Search Results (192)

Background: Colorectal cancer (CRC) is strongly influenced by miRNAs as well as the circadian system. Methods: High-throughput sequencing of miRNAs expressed in the rat colon during 24 h light (L)/dark (D) cycle was performed to identify rhythmically expressed miRNAs. The role of miR-150-5p in CRC progression was analyzed in DLD1 cell line and human CRC tissues. Results: Nearly 10% of mature miRNAs showed a daily rhythm in expression. A peak of miRNAs’ levels was in most cases observed during the first half of the D phase of the LD cycle. The highest amplitude was detected in expression of miR-150-5p and miR-142-3p. In the L phase of the LD cycle, the maximum in miR-30d-5p expression was detected. Gene ontology enrichment analysis revealed that genes interfering with miRNAs with peak expression during the D phase influence apoptosis, angiogenesis, the immune system, and EGF and TGF-beta signaling. Rhythm in miR-150-5p, miR-142-3p, and miR-30d-5p expression was confirmed by real-time PCR. Oncogenes bcl2 and myb and clock gene cry1 were identified as miR-150-5p targets. miR-150-5p administration promoted camptothecin-induced apoptosis. Expression of myb showed a rhythmic profile in DLD1 cells with inverted acrophase with respect to miR-150-5p. miR-150-5p was decreased in cancer compared to adjacent tissue in CRC patients. Decrease in miR-150-5p was age dependent. Older patients with lower expression of miR-150-5p and higher expression of cry1 showed worse survival in comparison with younger patients. Conclusions: miRNA signaling differs between the L and D phases of the LD cycle. miR-150-5p, targeting myb, bcl2, and cry1, can influence CRC progression in a phase-dependent manner. Full article

Serum cereblon (CRBN) has been proposed as a target protein for immunomodulatory drugs (IMiDs). IMiDs are one of the backbone treatment options in multiple myeloma (MM), rendering CRBN an intriguing candidate for use as a biomarker in clinical settings. Ninety-two (92) MM patients, mostly relapsed/refractory and a few at diagnosis, were included in the study, from lenalidomide–dexamethasone (LD) initiation until last follow-up or death. Median CRBN at LD initiation (N = 68) treatment was 247 pg/mL (range, 0–9760 pg/mL), at the time of best response (BR) status (N = 59) 142.5 pg/mL (range, 0–9940 pg/mL) and in patients with relapse/refractory MM to LD regimen (N = 54) 298 pg/mL (range, 0–9840 pg/mL). CRBN in healthy individuals was almost undetectable and significantly lower compared to the CRBN at LD initiation (p = 0.003), at BR to LD (p = 0.012) and at relapse to LD (p = 0.002). CRBN was significantly lower at BR in contrast to LD initiation and relapse to LD (p = 0.04, p = 0.028). High levels of CRBN at treatment initiation correlated with early relapse to LD (≤12 months) (p = 0.03). Seven-year survival was improved in patients with CRBN levels below median measured at the time of LD initiation (p = 0.013) as well as at BR (p = 0.032). CRBN was associated with treatment response and is predictive of survival after LD. Full article

Ralstonia eutropha H16, a metabolically versatile bacterium, has gained prominence as a microbial platform for sustainable bioproduction. While its capabilities in synthesizing single-cell proteins and biodegradable materials are well documented, comprehensive strain-level safety evaluations remain insufficient for food-grade applications. This study systematically assessed the safety of R. eutropha H16 through genomic, phenotypic, and toxicological analyzes. Genomic analyzes revealed the absence or minimal presence of virulence factors and antibiotic resistance genes, aligning with microbiological safety standards. Phenotypic investigations demonstrated a limited gastric fluid tolerance (pH 2.5, survival rate 25.70% after 3 h) and intestinal fluid persistence (pH 8, 44.67% viability after 3 h), coupled with an exceptional bile salt tolerance (0.2% w/v). Antioxidant assays confirmed the fermentation broth specifically scavenges DPPH free radicals (14.60 ± 1.24 μg Trolox/mL), whereas bacterial suspensions and cell-free supernatants exhibited a strong hydroxyl radical scavenging (>90 U/mL) and superoxide anion inhibition (>100 U/L). Acute toxicity testing indicated no mortality or histopathological abnormalities, with an LD₅₀ value exceeding 1 × 10¹¹ CFU/kg. Subacute toxicity studies (28-day, 1 × 10⁸–1 × 10¹⁰ CFU/kg) revealed no significant effects on growth, hematology, or organ function. Minor alterations in serum biochemistry might be attributed to physiological adaptation. Subacute exposure induced transient serum ALT fluctuations without hepatorenal dysfunction, while maintaining hematological parameters within physiological ranges. Collectively, these results substantiate the safety of R. eutropha H16 for food-related applications while underscoring the necessity of strain-specific risk assessments for industrial microbial platforms. Full article

