Search Results (19,939)

The CRISPR/Cas9 system has been widely used for gene editing in various species; however, mosaicism remains a significant challenge. This study aimed to improve gene editing efficiency and reduce mosaicism in porcine embryos by exploring double electroporation pre- and post-in vitro fertilization combined with zona pellucida (ZP) removal. We evaluated the effects of these treatments on the development and mutation rates of oocytes/zygotes edited with guide RNAs (gRNAs) targeting GGTA1, CMAH, or B4GALNT2 genes. Double electroporation significantly increased the total and biallelic mutation rates in ZP-intact zygotes but not in ZP-free zygotes edited using GGTA1-targeted gRNAs. All blastocysts from ZP-free zygotes exhibited biallelic mutations following double electroporation. For the CMAH gene, all blastocysts exhibited mutations (biallelic mutations ≥ 80%); however, double electroporation and ZP removal did not affect their mutation rates or efficiency. For the B4GALNT2 gene, double electroporation significantly increased total mutation rates in ZP-intact zygotes, whereas all blastocysts from ZP-free zygotes showed biallelic mutation. These findings suggest that double electroporation, particularly with ZP removal, may enhance gene-editing efficiency, reduce mosaicism and improve the success of genetic modifications. Full article

Background/Objectives: In February 2024, a nationwide medical crisis in South Korea caused a massive withdrawal of resident physicians. We described changes in patient characteristics and early clinical outcomes among emergency department (ED)-admitted inpatients during this disruption. Methods: We retrospectively analyzed 8149 internal medicine admissions via the ED at a tertiary hospital, 6 months pre- and post-crisis. Multivariable logistic regression evaluated early clinical outcomes, adjusting for baseline confounders. Results: Post-crisis, the internal medicine physician workforce decreased by 36%. Total admissions dropped, while patient acuity increased. After adjustment, the post-crisis group exhibited higher odds of Orders for Life-Sustaining Treatment documentation (adjusted odds ratio [aOR] 1.53, 95% confidence interval [CI] 1.34–1.74), inter-hospital transfers (aOR 1.71, 95% CI 1.49–1.96), and 48 h mortality (aOR 1.81, 95% CI 1.25–2.61). However, adjusted overall in-hospital mortality did not significantly differ (aOR 1.10, 95% CI 0.95–1.27). Conclusions: The crisis led to decreased admissions and higher patient acuity. Despite these shifts, adjusted in-hospital mortality did not significantly differ. This suggests that during severe workforce shortages, acute care was concentrated on a highly selected, high-acuity patient cohort, accompanied by an increased reliance on inter-hospital transfers. Full article

The biosynthesis of sex pheromones in lepidopteran pheromone glands is tightly regulated by pheromone biosynthesis-activating neuropeptide (PBAN) signaling; yet the contribution of non-coding RNA-mediated post-transcriptional regulation remains largely unclear. This study aimed to characterize temporal transcriptomic changes, candidate non-coding RNA-mediated regulatory associations, and temporal molecular dynamics underlying transcriptional remodeling after PBAN treatment in Ostrinia furnacalis. First, we performed comprehensive whole-transcriptome sequencing (WTS) on 18 biologically independent samples collected at six time points (0, 20, 40, 60, 90, and 120 min) after PBAN injection. Then, we systematically identified and quantified the dynamic expression patterns of differentially expressed (DE) mRNAs, miRNAs, lncRNAs, and circRNAs in response to PBAN stimulation. By integratively analyzing these multidimensional omics datasets and inferring sequence-based interaction relationships, we inferred a dynamic candidate competing endogenous RNA (ceRNA) like regulatory network. The candidate ceRNA network anchored four core node genes: the PBAN receptor (PBANR), the rate-limiting enzyme acetyl-CoA carboxylase (ACC), and the terminal biosynthetic enzymes desaturase (DES) and fatty acyl-CoA reductase (FAR). The qRT-PCR results further support the temporal expression pattern of key genes during the PBAN response, suggesting that this network can provide a valuable resource for further functional studies. Full article

