Search Results (212)

Background and purpose: The role of adjuvant radiation therapy (RT) in early-stage HER2-positive breast cancer treated with breast-conserving surgery (BCS) and systemic therapy remains uncertain in the era of HER2-targeted regimens. This study evaluates the survival impact of RT in patients aligned with the HERO RT de-escalation trial (NRG-BR008). Materials and methods: We queried the National Cancer Database for patients with early-stage HER2-positive invasive breast carcinoma treated with BCS and systemic therapy, stratified into HERO trial-aligned cohorts: Arm 1 (adjuvant systemic therapy) vs. Arm 2 (neoadjuvant systemic therapy, pathologic complete response). Within each cohort, patients receiving adjuvant RT were compared with those omitting RT. In the primary analysis, patients were propensity score matched (PSM) on demographics, diagnosis years, tumor characteristics, and trial stratification variables. Inverse probability of treatment weighting (IPTW) was additionally performed as a sensitivity analysis. Overall survival was evaluated using Kaplan–Meier, Cox regression, and restricted mean survival time (RMST). Results: In Arm 1 (818 patients, 94 deaths), 5-year OS was 96.9% with RT vs. 88.0% without RT, and 10-year OS was 94.3% vs. 68.5% (log-rank p < 0.001). RT omission was associated with higher mortality in the PSM Cox model (HR, 4.78; 95% CI, 2.84–8.02; p < 0.001), with an RMST advantage favoring RT of +2.86 months at 5 years and +12.55 months at 10 years (p < 0.001). In Arm 2 (176 patients, 10 deaths), 5-year OS was 97.6% with RT vs. 91.1% without RT, and OS at 107 months was 94.8% vs. 91.1% (log-rank p = 0.13). RT omission was not statistically significant in the PSM Cox model (HR, 3.40; 95% CI, 0.82–14.05; p = 0.09), though RMST favored RT (+1.83 months at 5 years, p = 0.004; +3.91 months at 107 months, p = 0.03). IPTW analyses were directionally consistent in Arm 1 (HR, 3.26; 95% CI, 2.52–4.21; p < 0.001) and inconclusive in Arm 2 (HR, 1.78; 95% CI, 0.80–3.95; p = 0.16). Conclusions: In this HERO-aligned national cohort, RT omission was associated with inferior OS in patients treated with adjuvant systemic therapy after BCS. Findings in the neoadjuvant pCR cohort were imprecise and hypothesis-generating. Given the retrospective registry design, lack of recurrence-specific endpoints, and potential residual confounding, results should not be interpreted as causal but support continued RT use outside prospective de-escalation trials. Full article

Background/Objectives: Chemical methods for quantifying resistant starch (RS) in rice are labor-intensive, costly, and lack high repeatability, creating a bottleneck in breeding. This study aimed to develop specific, codominant molecular markers for the Wx gene to enable rapid and accurate genotype screening for RS content, thereby accelerating the development of high-RS rice varieties. Methods: Based on sequence alignment of the Wx gene in rice varieties with divergent RS content, a key single-nucleotide polymorphism was targeted. Two sets of tetra-primer amplification refractory mutation system polymerase chain reaction (ARMS-PCR) markers, T-Wx9-RS1 and T-Wx9-RS2, were designed. These markers were used to genotype diverse rice varieties and F₄ segregating populations, with results validated against standard chemical assays. Results: Sequence analysis identified a critical T → C base mutation at position 202 of the ninth exon in high-RS varieties. The developed ARMS-PCR markers successfully and consistently distinguished all three possible genotypes (homozygous mutant, homozygous wild-type, and heterozygous). The genotyping results showed complete concordance with the phenotypes determined by chemical methods. Conclusions: The developed molecular markers, T-Wx9-RS1 and T-Wx9-RS2, provide a rapid, reliable, and cost-effective tool for marker-assisted selection of high resistant starch content in rice. Their implementation can significantly enhance screening efficiency and expedite the breeding pipeline for novel, nutritionally improved rice cultivars. Full article

