Soluble Fraction Research Articles

Increased Homer Activity and NMJ Localization in the Vestibular Lesion het-/- Mouse soleus Muscle.

We investigated the shuttling of Homer protein isoforms identified in soluble (cytosolic) vs. insoluble (membrane-cytoskeletal) fraction and Homer protein-protein interaction/activation in the deep postural calf soleus (SOL) and non-postural gastrocnemius (GAS) muscles of het-/- mice, i.e., mice with an autosomal recessive variant responsible for a vestibular disorder, in order to further elucidate a) the underlying mechanisms of disrupted vestibular system-derived modulation on skeletal muscle, and b) molecular signaling at respective neuromuscular synapses. Heterozygote mice muscles served as the control (CTR). An increase in Homer cross-linking capacity was present in the SOL muscle of het-/- mice as a compensatory mechanism for the altered vestibule system function. Indeed, in both fractions, different Homer immunoreactive bands were detectable, as were Homer monomers (~43-48 kDa), Homer dimers (~100 kDa), and several other Homer multimer bands (>150 kDA). The het-/- GAS particulate fraction showed no Homer dimers vs. SOL. The het-/- SOL soluble fraction showed a twofold increase (+117%, p ≤ 0.0004) in Homer dimers and multimers. Homer monomers were completely absent from the SOL independent of the animals studied, suggesting muscle-specific changes in Homer monomer vs. dimer expression in the postural SOL vs. the non-postural GAS muscles. A morphological assessment showed an increase (+14%, p ≤ 0.0001) in slow/type-I myofiber cross-sectional area in the SOL of het-/- vs. CTR mice. Homer subcellular immuno-localization at the neuromuscular junction (NMJ) showed an altered expression in the SOL of het-/-mice, whereas only not-significant changes were found for all Homer isoforms, as judged by RT-qPCR analysis. Thus, muscle-specific changes, myofiber properties, and neuromuscular signaling mechanisms share causal relationships, as highlighted by the variable subcellular Homer isoform expression at the instable NMJs of vestibular lesioned het-/- mice.

Fabrication, optimization, and in vitro validation of penicillin-loaded hydrogels for controlled drug delivery

Bacterial infections present a major global challenge. Penicillin, a widely used antibiotic known for its effectiveness and safety, is frequently prescribed. However, its short half-life necessitates multiple high-dose daily administrations, leading to severe side-effects. Therefore, this study aims to address these issues by developing hydrogels which control the release of penicillin and alleviate its adverse effects. Various combinations of aspartic acid and acrylamide were crosslinked by N′, N′-methylene bisacrylamide through a free radical polymerization process to prepare aspartic acid/acrylamide (Asp/Am) hydrogels. The fabricated hydrogels underwent comprehensive characterization to assess physical properties and thermal stability. The soluble and insoluble fractions and porosity of the synthesized matrix were evaluated by sol-gel and porosity studies. Gel fraction was estimated at 88–96%, whereas sol fraction was found 12–4% and porosity found within the 63–78% range for fabricated hydrogel formulations. Maximum swelling and drug release were seen at pH 7.4, demonstrating a controlled drug release from hydrogel networks. The results showed that swelling, porosity, gel fraction, and drug release increased with higher concentrations of aspartic acid and acrylamide. However, integration of N′, N′-methylene bisacrylamide exhibited the opposite effect on swelling and porosity, while increasing gel fraction. All formulations followed the Korsymer–Peppas model of kinetics with ‘r’ values within the range of 0.9740–0.9980. Furthermore, the cytotoxicity study indicated an effective and safe use of hydrogel because the cell viability was higher than 70%. Therefore, these prepared hydrogels show promise candidates for controlled release of Penicillin and are anticipated to be valuable in clinical applications.

Comparison of acid and hydrothermal pretreatments of date waste for value creation

The production of date syrup yields a substantial amount of date press cake (DPC), fibrous and moisturising material with great potential for generating value through bioprocessing. However, the recalcitrant structure of DPC affects the yield of products in bioprocesses. To boost the accessibility of the structure as well as increase the soluble fraction of carbohydrates and facilitate further enzymatic hydrolysis, hydrothermal and dilute acid (0.5% (v/v) sulfuric acid) pretreatments as cost-effective and feasible methods were applied on DPC at relatively low temperatures (80, 100, 120 and 140 °C) and reaction times (60 and 90 min). The success in pretreatment was then evaluated by a post-enzymatic treatment using an enzyme cocktail of cellulases and hemicelluloses. Based on total accessible sugar with minimum produced inhibitors, an optimal operating condition was considered acid pretreatment at 120 °C for 90 min with a 55.02% increase in total sugar yield. To explore the potential use of pretreated DPC, an anaerobic digestion was conducted on untreated and acid-pretreated DPC at 120 °C for 90 min. The results showed that pretreatment increased the total bioproduct yield, including hydrogen, ethanol, and volatile fatty acid yields, by 59.75%. This demonstrates the significant impact of pretreatment on product yields in a bioprocess.

