Search Results (46)

The enzymatic coagulation of milk, crucial in cheese production, entails the hydrolysis of κ-casein and subsequent micelle aggregation. Conventional assessment standards, such as the Berridge method, depend on visual inspection and are susceptible to operator bias. Recent methods for the identification of milk-clotting time rely on optical, ultrasonic, and image-based technologies. In the present work, the composition of milk was evaluated through standard methods from ISO and AOAC. Milk coagulation time (MCT) was measured through viscosimetry, Berridge’s operator-driven technique, and a machine learning approach employing computer vision. Coagulation was additionally observed using the Optigraph, which measures micellar aggregation through near-infrared light attenuation for immediate analysis. Sheep milk samples were analysed for their composition and coagulation characteristics. Coagulation times, assessed via Berridge (BOB), demonstrated high correlation (R² = 0.9888) with viscosimetry (Visc) and machine learning (ML). Increased levels of protein and casein were linked to extended MCT, whereas lower pH levels sped up coagulation. The calcium content did not have a notable impact. Optigraph assessments validated variations in firmness and aggregation rate. Principal Component Analysis (PCA) identified significant correlations between total solids, casein, and MCT techniques. Estimates from ML-based MCT closely align with those from operator-based methods, confirming its dependability. This research emphasises ML as a powerful, automated method for evaluating milk coagulation, presenting a compelling substitute for conventional approaches. Full article

Acute ischemic stroke (AIS) is one of the leading global causes of mortality and morbidity. Clearer understanding of stroke etiology is a major clinical objective to determine appropriate strategies for secondary stroke prevention. Histological and molecular analysis of clots retrieved during mechanical thrombectomy (MT) in AIS offers a unique opportunity to study clot composition and its relation to stroke etiology. The field of clot composition analysis has undergone substantial growth in recent years, driven in part by the establishment of MT as the standard of care, as well as its expanding indications. Although many features differ between large-artery atherosclerosis (LAA) and cardioembolic (CE) clots, application of these findings to predicting stroke etiology at a clinical level remains challenging. Moreover, a significant number of patients have multiple comorbidities or suffer a cryptogenic subtype. Next-generation techniques such as multiomic sequencing offer a powerful potential to elevate our understanding of clot pathology and provide the level of granularity required for clinical diagnosis and management. Herein, we provide an updated review of the current state of the field by exploring stroke etiologies and their relationship to clot pathology, including classic histologic features as well as more recent, emerging results from proteomic and transcriptomic analyses. Full article

The main objective of this research consists in finding a rapid method for cheese lipidomics based on NMR data. This study plays an important role in differentiation and characterization of cheese samples in accordance with fat composition, especially in the case of fat substitution with exogenous animal or vegetal fat. Our findings play an important role in relation to religious requirements regarding non-allowed foods (pork fat, for example, in some cultures) and in the correct characterization of foods according to their lipidic profile. The approach consists in establishing a fingerprint region (0.86–0.93 ppm from ¹H-NMR spectra) and then creating a database of the results obtained. The evaluation of the long-chain saturated fatty acids and the saturated short-chain fatty acids (C4 to C8) was established with a newly developed set of equations that make the computation possible even when mixtures of fats from different sources are present. This was accomplished by developing a new method for quantification of the fatty acid composition of different types of cheese, based on ¹H-NMR spectroscopy. Principal component analysis (PCA) was applied to 40 cheese samples with varying degrees (0%, 5%, 12%, or 15%) of milk fat substitution (pork fat, vegetable fat, hydrogenated oils) and different clotting agents (calcium chloride or citric acid). The best sample discrimination was achieved using fatty acid profiles estimated from ¹H-NMR data (using a total of six variables), explaining 89.7% of the total variance. Clear separation was observed between samples containing only milk fat and those with added fats. These results demonstrate that the integration of ¹H-NMR spectroscopy with principal component analysis (PCA) provides a reliable approach for discriminating cheese samples according to their fat composition. Full article

