Search Results (618)

The Zinc Finger X-Linked Duplicate B (ZXDB) gene is one of a pair of replicated zinc finger genes on chromosome Xp11.21. The homologous gene of ZXDB in mice is Zxdb. Recent studies have found that Zxdb plays a role in the spermatogenic process of mice; however, its impact on the female reproductive system has not yet been explored. In our study, we found, for the first time, that the loss of function of Zxdb leads to reduced decidualization rates and a decrease in litter size in female mice. Secondly, we found that maternal loss of Zxdb is the determinant of these phenotypes. Thirdly, the transcriptional and proteomic differential expression genes in the uterine tissues of wild-type (WT) and Zxdb knockout (Zxdb-KO) mice were significantly enriched in signaling pathways such as adhesion molecules. Finally, we demonstrated that the disorder of expression and uneven distribution of adhesion molecules in mouse uterine tissue may be the main reason for the decline in embryo implantation rate. In conclusion, we have established for the first time a link between the Zxdb gene and reduced female fertility. This study will help provide guidance and genetic counseling for future common clinical complications such as Recurrent Spontaneous Abortion (RSA) or Recurrent Implantation Failure (RIF). Full article

Molecular sex determination in Syrian hamsters (Mesocricetus auratus) has been limited by the incomplete annotation of Y-linked loci in currently available genome assemblies. Here, we evaluate the Y-linked gene PRSSLY, which encodes a testis-specific serine protease-like protein, as a molecular marker for genetic sexing of Syrian hamster embryonic and placental tissues. Primers flanking a conserved PRSSLY coding region produced a male-specific amplicon showing 100% concordance with results from the established KDM5C/KDM5D PCR assay in E15.5 tail biopsies. SYBR Green–based qPCR enables the accurate detection of PRSSLY, characterized by a unique melt-curve profile, exclusively in male samples, allowing for efficient and sensitive mid-throughput analysis. Application of the PRSSLY assay to 417 placental samples from 39 dams demonstrated its suitability for large-scale sex genotyping, enabling sex assignment in the majority of samples despite the intrinsic complexity of placental tissue containing both maternal and embryonic genetic material. This assay provides a robust and reproducible approach for accurate sex genotyping in developmental and reproductive studies using Syrian hamsters. Full article

Canine parvovirus type 2 (CPV-2) is a primary etiological agent of acute gastroenteritis in domestic dogs. Although molecular and serological evidence have confirmed its circulation in wild carnivores, the clinical impact of spillover events in wildlife hosts remain insufficiently characterized. In this study, we investigated CPV-2 from wild coyote pups (Canis latrans) presenting with clinical gastroenteritis in northeastern Mexico. CPV-2 was successfully isolated in MDCK cells, and whole-genome sequencing was performed on two isolates, B55 and B56 (GenBank accession numbers PQ065988 and PQ065989). A comprehensive analysis identified 23 nucleotide mutations, eight of which were missense mutations resulting in amino acid substitutions in structural (VP) and non-structural (NS) proteins. Notably, amino acid substitution L354V was identified in the NS1 helicase domain of both isolates, a region critical for viral replication. Phylogenetic analysis confirmed that isolates B55 and B56 cluster within the CPV-2c subtype, showing high genetic relatedness to circulating Mexican and US canine strains which strongly suggests recent cross-species transmission between domestic dogs and wild coyotes. This study provides the first complete genomic characterization of a clinical CPV-2 infection in wild coyotes in Mexico, underscoring the immediate risk of CPV-2c transmission at the domestic animal–wildlife interface. Full article

