ABSTRACT
Helicobacter pylori (H. pylori) is responsible for various chronic or acute diseases, such as stomach ulcers, dyspepsia, peptic ulcers, gastroesophageal reflux, gastritis, lymphoma, and stomach cancers. Although specific drugs are available to treat the bacterium's harmful effects, there is an urgent need to develop a preventive or therapeutic vaccine. Therefore, the current study aims to create a multi-epitope vaccine against H. pylori using lipid nanoparticles. Five epitopes from five target proteins of H. pylori, namely, Urease, CagA, HopE, SabA, and BabA, were used. Immunogenicity, MHC (Major Histocompatibility Complex) bonding, allergenicity, toxicity, physicochemical analysis, and global population coverage of the entire epitopes and final construct were carefully examined. The study involved using various bioinformatic web tools to accomplish the following tasks: modeling the three-dimensional structure of a set of epitopes and the final construct and docking them with Toll-Like Receptor 4 (TLR4). In the experimental phase, the final multi-epitope construct was synthesized using the solid phase method, and it was then enclosed in lipid nanoparticles. After synthesizing the construct, its loading, average size distribution, and nanoliposome shape were checked using Nanodrop at 280 nm, dynamic light scattering (DLS), and atomic force microscope (AFM). The designed vaccine has been confirmed to be non-toxic and anti-allergic. It can bind with different MHC alleles at a rate of 99.05%. The construct loading was determined to be about 91%, with an average size of 54 nm. Spherical shapes were also observed in the AFM images. Further laboratory tests are necessary to confirm the safety and immunogenicity of the multi-epitope vaccine.
Subject(s)
Bacterial Vaccines , Computational Biology , Helicobacter pylori , Nanoparticles , Helicobacter pylori/immunology , Nanoparticles/chemistry , Bacterial Vaccines/immunology , Bacterial Vaccines/chemistry , Computational Biology/methods , Humans , Bacterial Proteins/immunology , Bacterial Proteins/chemistry , Epitopes/immunology , Epitopes/chemistry , Molecular Docking Simulation , Antigens, Bacterial/immunology , Antigens, Bacterial/chemistry , Helicobacter Infections/prevention & control , Helicobacter Infections/immunology , Toll-Like Receptor 4/immunology , Urease/immunology , Urease/chemistry , Immunoinformatics , LiposomesABSTRACT
BACKGROUND: Theoretically, a rapid urease test (RUT) using a swab of the gastric wall (Swab-RUT) for Helicobacter pylori (H. pylori) is safe. However, the validity and utility of Swab-RUT remain unclear. Therefore, we assessed the validity and utility of Swab-RUT compared to RUT using mucosal forceps of the gastric wall (Forceps-RUT) and 13C-urea breath test (UBT). METHODS: This study was a multicenter prospective observational study. When the examinees were suspected of H. pylori infection during esophagogastroduodenoscopy, we performed Swab-RUT and Forceps-RUT continuously. When the examinees were not suspected of H. pylori infection, we performed Swab-RUT alone. We validated the status of H. pylori infection using UBT. RESULTS: Ninety-four examinees were enrolled from four institutions between May 2016 and December 2020 (median age [range], 56.5 [26-88] years). In this study, the sensitivity, specificity, and accuracy of Swab-RUT to UBT were 0.933 (95% confidence interval: 0.779-0.992), 0.922 (0.827-0.974), and 0.926 (0.853-0.970), respectively. The Kappa coefficient of Swab-RUT to UBT was 0.833, and that of Swab-RUT to forceps-RUT was 0.936. No complications were observed in this study. CONCLUSIONS: Swab-RUT is a valid examination for the status of H. pylori infection compared to the conventional Forceps-RUT.
