Revolutionizing elderly care: Building a healthier aging society through innovative long-term care systems and assessing the long-term care acceptance model.

AIM: With a growing elderly population, the demand for caregivers is increasing in Khon Kaen, Thailand, with approximately 17 000 elderly residents. This growing number of older people and a shortage of caregivers could overload the healthcare system. METHODS: The present study involved 129 healthcare volunteers (caregivers for questionnaires study) and the collection of health data from 290 elderly residents from northeastern Thailand. After training, the volunteers assessed its usefulness through questionnaires. Tool reliability and statistical hypotheses were tested using stratified regression analysis (hierarchical regression) and multiple regression. RESULTS: The relative mean scores of perceived usefulness, perceived ease of use, attitude toward usage and behavioral intention to use technology were 4.51, 4.29, 4.44 and 4.41, respectively. In addition, perceived usefulness and user attitudes positively affected volunteers' willingness to use the system. CONCLUSION: The study was developed from the awareness of enhancing community quality and ecosystem through a long-term care system application. Analyzing external factors can enhance technology's future effectiveness. Geriatr Gerontol Int 2024; ••: ••-••.

Highly Selective Colorimetric Sensor of Mercury(II) Ions by Andrographolide-Stabilized Silver Nanoparticles in Water and Antibacterial Evaluation.

Silver nanoparticles (AgNPs) are well known for their exceptional properties and versatility in various applications. This study used andrographolide as a biochemical stabilizer to synthesize AgNPs (andro-AgNPs). The andro-AgNPs were characterized by using UV-vis spectroscopy, revealing a surface plasmon resonance peak at 440 nm. Fourier transform infrared spectroscopy was also used to confirm the presence of AgNPs. Transmission electron microscopy was used to investigate the morphology of andro-AgNPs, which showed a spherical shape with an average diameter of 18.30 ± 5.57 nm (n = 205). Andro-AgNPs were utilized as a colorimetric sensor to detect mercury ions (Hg2+) in water, and the optimized detection conditions were evaluated using UV-vis spectroscopy with a linear range of 15-120 µM. The limit of detection and the limit of quantification for Hg2+ detection were found to be 11.15 and 37.15 µM, respectively. Furthermore, andro-AgNPs exhibited antibacterial properties against both Gram-positive (Staphylococcus aureus) and Gram-negative (Escherichia coli) bacteria. The results imply that andro-AgNPs hold promising potential for future biomedical applications.

The effect of edible bird's nests on the expression of MHC-II and costimulatory molecules of C57BL/6 mouse splenocytes.

The glutinous nest that builds by the saliva secretion of swiftlet is recognizable as an edible bird's nest (EBN). It enriched a medicinal value and was regarded as supplementary food that exerts various beneficial health effects, especially immune boosters. This study's objective was to determine the impact of EBN on the expression of MHC-II and costimulatory molecules (CD86 and CD80) related to the initiation of T-cell activation. Both rEBN and pEBN samples were prepared with simulated gastrointestinal digestion for enhancing the bioaccessibility of bioactive compounds. Our result showed that digested EBN samples slightly influence the upregulation of MHC-II, CD86, and CD80 in gene expression of LPS-stimulated Raw 264.7 cells. The concern of endotoxin contamination in EBN samples, which may cause a false-positive result, was measured by quantitative PCR. We found that the inflammatory genes (IL-1ß and TNF-α) were not induced by EBN treatments. Moreover, cell surface protein expression in splenocytes treated with EBN was assessed using flow cytometric analysis. Digested EBN samples demonstrated their capacity to promote the elevation of MHC-II, CD86, and CD80 cell surface protein expression. Finally, the digested-EBN-treated splenocytes only exhibited a specific response in the T-cells population. Thus, EBN is a source of the bioactive compound that has been proposed to exert a role in the stimulation of both MHC-II and costimulatory molecules for TCR/pMHC-II interaction leading to T-cell activation.

A Shear-Thinning, Self-Healing, Dual-Cross Linked Hydrogel Based on Gelatin/Vanillin/Fe3+ /AGP-AgNPs: Synthesis, Antibacterial, and Wound-Healing Assessment.

