Search Results (50)

The pisco industry generates significant environmental waste, particularly grape pomace, which is a rich source of phenolic compounds. Emerging extraction technologies offer promising alternatives for recovering these bioactive components. This study evaluated enzyme-assisted extraction (EAE) and pressurized liquid extraction (PLE) techniques using response surface methodology to optimize phenolic compound yield and antioxidant capacity. Specifically, a D-optimal design was applied for EAE, and a Box–Behnken design was applied for PLE. The optimal extraction conditions for EAE were 0.75 U/mL of tannase, 40 U/mL of cellulase, 20 °C, and 15 min. For PLE, the optimal parameters were 54% ethanol, 113 °C, and three extraction cycles. These conditions yielded 38.49 mg GAE g⁻¹ dw and 50.03 mg GAE g⁻¹ dw of total polyphenols and antioxidant capacities of 342.47 μmol TE g⁻¹ dw and 371.00 μmol TE g⁻¹ dw, respectively. The extracts obtained under optimal conditions were further characterized through chromatographic techniques to determine their phenolic profiles. Seven phenolic compounds were identified: gallic acid, catechin, epicatechin, 4-hydroxybenzoic acid, quercetin-3-rutinoside hydrate, quercetin-3-O-rhamnoside, and kaempferol. PLE extracts exhibited the highest concentration of these compounds. These findings demonstrate that recovering antioxidant-rich phenolic compounds from pisco grape pomace using innovative extraction methods is a viable strategy for obtaining functional ingredients and supporting sustainable industrial practices. Full article

Background/Objectives: Patients diagnosed with decompensated congestive heart failure (HF) often have elevated CA-125 levels, attributed to systemic congestion. However, a subgroup of patients presents with normal CA-125 levels. The primary objective of this study was to characterize the clinical, analytical, and echocardiographic profiles of patients admitted for decompensated congestive HF according to their CA-125 levels. The secondary objective was to analyze mortality after discharge. Methods: We conducted a retrospective study of patients hospitalized for a decompensated congestive HF episode. Recruitment was consecutive over more than 4 years (December 2019–June 2024), with 3151 patients recruited. Scheduled admissions, transfers from other hospitals, pulmonary congestion patterns, mixed patterns, and low output were the exclusion criteria. The final number of patients included was 166, all with an isolated systemic congestion pattern: CA-125 ≤ 50 U/mL: 38, and CA-125 > 50 U/mL: 128. Results: The comparative analysis between the groups showed that patients with CA-125 ≤ 50 U/mL were more often women (p < 0.05). They also had lower bilirubin and GOT/AST levels (p < 0.05). The percentage of patients with a preserved left ventricular ejection fraction (≥50%) was higher in the CA-125 ≤ 50 U/mL group (p < 0.05). The right ventricular (RV) size and inferior vena cava (IVC) were enlarged in both groups but with no significant differences (p < 0.05). However, the degree of RV dysfunction was greater in the CA-125 > 50 U/mL group, while the proportion of patients with inspiratory collapse of the IVC was higher in the CA-125 ≤ 50 U/mL group (p < 0.05). Survival curves differed from the first month and throughout the follow-up, with higher mortality in the CA-125 > 50 U/mL group. Thus, the probability of being alive at the end of the follow-up was over 50% in the CA-125 ≤ 50 U/mL group, while in the CA-125 > 50 U/mL group, it was around 25% (p < 0.05). Conclusions: The proportion of patients with decompensated congestive HF and systemic congestion who present with a low CA-125 level is close to 25%. These patients are mostly women with a preserved ejection fraction and inspiratory collapse of the IVC of >50%. Moreover, they have a higher survival rate, so a low CA-125 could help identify a subgroup of patients with a better prognosis. Full article

This study investigated the impact of enzyme treatment on the physicochemical parameters and volatile and bioactive composition of Arabica coffee beans during self-induced anaerobic fermentation (SIAF). The physicochemical parameters of the beans treated with the enzyme solution were monitored over a 120 h fermentation period. The results showed that increasing enzyme concentration reduced the levels of reducing sugars and phenolic compounds, leading to decrease in antioxidant activity. Pectin lyase activity was highest in beans treated with 10 U.·mL⁻¹, while polygalacturonase activity fluctuated throughout fermentation. The highest caffeine content (722.09 ± 3.7 mg·100g⁻¹) was found in beans treated with 5 U.mL⁻¹ after 72 h of fermentation. In contrast, trigonelline (1028.75 ± 31.4 mg·100g⁻¹) and 5-O-caffeoylquinic acid (5CQA) (423.46 ± 40.3 mg·100g⁻¹) were more prominent in unfermented beans. Volatile formation showed a positive correlation with enzyme concentration, with beans treated with 10 U·mL⁻¹ exhibiting a more diverse volatile profile in the first 24 h. These findings suggest that enzymatic treatment modulates coffee’s volatile and bioactive composition, enhancing levels of aromatic compounds that are directly linked to the sensory quality of the coffee beverage. Full article