The circadian rhythm of an animal’s body color change is crucial for its survival and adaptation to the environment, with photoperiod serving as a primary “zeitgeber” that significantly affects the circadian rhythm of color change. To explore the circadian body color change in juvenile Chinese giant salamanders (Andrias davidianus), four distinct photoperiod regimens were established, including the DD group (constant darkness), the LD12:12 group (12 h photophase, 12 h scotophase), the LD16:8 group (16 h photophase, 8 h scotophase), and the LD8:16 group (8 h photophase, 16 h scotophase). Reflectance spectra of dorsal skin were measured every 4 h over a 24 h cycle, with data collected every other day for three consecutive days, totaling 72 h. Reflectance spectra were converted into colorimetric variables to analyze circadian patterns. The results demonstrated that juvenile A. davidianus exhibited highly significant circadian rhythmicity under different photoperiods, with their body color becoming darker during the photophase (mean brightness: 14.650 ± 0.629 to 16.385 ± 1.301) and lighter during the scotophase (mean brightness: 16.473 ± 0.875 to 34.422 ± 2.692). Photoperiod alterations significantly affected the rhythm parameters (mesor, amplitude, and acrophase) of mean brightness variation. Compared with the LD12:12 group, the LD16:8 group (extended photophase) reduced rhythm mesor by 1.415, decreased amplitude by 0.988, and induced a 1.510 h acrophase delay. Conversely, the LD8:16 group (extended scotophase) increased rhythm mesor by 2.141 and amplitude by 2.919. These findings indicate that the circadian rhythm of body color change in juvenile A. davidianus is coordinately regulated by endogenous circadian rhythms and exogenous photoperiodic signals. The findings of this study provide a theoretical foundation for the conservation and management of A. davidianus. Full article

Our understanding of the molecular mechanisms undergirding artemisinin (ART) resistance in Plasmodium falciparum is currently based on two organizing principles: reduced hemoglobin trafficking into the digestive food vacuole, resulting in lower levels of activated ART, and increased tolerance to ART-induced oxidative stress in the infected erythrocyte. We had previously proposed an extracellular vesicle (EV) export model of ART resistance in P. falciparum. This model predicts that EV abundance will be altered by ART exposure and that the peptide cargo of EVs from the ART-exposed condition will be enriched with aggregation-prone peptides. We tested the predictions of the EV export hypothesis in this study using in vitro culture assays of an ART-resistant transgenic line engineered on a 3D7 background (R561H) and a 3D7 knock-out line (PfVps60KO) with deficient EV production phenotype. EV enrichment was obtained from in vitro parasite culture supernatants via a series of ultracentrifugation and filtration steps, followed by size exclusion chromatography. A quality check on EVs was performed using dynamic light scattering. Liquid chromatography with tandem mass spectrometry was used to determine the proteome cargo from extracted EVs, and parasite peptides were queried for aggregation-prone tendency using open-access software. We report that dihydroartemisinin (DHA) exposure was positively correlated with EV abundance (coefficient estimate = 1038.58, confidence interval of 194.86–1882.30, and p-value = 0.018) and suggests that EV biogenesis is part of the parasite’s response to DHA/ART. Furthermore, our findings suggest the expression of a non-constitutive DHA-induced alternate EV biogenesis pathway as the PfVps60KO was observed to produce the highest number of EVs under DHA exposure. Finally, we show that EVs from both ART-susceptible and resistant parasites under DHA exposure carry a cargo of Chorein N-terminal domain-containing protein (PF3D7_1021700) with a high aggregation-prone index (prion-like domain [PrLD] score = 26.5) out of nine identified parasite peptides. The former of these findings is in concordance with the EV export hypothesis, which posits that the removal of DHA/ART-induced aggregated and/or misfolded peptides is critical to the parasite’s survival under DHA/ART exposure. This observation further implicates EVs in the development of the ART-resistant phenotype. However, the finding of one aggregation-prone peptide out of the nine parasite proteins in the EV cargo does not sufficiently support the EV export hypothesis. Future replicates of this study and further interrogations of the EV export hypothesis are needed. Full article

