Antimicrobial resistance has intensified the search for novel antimicrobial and wound-healing agents. Ludwigia octovalvis (Jacq.) P.H. Raven is traditionally used to treat infections, inflammation, and skin disorders, although its in vitro biotechnological potential remains largely unexplored. This study reports the first successful establishment of L. octovalvis callus cultures, their characterization by gas chromatography-mass spectrometry (GC-MS), and the evaluation of their antimicrobial and wound-healing activities. Friable calluses were induced from leaf and node explants of axenic seedlings using combinations of 6-benzylaminopurine (BAP), kinetin (KIN), 2,4-dichlorophenoxyacetic acid (2,4-D), and α-naphthaleneacetic acid (NAA). Optimal callus induction was achieved in leaf explants cultured on full-strength Murashige and Skoog (MS) medium supplemented with BAP (4.44 µM) and 2,4-D (0.45 µM). After six months of subculture, the calluses showed morphological uniformity and stable biomass production. Growth kinetics followed a specific growth rate of 0.047 d-1 and a doubling time of 14.54 days. GC-MS analysis identified fatty acids and phytosterols as the main components. The ethyl acetate extract significantly improved wound healing in vivo, while the methanolic extract exhibited antibacterial activity against Staphylococcus aureus and methicillin-resistant S. aureus (MRSA). These findings demonstrate that L. octovalvis callus cultures constitute a sustainable source of bioactive compounds with promising therapeutic potential.

Delayed and nonhealing fractures, affecting 5 to 10% of cases, are associated with prolonged disability and diminished quality of life. Although acute inflammation is required to initiate repair, persistent mechanical instability can sustain maladaptive immune and fibrotic responses that impede regeneration. Existing animal models do not adequately recapitulate mechanical instability, the principal driver of hypertrophic nonunion in clinical settings, thereby limiting translational relevance. We developed a murine model of delayed fracture healing using tunable intramedullary fixation to impose controlled interfragmentary strain. High-strain conditions (low-stiffness nail, 15 to 30% strain) produced enlarged calluses characterized by delayed ossification, increased fibrotic tissue (3.3-fold, P = 0.0147), and reduced biomechanical integrity (1.6-fold decrease in stiffness, P = 0.024) relative to low-strain controls (high-stiffness nail, <5% strain). Spatial transcriptomic analysis identified persistent fibrotic niches in high-strain calluses enriched with fibroblast-associated genes (e.g., Pdgfrb and Lgals3) and dysregulated macrophage-fibroblast signaling (Spp1 and Mmp9). Systemically, high-strain fractures were associated with distinct immune signatures in which CD206[+] macrophages and CD25[+] regulatory T cells predicted healing outcomes (R = 0.72, P = 0.004), suggesting early immune polarization as a determinant of repair trajectory. Elevated CD8[+] T cell responses were observed, consistent with a sustained inflammatory state associated with impaired healing. These findings identify mechanical instability as a driver of pathological immune-stromal interactions and establish a preclinical platform for investigating mechanobiology-informed therapeutic strategies. This work supports a conceptual framework in which hypertrophic nonunion is understood as a disorder arising from dysregulated interactions between mechanical cues and immune responses.

Here, we describe and characterize catastrophic pelvic fractures in Thoroughbred racehorses in California. We reviewed retrospectively racehorse autopsy reports from horses that died spontaneously or were euthanized because of complete pelvic fractures in California between 1992 and 2018. The exercise history of affected horses was compared with eventmatched control horses. Catastrophic pelvic fractures were sustained by 133 horses. Pelvic fractures were 2.7% of all musculoskeletal fatalities. Pelvic fractures occurred during training (48.0%), racing (35.0%), and non-exercise (17.0%). Horses ≤4-y-old were affected most frequently (73.5%); horses ≥5-y-old were affected less frequently (26.5%). Fractures were more common in females (68.1%) than in geldings (23.1%) and intact males (8.8%). Bilateral fracture occurred in 46.9% of horses. The ilium was fractured alone or with other pelvic bones in 90.3% of horses; 28.4% of these cases included bilateral iliac wing fractures. Preexisting periosteal calluses associated with the catastrophic fracture line were observed in 31.9% of horses with pelvic fractures. Calluses were frequently associated with ilial fractures (88.9%). Of horses with complete pelvic fracture, 25.7% died spontaneously, and 89.6% of those deaths were a consequence of massive internal bleeding from lacerated blood vessels. The horses with pelvic fractures had fewer races (spaced farther apart), exercised less intensely throughout their career, and had more lay-ups than control horses. They also had 31% less exercise time and 40% fewer events since the last lay-up than control horses. Factors associated with these events, such as preexisting injuries, should be identified to enable recommendations for diagnosis, treatment, and prevention of catastrophic pelvic fractures.

