Search Results (157)

Anisakis simplex allergens are highly resistant to conventional processing, posing a health risk that limits the use of infested fish both for human consumption and for animal feed. To assess the safety and nutritional value of viscera silage, hake (Merluccius merluccius) viscera were processed with formic acid and stored for four months. Two experimental diets were prepared—one containing silage and another with untreated viscera—alongside a commercial control feed. Juvenile gilthead seabreams (Sparus aurata) were fed these diets for 86 days, and growth performance, muscle composition, and allergenicity were analysed. All groups achieved more than 20% weight gain, and the silage-fed fish showed the lowest residual allergenicity (15.3%), compared to the fish fed untreated viscera (24%) and commercial feed (21.9%). Silage processing caused extensive protein hydrolysis, reducing high-molecular-weight proteins, though some IgE-reactive fragments remained. Proteomic analysis detected only seabream structural and metabolic proteins, with no confirmed Anisakis allergens. These findings indicate that silage produced from Anisakis-infested viscera should not pose a higher risk to consumers than the fishmeal used in aquaculture feeds and could support the valorisation of currently discarded fish viscera. Full article

Background: The estimation of the postmortem interval (PMI) remains a complex challenge in forensic medicine. While macroscopic, biochemical, and molecular methods are well-documented, postmortem functional approaches at the organ level are largely underexplored. This pilot study investigated the feasibility of utilizing an isolated ex vivo kidney perfusion model to assess residual postmortem renal function—specifically glomerular filtration and tubular solute handling—as a potential chronological marker for PMI. Methods: Sixteen adult New Zealand rabbits were euthanized and randomly assigned to four postmortem interval groups (1, 5, 10, and 15 h). An unoxygenated, room-temperature crystalloid perfusion system was established to mimic natural postmortem decay. Initially, 32 kidneys were perfused; two were excluded due to anuria, resulting in 30 successfully analyzed kidneys. To strictly eliminate pseudoreplication bias, bilateral functional data were mathematically aggregated at the subject level, establishing the individual rabbit (n = 16) as the statistical unit. Results: Following statistical adjustment at the subject level, none of the measured functional parameters exhibited statistically significant chronological variation across the postmortem intervals (all p > 0.05; statistical significance defined as p < 0.05). Glomerular filtration was profoundly depressed across all groups, with adjusted inulin clearance ranging between 0.0031 and 0.0086 mL/min/g (peaking nonsignificantly at 10 h). Furthermore, active tubular reabsorption was virtually nonexistent; calculated reabsorbed loads for evaluated solutes, particularly potassium and sodium, yielded predominantly negative values. This phenomenon indicates a complete absence of physiological active reabsorption, reflecting instead a massive passive leakage of intracellular electrolytes into the tubular fluid due to cellular autolysis. Conclusions: Within this specific experimental setup, the isolated kidney perfusion model failed to demonstrate reproducible, time-dependent renal function useful for PMI estimation. These findings indirectly suggest that, unlike the prolonged supravital physiological resilience observed in skeletal muscle, highly metabolically active renal tissue rapidly loses its complex functional capacity following somatic death. Future studies exploring supravital renal function should consider targeting the immediate early postmortem period (0–1 h) or integrating advanced organ preservation techniques to unmask residual cellular capabilities. Full article