Consumer interest in vegetarian, ethical, and clean-label foods is reviving the use of plant-derived milk coagulants. Cardosins from Cynara cardunculus (“thistle”) are aspartic proteases with strong clotting activity, yet their technological impact in cheese remains under-explored. This study compared a commercial thistle extract (PC) with traditional bovine rennet rich in chymosin (AC) during manufacture and 60-day ripening of Caciotta cheese. Classical compositional assays (ripening index, texture profile, color, solubility) were integrated with scanning electron microscopy, three-dimensional surface reconstruction, and descriptive sensory analysis. AC cheeses displayed slower but sustained proteolysis, yielding a higher and more linear ripening index, softer body, greater solubility, and brighter, more yellow appearance. Imaging revealed a continuous protein matrix with uniformly distributed, larger pores, consistent with a dairy-like sensory profile dominated by milky and umami notes. Conversely, PC cheeses underwent rapid early proteolysis that plateaued, producing firmer, chewier curds with lower solubility and darker color. Micrographs showed a fragmented matrix with smaller, heterogeneous pores; sensory evaluation highlighted vegetal, bitter, and astringent attributes. The data demonstrate that thistle coagulant can successfully replace animal rennet but generates cheeses with distinct structural and sensory fingerprints. The optimization of process parameters is therefore required when targeting specific product styles. Full article

(1) Background: This study evaluated the potential of an aqueous extract from Quercus robur L. leaves for chronic wound healing. Its composition, rich in bioactive compounds (tannins and flavonoids), confers antioxidant and antimicrobial properties. (2) Methods: The toxicity and ability of the extract to enhance cell migration were tested in human keratinocytes (HaCaT cell line). Additionally, a proteomic analysis was performed on treated cells. (3) Results: The extract exhibited low cytotoxicity (IC50 = 943 µg·mL⁻¹) compared to other plant extracts. At 5 mg·mL⁻¹, it significantly accelerated wound closure at 8 h, surpassing negative control and Reoxcare; however, results were comparable at 12 h. Proteomic analysis identified 117 differentially expressed proteins (21 upregulated, 96 downregulated) involved in essential processes such as cell migration, blood clotting, and cholesterol biosynthesis. Specifically, the extract increased the expression of CYP51A1, LSS, and SQLE, while inhibiting Delta (14)-sterol reductase, key enzymes in cholesterol metabolism, suggesting a potential mechanism for tissue regeneration. (4) Conclusions: The aqueous extract of Q. robur leaves shows promise as a natural therapeutic agent for chronic wound healing, potentially aiding tissue regeneration and modulation of cholesterol metabolism. Full article

Different treatments have been proposed for morphofunctional bone repair; however, they are not always efficient and have limitations. In this experimental study, we present matrix composites with a possible synergistic effect acting with scaffolds for bone growth and use of photobiomodulation (PBM) to accelerate this tissue repair. Thus, the objective was to evaluate the effect of PBM in the repair of a long bone (tibia) of rats filled with biomimetic collagen matrices with nanohydroxyapatite and heterologous fibrin biopolymer (FB). Forty-eight rats were distributed into eight groups (n = six each): Blood Clot (BC), Blood Clot + PBM (BCP), Matrix (M), Matrix + PBM (MP), Fibrin Biopolymer (FB), Fibrin Biopolymer + PBM (FBP), Matrix + FB (MFB), Matrix + FB + PBM (MFBP). A 2.0 mm bone defect was created in the proximal third of the left tibia. The BCP, MP, FBP, and MFBP groups underwent PBM during surgery and maintained twice a week until euthanasia at 42 days. Microcomputed tomography (micro-CT), histomorphological and histomorphometric analyses were performed. Micro-CT analysis revealed that PBM influenced cortical interposition between the remnant and newly formed bone. Histologically, no exacerbated inflammatory focus or foreign body-type granulomatous reaction was observed in any group; however, a vast collagenous matrix with a more oriented and thicker spatial conformation was observed in the PBM-treated groups. Histomorphometrically, the BCP, MP, and MFBP groups showed significantly higher values compared to the other groups. Specifically, the BC group presented a mean bone tissue density of 68.33 ± 7.394, while the BCP and MP groups showed 99.83 ± 11.87 and 99.67 ± 20.58, respectively (p < 0.05). Qualitative analysis of collagen fibers indicated enhanced organization and maturation in PBM-treated groups. This study concluded that the association of PBM in the repair of long bones in rats, filled with biomimetic collagen matrices with nanohydroxyapatite and fibrin biopolymer, presented results that contribute to the improvement of bone growth, together with the association of scaffolds. Full article