Human papillomavirus (HPV) vaccination may eventually eradicate oncogenic vaccine-targeted HPVs but only with a strategy that also protects unvaccinated individuals. We compared the impact of gender-neutral and girls-only vaccination strategies on the indirect and direct protection of unvaccinated and vaccinated young women against HPV16/18 infection using HPV16/18 seropositivity and PCR positivity 3–7 years post vaccination as the outcome measure. A total of 33 Finnish communities were randomized to one of three vaccination strategies: bivalent gender-neutral HPV vaccination (Arm A), girls-only HPV vaccination (Arm B), or control hepatitis B vaccination (Arm C). All individuals born between 1992 and 1995 and residing in these communities (n = 80,272) were invited to participate. Overall, 11,662 males and 20,513 females consented, corresponding to vaccination coverages of 25% and 45%, respectively, in 2007–2009. Between 2010 and 2014, 11,396 cervical samples were collected from 18-year-old participants and subjected to high-throughput PCR-based HPV genotyping. In addition, serum samples were obtained from 8022 unvaccinated women under 23 years of age residing in Arm A (n = 2657), Arm B (n = 2691), or Arm C (n = 2674) communities during the pre-vaccination (2005–2010) and post-vaccination (2011–2016) periods. To assess indirect vaccine effects using PCR and serological outcomes in unvaccinated women, we compared reductions in HPV16/18 prevalence from baseline within the gender-neutral and girls-only vaccination arms, using the control arm as a reference. A significant decrease in seroprevalence between the pre- and post-vaccination periods was detected in the gender-neutral communities for both HPV16 (seroprevalence ratio = 0.64) and HPV18 (0.72), whereas no comparable reductions were observed in the girls-only or control communities. In contrast, a significant reduction in HPV18 PCR-based prevalence from baseline to the post-vaccination period was observed in both the gender-neutral (0.32) and girls-only (0.61) communities. However, after accounting for ratios of seroprevalence rations for secular trends, the corresponding decrease in HPV18 seroprevalence was no longer statistically significant. Vaccine efficacy (VE) in Arm A or Arm B versus Arm C of vaccinated women measured the direct protection of vaccinated women by vaccination strategy. HPV16/18 VEs varied between 89% and 96% with some indication of herd effect against HPV18. Robust effectiveness of vaccination against PCR-confirmed cervical HPV16/18 infections, along with rapid indirect protection against HPV16/18 and HPV18 infections, was evident even with vaccination reaching only 25% and 45% coverage. Our results suggest that vaccine efficacy and herd effect induced by gender-neutral 2vHPV vaccination sets the stage for comprehensive HPV eradication, including the unvaccinated in the vaccinated communities. Full article

