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15 pages, 2365 KiB  
Article
Development of a Novel Aptamer-Antibody Sandwich Chemiluminescent Biosensor and Its Application in the Detection of Aflatoxin B1
by Zhike Zhao, Jianghao Feng and Caizhang Wu
Biosensors 2025, 15(8), 538; https://doi.org/10.3390/bios15080538 - 15 Aug 2025
Viewed by 210
Abstract
In addressing the challenges posed by high costs, low accuracy, and cumbersome operations in mycotoxin detection, a novel aptamer-antibody sandwich chemiluminescent biosensor for detecting aflatoxin B1 (AFB1) was developed. The indirect competition between AFB1, aflatoxin B1-ovomucoid [...] Read more.
In addressing the challenges posed by high costs, low accuracy, and cumbersome operations in mycotoxin detection, a novel aptamer-antibody sandwich chemiluminescent biosensor for detecting aflatoxin B1 (AFB1) was developed. The indirect competition between AFB1, aflatoxin B1-ovomucoid complete antigen (AFB1-OVA), and rabbit anti-ovomucoid (OVA) antibody results in the formation of a sandwich complex. This sandwich assay is linked to a horseradish peroxidase-labeled antibody, which catalyzes luminol chemiluminescence for the indirect detection of AFB1. The biosensor was designed to operate with high precision, low cost, and a low detection limit for AFB1, which is contingent upon experimental conditions such as pH, reagent concentration, temperature, and incubation time. The optimization of pH, aptamer concentration, competitive incubation time, competitive incubation temperature, and HRP-labeled antibody concentration was instrumental in achieving these objectives. Experimental findings demonstrated that the sensor’s detection limit was 0.067 ng/mL, exhibiting excellent linearity (R2 = 0.99679) within the concentration range of 0.25–10 ng/mL. The recovery rate of spiked samples ranged from 94.4% to 108.05%. This sensor boasts a low detection limit, straightforward operation, and minimal cost, thus offering a novel solution for developing cost-effective, high-precision mycotoxin detection methods. Full article
(This article belongs to the Section Optical and Photonic Biosensors)
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21 pages, 4205 KiB  
Article
Safety Evaluation and Biodistribution of Fetal Umbilical Cord Mesenchymal Stem Cells-Derived Small Extracellular Vesicles in Sprague Dawley Rats
by Illayaraja Krishnan, Ubashini Vijakumaran, Ng Min Hwei, Law Jia Xian, Mohd Rafizul Mohd Yusof, Thavachelvi Thangarajah, Tan Geok Chin, Yin Ping Wong, Anusha Kalyanasundaram, Zalina Mahmood, Shathiya Rajamanickam, Baskar Subramani and Yogeswaran Lokanathan
Int. J. Mol. Sci. 2025, 26(14), 6806; https://doi.org/10.3390/ijms26146806 - 16 Jul 2025
Viewed by 564
Abstract
Umbilical cord mesenchymal stem cells (UCMSCs)-derived small extracellular vehicles (sEVs) are reported to offer therapeutic effects in regenerative medicine, but they lack safety and biodistribution profiles to support smooth translation at the clinical stage and regulatory requirements. Our study aimed to determine the [...] Read more.
Umbilical cord mesenchymal stem cells (UCMSCs)-derived small extracellular vehicles (sEVs) are reported to offer therapeutic effects in regenerative medicine, but they lack safety and biodistribution profiles to support smooth translation at the clinical stage and regulatory requirements. Our study aimed to determine the safety and biodistribution profile in a healthy animal model before application in the metabolic syndrome model. Method: Healthy male Sprague Dawley (SD) rats were given an intravenous (IV) injection of normal saline (control group) or pooled fetal UCMSCs-derived sEVs (treated group) every three weeks for 90 days. Morbidity and mortality observation (daily), physical measurements (weekly), selected serum biochemistry (every three weeks), and hematology (every three weeks) were performed for 90 days. Acute toxicity (on day 14) and sub-chronic toxicity (on day 90) were assessed for gross necropsy, relative organ weight, and histopathological assessment of lungs, liver, spleen, kidney, and lymph nodes. Separately, a biodistribution study was conducted with the sEVs preparations labeled with PKH26 fluorescent dye, given intravenously to the rats. The organs were harvested 24 h post-injection. There were no drastic changes in either group’s morbidity or mortality, physical, hematological, and biochemistry evaluation. The histopathological assessment concluded moderate (focal) inflammation in the treated group’s kidneys and signs of recovery from the inflammation and vascular congestion in the liver. A biodistribution study revealed a higher accumulation of sEVs in the spleen. Multiple IV injections of the pooled fetal UCMSCs-derived sEVs in healthy male SD rats were deemed safe. The sEVs were abundantly distributed in the spleen 24 h post-injection. Full article
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22 pages, 3733 KiB  
Article
Combating Traumatic Brain Injury: A Dual-Mechanism Hydrogel Delivering Salvianolic Acid A and Hydroxysafflor Yellow A to Block TLR4/NF-κB and Boost Angiogenesis
by Guoying Zhou, Yujia Yan, Linh Nguyen, Jiangkai Fan, Xiao Zhang, Li Gan, Tingzi Yan and Haitong Wan
Polymers 2025, 17(14), 1900; https://doi.org/10.3390/polym17141900 - 9 Jul 2025
Viewed by 572
Abstract
Traumatic brain injury (TBI) leads to severe neurological dysfunction, disability, and even death. Surgical intervention and neurorehabilitation represent the current clinical management methods, yet there remains no effective treatment for recovery after TBI. Post-traumatic hyperinflammation and vascular injury are the key therapeutic challenges. [...] Read more.