Background: Variability in recanalization success during endovascular treatment for acute ischemic stroke (AIS) has led to increased interests in thrombus composition and associated cellular materials. While evidence suggests that bacteria may influence thrombus characteristics, limited data exist on microbiological profiles of thrombi in stroke patients. Objectives: Characterization of bacterial communities present in thrombi of AIS patients undergoing mechanical thrombectomy, providing insights into microbial contributions to stroke pathogenesis and treatment outcomes. Methods: Thrombi were collected from 20 AIS patients. After extracting metagenome, 16S rDNA sequencing was performed. Bioinformatic analysis included taxonomy and diversity assessments. The presence of bacterial DNA and viable bacteria in thrombi was validated using polymerase chain reaction (PCR) and bacterial culturing followed by matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) analysis, respectively. Results: 16S rDNA was amplified in 19/20 thrombi (95%). Analysis identified a diverse microbial community, with Corynebacterium spp. as the most prevalent genus, followed by Staphylococcus spp., Bifidobacterium spp., Methylobacterium spp., and Anaerococcus spp. Alpha diversity analyses (Shannon index: 4.0–6.0 and Simpson index: 0.8–1.0) revealed moderate to high microbial diversity across samples; beta diversity demonstrated distinct clustering, indicating inter-patient variability in microbial profiles. PCR confirmed the presence of DNA specific to dominant bacterial taxa identified through sequencing. Culturing showed the presence of Staphylococcus epidermidis and Enterococcus faecalis in some clots as identified through MALDI analysis. Conclusions: This study shows bacterial communities present in AIS patients’ thrombi, suggesting a potential link between microbial signatures and thrombus characteristics. Full article

Milk coagulants play a crucial role in defining curd characteristics. The objective of this study was to compare the coagulation dynamics of two commonly used coagulants in cheesemaking: cardoon flower extract (Cynara cardunculus L.) and commercial chymosin, using sheep milk from four different origins in the Baixo Alentejo region of Portugal, as the substrate. Milk composition was determined using the MilkoScan 133B, while the milk-clotting time (MCT) was measured following ISO 23058/IDF 199:2006 guidelines with slight modifications and coagulation kinetics, and technological properties were evaluated using the Optigraph apparatus. The results demonstrate that the type of coagulant impacts the coagulation properties of sheep milk. Pearson’s correlation analysis indicated that milk samples with higher protein content exhibited longer coagulation times but resulted in firmer curds. On the other hand, the use of cardoon flower extract introduced greater variability compared to chymosin, with a delayed onset of coagulation, reduced curd firmness, and increased variability in enzymatic kinetics. These results suggest that cardoon extract, while traditional, introduces greater heterogeneity in curd formation compared to the more consistent action of chymosin. Full article

The spiny mouse (Acomys species) is capable of scarless wound regeneration through largely yet unknown mechanisms. To investigate whether this capacity is related to peculiarities of the hemostatic system, we studied the blood of Acomys cahirinus in comparison to Mus musculus (Balb/c) to reveal differences in blood composition and clotting in both males and females. In response to surgical manipulations, blood clots formed in wounds of Acomys comprised a stronger hemostatic seal with reduced surgical bleeding in comparison with Balb/c. Acomys demonstrated notably shorter tail bleeding times and elevated clottable fibrinogen levels. Histological analysis revealed that clots from Acomys blood had densely packed fibrin-rich clots with pronounced fibrin segregation from erythrocytes. Acomys exhibited superior plasma clot stiffness as revealed with thromboelastography. The latter two characteristics are likely due to hyperfibrinogenemia. Light transmission platelet aggregometry demonstrated that ADP-induced platelet aggregates in Acomys males are stable, unlike the aggregates formed in the plasma of Balb/c undergoing progressive disaggregation over time. There were no apparent distinctions in platelet contractility and baseline expression of phosphatidylserine. Hematological profiling revealed a reduced erythrocytes count but increased mean corpuscular volume and hemoglobin content in Acomys. These results demonstrate the distinctive hemostatic potential of Acomys cahirinus, which may contribute to their remarkable regenerative capacity. Full article