Growing evidence has revealed that gut dysbiosis is associated with the development of anxio-depressive disorders through mechanisms that involve neuroimmune signaling, neurotransmitter changes, and neuroplasticity in the brain. This study investigated the effects of gingerol-enriched ginger (GEG) on specifically anxiety-related neuroinflammation-, neuroimmunity-, neuroplasticity-, neurotransmission-, and neurotoxicity-associated genes in different brain regions, as well as on alterations linked to colonic microflora-driven dysbiosis, in the spinal nerve ligation (SNL) rat model of neuropathic pain (NP). Twenty-seven male rats were assigned to 3 groups: sham, SNL, and SNL-treated with GEG at 200 mg/kg body weight (SNL+200GEG) via oral gavage for 5 weeks. Anxiety-like behavior was assessed on the elevated plus maze (EPM). mRNA expression was assessed by qRT-PCR using respective primers. Correlation between behavioral parameters and colonic microbiome composition was analyzed using the Spearman rank correlation. The SNL+200GEG group demonstrated decreased anxiety-like behavior in the SNL model. Compared to the SNL group, the SNL+200GEG group had increased mRNA expression of NRF2 (amygdala: left), LXRα (amygdala: both sides), and CX3CR1 (amygdala: both sides, hippocampus: right). GEG modulated neuroplasticity as shown by increased gene expression of PGK1 (amygdala: right, hippocampus: both sides), MEK1 (frontal cortex: both sides), LDHA (frontal cortex: both sides), GPM6A (frontal cortex: both sides, amygdala: right, hippocampus: right, and hypothalamus), and GLUT1 (amygdala: right) as well by decreased gene expression of HIF1α (in all brain regions except for the hypothalamus). GEG modulated neurotransmission via clearance of excessive glutamate release as suggested by increased gene expression of SLC1A3 (frontal cortex: both sides, hippocampus: right) and via augmenting mGluR5 signaling as shown by increased gene expression of GRM5 (hippocampus: both sides, hypothalamus) as well as downregulation of KMO, HAAO, GRIN2B, and GRIN2C influencing downstream serotonergic neurotransmission and NMDA receptor-mediated glutamatergic pathways in different brain regions. GEG further alleviated neurotoxicity through downregulated gene expression of SIRT1, KMO, IDO1, and HAAO in different brain regions. Moreover, the increased relative abundance of Bilophila spp., accompanied by decreased time spent in the EPM open arms, suggests that increased Bilophila abundance increases anxiety-like behavior. GEG supplementation mitigated anxiety-like behavior in male rats with NP, at least in part, by reducing SNL-induced inflammatory sequelae-related mRNA gene expression in different brain regions. In addition, there is a positive correlation between the abundance of Bilophila wadsworthia and the degree of anxiety-like behavior. Full article

Background: Rapid multiplex PCR assays promise faster and broader detection of uropathogens and resistance markers than conventional quantitative urine culture and susceptibility testing (C&S), but trial evidence linking PCR-guided management to patient-centered outcomes and the mechanisms of any benefit is limited. We performed an ad hoc analysis of the randomized, multicenter NCT06996301 trial to evaluate whether PCR-guided diagnostic management improves clinical symptom resolution in complicated urinary tract infection (cUTI) and to quantify mediation by time-to-antibiotic start and antibiotic appropriateness. Methods: Paired PCR and C&S were collected for all participants; treating investigators received and acted on randomized results from one diagnostic modality and remained blinded to the comparator. The modified intention-to-treat (Mod-ITT) cohort at end-of-study (EOS) included 362 participants (PCR n = 193; C&S n = 169). The primary outcome was complete clinical cure at EOS (absence of all baseline symptoms). Secondary outcomes included partial cure (≥50% symptom reduction) and per-symptom changes. We used mixed-effects logistic regression (site random intercept) to estimate associations, and causal mediation analysis with nonparametric bootstrap (B = 2000) to decompose PCR’s total effect into indirect effects via time-to-antibiotic (log-transformed) and antibiotic appropriateness (binary, adjudicated at EOS) for complete clinical cure and partial cure. Results: Median time-to-first antibiotic was substantially shorter in the PCR arm (20 h; IQR 12–36) than in the C&S arm (52 h; IQR 30–66; p < 0.001). Antibiotic appropriateness was higher after PCR-guided care (161/193; 83.4%) versus C&S (105/169; 62.1%; p < 0.001). Complete clinical cure occurred in 143/193 (74.1%) PCR versus 106/169 (62.7%) C&S (p = 0.020); partial cure in 161/193 (83.4%) versus 121/169 (71.6%; p = 0.014). In a total-effect mixed model (no mediators), PCR assignment was associated with higher odds of cure (adjusted OR 1.95; 95% CI 1.12–3.39; p = 0.018). In the mechanistic model including mediators, antibiotic appropriateness (OR 2.48; 95% CI 1.45–4.24; p = 0.001), and time-to-antibiotic (per 1 h, OR 0.95; 95% CI 0.926–0.975; p < 0.001) were independently predictive, while the direct arm effect was attenuated (OR 1.10; 95% CI 0.33–3.71). Mediation analysis estimated a statistically significant combined indirect effect (ACME) of 0.0648 (95% CI 0.0343–0.0977), ADE 0.0207 (95% CI −0.0282–0.0784), total effect 0.0796 (95% CI 0.0419–0.1225), and proportion mediated ≈ 74%. Both time-to-antibiotic and appropriateness contributed, with ACME_time ≈ 0.046 and ACME_appropriateness ≈ 0.019. Exploratory analysis using partial cure as the outcome confirmed the robustness and internal validity of the complete-cure findings. Conclusions: In this ad hoc analysis of a randomized trial, PCR-guided management of cUTI improved patient-centered symptom outcomes compared with culture-guided care. Most of the benefit was mediated through faster initiation of antibiotics and, to a lesser extent, increased probability of an appropriate initial antibiotic. These results support stewardship-integrated, rapid molecular diagnostics (used alongside culture) to shorten time-to-effective therapy and improve clinical outcomes in cUTI. Full article

