Leber hereditary optic neuropathy (LHON) is the most common mitochondrial disorder, typically causing substantial, often permanent, central vision loss in young adults. It manifests as a subacute optic neuropathy, frequently progressing sequentially in both eyes, due to selective degeneration of retinal ganglion cells (RGCs). The condition is primarily associated with three mitochondrial DNA (mtDNA) point mutations-m.11778G>A, m.14484T>C, and m.3460G>A-located in complex I of the mitochondrial respiratory chain. These mutations impair oxidative phosphorylation, elevate reactive oxygen species (ROS), and trigger apoptosis of RGCs. Although historically considered untreatable, emerging therapies provide new prospects. Idebenone, a synthetic CoQ10 analog, is the first pharmacologic agent approved in Europe, demonstrating partial visual recovery in patients treated early by improving mitochondrial electron transport and reducing oxidative stress. Gene therapy using allotopic expression of ND4 via adeno-associated viral vectors (rAAV2/2-ND4) has shown improvement in both eyes even after unilateral injection. Advanced gene-editing techniques, such as zinc finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs), offer potential approaches for correcting heteroplasmic mutations. High-throughput genetic testing, including whole-genome sequencing and clinical exome analysis, enables precise identification of nuclear modifiers that influence LHON phenotypes, facilitating early diagnosis and intervention. Current clinical trials, including RESTORE and REFLECT, emphasize the importance of prompt treatment to optimize visual outcomes.

Chlamydia psittaci is an obligate intracellular bacterium and an increasingly recognized cause of severe community-acquired pneumonia (CAP) in adults, with contemporary multicenter Chinese cohorts reporting severe-stratum in-hospital mortality clustering around 8-9% and higher figures in acute respiratory distress syndrome (ARDS)-enriched series. β-Lactam therapy lacks reliable activity against this pathogen, yet considerable practice variation persists in the selection, sequencing, and de-escalation of intracellularly active agents in the era of routine metagenomic and targeted next-generation sequencing (mNGS, tNGS). The aim of this review is to provide a phase- and severity-stratified, bedside antimicrobial framework for severe psittacosis in adults-clarifying when to select, continue, switch, or combine intracellularly active agents at the 48- to 72-hour ICU decision points. This narrative review, reported in accordance with the SANRA framework, synthesizes the post-2015 antimicrobial evidence for severe C. psittaci pneumonia in adults and integrates treatment phase (empirical versus targeted), severity context, organ support, and antimicrobial stewardship at the 48- to 72-hour ICU decision points. Drawing on six multicenter cohorts and the largest dedicated multicenter dataset to date (Fang 2026, n = 186; severe-stratum mortality 7/81 = 8.6%), phase-by-severity stratification reconciles apparently discordant tetracycline- and fluoroquinolone-favoring cohort signals. Doxycycline is the preferred targeted backbone in confirmed non-pregnant disease; reflex class-switching is not required when severely ill patients are already improving on an empirical fluoroquinolone at NGS confirmation. Omadacycline is a renal-sparing alternative when acute kidney injury, anticipated continuous renal replacement therapy, or unreliable doxycycline access alters the standard pathway; high-dose tigecycline is reserved for salvage; azithromycin retains its clearest targeted role in pregnancy. Apparent nonresponse at 72 hours should trigger structured reassessment for coinfection, secondary organizing pneumonia, pulmonary embolism, and inadequate antimicrobial exposure before any salvage escalation. Adjunctive corticosteroid use should follow contemporary severe-CAP guidance pending pathogen-specific data from the NCT07352865 adaptive trial. Recommendations are calibrated using GRADE-adapted certainty and strength, with explicit acknowledgment that the comparative evidence base remains overwhelmingly retrospective and geographically concentrated in Chinese tertiary hospitals.

Energy restriction (ER) serves as both a corrective health measure in metabolically compromised individuals and as a strategically implemented tool to enhance athletic performance and/or meet sporting weight divisions. Individuals with overweight/obesity who embark on a weight loss intervention often experience temporal improvements in metabolic health, only to be undermined by the phenomenon known as adaptive thermogenesis, and subsequent weight regain. ER in elite sport settings, whether a deliberate strategy to manipulate body composition or an unintentional failure to meet the high energy demands of intense training, can also trigger a cascade of adaptive yet energy-conserving measures across biological systems, tissues, and organs, ultimately compromising long-term health. Despite two comparatively different goals, these populations share one common requirement: a sensitive monitoring tool. Predicting problematic ER, however, remains challenging. Contemporary tools employed by practitioners and healthcare providers often suffer from a diagnostic latency, capturing 'whole-body' maladaptations only after they have passed the point of effective corrective input. In this Current Opinion, we propose that metabolomics-the comprehensive analysis of all metabolites in a biological sample-may represent a promising tool to help identify the transition from adaptation to maladaptation by capturing subtle biochemical shifts that precede whole-body changes. We explain how both populations could utilise this tool to identify when ER needs to be corrected to prevent physiological decline and protect long-term health.

