Potential Candidate Research Articles

Genipin-Cross-Linked Silk Fibroin/Alginate Dialdehyde Hydrogel with Tunable Gelation Kinetics, Degradability, and Mechanical Properties: A Potential Candidate for Tissue Regeneration.

Genipin-cross-linked silk fibroin (SF) hydrogel is considered to be biocompatible and mechanically robust. However, its use remains a challenge for in situ forming applications due to its prolonged gelation process. In our attempt to facilitate the in situ fabrication of a genipin-mediated SF hydrogel, alginate dialdehyde (ADA) was utilized as a reinforcement template. Here, SF/ADA-based hydrogels with different compositions were synthesized covalently and ionically. Incorporating ADA into the SF hydrogel increased pore size (44.66-174.66 μm), porosity (61.59-80.40%), and the equilibrium swelling degree (7.60-30.17). Moreover, a wide range of storage modulus and compressive modulus were obtained by adjusting the proportions of SF and ADA networks within the hydrogel. The in vitro cell analysis using preosteoblast cells (MC3T3-E1) demonstrated the cytocompatibility of all hydrogels. Overall, the covalently and ionically cross-linked SF/ADA hydrogel represents a promising solution for in situ forming hydrogels for applications in tissue regeneration.

(032) Prevalence of Testosterone Deficiency and Risk Factors in Patients with Localized Prostate Cancer After Radical Prostatectomy

Abstract Introduction Testosterone therapy (TTh) is contra-indicated in patients with prostate cancer (PCa), however according to the recent AUA guideline, TTh can be offer for localized PCa with no evidence of active disease. However, it is not known how much localized PCa patients suffer from testosterone (T) deficiency after treatment in real clinical practice, and further clinical characteristics on those potential candidates for TTh is insufficient. Objective This study analyzed T profile of patients with localized PCa who underwent radical prostatectomy (RP) to determined the prevalence of testosterone deficiency and its risk factors. Methods We reviewed patients who underwent RP for localized PCa between 2016 and 2021. Patients with less than 1 year of follow-up and patients with biochemical recurrence or distant metastasis during follow-up were excluded. In addition, we exclude the patient who had no T profile preoperatively and postoperative 1 year. The serum T level threshold for T deficiency was defined as 3.0 ng/mL. The primary outcome was the prevalence of TDS, and the secondary outcome was the predicting factors of TDS. Results A total of 192 patients underwent RP with localized PCa during the study period. Among them, 120 patients were analyzed, excluding patients without baseline T and follow-up T level at postoperative 1 year (n=67) and patients with BCR during the follow-up (n=5). Eighteen of 120 patients (15.0%) had T deficiency before RP. Excluding preoperative T deficiency, 33 (32.4%) of 102 patients had their T level reduced to less than 3.0 ng/ml after RP. Among them, 16 patients (15.7%) had persistent decreased T level. Table 1. Summarized the baseline characteristics of patients with and without T deficiency after RP. BMI (OR: 1.586, p< 0.001) was statistically significant in multivariate logistic regression to identify the predicting factor for T deficiency after RP. Conclusions In the patients with localized prostate cancer, there were a significant number of TTh candidates. In particular, obese patients had a higher risk of T deficiency. Disclosure No.

Genome-wide association study revealed the reason for the decrease in grain iron concentration during wheat breeding process in China

Iron (Fe) deficiency affects over two billion people worldwide. Developing high-yielding wheat (Triticum aestivum L.) cultivars with high grain Fe concentration (GFeC) is a key step toward alleviating malnutrition in humans. In this study, we used a diversity panel of 388 wheat accessions to examine the GFeC in four field environments. We found that the mean GFeC of the four environments decreased with the release year. And GFeC showed poor correlation with grain yield or the three yield factors in the four environments. Importantly, wheat cultivars in the major wheat-producing region of China showed the lowest GFeC. The GFeC phenotype was used for a genome-wide association study (GWAS) using a 660 K single-nucleotide polymorphism (SNP) chip, and 17 quantitative trait loci (QTLs) were identified on chromosomes 3B, 4 A, 5 A, 6B, 6D, 7 A, 7B, and 7D. However, no universal QTLs were detected among the 17 identified QTLs and 2280 yield-related QTLs. Through allelic analysis, two homologous genes, TraesCS7A01G197400 and TraesCS7D01G198900, were identified as potential candidate genes for GFeC, while the favourable alleles for GFeC were lost during the selection breeding process in China. The insights gained from this study showed that the yield dilution effect was not applied to the decrease in GFeC, which should be mainly caused by the loss of favourable alleles during the wheat breeding process in China. Therefore, wheat breeders should pay more attention to other traits to improve GFeC rather than the yield dilution effect.

