediamine and the inhibition effect of OPs on ACP activity. Using dimethyl dichloroviny phosphate (DDVP) as the model, both the fluorescence intensity ratio and the temperature change of the probe solution exhibited an excellent correlation with OPs concentration. The detection limits were 1.64 ng/mL and 0.084 ng/mL, respectively. Additionally, the detection of DDVP residues in real samples verified the outstanding anti-interference and accuracy of the probe. This work not only provided a complementary dual-mode method for the accurate and rapid detection of OPs residues in complex samples, but also supplied a new insight into the design of a multi-mode sensing platform based on the cascade reaction of nanozyme.
Assuntos
Corantes Fluorescentes , Resíduos de Praguicidas , Corantes Fluorescentes/química , Resíduos de Praguicidas/análise , Resíduos de Praguicidas/química , Compostos Organofosforados/química , Compostos Organofosforados/análise , Fosfatase Ácida/análise , Fosfatase Ácida/química , Fosfatase Ácida/metabolismo , Espectrometria de Fluorescência/métodos , Limite de Detecção , Diclorvós/análise , TemperaturaRESUMO
Abiotic stress tends to induce oxidative damage to enzymes and organelles that in turns hampers the phosphorylation process and decreases the adenosine triphosphate (ATP) productivity. Artificial assemblies can alleviate abiotic stress and simultaneously provide nutrients to diminish the oxidative damage. Here, we have integrated natural acid phosphatase (ACP) and ATP synthase with plasmonic Au clusters in a biomimetic microreactor. ACP immobilized on the Au clusters is harnessed to generate proton influx to drive ATP synthase and concurrently supply phosphate to improve phosphorus availability to combat phosphorus-deficiency stress. In tandem with the reactive oxygen species (ROS) scavenging and the photothermal functionality of Au clusters, such an assembled microreactor exhibits an improved abiotic stress tolerance and achieves plasmon-accelerated ATP synthesis. This innovative approach offers an effective route to enhance the stress resistance of ATP synthase-based energy-generating systems, opening an exciting potential of these systems for biomimicking applications.
Assuntos
Mitocôndrias , Mitocôndrias/metabolismo , Ouro/química , Trifosfato de Adenosina/metabolismo , Trifosfato de Adenosina/química , Fosfatase Ácida/metabolismo , Fosfatase Ácida/química , Espécies Reativas de Oxigênio/metabolismo , ATPases Mitocondriais Próton-Translocadoras/metabolismoRESUMO
In this study, the effects of four types of amendments on effective Cd and Cd content in different parts of prickly ash soil and soil enzyme activity were studied, which provided scientific basis for acidification improvement of purple soil and heavy metal pollution control. A field experiment was conducted. Six treatments were set up:no fertilizer (CK), only chemical fertilizer (F), lime + chemical fertilizer (SF), organic fertilizer + chemical fertilizer (OM), biochar + chemical fertilizer (BF), and vinasse biomass ash + chemical fertilizer (JZ). Soil pH; available Cd (DTPA-Cd); Cd content in branches, leaves, shells, and seeds of Zanthoxylum; as well as the activities of catalase (S-CAT), acid phosphatase (S-ACP), and urease (S-UE) in different treatments were studied, and their relationships were clarified. The results showed following:â The two treatments of vinasse biomass ash + chemical fertilizer and lime + chemical fertilizer significantly increased soil pH (P < 0.05) to 3.39 and 2.25 units higher than that in the control, respectively. Compared with that in the control treatment, the content of available Cd in soil under vinasse biomass ash + chemical fertilizer and lime + chemical fertilizer treatment decreased by 28.91 % and 20.90 %, respectively. â¡ The contents of Cd in leaves, shells, and seeds of Zanthoxylum were decreased by 31.33 %, 30.24 %, and 34.01 %, respectively. The Cd enrichment ability of different parts of Zanthoxylum was different, with the specific performances being leaves > branches > seeds > shells. Compared with that of the control, the enrichment coefficient of each part of Zanthoxylum treated with vinasse biomass ash + chemical fertilizer decreased significantly(P < 0.05)by 27.54 %-40.0 %. ⢠The changes in catalase and urease activities in soil treated with amendments were similar. Compared with those in the control group, the above two enzyme activities were significantly increased by 191.26 % and 199.50 %, respectively, whereas the acid phosphatase activities were decreased by 16.45 %. Correlation analysis showed that soil available Cd content was significantly negatively correlated with soil pH value(P < 0.01), S-CAT and S-UE enzyme activities were significantly positively correlated with soil pH(P < 0.01), and the soil available Cd content was significantly negatively correlated (P < 0.01); the S-ACP enzyme showed the complete opposite trends. The application of lime and vinasse biomass ash to acidic purple soil had the most significant effect on neutralizing soil acidity. It was an effective measure to improve acidic purple soil and prevent heavy metal pollution by reducing the effective Cd content in soil and improving the soil environment while inhibiting the absorption and transfer of Cd in various parts of Zanthoxylum.
