Search Results (38)

Background/Objectives: The X-inactivation specific transcript (XIST) is a long noncoding RNA playing a crucial regulatory role in X chromosome inactivation (XCI)—a transcriptional regulatory process that silences one of the two X chromosomes in females to ensure proper dosage compensation between male and female mammals. The transcription of XIST is maintained throughout a female’s lifespan in all somatic cells, where XIST RNA binds to the X chromosome in cis and ensures chromosome-wide gene silencing. Disrupting XIST expression can lead to transcriptional reactivation of X-linked genes and epigenetic changes affecting cell development. The prevalence of XIST regulatory effects on mammalian transcription, however, remains unclarified. Methods: Here we performed a comparative expression analysis using RNA-sequencing datasets from recently published studies and examined the consequences of XIST-deletion on transcription at the whole genome, individual chromosome, and specific gene levels. We investigated the common differentially expressed genes (DEGs) and biological pathways following XIST loss across cell types, together with differential transcriptional analysis comparing the X chromosome and autosomes using cumulative distribution fractions. We analyzed the distribution of DEGs along the X chromosome with scatterplots and correlation analysis incorporating gene density and transposable elements. Results: Our findings indicate that the loss of XIST causes transcriptional changes in the X chromosome and autosomes that differ depending on cell type and state. XIST-deletion results in differential expression of genes subject to XCI-silencing as well as genes escaping XCI. In all the cell types we analyzed, X-linked genes show differential expression across the entire X chromosome in a cluster-like pattern according to gene density and, in certain cell types, correlate strongly with short interspersed nuclear element (SINE) distributions. Conclusions: Our results demonstrate that transcriptional roles of XIST can be highly associated with cell state: stem cells have different transcriptional responses compared to differentiated cells following XIST loss. Full article

Background: Bacterial Chondronecrosis with Osteomyelitis (BCO) is a significant issue affecting the welfare and economy of the broiler industry, causing substantial revenue losses annually. This disease is frequently associated with Staphylococcus spp. and Enterococcus spp. infections and necrosis of leg and vertebral bones. The typical annual lameness incidence of approximately 3–5% may increase to 30% during outbreaks. Neither the etiology or pathogenesis of the disease has been comprehended, nor have effective preventative measures been identified. Electron beam (eBeam) technology is renowned for producing efficient whole-cell vaccines by preventing bacterial multiplication through irreversible DNA shredding while preserving the integrity of membrane proteins (immunogenic epitopes). This study aims to reduce BCO-induced lameness in broiler chickens via in ovo immunization using eBeam-inactivated multi-strain Staphylococcus. Methods: A total 1080 birds were assigned to four vaccination groups: eBeam-inactivated, formalin-inactivated, combination of eBeam- and formalin-inactivated, and sham (vehicle). The birds were directly exposed to aerosolized, natural BCO challenge until 56 days of age. Results: Birds vaccinated with the eBeam-inactivated Staphylococcus vaccine showed a significant reduction (>50%) in daily cumulative lameness compared to other groups and a decrease in Staphylococcus colonization was observed in the leg joints of treated birds. Conclusions: the eBeam-inactivated Staphylococcus vaccine successfully prevented BCO lameness in broiler chickens. Full article

