Solenopsis geminata is recognized for containing the allergenic proteins Sol g 1, 2, 3, and 4 in its venom. Remarkably, Sol g 2.1 exhibits hydrophobic binding and has a high sequence identity ...(83.05%) with Sol i 2 from S. invicta. Notably, Sol g 2.1 acts as a mediator, causing paralysis in crickets. Given its structural resemblance and biological function, Sol g 2.1 may play a key role in transporting hydrophobic potent compounds, which induce paralysis by releasing the compounds through the insect’s nervous system. To investigate this further, we constructed and characterized the recombinant Sol g 2.1 protein (rSol g 2.1), identified with LC-MS/MS. Circular dichroism spectroscopy was performed to reveal the structural features of the rSol g 2.1 protein. Furthermore, after treating crickets with S. geminata venom, immunofluorescence and immunoblotting results revealed that the Sol g 2.1 protein primarily localizes to the neuronal cell membrane of the brain and thoracic ganglia, with distribution areas related to octopaminergic neuron cell patterns. Based on protein—protein interaction predictions, we found that the Sol g 2.1 protein can interact with octopamine receptors (OctRs) in neuronal cell membranes, potentially mediating Sol g 2.1’s localization within cricket central nervous systems. Here, we suggest that Sol g 2.1 may enhance paralysis in crickets by acting as carriers of active molecules and releasing them onto target cells through pH gradients. Future research should explore the binding properties of Sol g 2.1 with ligands, considering its potential as a transporter for active molecules targeting pest nervous systems, offering innovative pest control prospects.
Polydopamine (PDA) has now been widely applied to electrochemical biosensing because of its excellent biocompatibility, abundant functional groups, and facile preparation. In this study, polydopamine ...nanoparticles (PDA-NPs)-functionalized electrochemical aptasensor was developed for the rapid, sensitive, and cost-effective detection of glycated albumin (GA), a promising biomarker for glycemic control in diabetic patients. PDA-NPs were synthesized at various pH conditions in Tris buffer. Cyclic voltammetry (CV) of PDA-NPs-coated screen-printed carbon electrodes (SPCEs) revealed that the materials were more conductive when PDA-NPs were synthesized at pH 9.5 and 10.5 than that at pH 8.5. At pH 10.5, the prepared PDA and PDA-aptamer NPs were monodispersed spherical morphology with an average size of 118.0 ± 1.9 and 127.8 ± 2.0 nm, respectively. When CV and electrochemical impedance spectrometry (EIS) were used for the characterization and detection of the electrochemical aptasensor under optimal conditions, the proposed aptasensor exhibited a broad linearity for detection of GA at a clinically relevant range of (1-10,000 µg mL
), provided a low detection limit of 0.40 µg mL
, appreciable reproducibility (less than 10%), and practicality (recoveries 90-104%). In addition, our developed aptasensor presented a great selectivity towards GA, compared to interfering substances commonly present in human serum, such as human serum albumin, urea, glucose, and bilirubin. Furthermore, the evaluation of the aptasensor performance against GA-spiked serum samples showed its probable applicability for clinical use. The developed PDA aptasensor demonstrated excellent sensitivity and selectivity towards GA detection with a simple and facile fabrication process. This proposed technique shows its potential application in GA measurement for improving the screening and management of diabetic patients in the future.