The current work aims to establish an integrated pest management strategy using Hyperaspis trifurcata Schaeffer (Coleoptera: Coccinellidae) to control Dactylopius opuntiae Cockerell (Hemiptera: Dactylopiidae) and to assess the side effects of pesticides commonly used on this predator. The first part of this study was performed under controlled conditions at two temperatures with three prey densities and two release rates for 83 days. Under field conditions, a survival time test was conducted in a screen house (1.2 ha), where a total of 5700 predators were released on 1425 cactus plants and then monitored for a period of 23 weeks. Furthermore, eight pesticides were tested on H. trifurcata in laboratory conditions at five rates in order to define the lethal doses. Under controlled conditions, the effect of temperature on predation was not significant until 27 days after release. However, the prey density significantly impacted the predation rates from the 10th to 27th day after release (p < 0.001). The predator release rate significantly affected predation starting from the 15th day after release. The lowest median survival time based on Kaplan–Meier tests was obtained at 30 °C (the high temperature) for eight predators/cladode (27 days), but the highest was at 26 °C (the low temperature) for four predators/cladode (63 days). Depending on cochineal infestation, the effect of temperature significantly increased the predation rate from the 10th to 49th day after release, but only at a high density (50 colonies/cladode). Under field conditions, the effect of the infestation level on the survival function was significant (Log-Rank p < 0.05), and the median times were 111 and 130 days after release for low and high densities, respectively. Acetamiprid, Vaseline oil, black soap, copper oxychloride, and paraffin oil were highly toxic to H. trifurcata (>84% of mortality), and the LD₅₀ values ranged from 2.3 to 69.6% of the recommended rate. For Mancozeb at the recommended dose, the mortality rate was low (<2%). The large-scale release of H. trifurcata would be successful in the Near East and North Africa (NENA) region, provided that the use of the mentioned pesticides is avoided or at least reduced. Full article

The circadian clock, an intrinsic 24 h cellular timekeeping system, regulates fundamental biological processes, including tumor physiology and metabolism. Cancer stem cells (CSCs), a subpopulation of cancer cells with self-renewal and tumorigenic capacities, are implicated in tumor initiation, recurrence, and metastasis. Despite growing evidence for the circadian clock’s involvement in regulating CSC functions, its precise regulatory mechanisms remain largely unknown. Here, using a human osteosarcoma (OS) model (143B), we have shown that core molecular clock factors are critical for OS stem cell survival and behavior via direct modulation of CSC and lipid metabolic pathways. In single-cell-derived spheroid formation assays, 143B OS cells exhibited robust spheroid-forming capacity under 3D culture conditions. Furthermore, siRNA-mediated depletion of core clock components (i.e., BMAL1, CLOCK, CRY1/2, PER1/2)—essential positive and negative elements of the circadian clock feedback loop—significantly reduced spheroid formation in 143B CSCs isolated from in vivo OS xenografts. In contrast, knockdown of the secondary clock-stabilizing factor genes NR1D1 and NR1D2 had little effect. We also found that knockdown of BMAL1, CLOCK, or CRY1/2 markedly impaired the migration and invasion capacities of 143B CSCs. At the molecular level, silencing of BMAL1, CLOCK, or CRY1/2 distinctly altered the expression of genes associated with stem cell properties and the epithelial–mesenchymal transition (EMT) in 143B CSCs. In addition, disruption of BMAL1, CLOCK, or CRY1/2 expression significantly reduced lipid droplet formation by downregulating the expression of genes involved in lipogenesis (e.g., DGAT1, FASN, ACSL4, PKM2, CHKA, SREBP1), which are closely linked to CSC/EMT processes. Furthermore, transcriptomic analysis of human OS patient samples revealed that compared with other core clock genes, CRY1 was highly expressed in OS tumors relative to controls, and its expression exhibited strong positive correlations with patient prognosis, survival, and LD biogenesis gene expression. These findings highlight the critical role of the molecular circadian clock in regulating CSC properties and metabolism, underscoring the therapeutic potential of targeting the core clock machinery to enhance OS treatment outcomes. Full article