Necrotizing enterocolitis (NEC) is a life-threatening inflammatory disease of preterm infants, increasingly viewed as a cytokine-driven disorder of the immature intestine. We aimed to characterize local peritoneal concentrations of interleukin (IL)-1β, IL-6 and tumor necrosis factor-α (TNF-α) in a standardized neonatal rat NEC model and relate them to histopathological injury. Seventy-four SPRD/Mol/Lodz rat pups were allocated to a control group (CTRL; n = 12) or subjected to a hypoxia-hypothermia-formula-feeding NEC protocol (NEC; n = 62). After 72 h, small-intestinal samples were scored using a four-tier NEC scale (0–3), and peritoneal fluid cytokine levels were measured by ELISA. All CTRL animals exhibited normal histology (grade 0), whereas NEC pups showed a wide spectrum of lesions, with 66.6% classified as grade 2–3 and a significantly higher mean NEC score in NEC than CTRL (p < 0.001). Peritoneal IL-1β and TNF-α concentrations were markedly elevated in NEC versus CTRL animals (both p < 0.001), while IL-6 levels showed no statistically significant between-group difference. These findings indicate that experimental NEC in this model is accompanied by a pronounced local pro-inflammatory response dominated by IL-1β and TNF-α, whereas IL-6 may follow distinct temporal or compartment-specific kinetics. Peritoneal cytokine profiling may help refine mechanistic understanding and guide future biomarker and immunomodulatory strategies in NEC. Full article

New Zealand fur seals (Arctocephalus forsteri) were severely exploited by historical hunting. However, recently assessed colonies in New Zealand are mostly thought to be growing or stable. The exceptions are three colonies (Wekakura Point, Cape Foulwind and Taumaka Island) on the West Coast of the South Island (‘WCSI’), previously documented as in decline. We used mark-recapture and morphometric data to update understandings of pup abundance and condition at these colonies. Pup abundance has continued to decline. In 2025, 186 (95% CI = 178–194) pups were estimated at Wekakura Point, 131 (95% CI = 122–140) at Cape Foulwind and 566 (95% CI = 555–577) at Taumaka Island, representing declines of 83%, 71% and 61% from the respective maxima in the 1990s. Rates of decline have slowed at Wekakura Point and Cape Foulwind since 2016 but have increased at Taumaka Island. Pup condition demonstrated substantial interannual variation. Cape Foulwind pups had the greatest average mass and body condition index score, followed by Wekakura Point and then Taumaka Island. There have been consistencies between years of particularly low pup abundance and condition across the colonies, suggesting common stressors; however, there are likely also some localised factors. Emerging diseases and marine environmental change are evaluated as potential drivers. Full article

β-casein (β-CN) is the predominant casein fraction in breast milk, while current infant milk formula (IMF) contains substantially lower β-CN levels than breast milk. The impact of β-CN fortification on neonatal digestive characteristic and bioactive peptide release remains an understudied area in vivo. This study investigated the effect of β-CN fortification in milk protein on digestion properties and release of bioactive peptides using a suckling rat pup model. Rat pups were, respectively, gavaged with two milk protein solutions: one with ordinary β-CN content (OBCN) and the other with fortified β-CN content (FBCN). The gastric emptying rate, proteolytic efficiency, and peptidomic profiles of intestinal digesta were evaluated. Results indicated that the FBCN group exhibited accelerated gastric emptying into the intestinal phase and enhanced proteolytic efficiency compared to OBCN group. Furthermore, the FBCN group generated greater peptide diversity in the small intestine, with significantly elevated abundance of bioactive peptide candidates exhibiting broader functional spectra. These findings provide additional evidence for the health effects of β-CN fortification in IMF. Full article

Background/Objectives: Preterm infants are at risk of developing the chronic lung condition of bronchopulmonary dysplasia (BPD), with associated alveolar simplification and airway hyperreactivity. Inhibition of S-nitrosoglutathione (GSNO) reductase has been shown to rescue airway hyperreactivity in a murine model of BPD. Here, we investigate the effects of early treatment with N6022, a pharmacologic GSNO reductase inhibitor. Methods: Newborn C57BL/6 mice were exposed to either 21% (control) or 60% oxygen (BPD model) for 5 days after birth. Pups simultaneously received either subcutaneous saline or varying doses of N6022 for 5 days during hyperoxia exposure. Pups were then recovered in room air to 3 weeks postnatal age. H&E-stained lungs were analyzed for alveolar simplification and airway tethering. In vivo airway reactivity to inhaled methacholine was measured using a flexiVent system. In separate littermates, lungs were immediately harvested after 5 days of hyperoxia for protein quantification via automated capillary Westerns. Results: Alveolar simplification and decreased airway tethering were noted in the 60% + saline group. Pups treated with N6022 during hyperoxia displayed dose-dependent improvements in alveolar simplification and airway tethering. Similarly, hyperoxia-exposed pups had increased airway reactivity, as measured by elevated respiratory system resistance and elastance responses to methacholine. Treatment with 10 mg/kg/day N6022 during hyperoxia resulted in decreased resistance and elastance responses. TGF-β expressions were elevated in the 60% + saline group and attenuated in the 60% + N6022 groups. Conclusions: Early exposure to GSNO reductase inhibitors such as N6022 can prevent hyperoxia-induced alveolar simplification and airway hyperreactivity, with lasting effects even after cessation of treatment. Full article

