Search Results (335)

The leaf diseases of Panax notoginseng (Panax notoginseng (Burk) F. H. Chen) are mainly spread by spores. To enable rapid and sensitive detection of spores for early warning of disease spread, we developed a carbon dot-based fluorescent probe encapsulated by MIL-101 using wax apple as a green carbon source (WA-CDs@MIL-101). The WA-CDs@MIL-101 was thoroughly characterized, and the detection conditions were optimized. The interaction mechanism between WA-CDs@MIL-101 and spores was investigated. The fluorescence of WA-CDs@MIL-101 was recovered due to electrostatic adsorption between spores and WA-CDs@MIL-101. Under the optimized detection conditions, the probe exhibited excellent sensing performance, showing a strong linear relationship (R² = 0.9978) between spore concentration (0.0025–5.0 mg/L) and fluorescence recovery ratio, with a detection limit of 5.15 μg/L. The WA-CDs@MIL-101 was successfully applied to detect spores on Panax notoginseng leaves, achieving satisfactory recoveries (94–102%) with relative standard deviations of 1.3–3.4%. The WA-CDs@MIL-101 shows great promise for detecting spores on Panax notoginseng leaves. Full article

Monkeypox virus (MPXV) has caused 148,892 confirmed cases and 341 deaths from 137 countries worldwide, as reported by the World Health Organization (WHO), highlighting the urgent need for effective vaccines to prevent the spread of MPXV. Traditional vaccine development is low-throughput, expensive, time consuming, and susceptible to reversion to virulence. Alternatively, a reverse vaccinology approach offers a rapid, efficient, and safer alternative for MPXV vaccine design. Here, MPXV proteins associated with viral infection were analyzed for immunogenic epitopes to design multi-epitope vaccines based on B-cell, CD4+, and CD8+ epitopes. Epitopes were selected based on allergenicity, antigenicity, and toxicity parameters. The prioritized epitopes were then combined via peptide linkers and N-terminally fused to various protein adjuvants, including PADRE, beta-defensin 3, 50S ribosomal protein L7/12, RS-09, and the cholera toxin B subunit (CTB). All vaccine constructs were computationally validated for physicochemical properties, antigenicity, allergenicity, safety, solubility, and structural stability. The three-dimensional structure of the selected construct was also predicted. Moreover, molecular docking and molecular dynamics (MD) simulations between the vaccine and the TLR-4 immune receptor demonstrated a strong and stable interaction. The vaccine construct was codon-optimized for high expression in the E. coli and was finally cloned in silico into the pET21a (+) vector. Collectively, these results could represent innovative tools for vaccine formulation against MPXV and be transformative for other infectious diseases. Full article

High-grade serous ovarian cancer (HGSOC) is an aggressive gynecological malignancy characterized by intraperitoneal spread and chemotherapy resistance. Chemotherapies have demonstrated limited effectiveness in HGSOC, underscoring the urgent need to evaluate how the tumor microenvironment (TME) was reshaped by chemotherapy in different sites of tumor foci. In this study, we performed single-cell transcriptomic analysis to explore the TME in samples obtained from various sites of tumor foci, with or without the history of Neoadjuvant chemotherapy (NACT). We discovered that chemotherapy reshaped the tumor immune microenvironment, evident through the reduction in human leukocyte antigen (HLA) diversity and the increase in PDCD1/CD274 in CD8_ANXA1, LAMP3+ dendritic cell (DC_LAMP3), and EREG+ monocytes (mono_EREG). Moreover, cancer.cell.2, cancer-associated C3+ fibroblasts (CAF_C3), and Fibrocyte_CD34, which are prone to accumulate in the metastatic site and post-NACT group, harbored poor clinical outcome, reflected in the immune exclusion and tumor progression signaling. Cell–cell communication identified a stronger interaction between cancer.cell.2 and CAF_C3, as well as Fibrocyte_CD34, in post-NACT samples, indicating that chemotherapy reshapes pre-existing cell clusters in a site-dependent manner. Our findings suggest that chemotherapy and sites of foci were critical for the transcriptional reprogramming of pre-existed cell clusters. Our study offers a single-cell phenotype data substrate from which to develop a personalized combination of chemotherapy and immunotherapy. Full article