The use of postmortem (autopsy) material in fundamental and applied biomedical research significantly facilitates the collection of biomaterial for statistically robust sample cohorts. However, natural adaptive processes to developing cellular stress in the early postmortem period, caused by oxygen and nutrient deprivation, trigger the activation of numerous genes promoting cell survival under stress. Many of these activated pathways are also crucial for tumor cell survival in vivo, as evidenced by various transcriptomic studies. This study aimed to investigate the potential influence of postmortem interval (PMI) duration on gene expression in normal and tumor tissues. Using a model of chemically induced hepatocellular carcinoma in mouse liver, we comparatively analyzed the dynamics of transcript levels for several genes (BRCA1, BRCA2, CHEK1, CHEK2, ATM, CDK12) in paired samples of normal and tumor tissue over a 24-h PMI using RT-qPCR. In normal tissue, gene expression increased significantly, while tumor tissue demonstrated relative transcriptional stability, with no substantial changes in the studied transcript levels. A critical finding was the observed convergence of expression profiles: initial differences between the tissues were completely eliminated by 24 h PMI. This pattern developed despite formally adequate RNA quality (RQN) and the absence of clear signs of progressive autolysis in histology, indicating the insufficiency of standard quality criteria for detecting postmortem changes. These findings collectively underscore the critical importance of minimizing and controlling PMI during the biobanking of oncological samples for reliable transcriptomic research. Full article

During alcoholic fermentations, some non-Saccharomyces yeasts are often displaced by Saccharomyces cerevisiae. It remains unclear whether this displacement is mediated by metabolites produced by S. cerevisiae or depends on cell–cell contact. This study evaluated the effects of extracellular metabolites produced by S. cerevisiae on the growth and fermentative performance of Kluyveromyces marxianus isolated from mezcal fermentations. The development of both yeasts was evaluated in monocultures and in co-cultures with physical contact. Indirect interaction was also tested by exchanging cell-free fermented media. The growth and fermentative response of K. marxianus in cell-free S. cerevisiae-fermented medium showed modulation that depended on the growth phase during which the exchange was performed. The exchange performed at 6 h (exponential phase) limited the maximum growth of K. marxianus and resulted in lower fermentative performance. When the exchange was done during the stationary phase (17.5 h), K. marxianus exhibited a longer stationary phase and better fermentative performance. Finally, when the exchange was performed at 24 h (the beginning of the death phase), the effects on survival and fermentative performance were less pronounced. Furthermore, co-culture with cell–cell contact showed that direct competition and/or mechanisms dependent on physical contact intensify the displacement of K. marxianus. The results show that direct cell–cell contact promotes greater inhibition of K. marxianus by S. cerevisiae, which is relevant for the design of mixed fermentations aimed at achieving a greater contribution of non-Saccharomyces yeasts to the organoleptic characteristics of alcoholic beverages. Full article

In this experiment, we used a late 17th century embalming protocol for the long-term preservation (7.2 years) of an infant-sized piglet in order to evaluate the success of this technique over a period of several years. According to the description of the French anatomist Penicher (published in 1699), an 8.8 kg female piglet corpse was treated with a broad spectrum of herbs, seeds, leaves, flowers and dried berries, along with an alcohol-based fluid following subtotal exenteration and a reduction in peripheral muscle mass. The further process of this dry embalming technique was monitored by visual, tactile and olfactory evaluation of the embalmed body, along with a record of the body weight. Repeatedly taking samples from the skin and soft tissues provided insight into eventual changes on a histomorphological level and two whole-body CT scans complemented the evaluation of the internal changes within the corpse, which was eventually examined at autopsy. On the macroscopic level, we recorded slight signs of autolysis and very mild putrefaction within the first few weeks and a very well preserved and stable body over the subsequent years of evaluation. In parallel, we noted a gradual loss of fluid, as shown by a reduction in the body weight. This occurred faster in the first year (reduction in body weight by ca. 25%) than in the following ca. 4 years (with another ca. 25% loss of body weight). The CT scans showed stable osseous and soft tissue structures, while the few remaining internal organs that had been left inside the body after initial evisceration (such as kidneys and internal genitalia) had already completely disappeared after approx. 1 year. On the histological level, the histoanatomy of skin, subcutis and muscle remained intact over the entire observation period. A loss of epidermal cell nuclei was not noted before day 1772 and there were only slight signs of adipocire formation of fat tissue at the end point of observation (day 2634). In summary, we can confirm that excellent body preservation of external skin and soft tissue was maintained over a considerably long period (in this case, 7.2 years) using the applied protocol of dry embalming, but a complete loss of residual internal organs/structures beyond skin, subcutaneous fat tissue and muscle. Previous observations of the excellent preservation of an infant mummy that underwent the dry embalming procedure are very plausible. Full article