Subject(s)
Breath Tests , Helicobacter Infections , Helicobacter pylori , Sensitivity and Specificity , Urease , Humans , Breath Tests/methods , Breath Tests/instrumentation , Helicobacter Infections/diagnosis , Helicobacter Infections/microbiology , Middle Aged , Prospective Studies , Urease/analysis , Urease/metabolism , Male , Female , Aged , Helicobacter pylori/isolation & purification , Helicobacter pylori/enzymology , Adult , Aged, 80 and over , Gastric Mucosa/microbiology , Endoscopy, Digestive System , Reproducibility of Results , Carbon Isotopes , Surgical Instruments/microbiologyABSTRACT
A novel and sensitive fluorescence ratiometric method is developed for urea detection based on the pH-sensitive response of two fluorescent carbon dot (CD) systems: R-CDs/methyl red (MR) and NIR-CDs/Cu2+. The sensing mechanism involves breaking down urea using the enzyme urease, releasing ammonia and increasing pH. At higher pH, the fluorescence of NIR-CDs is quenched due to the enhanced interaction with Cu2+, while the fluorescence of R-CDs is restored as the acidic MR converts to its basic form, removing the inner filter effect. The ratiometric signal (F608/F750) of the R-CDs/MR and NIR-CDs/Cu2+ intensities changed in response to the pH induced by urea hydrolysis, enabling selective and sensitive urea detection. Detailed spectroscopic and morphological investigations confirmed the fluorescence probe design and elucidated the sensing mechanism. The method exhibited excellent sensitivity (0.00028 mM LOD) and linearity range (0.001 - 8.0 mM) for urea detection, with successful application in milk samples for monitoring adulteration, demonstrating negligible interference and high recovery levels (96.5% to 101.0%). This ratiometric fluorescence approach offers a robust strategy for selective urea sensing in complicated matrices.
Subject(s)
Carbon , Copper , Fluorescent Dyes , Limit of Detection , Quantum Dots , Spectrometry, Fluorescence , Urea , Urease , Urea/analysis , Urea/chemistry , Urease/chemistry , Copper/chemistry , Carbon/chemistry , Hydrogen-Ion Concentration , Quantum Dots/chemistry , Fluorescent Dyes/chemistry , Spectrometry, Fluorescence/methods , Animals , Milk/chemistry , Azo Compounds/chemistry , Food Contamination/analysisABSTRACT
BACKGROUND: Rhododendron delavayi is a natural shrub that is distributed at different elevations in the karst region of Bijie, China, and that has an important role in preventing land degradation in this region. In this study, we determined the soil mineral element contents and soil enzyme activities. The composition of the soil bacterial community of R. delavayi at three elevations (1448 m, 1643 m, and 1821 m) was analyzed by high-throughput sequencing, and the interrelationships among the soil bacterial communities, mineral elements, and enzyme activities were determined. RESULTS: The Shannon index of the soil bacterial community increased and then decreased with increasing elevation and was highest at 1643 m. Elevations increased the number of total nodes and edges of the soil bacterial community network, and more positive correlations at 1821 m suggested stronger intraspecific cooperation. Acidobacteria, Actinobacteria and Proteobacteria were the dominant phyla at all three elevations. The Mantel test and correlation analysis showed that Fe and soil urease significantly affected bacterial communities at 1448 m; interestingly, Chloroflexi was positively related to soil urease at 1448 m, and Actinobacteria was positively correlated with Ni and Zn at 1821 m. Fe and soil urease significantly influenced the bacterial communities at lower elevations, and high elevation (1821 m) enhanced the positive interactions of the soil bacteria, which might be a strategy for R. delavayi to adapt to high elevation environments. CONCLUSION: Elevation significantly influenced the composition of soil bacterial communities by affecting the content of soil mineral elements and soil enzyme activity.