Background: hMPXV poses a major public health risk due to its human-to-human transmissibility, severe complications, especially in immunocompromised individuals, and global spread, necessitating effective surveillance and stringent prophylactic measures to mitigate its colossal impact. Objective: The study aimed to annotate hMPXV(IMV) proteins to propose a potential reverse vaccinology-based vaccine against hMPXV. Methods: The target MPXV(IMV) protein’s sequences, formatted in FASTA, were sourced from genome/proteome databases (BV-BRC and UniProt) (accessed on 6 November 2024), followed by CD-Hit-based redundancy removal. Epitope prediction for B-cells (lymphocytes), cytotoxic T-cells or cytotoxic T-lymphocytes (CTLs), and helper T-cells (HTLs) was executed using ABCpred, IEDB’s ANNs 4.0, and an artificial neural network-based alignment tool (NN-align 2.3)/ML-based tool (NetMHCII 2.3). Various immunoinformatics filters (antigenicity, toxicity, and allergenicity) were applied to substantiate the potency and safety of the formulated vaccine candidate. The constructed vaccine’s physiochemical and structural features (secondary and tertiary), with structural stability (confirmed by molecular docking followed by dynamic simulation with TLRs (TLR4 & TLR2) and MHCs), were determined. Additionally, cloning (using pET-28a(+) vector) was conducted to verify the vaccine’s expression potential and translation efficiency. The construct’s population coverage was also ascertained. Results: The MPXV-2-Beta vaccine constructs, of the six initially designed constructs, was identified as the most promising candidate, signifying nonallergenic profile and nontoxic features, with a predicted antigenicity score (PAS) = 0.7202, 407 residues, a molecular weight of 43,102.1 Da, pI of 9.2, and favorable stability parameters (AI: 65.65, GRAVY: −0.597, I-i: 25.92). It showed high solubility (score: 0.942). The ProSA Z-score of −9.38 confirmed the structural stability, reliability, and precision of the MPXV-2-Beta 3D model, which is comparable to experimental structures. Furthermore, 98.8% of all the residues nested within favored or allowed regions in a critical Ramachandran plot signified the model’s exceptional structural integrity and quality. Docking and dynamic simulation of MPXV-2-Beta with TLRs (TLR4 & TLR2) and MHCs demonstrated stiffer docking stability (strong polar and nonpolar interaction) and negative eigenvalue value (during dynamic simulation), suggesting its ability to enhance immune receptor activation under physiological conditions. MPXV-2-Beta was predicted to trigger a robust immune response (IR) with comprehensive world population coverage (98.55%, SD = 10.41). Conclusions: Based on the evaluated parameters, the MPXV-2-Beta designed in this study exhibited significant potential as an effective candidate against hMPXV. This study establishes a foundation for developing an efficient vaccine against hMPXV, requiring further experimental and clinical validation to confirm computational findings. Full article

Animal-based food is significant for human nutrition, as it represents an easily digestible source of high-quality proteins, fatty acids, fat-soluble vitamins, and energy. During evolution, humans developed anatomical, metabolic, and biochemical adaptations in the digestive tract, becoming increasingly dependent on nutritionally valuable food, such as animal-based products. Animal-based food can be a source of chemical substances that are harmful to health, such as contaminants (heavy metals, mycotoxins, organochlorine pesticides, etc.), veterinary drug residues, and additives. The subject of this paper is the determination of the content of essential (copper—Cu, iron—Fe, and chromium—Cr) and heavy and toxic metals (lead—Pb, cadmium—Cd, arsenic—As, and tin—Sn), as well as nitrites, in meat products, particularly pâtés and meat spreads available on the market in the Republic of Srpska. Determination of the content of metals was performed using the ICP-OES method, while nitrites were analyzed using the standard SRPS ISO 2918/1999 method. The obtained results indicate that the content of heavy and toxic metals and nitrites is lower than the maximum prescribed by national regulations, namely the Rulebook on Maximum Amounts of Certain Contaminants (Official Gazette of BA, No. 68/14, 79/16, 84/18) and the Rulebook on Additives in Food (Official Gazette of BA, No. 33/18 and 6/21). Full article