A shear-thinning and self-healing hydrogel based on a gelatin biopolymer is synthesized using vanillin and Fe3+ as dual crosslinking agents. Rheological studies indicate the formation of a strong gel found to be injectable and exhibit rapid self-healing (within 10 min). The hydrogels also exhibited a high degree of swelling, suggesting potential as wound dressings since the absorption of large amounts of wound exudate, and optimum moisture levels, lead to accelerated wound healing. Andrographolide, an anti-inflammatory natural product is used to fabricate silver nanoparticles, which are characterized and composited with the fabricated hydrogels to imbue them with anti-microbial activity. The nanoparticle/hydrogel composites exhibit activity against Escherichia coli, Staphylococcus aureus, and Burkholderia pseudomallei, the pathogen that causes melioidosis, a serious but neglected disease affecting southeast Asia and northern Australia. Finally, the nanoparticle/hydrogel composites are shown to enhance wound closure in animal models compared to the hydrogel alone, confirming that these hydrogel composites hold great potential in the biomedical field.

Gingerol extract-stabilized silver nanoparticles and their applications: colorimetric and machine learning-based sensing of Hg(ii) and antibacterial properties.

This study focused on synthesizing ginger-stabilized silver nanoparticles (Gin-AgNPs) using a more eco-friendly method that utilized AgNO3 and natural ginger solution. These nanoparticles underwent a color change from yellow to colorless when exposed to Hg2+, enabling the detection of Hg2+ in tap water. The colorimetric sensor had good sensitivity, with a limit of detection (LOD) of 1.46 µM and a limit of quantitation (LOQ) of 3.04 µM. Importantly, the sensor operated accurately without being affected by various other metal ions. To enhance its performance, a machine learning approach was employed and achieved accuracy ranging from 0% to 14.66% when trained with images of Gin-AgNP solutions containing different Hg2+ concentrations. Furthermore, the Gin-AgNPs and Gin-AgNPs hydrogels exhibited antibacterial effects against both Gram-negative and Gram-positive bacteria, indicating potential future applications in the detection of Hg2+ and in wound healing.

Electrochemical biosensor based on cellulose nanofibers/graphene oxide and acetylcholinesterase for the detection of chlorpyrifos pesticide in water and fruit juice.

In this work, cellulose nanofibers and graphene oxide are used to fabricate a simple and reliable electrochemical biosensor, based on acetylcholinesterase (AChE) for the detection of highly dangerous organophosphates (OPs), utilizing chlorpyrifos as a representative sample. AChE is an enzyme that is essential for neurotransmission and catalyzes the conversion of acetylcholine (ATCh) into thiocholine and acetic acid. The pesticide used in this work, chlorpyrifos, inhibits the catalytic activity of AChE on ATCh, and this inhibition can be measured using square wave voltammetry (SWV). Utilizing a process of surface modification, layers of cellulose nanofibers, graphene oxide, a chitosan-graphene oxide composite, and acetylcholinesterase (AChE/CS-GO/GO/CNFs) were immobilized on a screen-printed carbon electrode (SPCE). The modified SPCE working electrode was characterized using scanning electron microscopy and graphene oxide trapped in the cellulose nanofibers was found to increase the sensitivity of the biosensor. The modified biosensor demonstrated good performance for detection of chlorpyrifos over a linear range of 25-1000 nM under optimum conditions, and the limits of detection and quantification were 2.2 nM and 73 nM, respectively. Our sophisticated technique might offer a more precise, straightforward, quick, and environmentally friendly way to assess chlorpyrifos contamination in water and juice samples.

Unveiling the Potent Antiviral and Antioxidant Activities of an Aqueous Extract from Caesalpinia mimosoides Lamk: Cheminformatics and Molecular Docking Approaches.

