Protein Concentration Research Articles

A single-particle analysis method for detecting membrane remodelling and curvature sensing.

In biology, shape and function are related. Therefore, it is important to understand how membrane shape is generated, stabilised and sensed by proteins and how this relates to organelle function. Here, we present an assay that can detect curvature preference and membrane remodelling with free-floating liposomes using protein concentrations in physiologically relevant ranges. The assay reproduced known curvature preferences of BAR domains and allowed the discovery of high-curvature preference for the PH domain of AKT and the FYVE domain of HRS (also known as HGS). In addition, our method reproduced the membrane vesiculation activity of the ENTH domain of epsin-1 (EPN1) and showed similar activity for the ANTH domains of PiCALM and Hip1R. Finally, we found that the curvature sensitivity of the N-BAR domain of endophilin inversely correlates to membrane charge and that deletion of its N-terminal amphipathic helix increased its curvature specificity. Thus, our method is a generally applicable qualitative method for assessing membrane curvature sensing and remodelling by proteins.

Deciphering the mechanism underlying poor aqueous solubility of extracted quinoa proteins

This study aimed to decipher the mechanisms underlying poor solubility of quinoa proteins by investigating the form of quinoa proteins dispersed in water and how protein-protein interactions influenced the kinetic stability of proteins in the dispersions. Specifically, the relative solubility and the forms of quinoa proteins in 1–5 w/w% protein dispersions were determined by separating proteins via centrifugation and/or ultrafiltration. The kinetic stability of quinoa proteins in the supernatants over a 3-week storage period was characterized by determining the changes of concentration, composition and physicochemical properties of quinoa proteins and predicting protein-protein interactions. The results showed that quinoa proteins existed mainly as differently-sized protein aggregates in the dispersions, leading to low relative solubility. The coagulation of protein aggregates in the supernatants caused severe precipitation during the first week of storage whereas they were disassociated simultaneously. With further storage, the remaining proteins in the supernatants reached kinetic stability, which was contributed by stronger electrostatic repulsion and lower surface hydrophobicity. Moreover, 11S globulin and 2S albumin were precipitated and solubilized together during storage, which was ascribed to intermolecular interactions driven by multiple sites between 11S globulin and/or 2S albumin. This study lays a foundation for extensive utilization of quinoa proteins.

Essential oils cause membrane disruption and autoaggregation of MDR Acinetobacter baumannii cells

Essential oils are promising antimicrobial agents against various bacteria. The aim of the study was to examine anti-Acinetobacter baumannii activity of nine essential oils and to elucidate essential oil mechanisms of action against this. In total, 8 out of 9 examined different essential oils exhibited activity against multidrug-resistant (MDR) A. baumannii isolates. The best anti-A. baumannii activity expressed Thymus serpyllum, Satureja hortensis and Oreganum majorana essential oil (MBC 1.29 to 2.58 µg mL−1), while essential oils of Menta x piperita, Hyssopus officinalis, Foeniculum vulgare and Artemisia dracunculus expressed the same effect in higher concentrations (1.80 to 3.80 µg mL−1). The effect of Juniperus sabina, Juniperus sibirica and S. hortensis essential oils against A. baumannii is for the first time reported here. The mechanisms of Myrtus communis, Eucalyptus camaldulensis, S. hortensis and T. serpyllum essential oils were further examined. The essential oil treatments led to concentration dependent leakage of biomolecules form the cells, causing an increase in the concentrations of lipids, carbohydrates, and proteins in the suspensions. Microstructural observations confirmed the essential oil effect on cell membranes and disruption the membrane integrity, with obvious leakage of intercellular substance, collapsed cells with perforations, debris presence and autoaggregated cells. The subinhibitory concentrations of oils did not obviously changed protein patterns determined by SDS-PAGE. These results indicate that the essential oils, particularly T. serpyllum, S. hortensis, M. communis and E. camaldulensis express its anti-A. baumannii activity via membrane disruption and increased membrane permeability, representing very promising alternative antibacterial agents against MDR wound isolates.

