The use of antibiotics to promote growth and prevent disease in livestock and poultry animals has raised concerns about antimicrobial resistance and its consequences for public health. Increasing pressure on the livestock and poultry industries demands improved productivity, reduced economic losses, and the assurance of food safety for human consumption. Plant bioactive compounds (PBCs) have emerged as safe and sustainable alternatives to antibiotics that promote the growth, support animal health and improve productivity without compromising food safety. PBCs represent a diverse group of secondary metabolites-including phenolics, terpenoids, polysaccharides, and organosulfur compounds-that exhibit a wide range of biological activities relevant to livestock and poultry nutrition and health. Recent studies have shown that supplementing animals with PBCs as feed additives, either as crude extracts or individual compounds, improves growth performance, nutrient utilization, and modulates gut microbiota, while also exerting antioxidant, anti-inflammatory, antimicrobial, and immunoregulatory effects, thereby enhancing overall resilience of animals. The multifunctional properties of PBCs, and the reduced risk of resistance development position them as promising candidates for next-generation feed additives for livestock and poultry production. However, challenges remain regarding optimal concentration, variability in bioavailability, stability during feed processing, and the standardization of active components. Here, we discuss the major PBCs and their potential as functional feed additives to improve livestock and poultry production. We further highlight research gaps and outline future prospects required to advance their adoption in sustainable animal production.
Foot-and-mouth disease (FMD) is among the most important transboundary animal diseases, causing widespread economic losses due to decreased productivity, trade restrictions, and costly disease management efforts. This study aimed to characterize circulating FMDV serotypes and quantify within-farm transmission dynamics during active outbreaks in Bishoftu, Ethiopia. Five epithelial tissue and 39 oral swab samples were collected from 11 dairy farms experiencing active FMD outbreaks. Samples were tested using quantitative reverse transcriptase polymerase chain reaction (RT-qPCR) and antigen-capturing enzyme-linked immunosorbent assay (ELISA) for FMDV detection and serotyping. A susceptible-infected-recovered (SIR) model was fitted to outbreak data to quantify transmission dynamics. Out of 44 samples, 37 (84.1%) were positive for FMDV using RT-qPCR, and 23 (52.3%) were positive by ELISA. Serotypes O, SAT-1, and SAT-2 were identified, with SAT-2 being predominant. Transmission modelling indicated an average transmission rate (λ) of 0.82 and a recovery rate (γ) of 0.32 per day, corresponding to a mean basic reproductive number (R0) of 3.7. The trivalent vaccine demonstrated low effectiveness (31% against single serotypes; 26% when adjusted for mixed infections), which was insufficient to achieve herd immunity given the high transmission intensity. These findings demonstrate that vaccination alone cannot control FMD in these systems. Control requires improved vaccine matching and the combination of vaccination with enhanced biosecurity measures, such as animal isolation and movement control.
Parkinson's disease (PD) is a progressive neurodegenerative disorder characterized not only by the gradual loss of motor control but also by a wide range of non-motor symptoms, including anxiety and depression, which can precede motor dysfunction by years. These psychiatric manifestations are commonly managed with selective serotonin reuptake inhibitors (SSRI). However, their impact on motor function in the early stages of PD remains controversial. In this study, we investigated the effects of fluoxetine in a progressive model of parkinsonism induced by repeated low-dose reserpine in male Wistar rats. Animals received short- (four injections) or middle-term (ten injections) reserpine (0.1 mg/kg, one every 48 h) treatment combined or not with daily fluoxetine (10 mg/kg) administration, and were evaluated for catalepsy, oral movements, open field behavior, body weight, and immunoreactivity for tyrosine hydroxylase (TH) and serotonin (5-HT). Fluoxetine decreased anxiety-like behavior, assessed by reducing the time animals spend exploring the center of the apparatus; potentiated reserpine-induced motor deficits, leading to earlier onset of cataleptic immobility, enhanced oral dyskinesia, reduced exploratory activity and body weight loss. In addition, fluoxetine alone decreased TH immunoreactivity in the dorsal striatum while increasing 5-HT labeling in the dorsal raphe nucleus, suggesting serotonergic modulation of dopaminergic circuits. In conclusion, fluoxetine administration prevented anxiety-like behavior but anticipated and exacerbated the motor changes caused by reserpine. Furthermore, simultaneous treatment with fluoxetine and reserpine worsened motor and behavioral impairments irrespective of TH changes, indicating that functional disturbances may precede dopaminergic degeneration. These findings highlight that fluoxetine, while targeting mood alterations, may exacerbate motor alterations during prodromal and initial stages of parkinsonism.