Subject(s)
Bacteria , Forests , Rhododendron , Soil Microbiology , Soil , Soil/chemistry , Rhododendron/microbiology , China , Bacteria/classification , Bacteria/genetics , Bacteria/enzymology , Bacteria/isolation & purification , Metals/analysis , Actinobacteria/genetics , Actinobacteria/enzymology , Actinobacteria/isolation & purification , Actinobacteria/classification , Microbiota , Urease/metabolism , Acidobacteria/genetics , Acidobacteria/isolation & purification , Acidobacteria/enzymology , Acidobacteria/classification , RNA, Ribosomal, 16S/genetics , Phylogeny , High-Throughput Nucleotide SequencingABSTRACT
Emerging evidence suggests differential antagonism of lactic acid-producing bacteria (LAB) to Helicobacter pylori, posing challenges to human health and food safety due to unclear mechanisms. This study assessed 21 LAB strains from various sources on H. pylori growth, urease activity, and coaggregation. Composite scoring revealed that Latilactobacillus sakei LZ217, derived from fresh milk, demonstrates strong inhibitory effects on both H. pylori growth and urease activity. L. sakei LZ217 significantly reduced H. pylori adherence of gastric cells in vitro, with inhibition ratios of 47.62%. Furthermore, in vivo results showed that L. sakei LZ217 alleviated H. pylori-induced gastric mucosa damage and inflammation in mice. Metabolomic exploration revealed metabolic perturbations in H. pylori induced by L. sakei LZ217, including reduced amino acid levels (e.g., isoleucine, leucine, glutamate, aspartate, and phenylalanine) and impaired carbohydrate and nucleotide synthesis, contributing to the suppression of ureA (28.30%), ureE (84.88%), and ureF (59.59%) expressions in H. pylori. This study underscores the efficacy of LAB against H. pylori and highlights metabolic pathways as promising targets for future interventions against H. pylori growth and colonization.
Subject(s)
Gastric Mucosa , Helicobacter Infections , Helicobacter pylori , Urease , Urease/metabolism , Animals , Helicobacter Infections/microbiology , Gastric Mucosa/microbiology , Gastric Mucosa/metabolism , Mice , Humans , Bacterial Adhesion , Female , Probiotics , MaleABSTRACT
We present the development and validation of an impedance-based urine osmometer for accurate and portable measurement of urine osmolality. The urine osmolality of a urine sample can be estimated by determining the concentrations of the conductive solutes and urea, which make up approximately 94% of the urine composition. Our method utilizes impedance measurements to determine the conductive solutes and urea after hydrolysis with urease enzyme. We built an impedance model using sodium chloride (NaCl) and urea at various known concentrations. In this work, we validated the accuracy of the impedance-based urine osmometer by developing a proof-of-concept first prototype and an integrated urine dipstick second prototype, where both prototypes exhibit an average accuracy of 95.5 ± 2.4% and 89.9 ± 9.1%, respectively in comparison to a clinical freezing point osmometer in the hospital laboratory. While the integrated dipstick design exhibited a slightly lower accuracy than the first prototype, it eliminated the need for pre-mixing or manual pipetting. Impedance calibration curves for conductive and non-conductive solutes consistently yielded results for NaCl but underscored challenges in achieving uniform urease enzyme coating on the dipstick. We also investigated the impact of storing urine at room temperature for 24 hours, demonstrating negligible differences in osmolality values. Overall, our impedance-based urine osmometer presents a promising tool for point-of-care urine osmolality measurements, addressing the demand for a portable, accurate, and user-friendly device with potential applications in clinical and home settings.
Subject(s)
Electric Impedance , Urea , Urease , Urea/urine , Urea/chemistry , Osmolar Concentration , Hydrolysis , Humans , Urease/metabolism , Urease/chemistry , Urinalysis/instrumentation , Equipment DesignABSTRACT
Enzyme-induced carbonate precipitation (EICP) has been studied in remediation of heavy metal contaminated water or soil in recent years. This paper aims to investigate the immobilization mechanism of Zn2+, Ni2+, and Cr(VI) in contaminated sand, as well as strength enhancement of sand specimens by using EICP method with crude sword bean urease extracts. A series of liquid batch tests and artificially contaminated sand remediation experiments were conducted to explore the heavy metal immobilization efficacy and mechanisms. Results showed that the urea hydrolysis completion efficiency decreased as the Ca2+ concentration increased and the heavy metal immobilization percentage increased with the concentration of Ca2+ and treatment cycles in contaminated sand. After four treatment cycles with 0.5 mol/L Ca2+ added, the immobilization percentage of Zn2+, Ni2+, and Cr(VI) were 99.99 %, 86.38 %, and 75.18 %, respectively. The microscale analysis results presented that carbonate precipitates and metallic oxide such as CaCO3, ZnCO3, NiCO3, Zn(OH)2, and CrO(OH) were generated in liquid batch tests and sand remediation experiments. The SEM-EDS and FTIR results also showed that organic molecules and CaCO3 may adsorb or complex heavy metal ions. Thus, the immobilization mechanism of EICP method with crude sword bean urease can be considered as biomineralization, as well as adsorption and complexation by organic matter and calcium carbonate. The unconfined compressive strength of EICP-treated contaminated sand specimens demonstrated a positive correlation with the increased generation of carbonate precipitates, being up to 306 kPa after four treatment cycles with shear failure mode. Crude sword bean urease with 0.5 mol/L Ca2+ added is recommended to immobilize multiple heavy metal ions and enhance soil strength.