Complement proteins and complement regulatory proteins are associated with age-related macular degeneration stage and treatment response

BackgroundDysregulation of the complement system is involved in development of age-related macular degeneration (AMD). The complement cascade is regulated by membrane bound complement regulatory proteins (Cregs) on mononuclear leukocytes among others. This study aims to investigate systemic complement proteins and Cregs in AMD stages and their association with treatment response in neovascular AMD (nAMD).MethodsIn this clinical prospective study, treatment-naïve patients with nAMD, intermediate AMD (iAMD) and healthy controls were recruited and systemic complement proteins C3, C3a and C5a were investigated with electrochemiluminescence immunoassays, and Creg expression (CD35, CD46 and CD59) on T cells (CD4 + and CD8+) and monocytes (classical, intermediate and non-classical) investigated with flow cytometry. Treatment response in nAMD patients was evaluated after loading dose and after one year, and categorized as good, partial or poor. Complement proteins and Creg expression levels were compared between healthy controls, iAMD and nAMD, as well as between good, partial and poor nAMD treatment response groups. Polymorphisms in the CFH and ARMS2 genes were analyzed and compared to complement proteins and Creg expression levels in nAMD patients.ResultsOne hundred patients with nAMD, 34 patients with iAMD and 61 healthy controls were included. 94 nAMD patients completed the 1-year follow-up. Distribution of treatment response in nAMD was 61 (65%) good, 26 (28%) partial, and 7 (7%) poor responders. The distribution of 1-year treatment response was 50 (53%) good, 33 (36%) partial, and 11 (11%) poor responders. The concentrations of systemic C3, C3a, and the C3a/C3-ratio were significantly increased in patients with nAMD compared to healthy controls (P < 0.001, P = 0.002, and P = 0.035, respectively). Systemic C3 was also increased in iAMD compared to healthy controls (P = 0.031). The proportion of CD46 + CD4 + T cells and CD59 + intermediate monocytes were significantly decreased in patients with nAMD compared to healthy controls (P = 0.018 and P = 0.042, respectively). The post-loading dose partial treatment response group had significantly lower concentrations of C3a and C5a compared to the good response group (P = 0.005 and P = 0.042, respectively). The proportion of CD35 + monocytes was significantly lower in the 1-year partial response group compared to the 1-year good response group (P = 0.039). High-risk CFH genotypes in nAMD patients was associated with increased C3a, C3a/C3-ratio, and expression levels of CD35 + CD8 + T cells and CD46 + classical monocytes, while expression level of CD46 + non-classical monocytes was decreased.ConclusionElevated concentrations of systemic complement proteins were found in patients with iAMD and nAMD. Decreased Creg expression levels were found in patients with nAMD. Partially responding nAMD patients had a dysregulated complement system and Cregs compared to good responders.

Assessment of clinical and laboratory parameters of oral сavity in patients with orthodontic systems

equipment (braces) on the biochemical parameters of oral fluid. Materials and methods. The study involved patients aged 18 to 22 years who are undergoing orthodontic treatment with a bracket system. Thegroup of examined persons was 25 people. The control group included 15 practically healthy people without braces. All patients underwent dental status assessment (Green – Vermillion hygiene index – simplified IG-Y; PMA inflammation index), laboratory examination of oral fluid before and after controlled oral hygiene. The biochemical analysis included determination of the acid-base balance in the oral fluid using a pH meter, assessment of free radical oxidation by chemiluminogram parameters by biochemiluminescence and by levels of molecular products of lipid peroxidation (diene and triene conjugates, Schiff bases), activity of the enzyme alkaline phosphatase, concentrations of total protein and lactate. Results and discussions. In patients undergoing orthodontic treatment with a bracket system, an insufficient level of oral hygiene and inflammatory manifestations in the gums of moderate severity were revealed. The level of the acid-base balance of the oral fluid before controlled hygiene was equal to 6.8±0.19, that is, below the range of normal pH values in the oral cavity. After hygiene, the pH of the oral fluid returned to the norm of 7.1±0.2. In patients with braces, activation of free radical oxidation and an increase in lipoperoxidation parameters – triene conjugates and Schiff bases, increased activity of alkaline phosphatase, total protein and lactate levels took place.Cоnclusion. After hygienic treatment of the oral cavity, the biochemical parameters did not differ statistically significantly from the control data.