The deleterious intersection of sarcopenia and age-related heart failure represents a profound global health challenge. While skeletal muscle is increasingly recognized as a major endocrine hub, cannot fully account for the persistent epigenetic changes in the aged myocardium. This points to a key mechanistic gap in the "muscle-heart" inter-organ crosstalk. Following PRISMA guidelines, this systematic review (incorporating 51 rigorous in vivo and clinical studies) maps the bimodal skeletal muscle-derived extracellular vesicle (SkM-EV) and microRNA (miRNA) axis in cardiac aging. We delineate a pathological baseline where aging and sarcopenia trigger the release of senescence-associated extracellular vesicles (EVs). These toxic vesicular payloads actively propagate myocardial inflammaging, structural remodeling, and apoptosis. Conversely, regular exercise rejuvenates by this network via an epigenetic mechanism. Mechanical loading stimulates the systemic release of "exerkines"-exercise-conditioned EVs enriched with potent cardioprotective myomiRs (e.g., miR-1, miR-133a, miR-342-5p). By systematically categorizing these findings from single-molecule downstream targets (anti-apoptosis, anti-fibrosis) to macroscopic poly-pathway synergy (antioxidant and metabolic reprogramming), we construct a comprehensive molecular roadmap of EV-mediated myocardial rejuvenation. Ultimately, deciphering this vesicular signaling network will elucidate the fundamental epigenetic mechanisms underlying "exercise as medicine," and paves the way for novel translational horizons. We propose that targeting the bimodal SkM-EV axis will accelerate the development of EV-based liquid biopsies for sarcopenic cardiomyopathy and pioneer cell-free "exercise mimetics" for frail, exercise-intolerant aging populations.

Objectives: This study aimed to investigate the effects of nature-based immersive virtual reality (IVR) applied during dry needling (DN) on pain experienced during the procedure and on immediate and short-term post-intervention outcomes, compared with traditional DN, with a primary focus on immediate post-intervention pain intensity. Methods: In this single-blinded randomized controlled trial, participants were randomly assigned to the control (DN) or experimental (DN + IVR) group. Eligible participants were healthy individuals with identifiable latent myofascial trigger points (MTPs) in the quadriceps muscles. The primary outcome was pain intensity, measured using a numerical rating scale (NRS), immediately after the intervention. Secondary outcomes included pressure pain threshold (PPT) and pain intensity at follow-up time points (1, 6, and 24 h). Linear mixed-effects models were used to analyze the effects of group, time, and their interactions. Results: A total of 40 participants (control group, n = 21, 21.7 ± 3.1 years, 15 males; experimental group, n = 19, 22.4 ± 3.7 years, 11 males) were included. The DN + IVR group showed significantly lower pain intensity immediately after the intervention than the DN group (p = 0.032). Pain intensity decreased over time in both groups (p < 0.001), with no significant intergroup differences at later time points (all p > 0.05). For PPT, a significant effect of time was observed (p < 0.001), but no significant group × time interaction was observed. Conclusions: Nature-based IVR may reduce immediate procedural pain associated with dry needling, although these effects are not sustained over time. IVR may be a useful non-pharmacological strategy for pain relief during invasive physiotherapy procedures.

Background: Myofascial temporomandibular disorders (TMDs) are associated with pain and trigger points in the masticatory muscles. Dry needling (DN) is used for myofascial pain, but evidence regarding immediate effects remains limited. Objective: This participant- and outcome-assessor-blinded randomized controlled trial evaluated immediate clinical and electromyographic effects of a single DN session targeting the masseter and anterior temporalis muscles in patients with myofascial TMD. Methods: Forty-four participants diagnosed according to the Research Diagnostic Criteria for Temporomandibular Disorders were randomly allocated to a DN group or a sham needling group. Pain intensity was assessed using a visual analog scale, and surface electromyographic activity of the masseter and anterior temporalis muscles was recorded at rest and during maximal clenching. Measurements were obtained before treatment and 10 min after the intervention. Adverse events were recorded throughout the observation period. Results: Forty participants completed the study, with 20 in each group. Pain intensity decreased significantly in the DN group compared with the sham group (p < 0.001). An uncorrected analysis indicated reduced resting activity of the left anterior temporalis muscle; however, no electromyographic variable remained significant after correction for multiple comparisons. No serious adverse events occurred. Minor transient bleeding and mild post-needling soreness were reported in a small number of participants receiving DN. Conclusions: A single DN session produced a significant immediate reduction in pain intensity in patients with myofascial TMD, without consistent immediate changes in surface electromyographic activity. Longer follow-up and repeated-session studies are needed to determine the durability and clinical relevance of these effects.