Surgical treatment for forefoot deformities in patients with rheumatoid arthritis has recently shifted toward joint-preserving procedures for all toes. In contrast, resection arthroplasty was previously more commonly performed for the lesser toes. Advances in drug therapy have improved life expectancy, resulting in an increased number of long-term survivors who had previously underwent resection arthroplasty. With increased activity levels, the recurrence of plantar callosities in the forefoot has become more frequent, often leading to gait disturbances. In general, revision surgery for such cases often involves additional bone resection; however, this approach may result in further shortening of the metatarsals and increase the risk of further recurrence. Therefore, we applied a metatarsal-shortening offset osteotomy, which we typically perform as a primary procedure, as a salvage procedure to reduce the risk of recurrence. We report three cases treated using this strategy. In all cases, the second to fifth metatarsal heads had been resected, but the resection stumps were covered with pseudo-cartilaginous tissue. This allowed a metatarsal-shortening offset osteotomy to be performed as a procedure similar to that used in primary surgery, achieving the planned correction. Consequently, both radiographic and clinical improvements were observed. These findings suggest that a metatarsal-shortening offset osteotomy may be a possible salvage option after primary resection arthroplasty for forefoot deformities in patients with rheumatoid arthritis.

The present video article demonstrates the use of Evans and Cotton osteotomies with porous titanium wedges for the treatment of pediatric flatfoot. Pediatric flatfoot is not commonly painful or disabling and usually resolves as the arch elevates spontaneously. However, in cases of flatfoot that persist despite nonoperative treatment, symptoms such as severe activity-related pain and medial foot calluses are an indication for surgical intervention3. The aims of treatment in pediatric patients are to correct anatomical deformity and reduce pain while maintaining mobility.

Soybean calluses constitute a robust in vitro model for investigating biochemical and elemental responses to external stressors. In this study, transgenic soybean calluses (Glycine max) were exposed to increasing concentrations of copper nanoparticles (CuNPs; 10, 50, and 100 µg L⁻1), and their chemical responses were investigated using an integrated online HILIC-ESI-HRMS-ICP-MS/MS platform. This hyphenated analytical approach enabled the simultaneous chromatographic separation, molecular characterization, and elemental detection of co-eluting chemical species, providing complementary metabolomic and speciation information within a single analytical workflow. The online coupling revealed marked concentration-dependent effects, with calluses exposed to 10 and 50 µg L-1 CuNPs exhibiting reduced chemical diversity compared with those exposed to 100 µg L-1. Multivariate and pathway-level analyses indicated that CuNP exposure progressively reshaped the metabolic landscape, with the most pronounced alterations observed at the highest concentration. These changes were primarily associated with cellular defense responses, redox imbalance, and membrane remodeling, as evidenced by the increased accumulation of oxylipins and lysophospholipids, including 12(13)-Ep-9-KODE, (9Z,12Z)-7,8,16-trihydroxyoctadeca-9,12-dienoic acid, and LysoPC(16:0/0:0). Importantly, the integrated speciomics strategy afforded by the online HILIC-ESI-HRMS-ICP-MS/MS platform enabled the putative annotation of metal- and heteroatom-containing species, suggesting homeostatic adjustments involving essential elements such as Mn, Mg, Zn, P, and S associated with organic ligands. These findings highlight the central role of elemental redistribution and coordination chemistry in shaping the biochemical response of soybean calluses to nanoparticle-induced stress and demonstrate the analytical potential of integrated molecular and elemental detection for investigating complex biological systems.

Since the status of C. jambhiri is "rare" and C. aurantifolia is endowed with folk medicinal properties, our study aimed at producing true-to-type seedlings for further conservation by using PGRs, ferrous sulfate, and casein hydrolysate in MS medium to induce in vitro germination and "callus" formation from seeds of C. jambhiri Lush. It also focused on evaluation of suitable species and the best type of explant for organogenesis. The present study was undertaken to develop an efficient micropropagation protocol for C. jambhiri Lush. and C. aurantifolia. The frequency of callus induction increased to its maximum when 2.0 mg/L 2,4-D and 1.0 mg/L picloram were added individually. The calli derived from 2,4-D exhibited maximum regeneration potentiality. In addition, sucrose (30 g/L), dextrose (60 g/L), and coconut water (10 mL/L and 15 mL/L) also enhanced callus induction. Regarding heavy metals, 100 ppm of Fe2SO4 exhibited maximum germination percentage (84.33%) from seeds of C. jambhiri Lush. However, maximum callus induction (50.00%) was induced from the seeds of C. aurantifolia incubated in Fe2SO4 (400 ppm). The maximum number of shoots per callus was produced (5.13) with the addition of 200 mg/L casein hydrolysate in the callus-induction medium. Pearson correlation analysis revealed a positive and significant association of the number of plantlets/calluses with shoot length and regeneration percentage, respectively. It was phenotypically observed that the tissue culture traits performed better from explants derived from C. jambhiri Lush. than C. aurantifolia when subjected to varying concentrations of PGRs, carbon sources, organic adjuvants, and heavy metals, respectively.

