Search Results (2,897)

Glaesserella parasuis (G. parasuis) is a key pathogen responsible for swine respiratory disease, and the development of broadly protective vaccines is hampered by its high antigenic diversity. The iron-acquisition protein TbpB is a conserved vaccine candidate, but the cellular immune responses it elicits, particularly T-cell subset dynamics during immunization and challenge, remain insufficiently defined. This study characterized these responses after oral immunization of colostrum-deprived piglets with the TbpB^Y167A mutant. Ten colostrum-deprived piglets were allocated to immunized and non-immunized (PBS) groups, immunized at days 15 and 30 of life and subsequently challenged with G. parasuis (45 days old); peripheral blood mononuclear cells were collected at baseline, after each immunization, and at 1 and 3 days post-infection. Multiparametric flow cytometry was used to quantify major leukocyte subsets and T-cell phenotypes defined by sIgM, CD172a, CD3, TCRγδ, CD8α/β, CD4 and CD27 expression. Booster immunization induced significant expansion of B cells (p < 0.01), TCRγδ T cells (p < 0.01), CD8⁺ αβ T cells (p < 0.001) and CD4⁺ memory T cells (p < 0.01) in immunized piglets compared with controls. After challenge, CD8⁺ cytotoxic T cells in immunized animals rapidly shifted from naïve to memory phenotypes, peaking at 48–72 h (p < 0.01). These biphasic T-cell dynamics are consistent with the protective efficacy previously demonstrated for this vaccine in colostrum-deprived piglets, and support a key contribution of TCRγδ, CD8⁺ cytotoxic and CD4⁺ memory T cells to immunity against G. parasuis and to the design of next-generation vaccines. Full article

Background: The UGT1A1 gene is associated with the toxicity caused by SN38, the cytotoxic component of Sacituzumab govitecan (SG) used in the treatment of metastatic triple-negative breast cancer (mTNBC), among other approved indications. In this study, we aimed to analyze the effect of UGT1A1*28 allele on the safety and, secondarily, the effectiveness of SG in mTNBC. Methods: This was a multicenter, ambispective study that included patients treated with SG for mTNBC. Genotyping for UGT1A1*28 was performed using real-time polymerase chain reaction (PCR). Adverse events (AEs) of grade ≥ 2 during the first three cycles were compared between patients who were homozygous mutant (UGT1A1*28/*28) and those with wild-type (WT) or heterozygous genotypes. Effectiveness between the two groups was also compared using progression-free survival (PFS) and overall survival (OS) assessed with the Kaplan–Meier method. Results: A total of 81 patients were included: 37.0% were WT, 55.6% heterozygous, and 7.4% homozygous mutant. All UGT1A1 *28/*28 patients experienced grade ≥ 2 AEs (100% vs. 69.3%; p = 0.109), with a statistically significant association in the case of febrile neutropenia (33.3% vs. 6.7%; p = 0.025), and a trend towards higher rates of anemia and diarrhea (50.0% vs. 17.3%; p = 0.053). Genotype did not influence PFS or OS; however, dose reductions were associated with better survival outcomes. Conclusions: This real-world study shows a correlation between toxicity and the presence of the UGT1A1*28 mutation in patients treated with SG for mTNBC. Improving treatment tolerability through dose reductions may enhance SG effectiveness. These findings support the implementation of UGT1A1 genotyping in routine clinical practice. Full article

Background and Objectives: Our study aimed to investigate the effect of RAS and TP53 mutations, either alone or in combination, on survival in patients with metastatic colorectal carcinoma (mCRC). Materials and Methods: Patients diagnosed with mCRC and followed up at our center between January 2019 and November 2023, who underwent somatic mutation analysis via next-generation sequencing (NGS) testing, were retrospectively evaluated. A total of 155 patients were evaluated within the scope of the study. Patients were grouped as mutant type (m) or wild type (w) for RAS and TP53. Survival times between the groups were examined using the Kaplan–Meier method. Cox regression analysis was performed for factors with a prognostic effect on survival. Results: Among the patients, 35.4% exhibited an RASm/TP53w mutation profile, 30.9% had RASw/TP53w, 20% had RASw/TP53m, and 13.5% had RASm/TP53m. The lowest median progression-free survival (mPFS) and median overall survival (mOS) durations were observed in the RASm/TP53w group (7.3 months and 16.9 months, respectively). Median OS was significantly lower in the RASm/TP53w group compared to the RASw/TP53w group (16.9 months vs. 26.0 months, p = 0.003), whereas no significant difference was found between mPFS durations. No statistically significant difference was observed between the RASw/TP53m and RASm/TP53m groups and the RASw/TP53w group for mPFS and mOS. The RASm/TP53w mutation profile was identified as an independent prognostic factor for decreased OS in the multivariate Cox regression analysis. Conclusions: In mCRC cases with the RASm/TP53w mutation profile, the mOS was significantly lower. The RASm/TP53w mutation profile was identified as an independent prognostic factor for decreased OS. These findings are expected to contribute to the literature as real-world evidence regarding the prognostic value of different RAS and TP53 mutation combinations in mCRC. Full article