Our group previously demonstrated that Caesalpinia mimosoides Lamk exhibits many profound biological properties, including anticancer, antibacterial, and antioxidant activities. However, its antiviral activity has not yet been investigated. Here, the aqueous extract of C. mimosoides was prepared from the aerial parts (leaves, stalks, and trunks) to see whether it exerts anti-influenza (H1N1) effects and to reduce the organic solvents consumed during extraction, making it a desirable approach for the large-scale production for medical uses. Our plant extract was quantified to contain 7 g of gallic acid (GA) per 100 g of a dry sample, as determined using HPLC analysis. It also exerts potent antioxidant activities comparable to those of authentic GA. According to untargeted metabolomics (UPLC-ESI(-)-QTOF-MS/MS) with the aid of cheminformatics tools (MetFrag (version 2.1), SIRIUS (version 5.8.3), CSI:FingerID (version 4.8), and CANOPUS), the major metabolite was best annotated as "gallic acid", phenolics (e.g., quinic acid, shikimic acid, and protocatechuic acid), sugar derivatives, and dicarboxylic acids were deduced from this plant species for the first time. The aqueous plant extract efficiently inhibited an influenza A (H1N1) virus infection of MDCK cells with an IC50 of 5.14 µg/mL. Of equal importance, hemolytic activity was absent for this plant extract, signifying its applicability as a safe antiviral agent. Molecular docking suggested that GA interacts with conserved residues (e.g., Arg152 and Asp151) located in the catalytic inner shell of the viral neuraminidase (NA), sharing the same pocket as those of anti-neuraminidase drugs, such as laninamivir and oseltamivir. Additionally, other metabolites were also found to potentially interact with the active site and the hydrophobic 430-cavity of the viral surface protein, suggesting a possibly synergistic effect of various phytochemicals. Therefore, the C. mimosoides aqueous extract may be a good candidate for coping with increasing influenza virus resistance to existing antivirals.

TA-AgNPs/Alginate Hydrogel and Its Potential Application as a Promising Antibiofilm Material against Polymicrobial Wound Biofilms Using a Unique Biofilm Flow Model.

The presence of biofilm within a chronic wound may delay the healing process. Thus, control of biofilm formation and providing bactericidal effect are crucial factors for wound healing management. Alginate-based nanocomposite hydrogels have been suggested as dressing materials for wound treatment, which are employed as a biocompatible matrix. Therefore, in this study, we aimed to develop a biocompatible antimicrobial wound dressing containing AgNPs and demonstrate its efficacy against polymicrobial wound biofilms by using a biofilm flow device to simulate a chronic infected, exuding wound and specific wound environment. The results from agar well diffusion, the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) assays showed that TA-AgNPs exhibited antibacterial activity against wound pathogens. Additionally, the Minimum Biofilm Eradication Concentration assay (MBEC) demonstrated it could impair biofilm formation. Importantly, our TA-AgNPs/Alginate hydrogel clearly showed antibacterial activities against Streptococcus pyogenes, Staphylococcus aureus and Pseudomonas aeruginosa. Furthermore, we used the biofilm flow device to test the topical antimicrobial hydrogel against a three-species biofilm. We found that TA-AgNPs/Alginate hydrogel significantly showed a 3-4 log reduction in bacterial numbers when applied with multiple doses at 24 h intervals, and was especially effective against the chronic wound pathogen P. aeruginosa. This work highlighted that the TA-AgNPs/Alginate hydrogel is a promising material for treating complex wound biofilms.

Andrographolide stabilized-silver nanoparticles overcome ceftazidime-resistant Burkholderia pseudomallei: study of antimicrobial activity and mode of action.

Burkholderia pseudomallei (B. pseudomallei) is a Gram-negative pathogen that causes melioidosis, a deadly but neglected tropical disease. B. pseudomallei is intrinsically resistant to a growing list of antibiotics, and alternative antimicrobial agents are being sought with urgency. In this study, we synthesize andrographolide-stabilized silver nanoparticles (andro-AgNPs, spherically shaped with 16 nm average diameter) that show excellent antimicrobial activity against B. pseudomallei, including ceftazidime-resistant strains, being 1-3 orders of magnitude more effective than ceftazidime and 1-2 orders of magnitude more effective than other green-synthesized AgNPs. The andro-AgNPs are meanwhile non-toxic to mammalian cell lines. The mode of action of Andro-AgNPs toward B. pseudomallei is unraveled by killing kinetics, membrane neutralization, silver ions (Ag+) release, reactive oxygen species (ROS) induction, membrane integrity, and cell morphology change studies. The antimicrobial activity and mode of action of andro-AgNPs against B. pseudomallei reported here may pave the way to alternative treatments for melioidosis.