Background: Abnormalities in energy and amino acid metabolism are potentially involved in hepatocellular carcinoma (HCC) development. This study aimed to identify serum metabolites predictive of HCC following sustained virological response (SVR) with hepatitis C virus (HCV) treatment. Methods: Comparative metabolomics was conducted using time-course serum samples from patients who failed interferon-based therapy but subsequently achieved SVR with direct-acting antivirals (DAAs), minimizing inter-individual variability. Predictive biomarkers for post-SVR HCC were extracted from the results and validated by comparing 29 patients who developed post-SVR HCC with 58 age-matched patients who remained HCC-free during follow-up. Results: Metabolite concentrations changed more markedly after treatment in SVR cases than in non-SVR cases. Significant changes in methionine (Met), methionine sulfoxide (MetO), and ornithine (Orn) levels before and after treatment (Pre- and Post-Tx) were found only in the non-HCC group. Regression and survival analyses identified high levels of Pre- and Post-Tx Orn, Pre-Tx Met, and Post-Tx MetO as predictors of post-SVR HCC and enabled risk stratification. The integration of these metabolites with the fibrosis-4 (FIB-4) index and alpha-fetoprotein (AFP) facilitated risk stratification and discriminated between high- and low-risk patients. The Pre-Tx FIB-4/Met model and the Post-Tx AFP/MetO/Orn model identified low- and high-risk groups with 3-year HCC incidence rates of 6.4% and 81.8%, respectively. Conclusions: Serum Met, MetO, and Orn were identified as candidate biomarkers associated with post-SVR HCC development, which remains a concern in the fight against hepatitis C. Combining these metabolites with established clinical markers may improve post-SVR HCC risk stratification. Full article

Background/Objectives: Medication-related harm is a leading global patient safety challenge, yet patients’ lived experiences of medication safety remain underexplored in Central and Eastern European healthcare systems, where structural constraints significantly shape everyday medication use. Methods: This study provides an in-depth qualitative analysis of Hungarian patients’ online narratives, building on a prior netnographic mixed-methods study. Using grounded theory-informed principles and a patient-centred medication safety framework, we inductively analysed 5174 publicly accessible Hungarian-language comments posted on health forums and social media platforms between August 2020 and August 2023. The COM-B model was applied as a secondary lens to map findings onto modifiable behavioural determinants. Results: Access to services and communication emerged as the dominant medication safety concerns. Patients reported long waiting times, limited rural emergency services, and brief consultations leading to delayed or inadequate treatment. Communication gaps included insufficient information on medication duration, side effects, and follow-up, as well as conflicting advice from multiple sources, all of which eroded trust and prompted treatment discontinuation or reliance on informal online communities. Community pharmacists were largely absent from patients’ mental models of care, representing a significant missed opportunity given their accessibility. Less frequently mentioned were medication shortages, healthcare professional workload, and systemic safety culture. Conclusions: Clear, respectful communication and timely access to care are central to medication safety from the patient perspective. Netnography combined with a grounded theory-informed methodology offers a valuable approach for capturing authentic patient perspectives in structurally burdened healthcare systems, with findings relevant beyond the Hungarian context. Full article