Background: Vaccination is often a highly effective approach for protecting against clinical disease and mortality caused by viruses. However, vaccine efficacy against viral transmission has rarely been assessed, which can provide vital information on the eradication efficacy and sustainability of vaccines in the field. Methods: Here, we evaluated the host mortality, shedding, and direct fish-to-fish transmission protection efficacy of three vaccine regimens (DNA, inactivated, and attenuated) against infectious hematopoietic necrosis virus (IHNV) in rainbow trout. We quantified protection against single- and mixed-genotype IHNV infections when the vaccines were delivered by intramuscular injection, intraperitoneal injection, and bath immersion, respectively, to reflect field conditions. Results: All three vaccine regimens provided significant protection against fish mortality. The DNA vaccine regimen was qualitatively the most protective and the attenuated vaccine regimen the least. However, these three vaccines provided limited protection against viral shedding. Cumulative shedding over the course of the infection was only slightly reduced compared to unvaccinated fish. There was some indication that the viral genotype fish were exposed to influenced vaccine efficacy, perhaps as a result of genetic similarity to the vaccine strain. Likewise, the DNA vaccine reduced direct transmission in fish cohabitation experiments from 100% to 50%. The inactivated and attenuated vaccine had little impact on IHNV transmission. Conclusions: Collectively, our results suggest that existing IHNV vaccines that increase host survival provide minimal virus transmission protection in rainbow trout, which is likely to limit their long-term efficacy in the field. This work contributes to a growing body of evidence that enhancement of the transmission protection of IHNV and other vaccines will likely bolster disease reduction in the field. Full article

Cumulative inhibition of voltage-gated Na⁺ channel current (I_Na) caused by high-frequency depolarization plays a critical role in regulating electrical activity in excitable cells. As discussed in this review paper, exposure to certain small-molecule modulators can perturb I_Na during high-frequency stimulation, influencing the extent of cumulative inhibition and electrical excitability in excitable cells. Carbamazepine differentially suppressed transient or peak (I_Na(T)) and late (I_Na(L)) components of I_Na. Moreover, the cumulative inhibition of I_Na(T) during pulse-train stimulation at 40 Hz was enhanced by lacosamide. GV-58 was noted to exert stimulatory effect on I_Na(T) and I_Na(L). This stimulated I_Na was not countered by ω-conotoxin MVIID but was effectively reversed by ranolazine. GV-58′s exposure can slow down I_Na inactivation elicited during pulse-train stimulation. Lacosamide directly inhibited I_Na magnitude as well as promoted this cumulative inhibition of I_Na during pulse-train stimuli. Mirogabalin depressed I_Na magnitude as well as modulated frequency dependence of the current. Phenobarbital can directly modulate both the magnitude and frequency dependence of ionic currents, including I_Na. Previous investigations have shown that exposure to small-molecule modulators can perturb I_Na under conditions of high-frequency stimulation. This ionic mechanism plays a crucial role in modulating membrane excitability, hereby supporting the validity of these findings. Full article

Sutures provide mechanical support for wound closure after various traumas and surgical operations. Absorbable sutures are increasingly favored as they eliminate the need for secondary procedures and minimize additional damage to the wound site. In this study, chitosan sutures were produced using the dry jet–wet spinning method, achieving number 7-0 sutures (approximately 76 μm diameter) with a homogeneous surface. FTIR analysis demonstrated molecular interactions between chitosan and TiO₂ or curcumin, confirming successful incorporation. The addition of 3% TiO₂ increased the tensile strength of chitosan sutures by 12.32%, reaching 189.41 MPa. Morphological analysis revealed smooth surfaces free of pores and bubbles, confirming the production of high-quality sutures. Radical scavenging activity analysis showed that curcumin-loaded sutures exhibited 43% scavenging ability after 125 h, which was significantly higher compared to pure chitosan sutures. In vitro antibacterial tests demonstrated that curcumin-loaded sutures provided 98.87% bacterial inactivation against S. aureus within 24 h. Additionally, curcumin release analysis showed a cumulative release of 77% over 25 h. The bioactivity of the sutures was verified by hydroxyapatite layer formation after incubation in simulated body fluid, supporting their potential for tissue regeneration. These findings demonstrate that TiO₂ reinforcement and curcumin loading significantly enhance the functional properties of chitosan sutures, making them strong candidates for biocompatible and absorbable surgical applications. Full article