Objectives: To investigate the phenotypic and genotypic changes of Acinetobacter baumannii collected from the tertiary oncology setting in Lithuania. Methods:A. baumannii isolates (n = 61) were collected in the years 2013–2014 (n = 28) and 2017–2019 (n = 33) from a tertiary care cancer center in Lithuania. Antimicrobial susceptibility was determined according to EUCAST and for piperacillin/tazobactam and cefepime, according to CLSI guidelines. PCR, pulsed-field gel-electrophoresis, and multi-locus sequence typing were used for resistance gene detection and genotyping. The biofilm formation ability was determined by a microtiter plate assay. Results: Of 61 A. baumannii isolates obtained, 84% (51/61) and 71% (43/61) were multi-(MDR) and extensively (XDR) drug-resistant, respectively. Carbapenem-resistant isolates comprised 77% (47/61); of these, 92% (43/47) harbored genes encoding the OXA-23-like, and 4% (2/47) OXA-24-like carbapenemases. All isolates were susceptible to colistin. Genotyping analysis revealed six groups with the highest prevalence of international clones 1 (IC1) and 2 (IC2), which dominated during 2013–2014 and 2017–2019, respectively. Notably, the A. baumannii diversity increased in 2017–2019 with the emergence of 3-LST groups G4, G8, G12, and G14, which included isolates of ST276, ST78, ST1463, and ST1336 sequence types, respectively. The IC1 and IC2 isolates displayed characteristic gene profiles aacC1, aacC2, aphA6, sul1, and armA, strA-strB, bla_TEM, respectively, whereas isolates from other groups had lesser resistance gene content. Isolates from IC2, G12, and G14 groups were strong biofilm producers; IC1, G4, and G8 isolates displayed no/weak biofilm formation capacity. Conclusions: A. baumannii from the cancer center showed a high prevalence of MDR and XDR phenotypes. Clonal dominance and diversity changed during the surveillance periods with the replacement of IC1 by IC2 clone isolates and the emergence of higher clonal diversity of isolates with stronger biofilm-forming capacity. The observed changes indicate a concerning trend of the establishment of a more virulent A. baumannii in the cancer setting. Full article