Assuntos
Cádmio , Fertilizantes , Poluentes do Solo , Solo , Poluentes do Solo/metabolismo , Cádmio/metabolismo , Solo/química , Urease/metabolismo , Zanthoxylum/química , Zanthoxylum/metabolismo , Fosfatase Ácida/metabolismo , Catalase/metabolismo , Disponibilidade Biológica , Óxidos/química , Compostos de Cálcio/química , Carvão Vegetal/químicaRESUMO
A colorimetric analysis platform has been successfully developed based on FeCo-NC dual-atom nanozyme (FeCo-NC DAzyme) for the detection of organophosphorus pesticides (OPPs). The FeCo-NC DAzyme exhibited exceptional oxidase-like activity (OXD), enabling the catalysis of colorless TMB to form blue oxidized TMB (oxTMB) without the need for H2O2 involvement. By combining acid phosphatase (ACP) hydrolase with FeCo-NC DAzyme, a "FeCo-NC DAzyme + TMB + ACP + SAP" colorimetric system was constructed, which facilitated the rapid detection of malathion. The chromogenic system was applied to detect malathion using a smartphone-based app and an auxiliary imaging interferogram device for colorimetric measurements, which have a linear range of 0.05-4.0 µM and a limit of detection (LOD) as low as 15 nM in real samples, comparable to UV-Vis and HPLC-DAD detection methods. Overall, these findings present a novel approach for convenient, rapid, and on-site monitoring of OPPs.
Assuntos
Colorimetria , Limite de Detecção , Praguicidas , Smartphone , Colorimetria/métodos , Praguicidas/análise , Compostos Organofosforados/análise , Compostos Organofosforados/química , Malation/análise , Malation/química , Oxirredutases/química , Ferro/química , Fosfatase Ácida/análise , Fosfatase Ácida/química , BenzidinasRESUMO
Dianthus barbatus linn. is widely used in gardens, mainly as flower beds and flower borders. The effects of different gradients of P on the growth and root morphology of Dianthus barbatus were studied to explore its morphological and physiological responses and adaptive strategies. Hence, this study provides a theoretical basis and practical guidance for D. barbatus production. Two soil substrates, namely loess and vegetable soil, and five phosphorus concentration gradients were set; no phosphorus application was used as the control. The morphology and physiology of D. barbatus were also investigated. Low-to-medium- and low-phosphorus treatments promoted the growth of D. barbatus in the above and underground parts of the plants grown on both substrates. Chlorophyll content, flower quantity, and acid phosphatase activity in the rhizosphere soil were significantly increased in the H1 and H2 treatments of loess and in the C4 treatment of vegetable soil. Thus, D. barbatus seems to reduce the damage caused by phosphorus stress by increasing chlorophyll content and root acid phosphatase activity. The latter was significantly higher in vegetable soil than in loess. Vegetable soil was more conducive to D. barbatus growth than loess.
Assuntos
Clorofila , Dianthus , Fósforo , Raízes de Plantas , Solo , Fósforo/metabolismo , Solo/química , Clorofila/metabolismo , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/metabolismo , Raízes de Plantas/efeitos dos fármacos , Dianthus/crescimento & desenvolvimento , Dianthus/metabolismo , Dianthus/fisiologia , Fosfatase Ácida/metabolismo , Flores/metabolismo , Flores/crescimento & desenvolvimento , RizosferaRESUMO
Developing water-soluble nanomaterials with high photoluminescence emission and high yield for biological analysis and imaging is urgently needed. Herein, water-soluble blue emitting silicon and nitrogen co-doped carbon dots (abbreviated as Si-CDs) of a high photoluminescence quantum yield of 80 % were effectively prepared with high yield rate (59.1 %) via one-step hydrothermal treatment of N-[3-(trimethoxysilyl)propyl]ethylenediamine (DAMO) and trans-aconitic acid. Furthermore, the Si-CDs demonstrate environmental robustness, photo-stability and biocompatibility. Given the importance of the potentially abnormal levels of acid phosphatase (ACP) in cancer diagnosis, developing a reliable and sensitive ACP measurement method is of significance for clinical research. The Si-CDs unexpectedly promote the catalytic oxidation of ACP on dopamine (DA) to polydopamine under acidic conditions through the produced reactive oxygen species (ROS). Correspondingly, a fluorescence response strategy using Si-CDs as the dual functions of probes and promoting enzyme activity of ACP on catalyzing DA was constructed to sensitively determine ACP. The quantitative analysis of ACP displayed a linear range of 0.1-60 U/L with a detection limit of 0.056 U/L. The accurate detection of ACP was successfully achieved in human serum through recovery tests. As a satisfactory fluorescent probe, Si-CDs were successfully applied to fluorescent imaging of A549 cells in cytoplasmic with long-term and safe staining. The Si-CDs have the dual properties of outstanding fluorescent probes and auxiliary oxidase activity, indicating their great potential in multifunctional applications.