CD300 family members are immunoglobulin superfamily receptors that regulate immune cell function through either activating or inhibitory signals. Among them, CD300a is a prototypical inhibitory receptor, highly expressed in both myeloid and lymphoid lineages, and plays a pivotal role in the pathogenesis of inflammation and tumor immunity. CD300a transduces inhibitory signals in several immune cells—including mast cells, eosinophils, monocytes, dendritic cells (DCs), neutrophils, and natural killer (NK) cells—by recruiting SHP-1 phosphatase to immunoreceptor tyrosine-based inhibitory motifs (ITIMs) and suppressing activation pathways such as Toll-like receptor (TLR)-MyD88 and FcεRI signaling. Recent studies suggest that tumor cells may hijack CD300a-associated pathways to establish an immunosuppressive microenvironment that facilitates immune evasion, tumor survival, and potentially metastatic spread. Proposed mechanisms include reduced DC-mediated type I interferon (IFN) production, diminished NK cell cytotoxicity, and negative regulation of mast cell– and eosinophil-dependent anti-tumor responses. Although some of these findings are derived from in vivo models, the cumulative evidence positions CD300a as a critical immune checkpoint in tumor-associated immune regulation. In addition to its established roles in hematologic malignancies—including chronic lymphocytic leukemia, acute lymphoblastic leukemia, and acute myeloid leukemia—CD300a has also been implicated in modulating tumor-associated immune responses in other pathological contexts. While most studies emphasize its immune cell–mediated effects, emerging evidence suggests that CD300a may directly influence tumor progression by regulating immune homeostasis, intracellular signaling, and tumor microenvironment interactions. Collectively, these findings establish CD300a as a pleiotropic immunoregulatory molecule in both hematologic and non-hematologic malignancies, underscoring the need to further explore its broader relevance and therapeutic potential in cancer immunology. Full article

Enterotoxigenic Escherichia coli (ETEC), a common intestinal pathogen, can colonize the intestines and induce diarrhea in piglets, which brings great economic losses to the swine industry. Antibiotics are recommended to the treatment for diarrhea caused by ETEC in weaned piglets. However, with the emergence and spread of multidrug-resistant ETEC, there is an urgent need to develop alternatives to antibiotics. Due to the unique antibacterial mechanism of targeting bacterial membranes, antimicrobial peptides (AMPs) are promising candidates. In this study, the activity of crude recombinant porcine β-defensin 2 (rPBD2) expressed in Pichia pastoris (P. pastoris) was measured in vitro. Mice infected with ETEC were orally administered 16, 8, and 4 AU crude rPBD2 for 7 consecutive days to evaluate its anti-infective activity in vivo. The results showed that in addition to broad antibacterial activity against Gram-positive and -negative bacteria, crude rPBD2 displayed high tolerance to temperatures ranging from 20 to 60 °C, a broad range of pH, trypsin, pepsin, and physiological concentrations of salts. In an ETEC-induced mouse model, the oral administration of crude rPBD2 decreased diarrhea scores and the intestinal/carcass ratio and alleviated body weight loss. Additionally, crude rPBD2 decreased bacterial loads in stools and the colon (HP group), and the levels of serum pro-inflammatory cytokines IL-6 (HP group) and TNF-α (HP and MP groups), and increased the villus height and the ratio of villus height to crypt depth (VH/CD) in the ileum (HP and MP groups). Our study provides a cost-effective way for PBD2 production and identifies it as a promising candidate to combat ETEC-induced infection. Full article