Flowers are key reproductive structures for many plant species. They are essential for seed and fruit production, and their development is tightly regulated by hormonal and genetic networks. The homeodomain transcription factors HAT3 and ATHB4 are known regulators of adaxial identity and hormone response. We demonstrate that flowers of the hat3 athb4 double mutant emerge at wider divergence angles relative to the wild type, a phenotype reflecting modified phyllotaxy and regulated by low auxin conditions. In addition, hat3 athb4 flowers exhibit aberrant trichome patterning on their sepals associated with enhanced sensitivity to cytokinin (CK). Through RNA-seq analysis of hat3 athb4 inflorescences, we identify the misregulation of genes involved in auxin biosynthesis (YUCCAs), auxin transport (PID), and CK metabolism (CKXs) and transport (PUPs). These findings suggest that HAT3 and ATHB4 fine-tune the auxin/CK balance and coordinate critical pattern events during reproductive development, offering new insight into hormone-mediated regulation of floral patterning. Full article

The reproductive biology of the whale shark (Rhincodon typus), the world’s largest fish, remains poorly understood, in large part due to the rarity of observations of neonates and of breeding behaviours. Although several regions in Indonesia, including Saleh Bay (West Nusa Tenggara Province), have been identified as aggregation and sighting sites for juvenile whale sharks (2–7 m total length, TL), smaller individuals from these potential nursery areas have not been previously documented. In August 2024, fishermen operating lift-net fishing vessels (bagans) in eastern Saleh Bay reported five separate sightings of a small whale shark estimated at 1.2–1.5 m TL and approximately four months old. Subsequently, on 6 September 2024, a male neonate measuring approximately 135–145 cm TL, estimated to be around four months old, was incidentally caught inside a bagan lift-net. These observations represent the first records of neonatal whale sharks in Indonesia and among the smallest free-swimming individuals ever documented globally, and suggest that Saleh Bay may serve as a pupping and early nursery area for whale sharks. These findings highlight the ecological significance of Saleh Bay for the early life stages of whale sharks and underscore the importance of collaborative monitoring and citizen science involving bagan fishermen in advancing the research and conservation of this endangered species. Full article

(1) Background: How variations of the push-up plus (PUP)—particularly changes in the base of support and scapular excursion—affect scapular muscle activation remains unclear. This study compared phase-specific electromyographic (EMG) activity of scapular muscles during four protraction–retraction exercises. (2) Methods: Twenty-six healthy male participants (age: 22.88 ± 1.45 years; height: 1.74 ± 0.05 m; weight: 77.31 ± 8.61 kg; body mass index (BMI): 25.61 ± 2.43 kg/m²) with Pilates experience performed four scapular protraction–retraction exercises under two base-of-support (quadruped vs. single-leg) and two movement-range (PUP vs. STD) conditions. Exercise order was randomized, and sufficient rest intervals were provided to minimize fatigue effects. Surface electromyography was recorded from six scapular muscles and normalized to maximal voluntary isometric contraction. The study was registered on CRIS (KCT0010032). (3) Results: Single-leg PUP showed the greatest serratus anterior (SA) activation, with increases of approximately 30% in protraction, 20–25% in isometric, and 15–20% in retraction. STD variations elicited higher trapezius activation, especially during large scapular excursions. The UT/SA ratio was significantly lower in PUP conditions (η²_p = 0.544), reflecting a more favorable stabilization pattern. (4) Conclusions: This experimental repeated-measures study demonstrates that combining single-leg support with traditional PUP meaningfully increases SA recruitment across all phases, whereas increased scapular range enhances trapezius engagement. These findings provide novel phase-specific insights into how PUP variations modulate closed-chain scapular stabilization and may assist clinicians in selecting targeted exercises. Interpretation should be limited to trained healthy males. Full article