Background: Endothelial dysfunction and renal injury are emerging as a common feature of long COVID, especially in those with hypertension. It is not yet well characterised whether SARS-CoV-2 infection exacerbates podocyte dysfunction, fibrotic signalling and renal hemodynamic remodelling, over and above the effects of hypertension alone and there are no reliable early biomarkers in this population. Methods: We conducted a comparative cross-sectional study with prospective 6-month treatment response follow-up in 120 adult patients (aged 30–60 years) with essential hypertension (Stage I, II or III; n = 40 per stage), at Bukhara Regional Multidisciplinary Hospital. Each stage subgroup was further divided into post-COVID (3–6 months after recovery; n = 20) and non-COVID (n = 20) strata. Patients with diabetes, known chronic kidney disease, previous myocardial infarction or stroke and other major comorbidities were excluded. Serum cystatin-C, creatinine, aldosterone, TGF-β1 and VEGF-A; urinary nephrin and microalbumin; cystatin-C-derived eGFR (CKD-EPI) and oral protein-loaded renal functional reserve (RFR); and renal Doppler indices (Vps, Ved, RI, PI) of the main, segmental and interlobar arteries were assessed before and after 6 months of guideline-based renin–angiotensin–aldosterone system (RAAS) blockade (enalapril 5–10 mg or azilsartan 40–80 mg, ±eplerenone). Comparisons were made by Student’s t-test—associations by Pearson correlation. Results: At baseline, post-COVID hypertensive patients exhibited consistently higher endothelial–podocyte injury markers than non-COVID counterparts. Urinary nephrin was elevated across all stages (Stage I: 126.5 ± 9.1 vs. 91.9 ± 8.3 pg/mL, p < 0.01; Stage III: 203.3 ± 11.2 vs. 164.5 ± 9.7 pg/mL, p < 0.05), as were VEGF-A (Stage III: 286.1 ± 16.4 vs. 223.2 ± 12.6 pg/mL, p < 0.01) and TGF-β1 (Stage III: 186.4 ± 10.1 pg/mL, 1.3-fold higher; p < 0.01). The detection of microalbuminuria was 100% in Stage III post-COVID patients and 85% in non-COVID controls. The post-COVID groups had selective loss of renal functional reserve (7.8 ± 1.1% in Stage III compared to 12.5 ± 1.6% in non-COVID controls, p < 0.001). Nephrinuria correlated strongly with RFR (r = −0.824, p < 0.001), eGFR (r = −0.797, p < 0.001) and aldosterone (r = 0.613, p < 0.001). Six months of RAAS blockade reduced nephrinuria, microalbuminuria and TGF-β1 in both arms but the magnitude of biomarker reduction appeared smaller in the post-COVID group, particularly in Stage III. Conclusions: Long COVID appears to be associated with persistent endothelial dysfunction and podocyte injury in hypertensive patients. These results indicate that nephrinuria, VEGF-A, TGF-β1 and renal functional reserve are potential exploratory markers of endothelial and renal abnormalities in hypertensive patients following COVID-19. Before clinical utility can be determined, larger studies with multivariable modelling, diagnostic-performance analyses and correction for multiple testing are needed. The differences in biomarker response between groups observed in this study need to be confirmed in larger prospective studies with multivariable modelling and formal interaction analyses. Full article

Background/Objectives: Verb anomia is a common symptom of primary progressive aphasia (PPA), particularly in the logopenic variant (lvPPA). Despite the central role of verbs in sentence construction, interventions specifically targeting verb anomia remain scarce. This study examined the effects of Personalized Observation, Execution and Mental Imagery (POEM) therapy, grounded in evidence that sensorimotor systems are recruited during action verb processing and may support verb retrieval. Methods: A 74-year-old woman with lvPPA completed pre- and post-POEM assessments (linguistic, cognitive, thymic, and motor). POEM consisted of 15 telepractice sessions delivered three times weekly using a systematic procedure with three sensorimotor strategies: action observation, gesture execution, and mental imagery. Results: No significant gains were observed in verb production across naming tasks, spontaneous speech, or functional communication contexts. However, co-verbal gesture use increased in frequency and quality, particularly during the naming of untrained verbs, suggesting that the gesture execution strategy was generalized. A delayed treatment effect was also noted, raising questions regarding the optimal duration of POEM in neurodegenerative conditions. Conclusions: While no statistically significant improvements in verb production were observed, qualitative analyses revealed increased use and quality of co-verbal gestures, suggesting that POEM had a compensatory effect. Future research involving larger cohorts and longer periods could help clarify the benefits of POEM therapy. It would also be relevant to compare in-person and remote delivery formats to evaluate POEM therapy’s robustness and adaptability in different clinical contexts. To conclude, these findings remain preliminary and should be interpreted with caution, particularly given the lack of significant improvement in primary verb production outcomes. Full article