Background: Monoamine oxidases (MAO) are flavoenzymes that metabolize a range of brain neurotransmitters, whose dysregulation is closely associated with the development of various neurological disorders. This is why MAOs have been the central target in pharmacological interventions for neurodegeneration for more than 60 years. Still, existing drugs only address symptoms and not the cause of the disease, which underlines the need to develop more efficient inhibitors without adverse effects. Methods: Our drug design strategy relied on docking 25 organic scaffolds to MAO-B, which were extracted from the ChEMBL20 database with the highest cumulative counts of unique member compounds and bioactivity assays. The most promising candidates were substituted with the inactivating propargylamine group, while further affinity adjustment was made by its N-methylation. A total of 46 propargylamines were submitted to the docking and molecular dynamics simulations, while the best binders underwent mechanistic DFT analysis that confirmed the hydride abstraction mechanism of the covalent inhibition reaction. Results: We identified indole-2-propargylamine 4fH and indole-2-N-methylpropargylamine 4fMe as superior MAO-B binders over the clinical drugs rasagiline and selegiline. DFT calculations highlighted 4fMe as more potent over selegiline, evident in a reduced kinetic requirement (ΔΔG^‡ = −2.5 kcal mol⁻¹) and an improved reaction exergonicity (ΔΔG_R = −4.3 kcal mol⁻¹), together with its higher binding affinity, consistently determined by docking (ΔΔG_BIND = −0.1 kcal mol⁻¹) and MM-PBSA analysis (ΔΔG_BIND = −1.5 kcal mol⁻¹). Conclusions: Our findings strongly advocate 4fMe as an excellent drug candidate, whose synthesis and biological evaluation are highly recommended. Also, our results reveal the structural determinants that influenced the affinity and inhibition rates that should cooperate when designing further MAO inhibitors, which are of utmost significance and urgency with the increasing prevalence of brain diseases. Full article

This survey reviews modern ideas on the structure and functions of mitochondrial and cytosolic aconitase isoenzymes in eukaryotes. Cumulative experimental evidence about mitochondrial aconitases (Aco2) as one of the main targets of reactive oxygen and nitrogen species is generalized. The important role of Aco2 in maintenance of homeostasis of the intracellular iron pool and maintenance of the mitochondrial DNA is discussed. The role of Aco2 in the pathogenesis of some neurodegenerative diseases is highlighted. Inactivation or dysfunction of Aco2 as well as mutations found in the ACO2 gene appear to be significant factors in the development and promotion of various types of neurodegenerative diseases. A restoration of efficient mitochondrial functioning as a source of energy for the cell by targeting Aco2 seems to be one of the promising therapeutic directions to minimize progressive neurodegenerative disorders. Full article

Motile aeromonad septicemia (MAS), caused by the Aeromonas species, has been a serious problem in fish health management, particularly in Nile tilapia (Oreochromis niloticus). This study characterized an Aeromonas species isolated from farmed tilapia fingerlings in Binangonan, Rizal, Philippines, and tested for its pathogenicity in tank trials. The isolate, designated as Aeromonas veronii DFR01 (Diseased Fish Rizal), was identified based on 16S rRNA phylogenetic analysis, 16S rRNA homology, and MALDI-TOF mass spectrometry. Its biochemical profile was generated from API and Biolog Gen III systems. A median lethal dose of A. veronii DFR01 was determined to be 10⁷ CFU/mL in tank trials and was utilized as a whole-cell inactivated antigen for oral vaccine development. The immunized tilapia fingerlings produced elevated levels of immunoglobulin M (IgM) in the blood as determined by an enzyme-linked immunosorbent assay (ELISA). There was a significant increase in IgM levels 14 days post-vaccination. A quantitative polymerase chain reaction (qPCR) showed increasing levels of IgM gene expression after vaccination until 38 days of culture. Vaccinated fish showed 25–35% cumulative mortality after the challenge, while non-vaccinated-challenged fish showed 75% mortality. The findings of this research suggest that the fish oral vaccine may prove beneficial for farmed tilapia populations. The vaccine elicited improved immune responses in the fish and resulted in higher survival rates. Full article