Many animals utilize pheromones as chemical substances that regulate reproductive and social behaviors. Elucidating how diverse pheromonal compounds are detected and translated into specific behavioral and physiological responses is crucial for understanding animal behavior. In vertebrates, pheromone reception is primarily mediated by the vomeronasal organ. Vomeronasal sensory neurons detect a broad range of semiochemical molecules including pheromones and allelochemicals, and transmit this information via the accessory olfactory bulb to higher brain regions, such as the amygdala and hypothalamic regions. Although recent studies have advanced the identification of pheromonal molecules and the analysis of the neural circuits responsible for their detection, ligands have been identified for only a small fraction of the hundreds of vomeronasal receptors. Analyses of genetically engineered mice lacking key components of vomeronasal signal transduction in vomeronasal sensory neurons have provided comprehensive insights into the relationship between vomeronasal sensory input and behavioral output. This review focuses on pheromones among the semiochemicals detected by the vomeronasal organ and summarizes current understanding of pheromonal perception and associated social behaviors, primarily based on studies in mice, the most extensively investigated model organism to date.
Fungal pathogens threaten the health of humans, animals, and plants. ITS sequencing offers an effective approach for detecting fungal pathogens; however, a comprehensive pathogen database and associated tailored pipeline are still lacking. This study introduces the multiple fungal pathogen detection (MFPD) pipeline, which incorporates an accurate and high-speed sequence alignment algorithm for broad-habitat pathogen identification. The curated MFPD database includes 95 660 full-length ITS sequences from 4924 reported fungal pathogen species. In silico experiments show that the full-length ITS achieves the highest accuracy in pathogen detection (average 99.34%), outperforming both the ITS1 and ITS2 subregions. Benchmarking against existing tools, including FUNGuild, FungalTraits, and ISHAM-ITS, shows that MFPD achieves the highest F1 scores in mock communities (0.89 for both plant and human-animal pathogens) and detects the broadest spectrum of pathogenic taxa in real samples. In addition to identifying causal pathogens, MFPD can also detect coinfecting pathogens in biological and environmental samples. Together, our work supports pathogen surveillance across diverse sectors, including clinical, agricultural, and livestock systems within a One Health framework.
L-Asparaginase (E.C.3.5.1.1; L-ASNase) hydrolyzes L-asparagine (an essential amino acid for the growth of leukemic cells) to aspartic acid and ammonia. It is obtained from various sources, including: bacteria, yeast, fungi, plants, and animals. It is used as a chemotherapeutic agent to treat acute lymphoblastic leukemia (ALL) and reduce acrylamide formation in baked and fried foods. Globally, various recombinant and pegylated L-ASNases formulations, including Escherichia coli and Erwinia-derived variants, are in Phase II or active clinical trials for ALL and related conditions across the USA, Brazil, and Canada. Some drugs are recruiting, and other variants of E. coli and E. chrysanthemi L-ASNases are already approved for marketing. Although L-ASNase is used as a therapeutic agent, immunogenic reactions and other adverse effects continue to limit its use. To enhance yield, L-ASNase isoforms are cloned and expressed in host cells, generating recombinants with distinct physicochemical properties and kinetic parameters. The enzymes' temperature and pH ranges vary from 25 to 100 °C and 6 to 10, respectively. Nowadays, enzymatic modifications, such as immobilization (chemical, physical, and PEGylation), mutagenesis, and PASylation, are used to overcome the limitations of commercialized L-ASNase-based drugs. Therefore, this review is a timely effort to compile and analyze the properties of recombinant L-ASNases and the contemporary techniques used to improve L-ASNase. A comprehensive study would help us better understand the kinetic parameters, biochemical properties, and modification trends of L-ASNase, enabling the development of robust, reliable therapeutics in the future. Currently available L-ASNase products often lack desirable pharmaceutical properties, including kinetic properties, increased half-life in blood serum, and decreased immunogenicity and toxicity. Several L-ASNases, including E. coli and Erwinia chrysanthemi, have been extensively characterized and evaluated in vitro for their anti-leukemic activity. However, only E. coli and Erwinia-derived formulations have advanced to preclinical animal models and subsequent clinical applications. Therefore, there is a critical need to identify new sources that are robust, more efficient, and have lower side effects. Hence, this review focuses on recombinant L-ASNases, their properties, drawbacks, and strategies for finding and improving L-ASNase variants.