Kraft lignin-based polyurethanes: Bulk properties, stability and adhesion to native aluminum surfaces

Lignin as a byproduct from pulp and paper manufacturing has been extensively investigated as raw material for polymer developments. Although, fundamental topics related to the adhesion mechanisms of lignin-based reactive polyurethanes (LPU) still remain unclear. In this work, LPU thin films on native aluminum surface were prepared and characterized. Diluted in THF, employed as the solvent, reactive mixtures of 4,4′-methylene diphenyl isocyanate (4,4′-MDI) and the soluble fraction of three lignins (liquid hydroxypropylated Kraft lignin and two powder Kraft lignins with different pH) were used for LPU thin film deposition via spin coating on aluminum (PVD layer on silicon wafer). The resulting film thickness ranged from 8 nm to several micrometers. The chemical state of the three LPU compositions was assessed in bulk by infrared attenuated total reflectance (IR-ATR) and in the thin films by infrared external reflection absorption spectroscopy (IR-ERAS). Binding energies of 4,4′-methylene diphenyl diisocyanate with aliphatic and aromatic hydroxyl groups were estimated using Density Functional Theory (DFT) simulations. Thus, besides the elucidation of the bulk chemical state of LPUs, the relation to adhesion and stability of LPUs to a native aluminum surface was evaluated. In addition, film topography and homogeneity were monitored by SFM. All lignin types form uniform and homogeneous films. Results revealed a higher consumption of isocyanate groups (NCO) in the formulation with the alkaline Kraft lignin than with the acid one, despite its lower hydroxyl content. A gradient of unreacted NCO was observed in LPU thin films obtained with the powder Kraft lignin types. Residual NCO is also found in thicker films, while the thinnest films did not contain unreacted NCO anymore, indicating the activating effect of the native aluminum surface on LPU formation.

Abstract We054: Mitochondria-containing Extracellular Vesicles Mediate Heart Failure Sterile Inflammation

Introduction: Sterile inflammation has long been postulated to exacerbate heart failure (HF); however, a lack of viable biological targets and thorough understanding of the underlying mechanism posed formidable challenges in the development of novel therapeutic interventions. Hypothesis: Given proinflammatory mitochondrial DNA (mtDNA) are known to be elevated in HF plasma; and that cells release mitochondrial components in extracellular vesicles, we hypothesize that proinflammatory circulating Mitochondria-containing Extracellular Vesicle (MitoEV) is a mediator of HF sterile inflammation. Methods/Results: Using plasma samples of 12 Stage-D HFrEF and 12 healthy participants, plasma microparticles (MP) are co-stained with mitochondrial- and cytoplasmic-specific markers then quantified by volume-metric flow analysis. We found that there are significantly more mitochondrial-cytoplasmic double-positive MP (designated MitoEV, accounts for >70% of total MP) in HF relative to control. Next, using size-exclusion chromatography, we found MitoEV fractions are significantly more potent than soluble protein fractions to induce cytokine production in macrophage, implicating MitoEV as the source of HF plasma immunogenicity. Surface marker analysis by flow suggests that CD14+ monocytes is a major source of HF plasma MitoEV. Using THP-1 monocytes, we found that TLR9 activation by mtDNA results in injured mitochondria to be released as MitoEV (rather than removed by mitophagy), a pathway negatively regulated by beclin-1. Finally, boosting NAD by nicotinamide riboside preserves mitochondrial dysfunction and significantly reduces MitoEV release in primary monocytes. Conclusion: Our findings implicate circulating MitoEV released by monocytes upon mtDNA activation as a pivotal mediator of sterile inflammation in HF, demonstrating a promising avenue for potential therapeutic targeting. The study also provides a potential cellular mechanism to the anti-inflammatory effect of boosting NAD. Model: In HF circulation, mtDNA activates TLR9 of monocytes, injuring mitochondria, which are released as MitoEV (containing mtDNA) to amplify and propagate inflammation in an autocrine manner. Becn1, a key mediator of mitophagy, negatively regulates MitoEV release. Activated monocytes also release cytokines to further injure the heart, resulting in a vicious cycle. Boosting NAD protects against mtDNA-induced mitochondrial dysfunction, attenuates the MitoEV release, and ameliorates inflammation.