Chitin deacetylase (CDA) plays a pivotal role in converting chitin to chitosan, yet industrial applications remain constrained by low enzymatic activity, instability under process conditions, and insufficient understanding of metalloenzyme activation mechanisms. Addressing these challenges, we conducted a genome-driven investigation of 151 salt-tolerant Bacillus strains to identify robust CDAs tailored for industrial demands. Genomic analysis revealed 120 strains harboring CDA genes, with Bacillus pumilus B866 exhibiting the highest native activity (105.93 U/mL). Through systematic medium optimization—identifying lactose, yeast extract, and FeSO₄ as critical components—CDA production in B866 surged to 191.32 U/mL, a 2.39-fold increase over baseline. Heterologous expression of BpCDA in E. coli yielded a recombinant enzyme (123.27 U/mL) with superior thermostability (retaining > 42.9% activity after 24 h at 55 °C) and broad pH adaptability (>81.4% activity at pH 7.0–9.0). Notably, BpCDA demonstrated unique Fe²⁺-dependent activation (186.4% activity enhancement at 1 mM), contrasting with Mg²⁺-dependent systems in prior studies. Comparative genomic and pan-genome analyses underscored evolutionary adaptations linked to saline–alkaline niches, while biosynthetic gene cluster profiling revealed strain-specific metabolic potentials independent of genome size. This study resolves critical limitations in CDA performance by integrating genome mining, targeted screening, and metalloenzyme engineering, establishing a scalable platform for sustainable chitin valorization. The optimized BpCDA, with its industrial-compatible stability and novel activation mechanism, represents a significant advancement toward efficient, eco-friendly chitosan production. Full article

Despite significant efforts to promote web accessibility through the adoption of various standards and tools, the web remains inaccessible to many users. One of the main barriers is the limited knowledge of accessibility issues among website designers. This gap in expertise results in the development of websites that fail to meet accessibility standards, hindering access for people with diverse abilities and needs. In response to this challenge, this paper presents the ACG WebAcc prototype, which enables the automatic generation of accessible HTML code using a model-driven development (MDD) approach. The tool takes as input a Unified Modeling Language (UML) model, with a specific profile, and incorporates predefined Object Constraint Language (OCL) rules to ensure compliance with accessibility guidelines. By automating this process, ACG WebAcc reduces the need for extensive knowledge of accessibility standards, making it easier for designers to create accessible websites. Full article

L-asparaginase (L-ASNase) functions as a chemotherapeutic enzyme with antitumor properties. It facilitates the degradation of L-asparagine (L-ASN), a vital amino acid required for the proliferation of tumor cells. In this study, we have isolated 177 L-ASNase-producing strains from the aquatic environment of the Arabian–Persian Gulf. The most potent isolate, ASP-J1-4, was an endophyte recovered from the seablite Suaeda maritima and was molecularly identified as B. xiamenensis (accession number PQ593941). The enzyme purified through DEAE-Sepharose displayed a molecular weight of 37 kDa based on the SDS-PAGE profile and lacked detectable L-glutaminase (L-GTNase) activity. Optimal enzyme activity was at 40 °C and pH 9.0, with stability at pH 7–9. The maximum stimulation effect was found in the presence of Fe³⁺, Mn²⁺, and Na⁺ ions, respectively. The enzyme demonstrated a V_max of 35.71 U/mL and a K_m of 0.15 mM. Interestingly, ASP-J1-4 L-ASNase showed a dose-dependent inhibition against human colon carcinoma (HCT-116) and cervical Henrietta Lacks (HeLa) cell lines, with IC₅₀ values of 15.42 µg/mL and 12.13 µg/mL, respectively. These findings collectively suggest a biocompatible, efficient, and robust enzyme for potential applications in tumor therapy after validation of in vivo studies and clinical trials. This study introduces the first deep screening program for L-ASNase-producing bacteria harboring in the Arabian–Persian Gulf region. In addition, it launches B. xiamenensis and other species as new sources of L-ASNase. Full article