Postharvest autolysis severely compromises the commercial value of fresh Dictyophora rubrovolvata. This study integrated physiological, ultrastructural, and metabolomic analyses to elucidate the underlying mechanism. Results indicated a continuous decline in cellular adenosine triphosphate levels during storage, leading to an energy crisis and triggering cellular stress responses. Metabolomic analysis revealed that the fruiting bodies activate pathways such as glycolysis and the pentose phosphate pathway through metabolic reprogramming to maintain homeostasis. However, the intensifying energy crisis inhibited Calcium ion ATPase activity, disrupting ion homeostasis and leading to Ca²⁺ influx. This activated phospholipases and initiated membrane lipid degradation, accompanied by a burst of reactive oxygen species and elevated levels of H₂O₂ and malondialdehyde, creating a vicious cycle of oxidative stress. Concurrently, cell wall components (chitin, β-1,3-glucan, cellulose) are accelerated in degradation due to the upregulation of corresponding hydrolases. Transmission electron microscopy confirmed progressive disintegration of cellular structures, including mitochondria, the plasma membrane, and the cell wall. These findings establish an “energy-membrane lipid-cell wall” cascade framework, revealing that D. rubrovolvata autolysis is an active, orderly form of programmed cell death under energy stress. This study provides new insights into the physiological mechanisms of postharvest quality deterioration in edible fungi. Full article

Cheese ripening is slow and costly, driving interest in accelerating maturation. This study aimed to isolate a safe, efficient adjunct starter from traditional Sichuan yak yoghurt, a niche rich in stress-adapted lactic acid bacteria. From 295 isolates, 15 strains tolerant to high salt, low pH, and low temperature were selected. Using acidification, autolysis, proteolysis, and peptidase activity as indices, principal component analysis identified Limosilactobacillus fermentum 270 as the best candidate. Phenotypic assays showed no haemolysis, gelatin liquefaction, indole production, or amino acid decarboxylase activity. Whole-genome sequencing confirmed species identity and revealed 52 protease/peptidase genes, complete pathways for diacetyl/acetoin biosynthesis and branched-chain amino acid conversion, and no functional biogenic amine synthesis genes. Stress-related genes (F-ATPase, glycine-betaine transport, cold-shock proteins) support cheese adaptability. Antibiotic resistance gene homologs were mainly chromosomal and unlinked to mobile genetic elements; a functional CRISPR-Cas system lowers horizontal transfer risk. The strain was developed as a freeze-dried direct-vat starter (97.3% viability). Orthogonal optimisation of yak Gouda cheese-making defined best conditions: 0.018% adjunct, 45 min acidification, pH 5.8, and 30% curd washing. L. fermentum 270 thus combines proteolytic, flavour-enhancing, genetic safety, and processing traits, offering a promising adjunct for accelerated cheese ripening. Full article

The multiple-transferable resistance protein (MtrR) is a transcriptional repressor of the mtrCDE-encoded drug efflux pump and Type IV pilus biosynthesis (pilM), and an activator of penicillin-binding protein 1 (ponA) expression in Neisseria gonorrhoeae. Previously published microarray data suggested that MtrR is also an activator of ltgA expression in the gonococcus. LtgA is a lytic transglycosylase responsible for approximately half of recycled peptidoglycan fragments and released peptidoglycan-derived cytotoxins, which cause ciliary damage and induce specific inflammatory responses. The fragments generated by LtgA during peptidoglycan remodeling can either be recognized by the permease AmpG for uptake into the bacterial cytoplasm and recycled for new cell wall growth and general metabolism or released into the external milieu. Therefore, we sought to define the capacity of MtrR to regulate LtgA expression in gonococci. We show that MtrR binds to the ltgA promoter region in a concentration-dependent manner, and that this binding results both in increased ltgA mRNA transcription and LtgA protein levels during exponential growth. Deletion of mtrR in N. gonorrhoeae decreased peptidoglycan monomer release from growing cells and increased autolysis. These results suggest that MtrR regulation of ltgA impacts peptidoglycan-derived cytotoxin release and autolysis in the gonococcus. This study suggests a central role of MtrR in coordinating aspects of the cellular envelope that may contribute to gonococcal pathogenesis. Full article