Traumatic brain injury (TBI) leads to severe neurological dysfunction, disability, and even death. Surgical intervention and neurorehabilitation represent the current clinical management methods, yet there remains no effective treatment for recovery after TBI. Post-traumatic hyperinflammation and vascular injury are the key therapeutic challenges. Therefore, a novel-designed multifunctional HT/SAA/HSYA hydrogel based on hyaluronic acid (HA) co-loaded with salvianolic acid A (SAA) and hydroxysafflor yellow A (HSYA) was developed in order to simultaneously target inflammation and vascular injury, addressing key pathological processes in TBI. The HT hydrogel was formed through covalent cross-linking of tyramine-modified HA catalyzed by horseradish peroxidase (HRP). Results demonstrated that the HT hydrogel possesses a porous structure, sustained release capabilities of loaded drugs, suitable biodegradability, and excellent biocompatibility both in vitro and in vivo. WB, immunofluorescence staining, and PCR results revealed that SAA and HSYA significantly reduced the expression level of pro-inflammatory cytokines (IL-1β and TNF-α) and inhibited M1 macrophage polarization through the suppression of the TLR4/NF-κB inflammatory pathway. In vivo experiments confirmed that the HT/SAA/HSYA hydrogel exhibited remarkable pro-angiogenic effects, as evidenced by increased expression of CD31 and α-SMA. Finally, H&E staining showed that the HT/SAA/HSYA hydrogel effectively reduced the lesion volume in a mouse TBI model, and demonstrated more pronounced effects in promoting brain repair at the injury site, compared to the control and single-drug-loaded hydrogel groups. In conclusion, the HT hydrogel co-loaded with SAA and HSYA demonstrates excellent anti-inflammatory and pro-angiogenic effects, offering a promising therapeutic approach for brain repair following TBI. Full article
(This article belongs to the Section Polymer Applications)
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20 pages, 861 KiB  
Article
A Longitudinal Ecologic Analysis of Neighborhood-Level Social Inequalities in Health in Texas
by Catherine Cubbin, Abena Yirenya-Tawiah, Yeonwoo Kim, Bethany Wood, Natasha Quynh Nhu Bui La Frinere-Sandoval and Shetal Vohra-Gupta
Int. J. Environ. Res. Public Health 2025, 22(7), 1076; https://doi.org/10.3390/ijerph22071076 - 5 Jul 2025
Viewed by 483
Abstract
Most health studies use cross-sectional data to examine neighborhood context because of the difficulty of collecting and analyzing longitudinal data; this prevents an examination of historical trends that may influence health outcomes. Using the Neighborhood Change Database, we categorized longitudinal (1990–2010) poverty and [...] Read more.