Objectives: To evaluate the prognostic utility of CT-imaging-derived biomarkers in distinguishing acute pulmonary embolism (PE) resolution and its progression to chronic PE, as well as their association with clot burden. Materials and Methods: We utilized a cohort of 45 patients (19 male (42.2%)) and 96 corresponding CT scans with exertional dyspnea following an acute PE. These patients were referred for invasive cardiopulmonary exercise testing (CPET) at the University of Pittsburgh Medical Center from 2018 to 2022, for whom we have ground truth classification of chronic PE, as well as CT-derived features related to body composition, cardiopulmonary vasculature, and PE clot burden using artificial intelligence (AI) algorithms. We applied Lasso regularization to select parameters, followed by (1) Ordinary Least Squares (OLS) regressions to analyze the relationship between clot burden and the selected parameters and (2) logistic regressions to differentiate between chronic and resolved patients. Results: Several body composition and cardiopulmonary factors showed statistically significant association with clot burden. A multivariate model based on cardiopulmonary features demonstrated superior performance in predicting PE resolution (AUC: 0.83, 95% CI: 0.71–0.95), indicating significant associations between airway ratio (negative correlation), aorta diameter, and heart volume (positive correlation) with PE resolution. Other multivariate models integrating demographic features showed comparable performance, while models solely based on body composition and baseline clot burden demonstrated inferior performance. Conclusions: Our analysis suggests that cardiopulmonary and demographic features hold prognostic value for predicting PE resolution, whereas body composition and baseline clot burden do not. Clinical Relevance: Our identified prognostic factors may facilitate the follow-up procedures for patients diagnosed with acute PE. Full article

Portal vein thrombosis (PVT) represents a restriction or occlusion of the portal vein by a blood clot, which can appear in liver cirrhosis, inherited or acquired thrombophilia, malignancies, abdominal infection, abdominal inflammation, and injury to the portal vein; it can evolve to local venous extension, recanalization, or portal cavernoma (PC). This research represents an observational study of patients admitted with a diagnosis of PVT between January 2018 and December 2022. We assessed the rate of and risk factors for PC. In total, 189 patients with PVT were included; the rate of PC was 14.8%. In univariate and multivariate analysis, the main risk factors for the presence of PC were etiology (thrombophilia, myeloproliferative disorders, local inflammatory diseases, and idiopathic causes), prior PVT, and complete versus incomplete or single-branch portal obstruction. In patients with superior mesenteric vein (SMV) thrombosis, distal obstruction was more prone to PC than proximal obstruction. The main predictive factors were etiology, prior PVT, complete PVT obstruction, and no prior non-selective beta-blocker (NSBB) use; in patients with SMV thrombosis, the distal extension was more significantly associated with the risk of PC. We propose a composite score for the prediction of PC which includes etiology, prior diagnosis of PVT, prior NSBB use, complete versus incomplete PVT, and distal versus proximal SMV thrombosis, with good accuracy (AUC 0.822) and an estimated sensitivity of 76.92% and specificity of 82.39% at a cut-off value of 4. Full article

Dynamic perviousness is a novel imaging biomarker, with clot density measurements at multiple timepoints to allow longer contrast to thrombus interaction. We investigated the correlations between dynamic perviousness and clot composition in the setting of acute ischemic stroke. Thirty-nine patients with large vessel occlusion (LVO) undergoing mechanical thrombectomy (MT) were analyzed. Patients received a three-phase CT imaging pre-thrombectomy and histopathological analysis of retrieved clots. Clot densities for every phase and change in densities between phases were calculated, leading to four patterns of dynamic perviousness: no contrast uptake, early contrast uptake with and without washout and late uptake. Clots were categorized into three groups based on dominant histologic composition: red blood cell (RBC)-rich, fibrin/platelet-rich and mixed. Clot composition was correlated with dynamic perviousness using the Kruskal–Wallis test and Pearson’s correlation analysis. The dynamic perviousness categories showed a significant difference between fibrin-rich clots when compared to RBC-rich plus mixed groups. The uptake without washout category had significantly fewer fibrin clots compared to the uptake with washout (p = 0.036), and nearly significantly fewer fibrin clots when compared to the no uptake category (p = 0.057). Contrast uptake with different patterns of contrast washout showed significant differences of the likelihood for fibrin-rich clots. Full article