Background: Rapid molecular detection of antimicrobial resistance (AMR) can shorten time to effective therapy in complicated urinary tract infections (cUTI), but the ability of gene presence and quantitative PCR signal (Ct, and ΔCt = Ct_marker − IC_Ct) to predict phenotypic non-susceptibility and clinical outcomes requires rigorous evaluation. We analyzed marker-level concordance, Ct→MIC relationships, and the clinical impact pathway in the randomized NCT06996301 trial. Methods: Marker–phenotype concordance metrics (sensitivity, specificity, PPV, NPV, LR+, LR−, κ) were computed for selected marker × species strata with stable sample sizes. Mixed-effects models (log₂[MIC] ~ ΔCt_marker + IC_Ct + collection_method + prior_abx + (1|site)) assessed quantitative Ct→MIC associations. ROC analyses evaluated ΔCt discrimination of phenotypic non-susceptibility. A pre-specified sensitivity analysis included smaller strata (n ≤ 20) with bootstrap 95% confidence intervals for ΔCt slopes and AUCs. Clinical analyses compared PCR-guided (n = 193) versus culture-guided (n = 169) arms for time-to-antibiotic and treatment success using adjusted logistic regression and causal mediation (time-to-antibiotic as mediator; bootstrap inference). Results: High genotype–phenotype concordance was observed for canonical markers (e.g., bla_CTX-M in E. coli: sensitivity 0.94 [95% CI 0.88–0.97], specificity 0.995 [95% CI 0.990–0.998], κ ≈ 0.93). Mixed models showed modest but significant Ct→MIC associations for select markers (e.g., bla_CTX-M in E. coli: ΔCt slope −0.15 [95% CI −0.27 to −0.02], p = 0.015). The sensitivity analysis (n ≤ 20 strata) confirmed consistent negative directions, with robust bootstrap CIs excluding zero for qnrS (E. coli), tetM (E. coli), blaNDM (Klebsiella), and qnrS (Proteus). ROC AUCs for ΔCt prediction of non-susceptibility ranged from 0.62 to 0.81 (95% CIs ≈ 0.47–0.97). Clinically, PCR guidance shortened median time to antibiotic initiation (20 h vs. 52 h) and increased treatment success (88.1% vs. 78.1%; adjusted OR 1.95 [95% CI 1.12–3.40], p = 0.018). Mediation analysis estimated that 63% (ACME 0.112 [95% CI 0.045–0.178], p = 0.002) of the treatment success benefit was mediated through earlier antibiotic initiation. Conclusions: Binary detection of high-impact AMR genes by multiplex PCR reliably predicts phenotypic non-susceptibility and accelerates effective therapy when integrated with stewardship workflows. Quantitative PCR (ΔCt) provides modest but reproducible information about MIC magnitude and may flag heteroresistant subpopulations. A pragmatic implementation model combining rapid PCR with conventional culture is recommended to optimize clinical benefit while retaining isolate recovery for definitive AST. Full article

Vitamin D deficiency is highly prevalent in Pakistan, but there is limited data on its genetic aspects. This case–control pilot study aimed to determine the association of rs782153744, rs200183599, rs118204011, and rs28934604 with vitamin D deficiency along rs7041 which has been studied in our population. The DNA of a total of 600 subjects (300 cases and 300 controls) was extracted and genotyped by tetra ARMS PCR, followed by Sanger DNA sequencing of exon 4 of the CYP2R1 and CYP27B1 genes and exon 8 of the GC gene. SNP Stat was employed to analyze the data, while logistic regression was used to calculate the p-values and odds ratios (ORs). The R package version R studio (2025.05.1) Build 513 was used to statistically analyze rs782153744. In silico modeling of wild and mutant CYP2R1 and GC proteins was performed in Swiss-Model, Swiss-Dock, Discovery Studio, and PyMol using 3c6g and IJ78 as templates to perform binding pocket analysis of vitamin D3. The rs782153744 showed a protective association in the additive (OR: 0.15, 95% CI: 0.08–0.27, p-value < 0.001), recessive (OR: 0.19, 95% CI: 0.10–0.33, p-value < 0.001), and dominant (OR: 0.19, CI = 0.10–0.33, p-value < 0.001) models, while GC rs7041 (T > A, T > G) displayed a p-value < 0.0001 across all genetic models. Sanger sequencing yielded insignificant results, and the SNPs rs200183599, rs118204011, and rs28934604 had no significant association with vitamin D deficiency. The molecular pocket analysis of wild and mutant CYP2R1 proteins carrying rs782153744 polymorphisms revealed no changes. GC proteins carrying the rs7041 polymorphism revealed a shift in their 3D and 2D configuration, as well as a change in the amino acid residue of the binding pocket of VD3. The risk-associated rs7041 and protective rs782153744 variants back genetic screening for vitamin D deficiency risk stratification, allowing targeted supplementation in predisposed subjects and assisting in formulating a genotype-specific therapeutic approach. Full article