Background: Acute lung injury (ALI) is a major cause of death in patients with various viral pneumonias. Our team previously identified four volatile compounds from aromatic Chinese medicines. Based on molecular compatibility theory, we defined their combination as aromatic molecular compatibility (AC), though its therapeutic effects and underlying mechanisms remain unclear. Methods: This study used influenza A virus (IAV) A/PR/8/34 to construct cell and mouse models of ALI to explore AC’s protective effects against viral infection. The therapeutic effect of AC was verified by evaluating the antiviral efficacy in the mouse models, including improvements in their lung and colon inflammation, oxidative stress, and the suppression of the NLRP3 inflammasome. In addition, 16S rDNA and lipid metabolomics were used to analyze the potential therapeutic mechanisms of AC. Results: Our in vitro and in vivo studies demonstrated that AC increased the survival of the IAV-infected cells and mice, inhibited influenza virus replication and the expression of proinflammatory factors in the lung tissues, and ameliorated barrier damage in the colonic tissues. In addition, AC inhibited the expression of ROS and the NLRP3 inflammasome and improved the inflammatory cell infiltration into the lung tissues. Finally, AC effectively regulated intestinal flora disorders and lipid metabolism in the model mice, significantly reduced cholesterol and triglyceride expression, and thus reduced the abnormal accumulation of lipid droplets (LDs) after IAV infection. Conclusions: In this study, we demonstrated that AC could treat IAV-induced ALIs through multiple pathways, including antiviral and anti-inflammatory pathways and modulation of the intestinal flora and the accumulation of LDs. Full article

Background: A key component in modern vaccine development is the adjuvant, which enhances and/or modulates the antigen-specific immune response. In recent years, nanoparticle (NP)-based adjuvants have attracted much research attention owing to their ability to enhance vaccine potency. Nonetheless, how the selection of different antigens influences the overall vaccine efficacy when combined with the same nanoparticle adjuvant is less discussed, which is important for practical applications. Methods: Non-toxic mutants of exotoxin Hla (rHlaH35L) and cell-wall-anchored protein SpA(rSpam) were covalently conjugated to Poly(lactic-co-glycolic acid)-polyethylene glycol (PLGA-PEG) 25% NPs (25% NPs) as antigens to prepare nanovaccines. Antibody titers, cytokine secretion levels, and the antibody bacteriolytic capacity were tested to investigate immune activation. To evaluate the protective efficacy of the nanovaccine, immunized mice were challenged with S. aureus ATCC 25923 at three different lethal doses: 1 × LD₁₀₀, 2 × LD₁₀₀, and 4 × LD₁₀₀. Results: We showed that 25% NP-rHlaH35L nanovaccines were associated with more efficient humoral, cellular, and innate immune responses and protection potency compared with 25% NP-rSpam. Moreover, the overall vaccine potency of 25% NP-rHlaH35L was even better than the combination vaccination of both 25% NP-rHlaH35L and 25% NP-rSpam. In comparison to the clinically used aluminum (alum) adjuvant, the 25% NP adjuvants were found to stimulate humoral and cellular immune responses efficiently, irrespective of the antigen type. For antigens, either exotoxins or cell-wall-anchored proteins, the 25% NP-based vaccines show excellent protection for mice from S. aureus infection with survival rates of 100% after lethal challenge, which is significantly superior to the clinically used alum adjuvant. Moreover, due to the superior immune response elicited by 25% NP-rHlaH35L, the animals inoculated with this formulation survived even after two times the lethal dose of S. aureus administration. Conclusions: We demonstrated that the type of antigen plays a key role in determining the overall vaccine efficacy in the immune system when different kinds of antigens are conjugated with a specific nanoparticle adjuvant, paving a new way for vaccine design based on 25% NP adjuvants with enhanced potency and reduced side effects. Full article