Most health studies use cross-sectional data to examine neighborhood context because of the difficulty of collecting and analyzing longitudinal data; this prevents an examination of historical trends that may influence health outcomes. Using the Neighborhood Change Database, we categorized longitudinal (1990–2010) poverty and White concentration trajectories (long-term low, long-term moderate, long-term high, increasing, or decreasing) for Texas census tracts and linked them to tract-level health-related characteristics (social determinants of health [SDOH] in 2010, health risk and preventive behaviors [HRPB] in 2017, and health status/outcomes [HSO] in 2017) from multiple sources (N = 2961 tracts). We conducted univariate and bivariate descriptive analyses, followed by linear regressions adjusted for population density. SDOH, HRPB, and HSO measures varied widely across census tracts. Both poverty and White concentration trajectories were strongly and consistently associated with a wide range of SDOH. Long-term high-poverty and low-White tracts showed the greatest disadvantages, while long-term low-poverty and high-White tracts had the most advantages. Neighborhoods undergoing changes in poverty or White concentrations, either increasing or decreasing, had less advantageous SDOH compared with long-term low-poverty or long-term high-White neighborhoods. While associations between poverty, White concentration trajectories, and SDOH were consistent, those with HRPB and HSO were less so. Understanding impact of the relationships between longitudinal neighborhood poverty and racial/ethnic composition on health can benefit stakeholders designing policy proposals and intervention strategies. Full article
(This article belongs to the Special Issue 3rd Edition: Social Determinants of Health)
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23 pages, 11450 KiB  
Article
Inhibition Effects and Mechanism Study of rAj-HRP30, a Recombinant Histidine-Rich Peptide from Apostichopus japonicus, on the Viability of Pancreatic Ductal Adenocarcinoma Cells Panc01 and Panc02
by Yuyao Song, Shan Gao, Jingwei Jiang, Yuebin Zhang, Jingyu Zhang, Xiaona Wang, Li Lv, Zunchun Zhou and Jihong Wang
Int. J. Mol. Sci. 2025, 26(4), 1485; https://doi.org/10.3390/ijms26041485 - 11 Feb 2025
Viewed by 1146
Abstract
rAj-HRP30 is a recombinant peptide derived from the wild-type rAj-HRP of Apostichopus japonicus through a gene-shortening mutation. It has a high histidine content (53.3% in its primary structure) and a molecular weight of 3.919 kDa, classifying it as a histidine-rich peptide. The literature [...] Read more.
rAj-HRP30 is a recombinant peptide derived from the wild-type rAj-HRP of Apostichopus japonicus through a gene-shortening mutation. It has a high histidine content (53.3% in its primary structure) and a molecular weight of 3.919 kDa, classifying it as a histidine-rich peptide. The literature reports indicate that human histidine-rich peptides exhibit antitumor activity. Previous research by our group demonstrated similar properties in rAj-HRP, the precursor of rAj-HRP30. Therefore, this study used Panc01 (human) and Panc02 (mouse) cells—highly malignant models with limited targeted therapies—to investigate the antitumor activity and mechanisms of rAj-HRP30 and evaluate its potential for pancreatic cancer treatment. This study designed a gene-shortening strategy for rAj-HRP and artificially synthesized the gene sequence of rAj-HRP30. The cDNA sequence of rAj-HRP30 was cloned into the pET23b vector, and the recombinant plasmid pET23b-HRP30 was transformed into E. coli BL21 for expression. Following IPTG induction, the recombinant peptide was purified using nickel ion affinity chromatography, yielding rAj-HRP30 with a purity exceeding 95%. rAj-HRP30 markedly inhibited the adhesion, migration, and invasion of Panc01 and Panc02 cells. It also disrupted cellular morphology and cytoskeletal structure while inducing apoptosis. These effects were dose-dependent. After confirming the in vitro anticancer activity of rAj-HRP30, this study employed Panc02 cells as a model to investigate its inhibitory mechanisms using Western blot analysis. The results revealed that rAj-HRP30 reduced FGFR1 expression in Panc02 cells and inhibited the downstream FYN and FAK signaling pathways, subsequently blocking the PI3K/AKT signaling and apoptosis pathways. In the apoptotic pathway, rAj-HRP30 was able to downregulate the expression of Bcl-2, Caspase-9, Caspase-3, Caspase-7, and PARP1 and upregulate the expression of Bax, cleaved Caspase-9, cleaved Caspase-3, cleaved Caspase-7, and cleaved-PARP1 to induce apoptosis in Panc02 cells. Furthermore, rAj-HRP30 also downregulated the expression of MMP2 and MMP9, thereby inhibiting the migration and invasion of Panc02 cells. Conclusion: rAj-HRP30 exhibits significant inhibitory effects on pancreatic ductal adenocarcinoma Panc01 and Panc02 cells in vitro. Its mechanism involves FGFR1-related signaling and apoptosis pathways. rAj-HRP30 shows promise as a therapeutic agent targeting FGFR for pancreatic cancer. Full article
(This article belongs to the Special Issue Oxidative Stress and Autophagy in Cancer Cells)
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11 pages, 2775 KiB  
Article
Detection of Aflatoxin B1 in Wheat Based on Nucleic Aptamer Chemiluminescence Sensor
by Zebing Zhang, Caizhang Wu and Zhike Zhao
Sensors 2025, 25(4), 988; https://doi.org/10.3390/s25040988 - 7 Feb 2025
Cited by 3 | Viewed by 926
Abstract
In this study, we developed a low-cost, high-sensitivity chemiluminescence competitive aptamer sensor for the detection of aflatoxin B1 (AFB1) in wheat samples. The optical fiber sensor was self-made, and it utilized biotin and streptavidin (SA) link aptamer and horseradish peroxidase [...] Read more.