Staphylococcus aureus, especially methicillin-resistant S. aureus (MRSA), is an important bacterium that causes community and healthcare-related infections throughout the world. However, the current ...conventional detection methods are time-consuming. We therefore developed and evaluated a recombinase polymerase amplification-lateral flow strip (RPA-LF) approach for detection of MRSA in positive blood-culture samples. Sixty positive blood-cultures from a hospital were tested directly without DNA extraction and purification before the amplification reaction. RPA primers and probes were designed for nuc (encoding thermonuclease) and mecA (encoding penicillin-binding protein 2a) genes to diagnose S. aureus and its methicillin-resistance status. The RPA reaction occurred under isothermal conditions (45°C) within 20 min and a result was provided by the LF strip in a further 5 min at room temperature. The evaluation of RPA-LF using blood-culture samples showed 93.3% (14/15) sensitivity for identifying S. aureus, and no cross-amplification was seen 100% (45/45) specificity. For detection of methicillin resistance, the RPA-LF test provided 100% (16/16) sensitivity and 97.7% (43/44) specificity. The RPA-LF is rapid, highly sensitive, robust and easy to use. It can be used for direct detection of MRSA with no requirement for special equipment.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
The increasing antimicrobial-resistant prevalence has become a severe health problem. It has led to the invention of a new antimicrobial agent such as antimicrobial peptides. Heteroscorpine-1 is an ...antimicrobial peptide that has the ability to kill many bacterial strains. It consists of 76 amino acid residues with a cecropin-like region in N-terminal and a defensin-like region in the C-terminal. The cecropin-like region from heteroscorpine-1 (CeHS-1) is similar to cecropin B, but it lost its glycine-proline hinge region. The bioinformatics prediction was used to help the designing of mutant peptides. The addition of glycine-proline hinge and positively charged amino acids, the deletion of negatively charged amino acids, and the optimization of the hydrophobicity of the peptide resulted in two mutant peptides, namely, CeHS-1 GP and CeHS-1 GPK. The new mutant peptide showed higher antimicrobial activity than the native peptide without increasing toxicity. The interaction of the peptides with the membrane showed that the peptides were capable of disrupting both the inner and outer bacterial cell membrane. Furthermore, the SEM analysis showed that the peptides created the pore in the bacterial cell membrane resulted in cell membrane disruption. In conclusion, the mutants of CeHS-1 had the potential to develop as novel antimicrobial peptides.
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.
Phospholipase A1 from Thai banded tiger wasp (Vespa affinis) venom also known as Ves a 1 plays an essential role in fatal vespid allergy. Ves a 1 becomes an important therapeutic target for toxin ...remedy. However, established Ves a 1 structure or a mechanism of Ves a 1 function were not well documented. This circumstance has prevented efficient design of a potential phospholipase A1 inhibitor. In our study, we successfully recruited homology modeling and molecular dynamic (MD) simulation to model Ves a 1 three-dimensional structure. The Ves a 1 structure along with dynamic behaviors were visualized and explained. In addition, we performed molecular docking of Ves a 1 with 1,2-Dimyristoyl-sn-glycero-3-phosphorylcholine (DMPC) lipid to assess a possible lipid binding site. Interestingly, molecular docking predicted another lipid binding region apart from its corresponding catalytic site, suggesting an auxiliary role of the alternative site at the Ves a 1 surface. The new molecular mechanism related to the surface lipid binding site (auxiliary site) provided better understanding of how phospholipase A1 structure facilitates its enzymatic function. This auxiliary site, conserved among Hymenoptera species as well as some mammalian lipases, could be a guide for interaction-based design of a novel phospholipase A1 inhibitor.
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•Edible bird’s nest (EBN) specifically promotes the expansion of T-cells both in vitro and in vivo studies.•EBN treatments can maintain the homeostasis of T-cells under the ...immunosuppressive condition.•The level of serum IL-2 has predominantly increased in response to the administration of EBN treatments.•The changes of secondary lymphoid organs (Peyer’s patches and spleen) have been affected by EBN treatments.
This study aimed to investigate the immunomodulatory effect of edible bird’s nest (EBN) in vitro and in vivo studies. EBN-treated peripheral blood mononuclear cells showed that EBN specifically enhanced the expansion of CD3+ T-cells. The restoration of lymphocyte subpopulations under the influence of immunosuppressive drug has been successfully recovered in CD3+ T-cells, not CD45RA+ B-cells and CD335+ NK-cells. In addition, oral administration of EBNs in Sprague-Dawley rats revealed their potential to increase the number of peripheral blood T-cells. Our study also demonstrated that EBN treatments affected the numbers of the Peyer’s patches, spleen weight and length, and cellularity of the periarteriolar lymphoid sheath. Interestingly, we observed that elevation of serum interleukin-2 (IL-2) had been correlated with the proliferation of T-cells in the animal model. Therefore, these results are essential for developing therapeutic strategies in improving immunity, particularly T-cell homeostasis, under immunosuppressive therapy.