Background and Clinical Significance: Inflammatory myopericardial syndrome is an umbrella term recently introduced by the European Society of Cardiology, which encapsulates the overlap that exists in clinical practice between myocardial and pericardial disease. It has a heterogeneous aetiology and a broad spectrum of severity in terms of its clinical features. Toxoplasma gondii is a rare but recognised infectious cause of myopericarditis and is typically seen in immunocompromised individuals. Case Presentation: We present the case of a young, immunocompetent male, presenting with pleuritic chest pain following a recent flu-like illness. Investigations revealed an acute myocardial injury based on elevated troponin T levels, in the absence of ventricular dysfunction. Toxoplasma immunoserology was consistent with primary toxoplasma infection. The remainder of his viral panel was negative. There was prompt symptom improvement following commencement of treatment with colchicine and a non-steroidal anti-inflammatory agent. Cardiac magnetic resonance imaging post-discharge revealed findings consistent with prior myocarditis. Conclusions: This case is an example of the rare occurrence of toxoplasma myopericarditis in an immunocompetent individual. Cardiac MRI is an invaluable imaging modality used to evaluate myocardial function and tissue characteristics in patients presenting with inflammatory myopericardial syndrome. Full article

This study evaluates regulated deficit irrigation (IR) and post-harvest dehydration (DH) as complementary strategies to mitigate extreme thermal stress on the red grape variety Avgoustiatis during the hot 2024 vintage. Analysis of the berries reveals that while IR significantly expanded vine productivity to 2.75 kg/vine compared to 1.32 kg/vine recorded in control vines (CO), it successfully maintained berry weight (240 g). Conversely, DH induced controlled water loss, reducing berry weight to 93 g and concentrating must sugars to 27.3 ^°Brix, relative to the 23.2 ^°Brix observed in IR. Crucially, both IR and DH prevented the thermal degradation of total acidity (6.73 g/L and 7.25 g/L respectively) which caused by heat stress in CO samples (6.21 g/L). In the finished wines, both practices increased colour intensity by lowering anthocyanin extractability. However, chemical profiling clearly differentiated the treatments with DH maximized skin tannins (164.7 mg/L), yielding highly structured, astringent wines characterized by plum aromas driven by elevated nerol content (492.91 μg/L). Conversely, IR wines presented a more complex volatile profile, boosting fruity and floral notes. In conclusion, as irrigation becomes increasingly restricted by water scarcity under climate change, post-harvest dehydration offers an effective alternative for producing premium, structurally dense red wines. Full article

In this study, charge-trapping memory (CTM) transistors were developed using indium gallium zinc oxide (IGZO) as the oxide semiconductor channel and high-k HfO₂ as the charge-trapping layer, aiming for next-generation nonvolatile memory applications. To evaluate the impact of plasma exposure on film quality and device performance, HfO₂ thin films were deposited via atomic layer deposition (ALD) using both direct plasma (DP) and remote plasma (RP) modes. Post-deposition annealing (PDA) was applied to the IGZO and HfO₂ layers, with experiments conducted at various annealing temperatures to enhance the interfacial stability between the HfO₂ layer and the IGZO channel. Electrical characterization results demonstrated that the RP-processed devices exhibited a wider memory window, reduced gate leakage current, and improved threshold voltage stability compared with the DP-processed devices. Thermal treatment effectively reduced the interfacial defect density and enhanced the crystallinity at the dielectric–channel interface. These findings underscore that the selection of the plasma process and annealing conditions is critical in determining the electrical characteristics and reliability of oxide semiconductor-based CTM devices. Full article

Trichosporon is a type of non-candida yeast-like fungus. At one time, it was commonly reported in immunocompromised patients, but after the introduction of fluconazole as prophylaxis and for the treatment of fungal infections, there was a decrease in the incidence of the disease. With the introduction of echinocandins as the first line of treatment for fungal infections, and the intrinsic resistance of Trichosporon to the drug, there has been a small but increased reported incidence of the disease. Trichosporon usually causes skin infections, but invasive disease can occur in vulnerable patients. Endocarditis due to Trichosporon has been reported rarely, and usually occurs in prosthetic valves. In this paper, we report a patient with dialysis-dependent chronic kidney disease who presented with fever and was found to have native aortic valve endocarditis. In view of the large vegetation, he underwent early aortic valve replacement. Both the blood and tissue cultures grew Trichosporon spp. Post-operatively, he developed fungal septic shock, deteriorated, and died. Invasive Trichosporon disease has been associated with high rates of mortality ranging from 30 to 90%. There is limited literature on endocarditis resulting from Trichosporon. Specific treatment recommendations are unavailable, and a combination of surgery and prolonged antifungal medication will generally be required. Full article

