Search Results (30,082)

Duchenne muscular dystrophy (DMD) is a severe X-linked genetic disorder caused by an array of mutations in the dystrophin gene, with the most commonly mutated regions being exons 48–55. One of the several existing approaches to treat DMD is gene therapy, based on alternative splicing and mutant exon skipping. Testing of such therapy requires animal models that carry mutations homologous to those found in human patients. Here, we report the generation of two genetically modified mouse lines, named “insT” and “insG”, with distinct mutations at the same position in exon 51 that lead to a frameshift, presumably causing protein truncation. Hemizygous males of both lines exhibit classical signs of muscular dystrophy in all muscle tissues except for the cardiac tissue. However, pathological changes are more pronounced in one of the lines. Membrane localization of the protein is reduced to the point of absence in one of the lines. Moreover, an increase in full-length isoform mRNA was detected in diaphragms of insG line mice. Although further work is needed to qualify these mutations as sole origins of dissimilarity, both genetically modified mouse lines are suitable models of DMD and can be used to test gene therapy based on alternative splicing. Full article

In this study, the effect of Metarhizium spp. M. marquandii on the seed germination of cabbage, a cruciferous crop, was investigated. The effects of this strain on the seed germination vigor, bud growth and physiological characteristics of Chinese cabbage were analyzed by a seed coating method. The results showed the following: (1) The coating agent M. marquandii SGSF043 could significantly improve the germination activity of Chinese cabbage seeds. (2) The strain concentration in the seed coating agent had different degrees of regulation on the antioxidase system of the buds, indicating that it could activate the antioxidant system and improve the antioxidant ability of the buds. (3) When the concentration of M. marquandii SGSF043 was 5.6 × 10⁶ CFU/mL (average per grain), the effect of M. marquandii SGSF043 on the leaf hormones Indole Acetic Acid (IAA), Gibberellic Acid (GA) and Abscisic Acid (ABA) of Chinese cabbage seedlings was significantly higher than that of other treatment groups, indicating that the strain could optimize the level of plant hormones. (4) M. marquandii SGSF043 could induce the expression of stress-resistance-related genes in different tissue parts of Chinese cabbage and improve the growth-promoting stress resistance of buds. This study showed that M. marquandii SGSF043 could not only improve the germination vitality of Chinese cabbage seeds but also enhance the immunity of young buds. The results provide a theoretical basis for the application potential of Metarhizium marquandii in agricultural production. Full article

Members of the B-Box (BBX) family of proteins play crucial roles in the growth and development of rice. Here, we identified a rice BBX protein, Oryza sativa BBX2 (OsBBX2), which exhibits the highest expression in the root. The transcription of OsBBX2 follows a diurnal rhythm under photoperiodic conditions, peaking at dawn. Functional analysis revealed that OsBBX2 possesses transcriptional repression activity. The BBX2 was overexpressed in the rice japonica cultivar Longjing 11 (LJ11), in which Ghd7 and PRR37 were non-functional or exhibited weak functionality. The overexpression of OsBBX2 (OsBBX2 OE) resulted in a delayed heading date under a long-day (LD) condition, whereas the bbx2 mutant exhibited flowering patterns similar to the wild type (WT). Additionally, transcripts of Ehd1, Hd3a, and RFT1 were downregulated in the OsBBX2 OE lines under the LD condition. OsBBX2 interacted with Hd1 (BBX18), and the bbx2 hd1 double mutant displayed a late flowering phenotype comparable to that of hd1. Furthermore, OsBBX2 enhanced the transcriptional repression of Hd3a through its interaction with Hd1, as demonstrated in the protoplast-based assay. Taken together, these findings suggest that the OsBBX2 delays flowering by interacting with Hd1 and co-repressing Hd3a transcription. Full article