Osteoarthritis (OA) is driven by a self-perpetuating cycle of cartilage wear, persistent inflammation, and lubrication dysfunction. Inspired by the natural superlubrication and piezoelectricity of healthy cartilage, we designed injectable hydrogel microspheres (HMBTLCs) that break this vicious cycle through a triple synergistic action of lubrication, anti-inflammation, and pro-regeneration. These microspheres are fabricated via a facile microfluidic strategy, integrating curcumin-loaded liposomes and barium titanate (BaTiO3, BT) nanoparticles within a single platform. The liposomes form a self-renewing boundary lubrication layer on the microsphere surface, achieving an ultralow coefficient of friction of 0.022, and provide sustained release of curcumin for over 28 days to scavenge inflammatory factors, thereby ameliorating the OA microenvironment. Concurrently, the embedded BT nanoparticles convert physiological joint loading into endogenous electrical signals (≈20 mV under 3 N load) to promote chondrogenesis. Critically, transcriptomic analysis reveals a cooperative mechanism: curcumin and piezoelectric stimulation not only synergistically upregulate chondrogenic markers (e.g., ACAN) and downregulate the catabolic enzyme MMP13 through distinct pathways, but also converge on shared anti-inflammatory signaling pathways (e.g., TNF-α, NF-κB, IL-17) to promote macrophage M2 polarization (CD206/CD86 ratio increased by 7.7-fold). By orchestrating these chemical and physical cues, HMBTLCs interrupt the pathological OA cycle under both cellular and animal conditions, reducing the OARSI score by 79.31% in a rat OA model, demonstrating their therapeutic potential for OA while also serving as a model for biomimetic material design.
In metazoan epithelia, arrays of motile cilia beat in a tissue-wide coordinated manner to drive unidirectional fluid flow. Whether radial spokes (RSs), which mediate mechano-signal transduction between the central pair and axonemal dynein arms, acquire metazoa-specific subunit(s) in evolution to facilitate this function remains poorly known. Here we report that Gα-GTPase activating protein family member RGS22 is a metazoa-specific component of RS1/2 complexes. Mouse Rgs22-deficient motile cilia exhibit ultrastructural defects, including deformation of the RS1 and disorganization of outer dynein arms. These abnormalities correlate with impaired planar polarity, manifested as unsynchronized ciliary beating and disrupted rotational polarity of basal bodies across the ependyma. The motility deficits further lead to progressive ciliary loss and shortening, resulting in postnatal hydrocephalus. Together, our data identify RGS22 an animal-lineage RS1/2 component required for normal RS architecture and associated with tissue-level ciliary motility and coordination phenotypes.