In this study, we developed a low-cost, high-sensitivity chemiluminescence competitive aptamer sensor for the detection of aflatoxin B1 (AFB1) in wheat samples. The optical fiber sensor was self-made, and it utilized biotin and streptavidin (SA) link aptamer and horseradish peroxidase (HRP) for the chemiluminescence detection, achieving competitive assay between the AFB1 and AFB1 antigen. We adjusted the experimental conditions of the sensor base on the date of optimization of the experimental conditions and chose coated antigens on the surface of carboxyl magnetic particles. Under conditions optimized by testing key parameters, the assay results showed that the chemiluminescence intensity and AFB1 concentration demonstrated a strong linear relationship (R2 = 0.995), the dynamic range was from 0.1 to 10 ng/mL with a detection limit of 0.09 ng/mL, and the aptamer exhibited good specificity and anti-interference ability. Testing the wheat samples showed that the spiked recovery rate ranged from 79.19% to 113.21%. The sensor possesses characteristics of low detection limits, simple manufacturing methods, and affordability, providing a novel solution for the development of low-cost and high-sensitivity AFB1 detection equipment. Full article
(This article belongs to the Section Biomedical Sensors)
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20 pages, 7629 KiB  
Article
MgAl-Layered Double Hydroxide-Coated Bio-Silica as an Adsorbent for Anionic Pollutants Removal: A Case Study of the Implementation of Sustainable Technologies
by Muna Abdualatif Abduarahman, Marija M. Vuksanović, Nataša Knežević, Katarina Banjanac, Milena Milošević, Zlate Veličković and Aleksandar Marinković
Int. J. Mol. Sci. 2024, 25(21), 11837; https://doi.org/10.3390/ijms252111837 - 4 Nov 2024
Cited by 2 | Viewed by 1651
Abstract
The adsorption efficiency of Cr(VI) and anionic textile dyes onto MgAl-layered double hydroxides (LDHs) and MgAl-LDH coated on bio-silica (b-SiO2) nanoparticles (MgAl-LDH@SiO2) derived from waste rice husks was studied in this work. The material was characterized using field-emission scanning [...] Read more.
The adsorption efficiency of Cr(VI) and anionic textile dyes onto MgAl-layered double hydroxides (LDHs) and MgAl-LDH coated on bio-silica (b-SiO2) nanoparticles (MgAl-LDH@SiO2) derived from waste rice husks was studied in this work. The material was characterized using field-emission scanning electron microscopy (FE-SEM/EDS), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), and X-ray photoelectron spectroscopic (XPS) techniques. The adsorption capacities of MgAl-LDH@SiO2 were increased by 12.2%, 11.7%, 10.6%, and 10.0% in the processes of Cr(VI), Acid Blue 225 (AB-225), Acid Violet 109 (AV-109), and Acid Green 40 (AG-40) dye removal versus MgAl-LDH. The obtained results indicated the contribution of b-SiO2 to the development of active surface functionalities of MgAl-LDH. A kinetic study indicated lower intraparticle diffusional transport resistance. Physisorption is the dominant mechanism for dye removal, while surface complexation dominates in the processes of Cr(VI) removal. The disposal of effluent water after five adsorption/desorption cycles was attained using enzymatic decolorization, photocatalytic degradation of the dyes, and chromate reduction, satisfying the prescribed national legislation. Under optimal conditions and using immobilized horseradish peroxidase (HRP), efficient decolorization of effluent solutions containing AB-225 and AV-109 dyes was achieved. Exhausted MgAl-LDH@SiO2 was processed by dissolution/precipitation of Mg and Al hydroxides, while residual silica was used as a reinforcing filler in polyester composites. The fire-proofing properties of composites with Mg and Al hydroxides were also improved, which provides a closed loop with zero waste generation. The development of wastewater treatment technologies and the production of potentially marketable composites led to the successful achievement of both low environmental impacts and circular economy implementation. Full article
(This article belongs to the Section Materials Science)
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28 pages, 5954 KiB  
Article
Endophenotypes of Primary Osteoarthritis of the Hip Joint in the Bulgarian Population over 60 Years Old
by Lyubomir Sapundzhiev, Tanya Sapundzhieva, Kamen Klinkanov, Martin Mitev, Kiril Simitchiev and Anastas Batalov
Life 2024, 14(5), 622; https://doi.org/10.3390/life14050622 - 11 May 2024
Cited by 1 | Viewed by 2046
Abstract
Aim. To identify subgroups of patients with primary osteoarthritis of the hip joint (pHOA) with similar imaging and laboratory findings, disease evolution, and response to conventional therapies. Methods. We performed further statistical analyses on patient data from two published, double-blind, randomized, and placebo-controlled [...] Read more.