Subject(s)
Environmental Restoration and Remediation , Metals, Heavy , Sand , Soil Pollutants , Urease , Environmental Restoration and Remediation/methods , Carbonates/chemistry , Compressive Strength , Chemical PrecipitationABSTRACT
The clearance of urea poses a formidable challenge, and its excessive accumulation can cause various renal diseases. Urease demonstrates remarkable efficacy in eliminating urea, but cannot be reused. This study aimed to develop a composite vector system comprising microcrystalline cellulose (MCC) immobilized with urease and metal-organic framework (MOF) UiO-66-NH2, denoted as MCC@UiO/U, through the dynamic defect generation strategy. By utilizing competitive coordination, effective immobilization of urease into MCC@UiO was achieved for efficient urea removal. Within 2 h, the urea removal efficiency could reach up to 1500 mg/g, surpassing an 80% clearance rate. Furthermore, an 80% clearance rate can also be attained in peritoneal dialyzate from patients. MCC@UiO/U also exhibits an exceptional bioactivity even after undergoing 5 cycles of perfusion, demonstrating remarkable stability and biocompatibility. This innovative approach and methodology provide a novel avenue and a wide range of immobilized enzyme vectors for clinical urea removal and treatment of kidney diseases, presenting immense potential for future clinical applications.
Subject(s)
Cellulose , Enzymes, Immobilized , Metal-Organic Frameworks , Urea , Urease , Urease/chemistry , Urease/metabolism , Urea/chemistry , Enzymes, Immobilized/chemistry , Enzymes, Immobilized/metabolism , Cellulose/chemistry , Metal-Organic Frameworks/chemistry , HumansABSTRACT
We tested the ability of wood distillate (WD) to interact with urea in agricultural soil. WD is a sustainable material that has been addressed as a promising alternative to synthetic soil corroborants. However, there is little information about the effect of WD on the nitrogen cycle. In this study, soils with different amounts of WD and with/without urea were tested for ammonium, urease, nitrate/nitrite, and potential nitrification activity at different points in a 30 day time frame. High concentrations of WD (1-2%) inhibited the hydrolysis of urea and the oxidation of ammonium to nitrate. Thermal desorption coupled to GC-MS and high-performance liquid chromatography-tandem mass spectrometry characterization allowed us to reveal that WD-urea interactions mainly involve lignin-derived compounds in the distillate, such as catechol, resorcinol, and syringol. This study provides the first evidence of a strong interaction between WD and urea in soil that could be used to develop slow-release fertilizers.