Background/Objectives: Microbial infections represent a significant threat to public health due to the emergence and spread of antimicrobial resistance. Adjunctive and alternative therapeutic strategies are explored to tackle this issue, including the use of natural or synthetic antimicrobial peptides. Previous research showed that antibody-derived peptides possess antimicrobial, antiviral, and immunomodulatory properties. This study aimed to characterize newly designed antibody-derived peptides and evaluate their effectiveness against representative strains of Staphylococcus aureus, including drug-resistant isolates. Methods: Colony-forming unit assays and confocal microscopy studies were performed to evaluate peptide activity against planktonic microbial cells. Cytotoxicity tests were performed on THP-1 human monocytic cells. Circular dichroism (CD) and nuclear magnetic resonance (NMR) were employed for the conformational characterization of peptides. Results: The half-maximal effective concentrations of the peptides against bacterial reference strains and drug-resistant isolates ranged from 0.17 to 18.05 µM, while cytotoxic effects were not observed against mammalian cells. A killing kinetics analysis and observation by confocal microscopy of the interaction between peptides and bacteria suggested a mechanism of action involving membrane perturbation. CD studies showed that all peptides predominantly exhibit a random coil arrangement in aqueous solution. NMR spectroscopy revealed that the most active peptide adopts a helical conformation in the presence of membrane mimetics. Conclusions: The structural characterization and evaluation of the newly designed peptides’ antimicrobial activity may lead to the selection of a candidate to be further studied to develop an alternative treatment against microbial infections caused by drug-resistant strains. Full article

Background/Objectives: Immunocytokines (ICKs) are antibody–cytokine fusion proteins that deliver cytokines directly to tumors to increase immune responses, which are otherwise absent or limited by the delivery of antibodies alone. Tumor-associated glycoprotein-72 (TAG72) is overexpressed in numerous solid tumors. Methods: An anti-TAG72-IL-2 fusion protein was expressed in mammalian cells and tested in vitro for binding and bioactivity, and in vivo in two models. Results: In vitro studies showed high antigen specificity against TAG-72-positive tumor cell lines and IL-2 activity in CD25 (IL-2R)-positive reporter cells. To study the anti-tumor activity of huCC49-IL-2 in an immunocompetent model, the TAG-72 expression was established in murine mammary and colorectal cells by transfection with murine st6galnac-I gene (mSTn). Four daily doses of anti-TAG72-IL-2 monotherapy for TAG-72-expressing orthotopic murine mammary tumors in immunocompetent mice resulted in minimal whole-body toxicity and significant tumor growth reduction mediated by tumor infiltration of IFNγ⁺ CD8⁺ T cells. When mammary tumors were pretreated with image-guided fractionated radiation therapy (IGRT) followed by anti-TAG72-IL-2 therapy, an improved tumor growth inhibition was observed along with an increased tumor infiltration of IFNγ⁺ CD8⁺ T cells and a significant reduction in Foxp3⁺ CD4⁺ cells. Anti-TAG72-IL-2 monotherapy in TAG-72⁺ colorectal tumors led to a significant tumor reduction but also cures (4/7), with a rejection of rechallenges with both TAG-72-positive and -negative MC38 cells, thus demonstrating evidence of immune memory and antigen spreading. Conclusions: antiTAG-72-IL-2 therapy showed strong anti-tumor effects driven by activated CD8⁺ T cells making it a promising approach to the treatment of TAG-72⁺ tumors. Full article