Long-term monocyte activation after coronary artery bypass grafting: an exploratory prospective observational study

Major surgery such as coronary artery bypass grafting (CABG) is associated with an increased post-operative risk of atherosclerotic cardiovascular events. Cells of the innate immune system can adopt a long-lasting pro-inflammatory and atherogenic phenotype after brief exposure to exogenous or endogenous inflammatory stimuli, a process called “trained immunity”. We hypothesized that the surgery-induced inflammation leads to sustained alterations in monocyte function, which promote the subsequent occurrence of cardiovascular events. Blood from 13 patients undergoing elective CABG was obtained before, 3-7 days (median 4) after, and 6-8 weeks (median 6) weeks after surgery. At 3-7 days postoperatively, circulating C-reactive protein (CRP) concentration, leukocyte counts and ex vivo Peripheral Blood Mononuclear Cell (PBMC) IL-6, TNFα and IL-1Ra production after stimulation (with various inflammatory stimuli) were significantly increased. Simultaneously, there was a reduction in monocyte HLA-DR expression. 6-8 weeks after CABG there was an ongoing systemic pro-inflammatory state with higher CRP concentrations, increased stimulated ex vivo PBMC IL-6 production, changes in monocytes subsets, and a higher expression of CCR2 on monocytes compared to baseline. In conclusion, CABG induces a persistent systemic inflammatory reaction with a sustained activated monocyte phenotype. This might contribute to the increased atherosclerotic cardiovascular event risk observed in cardiac surgery patients.

Nutritional composition of black soldier fly (Hermetia ilucens) larvae reared on vegetable wastes: effects on growth of village chickens

Context Black soldier fly larvae are a promising alternative ingredient in poultry feed. They might be able to replace soybean and fishmeal because the have high concentrations of protein and fat, and are also rich in vitamins and minerals, and because partial replacement of soybean and total replacement of fishmeal produce no adverse effect on the growth performance of village chickens. Therefore, they are a suitable replacement for these traditional ingredients in poultry feed. Aims This study determines the nutritive values of black soldier fly larvae and evaluates the effects of feeding diets containing black soldier fly larvae on the growth performance of village chickens. Methods Samples of black soldier fly larvae were subjected to proximate analysis before a total of 280 1-day-old village chicks were randomly divided into four groups. All groups were fed a basal diet with partial replacements of corn and soybean with black soldier fly larvae at 0% (Control), 5% (T1), 10% (T2) and 15% (T3) for 70 days. Each treatment group consisted of seven replicates, with 10 birds per replicate. The bodyweights, bodyweight gains and feed conversion rates were determined. Key results Dry rendered black soldier fly larvae contained 773.0 ± 0.08 g/kg dry matter, 36.1 ± 0.09 g/kg ash, 408.8 ± 0.28 g/kg crude protein, 283.0 ± 0.16 g/kg ether extract and 40.9 ± 0.44 g/kg crude fibre. The larvae also contained 2.041 MJ/kg of metabolisable energy, 20.4 g/kg of calcium and adequate concentrations of vitamins. The feeding trial showed an average bodyweight gain of 1231.45 ± 23.37 g and feed conversion ratio of 2.03 ± 0.15, which were most efficient with Diet T3. Conclusions Inclusion of 15% black soldier fly larvae in diets can enhance the growth performance of village chickens without adverse effects. Implications Partial replacement of soybean and total replacement of fishmeal in the diet of chickens by black soldier fly larvae are not affecting the growth performance of village chickens but might reduce the feed cost.

Heterogeneous Slowdown of Dynamics in the Condensate of an Intrinsically Disordered Protein.

The high concentration of proteins and other biological macromolecules inside biomolecular condensates leads to dense and confined environments, which can affect the dynamic ensembles and the time scales of the conformational transitions. Here, we use atomistic molecular dynamics (MD) simulations of the intrinsically disordered low complexity domain (LCD) of the human fused in sarcoma (FUS) RNA-binding protein to study how self-crowding inside a condensate affects the dynamic motions of the protein. We found a heterogeneous retardation of the protein dynamics in the condensate with respect to the dilute phase, with large-amplitude motions being strongly slowed by up to 2 orders of magnitude, whereas small-scale motions, such as local backbone fluctuations and side-chain rotations, are less affected. The results support the notion of a liquid-like character of the condensates and show that different protein motions respond differently to the environment.