Beer, the most popular alcoholic beverage, poses health risks for individuals with gout and hyperuricemia due to its high purine content. Herein, we identified a novel purine nucleoside phosphorylase (AbPNP) from the edible mushroom Agaricus bisporus and heterologously expressed it in Pichia pastoris. The recombinant AbPNP exhibited optimal activity at 60 °C and pH 7.0, retaining >80% activity at pH 6.0–9.0 and >85% activity after 3 h at ≤60 °C. Kinetic analysis revealed high catalytic efficiency (kcat/Km = 2.02 × 10⁶ s⁻¹⋅M⁻¹) toward inosine, with strong resistance to metal ions except for Co²⁺ and Cu²⁺. The application of AbPNP (1.0–5.0 U/mL) during wort saccharification reduced purine nucleosides by 33.54% (from 151.53 to 100.65 mg/L) while increasing yeast utilization of free purine bases. The resulting beer showed improved fermentation performance (alcohol content increased by 3.6%) without compromising flavor profiles. This study provides the food-grade enzymatic strategy for low-purine beer production, leveraging the GRAS status of both A. bisporus and P. pastoris. Full article

Introduction: Hepatitis D virus (HDV) infection remains a significant global health challenge due to its severity and high risk of progression to cirrhosis and hepatocellular carcinoma (HCC). Bulevirtide, a novel HDV entry inhibitor, has shown promise in managing chronic hepatitis D by blocking viral entry into hepatocytes. This study evaluated the efficacy and safety of bulevirtide in reducing HDV RNA levels and improving liver function in a real-life cohort of Italian patients with HDV infection. Methods: This multicenter prospective trial enrolled 108 consecutive patients with chronic HDV infection, from June 2023 to June 2024, who received 2 mg/day of bulevirtide in combination with a nucleoside/nucleotide analogue for hepatitis B virus (HBV) infection. Patients with any stage of liver fibrosis or compensated cirrhosis were included. Data collected included demographic and clinical characteristics, liver function tests, HDV RNA levels, and adverse events at baseline and 6 months. Results: The virological response was achieved in 54.6% of patients (n = 59), with 36 demonstrating undetectable HDV RNA levels. Among responders, ALT levels decreased significantly from 67.0 U/mL [IQR 44.0–116.3] to 31.5 U/mL [IQR 24.0–36.5, p = 0.001], and AST levels from 66.0 U/mL [IQR 46.5–91.0] to 32.5 U/mL [IQR 28.0–38.0, p = 0.021]. Median HDV RNA dropped from 29,800 IU/mL [IQR 3100–375,000] to 0 IU/mL [IQR 0–291, p < 0.001]. No significant predictors of response emerged. Mild adverse events, including pruritus (5.6%) and injection-site reactions (1.9%) and flu-like syndrome (0.9) were reported, with no treatment discontinuation. Conclusions: Bulevirtide effectively reduces HDV RNA levels and improves liver function with a favorable safety profile, offering a promising therapeutic option for chronic hepatitis D. Further large-scale studies are needed to confirm these findings and explore long-term outcomes. Full article

The Fusarium oxysporum species commonly found in soil include plant and human pathogens, and nonpathogenic species. F. oxysporum grown on lignocellulosic substrates under submerged conditions produces an extracellular enzyme profile with hemicellulolytic and cellulolytic activities. Our aim was to produce an extract of Fusarium oxysporum f. sp. melonis with β-1,4-xylanase activity after fermentation on a Brewers’ spent grain (BSG)-containing substrate. We prepared the BSG substrate, with or without sonication, for the submerged fermentation of Fusarium oxysporum previously isolated from local soil and preserved at 4 °C. First, an enriched inoculum was prepared, and later, the production of β-1,4-xylanase using the BSG substrates was monitored for up to 6 or 10 days in the enriched inoculum or in the enzyme extract, respectively. An activity of β-1,4-xylanase 12.0 U/mL (day 3) was obtained in the enriched inoculum with the untreated BSG, remaining constant for 3 days. A significant increase in the activity of this enzyme was observed (day 6), especially in the extract obtained using the sonicated BSG substrate (39 U/mL). Applying ultrasound to the BSG before its use in a submerged fermentation with Fusarium oxysporum f. sp. melonis could be an alternative for producing β-1,4-xylanase. Full article