Subject(s)
Fertilizers , Wood , Wood/chemistry , Urea/chemistry , Urea/metabolism , Urease/metabolism , Chromatography, High Pressure Liquid , Gas Chromatography-Mass Spectrometry , Nitrification , Ammonium Compounds/chemistryABSTRACT
The synergistic remediation of heavy metal-contaminated soil by functional strains and biochar has been widely studied. However, the mechanisms by which urease-producing bacteria combine with pig manure biochar (PMB) to immobilize Cd and inhibit Cd absorption in vegetables are still unclear. In our study, the effects and mechanisms of PMB combined with the urease-producing bacterium TJ6 (TJ6 + PMB) on Cd adsorption were explored. The effects of TJ6 + PMB on the Cd content and pH of the leachate were also studied through a 56-day soil leaching experiment. Moreover, the effects of the complexes on Cd absorption and microbial mechanisms in lettuce were explored through pot experiments. The results showed that PMB provided strain TJ6 with a greater ability to adsorb Cd, inducing the generation of CdS and CdCO3, and thereby reducing the Cd content (71.1%) and increasing the pH and urease activity in the culture medium. TJ6 + PMB improved lettuce dry weight and reduced Cd absorption. These positive effects were likely due to (1) TJ6 + PMB increased the organic matter and NH4+ contents, (2) TJ6 + PMB transformed available Cd into residual Cd and decreased the Cd content in the leachate, and (3) TJ6 + PMB altered the structure of the rhizosphere bacterial and fungal communities in lettuce, increasing the relative abundances of Stachybotrys, Agrocybe, Gaiellales, and Gemmatimonas. These genera can promote plant growth, decompose organic matter, and release phosphorus. Interestingly, the fungal communities were more sensitive to the addition of TJ6 and PMB, which play important roles in the decomposition of organic matter and immobilization of Cd. In conclusion, this study revealed the mechanism by which urease-producing bacteria combined with pig manure biochar immobilize Cd and provided a theoretical basis for safe pig manure return to Cd-polluted farmland. This study also provides technical approaches and bacterial resources for the remediation of heavy metal-contaminated soil.
Subject(s)
Cadmium , Charcoal , Lactuca , Manure , Soil Pollutants , Urease , Cadmium/metabolism , Charcoal/chemistry , Animals , Urease/metabolism , Swine , Soil Pollutants/metabolism , Bacteria/metabolism , Soil/chemistryABSTRACT
Sodium ions and protons regulate various fundamental processes at the cell and tissue levels across all biological kingdoms. It is therefore pivotal for bioelectronic devices, such as biosensors and biotransducers, to control the transport of these ions through biological membranes. Our study explores the regulation of proton and sodium concentrations by integrating an Na+-type ATP synthase, a glucose dehydrogenase (GDH), and a urease into a multienzyme logic system. This system is designed to operate using various chemical control input signals, while the output current corresponds to the local change in proton or sodium concentrations. Therein, a H+ and Na+ biotransducer was integrated to fulfill the roles of signal transducers for the monitoring and simultaneous control of Na+ and H+ levels, respectively. To increase the proton concentration at the output, we utilized GDH driven by the inputs of glucose and nicotinamide adenine dinucleotide (NAD+), while recorded the signal change from the biotransducer, together acting as an AND enzyme logic gate. On the contrary, we introduced urease enzyme which hydrolyzed urea to control the decrease in proton concentration, serving as a NOT gate and reset. By integrating these two enzyme logic gates we formed a simple multienzyme logic system for the control of proton concentrations. Furthermore, we also demonstrate a more complex, Na+-type ATP synthase-urease multienzyme logic system, controlled by the two different inputs of ADP and urea. By monitoring the voltage of the peak current as the output signal, this logic system acts as an AND enzyme logic gate. This study explores how multienzyme logic systems can modulate biologically important ion concentrations, opening the door toward advanced biological on-demand control of a variety of bioelectronic enzyme-based devices, such as biosensors and biotransducers.
Subject(s)
Glucose 1-Dehydrogenase , Sodium , Sodium/metabolism , Sodium/chemistry , Glucose 1-Dehydrogenase/metabolism , Glucose 1-Dehydrogenase/chemistry , Urease/metabolism , Urease/chemistry , Protons , Glucose/metabolism , Biosensing Techniques/methods , NAD/metabolism , NAD/chemistryABSTRACT
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.