Impact of Storage Temperature on Physical Stability and Protein-Lipid Co-oxidation in Whey Protein functional Emulsions.

The aim of this study was to investigate the effect of different storage temperatures (4°C, 25°C and 45°C, denoted as T-4, T-25 and T-45, respectively) on the physical stability and protein-lipid co-oxidation of whey protein functional emulsions during the storage period at storage times of 1, 2, 3 and 6 months. At the 6th month of storage, samples stored at 45°C were less physically stable than samples stored at 4°C, as evidenced by an increase in particle size (5.50%), a decrease in zeta potential (7.93%), and a decrease in adsorbed protein concentration (36.8%). The most severe protein-lipid co-oxidation was induced in whey protein functional emulsions under 45°C temperature conditions with increasing storage time. Protein oxidation products (carbonyls, N′-formyl-L-kynurenine, and sulfhydryl groups) and lipid oxidation products (lipid hydroperoxides, thiobarbituric acid reactive substances) were at their highest levels, and significant molecular changes were observed in the molecules of adsorbed protein or unadsorbed proteins. As the storage temperature rises it exposes the active sites such as internal sulfhydryl and hydrophobic groups in the protein, promoting cross-linking and aggregation. Cluster and correlation analyses showed that protein-lipid co-oxidation of emulsions inevitably occurs during storage, but low-temperature storage can delay this process. This study is of guiding significance for the quality maintenance of functional nutritional emulsions and the selection of storage conditions. In addition, this study may provide clearer solutions to the problems of whey protein denaturation and formation of lipid oxides caused by protein-lipid co-oxidation in the food industry, as well as to the problem of increasing the shelf life of products by choosing the appropriate storage, packaging, and transportation temperatures.

Make it a standard? The creation and variability assessment of a consensus standard protocol for Tenebrio molitor larvae feeding trials

Abstract Interest in the nutrition of the yellow mealworm (Tenebrio molitor L.) larvae is on the rise, leading to an increase in publications on this topic. The absence of a standard protocol and resulting differences in experimental designs reduces comparability among studies and impedes research on mealworm nutrition. To address this, a consensus standardised protocol was developed specifically for the evaluation of mealworm larval growth and performance in feeding trials. The efficacy of this protocol was evaluated through an international ring test involving seven partners using two wheat brans as dry feed (a standard bran and a local bran) at 27 °C and 60% relative humidity. As experimental units, plastic crates filled with 2.1 kg of bran and 10,000 4-week-old larvae were used with six replicates. Agar gel was provided as wet feed ad libitum. The mean individual larval weight and the number of larvae per crate were determined weekly until either three or more replicates ran out of feed or pupation exceeded 10%. At harvest, the total larval fresh biomass and amount of frass was determined. Larval samples were taken for chemical analysis. To assess the protocol, the within (repeatability) and between (reproducibility) laboratory variability was calculated for each parameter. The repeatability was good (limit at 12% (standard) and 14% (local)). The reproducibility was poorer with a limit 2.7 times higher for the standard feed (36%) and 3.8 times higher for the local feed (55%). For both feeds, the total larval fresh harvest, amount of frass and the larval protein concentration were the most consistent both within and among laboratories. The highest variability was observed at the early life stages and for the larvae ash content. The detailed consensus standard protocol and repeatability/reproducibility estimates can be used as basis for future mealworm feeding trials, comparing results and future improvements.

Plasma proteomic signatures of liver steatosis and fibrosis in people living with HIV: a cross-sectional study