Aim. To identify subgroups of patients with primary osteoarthritis of the hip joint (pHOA) with similar imaging and laboratory findings, disease evolution, and response to conventional therapies. Methods. We performed further statistical analyses on patient data from two published, double-blind, randomized, and placebo-controlled studies (DB-RCTs), which examined the effects of intra-articular corticosteroids (ia-CSs), hyaluronic acid (ia-HA)—KИ-109-3-0008/14.01.2014, and intravenous bisphosphonates (iv-BPs) -KИ- 109-3-0009/14.01.2014 compared to the country’s standard pHOA therapy. The data span an 8-year follow-up of 700 patients with pHOA, including: 1. Clinical parameters (WOMAC-A, B, C, and T; PtGA). 2. Laboratory markers (serum calcium and phosphate levels; 25-OH-D and PTH, markers for bone sCTX-I and cartilage uCTX-II turnover). 3. Radiological indicators: X-ray stage (Kellgren-Lawrence (K/L) and model (Bombelli/OOARSI), width (mJSW), speed (JSN mm/year), and zone of maximum narrowing of the joint space (max-JSN)—determining the type of femoral head migration (FHM). 4. DXA indicators: bone geometry (HAL; NSA; and MNW); changes in regional and total bone mineral density (TH-BMD, LS-BMD, and TB-BMD). 5. Therapeutic responses (OARSI/MCII; mJSW; JSNmm/yearly) to different drug regimens (iv-BP -zoledronic acid (ZA/-5 mg/yearly for 3 years)); ia-CS 40 mg methylprednisolone acetate, twice every 6 months; and ia-HA with intermediate molecular weight (20 mg/2 mL × 3 weekly applications, two courses every 6 months) were compared to standard of care therapy (Standard of Care/SC/), namely D3-supplementation according to serum levels (20–120 ng/mL; target level of 60 ng/mL), simple analgesics (paracetamol, up to 2.0 g/24 h), and physical exercises. The abovementioned data were integrated into a non-supervised hierarchical agglomerative clustering analysis (NHACA) using Ward’s linkage method and the squared Euclidean distance to identify different endophenotypes (EFs). Univariate and multivariate multinomial logistic regression analyses were performed to determine the impact of sex and FHM on clinical and radiographic regression of pHOA. Results. A baseline cluster analysis using incoming (M0) patient data identified three EFs: hypertrophic H-HOA, atrophic A-HOA, and intermediate I-HOA. These EFs had characteristics that were similar to those of patients grouped by radiographic stage and pattern (‘H’-RPs, ‘I’-RPs, and ‘A’-RPs), p < 0.05). The repeated cluster analysis of M36 data identified four EF pHOAs: 1. Hypertrophic (slow progressors, the influence of the type of femoral head migration (FHM) outweighing the influence of sex on progression), progressing to planned total hip replacement (THR) within 5 (K/LIII) to 10 (K/LII) years. 2. Intermediate (sex is more important than the FHM type for progression) with two subgroups: 2#: male-associated (slow progressors), THR within 4 (K/LIII) to 8 years. (K/LII). 2* Female-associated (rapid progressors), THR within 3 (K/LIII) to 5 (K/LII) years. 3. Atrophic (rapid progressors; the influence of FHM type outweighs that of sex), THR within 2 (K/LIII) to 4 (K/LII) years. Each EF, in addition to the patient’s individual progression rate, was also associated with a different response to the aforementioned therapies. Conclusions. Clinical endophenotyping provides guidance for a personalized approach in patients with pHOA, simultaneously assisting the creation of homogeneous patient groups necessary for conducting modern genetic and therapeutic scientific studies. Full article
(This article belongs to the Section Medical Research)
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18 pages, 7989 KiB  
Article
Injectable Hydrogels Based on Hyaluronic Acid and Gelatin Combined with Salvianolic Acid B and Vascular Endothelial Growth Factor for Treatment of Traumatic Brain Injury in Mice
by Guoying Zhou, Yajie Cao, Yujia Yan, Haibo Xu, Xiao Zhang, Tingzi Yan and Haitong Wan
Molecules 2024, 29(8), 1705; https://doi.org/10.3390/molecules29081705 - 10 Apr 2024
Cited by 11 | Viewed by 3173
Abstract
Traumatic brain injury (TBI) leads to structural damage in the brain, and is one of the major causes of disability and death in the world. Herein, we developed a composite injectable hydrogel (HA/Gel) composed of hyaluronic acid (HA) and gelatin (Gel), loaded with [...] Read more.