Breastfeeding is fundamental for the development and protection of the newborn, and microorganisms present in breast milk are associated with the development of the infant’s intestinal microbiota. However, there are factors that interfere with breastfeeding, resulting in the need to supply donated milk to milk banks for these children. Even though there is a restriction on medications prescribed for pregnant and breastfeeding women, some antimicrobials are accepted, as long as they are used correctly and as they can increase the selection pressure for resistant bacteria. The microorganisms present in breast milk from a human milk bank were evaluated and the resistance of the isolates to antimicrobials was phenotypically characterized. In total, 184 microbial isolates were identified by mass spectrometry, of 12 bacterial genera and 1 yeast genus. There was a high prevalence of bacteria of the genus Staphylococcus, mainly S. epidermidis (33%). Resistance to antimicrobials varied among species, with a higher percentage of isolates resistant to penicillins and macrolides. Multidrug resistance was identified in 12.6% of 143 isolates. Breast milk contains a wide variety of microorganisms, mainly those of the Staphylococcus and Enterobacter genera. There was a high percentage of resistant isolates, and multidrug resistance in Klebsiella oxytoca (66.7%; 4/6) and S. epidermidis (15.0%; 9/60) isolates, which increases the public health concern. Full article

Waste water treatment plants (WWTP) are considered as a hotspot for the acquisition and dissemination of antimicrobial resistance (AMR). The present study aimed to assess the AMR rate of the waste water microbiome in a selected WWTP and the treatment efficiency. Culture-dependent and PCR methods were used in the AMR study of raw and treated waste water (TWW) microbiomes. The population proportion of heterotrophic plate count (HPC) bacteria resistant to five antibiotic classes was assessed, as well as the AMR phenotype of a total of 238 Enterobacteriaceae and 259 Enterococcus spp. strains. Waste water treatment increased tetracycline- and ciprofloxacin-resistant bacteria by 67% and 61%, as well as the incidence of Enterobacteriaceae resistant to ciprofloxacin, co-trimoxazole, and cephalosporins. Multiple resistance increased, and 8.8% of TWW isolates exhibited an ESBL-producing phenotype, most often encoded by blaTEM and blaCTX-M genes. The most common resistance among Enterococcus spp. was to erythromycin and tetracycline, and despite the increased AMR rate among TWW isolates, only the increase in tetracycline resistance and the decrease in high-level gentamicin resistance were significant. All parameters analysed demonstrated limited removal of resistant HPC or faecal indicator bacteria in the studied WWTP and a positive selective effect towards some of them, most often to ciprofloxacin. Full article

Intra-cardiac kinetic energy (KE) and ventricular flow analysis (VFA), as derived from 4D-flow MRI, can be used to understand the physiological burden placed on the left ventricle (LV) due to bicuspid aortic valve (BAV). Our hypothesis was that the KE of each VFA component would impact the surgical referral outcome depending on LV function decrement, BAV phenotype, and aortic dilation severity. A total of 11 healthy controls and 49 BAV patients were recruited. All subjects underwent cardiac magnetic resonance imaging (MRI) examination. The LV mass was inferior in the controls than in the BAV patients (90 ± 26 g vs. 45 ± 17 g, p = 0.025), as well as the inferior ascending aorta diameter indexed (15.8 ± 2.5 mm/m² vs. 19.3 ± 3.5 mm/m², p = 0.005). The VFA KE was higher in the BAV group; significant increments were found for the maximum KE and mean KE in the VFA components (p < 0.05). A total of 14 BAV subjects underwent surgery after the scans. When comparing BAV nonsurgery vs. surgery-referred cohorts, the maximum KE and mean KE were elevated (p < 0.05). The maximum and mean KE were also associated with surgical referral (r = 0.438, p = 0.002 and r = 0.371, p = 0.009, respectively). In conclusion, the KE from VFA components significantly increased in BAV patients, including in BAV patients undergoing surgery. Full article