Fracture healing is a complex, multiscale process governed by the interaction between biological activity and mechanical environment, and fracture callus geometry represents an integrated manifestation of these processes. Although callus morphology is routinely assessed during healing, its value as an objective and predictive marker of mechanical competence and clinical outcome has not been systematically established. This review synthesizes current experimental and clinical evidence to clarify the determinants of callus geometry and to evaluate its potential as an early indicator of fracture healing. A systematic review was conducted in accordance with the PRISMA guidelines, using major databases to identify preclinical and clinical studies reporting qualitative or quantitative measures of fracture callus geometry. Eligible studies examined biological, mechanical, systemic, or pharmacologic influences on callus formation, or assessed associations between callus morphology and biomechanical strength or clinical healing outcomes. Extracted data included study design, fracture model, imaging modality, geometric parameters, and functional endpoints. Given substantial heterogeneity in models, measurement techniques, and assessment timing, findings were synthesized narratively. Forty-eight studies met the inclusion criteria. Across species and fracture models, callus geometry consistently reflected the underlying healing environment. Angiogenic and osteoanabolic interventions, including deferoxamine, platelet-rich plasma, and sclerostin inhibition, were associated with larger, more mineralized, or better-organized calluses, highlighting the importance of vascular and osteoblastic signaling. In contrast, chronic PTH or PTHrP stimulation often produced smaller yet structurally organized calluses, underscoring the need to interpret geometric measures within a biological context. Advanced imaging studies demonstrated strong associations between microarchitectural features and mechanical strength, while automated and CT-based methods improved reproducibility and translational feasibility. Non-modifiable factors such as metabolic disease, systemic inflammation, and traumatic brain injury also produced characteristic alterations in callus morphology. Overall, callus geometry emerges as a biologically meaningful and mechanically informative marker of fracture repair. Standardized, quantitative assessment of callus geometry shows promise as a candidate imaging biomarker of healing trajectory and warrants prospective clinical validation before a prognostic role can be established.

This study establishes a genotype-specific transformation system for indica rice cultivars Ranjit, Mahsuri, and Kon Joha using 35S:RUBY and CRISPR/LbCas12a constructs, enabling functional genomics studies and genetic improvement. The indica rice subspecies generally faces challenges in functional genomics and genetic improvement due to its recalcitrance to tissue culture and Agrobacterium-mediated transformation. Three indica rice varieties, Ranjit, Mahsuri, and Kon Joha, cultivated in Assam (India) were selected to optimize callus induction, regeneration, and genetic transformation. Ranjit and Mahsuri are high-yielding cultivars, whereas Kon Joha is an indigenous aromatic landrace of high commercial value. Initially, key steps, such as callus induction and regeneration, were optimized for Ranjit and Kon Joha using mature seed. Thereafter, for transformation, immature embryos were selected as explants because of their competence for agro-infection. The highest callus induction frequency of 70.33% and 90% was achieved with 3.0 and 3.5 mg L-1 of 2,4-dichlorophenoxyacetic acid (2,4-D) in Ranjit and Kon Joha, respectively. The best regeneration, in Ranjit (77%) and Kon Joha (54.5%), was achieved with 4.0 mg L-1 6-benzylaminopurine (BAP), 1.0 mg L-1 kinetin, and 0.5 mg L-1 naphthalene acetic acid (NAA). Transformation efficiencies with CRISPR/LbCas12a (OD660 = 0.8) were 5.6% (Ranjit), 7.5% (Mahsuri), and 15.33% (Kon Joha). The optimal in vitro regeneration conditions previously identified for Ranjit were adopted for Mahsuri transformation. Furthermore, a visual non-invasive reporter, RUBY, was employed to investigate the transformation of immature embryos in Ranjit and Kon Joha. The vivid red coloration in the early callusing stage indicates successful transformation events. Kon Joha plants regenerated from red calli showed a distinct phenotype with pink leaves and red roots. These findings confirm RUBY's effectiveness as a non-invasive reporter for rapid monitoring of rice transformation and underscore the importance of immature embryos in indica rice transformation.

Hydrocera triflora, a rare and endangered aquatic species of phylogenetic importance within the family Balsaminaceae, lacks any reported in vitro regeneration system. This study establishes the first complete in vitro propagation protocol for this taxon, integrating optimized sterilization, shoot induction, proliferation, and rooting. Stem explants (~ 3 cm) were disinfected using ethanol, 0.1% mercuric chloride with Tween, and 20% sodium hypochlorite, with the most effective method combining ethanol pre-wash and 0.1% mercuric chloride for 15 min, resulting in 35% contamination and 13% browning. Shoot induction reached 96% on medium supplemented with 1.0 mg L⁻¹ 6-benzylaminopurine and 0.2 mg L⁻¹ indole-3-butyric acid, producing 3-12 healthy shoots per explant with minimal callus. Moderate cytokinin concentrations optimized shoot proliferation, maximizing multiplication coefficient and vigor while avoiding excessive callusing. Rooting was highest on Murashige and Skoog medium with 0.2 mg L⁻¹ indole-3-butyric acid, where roots appeared by day 5, averaged 6.3 cm in length with 16.1 roots per plantlet, and achieved a 100% rooting rate. These results indicate that precise cytokinin-auxin balance governs organized morphogenesis in H. triflora, providing insights into the developmental plasticity and tissue-specific hormonal responses of this species. The established protocol offers a reliable platform not only for large-scale micropropagation and ex situ conservation but also for future biotechnological applications, including genetic improvement, secondary metabolite production, and translational studies in related recalcitrant aquatic species.