Space-confined intercalation expansion strategy for simple and rapid synthesis of kish-based expanded graphite for aluminum ion batteries

The practical application of graphite cathodes in aluminum ion batteries (AIBs) is hindered by poor cycle and rate performance of graphite, due to serious structural degradation. In this work, different types of kish-expanded graphite (KEG) with different expansion volumes were prepared simply and rapidly using low-cost solid waste kish graphite (KG) as precursor, and K2S2O8 and concentrated H2SO4 as oxide intercalation agents by space-confined intercalation expansion strategy. By adjusting the mass ratio of K2S2O8 to KG, the macrostructure and microstructure of KEG can be effectively controlled. Specifically, KEG6, synthesized with the mass ratio of K2S2O8 to KG of 6, exhibited a large expansion volume, graphene nanosheets with numerous graphite microcrystalline layer structures, and micro-nano interlayer pore structures resulting from the cross-linking of graphene nanosheets, along with abundant oxygen-containing functional groups. KEG6, as a cathode material for AIBs, exhibited outstanding electrochemical performance, including superior specific capacity (141.1 mA h/g at 50 mA/g), good rate capability (119.8 mA h/g at 1000 mA/g, 85.6 mA h/g at 2000 mA/g) and remarkable cycle stability (134.0 mA h/g with 99.1% Coulombic efficiency after 10,000 cycles at 1000 mA/g). This study provides a simple and rapid method for the large-scale synthesis of KEG as a potential candidate for AIBs.

Matairesinoside, a novel inhibitor of TMEM16A ion channel, loaded with functional hydrogel for lung cancer treatment

The incidence and mortality rates of lung cancer have remained high for several decades, necessitating the discovery of new drugs and the development of effective treatment strategies. This study identified matairesinoside (MTS) as a potent inhibitor of TMEM16A, a novel drug target for lung cancer. Molecular simulation combined with site-directed mutagenesis experiments confirmed the key binding sites of MTS and TMEM16A. Cell experiments demonstrated that MTS significantly inhibited the growth, migration, and invasion of lung cancer cells, while inducing apoptosis. Gene knockdown and overexpression studies further revealed that TMEM16A is the target for MTS in regulating lung cancer cell growth. Western blot analysis elucidated the signaling transduction network involved in MTS-mediated regulation of lung cancer. Building upon these findings, a biodegradable self-healing functional hydrogel was developed to load MTS, aiming to enhance therapeutic efficacy and minimize side effects in vivo. Animal experiments demonstrated that the hydrogel/MTS formulation exhibited satisfactory inhibitory effects on lung cancer and mitigated the side effects associated with direct MTS injection. This study identified MTS as a potential candidate for anti-lung cancer therapy with well-defined pharmacological mechanisms. Moreover, the targeted drug delivery system utilizing the hydrogel/MTS platform offers a promising approach for lung cancer treatment.

Preparation of Sea Urchin‐Like Co/Ni Alloy Compounds for Pseudocapacitive Supercapacitor

Energy density and power density are the two most crucial metrics in advanced electrochemical energy storage (EES) devices. The design of pseudocapacitor materials with good structure and reasonable composition can ensure that the power density can be greatly increased under the condition of little or no reduction in energy density. However, how to reasonably design excellent pseudocapacitor materials is still a huge challenge. Herein, layered double hydroxides containing Ni and Co elements on carbon materials derived from distillers’ grains by hydrothermal synthesis (NiCo‐LDH/JKPC‐2) are grown. The fabricated NiCo‐LDH/JKPC‐2 has exhibited a sea urchin‐like nanostructure that primarily consists of mesoporous and possesses a high specific surface area (303 m2 g−1), which facilitates the formation of larger active sites, thereby increasing the rate of charge transport. The specific capacitance of the NiCo‐LDH/JKPC‐2 reaches a maximum value of 1454 F g−1 at 1 A g−1. When it combines with carbon materials originating from distillers’ grains, the assembled asymmetric supercapacitor electrode exhibits a remarkable energy density (40.86 Wh kg−1 at 775 W kg−1) and superior power density (7750.8 W kg−1 at 21.53 Wh kg−1). Thus, the NiCo‐LDH/JKPC‐2 asymmetric supercapacitor electrode can be considered as a potential candidate in advanced energy storage device systems.