Lonicera caerulea L. has high nutritional and health value, and it is an important emerging small berry tree species. In this study, the morphology and nutrient composition of 60 cultivars were used to analyze and evaluate the diversity of the genus. Morphological analysis showed that the phenotypic traits of different cultivars had significant differences (p < 0.01). The phenotypic coefficient of variation (PCV) of each trait was 12.42%~84.06%, and the coefficient of genetic variation (GCV) was between 7.07%~71.72%. The analysis of nutrient content showed significant differences among the cultivars (p < 0.01). The PCV of each trait was 3.95%~96.10%, and the GCV was 0.13%~32.83%. Based on breeding objectives, cultivars with excellent growth and leaf quantitative traits, fruit quantitative traits and nutrient contents were selected through the method of comprehensive analysis of multiple characters. Traits of the selected varieties were all above average, and specific genetic gain was higher. At the same time, the selection of varieties was carried out according to flowering and fruiting phenology, which provided an indication for the breeding of improved varieties. In this study, growth, leaf and fruit quantitative traits, phenological period and nutrient components of different cultivars provided valuable information for the breeding of improved varieties. Full article

Background: The orthodontist can play an important role in the early detection of sleep-disordered breathing (SDB), aiding in the prevention of dentoskeletal complications and systemic issues. Early intervention supports proper pediatric development, emphasizing the need for SDB screening in orthodontics. SDB involves abnormal breathing during sleep, with obstructive sleep apnea (OSA) in children presenting unique diagnostic challenges compared to adults. Aim: This study aimed to identify children at risk for SDB through a validated screening questionnaire during orthodontic evaluations. Methods: This prospective study recruited children under 12 years of age between July 2023 and July 2024. The Sleep Clinical Record was used to screen for SDB indicators. Results: Among the 48 participants (31 females, 17 males) aged 5–12 years, 69% were identified as being at risk for SDB. Risk factors included oral breathing, nasal obstruction, tonsillar hypertrophy, malocclusion, high Friedman scores, narrow palates, and positive Brouillette phenotypes, all showing significant correlations (p < 0.05). Conclusion: The findings underline the critical importance of early SDB screening in orthodontic settings. These preliminary results encourage further research on larger cohorts to refine diagnostic tools and interventions. Early recognition and management of SDB can significantly enhance systemic health and craniofacial outcomes in pediatric patients. Full article

Yield-related traits have higher heritability and lower genotype-by-environment interaction, making them more suitable for genetic studies in comparison with the yield per se. Different populations have been developed and employed in QTL mapping; however, the use of reciprocal SSSLs is limited. In this study, three kinds of bi-parental populations were used to investigate the stable and novel QTLs on six yield-related traits, i.e., plant height (PH), heading date (HD), thousand-grain weight (TGW), effective tiller number (ETN), number of spikelets per panicle (NSP), and seed set percentage (SS). Two parental lines, i.e., japonica Asominori and indica IR24, their recombinant inbred lines (RILs), and reciprocal single-segment substitution lines (SSSLs), i.e., AIS and IAS, were genotyped by SSR markers and phenotyped in four environments with two replications. Broad-sense heritability of the six traits ranged from 0.67 to 0.94, indicating their suitability for QTL mapping. In the RIL population, 18 stable QTLs were identified for the six traits, 4 for PH, 6 for HD, 5 for TGW, and 1 each for ETN, NSP, and SS. Eight of them were validated by the AIS and IAS populations. The results indicated that the allele from IR24 increased PH, and the alternative allele from Asominori reduced PH at qPH3-1. AIS18, AIS19, and AIS20 were identified to be the donor parents which can be used to increase PH in japonica rice; on the other hand, IAS14 and IAS15 can be used to reduce PH in indica rice. The allele from IR24 delayed HD, and the alternative allele reduced HD at qHD3-1. AIS14 and AIS15 were identified to be the donor parents which can be used to delay HD in japonica rice; IAS13 and IAS14 can be used to reduce HD in indica rice. Reciprocal SSSLs not only are the ideal genetic materials for QTL validation, but also provide the opportunity for fine mapping and gene cloning of the validated QTLs. Full article