Metanephrine (MN) and normetanephrine (NMN) are critical biomarkers for neuroendocrine tumors (pheochromocytoma and paraganglioma). Following our previous development of a molecularly imprinted solid-phase extraction (MISPE) sorbent for urine analysis, this study evaluated MISPE coupled with HILIC-MS/MS for determining metanephrines in human plasma. Unlike conventional phases, the novel polymer selectively binds analytes as in situ-generated anions via quaternary alkylammonium groups in hydroxide form, ensuring accurate extraction from just 25 µL of plasma. Validated per U.S. FDA guidelines, the assay showed good intra- and interday precision (CV < 10.8%), accuracy (bias < −10.6%) and excellent linearity (R² > 0.99) across pathological ranges (184.3–877.8 ng/L for MN; 174.8–923.0 ng/L for NMN), with low relative standard errors (<6.9%). Excellent selectivity was demonstrated in the presence of structurally close analogs (catecholamines, DOPA and its derivatives). Compared with commercial WCX, the sorbent yielded cleaner extracts, significantly reducing the phospholipid interference. Although lower limits of quantification (92.2 ng/L MN; 87.4 ng/L NMN) slightly exceeded healthy upper thresholds, the method has potential for use in specific clinical scenarios with pronounced biomarker elevations: diagnosis of pheochromocytoma/paraganglioma, monitoring post-treatment metanephrine decline, and tracking tumor-induced hypertensive crises in emergencies. This accessible protocol forms a solid foundation for advanced diagnostics. Full article

Background: The objective was to synthesize and critically appraise systematic reviews with meta-analysis evaluating the association between irrigation, instrumentation, and obturation procedures and post-operative endodontic pain. Methods: An umbrella review was conducted following PRISMA guidelines. Electronic searches identified systematic reviews published between 2016 and 2025. Eligible studies are systematic reviews that include meta-analyses, published in English and correlating the presence of post-operative pain in 3 different critical stages of root canal treatments, namely irrigation, instrumentation and obturation. Methodological quality was assessed using the AMSTAR 2 tool. Outcomes included pain prevalence and intensity at different time points. Results: Out of 368 records, 25 systematic reviews with meta-analysis met the inclusion criteria: 9 on irrigation, 8 on instrumentation, and 8 on obturation. NaOCl concentrations, irrigant activation, and intracanal cryotherapy were repeatedly reported as being associated with reduced short-term post-operative pain. For instrumentation, most reviews reported lower pain with rotary systems, but two studies found no difference or favored reciprocating kinematics. Apical patency did not appear to increase pain and foraminal enlargement may increase early pain. No clinically consistent differences were observed between bioceramic/calcium silicate-based and resin-based sealers, although calcium silicate sealers seem to support periapical healing. However, the certainty of these findings was limited by heterogeneity, methodological weaknesses, and overlap among primary studies. Methodological limitations were identified across reviews, mainly related to no protocol registration (n = 4), incomplete reporting of excluded studies with justification (n = 11), limited assessment of publication bias, and poor reporting of funding sources for primary studies. Conclusions: Based on current evidence, irrigation, instrumentation, and obturation procedures may influence short-term post-operative pain. However, these findings remain tentative because of heterogeneity, methodological weaknesses, variable review quality, and overlap among primary studies. Further high-quality reviews and clinical trials are needed. Full article