Vascular endothelial growth factor receptor 3 (VEGFR3) assumes a pivotal role in regulating the development and maintaining the structural integrity of the lymphatic system. Decreased activity of VEGFR3 can precipitate aplasia or hypoplasia of lymphatic system components, culminating in primary lymphedema. To date, numerous genetic variants have been identified within the FLT4 gene, which encodes VEGFR3; however, the majority of these remain uncharacterised and are classified as ‘variants of uncertain significance’. In preceding investigations involving FLT4 sequence analysis conducted on individuals presenting with primary lymphedema, we identified several rare genetic variants that possess the potential to modulate the functional activity of VEGFR3, including the heterozygous variant c.3175G>C (p.A1059P). Preliminary assessments encompassing clinical characteristics, family history, and predictive computational algorithms indicated that this variant was likely pathogenic. Consequently, this study presents the results of functional evaluation of the mutant VEGFR3 activity in cell models overexpressing the FLT4 variant c.3175G>C. VEGFC-dependent VEGFR3 phosphorylation and FLT4 expression were reduced in cells with c.3175G>C FLT4 variant compared to wild-type, confirming the pathogenic role of c.3175G>C in primary lymphedema. Full article

Salmonella enterica from low-moisture food has been found to have a higher thermal tolerance than from high-moisture food. However, the molecular mechanism underlying the association of thermal tolerance of this pathogen with low-moisture foods, such as peanut butter and peanut spread, has not been fully elucidated. We previously found that mutants of S. Tennessee with a defective gene encoding a cell membrane lipoprotein (Lpa) or cell division protein (ZapC) formed significantly (p ≤ 0.05) less biofilm than the wildtype strain. To assess the possible role of these genes in the thermal tolerance of S. Tennessee, this study compared the surviving populations of the wildtype S. Tennessee and its mutants defective in Lpa or ZapC in different types of peanut products (regular, reduced-fat, and natural) at 74 °C for 0, 2.5, 5, 10, 15, 20, 30, 40, or 50 min. Results showed that mutants with a defective lpa or zapC significantly affected the survival of Salmonella in peanut products during heat treatments. Significantly, a higher reduction in Salmonella population was observed in regular peanut butter, followed by natural and reduced-fat peanut spreads. The study provides new insight into one of the molecular mechanisms underlying the thermal tolerance of Salmonella enterica. Full article

Breast cancer and lung adenocarcinoma share common features, including female predominance and the expression of estrogen receptor (ER) and epidermal growth factor receptor (EGFR) during carcinogenesis. Patients with breast cancer have a significantly higher risk of developing second primary lung cancer than those without breast cancer. ER beta expression is associated with resistance to EGFR tyrosine kinase inhibitors (TKIs) in EGFR-mutant lung adenocarcinoma, indicating a potentially important interaction between ER and EGFR. However, the mechanisms underlying this crosstalk remain poorly understood. Our clinical data showed a significant correlation between antiestrogen treatment for breast cancer and mutant EGFR expression (p = 0.021) in lung adenocarcinoma patients. In vitro, tamoxifen upregulated phosphorylated EGFR (p-EGFR) in EGFR-mutant lung adenocarcinoma cell lines. Heparin-binding EGF-like growth factor was identified as a key mediator from the ER pathway that stimulates p-EGFR. Tamoxifen counteracts estrogen’s effect and restores p-EGFR upregulation. Furthermore, coadministration of tamoxifen and the EGFR TKI gefitinib potentially inhibited p-EGFR expression in EGFR-mutant lung adenocarcinoma. Regular follow-up with chest computed tomography is recommended for patients with breast cancer. For those diagnosed with both ER-positive breast cancer and EGFR-mutant lung adenocarcinoma, combined tamoxifen and EGFR TKI therapy may offer an effective targeted treatment strategy. Full article