Inhibition of PRMT1 Suppresses the Growth of U87MG-Derived Glioblastoma Stem Cells by Blocking the STAT3 Signaling Pathway.

Glioblastoma stem cells (GSCs) play a pivotal role in the initiation, progression, resistance to treatment, and relapse of glioblastoma multiforme (GBM). Thus, identifying potential therapeutic targets and drugs that interfere with the growth of GSCs may contribute to improved treatment outcomes for GBM. In this study, we first demonstrated the functional role of protein arginine methyltransferase 1 (PRMT1) in GSC growth. Furamidine, a PRMT1 inhibitor, effectively inhibited the proliferation and tumorsphere formation of U87MG-derived GSCs by inducing cell cycle arrest at the G0/G1 phase and promoting the intrinsic apoptotic pathway. Moreover, furamidine potently suppressed the in vivo tumor growth of U87MG GSCs in a chick embryo chorioallantoic membrane model. In particular, the inhibitory effect of furamidine on U87MG GSC growth was associated with the downregulation of signal transducer and activator of transcription 3 (STAT3) and key GSC markers, including CD133, Sox2, Oct4, Nanog, aldehyde dehydrogenase 1, and integrin α6. Our results also showed that the knockdown of PRMT1 by small interfering RNA significantly inhibited the proliferation of U87MG GSCs in vitro and in vivo through a molecular mechanism similar to furamidine. In addition, combined treatment with furamidine and berbamine, a calcium/calmodulin-dependent protein kinase II gamma (CaMKIIγ) inhibitor, inhibited the growth of U87MG GSCs more strongly than single-compound treatment. The increased antiproliferative effect of combining the two compounds resulted from a stronger downregulation of STAT3-mediated downstream GBM stemness regulators through dual PRMT1 and CaMKIIγ function blockade. In conclusion, these findings suggest that PRMT1 and its inhibitor, furamidine, are potential novel therapeutic targets and drug candidates for effectively suppressing GSC growth.

Unveiling the influence of various vacancies on the mechanical properties of Ti4C3O2

Two-dimensional transition metal carbides and nitrides are widely recognized as potential candidates in areas such as energy storage and sensors. In this work, we explore the mechanical properties of Ti4C3O2 with a perfect cell and Ti4C3O2 with vacancies by a first-principles method. The results indicate that the loss of C in the outer layer of Ti4C3O2 is more likely to occur due to the smaller formation energy of C-vacancy. By considering the charge contribution, the removed atom changes the valence electron equilibrium concentration between that atom and its neighbors, which in turn affects the mechanical properties of Ti4C3O2. The presence of various vacancies affects the strength of the TiO bonds on the shear plane, causing Ti4C3O2 to exhibit changes towards brittleness or ductility. It is noteworthy that the elastic modulus, critical strain, and ideal stress of Ti4C3O2 with different concentrations of O vacancies are all approximately the same, suggesting that the variations in their mechanical properties are not significant for Ti4C3O2 with different degrees of O functional group coverage. Our findings offer valuable insights for the development of MXene-based nanoflexible energy storage devices, gas-sensitive sensors, high-temperature environmental applications, and beyond.

Mechanism exploration of Zoledronic acid combined with PD-1 in the treatment of hepatocellular carcinoma

How to increase the response of immune checkpoint inhibitors (ICIs) is a challenge. In clinical, we found that Zoledronic acid (ZA) may increase the anti-tumor effect of immunotherapy for hepatocellular carcinoma (HCC). To explore the underlying mechanism, we established a mouse model of HCC by subcutaneously injecting Hepa1-6 cell line. The result showed that the tumor volume in the ZA plus anti-PD-1 monocloning antibody (anti-PD-1 mAb) treatment groups was significantly smaller than that of control group, and the onset time of tumor inhibition was even shorter than that of the anti-PD-1 mAb group. Using flow cytometry (FC) to detect the proportion of major immune cell subsets in tumor tissues of each group of mice, we found that the synergistic anti-tumor effect of ZA and anti-PD-1 mAb may be related to ZA-induced polarization of macrophages toward the M1 phenotype. Next, we performed bulk RNA sequencing on tumor samples from different groups to obtain differentially expressed genes (DEGs), which were then input DEGs into pathway enrichment analysis. Data indicated that ZA participated in the M1-type polarization via ferroptosis-related pathways. Our results revealed how ZA involves in the anti-tumor effect of PD-1 monoclonal antibody and provided a potential therapeutic candidate for patients with HCC.