Background/Objectives: Breast milk lysozyme is crucial for infant intestinal health. The low breastfeeding rate has driven the investigation of alternatives like hen egg white lysozyme (HEWL) for infant formula supplementation. However, HEWL differs significantly from human lysozyme. This study aimed to systematically compare the functional efficacy of recombinant human lysozyme (rhLYZ) and HEWL to assess their suitability as formula supplements. Methods: The physicochemical properties (enzymatic activity, optimal pH, thermal stability) of rhLYZ and HEWL were analyzed. Biological functions were evaluated using HT-29 intestinal cells for proliferation, differentiation, and protection against lipopolysaccharide-induced damage. In vivo effects on growth, intestinal morphology, and gene expression were assessed in a mouse pup model via transcriptomic analysis. Gut microbiota composition was also examined. Results: rhLYZ exhibited twice the enzymatic activity of HEWL, with an optimal pH of 6.0. In cellular models, rhLYZ enhanced intestinal epithelial differentiation at low concentrations. In vivo, rhLYZ supplementation significantly improved pup body weight, intestinal maturity, and villus-to-crypt ratios, outperforming HEWL. Transcriptomics revealed rhLYZ upregulated broad-spectrum antimicrobial peptides (e.g., Defa, lactoferrin) and immune-related genes, whereas HEWL induced a narrower antibacterial response and downregulated key defensins. Furthermore, rhLYZ significantly increased gut microbiota diversity and enriched beneficial butyrate-producing bacteria. Conclusions: rhLYZ more effectively mimics human milk lysozyme by promoting intestinal development, broad-spectrum immunity, and a balanced microbiota. HEWL shows a narrower functional profile. These findings provide a scientific basis for optimizing lysozyme selection in infant formula, highlighting the superior potential of rhLYZ. Full article

The assessment of animal welfare in rehabilitation settings is a critical aspect of effective care, yet typical metrics often fail to fully capture rehabilitating animals’ emotional experiences in a non-invasive way. Anticipatory behavior has emerged as a promising animal welfare indicator, reflecting an animal’s perceived need for rewards based on available opportunities in their environment. By tracking anticipatory responses, caretakers can gain insight into an animal’s reward sensitivity and use this information to guide management interventions. This study investigated the effects of enrichment type on anticipatory behavior in fourteen, rehabilitating harbor seal pups (Phoca vitulina richardii). We provided pups with daily sessions of either structural or cognitive enrichment and recorded their behavioral responses. During scheduled feeding sessions, we identified behaviors that emerged as anticipatory, then measured the frequency and duration of anticipatory behavior prior to the feeds to assess how enrichment types influenced the seals’ reward sensitivity, and thus their welfare. While enrichment interaction did not directly modulate anticipatory behavior, we observed a trend suggesting that exposure to cognitive enrichment reduced anticipatory behavior duration compared to structural enrichment. These findings align with previous research in zoo settings, where cognitive enrichment has been linked to improved welfare through reduced anticipatory behavior, though this effect has not been explored in a wildlife rehabilitation context. This study highlights the value of anticipatory behavior as a practical welfare assessment tool in rehabilitation settings and underscores the potential for enrichment, particularly cognitive, to improve welfare in rehabilitating marine mammals. Full article