Citric acid by-products in animal feed pose a sustainability challenge. Bacillus species are commonly used for fermenting and improving the nutritional quality of feedstuffs or by-products. An experiment was conducted to enhance the nutritional value of citric acid by-products through fermentation with Bacillus subtilis I9 for animal feed. The experiment was carried out in 500 mL Erlenmeyer flasks with 50 g of substrate and 200 mL of sterile water. Groups were either uninoculated or inoculated with B. subtilis I9 at 10⁷ CFU/mL. Incubation occurred at 37 °C with automatic shaking at 150 rpm under aerobic conditions for 0, 24, 48, 72, and 96 h. Inoculation with B. subtilis I9 significantly increased Bacillus density to 9.3 log CFU/mL at 24 h (p < 0.05). CMCase activity gradually increased, reaching a maximum of 9.77 U/mL at 72 h. After 96 h of fermentation with inoculated B. subtilis I9, the citric acid by-product exhibited a significant decrease (p < 0.05) in crude fiber by 10.86%, hemicellulose by 20.23%, and cellulose by 5.98%, but an increase in crude protein by 21.89%. Gross energy decreased by 4% after inoculation with B. subtilis in comparison to the uninoculated control (p < 0.05). Additionally, the non-starch polysaccharide (NSP) degradation due to inoculation with B. subtilis I9 significantly reduced (p < 0.05) NSP by 24.37%, while galactose, glucose, and uronic acid decreased by 22.53%, 32.21%, and 18.11%, respectively. Amino acid profile content increased significantly by more than 12% (p < 0.05), including indispensable amino acids such as histidine, isoleucine, lysine, methionine, phenylalanine, tryptophan, and valine and dispensable amino acids like alanine, aspartic acid, glutamic acid, glutamine, glycine, proline, and tyrosine. Furthermore, citric acid by-products inoculated with B. subtilis I9 exhibited changes in the cell wall structure under scanning electron microscopy, including fragmentation and cracking. These results suggest that fermenting citric acid by-products with B. subtilis I9 effectively reduces dietary fiber content and improves the nutritional characteristics of citric acid by-products for use in animal feed. Full article

The growing global market of dairy products has led to the need for alternative approaches regarding whey valorization, which is the primary by-product of cheese and strained yogurt production. In this context, prebiotic galactooligosaccharides can be produced enzymatically from whey using commercially available β-galactosidases. A comparative study was conducted to assess the production of galactooligosaccharides from sweet and acid whey, thereby employing two commercial β-galactosidases from Aspergillus oryzae and Kluyveromyces lactis. The study considered the initial lactose content and enzyme load as variables. The maximum yields of galactooligosaccharides in concentrated sweet whey (15% w/v initial lactose) and raw acid whey (3.1% w/v initial lactose) reached 34.4 and 14.7% with lactase from Kluyveromyces lactis (0.13 U/mL), respectively. The corresponding galactooligosaccharide yields for lactase from Aspergillus oryzae were equal to 27.4 and 24.8% in the most concentrated sweet and acid whey, respectively, using enzyme loads of 2 U/mL in sweet whey and 1 U/mL in acid whey. Concerning the profile of the produced galactooligosaccharides, the Kluyveromyces lactis lactase hydrolyzed lactose more rapidly and resulted in higher levels of allolactose and lower levels of 6-galactosyl-lactose, compared to the lactase from Aspergillus oryzae, and achieved in both cases a polymerization degree of up to six. Full article

Xylanases are hydrolytic enzymes that have tremendous applications in different sectors of life, but the high cost of their production has limited their use. One solution to reduce costs and enhance xylanase production is the use of agro-wastes as a substrate in fungal cultures. In this study, olive mill pomace (OMP) and barley bran (BB) were used as carbon sources and possible inducers of xylanase production by three species of Trichoderma (atroviride, harzianum, and longibrachiatum), one major xylanase producer. The experiments were conducted under a solid-state fermentation system (SSF) in flask cultures and a packed-bed bioreactor. Cultures of OMP and BB were optimized by examining different ratios of OMP and BB, varied particle sizes, and inoculum size for the three species of Trichoderma. The ratio of 8:2 OMP and BB yielded the highest xylanase activity, with a particle size of 1 mm at 29 °C and an inoculum size of 1 × 10⁷ spores/mL. Studying the time profile of the process revealed that xylanase activity was highest after seven days of incubation in flask SSF cultures (1.779 U/mL) and after three days in a packed-bed bioreactor (1.828 U/mL). The maximum percentage of OMP degradation recorded was about 15% in the cultures of T. harzianum flask SSF cultures, compared to about 11% in T. longibrachiatum bioreactor cultures. Ammonium sulfate precipitation and dialysis experiments showed that Xylane enzyme activity ranged from 0.274 U/mL in T. harzianum to 0.837 U/mL in T. atroviride when crude extract was used, with the highest activity (0.628 U/mL) at 60% saturation. Xylose was the main sugar released in all purified fractions, with the G-50 and G-75 fractions showing the maximum units of xylanase. Full article

