Planarian flatworms represent one of the most evolutionarily informative nervous systems for an account of ancient bilaterian brains. Likewise, the unparalleled regenerative ability of planarians makes possible certain investigations of neural development, memory, and behavior that are simply impossible with other model organisms. Despite these facts, learning and memory are today underexplored in planarians, likely due in part to the controversial legacy of 20th-century experiments on the transfer of memories between individual flatworms. Here, we attempted to replicate and extend the classical conditioning experiments in planarians that were the basis of the later memory transfer work. We failed to find evidence of learning using both historical and contemporary conditioning protocols, and using computer vision methods to avoid subjectivity in manual video annotation. Our results cast doubt on the suitability of historic conditioning protocols for Girardia dorotocephala and Schmidtea mediterranea planarians. We encourage future researchers to explore more ethologically relevant conditioning paradigms to establish a behavioral foundation for investigating the molecular basis of learning.

The folding of newly synthesized proteins is assisted by an essential network of proteins known as molecular chaperones. In bacteria, the chaperones Trigger Factor (TF), DnaK/DnaJ (Hsp70 system), and GroEL/GroES (chaperonin system) are key players in proteostasis maintenance. Here, we further explore the cooperation and substrate preference within the bacterial chaperone network by performing directed evolution of GroEL/GroES in Escherichia coli lacking both TF and DnaK chaperone pathways. We found that single amino acid substitutions in GroEL were sufficient to significantly improve its chaperone activity in vivo at high temperature in the absence of TF and DnaKJ, with a combination of selected mutations further enhancing functionality. In vitro analysis showed that high-performing GroEL variants have a higher rate of ATP hydrolysis and an increased folding rate for certain substrates at low ATP concentration, thereby compensating for the significant drop in ATP level observed in vivo in cells lacking TF and DnaKJ. Analysis of substrates bound to GroEL in vivo showed that experimentally evolved chaperones likely adapted toward obligate GroEL substrates with low solubility, thus bypassing cooperation with upstream chaperones like TF and DnaKJ.

Isolation of pancreatic islets for single-cell RNA sequencing (scRNA-seq) remains technically challenging because enzymatic digestion, mechanical dissociation, and subsequent purification can cause cell loss, aggregation, and processing-induced transcriptional changes. This study aimed to adapt a practical gradient-free alternative for preparing murine pancreatic islets for downstream single-cell transcriptomic analysis while reducing the number of sample-processing steps. The workflow combined intraductal perfusion of the pancreas with 3 mL of a collagenase solution through the common bile duct, controlled enzymatic digestion at 37 °C, performed manual islet selection without density-gradient purification, and achieved gentle dissociation with Accutase and EDTA. DNase I was included in all post-digestion steps to limit cell aggregation. The protocol was evaluated using pancreatic islets from BKS.Cg-Dock7m +/+ Leprdb/J mice, a model of type 2 diabetes. Compared with Ficoll-based purification, the gradient-free protocol provided viable cell yields within a similar range while eliminating density-gradient preparation, an 18 min gradient centrifugation step, two subsequent wash centrifugations, and the associated transfer and dilution steps. The resulting scRNA-seq library showed satisfactory quality-control (QC) characteristics and retained the major endocrine populations, including β-, α-, δ-, and PP cells. The gradient-free sample also showed a lower overall stress-response score and lower expression of several immediate early genes, although heat shock- and endoplasmic reticulum (ER) stress-associated genes displayed heterogeneous patterns. These differences were considered descriptive because only one pooled scRNA-seq library was generated for each workflow. These findings support gradient-free isolation as a possible alternative to Ficoll-based purification for preparing murine pancreatic islets for scRNA-seq analysis.

Hydatid cyst-related mortality is rare and diagnostically challenging in forensic practice, particularly when cystic echinococcosis remains clinically unrecognized until death. This study aimed to characterize hydatid cyst-related deaths through integrated assessment of clinical course, macroscopic morphology, histopathology, and mechanism of death. We retrospectively reviewed 32 hydatid cyst-related deaths evaluated by the First Specialized Board of the Council of Forensic Medicine of Türkiye between 2013 and 2024. Demographic and clinical characteristics, organ involvement, macroscopic cyst features, histopathological findings, and certified mechanisms of death were assessed. Macroscopic and histopathological findings were categorized using predefined morphological classifications, and clinicopathological associations were analyzed. The median age was 27.5 years, and 71.9% were male. Hepatic involvement was present in 93.8%, while hydatid disease had not been diagnosed before the fatal event in 84.4%. Anaphylactic shock was the predominant fatal mechanism (59.4%), followed by sepsis/multiorgan failure (21.9%), hydatid-related sudden death without further mechanistic specification (9.4%), pulmonary hydatid embolism (6.3%), and direct cardiac hydatid-related sudden death (3.1%). Intact or tense cysts predominated among macroscopically evaluable cases (73.3%), whereas complete rupture was identified in 16.7%. The secondary infected histopathological phenotype was associated with sepsis-related death (OR 18.0; 95% CI 1.54-210.6; p = 0.025). Hydatid cyst-related deaths occur through distinct clinicopathological pathways, with anaphylactic shock predominating. Complete macroscopic rupture is not required for fatal hypersensitivity reactions. Integrating gross morphology, histopathology, systemic autopsy findings, and clinical context may improve postmortem mechanistic interpretation.