We previously reported the first-in-human assessment of three doses (2, 5, and 10 µg) of purified inactivated Zika virus vaccine (PIZV or TAK-426) in the Phase 1 ZIK-101 study (NCT03343626). Here, we report dose selection based on extended safety and immunogenicity data (6 months post-vaccination) and discuss considerations (e.g., immunological, historic, flavivirus immunological cross-reactions) for selecting a Zika virus (ZIKV) vaccine dose formulation. TAK-426 dose selection was conducted at the first interim analysis, and was based on cumulative safety data from both flavivirus-naïve (up to ≥28 days post-dose PD2) and flavivirus-primed participants (up to ≥28 days PD1), and on immunogenicity data from flavivirus-naïve participants only (at 28 days PD1 and 28 days PD2). The safety profile from TAK-426 recipients was compared to placebo recipients. Immunogenicity was assessed by geometric mean titer ratios of neutralizing anti-ZIKV antibodies and differences in seroconversion rates. There was no significant difference in safety between the three TAK-426 doses. The 10 μg dose provided the earliest and strongest immune response (with close to 100% seroconversion and higher antibody titers PD1 in flavivirus-naïve participants), and was well tolerated with acceptable safety profiles in both flavivirus-naïve and flavivirus-primed participants; this dose was selected for further development. Full article

Background and Objectives: Aberrant upregulation of fatty acid synthase (FASN), catalyzing de novo synthesis of fatty acids, occurs in various tumor types, including human hepatocellular carcinoma (HCC). Although FASN oncogenic activity seems to reside in its pro-lipogenic function, cumulating evidence suggests that FASN’s tumor-supporting role might also be metabolic-independent. Materials and Methods: In the present study, we show that FASN inactivation by specific small interfering RNA (siRNA) promoted the downregulation of the S-phase kinase associated-protein kinase 2 (SKP2) and the consequent induction of p27^KIP1 in HCC cell lines. Results: Expression levels of FASN and SKP2 directly correlated in human HCC specimens and predicted a dismal outcome. In addition, forced overexpression of SKP2 rendered HCC cells resistant to the treatment with the FASN inhibitor C75. Furthermore, FASN deletion was paralleled by SKP2 downregulation and p27^KIP1 induction in the AKT-driven HCC preclinical mouse model. Moreover, forced overexpression of an SKP2 dominant negative form or a p27^KIP1 non-phosphorylatable (p27^KIP1-T187A) construct completely abolished AKT-dependent hepatocarcinogenesis in vitro and in vivo. Conclusions: In conclusion, the present data indicate that SKP2 is a critical downstream effector of FASN and AKT-dependent hepatocarcinogenesis in liver cancer, envisaging the possibility of effectively targeting FASN-positive liver tumors with SKP2 inhibitors or p27^KIP1 activators. Full article

Apolipoprotein E (ApoE) is a lipid carrier in both the peripheral and the central nervous systems (CNSs). Lipid-loaded ApoE lipoprotein particles bind to several cell surface receptors to support membrane homeostasis and brain injury repair. In the brain, ApoE is produced predominantly by astrocytes, but it is also abundantly expressed in most neurons of the CNS. In this study, we addressed the role of ApoE in the hippocampus in mice, focusing on its role in response to radiation injury. To this aim, 8-week-old, wild-type, and ApoE-deficient (ApoE^−/−) female mice were acutely whole-body irradiated with 3 Gy of X-rays (0.89 Gy/min), then sacrificed 150 days post-irradiation. In addition, age-matching ApoE^−/− females were chronically whole-body irradiated (20 mGy/d, cumulative dose of 3 Gy) for 150 days at the low dose-rate facility at the Institute of Environmental Sciences (IES), Rokkasho, Japan. To seek for ApoE-dependent modification during lineage progression from neural stem cells to neurons, we have evaluated the cellular composition of the dentate gyrus in unexposed and irradiated mice using stage-specific markers of adult neurogenesis. Our findings indicate that ApoE genetic inactivation markedly perturbs adult hippocampal neurogenesis in unexposed and irradiated mice. The effect of ApoE inactivation on the expression of a panel of miRNAs with an established role in hippocampal neurogenesis, as well as its transcriptional consequences in their target genes regulating neurogenic program, have also been analyzed. Our data show that the absence of ApoE^−/− also influences synaptic functionality and integration by interfering with the regulation of mir-34a, mir-29b, and mir-128b, leading to the downregulation of synaptic markers PSD95 and synaptophysin mRNA. Finally, compared to acute irradiation, chronic exposure of ApoE null mice yields fewer consequences except for the increased microglia-mediated neuroinflammation. Exploring the function of ApoE in the hippocampus could have implications for developing therapeutic approaches to alleviate radiation-induced brain injury. Full article