Are there differences in lifestyle and environmental exposures, and clinical characteristics between patients with histologically confirmed endometriosis residing in regional and metropolitan locations in Australia, and their impact on phenotype and disease severity? A retrospective study of patients with histologically confirmed endometriosis and detailed phenotypic mapping willing to participate in a Lifestyle and Environmental Risk Factor questionnaire. Demographic, lifestyle, environmental and clinical factors were compared between patients residing in regional and metropolitan locations in Australia, and between patients presenting with different lesion types. Body mass index (BMI; mean ± SD 27.16 ± 5.39 and 25.22 ± 4.93, respectively) and exposure to animal and plant toxins (14.0% and 3.0%, respectively; P = 0.0020) and pesticides (20.0% and 6.0%, respectively; P = 0.0059) were higher in regional participants compared with metropolitan participants. Increasing age (mean ± SD 36.26 ± 8.09, 40.84 ± 7.81 and 33.88 ± 8.39, respectively; P < 0.0001), leiomyomata (16.4%, 19.5 and 6.8%, respectively; P = 0.03) and infertility (63.9%, 63.4% and 40.9%, respectively; P = 0.04) were associated with higher rates of deep infiltrating endometriosis (DIE) and ovarian endometrioma (OMA) compared with superficial disease (SUP). Mental and behavioural problems were reported more often in patients with SUP and DIE than OMA (52.4%, 59.2% and 38.1%, respectively; P = 0.01). This study suggests significant differences in environmental exposures and lifestyle factors between patients with endometriosis living in regional areas of Australia compared with patients with endometriosis living in metropolitan areas of Australia. These findings emphasize the importance of environmental and lifestyle considerations in understanding the clinical variability of endometriosis.
Previous studies have demonstrated that Isthmin1 (Ism1) performs multiple biological functions and plays a role in immune responses. Nevertheless, the direct role of Ism1 in the context of bacterial infection remains poorly characterized. Here we characterized the anti-bacterial activity of Ism1 in zebrafish Danio rerio. The expression of zebrafish ism1 was found to be up-regulated upon the challenge with bacteria, as well as with lipoteichoic acid (LTA) and lipopolysaccharides (LPS), suggesting its involvement in the innate immune response. The recombinant Ism1 (rIsm1) could bind to both Gram-positive bacteria (Staphylococcus aureus, Bacillus subtilis) and Gram-negative bacteria (Escherichia coli, Aeromonas hydrophila), as well as the pathogen-associated molecular patterns (PAMPs) on the bacterial surfaces, including LTA, LPS and peptidoglycan (PGN), indicating that rIsm1 recognizes pathogens through these PAMPs. Furthermore, rIsm1 exhibited potent bactericidal activities against all tested bacteria. MIC/MBC assays showed that rIsm1 exhibited bactericidal activity against all four tested strains, with MIC values of 64 - 128 μg/ml and MBC/MIC ratios ≤4. Mechanistically, rIsm1 treatment was associated with disruption of cell membrane integrity, membrane depolarization, and increased intracellular reactive oxygen species (ROS) production. Moreover, in vivo bacterial clearance assays demonstrated that rIsm1 accelerated bacterial clearance in zebrafish embryos/larvae. Additionally, we observed extracellular localization of Ism1 in shrimp, zebrafish, and mice, suggesting a critical role in systemic immunity across different animal species. Collectively, our study demonstrates that zebrafish Ism1 serves as a key immunocompetent factor, defines its specific function and mode of action during immune responses, and provides new insights into the biological roles of Ism1 as well as the innate immunity in vertebrates.
Intestinal bacteria in healthy humans and animals are important reservoirs of antimicrobial resistance (AMR) genes. Certain Escherichia coli lineages dominate the global spread of AMR, but the extent of their host specificity and its impact on transmission remains unclear. Here, 1,198 E. coli isolates from cattle, chickens, humans, and pigs across Germany, the UK, Spain, and Vietnam were classified for host-association using a phylogeny-based bioinformatic approach, with non-host-associated isolates defined as 'generalists'. To validate this classification, 17 extended-spectrum beta-lactamase (ESBL)-producing isolates representing host-associated or generalist clusters were administered as a cocktail to three groups of weaner pigs, either untreated or treated with amoxicillin or ceftiofur. Bacterial shedding was monitored via culture and isolate-specific PCR up to 56 days post-inoculation. Colonization occurred within 24 h, with shedding peaking early and declining over time. Only 12 of 17 isolates were recovered post-inoculation; four were persistently detected, including one pig-associated, one bovine, one chicken, and one generalist isolate. These findings suggest limitations in the classification approach, possible low host selectivity in pigs, and/or incomplete knowledge of colonization mechanisms. Nonetheless, the model successfully identified isolates capable of stable colonization, supporting its utility for studying AMR E. coli host adaptation.