Insights into the mechanisms driving metabolic dysfunction-associated steatotic liver disease (MASLD) in people living with HIV (PLHIV) remain limited. Plasma proteomics holds promise for biomarker discovery and the elucidation of biological mechanisms. We performed cross-sectional analyses on data from 1036 virally suppressed PLHIV using antiretroviral treatment (ART) from the Dutch multi-centre 2000HIV cohort. Participants underwent transient elastography to assess liver steatosis (controlled attenuation parameter (CAP)≥263dB/m) and -fibrosis (liver stiffness measurement (LSM)≥7.0kPa). Plasma protein concentrations (n=2367) (Olink® Explore Panel) were compared between PLHIV with vs. without liver steatosis and PLHIV with vs. without fibrosis. Enriched pathways (using GO, KEGG and Reactome libraries) and correlations with clinical characteristics wereassessed, and analyses were stratified by BMI category. In addition, concentrations of 242 proteins werecompared between individuals ("controls") with and without liver steatosis (ratio of methylene:methylene and water >5.6% on magnetic resonance spectroscopy) from a separate cohort (300-OB), all having a BMI >26kg/m2. Steatosis and fibrosis were associated with 67/2367 (2.2%) and 17/2367 (0.7%) differentially expressed proteins (DEP), respectively, enriched in mostly metabolic pathways. Immunoglobulin superfamily member 9 (IGSF9) was amongst the top DEP associated with both steatosis and fibrosis. Stratifying by BMI revealed 8/2367 DEP associated with steatosis in lean- and 12/2367 DEP in overweight/obese individuals, with two shared DEP (IGSF9 and GHR). Conversely, protein signatures of overweight/obese PLHIV (32/242 DEP) and overweight/obese HIV-uninfected individuals (32/242 DEP) exhibited substantial overlap with 16 shared DEP. Notably, DEP correlated with HIV characteristics in lean individuals but not in overweight/obese PLHIV. Lean and overweight/obese PLHIV exhibit distinct proteomic signatures associated with liver steatosis, with the former being more strongly correlated with HIV-specific factors and ART. In addition, we identified a protein, IGSF9, strongly related to liver fibrosis and steatosis across BMI categories. The 2000HIV study is funded by ViiV Healthcare.

Phase separation drives the folding of recombinant collagen

Recombinantly produced collagens present a sustainable, ethical, and safe substitute for collagens derived from natural sources. However, controlling the folding of the recombinant collagens, crucial for replicating the mechanical properties of natural materials, remains a formidable task. Collagen-like proteins from willow sawfly are relatively small and contain no hydroxyprolines, presenting an attractive alternative to the large and post-translationally modified mammalian collagens. Utilizing CD spectroscopy and analytical ultracentrifugation, we demonstrate that recombinant willow sawfly collagen assembles into collagen triple helices in a concentration-dependent manner. Interestingly, we observed that the lower concentration threshold for the folding can be overcome by freezing or adding crowding agents. Microscopy data show that both freezing and the addition of crowding agents induce phase separation. We propose that the increase in local protein concentration during phase separation drives the nucleation-step of collagen folding. Finally, we show that freezing also induces the folding of recombinant human collagen fragments and accelerates the folding of natural bovine collagen, indicating the potential to apply phase separation as a universal mechanism to control the folding of recombinant collagens. We anticipate that the results provide a method to induce the nucleation of collagen folding without any requirements for genetic engineering or crosslinking.

Treatment with Leflunomide in Conjunction with Glucocorticoids for Dogs with Immune-Mediated Polyarthritis Is Not Associated with Improved Outcomes: A Retrospective Cohort Study of 93 Dogs from Australia (2017–2024)

Immune-mediated polyarthritis (IMPA) has a relatively high relapse rate compared to other immune-mediated diseases. Leflunomide is frequently used to treat dogs with IMPA in conjunction with prednisolone. This retrospective cohort study aimed to evaluate the therapeutic efficacy of leflunomide as an adjunctive therapy to prednisolone in reducing relapse and mortality rates in dogs diagnosed with IMPA in Australia. The medical records of client-owned dogs diagnosed with IMPA at a specialist referral hospital in Southeast Queensland from 2017 to 2024 were reviewed. A total of 93 dogs were included in this study, divided into two groups based on the treatment received: Group PRED, consisting of 53 dogs treated with prednisolone as the sole immunosuppressive agent, and Group L+PRED, consisting of 40 dogs that received leflunomide as adjunctive therapy alongside prednisolone. Data collected included breed, age, weight, sex, serum C-reactive protein concentration, results of synovial fluid analysis and microbial culture, treatment protocol, relapse rates and time to relapse, and mortality rates. There was no difference in relapse or mortality rates, time to relapse, nor time to discontinue prednisolone between the PRED and L+PRED groups. The L+PRED group had higher body weights and lower prednisolone dose rate at discharge compared to those in the PRED group. This study demonstrated that the use of leflunomide as an adjunctive therapy to prednisolone for the treatment of dogs with IMPA had no improved outcomes, reduced relapse rates, or shortening in the duration of prednisolone therapy when compared to dogs receiving prednisolone monotherapy.