Traumatic brain injury (TBI) leads to structural damage in the brain, and is one of the major causes of disability and death in the world. Herein, we developed a composite injectable hydrogel (HA/Gel) composed of hyaluronic acid (HA) and gelatin (Gel), loaded with vascular endothelial growth factor (VEGF) and salvianolic acid B (SAB) for treatment of TBI. The HA/Gel hydrogels were formed by the coupling of phenol-rich tyramine-modified HA (HA-TA) and tyramine-modified Gel (Gel-TA) catalyzed by horseradish peroxidase (HRP) in the presence of hydrogen peroxide (H2O2). SEM results showed that HA/Gel hydrogel had a porous structure. Rheological test results showed that the hydrogel possessed appropriate rheological properties, and UV spectrophotometry results showed that the hydrogel exhibited excellent SAB release performance. The results of LIVE/DEAD staining, CCK-8 and Phalloidin/DAPI fluorescence staining showed that the HA/Gel hydrogel possessed good cell biocompatibility. Moreover, the hydrogels loaded with SAB and VEGF (HA/Gel/SAB/VEGF) could effectively promote the proliferation of bone marrow mesenchymal stem cells (BMSCs). In addition, the results of H&E staining, CD31 and α-SMA immunofluorescence staining showed that the HA/Gel/SAB/VEGF hydrogel possessed good in vivo biocompatibility and pro-angiogenic ability. Furthermore, immunohistochemical results showed that the injection of HA/Gel/SAB/VEGF hydrogel to the injury site could effectively reduce the volume of defective tissues in traumatic brain injured mice. Our results suggest that the injection of HA/Gel hydrogel loaded with SAB and VEGF might provide a new approach for therapeutic brain tissue repair after traumatic brain injury. Full article
(This article belongs to the Special Issue Hydrogels: Preparation, Characterization, and Applications)
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17 pages, 8531 KiB  
Article
Antibacterial Activity and Mechanism of Three Root Exudates from Mulberry Seedlings against Ralstonia pseudosolanacearum
by Ping Li, Siyi Wang, Mengyuan Liu, Xue Dai, Huicong Shi, Weihong Zhou, Sheng Sheng and Fuan Wu
Plants 2024, 13(4), 482; https://doi.org/10.3390/plants13040482 - 8 Feb 2024
Cited by 10 | Viewed by 2712
Abstract
Bacterial wilt is a significant soil-borne disease that poses a threat to mulberry production yield and quality of agricultural production worldwide. However, the disease resistance mechanisms dependent on root exudates are not well understood. In this present study, we investigated the antibacterial mechanisms [...] Read more.
Bacterial wilt is a significant soil-borne disease that poses a threat to mulberry production yield and quality of agricultural production worldwide. However, the disease resistance mechanisms dependent on root exudates are not well understood. In this present study, we investigated the antibacterial mechanisms of the main active substances (erucamide, oleamide, and camphor bromide) present in mulberry root exudates (MRE) against Ralstonia pseudosolanacearum (Rp), the causal agent of bacterial wilt. Our findings revealed that these three active substances inhibited the growth activity of Rp by affecting the cell morphology and extracellular polysaccharide content, as well as triggering a burst of reactive oxygen species. The active substances induced oxidative stress, leading to a decrease in Rp growth. Additionally, the expression levels of key genes in the hrp gene cluster (hrpB, hrpX, and hrpF) and other virulence-related genes (such as ripAW, ripAE, Rs5-4819, Rs5-4374, ace, egl3, and pehB) were significantly reduced upon treatment with the active substances. Further pathogenicity experiments demonstrated that root exudates (at a concentration of 1.5 mg·mL−1) delayed or slowed down the occurrence of bacterial wilt in mulberry. These findings provide valuable insight into the antimicrobial mechanisms of MRE against Rp and lay a theoretical foundation for the development and application of biocontrol agents to control mulberry bacterial wilt. Full article
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13 pages, 1524 KiB  
Article
Release of Pro-Inflammatory/Angiogenic Factors by Retinal Microvascular Cells Is Mediated by Extracellular Vesicles Derived from M1-Activated Microglia
by Elena Beltramo, Aurora Mazzeo and Massimo Porta
Int. J. Mol. Sci. 2024, 25(1), 15; https://doi.org/10.3390/ijms25010015 - 19 Dec 2023
Cited by 7 | Viewed by 2973
Abstract
The interactions between the neuronal and vascular sides of the retina during diabetic retinopathy (DR) have gained increasing attention. Microglia is responsible for the immune response to inflammation inside the retina, which could be mediated by paracrine signals carried by extracellular vesicles (EVs). [...] Read more.