This study aimed to investigate the impact of cold stress and priming on photosynthesis in the early development of maize and soybean, crops with diverse photosynthetic pathways. The main objectives were to determine the effect of cold stress on chlorophyll a fluorescence parameters and spectral reflectance indices, to determine the effect of cold stress priming and possible stress memory and to determine the relationship between different parameters used in determining the stress response. Fourteen maize inbred lines and twelve soybean cultivars were subjected to control, cold stress, and priming followed by cold stress in a walk-in growth chamber. Measurements were conducted using a portable fluorometer and a handheld reflectance instrument. Cold stress induced an overall downregulation of PSII-related specific energy fluxes and efficiencies, the inactivation of RCs resulting in higher energy dissipation, and electron transport chain impairment in both crops. Spectral reflectance indices suggested cold stress resulted in pigment differences between crops. The effect of priming was more pronounced in maize than in soybean with mostly a cumulatively negative effect. However, priming stabilized the electron trapping efficiency and upregulated the electron transfer system in maize, indicating an adaptive response. Overall, this comprehensive analysis provides insights into the complex physiological responses of maize and soybean to cold stress, emphasizing the need for further genotype-specific cold stress response and priming effect research. Full article

Background: In 2016, the Global Polio Eradication Initiative (GPEI) recommended the cessation of using type 2 oral poliovirus vaccine (OPV) and OPV, with countries having to switch from the trivalent to bivalent OPV (bOPV) with the addition of inactivated poliovirus vaccine (IPV) in their routine immunization schedule. The current GPEI strategy 2022–2026 includes a bOPV cessation plan and a switch to IPV alone or a combination of vaccine schedules in the future. The focus of our study was to evaluate the immunogenicity of monovalent OPV type 1 (mOPV1) with IPV and IPV-only schedules. Methods: This was a three-arm, multi-center randomized–controlled trial conducted in 2016–2017 in India. Participants, at birth, were randomly assigned to the bOPV-IPV (Arm A) or mOPV1-IPV (Arm B) or IPV (Arm C) schedules. Serum specimens collected at birth and at 14, 18, and 22 weeks old were analyzed with a standard microneutralization assay for all the three poliovirus serotypes. Results: The results of 598 participants were analyzed. The type 1 cumulative seroconversion rates four weeks after the completion of the schedule at 18 weeks were 99.5% (97.0–99.9), 100.0% (97.9–100.0), and 96.0% (92.0–98.1) in Arms A (4bOPV + IPV), B (4mOPV1 + IPV), and C (3IPV), respectively. Type 2 and type 3 seroconversions at 18 weeks were 80.0% (73.7–85.1), 76.9% (70.3–82.4); 93.2% (88.5–96.1), 100.0% (98.0–100.0); and 81.9% (75.6–86.8), 99.4% (96.9–99.9), respectively, in the three arms. Conclusions: This study shows the high efficacy of different polio vaccines for serotype 1 in all three schedules. The type 1 seroconversion rate of mOPV1 is non-inferior to bOPV. All the vaccines provide high type-specific immunogenicity. The program can adopt the use of different vaccines or schedules depending on the epidemiology from time to time. Full article