Anisotropic Silver Nanoparticles Gel Exhibits Antibacterial Action and Reduced Scar Formation on Wounds Contaminated with Methicillin-Resistant Staphylococcus pseudintermedius (MRSP) in a Mice Model.

Staphylococcus pseudintermedius (S. pseudintermedius) infected wounds can cause seriously delayed wound healing processes in animals. Antimicrobial agents that have antimicrobial and wound healing efficacy have become an essential tool for overcoming this problem. In our previous study, anisotropic AgNPs have been reported to have antimicrobial efficiency against animal and human pathogens, and could be suitable as antimicrobial agents for infected wounds. Here, antimicrobial and wound healing activities of anisotropic AgNPs gels were assessed in vivo. BALB/cAJcl mice wounds were infected by Methicillin-resistant Staphylococcus pseudintermedius (MRSP). Then, antibacterial and wound healing activities were evaluated by bacterial cell count, wound contraction, digital capture, and histology. The results show that anisotropic AgNPs gels could eliminate all bacterial cell infected wounds within 7 days, the same as povidone iodine. Wound healing activity was evaluated by wound contraction (%). The results showed 100% wound contraction in groups treated with anisotropic AgNPs gels within 14 days that was not significantly different from povidone iodine and control gel without AgNPs. However, the digital capture of wounds on day 4 showed that anisotropic AgNPs gel prevented pus formation and reduced scar appearance within 21 days. The histology results exhibit improved collagen fiber alignment that supports scar disappearance. In conclusion, these results indicate that anisotropic AgNPs gels are suitable for treating infected wounds. The gel is effective in eliminating bacteria that supports the natural process of wound repair and also causes reduced scar formation.

A Disposable Electrochemical Biosensor Based on Screen-Printed Carbon Electrodes Modified with Silver Nanowires/HPMC/Chitosan/Urease for the Detection of Mercury (II) in Water.

This work describes the facile preparation of a disposable electrochemical biosensor for the detection of Hg(II) in water by modifying the surface of a screen-printed carbon electrode (SPCE). The surface modification consists of the immobilization of a composite layer of silver nanowires, hydroxymethyl propyl cellulose, chitosan, and urease (AgNWs/HPMC/CS/Urease). The presence of the composite was confirmed by scanning electron microscopy (SEM) and its excellent conductivity, due chiefly to the electrical properties of silver nanowires, enhanced the sensitivity of the biosensor. Under optimum conditions, the modified SPCE biosensor showed excellent performance for the detection of Hg(II) ions, with an incubation time of 10 min and a linear sensitivity range of 5-25 µM. The limit of detection (LOD) and limit of quantitation (LOQ) were observed to be 3.94 µM and 6.50 µM, respectively. In addition, the disposable and portable biosensor exhibited excellent recoveries for the detection of Hg(II) ions in commercial drinking water samples (101.62-105.26%). The results are correlated with those obtained from inductively coupled plasma optical emission spectrometry (ICP-OES), indicating that our developed sensor is a reliable method for detection of Hg(II) in real water samples. The developed sensor device is a simple, effective, portable, low cost, and user-friendly platform for real-time detection of heavy metal ions in field measurements with potential for other biomedical applications in the future.

A Biological Study of Anisotropic Silver Nanoparticles and Their Antimicrobial Application for Topical Use.

The excessive use of antibiotics in both human and veterinary medicine has contributed to the development and rapid spread of drug resistance in bacteria. Silver nanoparticles (AgNPs) have become a tool of choice that can be used to treat these resistant bacteria. Several studies have shown that AgNPs have antibacterial and wound healing properties. In this study, we evaluated the biological activity of anisotropic AgNPs to develop an antimicrobial gel formulation for treating wound infections. We showed that some anisotropic AgNPs (S2) have an effective antibacterial activity against bacterial pathogens and low cytotoxicity to keratinocytes and fibroblasts in vitro. The MIC and MBC values were in the range of 2-32 µg/mL, and cytotoxicity had IC50 values of 68.20 ± 9.71 µg/mL and 68.65 ± 10.97 µg/mL against human keratinocyte and normal human dermal fibroblast cells, respectively. The anisotropic AgNPs (S2) were used as a gel component and tested for antibacterial activity, including long-term protection, compared with povidone iodine, a common antiseptic agent. The results show that the anisotropic AgNPs can inhibit the growth of most tested bacterial pathogens and provide protection longer than 48 h, whereas povidone iodine only inhibits the growth of some bacteria. This study suggests that anisotropic AgNPs could be used as an alternative antimicrobial agent for treating bacterial skin infection and as a wound healing formulation.