This article presents the biochemical properties of poly(lactide)-zinc (PLA-Zn) composites obtained by DC magnetron sputtering of zinc onto melt-blown nonwoven fabrics. The biochemical properties were determined by the evaluation of the activated partial thromboplastin time (aPTT) and prothrombin time (PT). The antimicrobial activity of the PLA-Zn samples was additionally tested against representative Gram-positive and Gram-negative bacteria strains. A structural study of the PLA-Zn has been carried out using specific surface area and total pore volume (BET) analysis, as well as atomic absorption spectrometry with flame excitation (FAAS). PLA-Zn composites exhibited an antibacterial effect against the analyzed strains and produced inhibition zones against E. coli and S. aureus. Biochemical investigations revealed that the untreated PLA fibers caused the acceleration of the clotting of human blood plasma in the intrinsic pathway. However, the PLA-Zn composites demonstrated significantly different properties in this regard, the aPTT was prolonged while the PT was not altered. Full article

This study aimed to compare the chemical composition of aqueous extracts from different aerial plant parts (leaves, stems, and flowers) of Onopordum nervosum ssp. platylepis growing in different regions (Sousse, Kairouan, and Nabeul) in Tunisia, as well as their antioxidant and milk-clotting properties for both spontaneously grown and cultivated plants. Results showed that phenolic composition varies significantly among ecotypes and plant organs (p < 0.05), with flowers containing the highest amounts of total phenols and flavonoids in both plant types. The flowers from Nabeul (NA) region showed the highest amounts of total phenols and higher phenolic contents compared to leaves and stems, with 44.75 mg GAE/g and 39.79 mg GAE/g in spontaneous and cultivated plants, respectively. However, flowers of spontaneously plants grown in Sousse (SO) showed the highest total flavonoid contents (11.42 mg QE/g). Additionally, the findings indicated that flowers contained higher concentrations of mono- and disaccharides than leaves and stems. The antioxidant activity showed that the radical scavenging activity of O. platylepis aqueous extracts is significantly affected by the organ and genotype (p < 0.05). NA genotype revealed the highest potency in inhibiting free radicals, with flowers having the lowest IC50s values in 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) tests, registering 0.13 ± 0.02 and 0.14 ± 0.01 mg/mL, respectively. Milk-clotting activity was limited to flowers of this thistle. Moreover, SO and NA genotypes were identified as the most potent populations to coagulate milk in spontaneous and cultivated plants. The principal component analysis confirms the organ and genotype variability in this thistle, with the first two axes explaining 64.15% of the variance and highlighting a distinct flower group. The obtained results suggest that the domestication of this thistle could be useful for the conservation of biodiversity and the promotion of genotypes with potential technological properties. Full article

Although fibrin matrices derived from Platelet-Rich Plasma (PRP) are widely used in regenerative medicine, they have some limitations that can hinder their application. Modifying the composition of the PRP-derived fibrin matrix may improve its properties, making it suitable for certain medical uses. Three types of fibrin matrices were obtained: a PRP-derived fibrin matrix (FM), a PRP-derived fibrin matrix with a high fibrinogen content and platelets (FM-HFP) and a PRP-derived fibrin matrix with a high fibrinogen content (FM-HF). The fibrinogen levels, biomechanical properties and cell behavior were analyzed. The presence of platelets in the FM-HFP generated an inconsistent fibrin matrix that was discarded for the rest of the analysis. The fibrinogen levels in the FM-FH were higher than those in the FM (p < 0.0001), with a concentration factor of 6.86 ± 1.81. The values of clotting and swelling achieved using the FM-HF were higher (p < 0.0001), with less clot shrinkage (p < 0.0001). The FM had a significantly higher stiffness and turned out to be the most adherent composition (p = 0.027). In terms of cell viability, the FM-HF showed less cell proliferation but higher live/dead ratio values (p < 0.01). The increased fibrinogen and platelet removal in the FM-HF improved its adhesion and other biomechanical properties without affecting cell viability. Full article