Background: Decompressive craniectomy (DC) is a life-saving intervention for refractory intracranial pressure (ICP). While outcomes in adults are well documented, pediatric data, especially concerning pupillomotor dysfunction, remain limited. Anisocoria is generally considered a marker of severe neurological compromise, but its clinical relevance in children undergoing DC has not been adequately studied. Methods: We retrospectively reviewed 25 pediatric patients treated with DC between 2004 and 2024. Demographic, radiological and clinical data included age, sex, hospital stay, operative time, etiology, side of craniectomy, preoperative midline (ML) shift, Marshall score, Rotterdam score, Glasgow Coma Scale (GCS) and pupillary status before surgery. Functional outcomes were assessed using the pediatric version of the Glasgow Outcome Scale Extended (pGOS-E) at discharge, after 3 months, 1, 2 and 4 years. Results: The majority of patients were school-aged children with a median age of 10 (range 0–17) years. Traumatic brain injury accounted for 16 cases and represented the leading etiology for DC. Pupillomotor dysfunction (anisocoria or bilateral fixed pupillary dilatation) was observed in 15 of 25 patients, 47% of whom died during hospitalization, demonstrating a significant association with in-hospital mortality (p = 0.02). However, survivors with primary pupillomotor dysfunction demonstrated a favorable recovery at 12 months with a median pGOS-E of 6 (range 4–8), indicating moderate disability. A preoperative ML-shift > 5 mm was not associated with lower pGOS-E scores during follow-up (p > 0.05). Bone flap autolysis was observed in 12 out of 14 children (86%) receiving autologous cranioplasty, and 8 (57%) patients required revision surgery with synthetic material. Conclusions: In pediatric patients, pupillomotor dysfunction is associated with higher early mortality but does not reliably exclude favorable long-term outcomes. Compared with adult cohorts, children appear to have a greater potential for neurological recovery, suggesting that severe initial clinical findings alone should not preclude timely surgical intervention. Full article

Half-smooth tongue sole has high nutritional value due to its delicious meat and high protein content. However, its high protein content makes it highly susceptible to spoilage caused by microbial action. This study utilized plasma-activated water to pretreat half-smooth tongue sole, which was then subjected to various packaging methods: CK (air packaging), VP (vacuum packaging), MAP1 (75% CO₂/5% O₂/20% N₂), MAP2 (20% CO₂/5% O₂/75% N₂), and MAP3 (75% CO₂/10% O₂/15% N₂). The packaged samples were stored at −1 °C. Preservation efficacy was assessed by monitoring changes in microbial counts and physicochemical quality indicators throughout storage. The findings revealed a progressive increase in microbial counts, a deterioration in fish quality, and a darkening of color over extended storage periods. During superchilling storage, the increase in total volatile basic nitrogen (TVB-N) and K value was markedly reduced in the MAP1 group. Regarding protein stability, the MAP1 group exhibited a slower rise in carbonyl content as well as a slower reduction in total sulfhydryl content, further confirming its superior preservation effect. Moreover, this group demonstrated excellence in maintaining the secondary and tertiary structures of myofibrillar proteins, thereby minimizing the structural damage of fish during superchilling storage. In summary, based on observed microbial and protein changes, MAP1 (75% CO₂/5% O₂/20% N₂) was the most effective in preserving quality and extending shelf life. Full article