The (NRG1)-ERBB4 signaling pathway has been identified to be pathophysiologically meaningful for cognitive impairments in schizophrenia. The in the 1960s approved mineralocorticoid antagonist spironolactone has been shown to be effective to modulate (NRG1)-ERBB4 signaling in preclinical and animal models. Thus, a proof-of-concept drug repurposing trial in patients with schizophrenia was justified. We conducted a multi-site proof-of-concept randomized controlled trial. 90 patients with schizophrenia were randomized to a three-week trial of two different spironolactone dosages (100 mg or 200 mg) or placebo. The primary endpoint was predefined as change in working memory performance after three weeks of treatment. A naturalistic follow-up was performed nine weeks after the end of intervention. The trial was registered at the WHO International Clinical Trials Registry platform ( http://apps.who.int/trialsearch/Trial2.aspx?TrialID=EUCTR2014-001968-35-DE ). Despite large numerical improvements in working memory functions, particularly in the spironolactone 200 mg group, the pre-specified analyses do not demonstrate significant superiority of either intervention over placebo. However, post-hoc sensitivity analyses suggest a significant advantage of spironolactone for the primary endpoint. Safety measures show that both interventions are well tolerated. This finding suggests a potential positive effect of spironolactone on working memory in people with schizophrenia and the good safety outcomes may justify further trials with longer intervention periods or higher spironolactone dosages. Individuals diagnosed with schizophrenia frequently experience cognitive deficits, including challenges related to everyday memory and attention. We evaluated whether spironolactone, a drug available since the 1960s and today used to treat heart failure is effective in treating cognitive impairment in people with schizophrenia. For that, we conducted a placebo-controlled clinical trial where participants received either 100 mg or 200 mg spironolactone or placebo for three weeks. Before and after this intervention, we tested cognition. We were able to show that our intervention with spironolactone has the potential to improve cognition in people with schizophrenia. Further trials are needed before this intervention can be offered as part of the clinical routine.
The development of real-time imaging has progressed spectacularly in recent years, largely due to the availability of multiple fluorophores with distinct spectral properties. This progress enables the simultaneous imaging of an increasing number of targets, providing access to complex biological processes occurring during development, signaling, and metabolism. In this study, we successfully tested the transfer of three different fluorescence-activating and absorption-shifting tags (FAST), originally developed in animal systems and yeast, to plants. These small (14 kDa) proteins associate non-covalently and reversibly with specific chemicals known as fluorogens, generating bright fluorescence signals that can be reversibly eliminated by washing. This property offers versatile applications. Green- and Red-FAST proteins were successfully used to label proteins targeted to several plant cellular compartments. In contrast, the latest generation of FAST proteins, identified in Rheinheimera sp. A13L (RspA), which accepts multiple fluorogens and offers broad versatility in excitation and emission spectra, was used with a split FAST version to demonstrate cytosolic protein-protein interactions.
The common marmoset (Callithrix jacchus) is a neotropical primate widely distributed in Brazil and frequently associated with urban and peri-urban environments, a condition that favors exposure to different parasites. The present study aimed to report the occurrence of Primasubulura jacchi and to document the first record of Rhipicephalus sanguineus sensu lato parasitizing C. jacchus in Northeastern Brazil. A debilitated female C. jacchus was rescued in a peri-urban area of the municipality of Garanhuns, Pernambuco, and referred to the Veterinary Hospital of the Federal University of the Agreste of Pernambuco for clinical evaluation. Due to the severe clinical condition, the animal was euthanized. During necropsy, a fecal sample was collected directly from the rectum and analysed through egg flotation technique. Main macroscopic findings included extensive cutaneous lesions on the left thoracic and pelvic limbs, intense tick infestation and intestinal parasitism by nematodes. Ticks were morphologically identified as Rh. sanguineus s.l., whereas adult nematodes and eggs were identified as P. jacchi. This study expands the knowledge of the parasitic fauna associated with C. jacchus in Northeastern Brazil and reinforces the importance of health monitoring of primates in anthropized environments.