Clostridioides difficile (CD) is a Gram-positive, spore-forming anaerobic bacterium, usually transmitted through the fecal–oral route, that can result from direct person-to-person contact, exposure to contaminated environmental surfaces, or contact with the hands of colonized healthcare personnel. An increased number of infections, especially healthcare-associated, with this etiology has been observed in most countries. As a spore-forming organism, CD is resistant to alcohol formulations and is a challenge for chemical disinfection. The solution could be the supplementation of traditional disinfection with non-touch techniques, such as UV-C radiation. The adoption of UV-C as a supplementary disinfection method in hospitals has significantly increased since the COVID-19 pandemic. However, there are no current guidelines concerning the use of UV-C disinfection in hospitals. The aim of this study was to evaluate the effectiveness of UV-C irradiation in inactivating Clostridioides difficile from different types of surfaces in hospital settings. The study was based on laboratory tests evaluating the efficacy in eliminating three different C. difficile strains on carriers made of plastic, metal and glass after 10 min exposure to UV-C (wavelength, 253.7 nm). We observed a wide range of reductions in the C. difficile suspensions depending on the density of the carrier contamination, type of carrier, strains and the location of the carrier. The percentage reductions ranged from 0 to 100%, but the best results were observed for glass, with lower initial suspension density and carrier placement on a door frame. Statistically significant differences were only seen in different suspension densities. Our experiment was a continuation of the tests done for non-sporing bacteria and C. auris, and there were some interesting differences in C. difficile reflecting its biology, especially its sensitivity to an aerobic atmosphere during the sample drying. Although the elimination of C. difficile by UV-C radiation was confirmed in our experiment, it was lower than in the case of non-spore-forming bacteria. Thus, this method may be used in healthcare settings (hospitals) for improving environmental safety and preventing C. difficile spreading. Full article

An African Swine Fever (ASF) outbreak was first recorded in the Philippines in July 2019. Since then, the disease has spread across provinces in Luzon, Visayas, and Mindanao, causing severe economic consequences for the country’s swine industry. Here, we report the genome sequencing of ASF virus strains from outbreaks in several provinces of the Philippines between 2021 and 2023, using a long-read tiled amplicon sequencing approach. The coding-complete genomes generated ranged from 187,609 to 189,540 bp in length, with GC contents of 38.4% to 38.5%. Notably, a strain from the Bataan province had a 1.9 kb deletion at the 5′-end, affecting several coding regions. The strains were characterized using 13 genes and regions; namely the B646L gene, the CD2v serogroup, the central variable region (CVR) of the B602L gene, the intergenic region (IGR) between the I73R and I329L genes, the IGR between A179L and A137R, O174L, K145R, Bt/Sj, J268L, and ECO2, the multigene family (MGF) 505-5R, and the MGF 505-9R and 10R regions. The ASFV strains were mostly related to Asian and European p72 genotype II strains. Genetic profiling provides valuable information on the diversity of local strains of ASFV in the Philippines, which are useful for epidemiology, diagnostics, and vaccine development. Full article

Background/Objectives: Osteosarcoma is an aggressive bone malignancy with high metastatic potential to the lungs. CTCs, as seeds of metastasis, play an important role in the spread of this cancer, and, therefore, their isolation, culture, and gene expression analysis promises valuable insights into the progression and metastatic cascade of osteosarcoma. The aim of this study was to isolate and culture CTCs from osteosarcoma-bearing mice and compare their migration, radioresistance, and gene expression with their parental cell line. Methods: CTCs from LM8-inoculated mice were isolated and cultured. The gene expression of the CTC-derived cell lines was then compared to the parental cell line. Furthermore, a Transwell assay, a clonogenic assay after irradiation, and immunohistochemical stainings were used to compare the CTC-derived cell lines with the parental cell line. Results: The CTC-derived cell lines differed significantly in gene expression from their parental cell line. 361 differentially expressed genes were identified, among which GO and KEGG analysis revealed major differences in the expression of genes related to antigen processing and presentation and extracellular matrix constituents. In addition, the CTC-derived cell lines were observed to have a higher migratory capacity and comparable radioresistance compared to the parental cell line. CD44 expression was found to be conserved in CTC-derived cell lines. Conclusions: This study provides a comparison between CTC-derived and their parental cell lines in terms of gene expression, migration, and radioresistance. Our findings allow for further research in the field of osteosarcoma CTCs and their generation. Furthermore, the identified DEGs between CTCs and their parental cell line can serve as a reference point for targeted therapies against osteosarcoma CTCs. Full article