Silver Nanoparticles Enhance Antimicrobial Efficacy of Antibiotics and Restore That Efficacy against the Melioidosis Pathogen.

Melioidosis is an infectious disease caused by Gram-negative bacillus bacteria Burkholderia pseudomallei. Due to the emerging resistance of B. pseudomallei to antibiotics including ceftazidime (CAZ), the development of novel antibiotics and alternative modes of treatment has become an urgent issue. Here, we demonstrated an ability to synergistically increase the efficiency of antibiotics through their combination with silver nanoparticles (AgNPs). Combinations of four conventional antibiotics including CAZ, imipenem (IMI), meropenem (MER), and gentamicin sulfate (GENT) with starch-stabilized AgNPs were tested for their antibacterial effects against three isolates of B. pseudomallei. The combination of each antibiotic with AgNPs featured fractional inhibitory concentration (FIC) index values and fractional bactericidal concentration (FBC) index values ranging from 0.312 to 0.75 µg/mL and 0.252 to 0.625 µg/mL, respectively, against the three isolates of B. pseudomallei. The study clearly showed that most of the combinatorial treatments exhibited synergistic antimicrobial effects against all three isolates of B. pseudomallei. The highest enhancing effect was observed for GENT with AgNPs. These results confirmed the combination of each antibiotic with AgNPs restored their bactericidal potency in the bacterial strains that had previously been shown to be resistant to the antibiotics. In addition, morphological changes examined by SEM confirmed that the bacterial cells were severely damaged by combinations at the FBC level. Although bacteria produce fibers to protect themselves, ultimately the bacteria were killed by the antibiotic-AgNPs combinations. Overall, these results suggest the study of antibiotic-AgNPs combinations as an alternative design strategy for potential therapeutics to more effectively combat the melioidosis pathogen.

AFM Study of Nanoscale Membrane Perturbation Induced by Antimicrobial Lipopeptide C14 KYR.

Lipopeptides have been extensively studied as potential antimicrobial agents. In this study, we focused on the C14-KYR lipopeptide, a modified version of the KYR tripeptide with myristic acid at the N-terminus. Here, membrane perturbation of live E. coli treated with the parent KYR and C14-KYR peptides was compared at the nanoscale level using AFM imaging. AFM analyses, including average cellular roughness and force spectroscopy, revealed the severe surface disruption mechanism of C14-KYR. A loss of surface roughness and changes in topographic features included membrane shrinkage, periplasmic membrane separation from the cell wall, and cytosolic leakage. Additional evidence from synchrotron radiation FTIR microspectroscopy (SR-FTIR) revealed a marked structural change in the membrane component after lipopeptide attack. The average roughness of the E. coli cell before and after treatment with C14-KYR was 129.2 ± 51.4 and 223.5 ± 14.1 nm, respectively. The average rupture force of the cell treated with C14-KYR was 0.16 nN, four times higher than that of the untreated cell. Our study demonstrates that the mechanistic effect of the lipopeptide against bacterial cells can be quantified through surface imaging and adhesion force using AFM.

Tannic Acid-Stabilized Silver Nanoparticles Used in Biomedical Application as an Effective Antimelioidosis and Prolonged Efflux Pump Inhibitor against Melioidosis Causative Pathogen.