Background/Objectives: FLOURY ENDOSPERM 2 (FLO2) is known to affect rice endosperm development and starch quality, yet its role in determining flour physicochemical behavior and endosperm structural integrity has not been quantitatively defined. This study aimed to elucidate how loss of FLO2 function alters starch organization and functional properties of rice flour. Methods: Two independent homozygous, T-DNA-free OsFLO2 knockout lines were generated in the japonica cultivar Dongjin using CRISPR/Cas9. Grain appearance was evaluated in mature seeds. Flour physicochemical properties were analyzed by Rapid Visco Analyzer (RVA) and differential scanning calorimetry (DSC). Amylopectin chain-length distribution was determined by isoamylase debranching followed by HPAEC-PAD, and endosperm microstructure was examined by scanning electron microscopy. Results: OsFLO2 mutants exhibited floury, opaque endosperms, with chalkiness increasing from 4.8% in the WT to 27–29%. RVA analysis showed a marked reduction in peak viscosity (1193 cP to 263–293 cP) and a decrease in pasting temperature (77.2 °C to 68.9–70.5 °C). DSC indicated a tendency toward reduced gelatinization enthalpy in the mutants. These changes were associated with a reduced proportion of short amylopectin chains (DP 6–12), decreased long chains (DP ≥ 37), and a relative increase in intermediate-long chains (DP 25–36), along with disrupted granule packing and a 1.33–1.36-fold increase in endosperm porosity. Conclusions: These results demonstrate that FLO2 plays an important role in maintaining the structural integrity of rice endosperm by harmonizing the microstructure of amylopectin with the thermal and gelatinization properties of starch. Full article

The Na⁺/K⁺/2Cl⁻ cotransporter (NKCC) gene encodes a critical membrane transporter involved in cellular ion homeostasis and plays a pivotal role in osmoregulation and salinity adaptation in aquatic organisms. This study identified and validated SNP markers in the NKCC gene associated with low-salinity tolerance in Scylla paramamosain. Four SNPs (g.196C>A, g.8374T>A, g.8385T>A and g.91143T>A) were screened and genotyped in low-salinity tolerant and intolerant groups. Association analysis revealed that mutant genotypes at all four sites were significantly enriched in the tolerant group (p <0.05), with the values of odds ratios (OR) greater than 1. The tolerant group exhibited significantly higher genetic diversity than the intolerant group. Haplotype analysis showed the wild CTTT haplotype dominated in the intolerant group, while mutant-containing haplotypes were significantly elevated in the tolerant group. A positive correlation was observed between the mutant and NKCC expression. Functional validation by qRT-PCR demonstrated that mutant allele carriers exhibited significantly higher NKCC mRNA expression levels than the wild-type carriers. Moreover, the expression level of homozygous mutations is significantly higher than that of heterozygous mutations. These validated SNPs could provide effective molecular markers for marker-assisted selection breeding of low-salinity tolerant S. paramamosain strains, offering important theoretical and practical implications for sustainable aquaculture development. Full article

Background: IDH1-mutant oligodendroglioma and astrocytoma differ not only in growth rate but also in growth pattern. Oligodendrogliomas tend to infiltrate white matter tracts, whereas astrocytomas more often displace them. Such difference could lead to different cognitive outcomes. This study examined differences in executive functioning before and up to one year after surgery between patients with IDH1-mutant astrocytoma and oligodendroglioma. Methods: Patients with WHO grade 2–3 IDH1-mutant oligodendroglioma (1p19q-codeleted) or astrocytoma were included. Cognition was assessed preoperatively, and at 3 and 12 months postoperatively using standardized computerized and paper-and-pencil tests. Groups were compared on demographics, tumor characteristics, surgical modality, extent of resection, adjuvant treatment, and baseline cognition. Longitudinal mixed models were performed to investigate differences in performances over time for the total sample and stratified by surgical approach (awake vs. asleep). Results: 162 patients (67 oligodendroglioma, 95 astrocytoma) were included. Oligodendroglioma patients were older, with more frontal and fewer temporal tumors. Oligodendroglioma patients showed a greater impairment prevalence on a measure of inhibition before surgery. In the awake surgery group, no longitudinal differences were found between diagnoses. In the asleep surgery group, astrocytoma patients remained stable while oligodendroglioma patients declined on a measure of cognitive flexibility, with performance at 3 and 12 months significantly lower than at baseline. Conclusions: Specific aspects of executive functioning in IDH1-mutant gliomas may differ by subtype. Oligodendroglioma patients showed postoperative decline in cognitive flexibility that did not recover to baseline level, particularly in case of surgery under general anesthesia. These results highlight the potential relevance of tumor subtype and surgical approach in limiting cognitive risks after glioma surgery. Full article