Plants host diverse microbial communities across spatially distinct foliar niches. For hemibiotrophic pathogens, the epiphytic leaf surface and the apoplast impose contrasting physiological constraints that shape infection strategies. We investigated how these environments program the transcriptional responses of a tomato pathogen, Xanthomonas perforans (Xp), and how virulence, metabolism, and regulation are coordinated across space and time during disease progression. By selectively enriching and profiling the transcriptome of a pathogen from the leaf surface and apoplast, we uncovered pronounced niche-specific and colonization stage-specific gene expression patterns. Early epiphytic colonization features activation of chemosensing and motility to promote relocation, alongside iron and phosphate acquisition, DNA and protein repair, quorum sensing, phenylalanine degradation/tyrosine conversion consistent with countering phenylpropanoid defenses, osmotic and oxidative stress mitigation, DNA exchange machinery, and type VI secretion-mediated microbial competition. Upon entry into the apoplast, Xp shifts toward active metabolism and replication, accompanied by investment in type II and III secreted virulence factor expression, together with countermeasures against host-mediated nutrient sequestration. Late apoplastic infection triggers renewed stress responses and reactivation of motility, consistent with preparation for dissemination. Across both niches, oxidative and nutrient stress responses are prominent, but the specific gene sets engaged differed by environment. Together, this work provides a mechanistic, systems-level model of how a foliar pathogen perceives and adapts to structured host niches, revealing spatiotemporal regulatory logic that coordinates survival on the leaf surface with virulence and growth in the apoplast. This framework provides broadly applicable insights into ecology and virulence programming in planta.IMPORTANCEFoliar bacterial pathogens face a series of shifting challenges as they move from the leaf surface into the protected interior. Xanthomonas perforans is a major tomato pathogen, yet how it engages in molecular dialog with the host has remained largely unclear. Reverse genetics approaches have revealed individual virulence factors, but they capture only fragments of a much larger process. By profiling pathogen transcriptomes in distinct microhabitats, we show how X. perforans reprograms its physiology during infection, adjusting motility, stress tolerance, metabolism, and virulence. This stage-resolved perspective uncovers the logic behind the infection cycle, revealing coordinated gene expression programs and regulatory modules during niche transitions. Our findings also demonstrate that ecologically similar foliar pathogens do not solve the same problems in the same way. Although X. perforans and Pseudomonas syringae occupy comparable niches, they adopt distinct metabolic and stress-response strategies, and plants perceive and respond to them differently. Such contrasts highlight how even closely related infection lifestyles can trigger unique molecular dialogs with the host. By illuminating nutrient limitations, and regulatory programs that shape infection from the outset, this work provides a systems-level view of disease establishment and identifies vulnerable points in the infection cycle that could be leveraged for targeted disease management.

Streptococcus gordonii is an alpha-hemolytic Gram-positive bacterium most commonly present in the oral cavity and gastrointestinal tract. Despite an increase in the incidence of infective endocarditis, S. gordonii cases reported on a native aortic valve are relatively uncommon. Reported cases have been linked to spondylodiskitis, intra-abdominal infection, and oral procedures. A 79-year-old male was admitted for acute kidney injury, diarrhea, and dehydration, and was found to have S. gordonii bacteremia with endocarditis. A 1.7 cm × 0.9 cm vegetation on the native aortic valve was revealed, and subsequent multiple septic emboli on brain magnetic resonance imaging (MRI) were discovered after an incident of acute mental status change. The patient was successfully treated with intravenous ceftriaxone and later required aortic valve replacement. A possible source of the bacteremia was a trigger point injection to the lumbar spine, which was consistent with MRI lumbar spine findings of a spinal abscess. However, other possible sources include a dental procedure, cholecystitis with concern for cholelithiasis, and colonoscopy with polypectomy, both incidences within eight weeks of the hospitalization. This case illustrates the importance of thorough history taking and consideration for an echocardiogram to rule out endocarditis if S. gordonii bacteremia is present, in addition to close monitoring and outpatient follow-up post-diagnosis. Considering an increasing number of procedures involving the oral cavity, spine, and abdominal area, we hope that this case will bring awareness to the diagnosis of S. gordonii endocarditis.