Burkholderia pseudomallei is the causative pathogen of melioidosis and this bacterium is resistant to several antibiotics. Silver nanoparticles (AgNPs) are an interesting agent to develop to solve this bacterial resistance. Here, we characterize and assess the antimelioidosis activity of AgNPs against these pathogenic bacteria. AgNPs were characterized and displayed a maximum absorption band at 420 nm with a spherical shape, being well-monodispersed and having high stability in solution. The average size of AgNPs is 7.99 ± 1.46 nm. The antibacterial efficacy of AgNPs was evaluated by broth microdilution. The bactericidal effect of AgNPs was further assessed by time-kill kinetics assay. Moreover, the effect of AgNPs on the inhibition of the established biofilm was investigated by the crystal violet method. In parallel, a study of the resistance induction development of B. pseudomallei towards AgNPs with efflux pump inhibiting effect was performed. We first found that AgNPs had strong antibacterial activity against both susceptible and ceftazidime-resistant (CAZ-resistant) strains, as well as being efficiently active against B. pseudomallei CAZ-resistant strains with a fast-killing mode via a bactericidal effect within 30 min. These AgNPs did not only kill planktonic bacteria in broth conditions, but also in established biofilm. Our findings first documented that the resistance development was not induced in B. pseudomallei toward AgNPs in the 30th passage. We found that AgNPs still showed an effective efflux pump inhibiting effect against these bacteria after prolonged exposure to AgNPs at sublethal concentrations. Thus, AgNPs have valuable properties for being a potent antimicrobial agent to solve the antibiotic resistance problem in pathogens.

The Decoration of ZnO Nanoparticles by Gamma Aminobutyric Acid, Curcumin Derivative and Silver Nanoparticles: Synthesis, Characterization and Antibacterial Evaluation.

Zinc oxide nanoparticles (ZnO NPs) are applied in various applications in catalysis, biosensing, imaging, and as antibacterial agents. Here we to prepare ZnO nanomaterials decorated by γ-amino butyric acid (GABA), curcumin derivatives (CurBF2) and silver nanoparticles (CurBF2-AgNPs). The structures of all ZnO nanostructures were characterized using Fourier transform infrared (FT-IR) spectroscopy, X-ray diffraction (XRD), UV-VIS spectrophotometry, fluorescence spectrophotometry, scanning electron microscopy (SEM), transmission electron microscopy (TEM), and high-resolution transmission electron microscopy (HR-TEM). Further, their antibacterial activities against Gram-negative (Escherichia coli) and Gram-positive (Staphylococcus aureus) bacteria were investigated through analysis of minimum inhibitory concentration (MIC) method. Among the prepared nanostructures, the ZnO NPs-GABA/CurBF2-AgNPs showed excellent antibacterial activity against both Gram-positive and -negative bacteria. ZnO NPs fabricated here may have potential use in future anti-bacterial compositions and coatings technologies.

Correlation of age and sex with urine dehydroepiandrosterone sulfate level in healthy Thai volunteers.

OBJECTIVE: Dehydroepiandrosterone sulfate (DHEAs), a prohormone secreted by the adrenal gland, plays a role in the synthesis of sex hormones, namely, androgen and estrogen. It has been found that the amount of DHEAs is correlated with age, although most studies have focused on the correlation of serum DHEAs levels with age and sex. Thus, this noninvasive, cross-sectional study aimed to investigate the correlation of urine DHEAs levels with age and sex in healthy Thai volunteers aged 20-80 years. METHODS: DHEAs levels were measured in 178 healthy volunteers using electrochemiluminescence immunoassay and then normalized by creatinine. Multiple regression was performed to determine the correlation of urine DHEAs levels normalized by creatinine with age and sex. RESULTS: The normalized DHEAs levels are correlated with age group for both sexes. Moreover, an increasing trend in DHEAs levels was found in the age group 20-29 years, and the DHEAs level peaked at the age group 30-39 years before declining with advancing age. Based on the multiple regression analyses, the significance of the interaction term (P â€‹< â€‹0.05) indicates that both age and sex significantly contribute to the prediction of ln (DHEAs/Creatinine). Our fitted model implies the following: as age increases by 1 year, DHEAs/Creatinine is expected to decrease by 3.63% in females and by 2.18% in males. CONCLUSION: This study reports more data on clinical reference value of urine DHEAs levels in healthy volunteers. Our result demonstrates urine DHEAs levels are associated with age and sex and decline by 2-3% a year.

Colorimetric detection of Hg(II) by γ-aminobutyric acid-silver nanoparticles in water and the assessment of antibacterial activities.