Arthrogryposis, which represents a group of congenital disorders, includes various forms. One such form is amyoplasia, which most commonly presents in a sporadic form in addition to distal forms, among which hereditary cases may occur. This condition is characterized by limited joint mobility and muscle weakness, leading to limb deformities and various clinical manifestations. At present, the pathogenesis of this disease is not clearly understood, and its diagnosis is often complicated due to significant phenotypic diversity, which can result in delayed detection and, consequently, limited options for symptomatic treatment. In this study, a transcriptomic analysis of the affected muscles from patients diagnosed with amyoplasia was performed, and more than 2000 differentially expressed genes (DEGs) were identified. A functional analysis revealed disrupted biological processes, such as vacuole organization, cellular and aerobic respiration, regulation of mitochondrion organization, cellular adhesion, ATP synthesis, and others. The search for key nodes (hubs) in protein–protein interaction networks allowed for the identification of genes involved in mitochondrial processes. Full article

Background: Antimicrobial resistance (AMR) is recognized as one of the most important global public health threats. There is an urgent need to reduce the spread of these multidrug-resistant bacteria (MDR-B), particularly in extremely vulnerable patients. The aim of this study was to investigate whether targeted gene amplification performed directly on clinical samples can be used simultaneously with a bundle of enhanced infection control measures in a Pediatric Intensive Care Unit (PICU) endemic to MDR-B. Methods: This study had three phases: (1) the baseline phase was performed prior to intervention when first screening and sample collection were performed; (2) the intervention phase was performed when various enhanced infection control measures (EICM) were applied; and (3) the maintenance phase occurred when EICMs were combined with the implementation of targeted molecular surveillance. The presence of four carbapenemase genes, bla_KPC, bla_OXA-48-like, bla_VIM, and bla_NDM, as well as the β-lactamase genes bla_TEM and bla_SHV, was evaluated by PCR after DNA isolation directly from stool samples. The results were compared to culture-based phenotypic analysis. Results and Conclusions: The implementation of EICM appeared to reduce the resistance burden in this sample endemic to an MDR-B clinical setting. The direct implementation of a targeted and customized rapid molecular detection assay to clinical samples seems to be an effective clinical tool for the evaluation of EICM measures. Full article

The biological process of aging is influenced by a complex interplay of genetic, environmental, and epigenetic factors. Recent advancements in the fields of epigenetics and senolytics offer promising avenues for understanding and addressing age-related diseases. Epigenetics refers to heritable changes in gene expression without altering the DNA sequence, with mechanisms like DNA methylation, histone modification, and non-coding RNA regulation playing critical roles in aging. Senolytics, a class of drugs targeting and eliminating senescent cells, address the accumulation of dysfunctional cells that contribute to tissue degradation and chronic inflammation through the senescence-associated secretory phenotype. This scoping review examines the intersection of epigenetic mechanisms and senolytic therapies in aging, focusing on their combined potential for therapeutic interventions. Senescent cells display distinct epigenetic signatures, such as DNA hypermethylation and histone modifications, which can be targeted to enhance senolytic efficacy. Epigenetic reprogramming strategies, such as induced pluripotent stem cells, may further complement senolytics by rejuvenating aged cells. Integrating epigenetic modulation with senolytic therapy offers a dual approach to improving healthspan and mitigating age-related pathologies. This narrative review underscores the need for continued research into the molecular mechanisms underlying these interactions and suggests future directions for therapeutic development, including clinical trials, biomarker discovery, and combination therapies that synergistically target aging processes. Full article