This study investigated the effect of red corn in the feed mixture of dairy goats on production traits, blood metabolic profile, and antioxidative status. The study was conducted on 30 French Alpine dairy goats. The feed mixture for the goats in the control group (CC) contained yellow corn (100%). In the first experimental group (RC50), yellow corn was partially (50%) replaced by red corn (RC), and in the second experimental group (RC100), yellow corn was completely (100%) replaced by red corn. No significance variations (p > 0.05) were determined in production traits of dairy goats between dietary treatments. A significant increase in hemoglobin (84.43, 100.00 and 106.55 g/L), mean corpuscular hemoglobin (7.98, 9.70 and 12.54 pg), and mean corpuscular hemoglobin concentration (293.57, 357.50 and 462.78 g/L) was found in the RC groups, and a decrease in erythrocytes in the RC100 compared with the RC50 group of goats (from 8.71 to 10.45 × 10¹² L). A significant increase in blood superoxide dismutase (SOD) activity in the RC groups was found (0.29, 0.53, and 0.44 U/mL). The results indicate maintaining production traits and a moderate effect on blood metabolic profile (most hematologic parameters) as well as a positive antioxidative effect RC. Full article

Background and Objectives: Psoriasis is an immune-mediated chronic inflammatory skin disorder and commonly associated with highly noticeable erythematous, thickened and scaly plaques. Deubiquitinase genes, such as tumor necrosis factor-alpha protein 3 (TNFAIP3, A20), the cylindromatosis (CYLD) and Cezanne, function as negative regulators of inflammatory response through the Janus kinase/signal transducers and activators of transcription (JAK-STAT) pathways. In this study, polymorphisms and expressions of A20, CYLD and Cezanne genes as well as immunophenotype in psoriatic patients were determined. Materials and Methods: In total, 82 patients with psoriasis and 147 healthy individuals with well-characterized clinical profiles were enrolled. Gene polymorphisms were determined by direct DNA sequencing, gene expression profile by quantitative real time-polymerase chain reaction (PCR), immunophenotype by flow cytometry, and the secretion of cytokines and cancer antigen (CA) 125 by enzyme-linked Immunosorbent assay (ELISA). Results: The inactivation of A20, CYLD and Cezanne and increased levels of TNF-α, IFN-γ and CA 125 was observed in psoriatic patients. Importantly, patients with low A20 expression had significant elevations of triglyceride and total cholesterol concentrations and higher numbers of CD13⁺CD117⁻ and CD19⁺CD23⁺ (activated B) cells than those with high A20 expression. Genetic analysis indicated that all rs4495487 SNPs in the JAK2 gene, rs200878487 SNPs in the A20 gene and four SNPs (c.1584-375, c.1584-374, rs1230581026 and p.W433R) in the Cezanne gene were associated with significant risks, while the rs10974947 variant in the JAK2 gene was at reduced risk of psoriasis. Moreover, in the Cezanne gene, p.W433R was predicted to be probably damaging by the Polyphen-2 prediction tool and an AA/CC haplotype was associated with a high risk of psoriasis. In addition, patients with higher CA 125 levels than the clinical cutoff 35 U/mL showed increased levels of IFN-γ than those with normal CA 125 levels. Conclusions: A20 expression was associated with lipid metabolism and the recruitment of CD13⁺ CD117⁻ and activated B cells into circulation in psoriatic patients. Besides this, the deleterious effect of the p.W433R variant in the Cezanne gene may contribute to the risk of psoriasis. Full article

The paper analyzes quasiperiodic upwellings and downwellings on the shelf and upper part of continental slope of the northeastern Black Sea. It is shown that these processes are related to changes in intensity and direction of alongshore current and the following geostrophic adjustment of the density field. The source of such changes is the meandering of the Black Sea Rim Current (RC). It leads to a quasiperiodic change in direction of the alongshore current, from northwestern (cyclonic RC meander) to southeastern (anticyclonic RC meander, or eddy). These cycles, or phases, have an average duration of about 10 days. During the northwestern phase, the permanent Black Sea pycnohalocline (hereafter pycnocline) and seasonal thermocline descend, their thickness increases, and so does the thickness of the upper mixed layer (UML). During the southeastern phase, both the pycnocline and seasonal thermocline ascend and become thinner, along with the UML, which also becomes thinner. In both phases, isopycnals in the pycnocline and isotherms in the thermocline demonstrate quasi-in-phase vertical oscillations, which have a good correlation with the speed and direction of the alongshore current. These correlations allow estimation of the magnitude of upwellings and downwellings in the shelf–slope area of the northeastern Black Sea using data series of current velocity profiles. Full article