Subject(s)
Acid Phosphatase , Benzhydryl Compounds , Phenols , Soil Pollutants , Soil , Zea mays , Phenols/chemistry , Benzhydryl Compounds/chemistry , Soil Pollutants/chemistry , Zea mays/chemistry , Soil/chemistry , Acid Phosphatase/metabolism , Arylsulfatases/metabolism , Alkaline Phosphatase/metabolism , Zeolites/chemistry , Oxidoreductases/metabolism , Urease/metabolism , Catalase/metabolism , Biodegradation, Environmental , Magnesium Silicates/chemistry , Starch/chemistry , beta-Glucosidase/metabolism , Composting/methodsABSTRACT
In order to evaluate the effects of continuous cropping of millet on soil nutrients and soil enzyme activities, the present study was based on four treatments of 2 years of continuous cropping (T1), 3 years of continuous cropping (T2), 4 years of continuous cropping (T3) and rotational cropping (CK), based on 4 years of no fertilizer positioning experiments, and the soil nutrients, soil enzyme activities and millets yields were determined, respectively. The results showed that with the increase of continuous cropping years, the millet yield decreased and was significantly lower than that of rotating with legume crops, and compared with CK, the yields of T1, T2 and T3 treatments were reduced by 8.92%, 13.73% and 37.60%, respectively; the soil nitrogen and phosphorus contents were reduced, the quick-acting potassium content did not change obviously, and the soil pH was increased; Soil urease, alkaline phosphatase, sucrase and catalase activities generally showed a decreasing trend and the decrease was more significant with the increase in the number of years of continuous cropping. Therefore, in order to maintain the soil fertility and increase the millet yield, it is necessary to practice crop rotation and stubble reversal between millets and leguminous crops such as kidney beans, and to apply certain fertilizers.
Subject(s)
Crops, Agricultural , Fertilizers , Millets , Nitrogen , Soil , Soil/chemistry , Nitrogen/analysis , Nitrogen/metabolism , Crops, Agricultural/growth & development , Fertilizers/analysis , Phosphorus/analysis , Phosphorus/metabolism , Nutrients/analysis , Agriculture/methods , Crop Production/methods , Potassium/analysis , Potassium/metabolism , Alkaline Phosphatase/metabolism , Hydrogen-Ion Concentration , Urease/metabolism , Sucrase/metabolismABSTRACT
Autonomous nanorobots represent an advanced tool for precision therapy to improve therapeutic efficacy. However, current nanorobotic designs primarily rely on inorganic materials with compromised biocompatibility and limited biological functions. Here, we introduce enzyme-powered bacterial outer membrane vesicle (OMV) nanorobots. The immobilized urease on the OMV membrane catalyzes the decomposition of bioavailable urea, generating effective propulsion for nanorobots. This OMV nanorobot preserves the unique features of OMVs, including intrinsic biocompatibility, immunogenicity, versatile surface bioengineering for desired biofunctionalities, capability of cargo loading and protection. We present OMV-based nanorobots designed for effective tumor therapy by leveraging the membrane properties of OMVs. These involve surface bioengineering of robotic body with cell-penetrating peptide for tumor targeting and penetration, which is further enhanced by active propulsion of nanorobots. Additionally, OMV nanorobots can effectively safeguard the loaded gene silencing tool, small interfering RNA (siRNA), from enzymatic degradation. Through systematic in vitro and in vivo studies using a rodent model, we demonstrate that these OMV nanorobots substantially enhanced siRNA delivery and immune stimulation, resulting in the utmost effectiveness in tumor suppression when juxtaposed with static groups, particularly evident in the orthotopic bladder tumor model. This OMV nanorobot opens an inspiring avenue to design advanced medical robots with expanded versatility and adaptability, broadening their operation scope in practical biomedical domains.
Subject(s)
Bacterial Outer Membrane , Animals , Humans , Bacterial Outer Membrane/metabolism , Mice , Robotics/methods , Urease/metabolism , RNA, Small Interfering/genetics , RNA, Small Interfering/metabolism , Enzymes, Immobilized/chemistry , Enzymes, Immobilized/metabolismABSTRACT
Objective: The study aimed to detect the presence of Helicobacter pylori infection in children using two investigative methods: the rapid urease test and histological methods. It also examined the relationship between socioeconomic status and Helicobacter pylori infection. Design: This was a cross-sectional study conducted in the paediatric theatre at Korle Bu Teaching Hospital in Accra, Ghana. Participants: Children who were scheduled for upper gastrointestinal endoscopy were recruited into the study. Main outcome measures: The presence of Helicobacter pylori in gastric biopsies was measured using a rapid urease test and histology. Results: Seventy-three children aged 2 years to 16 years were seen during the period. Both tests were positive at the same time in 36 (49.3%) out of the 73 children (p<0.0001). The positivity rates for the rapid urease test and histology were 57.5% and 53.4 %, respectively. Significant predictors of the histology presence of H. pylori were a large household size of at least 6 members (AOR: 4.03; p<0.013) and the presence of pets at home (AOR: 3.23; p<0.044). Conclusions: Substantial agreement was found between the rapid urease test and histology examination of gastric biopsies for the presence of H. pylori. Children from large households and those with pets at home appear to have increased odds of having H. pylori infection of the gastric mucosa. Funding: None declared.