Thrombospondin-1 (TSP1) is a multifunctional glycoprotein that plays a crucial role in angiogenesis and vascular remodeling. Ser93 of TSP1 has recently been identified as a novel phosphorylation site, influencing angiogenic properties; however, the underlying signaling mechanism remains unclear. Here, we investigated the functional impact of Ser93 phosphorylation using phosphomimetic (TSP1^S93D) and phosphonull (TSP1^S93A) mutants. Endothelial cell (EC) migration was analyzed using scratch assay and electric cell-substrate impedance sensing. Activation of key pathways (Akt, p38, ERK, and FAK) was analyzed by immunoblotting. TSP1 secretion was quantified by ELISA. Downstream effects on smooth muscle cells were examined by Western blot using conditioned media of endothelial cells. Expression of TSP1^S93D significantly impaired endothelial migration and wound closure, associated with reduced phosphorylation of FAK and paxillin. Upstream of FAK signaling, TSP1^S93D showed enhanced binding to integrin β1 and promoted its clustering. In contrast, TSP1^S93D stimulated smooth muscle cell proliferation, migration, cytoskeletal remodeling, and phenotypic switching toward a synthetic, pro-inflammatory state characterized by elevated marker protein expression. Together, these findings demonstrate that the impaired angiogenic properties induced by TSP1^S93D result from the modulation of integrin β1-FAK pathways in ECs, suppressing endothelial motility while promoting smooth muscle activation, suggesting a role in early vascular remodeling and inflammation. Full article

Group A rotaviruses (RVAs) remain the leading cause of acute gastroenteritis (AGE) in young children in low- and middle-income countries. In Brazil, the oral attenuated RVA vaccine (Rotarix^®), monovalent genotype G1P[8], is distributed by the national immunization program and has drastically reduced morbidity and mortality associated with RVA etiology. In this study, Rotarix^® G1P[8] was detected using specific qRT-PCR from the fecal shedding of children living in the Amazon region, and 18.3% (29/158) were positive and 75.8% (22/29) presented with AGE. The VP4 (VP8*) gene of these sheddings, submitted to Sanger nucleotide sequencing, showed an occurrence of mutations, including the silent mutation at 144C > G (one child) and the following missense mutations— 499T > C (F167L) (two children), 644G > C (C215S) (one child), and 787G > A (E263K) (one child). These mutations had no impact on the protein model structure in silico deduced from the VP4 (VP8*) mutants. The in silico protein model deduced from the VP4 (VP8*) nucleotide sequences, bound to type 1H sugar antigens (H1) and its precursor Lac-para-N-biose (LNB), had a stronger binding to the G1P[8] genotype, when compared to G3P[8]. Rotarix^® shedding was higher in HBGA secretors than in non-secretors (79.3%; 23/29). A total of 11.4% (18/158) of children with Rotarix^® G1P[8] shedding were unvaccinated, indicating the occurrence of indirect protection. Stability evidence of Rotarix^® VP4 (VP8*) spike protein from samples collected in vivo is presented. Full article

Systems favoring cross-pollination, such as male sterility and female flowering type, are of great importance in the development of new hybrid cultivars and their seed production. The advantages of male sterility are expressed in the production of cheaper and competitive seeds. The presence of this characteristic in watermelon is not common, and in some cases, it is accompanied by negative manifestations. A collection of 150 watermelon genotypes was tested at the Maritsa Vegetable Crops Research Institute, Bulgaria, over the past nine years to search for a genetic source of male sterility. The results revealed that two mutations were found. The first mutation was in a plant of the Asar variety, which formed completely degenerated structures in the place of male and female flowers that were completely sterile. The other mutation affected male flowers, female flowers, and leaf shape. Male flowers produced a small amount of pollen. Female flowers were formed, but they were sterile and aborted at an early stage. The genotype can be propagated by pollination of the normal plants, which in the next generation segregate into mutant—25% and normal—75%. The gene source is phenotyped according to the main characteristics of the fruits and the vegetation period. The mutation found cannot be directly used in a breeding program, but it is of interest for studying this important trait. The success of detecting flowers that are sterile depends on the number of watermelon plants, which, for the conditions of the experiment, amounted to a minimum of 4492 plants at a probability level of P₃—0.95. Full article