Botulinum neurotoxins (BoNTs) are the most potent toxins on Earth and are classified as Category A biological agents. BoNTs lead to paralysis in humans and cause botulism. Antibody therapeutics can effectively treat toxin-mediated infectious diseases. In this study, we generated a pharmaceutical humanized monoclonal antibody (HZ45 mAb) to prevent or treat botulism. HZ45 binds to the heavy chain receptor (HCR) domain of the toxin, preventing the toxin from entering the cell. The mAb was produced using hybridoma technology and phage display. We evaluated HZ45 mAb for the neutralization of BoNT serotype A (BoNT/A) in mice and rabbits. The survival results showed that pretreatment with HZ45 mAb provided 100% protection at a dose of 0.1 mg per mouse against a maximum of 100 LD₅₀ of BoNT/A. To assess the therapeutic efficacy of HZ45 mAb in New Zealand white rabbits (NZWs), a 5 mg dose was administered 4 or 8 h after challenge with 10 LD₅₀. The results indicated that 5 mg of HZ45 could treat the NZWs within 8 h after exposure to 10 LD₅₀ botulinum. Consequently, in an in vivo context, including mice and rabbits, HZ45 mAb could protect against botulinum type A intoxication. Full article

Aedes aegypti mosquitoes are established throughout Nouakchott, Mauritania, where its insecticide resistance status is unknown and dengue has become endemo-epidemic since 2014. Eggs were collected using ovitraps at 12 sites in five districts of Nouakchott, in August 2024. Adult females and larvae of the F1 generation were used for bioassays. Permethrin, deltamethrin, bendiocarb, and malathion were evaluated at discriminating concentrations. Larval assays were carried out at seven concentrations with Bacillus thuriengensis var israelensis (Bti) and temephos. The presence of knockdown resistance (kdr) mutations known to be associated with pyrethroid resistance was assessed by polymerase chain reaction and amplicons sequencing. Adults showed high levels of resistance to all insecticides tested. Larvae were susceptible to Bti (LD50 < 50 µg/L) and temephos (LD50% = 6.8 ± 0.7 µg/L). Only three kdr point mutations, S989P, V1016G, and F1534C, were found. The tri-locus genotypes SP/VG/FC were significantly associated with pyrethroid survival while only the tri-locus genotypes PP/GG/FF showed significant association with deltamethrin resistance. Given their level of insecticide resistance, there is an urgent need to control Ae. aegypti populations by several methods, including the use of biological larvicides, physical elimination of peridomestic breeding sites, water drainage, and public education to prevent arbovirus transmission. Full article

An experimental ecological method was used to study the effects of water temperature, photoperiod, and light intensity on the survival, feeding, and growth of juvenile Schizothorax irregularis and Diptychus maculates. The Box–Benhnken experiment was designed to predict the optimal environmental conditions for juvenile growth. With the maximum specific growth rate at 15 °C and a photoperiod of LD16:8, the results demonstrated that the juvenile S. irregularis had a survival rate of over 85% in water temperatures ranging from 5 to 25 °C. A daily light duration of 15.86 h and a light intensity of 1166.28 lx, with the water temperature maintained at 10.45 °C, allowed the juvenile S. irregularis fish to attain the optimal circumstances for growth and survival. At water temperatures below 25 °C, the juvenile D. maculates exhibited maximum specific growth rates at 10 °C and LD16:8 light period. Additionally, as the light intensity reached 1000 lx, the juvenile fish grew better. Furthermore, the juvenile D. maculates fish achieved theoretically optimal survival and growth circumstances when the water temperature was maintained at 10.87 °C with a light period of 15.0.5 h per day and a light intensity of 1474.68x. The results showed that both fish species may be raised in captivity in highland regions, but precise control over water temperature is required. Full article