The human T-cell leukemia type-1 (HTLV-1) retrovirus establishes chronic life-long infection in a fraction of infected individuals associated with severe pathological conditions. Although the mechanism driving disease development is not fully understood, current evidence indicates the essential functions of viral regulatory proteins. Among these, the p13 protein has previously been shown to localize to the inner mitochondrial membrane in T cells, altering mitochondrial biology and T-cell function. While CD4⁺ T cells are the primary cell target of HTLV-1 infection, genomic viral DNA has also been detected in monocytes, macrophages, and dendritic cells, which orchestrate innate and adaptive immunity and play a critical role in protecting against virus-induce diseases by establishing the appropriate balance of pro and anti-inflammatory responses. Given the central role of mitochondria in monocyte differentiation, we investigated the effect of p13 in monocytes/macrophages and found that by localizing to mitochondria, p13 affects mitochondrial respiration. Moreover, we demonstrate that p13 expression affects macrophage polarization to favor the recruitment of CD4⁺ T cells, the primary target of the virus, potentially facilitating the spread of viral infection and the development of disease. Full article

The presence of oil slicks in the ocean presents significant environmental and regulatory challenges for offshore oil processing operations. During primary oil–water separation, produced water is discharged into the ocean, carrying residual oil, which is measured using the total oil and grease (TOG) method. The formation and spread of oil slicks are influenced by metoceanographic variables, including wind direction (WD), wind speed (WS), current direction (CD), current speed (CS), wind wave direction (WWD), and peak period (PP). In Brazil, regulatory limits impose sanctions on companies when oil slicks exceed 500 m in length, making accurate prediction of their occurrence and extent crucial for offshore operators. This study follows three main stages. First, the performance of five machine learning classification algorithms is evaluated, selecting the most efficient method based on performance metrics from a Brazilian company’s oil slick database. Second, the best-performing model is used to analyze the influence of metoceanographic variables and TOG levels on oil slick occurrence and detection probability. Finally, the third stage examines the extent of detected oil slicks to identify key contributing factors. The prediction results enhance decision-support frameworks, improving monitoring and mitigation strategies for offshore oil discharges. Full article

Background/Objectives: In recent years, efforts by the scientific community to elucidate the underlying mechanisms of clonal expansion and selection within tumors have led to the theory of “tumor ecosystems”, implicating, among other factors, the role of the microenvironment in therapy resistance and tumor progression. In this context, the contribution of the microenvironment in the development of multiple myeloma (MM) is being investigated, imparting great emphasis on continuous clonal evolution. This process gives rise to aggressive clones with the potential to spread to extramedullary sites, rendering any treatment strategy practically ineffective. This systematic review aimed to gather knowledge about the immune microenvironment (IME) of extramedullary plasma cell myeloma and the differences in immune synthesis between medullary and extramedullary disease (EMD). Methods: A search strategy according to PRISMA guidelines was conducted in seven databases, and six articles meeting the inclusion criteria were encompassed in the study. Results: Results obtained from molecular analysis as well as flow cytometry and immunofluorescence indicated profound genetic instability at EMD sites along with spatial and temporal heterogeneity of the IME, implying a possible correlation between them. Both genetic and microenvironment variability were notably greater in EMD compared to medullary disease. The establishment of an immunosuppressive microenvironment was the rule, with exhausted CD8+ and natural killer (NK) cells, M2 macrophages, and inactivated dendritic cells found co-localized with neoplastic plasma cells, whereas cytotoxic CD8+ cells, M1 macrophages, and active dendritic cells congregated in tumor-free areas. Post-therapy alterations in the immune milieu were also noted and were concerned mostly the percentages of Tregs and MDSCs. Conclusions: The recognition of the microenvironment-myeloma cell interplay is essential for designing specific therapeutic strategies and ameliorating disease prognosis. Full article