Risk factors for and short-term prognosis of intrauterine infection in preterm infants

Objective To identify the risk factors for and short-term prognosis of intrauterine infection (IUI) in preterm infants. Methods We retrospectively collected clinical data regarding preterm infants (28+0–36+6 weeks of gestational age) with IUI from the neonatal intensive care unit of our hospital between June 2017 and June 2022. The pathologic characteristics of the infants were classified using the Redline criteria, and the resulting groups were compared with respect to their clinical characteristics, indicators of infection, complications, and mortality rates. Results Three hundred thirty (78.1%) of the 422 enrolled neonates showed signs of histologic IUI; 51.8% showed histologic chorioamnionitis alone, and 48.2% showed both histologic chorioamnionitis and funisitis. The independent risk factors identified for IUI were maternal prenatal invasive manipulation, premature rupture of membranes (&gt;18 hours before labor), and prenatal C-reactive protein concentration. The infants with IUI showed higher incidences of respiratory distress syndrome, bronchopulmonary dysplasia, retinopathy of prematurity, and prolonged mechanical ventilation. Low gestational age and low birth weight of preterm infants were significantly associated with a higher incidence of histologic IUI, more severe inflammation, and a higher incidence of complications. Conclusions Histologic IUI is associated with high complication and mortality rates in preterm infants, as is an increasing severity of inflammation.

Multiplex analysis of cancer cells treated with induced mesenchymal stem cell membrane vesicles

BACKGROUND: Extracellular vesicles (EVs) are membrane-derived vesicles released by cells into the extracellular space, playing a crucial role in intercellular communication and regulating a range of biological processes. These vesicles are found in tumor tissue, where they serve as mediators in signal transduction between tumor cells and the cells of the microenvironment. Similar to their parent mesenchymal stem cells (MSCs), extracellular vesicles demonstrate contradictory effects on tumor development. Studies have shown that MSC-derived EVs promote tumor growth; however, some research has also demonstrated their inhibitory role. AIM: The aim is to assess the effect of mesenchymal stem cell-derived membrane vesicles on the molecular composition of cancer cells. METHODS: Induced membrane vesicles were obtained from mesenchymal stem cells through treatment with cytochalasin B. Mesenchymal stem cells were initially sourced from adipose tissue. To simulate intercellular communication between tumor cells and MSCs, induced membrane vesicles were applied at varying protein concentrations to recipient cells (SH-SY5Y, PC3, MCF7). The bicinchoninic acid method was used to measure the total protein concentration isolated from human cells/induced membrane vesicles. Subsequently, the molecular composition of the recipient cells after the application of induced membrane vesicles was analyzed using multiplex assays. RESULTS: We determined that after the application of MSC-derived membrane vesicles to cancer cells, significant alterations occur in the expression of numerous biologically active molecules, including cytokines, chemokines, and growth factors. Specifically, increased concentrations of growth factor FGF-2, cytokines G-CSF, Fractalkine, IL-12p40, IL-9, IL-4, IL-6, IL-8, and chemokines IP-10, MCP-1, among others, were observed. The analysis also revealed that most of these molecules are associated with cell proliferation, migration, and immune response. CONCLUSION: Mesenchymal stem cell-derived membrane vesicles are capable of altering the molecular profile of cancer cells, increasing the concentration of molecules linked to cell survival and migration.

Physicochemical, techno-functional, and thermal properties of Kabuli chickpea's aquafaba extracted by optimized microwave-assistive process