Background. Determining the cause and manner of death in scenes involving multiple and putrified bodies found in the same environment is a real challenge for forensic pathologists. While common scenarios include fires, vehicle crashes, and natural disasters, one of the most common causes is drug intoxication or poisoning, and the scene must be carefully evaluated based on circumstantial evidence. Carbon monoxide (CO) (also called “the silent killer”) remains one of the leading agents capable of producing simultaneous fatalities. In multi-body scenes, distinguishing between homicide–suicide, double suicide, and accidental deaths adds further complexity. The aim of this study is to highlight the limitations of toxicological and pathological investigations in advanced putrefaction and to emphasize the role of scene investigation in the interpretation of suspected CO-related deaths. Methods. The authors report a case of suspected CO intoxication involving two bodies in an advanced stage of decomposition recovered from the same room. The scene investigation, coupled with the presence of a malfunctioning combustion source, raised suspicion of CO exposure; however, analytical interpretation was severely constrained by the altered condition of biological samples. Results. Advanced decomposition magnifies these challenges. Putrefactive changes can mimic traumatic injuries, hide hypostasis, and compromise both macroscopic and microscopic evaluations due to autolysis and gas formation. Toxicological investigations are frequently hindered by the degradation or absence of key biological matrices such as blood, cavity fluids, or vitreous humor, rendering carboxyhaemoglobin quantification unreliable or impossible. These limitations may lead to incorrect medico-legal conclusions. Conclusions. Determining the cause and manner of death in complex multi-body scenes requires careful evaluation of circumstantial evidence and scene investigation, particularly when advanced decomposition compromises biological analyses and toxicological interpretation. Full article

Wine lees, a byproduct of winemaking, represent an underutilized source of β-glucans with potential functional applications in food. This study aimed to extract β-glucans using two methods—acid–alkaline treatment and autolysis assisted by ultrasound—and evaluate their effects when incorporated into low-fat stirred yogurt. The extracted β-glucans were added at a concentration of 0.3% (w/w), and rheological measurements were conducted over 20 days of storage. All yogurt samples showed shear-thinning behavior, with apparent viscosity decreasing from 10⁵ mPa·s at low shear rates (0.1 s⁻¹) to 10³ mPa·s at 100 s⁻¹. Yogurt with β-glucans from autolysis retained higher viscosity and viscoelastic moduli (G′ and G″), indicating better structural integrity. Time-dependent tests showed up to 45% decrease in shear stress over 10 min of continuous shearing in the sample with chemically extracted β-glucans, compared to only 28% for autolysis-derived ones. Oscillatory tests confirmed that all samples behaved as weak gels (G′ > G″). These findings suggest that β-glucans obtained via autolysis can improve the textural stability of yogurt, offering potential for functional dairy development and valorization of wine industry byproducts. Full article

Quantifying tissue, molecular, and structural integrity is essential for biobank development. However, current assessment methods either involve destructive testing that depletes valuable biospecimens or rely on manual evaluations, which are not scalable and lead to interindividual variation. To overcome these challenges, we present PathQC, a deep-learning framework that directly predicts the tissue RNA Integrity Number (RIN) and the extent of autolysis from hematoxylin and eosin (H & E)-stained whole-slide images of normal tissue biopsies. Advancing over prior QC methods focused on imaging quality control, PathQC provides sample-quality control through the direct quantification of molecular integrity (RIN) and structural degradation (autolysis). PathQC first extracts morphological features from the slide using a recently developed digital pathology foundation model (UNI), followed by a supervised model that learns to predict RNA Integrity Number and autolysis scores from these morphological features. PathQC is trained on and applied to the Genotype-Tissue Expression (GTEx) cohort, which comprises 25,306 non-diseased post-mortem samples across 29 tissues from 970 donors, when paired ground-truth RIN and autolysis scores were available. Here, PathQC predicted RIN with an average Pearson correlation of 0.47 and an autolysis score of 0.45, with notably high performance using adrenal gland tissue (R = 0.82) for RIN and colon tissue (R = 0.83) for autolysis. We provide a pan-tissue model for predicting RIN and autolysis scores for new slides from any tissue type (GitHub). Overall, PathQC enables a scalable assessment of tissue molecular and structural integrity from routine H & E images, enhancing biobank quality control and retrospective analyses across 29 tissues and multiple collection sites. Full article