Despite advances in oncology, the dual challenge of achieving precise tumor-targeted therapy while simultaneously activating antitumor immunity remains a major clinical barrier. In this study, we engineered a biocompatible copper-based platform, hyaluronic acid (HA) modified Cu ions based therapeutic (B-Cu/HA), that integrates selective tumor targeting, intrinsic cytotoxicity, and immune activation within a single therapeutic system. Leveraging the HA-CD44 interaction, B-Cu/HA exhibited preferential accumulation and prolonged retention in CD44-overexpressing tumors, while maintaining an excellent biosafety profile. Across multiple cancer models, B-Cu/HA robustly inhibited tumor progression. Mechanistically, it induced cuproptosis through upregulation of FDX1 and aggregation of lipoylated DLAT, and triggered ROS-mediated activation of the cGAS-STING pathway, promoting immunogenic cell death. Transcriptomic analysis revealed activation of hypoxia and cytokine signaling pathways, aligning with enhanced CD8+ T-cell cytotoxicity and remodeling of the tumor immune microenvironment. The animal models studies demonstrated that B-Cu/HA significantly suppressed tumor growth without systemic toxicity, and synergistically enhanced the efficacy of the immune checkpoint inhibitors anti-TIGIT. Together, these findings establish B-Cu/HA as a multifunctional, immunomodulatory formulation that offers a clinically translatable strategy to enhance tumor immunotherapy and overcome resistance in CD44-overexpressing tumors.
Embryonic germ cells are the lineage restricted precursors to the gametes of the adult animal. Embryonic germ cells from birds offer a cellular system for both the conservation and the genetic modification of avian species. Ongoing discoveries in germ cell self-renewal pathways are extending the number of bird species for which germ cells can be in vitro cultured. Parallelly, the improved development of surrogate host embryos to carry exogenous germ cells is fostering more efficient and 3R-supportive methods to regenerate avian species from this cellular material. The next decade should lead to exciting developments which will aid in both understanding germ cell biology and avian conservation efforts.
Cisplatin is a common chemotherapeutic agent for advanced head and neck squamous cell carcinoma (HNSCC), but treatment success is often limited by resistance. Cancer stem cells (CSCs) are known contributors to this cisplatin chemoresistance in HNSCC. The mechanistic target of rapamycin (mTOR) pathway, which is frequently dysregulated in HNSCC, plays a crucial role via the PI3K/AKT/mTOR axis in maintaining CSC populations and promoting cancer proliferation. However, the specific effects of combining rapamycin, an mTOR pathway inhibitor, with chemotherapeutic agents on CSC maintenance and overall tumorigenicity remain unclear. We examined CSC gene expression in HNSCC cell lines (HSC4, SCC25, OT-1109) and evaluated the therapeutic potential of combining rapamycin, an mTOR pathway inhibitor, with cisplatin on CSC using a cell viability assay. The combination was further evaluated in an HSC4 mouse xenograft model. Tumor volume and animal weight were monitored throughout treatment. Xenograft tissue analysis via immunohistochemistry assessed stem cell markers (CD133 and ALDH1A1), proliferation markers (Ki-67), and mTOR pathway inhibition (pS6). Administration of low-dose cisplatin enriched the CD133+ cell population but failed to decrease the tumor mass in HNSCC xenografts. In contrast, the combination of cisplatin and rapamycin significantly impeded tumor growth and minimized toxicity, concurrently reducing the population of CD133+ tumor cells. These findings suggest that rapamycin enhances the mechanistic efficacy of cisplatin by specifically targeting and reducing cisplatin-induced stemness (CD133+ CSC population). This study proposes a viable combination therapy for HNSCC involving an mTOR inhibitor and a platinum-based drug to overcome CSC-mediated resistance.