Development of activation-tagged gain-of-functional mutants in indica rice line (BPT 5204) for sheath blight resistance.

The development of sheath blight (ShB) resistance varieties has been a challenge for scientists for long time in rice. Activation tagging is an efficient gain-of-function mutation approach to create novel phenotypes and to identify their underlying genes. In this study, a mutant population was developed employing activation tagging in the recalcitrant indica rice (Oryza sativa L.) cv. BPT 5204 (Samba Mahsuri) through activation tagging. In this study, we have generated more than 1000 activation tagged lines in indica rice, from these mutant population 38 (GFP- RFP+) stable Ds plants were generated through germinal transposition at T2 generation based on molecular analysis and seeds selected on hygromycin (50mg/L) containing medium segregation analyses confirmed that the transgene inherited as mendelian segregation ratio of 3:1 (3 resistant: 1 susceptible). Of them, five stable activation tagged Ds lines (M-Ds-1, M-Ds-2, M-Ds-3, M-Ds-4 and M-Ds-5) were selected based on phenotypic observation through screening for sheath blight (ShB) resistance caused by fungal pathogen Rhizoctonia solani (R. solani),. Among them, M-Ds-3 and M-Ds-5 lines showed significant resistance for ShB over other tagged lines and wild type (WT) plants. Furthermore, analysed for launch pad insertion through TAIL-PCR results and mapped on corresponding rice chromosomes. Flanking sequence and gene expression analysis revealed that the upregulation of glycoside hydrolase-OsGH or similar to Class III chitinase homologue (LOC_Os08g40680) in M-Ds-3 and a hypothetical protein gene (LOC_Os01g55000) in M-Ds-5 are potential candidate genes for sheath blight resistance in rice. In the present study, we developed Ac-Ds based ShB resistance gain-of-functional mutants through activation tagging in rice. These activation tagged mutant lines can be excellent sources for the development of ShB resistant cultivars in rice.

Crosslinked Polyesters as Fully Biobased Coatings with Cutin Monomer from Tomato Peel Wastes.

Cutin, one of the main structural components of tomato peels, is a waxy biopolymer rich in hydroxylated fatty acids. In this study, 10,16-dihydroxyhexadecanoic acid (10,16-diHHDA) was extracted and isolated from tomato peels and exploited to develop fully crosslinked polyesters as potential candidates for replacing fossil-based metal protective coatings. A preliminary screening was conducted to select the base formulation, and then a design of experiments (DoE) was used as a methodology to identify the optimal composition to develop a suitable coating material. Different formulations containing 10,16-diHHDA and other biorefinery monomers, including 2,5-furandicarboxylic acid, were considered. To this end, all polyesters were characterized through differential scanning calorimetry (DSC) and gel content measurements to determine their Tg value and crosslinking efficiency. Compositions exhibiting the best trade-off between Tg value, chemical resistance, and sufficiently high 10,16-diHHDA content between 39 and 48 wt.% were used to prepare model coatings that were characterized for assessing their wettability, scratch hardness, chemical resistance, and adhesion to metal substrates. These polyester coatings showed a Tg in the range of 45-55 °C, a hydrophobic behavior with a water contact angle of around 100°, a good solvent resistance (>100 MEK double rubs), and an adhesion strength to steel higher than 2 MPa. The results obtained confirmed the potential of cutin-based resins as coatings for metal protection, meeting the requirements for ensuring physicochemical properties of the final product, as well as for optimizing the valorization of such an abundant agri-food waste as tomato peels.

Antidiabetic potency and molecular insights of natural products bearing indole moiety: A systematic bioinformatics investigation targeting AKT1