Our previous study identified the apolipoprotein M (APOM) and cytochrome P450 family 7 subfamily A polypeptide 1 (CYP7A1) genes as candidates for milk traits in dairy cattle, which were significantly up-regulated in liver tissue of Holstein cows between the dry and lactation periods. The two genes play critical roles in the peroxisome proliferator-activated receptor (PPAR) pathway. In this study, we further confirmed whether the APOM and CYP7A1 genes had significant genetic impacts on milk production traits in a Chinese Holstein population. By dual-direction sequencing of the polymerase chain reaction (PCR) products of the complete coding sequences and 2000 bp of the 5′ and 3′ flanking regions on pooled DNA sample, seven and three single nucleotide polymorphisms (SNPs) were identified in APOM and CYP7A1, respectively. With SAS 9.2, phenotype-genotype association analysis revealed such SNPs were significantly associated with at least one of the milk production traits, including 305-day milk yield, milk fat yield, milk fat percentage, milk protein yield, and milk protein percentage in the first and second lactations (p = <0.01~0.04). With Haploview 4.2, we further found that six SNPs in APOM and thee SNPs in CYP7A1 formed one haplotype, respectively. The haplotypes were significantly associated with at least one of milk production traits as well (p = <0.01~0.02). Of note, we found the SNPs in the 5′ regulatory region, rs209293266 and rs110721287 in APOM and rs42765359 in CYP7A1, significantly impacted the gene transcriptional activity after mutation (p < 0.01) through changing the transcription factor binding sites by using luciferase assay experiments. Additionally, with RNAfold Web Server, rs110098953 and rs378530166 changed the mRNA secondary structures of APOM and CYP7A1 genes, respectively. In summary, our research is the first to demonstrate that APOM and CYP7A1 genes have significantly genetic effects on milk yield and composition traits, and the identified SNPs may serve as available genetic markers for genomic selection program in dairy cattle. Full article

The societal pressure on intensive pastoral dairying demands the search for strategies to reduce the amount of N flowing through and excreted by dairy cows. One of the strategies that is being currently explored focuses on the animal as a solution, as there are differences in N metabolism between cows even within the same herd. This work was conducted to explore such an approach in A1PF herds in New Zealand and the possibility of identifying A1PF cows that are divergent for milk urea nitrogen (MUN) concentration through phenotyping as a potential viable strategy to reduce N leaching and emissions from temperate dairy systems. Three herd tests were conducted to select a population sample of 200 cows (exhibiting the lowest 100 and highest 100 MUN concentrations). Milk samples were collected from the 200 cows during mid and late lactation to test for milk solids content and MUN. From the 200 cows, urine for urinary N concentration (UN), blood for plasma urea N, total antioxidants (TAS), and glutathione peroxidase (GPx) were collected from the 20 extremes (the lowest 10 and highest 10 MUN concentrations). Milk urea N was greater in cows selected as high-MUN cows (16.2 vs. 14.32 ± 0.23 mg/dL) and greater during late lactation (16.9 vs. 13.0 ± 0.19 mg/dL). Milk solids and fat content were 38% and 20% greater in cows selected as low-MUN cows than in high-MUN cows during mid lactation (p < 0.001). Low-MUN cows had lower UN than high-MUN cows during mid lactation (0.64 vs. 0.88 ± 0.11%). The N concentration in the plasma (p = 0.01) and Tas (p = 0.06) were greater during late lactation. There was a positive relationship between the MUN concentration phenotype used for selection and the MUN concentration for the trial period and MUN concentration and UN concentration during mid and late lactation (p < 0.001). Our results suggest that A1PF cows within a commercial herd can be phenotyped and selected for low-MUN, which may be potentially a viable strategy to reduce N losses to the environment and create healthier systems. Following genetic tracking, those cows can be bred to further promote low-MUN A1PF herds. Full article

Macrophages undergo polarization, resulting in distinct phenotypes. These transitions, including de-/repolarization, lead to hysteresis, where cells retain genetic and epigenetic signatures of previous states, influencing macrophage function. We previously identified a set of interferon-stimulated genes (ISGs) associated with high lipid levels in macrophages that exhibited hysteresis following M1 polarization, suggesting potential alterations in lipid metabolism. In this study, we applied weighted gene co-expression network analysis (WGCNA) and conducted comparative analyses on 162 RNA-seq samples from de-/repolarized and lipid-loaded macrophages, followed by functional exploration. Our results demonstrate that during M1 hysteresis, the sustained high expression of Marco (SR-A6) enhances lipid uptake, while the suppression of Abca1/2 reduces lipid efflux, collectively leading to elevated intracellular lipid levels. This accumulation may compensate for reduced cholesterol biosynthesis and provide energy for sustained inflammatory responses and interferon signaling. Our findings elucidate the relationship between M1 hysteresis and lipid metabolism, contributing to understanding the underlying mechanisms of macrophage hysteresis. Full article