Intervertebral disc degeneration is a leading cause of chronic back pain, with existing treatments focusing on symptom management rather than true tissue repair. Cellular therapies—such as platelet-rich plasma (PRP), autologous chondrocytes, and mesenchymal stem cells (MSCs)—have emerged as promising strategies for disc regeneration. In this study, fifteen New Zealand white rabbits underwent fluoroscopy-guided needle puncture of the L4-L5 discs and were allocated to receive PRP alone, PRP-chondrocytes, or PRP-MSCs eight weeks later, while the L3-L4 disc served as a healthy internal control. At 16 weeks post-injury, histological scoring revealed significant improvements in annular integrity, cellularity, and matrix composition in all treated groups compared with untreated lesions, with the greatest gains observed in the PRP-chondrocytes arm, intermediate effects with PRP-MSCs, and more modest changes with PRP alone. Complementary RT-qPCR analysis of COL2A1 and COL10A1 expression confirmed a shift toward a more regenerative phenotype, marked by enhanced COL2A1 and reduced COL10A1 levels, which was most pronounced in the PRP-chondrocytes arm. Despite these advances, none of the interventions fully restored the healthy disc architecture, underscoring the complexity of disc repair. These findings support the potential of combining PRP with chondrocytes or MSCs for intervertebral disc regeneration and demonstrate the need for further optimization of cell doses, PRP formulations, and delivery protocols before clinical translation. Full article

Introduction: Cutaneous leishmaniasis (CL) poses a number of challenges when it comes to diagnosis and treatment, due to the variety of clinical presentations that mimic other conditions and hinder the choice of the most appropriate therapeutic approach, especially in the context of immunodepression. Case presentation: We present the case of a 63-year-old woman on anti-tumor necrosis factor (TNF) therapy, who underwent surgical excision for the diagnostic purposes of a chronic non-healing lesion located on her right arm. The histopathological examination revealed the presence of Leishmania amastigotes. CL relapsed in the following months, with new lesions appearing both close to the excision scar and at a different body site. At this point, in order to avoid another surgical intervention, cutaneous swabs for Leishmania Polymerase Chain Reaction (PCR) were performed on both lesions. Both samples yielded positive results, and the patient was treated with a 4-week course of miltefosine. Conclusions: These results support the use of cutaneous swabs as a highly sensitive and less invasive tool for the diagnostic workup of CL. In addition, our case prompts a reflection on the management of immunosuppressed patients with CL, with particular emphasis on the risk of reactivation or simultaneous involvement of multiple anatomical sites, thus suggesting the need for specific considerations and personalized management for this group of subjects. Full article

Background: Malnutrition is common in chronic hemodialysis (HD) patients and often remains underdiagnosed. While body composition, functional status, and anthropometric measures can support nutritional assessment, their associations with nutritional status are not fully established in this population. This study aimed to evaluate the diagnostic performance of various measures for assessing malnutrition in chronic HD patients, using the Subjective Global Assessment (SGA) as the reference standard. Methods: This cross-sectional study involved chronic HD patients, stratified by nutritional status using the SGA. Data collection consisted of clinical interviews, anthropometric and functional measurements, bioelectrical impedance analysis (BIA), and biochemical analyses. Statistical analysis included Spearman’s correlation, logistic regression, receiver operating characteristic (ROC) curve analysis with area under the curve (AUC) to assess predictive accuracy, standardized effect sizes to show the magnitude of differences, and kappa statistics to evaluate concordance between variables. Results: This study included 103 chronic HD patients. Malnutrition was diagnosed in 50.5% of patients based on the SGA. Phase angle (PA) was the strongest single predictor of malnutrition (AUC = 0.79; specificity 0.88; sensitivity 0.58). PA ≤ 5.1° was significantly associated with higher malnutrition risk (OR: 10.23; 95% CI: 3.93–30.61; p < 0.001). Handgrip strength (HGS) also demonstrated good diagnostic value (AUC = 0.71; specificity 0.84; sensitivity 0.59). A multivariable model incorporating eight parameters—gender, post-dialysis ECW/ICW ratio, post-dialysis lean and fat mass, serum albumin, normalized protein catabolic rate (nPCR), arm circumference (AC), and HGS—achieved an AUC of 0.88 (95% CI: 0.81–0.95) and pseudo-R² of 0.46, demonstrating improved predictive performance. Conclusions: An integrated panel of anthropometric, bioimpedance, functional, and biochemical markers provides superior diagnostic accuracy compared to individual predictors, supporting a holistic diagnostic approach in HD patients. Full article