Diabetic mellitus (DM) is a chronic disorder, and type 2 DM (T2DM) is the most prevalent among all categories (nearly 90%) across the globe every year. With the availability of potential drugs, the prevalence rate has remained uncontrollable, while natural resources showed a promising potency, and exploring such potential candidates at the preclinical stage is essential. An extensive literature search selected 89 marine and plant-derived indole derivatives with anti-inflammatory, antioxidant, lipid-lowering, etc., activities. However, as we know, drugs have not been able to convert from ’lead’ to 'mainstream' due to inadequate drug-ability profiles, as our systematic investigation proved and selected herdmanine_A and penerpene_D as the most potential antidiabetic candidates from the library of indole derivatives. Based on our previous network pharmacology study, we selected three new target enzymes: Acetyl-CoA carboxylase 2 (ACACB; PDB ID: 3JRX), cyclin-dependent kinase 4 (CDK4; PDB ID: 3G33), and alpha serine/threonine-protein kinase 1 (AKT1; PDB ID: 3O96) to assess the antidiabetic potency of selected indole derivatives through binding energy or docking score. To conduct molecular docking studies with these enzymes, we used the PyRx-AutoDock platform. Furthermore, molecular dynamic simulation at 100ns, physicochemical analysis, pharmacokinetics, toxicity assessment, and drug-likeness evaluation suggested that herdmanine_A and penerpene_D were the most potent inhibitors against AKT1 compared to koenimbine (most potential based on the recorded IC50 value) and murrayakonine_A (most potential based on the docking score). In summary, herdmanine_A and/or penerpene D have the potential to be used as alternative therapeutic agent for the treatment of diabetes after some pharmacological investigation.

Synthesis of Piperidine Conjugated Quinoxalines as Potential Antibiofilm Agents

Background: The most common cause of food-borne illness is bacterial or viral contamination. Although there are several therapeutics available to combat these microbes, they lost their efficacy in long-term medication. Because, over a period of time, microbes developed resistance against drugs and this antimicrobial resistance is a serious threat to global public health as a consequence of the widely disseminated and careless use of antimicrobials. Therefore, there is a need to develop some new chemical moieties with a safety factor and better efficacy. A series of substituted N-(1-benzylpiperidin-4- yl)quinoxalin-2-amines (5a-j) (5ab, 5ac) were synthesized and screened for their in vitro antibacterial activity against Salmonella paratyphi, a well-known food-borne pathogen. Methods: Experimental methods, agar diffusion and broth microdilution assays were carried out to evaluate the antibacterial activity of the lead compounds. Further, antibiofilm methods, crystal violet, and MTT assays were subjected to investigate their biofilm inhibition capacity against S. paratyphi. Results: Among the tested compounds, 5b, 5e, 5h, and 5j bearing 4-chloro, 3,4-dimethoxy, 4-methyl and thienyl groups on the phenyl ring of quinoxalines emerged as potential candidates having significant antisalmonella activity. In these four potential candidates, compounds 5b and 5h were effective against Salmonella whereas compounds 5e and 5j effectively inhibited the biofilm formation of Salmonella. Conclusion: N-(1-benzylpiperidin-4-yl)quinoxalin-2-amines (5a-j) (5ab, 5ac) were synthesized and evaluated for antisalmonella activity against S. paratyphi. Among the series of compounds, four compounds significantly showed good activity and emerged as antibacterial agents for further studies in the future.

Investigation of work function and quantum capacitance of OH-terminated MXenes as supercapacitor electrodes: A theoretical perspective

MXene has shown promise in supercapacitor electrodes for energy storage devices. As key parameters to develop new electronic devices, the quantum capacitance and work function (WF) of OH-terminated MXenes with 3d, 4d, d transitional metals are investigated by density functional theory. The most stable configurations of 13 M2C(OH)2 MXenes are found, which are further confirmed by negative formation energies. The low WFs of M2C(OH)2 make them become potential candidates as thermionic electron emitters, especially for Sc2C(OH)2 and Lu2C(OH)2 with extra low WFs. Sc2C(OH)2, Cr2C(OH)2, Y2C(OH)2, and Lu2C(OH)2 are considered as negative electrodes, Other nine systems are potential positive electrodes for larger |Qp|/|Qn| than 1, especially for V2C(OH)2, Nb2C(OH)2, and Mo2C(OH)2. Th wide voltage maintains the electrode types of all systems unchanged, but drastically increases the |Qp|/|Qn| of V2C(OH)2. The quantum capacitance of M2CT2 with different terminations is further explored.

A Cytotoxicity Assay as an Alternative to the Murine Model for the Potency Testing of Bothrops jararaca Venom and Antivenom: An Intralaboratory Pre-validation Study.