Introduction: Soluble guanylyl cyclase (sGC) activation in vascular smooth muscle has the potential to induce vasodilation. Chronic sGC activation enhanced vascular function in the congestive heart failure animal model. Therefore, sGC activation can lead to vasodilation and improvement in endothelial function. Objective: To investigate whether the selective sGC activator can revert the endothelial dysfunction and investigate the mechanism of action. Methods: Wistar rats were split into two groups: normotensive (2K) and hypertensive rats (2K-1C). Intact aortic rings were placed in a myograph and incubated with 0.1 µM ataciguat for 30 min. Cumulative concentration-effect curves were generated for acetylcholine (Ach) to assess endothelial function. The pD2 and maximum relaxant effect (Emax) were measured to Ach. In endothelial cell culture, superoxide anion (O₂^•−) was detected by using fluorescent probes, including DHE and lucigenin. Results: Ataciguat improved the relaxation induced by acetylcholine in 2K-1C (pD2: 6.99 ± 0.08, n = 6) compared to the control (pD2: 6.43 ± 0.07, n = 6, p < 0.05). The aortic rings were also improved from 2K (pD2: 7.04 ± 0.13, n = 6) compared to the control (pD2: 6.59 ± 0.07, n = 6, p < 0.05). Moreover, Emax was improved by ataciguat treatment in the 2K-1C aorta (Emax: 81.0 ± 1.0; n = 6), and 2K aorta (Emax: 92.98 ± 1.83; n = 6), compared to the control (Emax 2K-1C: 52.14 ± 2.16, n = 6; and Emax 2K: 76.07 ± 4.35, n = 6, p < 0.05). In endothelial cell culture, treatment with ataciguat (0.1, 1, and 10 µM) resulted in a reduction of the superoxide anion formation induced by angiotensin II. Conclusions: Our findings indicated that ataciguat effectively enhanced endothelial function through the inactivation of superoxide anions. Full article

The effectiveness of COVID-19 vaccines developed against the original virus strain deteriorated noticeably in efficacy against the Omicron variant (B.1.1.529). Moreover, the immunity developed after vaccination or due to natural infection rapidly waned. In the present study, covering this period, we summarize the incidence of breakthrough infections among healthcare workers (HCWs) with respect to administration of the three vaccine doses. Additionally, we evaluate the long-term SARS-CoV-2-specific humoral and T cell responses at two different time points: six and twelve months after receipt of the third (booster) dose. The spike-protein-specific antibody levels and the quantity of structural-protein-specific T cells were evaluated at these time points and compared with the values measured earlier, 14 days after the booster vaccination. The study participants were categorized into two cohorts: Members of the first cohort received a two-dose BNT162b2 mRNA-based vaccine regimen, followed by an additional BNT162b2 booster six months later. Individuals in the second cohort received an inactivated-virus-based BBIBP-CorV booster six months after the initial two-dose BNT162b2 vaccination. Overall, 64.3% of participants were infected with SARS-CoV-2 confirmed by PCR or antigen test; however, additional subjects from the first cohort (23%) who did not know about their previous infection but had an anti-nucleocapsid T cell response were also considered virus-experienced. According to our results, no statistically significant difference was found between the two cohorts regarding the SARS-CoV-2-specific T cell response, neutralizing anti-RBD IgG, and anti-S IgA serum antibody levels either six or twelve months after receiving the booster, despite the overall higher median values of the first cohort. The only significant difference was the higher anti-S1/S2 IgG antibody level in the first cohort one year after the BNT162b2 booster (p = 0.039). In summary, the BNT162b2 and BBIBP-CorV boosters maintain durable humoral and T cell-mediated immune memory even one year after application. Although the booster provided limited protection against Omicron breakthrough infections, as 73.6% of these infections occurred after the booster vaccination, which means 53.5% cumulative incidence, it still offered excellent protection against severe disease and hospitalization in both cohorts. Full article