Preferential Solvation of Cyclosporin in Aqueous Mixtures of Some Polymeric Cosolvents

Background: Cyclosporin is a cyclic peptide-drug used as immunosuppressant for the prophylaxisof transplant rejection whose physicochemical properties in mixed aqueous solvent systems isstill not well understood. The preferential solvation parameters of cyclosporin in aqueous binarymixtures of diethylene glycol monoethyl ether (DEGME), polyethylene glycol 200 (PEG 200) andpolyethylene glycol 400 (PEG 400) were computed. Methods: Reported mole fraction solubilities of cyclosporin in DEGME-aqueous mixtures,PEG 200-aqueous mixtures, and PEG 400-aqueous mixtures were processed by following theinverse Kirkwood-Buff integrals (IKBI) method as suggested by Marcus and Ben-Naim using somethermodynamic parameters reported in the literature for these aqueous-polymeric mixtures at298.15 K. Results: It is observed that cyclosporin is sensitive to preferential solvation effects in theseaqueous-polymeric binary solvent systems. The preferential solvation parameter by DEGME (δx1,3)is negative in water-rich mixtures but positive in mixtures of 0.12<x1<1.00. It is conjecturablethat hydrophobic hydration around the non-polar methyl and methylene groups of this drug thatcould be present in water-rich mixtures can significantly impact the drug solvation. Otherwise,in mixtures of 0.12<x1<1.00 in DEGME-aqueous mixtures, as well as in almost all the mixtureswith PEGs, the preferential solvation by polymeric cosolvents could be due to the acidic behaviorof cyclosporin in front of ether and hydroxyl oxygen atoms of these polymeric cosolvents. Conclusion: Cyclosporin is preferentially solvated by the polymeric solvents in almost all thestudied mixtures of these aqueous-polymeric binary solvent systems.

Iron-binding biomolecules in the soluble hepatic fraction of the northern pike (Esox lucius): two-dimensional chromatographic separation with mass spectrometry detection.

Iron plays vital roles in important biological processes in fish, but can be toxic in high concentrations. The information on metalloproteins that participate in maintenance of Fe homeostasis in an esocid fish, the northern pike, as an important freshwater bioindicator species, are rather scarce. The aim of this study was to identify main cytosolic constituents that sequester Fe in the northern pike liver. The method applied consisted of two-dimensional HPLC separation of Fe-binding biomolecules, based on anion-exchange followed by size-exclusion fractionation. Apparent molecular masses of two main Fe-metalloproteins isolated by this procedure were ~360 kDa and ~50 kDa, with the former having more acidic pI, and indicated presence of ferritin and hemoglobin, respectively. MALDI-TOF-MS provided confirmation of ferritin subunit with a m/z peak at 20.65 kDa, and hemoglobin with spectra containing main m/z peak at 16.1 kDa, and smaller peaks at 32.1, 48.2, and 7.95 kDa (single-charged Hb-monomer, dimer, and trimer, and double-charged monomer, respectively). LC-MS/MS with subsequent MASCOT database search confirmed the presence of Hb-β subunits and pointed to close relation between esocid and salmonid fishes. Further efforts should be directed towards optimization of the conditions for metalloprotein analysis by mass spectrometry, to extend the knowledge on intracellular metal-handling mechanisms.

Deciphering the uranium isotopic signature of coastal water and sediments from Tokyo Bay using a multi-collector inductively coupled plasma mass spectrometer

We have measured 236U/238U and 235U/238U values in JMS-1 (geochemical reference material of Tokyo Bay sediments) and coastal seawater using a multi-collector inductively coupled plasma mass spectrometer (MC-ICP-MS) equipped with both the retarding potential quadrupole lens and desolvating nebuliser system. For the accurate measurement of 236U/238U values (e.g., in the range of 10−9), mass spectrometric interferences on 236U isotope from both the peak tailing of 238U and polyatomic ion of 235UH were carefully corrected. With the sequential extraction experiments for JMS-1, whose U isotopic signatures were characteristic of isotopically-depleted U with industrial uses, authigenic U was extracted into a soluble fraction, and lithogenic U and anthropogenic U were gathered in an insoluble fraction. The anthropogenic U is likely to have been provided in insoluble forms and have deposited on the bay floor. Absence in differences of 236U/238U and 235U/238U values for seawater observed between inside and outside Tokyo Bay implies the negligibly small contribution of the anthropogenic U to U in the seawater. The data obtained here demonstrate the effective reduction of the interferences on 236U and the versatility of the isotopic signatures of U as an effective tracer for environmental circulation of U in nature.