Peripartum cardiomyopathy (PPCM) is a rare but potentially life-threatening cause of heart failure that requires prompt recognition and exclusion of alternative etiologies. This case highlights the complementary roles of computed tomography and cardiac magnetic resonance (CMR) in the peripartum patient with acute heart failure.

Heat stress in livestock is a heeding concern, and the addition of optimum fiber to the diet is an option in balancing the metabolic heat load. This study evaluated the comparative effect of low, moderate and high dietary Neutral detergent fiber (NDF) levels on nutrient utilization, production, hormone levels, hemato-biochemical parameters, antioxidant status and stress indicator levels in mid lactation Murrah buffaloes (MY 9.2 ± 2.2, Parity 2.5 ± 1.5). Eighteen mid lactation female buffaloes were randomly allocated to three dietary treatments for 90 days: a total mixed ration (TMR) with 30.0% NDF (C1), 34.5% NDF (T1) and 37.0% NDF (T2). During the trial, THI ranged from 77.0 to 88.3, indicating that Murrah buffaloes were exposed to heat stress. Compared with the control, buffaloes fed 34.5% NDF diet exhibited a 4.5% increase in milk yield, accompanied by a 6.2% increase in NDF digestibility and a 15.4% reduction in serum NEFA concentration. A moderate dietary NDF level lowered respiratory rate, heat shock protein and increased intake, FCM, T3 and T4 levels. Plasma cortisol, albumin, globulin, BUN remained unaltered with altered fiber level except for increased total protein with high fiber. Antioxidant status remained unaffected with varying NDF supplementation except for increased SOD at moderate and high fiber levels. Milk fat and total solids increased at high NDF levels. Milk stress indicators unaffected with altered fiber levels but altered with respect to acceptable levels under stress conditions. Overall, an optimum dietary fiber level, improved nutrient utilization, production performance, antioxidant status, hormonal status and haemato-biochemical performance in lactating Murrah buffaloes.

Soil salinization, exacerbated by seawater intrusion and climate change, poses a major constraint on crop production. To cope with salt stress, plants have evolved complex mechanisms involving numerous tolerance-related genes. We identified that heat shock protein TaHSP70 acted as a positive regulator in response to salt stress in wheat. Further analysis revealed that heat shock transcription factor TaHSFA6e and NAC transcription factor TaNAC019 physically interact and function additively to activate TaHSP70 expression. Whereas a single-nucleotide T insertion within the coding sequence of TaNAC019-B reduced the transcriptional activity of TaHSP70. Overexpression of either TaHSFA6e or TaNAC019 enhanced salt tolerance in wheat. Collectively, these findings highlight the importance of the heat shock protein pathway in mediating salt stress responses in wheat.

The healing of diabetic wounds is impeded by a pathological triad of recalcitrant bacterial infections, oxidative stress, and chronic inflammation. While photothermal therapy (PTT) is a promising non-antibiotic alternative, its clinical translation is constrained by a critical efficacy-biosafety paradox: high-temperature ablation (>60 °C) induces collateral tissue damage, whereas biosafe mild-temperature PTT (<50 °C) fails to achieve complete pathogen eradication due to bacterial heat-shock defenses. To reconcile this, we engineered a multifunctional hybrid hydrogel by incorporating flower-like MoS2@ZnO heterostructures into a dynamic dextran (Dex) and poly(vinyl alcohol) (PVA) network (PD@MoS2-ZnO), executing a "sensitization-before-attack" strategy. Sustained Zn[2+] release selectively permeabilizes bacterial membranes, rendering pathogens hypersensitive to subsequent mild photothermal treatment (∼50 °C). In vitro assays demonstrated that this synergistic mechanism achieved exceptional antibacterial efficiency, eliminating 97.54% of E. coli and 98.98% of S. aureus. In a S. aureus-infected diabetic mouse model, the hydrogel treatment resulted in a 98.05% reduction in bacterial load and significantly accelerated wound closure, with the residual wound area diminishing to 1.11% after 15 days. Histological analyses further confirmed that the dressing promoted collagen deposition, re-epithelialization, and angiogenesis. This work establishes that an interfacial-sensitization platform provides a potent and biosafe strategy for managing complex diabetic wound infections without compromising host tissue viability.