Antivenom therapy is the only specific treatment for snakebite envenomation, and antivenom potency determination is key in the efficacy assurance quality control process. Nowadays, this process relies on the in vivo murine model - thus, the development of alternative in vitro methods is imperative. In the current study, the principle of the proposed method is the ability of Bothrops venom to induce cytotoxic effects in Vero cells, and the capacity to evaluate the inhibition of this cytotoxicity by the respective antivenom. After exposure to the venom/antivenom, the relative proportions of adherent (viable) cells were evaluated by direct staining with Coomassie Blue. The optical density (OD) of the lysed cell eluate was directly proportional to the number of adherent cells. This cytotoxicity-based alternative method could represent a potential candidate for validation as a replacement for the current in vivo test. The in vitro-determined cytotoxicity of the Brazilian Bothrops reference venom (expressed as the 50% effective concentration; EC50) was 3.61 μg/ml; the in vitro-determined 50% inhibitory concentration (IC50) of the Brazilian Bothrops reference antivenom was 0.133 μl/ml. From these two values, it was possible to calculate the potency of the reference antivenom. The results from the assays exhibited a good linear response, indicating that the method could be a potential candidate replacement method for use in antivenom quality control prior to lot release, subject to further validation.

Computational Repurposing of Potential Dimerization Inhibitors against SARS-CoV-2 Main Protease

Background: The screening, design, and synthesis of various dimerization inhibitors have been an active area of interest for structure-based drug design efforts. Functionally important dimers, such as human immunodeficiency virus (HIV) protease and surviving, are being targeted for such studies over time. Computational repurposing of potential drug candidates provides a cost and time-efficient way in the drug discovery life cycle. Objective: Concerning the current coronavirus disease (COVID-19) scenario, the functionally active dimer of SARS-CoV-2 (severe acute respiratory syndrome) main protease (Mpro) is used as a target to screen possible dimerization inhibitors. Methods: A database of small molecule protein-protein interaction inhibitors was screened for the study. This study used molecular docking, followed by molecular dynamics (MD) simulation and postsimulation binding energy predictions. Results: From the selected 183 compounds, a diazene-based compound and a salicylic-type compound were identified as possible dimerization inhibitors in this study. These two compounds formed stable complexes with the Mpro during the MD simulations. The complexes formed by these two compounds were also unable to form important salt bridge interactions required for the dimerization of the protomers. Conclusion: Experimental studies on both compounds were previously conducted as dimerization inhibitors in HIV. The data led to the possibility of exploring the identified compounds as dimerization inhibitors, which could be important for SARS-CoV-2 therapeutics.

Ivermectin Inhibits Bladder Cancer Cell Growth and Induces Oxidative Stress and DNA Damage.

Bladder cancer is the most common malignant tumor of the urinary system. Nevertheless, current therapies do not provide satisfactory results. It is imperative that novel strategies should be developed for treating bladder cancer. To evaluate the effect of a broad-spectrum anti-parasitic agent, Ivermectin, on bladder cancer cells in vitro and in vivo. CCK-8 and EdU incorporation assays were used to evaluate cell proliferation. Apoptosis was detected by flow cytometry, TUNEL assay, and western blotting. Flow cytometry and DCFH-DA assay were used to analyze the reactive oxygen species (ROS) levels. DNA damage was determined by Neutral COMET assay and γ H2AX expression. Proteins related to apoptosis and DNA damage pathways were determined by WB assay. Xenograft tumor models in nude mice were used to investigate the anti-cancer effect of Ivermectin in vivo. Our study showed that in vitro and in vivo, Ivermectin inhibited the growth of bladder cancer cells. In addition, Ivermectin could induce apoptosis, ROS production, DNA damage, and activate ATM/P53 pathwayrelated proteins in bladder cancer cells. According to these findings, Ivermectin may be a potential therapeutic candidate against bladder cancer due to its significant anti-cancer effect.

Generation of IgG antibodies against Strongyloides stercoralis in mice via immunization with recombinant antigens A133 and Ss-IR.