The Effect of Covering Corn Silage with Tomato or Apple Pomace on Fermentation Parameters and Feed Quality

The current study assessed the effects of covering corn silage with tomato or apple pomace on fermentability and feed quality. The in vitro gas production test was performed using graded 100 mL syringes. Incubation times were 3, 6, 12, 24, 48, 72, and 96 h. In vitro gas generation characteristics were significantly altered by TP (tomato pomace) and AP (apple pomace), both alone and in conjunction with PE (polyethylene) films, regardless of their presence. As a result of the effects found on NH3-N concentration, aerobic stability, and yeast activity, TP and AP have the potential to become an eco-friendly alternative to PE films. The gas production from the immediately soluble fraction (a) of corn silage was only affected when the corn silage was covered with a combination of AP and PE compared to the CPE group (p < 0.001). The largest cluster includes correlations of the DOM-TDDM (r = 0.90), DOM-AA (r = 0.88), and Ash-TDDM (r = 0.86) correlations. The most substantial negative correlations were identified between DM-CO2 (r = −82), DM-Yeast (r = −0.79), and CF-DOM (r = −0.79). Nonetheless, the use of pomace as a silage cover presents an inexpensive alternative to plastic films for silage that does not have the environmental problems associated with persistent micro- and nanoplastics.

In Vitro Phytochemical Analysis of Antioxidant and Antimicrobial Properties of Tephrosia Purpurea

Tephrosia purpurea is a flowering plant that has a pantropical distribution. It is throughout found in india and srilanka A decoction of the roots is given in dyspepsia, diarrhea, rheumatism, asthma and urinary disorders. Two native plants, Tephrosia purpurea (Linn.) Pers. (Fabaceae) and Mimusops elengi (Linn.) (Sapotaceae) were screened for their antimicrobial activity. Preliminary testing of antimicrobial activity of T. purpurea against 3 standard cultures In addition, ethonolic extracts were prepared from the bark of M. elengi and tested for its antimicrobial activity against the above bacterial isolates. Present work was undertaken to study antioxidant potential of the plant. It can be concluded that the aqueous extract of T. purpurea L. root can be used as antioxidant and it can be recommended for the treatment of various disease. The study concluded that the ethanolic extract of T. purpurea exhibits antioxidant activity in vivo and the ethyl acetate soluble fraction has improved antioxidant potential than the extract.

Solubility of curcumin in subcritical water: Experimental data, modeling, and thermodynamic parameters

As an anticancer drug, the mole fraction solubility of curcumin in subcritical water (SBCW) was determined by a static method. Experiments were performed at temperatures between 298.15–423.15 K and a constant pressure of 20 bar. With increasing temperature, the mole fraction of solubility showed a significant increase in the range of 0.03 × 10-6< x 2< 13.22 × 10-6. For the correlation of experimental solubility data, linear, modified Apelblat, NRTL, UNIQUAC, and Wilson thermodynamic models were selected. Among the models, NRTL and modified Apelblat models had a better correlation with the experimental data with MPD% 6.94, 4.68, and RMSD×105 0.07, 0.04, respectively. Also, the thermodynamic functions such as Gibbs free energy, enthalpy, and entropy were calculated for the solubility process of curcumin in SBCW.