Astaxanthin, one of the most commercially valuable carotenoids, is renowned for its potent antioxidant and anti-inflammatory properties and is experiencing growing demand across diverse industries. To enhance astaxanthin production in Paracoccus marcusii, compound mutagenesis was performed using ethyl methanesulfonate (EMS), ultraviolet (UV) radiation, and atmospheric room temperature plasma (ARTP) treatment. Subsequently, a high-throughput microbial microdroplet culture (MMC) system was employed to select fast-growing microdroplet, followed by screening for high astaxanthin-producing mutants on dual-inhibitor plates. The mutant M21 was isolated and exhibited a significant increase of 16.86% in astaxanthin content (1.53 mg/g) and a 19.81% increase in astaxanthin production (11.71 mg/L) compared with the wild type (WT) (p < 0.05). Moreover, the enhanced phenotype of M21 was genetically stable. Response surface methodology (RSM)-based optimization of fermentation conditions further increased astaxanthin content and production to 1.72 mg/g and 12.92 mg/L, respectively, corresponding to improvements of 16.44% and 23.02% over the WT, while simultaneously reducing culture time, total nitrogen requirements, and sodium lactate consumption, thereby lowering production costs. This study achieved significant enhancement of astaxanthin production through novel mutant breeding and fermentation optimization, underscoring the effectiveness of this integrated strategy for application in industrial biotechnology. Full article

Altered protein synthesis plays a key role in ageing and multiple neurodegenerative diseases. In Alzheimer’s disease and other tauopathies, the intracellular accumulation of hyperphosphorylated Tau disrupts several cellular processes, including mRNA translation. Although Tau interacts with ribosomal proteins and modulates translational selectivity, its effects on global protein synthesis remain poorly understood. Studies report reduced translation in later disease stages but increased translation early in pathology. To clarify Tau’s impact in human neurons, we used SH-SY5Y cells overexpressing the P301L mutant form of Tau and quantified global protein synthesis using the SUnSET (Surface Sensing of Translation) puromycin-incorporation assay. We found that Tau-P301L expression greatly increased global translation by upregulating mTOR/S6 pathway. These effects were abolished by rapamycin treatment, indicating that Tau-driven translational upregulation is mTOR-dependent. Given that impaired translational control can disrupt synaptic plasticity and memory, Tau-induced alterations in protein synthesis may contribute to tauopathy progression and identify mTOR signalling as a potential therapeutic target. Full article

Chemorepulsion mechanisms for eukaryotic cells are poorly understood. We performed proteomics and phosphoproteomics to elucidate how Dictyostelium discoideum responds to its two endogenous chemorepellent signals, the protein AprA and inorganic polyphosphate (polyP). AprA and polyP affected levels of more than 200 proteins, with an overlap of both upregulating 25 proteins and downregulating two proteins. Two proteins were upregulated by AprA but downregulated by polyP, while two others showed the opposite trend. Surprisingly, many of the AprA- and polyP-regulated proteins are associated with RNA metabolism and ribosomes. AprA increased phosphorylation of 15 proteins and decreased phosphorylation of 36 proteins. PolyP increased phosphorylation of 12 proteins and decreased phosphorylation of 12 proteins. As expected, the two chemorepellents affected phosphorylation of signal transduction/ motility proteins, but unexpectedly affected phosphorylation of RNA-associated proteins. Both AprA and polyP decreased phosphorylation of five proteins including the Ras-interacting protein RipA and guanine nucleotide exchange factors (GEFs) such as the RacGEF GxcT. Mutants lacking RipA or GxcT were unresponsive to both AprA and polyP chemorepulsion. Together, this work supports the idea that rather than activating the same chemorepulsion mechanism, AprA and polyP activate only partially overlapping chemorepulsion mechanisms, and identifies two new components that are used by both chemorepellents. Full article