Aquafaba, a liquid by-product from the soaking and cooking of legumes, is often discarded in canning processes despite its rich protein, carbohydrate content, and functional properties. This study investigates the impact of microwave irradiation pretreatment on the extraction efficiency and the physicochemical, techno-functional, and thermal properties of Kabuli chickpea aquafaba. The findings revealed that increased microwave irradiation time and power led to a reduction in foaming ability and extraction efficiency but significantly enhanced the soluble protein content, density, dry matter, and foam stability of aquafaba. Optimal extraction was achieved with 5 min of irradiation at 900 W, yielding the highest extraction efficiency (175.3 %), protein concentration (1779.12 mg kg-1), density (1.019 g/mL), dry matter (4.11 %), foaming ability (341.21 %), and foam stability (49.56 %). Thermogravimetric analysis and FT-IR spectroscopy confirmed that microwave pretreatment improved the thermal stability of aquafaba without altering its chemical structure. Microwave-assisted extraction improves the quality and functionality of aquafaba, offering a sustainable alternative to waste disposal in the food industry. While the initial investment in microwave technology may be high, it transforms aquafaba from a waste by-product into a valuable resource, leading to significant profitability for canning industries. Given the high techno-functional, physicochemical, and thermal properties of aquafaba, it appears to be a suitable alternative to more expensive ingredients like egg whites and a promising substitute in vegan products.

Synthesis of enzyme-enriched zinc oxide nanoparticles using Lantana camara L. fruit extract for detoxification of phenol and derivatives

In this study, we evaluated the potential of Lantana camara L. (L. camara L.) fruit extract as a biogenic mediator for the synthesis of enzyme-rich zinc oxide nanoparticles (EZnO) aimed at the detoxification of phenol and its derivatives. The protein extraction yielded a maximum concentration in the 80 % cut fraction, with a protein concentration of 0.96 μg/ml. The highest peroxidase enzyme activity was measured at 41 U/min at 80 % cut using a 250 μg protein concentration, while the highest polyphenol oxidase activity of 7.16 μg/ml was observed in the 40 % cut fraction. The synthesis of EZnO nanoparticles was confirmed through a visible color change and UV–visible spectroscopy with a distinctive absorbance peak at 307. 40 nm. X-ray diffraction (XRD) analysis at 2θ angle indicated the crystalline structure of the nanoparticles, with characteristic peaks corresponding to the planes of the ZnO crystal lattice. Fourier-transform infrared spectroscopy (FTIR) characterization revealed the embedding of the peroxidase enzyme within the nanoparticles, evidenced by absorbance peaks at 1708.62 cm−1 corresponding to amide I, 1604.48 cm−1 corresponding to amide II, and 1380 cm−1 corresponding to amide III. Similarly, the presence of polyphenol oxidase was indicated by absorbance peaks at 3185.83 cm−1 and 2981.41 cm−1. Transmission electron microscopy (TEM) analysis showed the nanoparticles exhibited polydispersity with a size range of 10–100 nm, with an agglomerated morphology, suggesting successful enzyme-nanoparticle hybridization. The histogram was constructed based on the counting of nanoparticles with size which showed highest number of nanoparticles were in the range of between 10 and 20 nm. The BET analysis showed a surface area of 225.1 m2/g. The EZnO nanoparticles demonstrated effective detoxification capabilities, achieving 86 % removal of synthetic phenol, 63 % removal of synthetic 2-chlorophenol, and 69 % removal of their mixture within 24 h. These findings highlights the potential of L. camara L. fruit extract and EZnO nanoparticles for application in the detoxification of hazardous pollutants, offering a sustainable and efficient approach to environmental remediation.

Influenza and non-influenza ARVI in children. A relationship between cytokine profile, parameters of the “lipid peroxidation – antioxidant defense system” as well as clinical and laboratory indicators