The interactions between the neuronal and vascular sides of the retina during diabetic retinopathy (DR) have gained increasing attention. Microglia is responsible for the immune response to inflammation inside the retina, which could be mediated by paracrine signals carried by extracellular vesicles (EVs). We aimed to characterize EVs released from immortalized human microglial cells in inflammation and investigate their effects on the retinal microvasculature and the anti-inflammatory potential of thiamine in this context. M1 pro-inflammatory polarization in microglia was induced through a cytokine cocktail. EVs were isolated from the supernatants, characterized, and used to stimulate human retinal endothelial cells (HRECs) and pericytes (HRPs). Microvascular cell functions and their release of pro-inflammatory/angiogenic factors were assessed. M1-derived EVs showed increased content of miR-21, miR-155, CCL2, MMP2, and MMP9, and enhanced apoptosis, proliferation, migration, and ROS production in HRPs and HRECs. IL-1β, IL-6, MMP9, CCL2, and VEGF release increased in HRPs exposed to M1-derived EVs, while HRECs showed augmented IL-6, Ang2, VEGF, and PDFG-B. Addition of thiamine to M1-microglial cultures reverted most of these effects. In conclusion, M1-derived EVs stimulate functional changes and secretion of pro-inflammatory/angiogenic molecules in microvascular cells, exacerbating inflammatory damage and retinopathy features. Thiamine added to microglia exerts anti-inflammatory effects. Full article
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28 pages, 2229 KiB  
Article
Clonal Neoantigen: Emerging “Mechanism-based” Biomarker of Immunotherapy Response
by John Nemunaitis, Laura Stanbery, David Willoughby, Ernest Bognar, Scott Brun, Adam Walter, Bradley J. Monk, Rodney P. Rocconi, Khalil Choucair and Robert L. Coleman
Cancers 2023, 15(23), 5616; https://doi.org/10.3390/cancers15235616 - 28 Nov 2023
Cited by 6 | Viewed by 3928
Abstract
Clonal mutations represent the initiating molecular defects related to cellular transition of a normal phenotype to a malignant phenotype. Molecular genomic assessment utilizing next generation and whole exome sequencing is now being increasingly applied to biomarker determination to refine the use of targeted [...] Read more.
Clonal mutations represent the initiating molecular defects related to cellular transition of a normal phenotype to a malignant phenotype. Molecular genomic assessment utilizing next generation and whole exome sequencing is now being increasingly applied to biomarker determination to refine the use of targeted immune therapies. Case examples followed by retrospective study assessment have convincingly demonstrated clonal neoantigens provide a relevant predictor of response to checkpoint inhibition. A meta-analysis, by Litchfield et al., of over 1000 cancer patients from 12 landmark trials demonstrated no clinical benefit to checkpoint inhibitor (CPI) therapy in correlation to high subclonal tumor mutational burden (TMB), whereas high clonal TMB was found to be significantly correlated with better overall survival (p = 0.000000029). We discuss the mechanism of clonal vs. subclonal neoantigen targeting relationship to homologous recombination proficient (HRP) profile, evidence of preclinical and clinical benefit related to clonal neoantigens, and review a novel developing therapy called Vigil®, designed to expand the clonal neoantigen targeting effector cell populations. Vigil® is an autologous cellular immunotherapy which is designed to carry the full set of personal clonal neoantigens. Phase 2b results demonstrate a durable recurrence-free survival (RFS) and overall survival (OS) advantage for Vigil® in a subset ovarian cancer population with an HRP cancer profile. Full article
(This article belongs to the Collection Oncology: State-of-the-Art Research in the USA)
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16 pages, 2523 KiB  
Article
Ralstonia solanacearum Suppresses Tomato Root Growth by Downregulation of a Wall-Associated Receptor Kinase
by Sushuang Liu, Qi Xue, Shuying Zhu, Yanmin Liu and Huasong Zou
Plants 2023, 12(20), 3600; https://doi.org/10.3390/plants12203600 - 17 Oct 2023
Cited by 3 | Viewed by 2379
Abstract
The root architecture of a range of host plants is altered in response to Ralstonia solanacearum infection. This work aimed to identify host genes involved in root development during R. solanacearum infection. A deficient mutant of the type III secretion system regulator hrpB [...] Read more.