Bacterial biofilms are critical contributors to chronic infections and antimicrobial resistance. Among the diverse extracellular matrix components, extracellular DNA (eDNA) and amyloid proteins have recently emerged as pivotal structural and functional molecules. Both individually contribute to biofilm stability and antibiotic tolerance, yet their cooperative roles remain underappreciated. This review aims to summarize current knowledge on the origins and functions of eDNA and amyloid proteins in biofilms, to highlight their molecular interactions, and to discuss how their synergistic effects promote biofilm-mediated resistance to antimicrobial agents. A comprehensive literature search was conducted using PubMed, Scopus, and Web of Science databases up to September 2025. Keywords included “biofilm”, “extracellular DNA”, “amyloid proteins”, “matrix”, and “antimicrobial resistance”. Relevant original research and review articles were systematically screened and critically analyzed to integrate emerging evidence on eDNA–amyloid interactions in bacterial biofilms. Current studies demonstrate that eDNA originates primarily from autolysis, active secretion, and host-derived DNA, while amyloid proteins are produced by multiple bacterial species, including Escherichia coli (curli), Pseudomonas aeruginosa (Fap), Bacillus subtilis (TasA), and Staphylococcus aureus (phenol-soluble modulins). Both molecules independently strengthen biofilm integrity and provide protective functions against antimicrobial agents. Importantly, recent evidence shows that eDNA can act as a nucleation template for amyloid fibrillation, while amyloid fibers stabilize and protect eDNA from degradation, creating a dense extracellular network. This synergistic eDNA–amyloid assembly enhances biofilm robustness, impedes antibiotic penetration, sequesters antimicrobial peptides, protects persister cells, and facilitates horizontal gene transfer of resistance determinants. The interplay between eDNA and amyloid proteins represents a central but underexplored mechanism driving biofilm-mediated antimicrobial resistance. Understanding this cooperative network not only deepens our mechanistic insights into bacterial pathogenesis but also highlights novel therapeutic targets. Strategies that disrupt eDNA–amyloid interactions may offer promising avenues for combating persistent biofilm-associated infections. Full article

Viral infections in humans and animals are increasing, and retrospective studies using formalin-fixed, paraffin-embedded (FFPE) samples reveal recurring outbreaks over past decades. However, the impact of pre-analytical factors like fixation and autolysis on immunohistochemistry (IHC) remains insufficiently understood. To examine how autolysis, fixation duration (6–72 h) and formalin concentration (2.5–25%) influence histology and IHC of canine distemper virus (CDV, Morbillivirus canis), interferon-β (IFN-β), and selected IFN-stimulated genes (ISGs), the study was conducted using an in vitro model based on persistently CDV-infected and non-infected DH82 cells (canine histiocytic sarcoma cell line). Autolysis led to a progressive loss of cell morphology, whereas formalin fixation had minimal impact. CDV nucleoprotein, ISG15, and myxovirus resistance protein (Mx) showed stable immunohistochemical signals across all fixation conditions and remained detectable after prolonged autolysis. CDV infection upregulated ISG15 and Mx. In contrast, IFN-β and phosphorylated protein kinase R (pPKR) exhibited variable staining and did not distinguish infected from non-infected samples. Overall, autolysis had a stronger negative impact on IHC signal quality than fixation parameters. Despite the limitations of the in vitro model, the robustness of CDV, ISG15, and Mx under suboptimal conditions highlights their potential utility as virus-sensing markers in FFPE material. Full article