Whole-genome sequencing (WGS) of Mycobacterium tuberculosis (Mtb) is widely used in epidemiological investigations of recent transmission events, resulting in high-resolution strain typing. Accurate and rapid strain typing is essential for informing outbreak investigations and guiding tuberculosis control strategies. However, the gold-standard reference-guided SNP-calling pipeline currently used for strain typing relies on computationally intensive reference-mapping approaches, making it challenging to perform in many high-burden, resource-limited settings, where simplified and scalable genomic tools are urgently needed. To address these limitations, we explored reference-free methods for medium-resolution epidemiology, namely, Mtb strain (lineage) typing, using a dataset of 535 complete genomes spanning the human- and animal-adapted lineages. Illumina paired-end reads were simulated from each complete genome, assembled and analysed using three reference-free, k-mer-based tools: MASH, PopPUNK and SKA2 (split k-mer analysis). Genetic distances were generated for each method and compared with a ground truth lineage assignment from TB-Profiler. Our results demonstrated that reference-free methods can effectively distinguish Mtb lineages, with SKA2 showing the most promising performance across all datasets. SKA2 consistently recovered lineage and sub-lineage structure with high accuracy, demonstrating strong potential as an alternative to traditional WGS workflows. These findings highlight the utility of reference-free methods, particularly SKA2, for enabling accessible, scalable and rapid Mtb strain typing, while supporting genomic epidemiology with low computational resources.
Hypoxia-inducible factor-prolyl hydroxylase (HIF-PH) inhibitors stimulate endogenous erythropoietin (EPO) production and are used for the treatment of anemia of chronic kidney disease (CKD) in humans. In veterinary medicine, molidustat (MOL) has demonstrated acceptable safety and efficacy with anemia of CKD in cats. Roxadustat (ROX) is another HIF-PH inhibitor that has a longer half-life than MOL, allowing for less frequent dosing. Therefore, ROX may offer an advantage in improving medication adherence. However, the safe dose range and erythropoietic effects of ROX in cats remain unclear. In the present study, an exploratory pharmacokinetic analysis of single-dose ROX administration was conducted in healthy cats to assess its safety and erythropoietic effects. In addition, a repeated-dose study was performed to evaluate safety and tolerability during continued administration. In the single-dose study, no apparent adverse events were observed at doses ≤20 mg/kg; however, severe adverse events including vomiting and systemic clinical signs leading to death were observed at 80 mg/kg, indicating that high-dose administration should be avoided. Following ROX administration, increases in reticulocyte counts and serum EPO concentrations were observed, suggesting erythropoietic activity in cats. In the repeated-dose study, ROX administered at 5-8 mg/kg for six weeks was not associated with apparent adverse events based on clinical condition and complete blood count. These results suggest that this dose range may serve as a candidate for future clinical evaluation in CKD cats.
Since the World Health Organization designated the Monkeypox (mpox) outbreak a Public Health Emergency of International Concern (PHEIC) in 2022, monkeypox virus (MPXV) has spread across all six continents. However, the virus's genomic variability and strong adaptability have thus far hindered the development of effective, targeted vaccines. In this study, bioinformatic analysis demonstrated that the A7 protein of MPXV is highly conserved among orthopoxviruses and contains hydrophilic B-cell epitopes. The recombinant A7 protein was expressed using a prokaryotic system, followed by purification and subsequent immunization of BALB/c and C57BL/6 mice. Immunization with A7 elicited robust humoral and cellular immune responses, accompanied by the induction of high-titer specific antibodies (with the highest titer reaching 1:409,600). The generated polyclonal antibodies exhibited high specificity and strong reactivity against eukaryotic-expressed A7 protein. These findings provide important evidence for understanding the immunogenicity of the MPXV A7 protein and lay a foundation for the development of mpox serodiagnostic reagents and further investigation into its potential as a subunit vaccine candidate.