In this work, we report the synthesis of silver nanoparticles (AgNPs) via a wet-chemical reduction procedure using citrate (Cit) and γ-aminobutyric acid (GABA) as stabilizers. The formation of GABA-Cit@AgNPs was confirmed by UV-vis spectroscopy with a surface plasmon resonance band at 393 nm clearly confirming the formation of silver nanoparticles. AgNPs were characterized using UV-vis spectroscopy, attenuated total reflectance-fourier transform infrared spectroscopy (ATR-FTIR), transmission electron microscope (TEM), energy-dispersive X-ray spectroscopy (EDX), X-ray diffraction (XRD), dynamic light scattering (DLS), and zeta potential. The as-prepared AgNPs can be used for the detection of hazardous mercury ions (Hg2+) in water by colorimetric method with a limit of detection (LOD) and limit of quantitation (LOQ) of 2.37 µM and 3.99 µM, respectively. The linear working range for Hg2+ detection is 5-35 µM and the sensor probe was applied to investigate Hg2+ in real drinking water samples with satisfied results. Rapid response to Hg2+ is also observed when the nanoparticles are composited within hydrogels. Moreover, GABA-Cit@AgNPs shows antibacterial activity against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus). The fast and sensitive response of the proposed Hg2+ sensor, together with its antibacterial activities, makes GABA-Cit@AgNPs potentially applicable for the development of cheap, portable, colorimetric sensors in fieldwork.

Heterologous expression and mutagenesis of recombinant Vespa affinis hyaluronidase protein (rVesA2).

BACKGROUND: Crude venom of the banded tiger waspVespa affinis contains a variety of enzymes including hyaluronidases, commonly known as spreading factors. METHODS: The cDNA cloning, sequence analysis and structural modelling of V. affinis venom hyaluronidase (VesA2) were herein described. Moreover, heterologous expression and mutagenesis of rVesA2 were performed. RESULTS: V. affinis venom hyaluronidase full sequence is composed of 331 amino acids, with four predicted N-glycosylation sites. It was classified into the glycoside hydrolase family 56. The homology modelling exhibited a central core (α/ß)7 composed of Asp107 and Glu109, acting as the catalytic residues. The recombinant protein was successfully expressed in E. coli with hyaluronidase activity. A recombinant mutant type with the double point mutation, Asp107Asn and Glu109Gln, completely lost this activity. The hyaluronidase from crude venom exhibited activity from pH 2 to 7. The recombinant wild type showed its maximal activity at pH 2 but decreased rapidly to nearly zero at pH 3 and was completely lost at pH 4. CONCLUSION: The recombinant wild-type protein showed its maximal activity at pH 2, more acidic pH than that found in the crude venom. The glycosylation was predicted to be responsible for the pH optimum and thermal stability of the enzymes activity.

Heterologous expression and mutagenesis of recombinant Vespa affinis hyaluronidase protein (rVesA2)

Background:Crude venom of the banded tiger waspVespa affinis contains a variety of enzymes including hyaluronidases, commonly known as spreading factors.Methods:The cDNA cloning, sequence analysis and structural modelling of V. affinis venom hyaluronidase (VesA2) were herein described. Moreover, heterologous expression and mutagenesis of rVesA2 were performed.Results:V. affinis venom hyaluronidase full sequence is composed of 331 amino acids, with four predicted N-glycosylation sites. It was classified into the glycoside hydrolase family 56. The homology modelling exhibited a central core (α/β)7 composed of Asp107 and Glu109, acting as the catalytic residues. The recombinant protein was successfully expressed in E. coli with hyaluronidase activity. A recombinant mutant type with the double point mutation, Asp107Asn and Glu109Gln, completely lost this activity. The hyaluronidase from crude venom exhibited activity from pH 2 to 7. The recombinant wild type showed its maximal activity at pH 2 but decreased rapidly to nearly zero at pH 3 and was completely lost at pH 4.Conclusion:The recombinant wild-type protein showed its maximal activity at pH 2, more acidic pH than that found in the crude venom. The glycosylation was predicted to be responsible for the pH optimum and thermal stability of the enzymes activity.