The root architecture of a range of host plants is altered in response to Ralstonia solanacearum infection. This work aimed to identify host genes involved in root development during R. solanacearum infection. A deficient mutant of the type III secretion system regulator hrpB was created in R. solanacearum GMI1000. The hrpB mutant was impaired in virulence but showed a similar suppressive effect as wild-type GMI1000 on tomato root development. Based on comparative transcriptome analysis, 209 genes were found that showed the same changed expression pattern in GMI1000 and hrpB mutant infected roots relative to uninoculated roots. Among them, the wall-associated receptor kinase WAKL20 was substantially downregulated in GMI1000 and hrpB mutant infected roots. Knockdown of WAKL20 led to a shorter primary root length and fewer lateral roots in tomato as well as in Nicotiana benthamiana. The WAKL20 is a pivotal target suppressed by R. solanacearum to shape the altered root development during infection. Full article
(This article belongs to the Special Issue Adaptation of Mutualistic Plant-Microbe Systems to Abiotic Stresses)
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13 pages, 2420 KiB  
Article
Unraveling the Antioxidant Activity of 2R,3R-dihydroquercetin
by Yaping Xu, Zhengwen Li, Yue Wang, Chujie Li, Ming Zhang, Haiming Chen, Wenxue Chen, Qiuping Zhong, Jianfei Pei, Weijun Chen, Guido R. M. M. Haenen and Mohamed Moalin
Int. J. Mol. Sci. 2023, 24(18), 14220; https://doi.org/10.3390/ijms241814220 - 18 Sep 2023
Cited by 6 | Viewed by 2224
Abstract
It has been reported that in an oxidative environment, the flavonoid 2R,3R-dihydroquercetin (2R,3R-DHQ) oxidizes into a product that rearranges to form quercetin. As quercetin is a very potent antioxidant, much better than 2R,3R-DHQ, [...] Read more.
It has been reported that in an oxidative environment, the flavonoid 2R,3R-dihydroquercetin (2R,3R-DHQ) oxidizes into a product that rearranges to form quercetin. As quercetin is a very potent antioxidant, much better than 2R,3R-DHQ, this would be an intriguing form of targeting the antioxidant quercetin. The aim of the present study is to further elaborate on this targeting. We can confirm the previous observation that 2R,3R-DHQ is oxidized by horseradish peroxidase (HRP), with H2O2 as the oxidant. However, HPLC analysis revealed that no quercetin was formed, but instead an unstable oxidation product. The inclusion of glutathione (GSH) during the oxidation process resulted in the formation of a 2R,3R-DHQ-GSH adduct, as was identified using HPLC with IT-TOF/MS detection. GSH adducts appeared on the B-ring of the 2R,3R-DHQ quinone, indicating that during oxidation, the B-ring is oxidized from a catechol to form a quinone group. Ascorbate could reduce the quinone back to 2R,3R-DHQ. No 2S,3R-DHQ was detected after the reduction by ascorbate, indicating that a possible epimerization of 2R,3R-DHQ quinone to 2S,3R-DHQ quinone does not occur. The fact that no epimerization of the oxidized product of 2R,3R-DHQ is observed, and that GSH adducts the oxidized product of 2R,3R-DHQ on the B-ring, led us to conclude that the redox-modulating activity of 2R,3R-DHQ quinone resides in its B-ring. This could be confirmed by chemical calculation. Apparently, the administration of 2R,3R-DHQ in an oxidative environment does not result in ‘biotargeting’ quercetin. Full article
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71 pages, 483 KiB  
Perspective
Circuits and Biomarkers of the Central Nervous System Relating to Astronaut Performance: Summary Report for a NASA-Sponsored Technical Interchange Meeting
by Joshua S. Alwood, Ajitkumar P. Mulavara, Janani Iyer, Siddhita D. Mhatre, Susanna Rosi, Mark Shelhamer, Catherine Davis, Christopher W. Jones, Xiao Wen Mao, Rajeev I. Desai, Alexandra M. Whitmire and Thomas J. Williams
Life 2023, 13(9), 1852; https://doi.org/10.3390/life13091852 - 31 Aug 2023
Cited by 5 | Viewed by 3061
Abstract
Biomarkers, ranging from molecules to behavior, can be used to identify thresholds beyond which performance of mission tasks may be compromised and could potentially trigger the activation of countermeasures. Identification of homologous brain regions and/or neural circuits related to operational performance may allow [...] Read more.
Biomarkers, ranging from molecules to behavior, can be used to identify thresholds beyond which performance of mission tasks may be compromised and could potentially trigger the activation of countermeasures. Identification of homologous brain regions and/or neural circuits related to operational performance may allow for translational studies between species. Three discussion groups were directed to use operationally relevant performance tasks as a driver when identifying biomarkers and brain regions or circuits for selected constructs. Here we summarize small-group discussions in tables of circuits and biomarkers categorized by (a) sensorimotor, (b) behavioral medicine and (c) integrated approaches (e.g., physiological responses). In total, hundreds of biomarkers have been identified and are summarized herein by the respective group leads. We hope the meeting proceedings become a rich resource for NASA’s Human Research Program (HRP) and the community of researchers. Full article
(This article belongs to the Special Issue Current Challenges in Human Space Flight)
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