Chitosan or polyvinyl pyrrolidone (PVP) were used in combination with hydroxypropyl methylcellulose (HPMC) and poloxamer 407 (P407) as gelling agents for oral drug delivery. The performance ...interaction with mucin of chitosan-composed gel (F1) and PVP-composed gel (F2) was compared using attenuated total reflectance-Fourier-transform infrared (ATR-FTIR) spectroscopy at controlled temperatures of 25 and 37 °C for 1 and 5 min. F1 containing niosome-entrapped melatonin or its derivatives was investigated for mucoadhesive interaction on mucosa by ATR-FTIR spectroscopy under the same conditions. The results showed that F1-treated mucin gave a significantly lower amide I/amide II ratio than untreated mucin and F2-treated mucin did within 1 min, suggesting improved rapid affinity between mucin and chitosan. The spectra of mucosa treated with F1 incorporating niosomes of melatonin or its derivatives showed peak shifts at C=O (amide I), N-H (amide II), and carbohydrate regions and an associated decrease in the amide I/amide II ratio and increase in the carbohydrate/amide II ratio. These results indicated electrostatic interaction and hydrogen bonding between chitosan and mucin on the mucosa. In conclusion, the molecular interaction between gels and mucin/mucosa detected at amide I and amide II of proteins and the carbohydrate region could lead to an improved mucoadhesive property of the gel on the mucosa.
Known antimicrobial peptides KT2 and RT2 as well as the novel RP9 derived from the leukocyte extract of the freshwater crocodile (
Crocodylus siamensis
) were used to evaluate the ability in killing ...human cervical cancer cells. RP9 in the extract was purified by a combination of anion exchange column and reversed-phase HPLC, and its sequence was analyzed by mass spectrometry. The novel peptide could inhibit Gram-negative
Vibrio cholerae
(clinical isolation) and Gram-positive
Bacillus pumilus
TISTR 905, and its MIC values were 61.2 µM. From scanning electron microscopy, the peptide was seen to affect bacterial surfaces directly. KT2 and RT2, which are designed antimicrobial peptides using the
C. siamensis
Leucrocin I template, as well as RP9 were chemically synthesized for investigation of anticancer activity. By Sulforhodamine B colorimetric assay, these antimicrobial peptides could inhibit both HeLa and CaSki cancer cell lines. The IC
50
values of KT2 and RT2 for HeLa and CaSki cells showed 28.7–53.4 and 17.3–30.8 µM, while those of RP9 were 126.2 and 168.3 µM, respectively. Additionally, the best candidate peptides KT2 and RT2 were used to determine the apoptotic induction on cancer cells by human apoptosis array assay. As a result, KT2 and RT2 were observed to induce apoptotic cell death in HeLa cells. Therefore, these results indicate that KT2 and RT2 with antimicrobial activity have a highly potent ability to kill human cervical cancer cells.
is a significant cause of urinary tract infection that may contribute to struvite stones. Anti-infection of this bacterium and anti-struvite formation must be considered.
Burm. F. (SA) has been used ...for the treatment of diseases related to kidneys. Therefore, we investigated the effects of the SA leaf ethanolic extract (SAEE) on growth and on virulent factors (swarming motility and urease activity) of
isolated from kidney stone formers. We also evaluated anti-struvite crystal formation and phytochemical constituents of SAEE. The minimum inhibitory concentrations (MICs) of SAEE against three clinical
isolates were 8 mg/mL. Intriguingly, the 1/2MIC of SAEE had significant inhibitory effects on the swarming motility and urease activity of clinical
isolates when compared with the condition without SAEE. The SAEE at the various concentrations significantly inhibited the average weights of struvite crystals in a dose-dependent manner, compared with the control. The phytochemical analysis revealed that SAEE contained catechin, chlorogenic acid, rutin, and ferulic acid. This study indicated that SAEE has anti-
and anti-struvite crystal activities via its bioactive compounds. For this reason, SAEE may be developed as a new agent for the treatment of struvite stone induced by
.
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