The rapid identification and precise quantification of cancer biomarkers are essential for the purposes of diagnosis, classification, and therapeutic intervention. Traditional molecular diagnostic methodologies, such as polymerase chain reaction (PCR), provide considerable sensitivity; however, they depend on exponential amplification and advanced instrumentation, thereby constraining their applicability for point-of-care testing. In this study, we developed a Transcription-driven CRISPR Cascade Amplification (TCCA) for one-tube detection of mRNA at the concentration of 0.6 copies/μL within just 30 min. The target-induced assembly of a three-way junction (TWJ) facilitates the generation of a split T7 promoter, which initiates transcription and activates Cas13a collateral cleavage. The activated Cas13a subsequently cleaves cascade probes and releases new trigger strands, forming a transcription-driven cascade amplification circuit. The assay enables multiplex detection of breast cancer-associated mRNA biomarkers (HBB, KRT17, and CD55) in cell lysates, demonstrating robust performance in intricate biological matrices. Furthermore, incorporation of a multiplex OR-gated logic design enables parallel target recognition, thereby enhancing detection reliability for rapid and accurate clinical diagnostics.

Allosteric effects are widespread in proteins, but predicting the impact of sequence substitutions at positions distant from ligand-binding or enzyme active sites remains challenging, as their effects are mediated through complex dynamical networks. Pinpointing such positions experimentally is labour-intensive and low throughput. Equilibrium molecular dynamics (MD) simulations are useful: analysis methods can identify functional distal sites and networks, while free energy simulations can predict effects on stability or binding, given sufficient sampling; such simulations are typically time-consuming, requiring extensive simulation of each individual mutant. Here, we introduce a dynamical-nonequilibrium MD (D-NEMD) protocol using alanine substitutions as targeted perturbations to identify distal positions that influence enzyme function. Using the well characterised class A β-lactamase KPC-2 as a test system, we show that D-NEMD distinguishes functional from non-functional sites based on whether substitutions generate structured, long-range responses that reach the active site. KPC-2 is clinically important due to its broad substrate spectrum and the emergence of resistance-associated point mutations, many of which act through non-local effects on catalytic residues. Alanine substitutions at positions 179 and 164, which disrupt the Ω-loop salt bridge, trigger persistent structural responses propagating through the protein and reach both catalytic residues and the oxyanion hole backbone. Likewise, alanine substitution at position 220 elicits pronounced responses extending to the active-site β-sheet and key loops, consistent with its known role in substrate specificity. In contrast, substitution at position 276-mutations of which have negligible kinetic impact-produces only local displacements with minimal propagation and no effect on catalytically relevant regions. These differential response patterns align with experimentally observed resistance phenotypes. D-NEMD therefore provides a fast, generalisable, and predictive approach for identifying allosterically connected distal positions. The protocol complements equilibrium MD, is straightforward to implement, and offers a tractable route for prioritising candidate sites for mechanistic study or future mutational scanning efforts.

Identification of signaling networks is an essential goal in systems biology. Here, we use CRISPR/Cas9 knockout screening (employing a whole kinome sgRNA library) to identify functionally critical protein kinases in a well-studied G α s-dependent G-protein coupled receptor (GPCR)-signaling model, namely the vasopressin V2 receptor (V2R) pathway. Screening was done using a specially-designed fluorescence-based reporter cell line with green-fluorescent protein (GFP) co-transcribed with Aqp2 , a gene whose transcription is dependent on vasopressin-mediated activation of protein kinase A (PKA). Positive regulators (n=14) included PKA-catalytic subunit α (Prkaca) and Dyrk1a ( minibrain homolog). Negative regulators (n=12) included PKA-regulatory subunit type Iα, Stk11 (catalytic subunit of liver kinase B1 [LKB1] complex), and three TGF-β receptor subunits (Tgfbr1, Tgfbr2, Tgfbr3) (see https://esbl.nhlbi.nih.gov/Databases/Kinome-CRISPR-screen/ for full list). Dyrk1a knockout cell lines failed to express AQP2 protein and exhibited a profound decrease in AQP2 mRNA. RNA-sequencing demonstrated widespread increases in cell-cycle transcripts, with a general defect in cell differentiation, accounting for AQP2 loss. TGF-β exposure to un-transformed cells results in a profound decrease in V2R and AQP2 mRNA abundance along with multiple additional transcriptional targets of V2R-PKA signaling, consistent with prior findings in TGF-β-mediated vasopressin 'escape'. Stk11/LKB1 knockout lines displayed marked increases in AQP2 protein and mRNA, even in the absence of vasopressin. RNA-sequencing showed a marked similarity between the responses to Stk11/LKB1 deletion and vasopressin exposure in untransformed cells. Phospho-proteomic data point to opposing roles of Stk11/LKB1 and PKA in the regulation of cAMP-responsive transcriptional coactivator (CRTC) proteins in the transcriptional response to V2R-PKA signaling.