Strongyloidiasis, caused by the nematode Strongyloides stercoralis, remains a threat to global public health, and a vaccine would be useful to control the disease, especially in developing countries. This study aimed to evaluate the efficacy of recombinant proteins, A133 and Ss-IR, as potential vaccine candidates against strongyloidiasis by investigating the humoral and cellular immune responses in immunized mice. Respective antigens were adjuvanted with Complete Freund's Adjuvant (prime) and Incomplete Freund's Adjuvant (boost) and administered intraperitoneally (prime) and subcutaneously (boost) to female BALB/c mice. For antigen-only doses, only antigens were injected without adjuvants. Altogether, 1 prime dose, 4 booster doses, and 2 antigen-only doses were administered successively. ELISAs were conducted to assess the antibody responses, along with flow cytometry and cytokine ELISA to elucidate the cellular immune responses. Results showed that A133 and Ss-IR induced the production of IgG1 and IgG2a, with A133 generating more robust IgG2a responses than Ss-IR. Flow cytometry findings indicated that effector CD8+T-cells and memory B-cells activity were upregulated significantly for A133 only, whereas cytokine ELISA demonstrated that a Th1/Th2/Th17 mixed cell responses were triggered upon vaccination with either antigen. This preliminary study illustrated the good potential of recombinant A133 and Ss-IR as vaccine candidates against S. stercoralis. It provided information on the probable immune mechanism involved in host defence and the elicitation of protection against S. stercoralis.

Discovery of new N-Phenylamide Isoxazoline derivatives with high insecticidal activity and reduced honeybee toxicity

Isoxazoline is a novel structure with strong potential for controlling agricultural insect pests, but its high toxicity to honeybees limits its development in agriculture. Herein, a series of N-phenylamide isoxazoline derivatives with low honeybee toxicity were designed and synthesized using the intermediate derivatization method. Bioassay results showed that these compounds exhibited good insecticidal activity. Compounds 3b and 3f showed significant insecticidal effects against Plutella xylostella (P. xylostella) with median lethal concentrations (LC50) of 0.06 and 0.07 mg/L, respectively, comparable to that of fluralaner (LC50 = 0.02 mg/L) and exceeding that of commercial insecticide fluxametamide (LC50 = 0.52 mg/L). It is noteworthy that the acute honeybee toxicities of compounds 3b and 3f (LD50 = 1.43 and 1.63 μg/adult, respectively) were significantly reduced to 1/10 of that of fluralaner (LD50 = 0.14 μg/adult), and were adequate or lower than that of fluxametamide (LD50 = 1.14 μg/adult). Theoretical simulation using molecular docking indicates that compound 3b has similar binding modes with fluralaner and a similar optimal docking pose with fluxametamide when binding to the GABA receptor, which may contribute to its potent insecticidal activity and relatively low toxicity to honey bees. This study provides compounds 3b and 3f as potential new insecticide candidates and provides insights into the development of new isoxazoline insecticides exhibiting both high efficacy and environmental safety.

Molecular Docking, In silico ADMET Study and Synthesis of Quinoline Derivatives as Dihydrofolate Reductase (DHFR) Inhibitors: A Solvent-free One-pot Green Approach Through Sonochemistry

Background: Quinoline derivatives have evinced their biological importance in targeting bacteria by inhibiting Dihydrofolate reductase. H2SO4 was successfully applied as an acid catalyst for a green, efficient, and one-pot solvent-free synthesis of quinoline derivatives using sonochemistry approach from various aromatic amines and glycerol with affording yield up to 96% within 6-10 min. Objective: In this study, the synthesis, characterization, and biological assessment of fifteen quinoline derivatives (1-15) as potential DHFR inhibitors were carried out. The target compounds were docked to study the molecular interactions and binding affinities with the 1DLS enzyme. Methods: The synthesized molecules were characterized using IR, MASS, and 1H and 13C NMR. The Insilico molecular docking study was carried out through target Human Dihydrofolate Reductase (DHFR) retrieved from a protein data bank having PDB ID: 1DLS and the antimicrobial activity of all synthesized compounds were tested against Human Dihydrofolate Reductase(DHFR) enzyme by using in-vitro DHFR assay kit. Results: The molecular docking results revealed that compounds 2 and 6 have the lowest binding energy and good binding affinity with the DHFR enzyme. In-silico ADMET predictions revealed that all bestscored compounds had good absorption and drug-like properties for potential use as DHFR inhibitors to treat bacterial infection. The in vitro studies revealed that compounds 2 and 6 show potent DFHR inhibitory activity against gram-positive and gram-negative with IC50 = 12.05 ± 1.55 μM and 10.04 ± 0.73 μM, respectively. While compounds 12, 13, and 15 exhibited moderate antimicrobial activity through DHFR inhibition with IC50= 16.33 ± 0.73 μM, 17.02 ± 1.55 μM, and 18.04 ± 1.05 μM, respectively. Conclusion: This environmentally benign sonochemistry-based approach for synthesizing quinoline derivatives could be affordable for large-scale production and become a potential lead candidate for developing a new quinoline-based antimicrobial agent.