Subject(s)
Helicobacter Infections , Helicobacter pylori , Urease , Humans , Helicobacter Infections/diagnosis , Helicobacter pylori/isolation & purification , Child , Cross-Sectional Studies , Male , Urease/analysis , Female , Child, Preschool , Adolescent , Ghana/epidemiology , Biopsy , Socioeconomic Factors , Gastric Mucosa/pathology , Gastric Mucosa/microbiologyABSTRACT
Microbially induced carbonate precipitation (MICP) immobilizes toxic metals and reduces their bioavailability in aqueous systems. However, its application in the treatment of acid mine drainage (AMD) is poorly understood. In this study, the genomes of Sporosarcina sp. UB5 and UB10 were sequenced. Urease, carbonic anhydrases, and metal resistance genes were identified and enzymatic assays were performed for their validation. The geochemical mechanism of precipitation in AMD was elucidated through geo-mineralogical analysis. Sporosarcina sp. UB5 was shown to be a new genomospecies, with an average nucleotide identity < 95 % (ANI) and DNA-DNA hybridization < 70 % (DDH) whereas UB10 is close to S. pasteurii. UB5 contained two urease operons, whereas only one was identified in UB10. The ureolytic activities of UB5 and UB10 were 122.67 ± 15.74 and 131.70 ± 14.35 mM NH4+ min-1, respectively. Both strains feature several carbonic anhydrases of the α, ß, or γ families, which catalyzed the precipitation of CaCO3. Only Sporosarcina sp. UB5 was able to immobilize metals and neutralize AMD. Geo-mineralogical analyses revealed that UB5 directly immobilized Fe (1-23 %), Mn (0.65-1.33 %) and Zn (0.8-3 %) in AMD via MICP and indirectly through adsorption to calcite and binding to bacterial cell walls. The MICP-treated AMD exhibited high removal rates (>67 %) for Ag, Al, As, Ca, Cd, Co, Cu, Fe, Mn, Pb, and Zn, and a removal rate of 15 % for Mg. This study provides new insights into the MICP process and its applications to AMD treatment using autochthonous strains.
Subject(s)
Mining , Sporosarcina , Urease , Sporosarcina/genetics , Sporosarcina/metabolism , Urease/metabolism , Chemical Precipitation , Carbonates/chemistry , Carbonic Anhydrases/metabolism , Carbonic Anhydrases/genetics , Water Pollutants, Chemical/metabolism , Water Pollutants, Chemical/chemistryABSTRACT
The biomimetic enzyme cascade system plays a key role in biosensing as a sophisticated signal transduction and amplification strategy. However, constructing a regulated enzyme cascade sensing system remains challenging due to the mismatch of multiple enzyme activities and poor stability. Herein, we design an efficient dual-enhanced enzyme cascade hybrid system (UFD-DEC) containing DNA-controlled nanozymes (Fe-cdDNA) and enzyme (urease) via combining the electrostatic contact effect with the hydrogel-directed confinement effect. Precise modulation of Fe-cdDNA nanozyme by DNA offers a means to control its catalytic efficiency. This regulated UFD-DEC system accelerates the reaction rate and provides remarkable stability compared with the free enzyme system. Benefiting from the plasticity properties of hydrogels, a "lab-in-a-tube" platform was constructed by encapsulating UFD-DEC in a microcentrifuge tube. Such a UFD-DEC-based hydrogel tube exhibits sufficient adaptability to profile urea when used in conjunction with a smartphone-assisted image processing algorithm, which on-site delivers urea information with a detection limit of 0.12 mmol L-1. This customizable and inexpensive miniaturized biosensor platform for monitoring urea may facilitate point-of-care testing applications.