Alcohol (ethanol; EtOH) intake affects one in ten pregnancies in the United States and is a leading cause of developmental defects collectively known as fetal alcohol spectrum disorders (FASDs). Cerebral circulation is a critical target of prenatal ethanol exposure (PEE), yet the target(s) involved remain poorly understood. In adult cerebral circulation, mitochondrial function is essential in regulating smooth muscle contractility, suggesting mitochondria as a potential target of alcohol in the developing cerebral arteries. In this study, pregnant C57BL/6J mice were administered ethanol (3, 4.5, 6, or 7 g/kg) during either the second trimester equivalent of human pregnancy (gestational days 9–19), or the third trimester equivalent during postnatal days 1–10. Maternal and progeny blood ethanol concentrations, progeny brain weight, cerebral artery oxygen consumption, and corticosterone levels were measured. At lower ethanol concentrations (3 g and 4.5 g/kg), no significant alterations in fetal cerebral artery mitochondrial function were detected. In contrast, heavy maternal ethanol exposure (6 g/kg) significantly increased mitochondrial respiratory parameters in developing cerebral arteries during the third trimester equivalent of human pregnancy. Sex-specific dimorphism was also observed at this developmental stage. Corticosterone was not elevated in fetuses and pups. In summary, our findings demonstrate developmental stage- and sex-dependent vulnerabilities of cerebrovascular oxygen consumption to ethanol exposure. Full article

Aquaporin-4 (AQP4) is expressed in ependymal cells bordering the ventricles, the glia limitans, and pericapillary astrocyte endfeet forming the blood–brain barrier. The sporadic occurrence of obstructive congenital hydrocephalus (OH) has been observed in the offspring of AQP4⁻/⁻ mice generated in the CD1 strain background. Here, we used microarray analysis to explore gene expression profiles in the periaqueductal area from littermate AQP4⁻/⁻ pups at postnatal day 12. We compared wild-type (WT) animals with AQP4⁻/⁻ animals that developed OH (AQP4⁻/⁻-OH) and those that did not (AQP4⁻/⁻-NH). Bioinformatic analysis identified gene sets associated with proliferation and migration of microglia, ependymal cell adhesion, extracellular matrix components, axon myelination, and neuronal synapsis. Among the differentially expressed genes, Spp1—expressed by neonatal CD11c⁺ microglia—was highlighted in the triple comparison. Spp1 was significantly upregulated in AQP4⁻/⁻-NH and downregulated in AQP4⁻/⁻-OH mice. These findings suggest that CD11c⁺ microglia, via Spp1 expression, play a key morphogenic role in the aqueduct of Sylvius and their absence, occurring in a small subset of AQP4⁻/⁻-CD1 animals, leads to obstructive hydrocephalus. Full article

Background/Objectives: Early life is crucial for infant gut development and intestinal homeostasis. Lactoferrin (LF) and osteopontin (OPN) are bioactive breast milk proteins that are supplemented into infant formula to promote gut development. However, the combined effect of LF and OPN (LOP) on in vivo gut maturation has not been fully elucidated. This study investigated the effects of LF, OPN, and LOP on intestinal epithelium maturation in C57BL/6N mice from postnatal days 7 to 21. Methods: 3-day-old pups were assigned to four groups: Control group, LF group: 300 mg/kg LF; OPN group: 300 mg/kg OPN, LOP group: 300 mg/kg of a 1:5 (w/w) mixture of LF and OPN. Results: Compared to controls, LOP reduced plasma Diamine Oxidase (DAO) activity by 1.54-fold and D-lactate levels by 1.41-fold, demonstrating greater efficacy than LF or OPN alone in reducing intestinal permeability. LOP also significantly increased intestinal absorptive cells versus controls or single proteins. Mechanistically, LOP promoted directional intestinal stem cell differentiation, increasing jejunal transit-amplifying cells by 1.40-fold in 21-day-old mice. LOP upregulated expression of the Notch pathway target Hes1 by 1.70-fold. Further investigations revealed LOP activated Notch signaling via the transcription factor Brg1. Validation using intestinal organoids and IEC-6 cells confirmed intact OPN within LOP mediates increased Brg1 expression, activating the Notch pathway to direct intestinal stem cell differentiation into absorptive cells. Conclusions: Collectively, these findings in neonatal mice suggest that LOP cooperatively enhances intestinal barrier maturation and directs stem cell differentiation via Brg1-Notch signaling, offering potential insights for future research on bioactive protein supplementation in infant nutrition. Full article