Background: The efficacy of a single oral dose of Ivermectin as prophylaxis for SARS-CoV-2 is uncertain. This trial sought to evaluate the effectiveness of a single oral low dose of Ivermectin to prevent SARS-CoV-2 infection or reduce symptoms if infection did occur. Methods: Asymptomatic community-dwelling adults were enrolled in this study within 72 h of close contact with a case of SARS-CoV-2. Participants were randomised, stratified by vaccination status and exposure site, to a single oral 200 µg/kg dose of Ivermectin or placebo. The primary outcome was conversion to a positive polymerase chain reaction (PCR) or rapid antigen test (RAT) for SARS-CoV-2 within 14 days of close contact. Secondary outcomes were restricted to those who met the primary outcome. They included the following: days alive free of symptoms in the 14 (DAFS1-14) and 28 (DAFS1-28) days following intervention and days from close contact until a positive PCR or RAT for SARS-CoV-2. Results: A total of 536 participants registered for this trial. Of these, 86 met inclusion criteria and were randomised. 68 adhered to the trial protocol and were included in the analysis. A total of 11/36 (Ivermectin arm) and 11/32 (placebo arm) met the primary outcome. After controlling for age and prior SARS-CoV-2 infection, the estimate (95% confidence interval (95% CI)) of the effect of Ivermectin (compared to placebo) on the absolute value of the proportion of participants converting to a positive PCR or RAT was −0.051 (−0.26 to 0.16), p = 0.63. After controlling for prior SARS-CoV-2 infection, age, body mass index, hypertension and lung disease, the average treatment effect (Ivermectin versus placebo) on DAFS1-14 was 2.5 days (95%CI 1.1 to 4.5), p = 0.036, and for DAFS1-28, was 2.3 days (95% CI 0.7 to 3.3), p = 0.35. The mean (standard deviation) number of days from close contact until a positive PCR or RAT was 5.0 (4.1) days for the Ivermectin group versus 2.6 (0.8) days for the placebo group. After controlling for age and prior SARS-CoV-2 infection, the average treatment effect (95%CI), Ivermectin versus placebo, on days from close contact until a positive PCR or RAT was 2.3 days (95% CI 1.1 to 3.4), p = 0.033. Conclusions: We did not demonstrate that a single oral low dose of Ivermectin administered to asymptomatic adults within 72 h of close contact with a case of SARS-CoV-2 prevents conversion to a positive PCR or RAT. However, the trial had a small sample size and does not exclude a clinically meaningful effect of Ivermectin on conversion to a positive PCR or RAT. Amongst those who did convert to a positive PCR or RAT, the use of Ivermectin significantly lengthened the time from close contact to conversion and increased the number of days alive free of symptoms following intervention. Full article