The essential oil (EO) of Schinus areira exhibits a chemical composition dominated by monoterpene and sesquiterpene hydrocarbons, with α-phellandrene, limonene, α-pinene, and p-cymene as major constituents. This study aimed to evaluate the effects of S. areira EO on the biology and behavior of the Mediterranean fruit fly, Ceratitis capitata, particularly its attraction to the EO and the impact on its reproductive behavior and survival. Females were attracted at the initial choice and the time spent in the arm of the Y-tube olfactometer with the EO was longer, while males were attracted at the final choice, indicating the attractive potential of S. areira EO for both sexes of C. capitata. Within the context of the sterile insect technique (SIT), the better performance of released sterile males allows more copulations with wild females in competition with wild males, increasing the efficacy of the SIT. Exposure of tsl sterile males to the EO did not enhance their sexual competitiveness and increased latency to initiate copulation, indicating potential adverse effects. In addition, in oviposition assays, only a low concentration of the EO stimulated egg-laying on treated substrates, possibly due to the absence of deterrent compounds such as linalool. Finally, the LD50 of the EO was <25 µg/fly for both females and males, at 72 h post-treatment. These findings highlight the potential of EOs as biopesticides that influence the behaviors of C. capitata and emphasize the need for further studies to optimize their application in integrated pest management strategies, including the SIT. Full article

DNA methylation is a critical mechanism for regulating gene expression in bacteria and plays an essential role in bacterial pathogenesis. A mutant, WC1535ΔhsdM, lacking hsdM encoding a DNA methyltransferase was constructed using homologous recombination technology. The growth, hemolytic activity, and capsule formation of the mutant were analyzed. The dynamic distribution of the wild-type (WT) and mutant strains in tilapia tissues after artificial infection was determined. The adhesion, invasion, anti-phagocytic, and whole-blood survival abilities of the WT and mutant strains were analyzed. Tilapia were intraperitoneally injected with the WT or mutant strains, and the LD₅₀ values were determined. The expression levels of the immune-related genes in tilapia were analyzed by qRT-PCR. The mutant showed faster growth during the logarithmic growth period (5–10 h) and lower hemolytic activity than the WT strain. Mutant loads in tilapia tissues were significantly lower than those of the WT strain. Mutant strain adhesion to epithelial cells was significantly reduced, it was more easily engulfed by macrophages, and it had decreased intracellular survival. The LD₅₀ of the mutant was 2.06 times higher than that of the WT strain, indicating decreased pathogenicity. Expression levels of immune-related genes IL-1β, IL-6, IFN-γ, and TNF-α in tilapia induced by the mutant were lower than those by the WT strain. In conclusion, the WC1535ΔhsdM mutant exhibited an increased growth rate and decreased hemolytic activity, tissue colonization, and pathogenicity, indicating that HsdM could regulate S. agalactiae growth and pathogenicity. This study provides new insights into the pathogenesis of piscine S. agalactiae. Full article

Photobacterium damselae subsp. Piscicida (PDP), a marine bacterium, has been reported to infect a variety of economically important marine species worldwide. Understanding the occurrence and pathogenicity of PDP is crucial for effective disease control and ensuring the success of aquaculture operations. In late August 2023, an epidemic outbreak of P. damselae subsp. piscicida DQ-SS1, accompanied by significant mortality, was recorded in cage-cultured black rockfish (Sebastes schlegelii) located on Daqin Island for the first time. Genomic analysis revealed that DQ-SS1 possesses 2 chromosomes, with a total size of 4,510,445 bp and 3923 predicted CDSs. Pathogenic genes analysis identified 573 and 314 genes related to pathogen–host interactions and virulence, respectively. Additionally, DQ-SS1 displayed susceptibility to 15 antimicrobials, was resistant to 11 antimicrobials, and was intermediately sensitive to four antibiotics. Meanwhile, the in vitro assay revealed that the extracellular products (ECP) of DQ-SS1 were lethal to macrophages and exhibited hemolysin, lipase, and amylase activities. Moreover, DQ-SS1 also demonstrated the ability to survive in fish serum and resist complement-mediated killing. The in vivo assay showed that the infected fish exhibited severe histopathological alterations, such as the infiltration of inflammatory cells, cellular degeneration and necrosis, and loose cell aggregation. Lastly, the in vivo infection assays revealed the LD₅₀ of DQ-SS1 was 1.7 × 10³ CFU/g. This is the first study to elucidate the pathogenicity and genomic characteristics of multidrug-resistant PDP in cage-cultured S. schlegelii, which contributes to the advancement of diagnostic and preventative strategies for this disease in marine-cultured fishes and provides information for an in-depth study of the pathogenic mechanism of PDP. Full article