Subject(s)
Biosensing Techniques , Hydrogels , Limit of Detection , Urease , Biosensing Techniques/methods , Hydrogels/chemistry , Urease/chemistry , Urea/analysis , Urea/chemistry , DNA, Catalytic/chemistry , DNA/chemistryABSTRACT
Urinary tract infection caused by Klebsiella, Proteus and Streptococcus is a urease dependent process, hence treatment of these infections by antibacterial compounds lies in inhibition of their virulence factors. The crude methanolic extracts derived from sumac fruit, pomegranate peel and Indian almond leaves were separated into anthocyanin and non-anthocyanin fractions using solid phase cartridges. The inhibitory effect of these fractions was determined on the growth of urease producing species and jack bean urease activity. Known compounds in the fractions were also docked with ureases of different biological origins viz. K. pneumoniae (PDB ID: 8HCN), K. aerogenes (PDB ID: 2KAU), Helicobacter pylori (PDB ID:8HC1)and Canavalia ensiformis (jack bean) (PDB ID: 3LA4) to determine their binding affinities and interaction with the enzyme. All the fractions showed significant inhibition growth for P. mirabilis, S. epidermidis and K.pneumoniae. Among the samples, sumac showed greatest inhibition against all (MIC 6-25 mg.mL-1) while among the fractions, anthocyanin was found to be most active (MIC 6-12 mg/mL). Likewise, all fractions inhibited urease with lowest ICs50 shown by sumac fractions (21-116 µg.mL-1). Out of 39 compounds docked, 27 showed interaction with movable flaps and/or active site of ureases which explains their mode of inhibition.
Subject(s)
Polyphenols , Pomegranate , Prunus dulcis , Urease , Urease/antagonists & inhibitors , Urease/metabolism , Polyphenols/pharmacology , Polyphenols/chemistry , Pomegranate/chemistry , Prunus dulcis/chemistry , Canavalia/enzymology , Plant Extracts/pharmacology , Plant Extracts/chemistry , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/chemistry , Enzyme Inhibitors/pharmacology , Enzyme Inhibitors/chemistry , Microbial Sensitivity Tests , Molecular Docking Simulation , Bacteria/drug effects , Bacteria/enzymologyABSTRACT
Microbially induced carbonate precipitation (MICP) is a common biomineralization method, which is often used for remediation of heavy metal pollution such as hexavalent chromium (Cr(VI)) in recent years. Calcium sources are essential for the MICP process. This study investigated the potential of MICP technology for Cr(VI) remediation under the influence of three calcium sources (CaCl2, Ca(CH3COO)2, Ca(C6H11O7)2). The results indicated that CaCl2 was the most efficient in the mineralization of Cr(VI), and Ca(C6H11O7)2 could significantly promote Cr(VI) reduction. The addition of different calcium sources all promoted the urease activity of Sporosarcina saromensis W5, in which the CaCl2 group showed higher urease activity at the same Ca2+ concentration. Besides, with CaCl2, Ca(CH3COO)2 and Ca(C6H11O7)2 treatments, the final fraction of Cr species (Cr(VI), reduced Cr(III) and organic Cr(III)-complexes) were mainly converted to the carbonate-bound, cytoplasm and cell membrane state, respectively. Furthermore, the characterization results revealed that three calcium sources could co-precipitate with Cr species to produce Ca10Cr6O24(CO3), and calcite and vaterite were present in the CaCl2 and Ca(CH3COO)2 groups, while only calcite was present in the Ca(C6H11O7)2 group. Overall, this study contributes to the optimization of MICP-mediated remediation of heavy metal contaminated soil. CaCl2 was the more suitable calcium source than the other two for the application of MICP technology in the Cr(VI) reduction and mineralization.
Subject(s)
Calcium , Carbonates , Chromium , Sporosarcina , Chromium/metabolism , Chromium/chemistry , Calcium/metabolism , Sporosarcina/metabolism , Carbonates/chemistry , Carbonates/metabolism , Chemical Precipitation , Urease/metabolism , Biodegradation, Environmental , Calcium Chloride/chemistry , Soil Pollutants/metabolismABSTRACT
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.