The mechanisms underlying the abnormal activation of microglia affecting cognitive function under high-altitude hypobaric hypoxia (HAHH) have not been fully elucidated. This study aims to investigate the effects of HAHH on the expression of the receptor for advanced glycation end-products (RAGE) in hippocampal microglia of mice and to explore the role of RAGE inhibitors in alleviating HAHH-induced microglial inflammation and cognitive impairment. Mice were exposed to HAHH via a multi-environment simulation chamber, and RNA sequencing, qPCR, WB, flow cytometry and immunohistochemistry showed that HAHH exposome significantly increased RAGE expression in hippocampal microglia of mice (p < 0.001 vs. normoxia), which was closely related to microglial neuroinflammatory responses. RAGE inhibitor (FPS-ZM1) alleviated HAHH-induced microglial inflammation (TNF-α decreased by 64%, p < 0.001; CD86⁺ cells decreased by 42%, p < 0.001) and improved cognitive function in mice (Y-maze novel arm time: 28.08 ± 5.14 s vs. hypoxia 19.67 ± 4.68 s, p = 0.016; NORT recognition index: 0.52 ± 0.05 vs. hypoxia 0.33 ± 0.07, p < 0.001). Mechanistic studies revealed that RAGE inhibitors reduced microglial inflammation by inhibiting the MAPK pathway and decreasing nuclear translocation of NF-κB p65. Furthermore, high-mobility group box 1 (HMGB1) expression increased under hypoxic conditions (p < 0.001 vs. normoxia) and positively regulated RAGE expression. HMGB1 inhibitors reduced RAGE expression and attenuated HAHH-induced microglial inflammation. Overall, the HAHH exposome induces microglial inflammation via the HMGB1-RAGE-NF-κB pathway. RAGE and HMGB1 inhibitors may serve as novel therapeutic strategies to mitigate HAHH-induced cognitive impairment, providing a theoretical basis for the treatment of cognitive impairment. Full article

Background: Differentiating between bacterial and viral infections in pediatric emergency care is challenging, often leading to unnecessary antibiotic use. The MeMed BV (MMBV) test is a host-response assay designed to differentiate bacterial from viral infections, but real-world data in pediatric settings remain limited. Methods: We conducted a pragmatic, single-center, prospective cohort study to assess the clinical utility of MMBV in children with acute respiratory infections or fever without source. Patients were assigned to standard of care (SOC) or MMBV testing (SOC+MMBV) based on time of presentation to the emergency department. The primary outcome was antibiotic prescribing. Secondary outcomes included diagnostic test utilization, hospitalization rates, and length of stay. Analyses were stratified by hospitalization status, clinical severity [National Institute for Health and Care Excellence (NICE) traffic light system], and patient age. Results: From July 2023 to April 2024, 343 patients were enrolled (171 SOC, 172 SOC+MMBV). In the SOC+MMBV arm, reduced antibiotic prescribing was observed among outpatients and those with non-severe signs and symptoms. Antibiotic prescribing was significantly reduced in children under five years with a low-risk profile, according to the NICE traffic light system (26.3% vs. 7.5%; p = 0.034). Multiplex PCR testing was significantly reduced in the SOC+MMBV group (28.7% vs. 16.3%; p = 0.006) compared to SOC for both inpatients and outpatients. No significant differences were observed in overall diagnostic test use or length of stay. Conclusions: MMBV improved antibiotic and diagnostic stewardship in a real-world pediatric ED setting, significantly reducing unnecessary antibiotic use among low-risk children under five and minimizing unnecessary multiplex PCR testing across the cohort. Full article

Leaf morphology significantly impacts rice (Oryza sativa L.) plant architecture and yield. Here, we identified and characterized a novel narrow-leaf mutant, nal25, derived from indica rice cultivar ‘Huazhan’ using EMS mutagenesis. Phenotypic analyses revealed that nal25 exhibited significantly narrower leaves, reduced plant height, increased tiller number, and notably decreased grain size, seed setting rate, and thousand-grain weight compared to the wild type. Genetic analyses demonstrated that the narrow-leaf phenotype is controlled by a single recessive nuclear gene. Through precise localization analysis, the NAL25 gene was located within a region of approximately 103 kb on the long arm of rice chromosome 7. The sequencing results showed that the mutant nal25 had a T to C mutation at position 173 of the heat-shock protein gene LOC_Os07g09450 encoding the DnaJ domain in this interval, resulting in a change in amino acid 58 from leucine to proline. The qRT-PCR results showed that the expression level of NAL25 gene decreased in the mutant. The nal25 mutant obtained in this study exhibits stable mutant phenotypes, including dwarfism and excessive tillering, traits typically unfavorable for rice production. Nevertheless, it serves as valuable genetic material for forward genetics approaches to identify yield-related genes regulating leaf morphology and culm height. Thus, research on the nal25 mutant advances the development of rice varieties with ideal plant architecture, thereby stabilizing yield increases and safeguarding global food security. Full article