Watercress (Nasturtium officinale R.Br.) is a cruciferous vegetable rich in bioactive compounds that may improve cardiovascular disease (CVD) risk factors. However, clinical evidence regarding its direct impact on CVD risk remains limited. This study evaluated the efficacy of watercress consumption on cardiovascular parameters, oxidative stress biomarkers, and estimated CVD risk in middle-aged Thai adults with low-to-moderate risk. Twenty-five participants completed the randomized, placebo-controlled pilot study. The watercress group (n = 12) consumed 16 dried watercress capsules daily for four weeks, while the placebo group (n = 13) received a placebo. Physical examinations, arterial stiffness, lipid profiles, and biochemical biomarkers were analyzed at baseline and the end of treatment. The 10-year CVD risk was estimated using the Framingham equation. Following the intervention, the watercress group showed significant reductions in both systolic and diastolic blood pressure compared to the placebo group. Within the watercress group, significant improvements from baseline to post-intervention were observed in oxidized LDL, antioxidant capacity (ABTS), and estimated 10-year CVD risk score. However, these three parameters did not reach statistical significance when compared to the placebo group. In conclusion, daily watercress consumption significantly lowers blood pressure and demonstrates a potential dietary option for supporting cardiovascular health. Nevertheless, larger and longer-term clinical trials remain necessary. Full article

Background: Actinic keratosis (AK) is a common premalignant skin disorder associated with chronic ultraviolet exposure and a recognized risk of progression to cutaneous squamous cell carcinoma. Tirbanibulin 1% ointment is an effective short-course field therapy for AK, but its efficacy in hyperkeratotic lesions (Olsen grade II–III) may be limited by reduced drug penetration through a thickened stratum corneum. Keratolytic pretreatment may represent a plausible strategy to improve topical drug delivery in these more challenging lesions. Methods: This retrospective chart review included consecutive adults with Olsen grade II–III AK treated in routine clinical practice with either a bland emollient lead-in followed by tirbanibulin (Group A) or salicylic acid 30% ointment pre-treatment (Decapan, Sanitpharma; Milan, Italy) followed by tirbanibulin (Group B). No study-driven procedures or additional visits were implemented. The 14-day bland emollient lead-in used in Group A was part of the routine clinical management applied during the relevant treatment period and was not introduced or retrospectively constructed for the purposes of the present comparative analysis. Outcomes were extracted from de-identified medical records and photographic documentation obtained as part of standard care. For the purposes of analysis, post-treatment evaluations were grouped into predefined windows of 3–6 weeks (T1), 10–14 weeks (T2), and 22–30 weeks (T3), corresponding approximately to 1, 3, and 6 months after treatment initiation. The primary efficacy endpoints were the Actinic Keratosis Area and Severity Index (AKASI) and Total Lesion Count (TLC). Secondary endpoints included quality of life assessed by the Dermatology Life Quality Index (DLQI). Results: Both treatment regimens were associated with clinically meaningful improvements in AK severity. At T3, mean AKASI was significantly lower in Group B than in Group A (0.86 ± 0.38 vs. 1.35 ± 0.27; p < 0.001), corresponding to reductions from baseline of 60.6% and 36.9%, respectively. Similarly, mean TLC at T3 was significantly lower in Group B than in Group A (4.80 ± 1.5 vs. 6.35 ± 1.6; p < 0.001), corresponding to reductions from baseline of 46.7% and 27.0%, respectively. Quality-of-life outcomes also favored the sequential approach, with lower DLQI scores at T3 in Group B compared with Group A (2.9 ± 1.6 vs. 3.8 ± 1.9; p = 0.006). Both treatments were generally well tolerated. Although the incidence of local skin reactions (LSRs) was similar between groups, Group B showed lower retrospectively documented composite LSR scores and lower patient-reported discomfort (p < 0.001) and lower patient-reported discomfort (p < 0.001). Conclusions: Sequential keratolytic pretreatment followed by tirbanibulin was associated with greater reductions in disease burden and with lower severity of treatment-related local reactions in this retrospective cohort (Olsen grade II–III). This retrospective study suggests that keratolytic pretreatment may represent a useful adjunctive strategy in hyperkeratotic AK treated with tirbanibulin. Prospective randomized studies are warranted to confirm these findings and to define standardized treatment protocols. Full article