Influenza and acute respiratory viral infections (ARVI) impose a substantial damage to the population health in the Russian Federation due to their seasonal circulation and predominantly affect young children. A very few data on the cytokine profile, the nonspecific LPO — AOD system and their relationships with clinical characteristics in such diseases in preschool children are available. The aim of this study was to assess the cytokine profile, LPO — AOD system parameters and their relationship with the clinical and laboratory characteristics of diseases in preschool children with influenza and other ARVI. 86 preschool children (3–6 years old) were examined: with an established diagnosis of influenza (n = 31), non-influenza ARVI (n = 28), apparently healthy children (control group (n = 27). All pediatric samples were analyzed by enzyme-linked immunosorbent assay assessing blood serum concentrations of C-reactive protein and cytokines IL-1β, IL-4, IL-6, IL-8, TNFα, IFNα, IFNγ. Spectrophotometric, fluorometric and enzyme immunoassay methods to assess the state of the “LPO — AOD” system were used. In the group of children with influenza vs other ARVI, a higher incidence of intoxication syndrome was revealed. Cytokine profile in children from both clinical groups compared with control cohort was featured with higher indicators of both pro-inflammatory and anti-inflammatory origin. Children with non-influenza ARVI, had increased magnitude of LPO final products in the nonspecific LPO — AOD system along with lowered concentration of fat-soluble vitamins, general antioxidant activity, GSH level, and SOD activity. In the group with influenza, the level of primary and final lipid peroxidation products was increased, whereas that of for retinol,α-tocopherol, and total antioxidant activity was decreased paralleled with higher GSSG and SOD levels. Numerous correlations were noted in the group of children with ARVI: IL-1β/ketones, IL-6/ketones, IL-8/ketones, TNFα/ketones, IL-4/ketones, IFNg/shortness of breath, IFNα/cough, double bonds/fever, double bonds/AST, SO/intoxication, retinol/fever, GSSG/cough. The influenza group differed in the following relationships: IL-4/ketones, IL-4/fever, IFNα/ketones, CDs/AST. It can be concluded that in preschool children with ARVI and influenza, changes in the cytokine profile are accompanied by increased pro- and anti-inflammatory cytokine levels, increased intensity of lipid peroxidation reactions along with reduced magnitude of antioxidant factors. In the group with ARVI, there was a relationship between the final toxic products of lipid peroxidation — Schiff bases — and the intoxication index, as well as the presence of protective mechanisms in the form of connections between interferons and disease clinical manifestations. The group with influenza was distinguished by the presence of protective relations, which may have a beneficial effect in the context of developing pathological process. The data obtained will help expand the understanding of the pathogenetic mechanisms related to immune reactivity and nonspecific lipid peroxidation reactions in preschool patients and formulate appropriate measures for correction.

Effect of Blood Sampling Time After Colostrum Intake on the Concentration of Metabolites Indicative of the Passive Immunity Transfer in Newborn Dairy Calves

This study aimed to evaluate the impact of time after birth on the concentrations of metabolites relevant to monitoring passive immune transfer in calves. Additionally, we aimed to identify the best measurement devices based on accuracy, speed, cost-effectiveness, and availability on farms. Calves received 6 L of high-quality colostrum within 6 h. Blood samples were taken before (0 h) and 1, 2, 4, 6, 12, 24, 48, 72, 96, and 120 h after colostrum intake. The total serum protein (TSP) concentration increased during the first hours after colostrum intake, reaching stability between 24 and 48 h. The γ-glutamyl transferase and alkaline phosphatase concentrations increased after 4 h, followed by a significant decrease, and stabilized after 24 h. The albumin concentration decreased from 0 to 12 h then increased until 48 h and stabilized. Hematocrit showed higher values one hour after intake and remained constant after 24 h. Total serum protein stabilized at 24–48 h, which was the optimal window for passive transfer assessment. Digital or optical refractometers are recommended for TSP measurement, showing a strong correlation with enzyme methods, though initial calibration is necessary due to brand variations. These devices offer faster results compared to enzyme methods and direct IgG analysis.

The Ontogeny and Dietary Differences in Queen and Worker Castes of Honey Bee (Apis cerana cerana)

The honey bee Apis cerana cerana (A. c. cerana), a subspecies of Apis cerana, is endemic in China and possesses a valuable ecological niche. Understanding the ways to protect this honey bee’s populations is crucial, but this topic has been understudied. For the efficient utilization of beekeeping and pollination, there is a need to explore its biology and management practices. In light of this, the current study was carried out to investigate the ontogeny and dietary differences in the queen and worker castes of the A. c. cerana honey bee. This article presents, supplemented by reference images, a detailed description of the life history of A. c. cerana queens and workers. Additionally, this study investigated the nutritional differences between royal jelly (RJ) and worker jelly (WJ) at various larval ages. The contents of the moisture, crude protein, and amino acids in RJ and WJ were determined via freeze drying, Kjeldahl nitrogen determination, and ultra-high performance liquid chromatography. The results highlight significant variations in the moisture content, crude protein concentration, and amino acid concentration between RJ and WJ. The results offer theoretical support for ex situ artificial rearing practices of A. c. cerana.