Tau protein profiling in tauopathies: a human brain study

Abnormal accumulation of misfolded and hyperphosphorylated tau protein in brain is the defining feature of several neurodegenerative diseases called tauopathies, including Alzheimer’s disease (AD). In AD, this pathological change is reflected by highly specific cerebrospinal fluid (CSF) tau biomarkers, including both phosphorylated and non-phosphorylated variants. Interestingly, despite tau pathology being at the core of all tauopathies, CSF tau biomarkers remain unchanged in certain tauopathies, e.g., progressive supranuclear palsy (PSP), Pick’s disease (PiD), and corticobasal neurodegeneration (CBD). To better understand commonalities and differences between tauopathies, we report a multiplex assay combining immunoprecipitation and high-resolution mass spectrometry capable of detecting and quantifying peptides from different tau protein isoforms as well as non-phosphorylated and phosphorylated peptides, including those carrying multiple phosphorylations. We investigated the tau proteoforms in soluble and insoluble fractions of brain tissue from subjects with autopsy-confirmed tauopathies, including sporadic AD (n = 10), PSP (n = 11), PiD (n = 10), and CBD (n = 10), and controls (n = 10). Our results demonstrate that non-phosphorylated tau profiles differ across tauopathies, generally showing high abundance of microtubule-binding region (MTBR)-containing peptides in insoluble protein fractions compared with controls; the AD group showed 12–72 times higher levels of MTBR-containing aggregates. Quantification of tau isoforms showed the 3R being more abundant in PiD and the 4R isoform being more abundant in CBD and PSP in the insoluble fraction. Twenty-three different phosphorylated peptides were quantified. Most phosphorylated peptides were measurable in all investigated tauopathies. All phosphorylated peptides were significantly increased in AD insoluble fraction. However, doubly and triply phosphorylated peptides were significantly increased in AD even in the soluble fraction. Results were replicated using a validation cohort comprising AD (n = 10), CBD (n = 10), and controls (n = 10). Our study demonstrates that abnormal levels of phosphorylation and aggregation do indeed occur in non-AD tauopathies, however, both appear pronouncedly increased in AD, becoming a distinctive characteristic of AD pathology.

Physical and Functional Properties of Powders Obtained during Spray Drying of Cyani flos Extracts.

Edible flowers are a potential source of bioactive ingredients and are also an area of scientific research. Particularly noteworthy are Cyani flos, which have a wide range of uses in herbal medicine. The below study aimed to investigate the influence of selected soluble fiber fractions on the selected properties of physical and biochemical powders obtained during spray drying a water extract of Cyani flos. The drying efficiency for the obtained powders was over 60%. The obtained powders were characterized by low moisture content (≤4.99%) and water activity (≤0.22). The increase in the addition of pectin by the amount of 2-8% in the wall material resulted in a decrease in hygroscopicity, water solubility, and protection of flavonoids and anthocyanins both before and after digestion in the tested powders in comparison to the sample with only inulin as a carrier. Additionally, it was noted that all samples were characterized by high bioaccessibility when determining antioxidant properties and xanthine oxidase inhibition.

Production of recombinant human epidermal growth factor fused with HaloTag protein and characterisation of its biological functions.

Epidermal growth factor (EGF) protein is a crucial biomolecule involved in regulating cell growth, proliferation, migration and differentiation, which is used in various therapeutic applications, such as wound healing and tissue regeneration. The production of recombinant EGF is essential for studying its biological function and for its clinical translation. However, EGF protein expressed in prokaryotic cells often occurs in inclusion bodies, and co-expression with soluble tag protein is an effective method to prepare recombinant EGF. In this study, we expressed recombinant human EGF (rhEGF) fused to a HaloTag (Halo-rhEGF) and a large portion of Halo-rhEGF was found in the soluble fraction. Cell growth assay showed that the purified Halo-rhEGF protein could promote the proliferation of fibroblasts (NIH 3T3) and epithelial cells (HaCaT), and significantly increased their viability. Phosphorylation of the intracellular signaling proteins, ERK1/2 and c-Jun, was stimulated by treatment with Halo-rhEGF and the expression levels of proteins regulating cell proliferation were significantly increased. RNA sequencing analysis revealed that rhEGF could increase the transcription of genes enriched in ribosome generation and cell proliferation. Moreover, Halo-rhEGF can be labelled by HaloTag ligand for fluorescence imaging and can be slowly released in tissue repair by binding to anion biomaterials. In conclusion, HaloTag is an efficient fusion tag for rhEGF protein expression, purification and controlled release, and Halo-rhEGF can promote the proliferation and viability of epithelial and fibroblast cells.

Effect of different pectin sources and structures on the stability of cyanidin-3-O-glucoside (C3G)

The physicochemical properties of cyanidin-3-O-glucoside (C3G) were investigated after binding with pectin from different sources (apple and peach) and fractions (water-soluble fraction (WSF), chelator soluble fraction (CSF) and sodium carbonate soluble fraction (NSF)). Apple WSF and peach CSF were high esterification degree pectin (DE) (63.23% and 55.66%). CSF of both AP and PP was abundant in HG and exhibited a much more linear structure than WSF and NSF. The maximum binding percentage was obtained at pH 2.0 for apple pectin (AP) (46%–50%) and at pH 3.5 for peach pectin (PP) (50%–54%). Binding with both pectins increased the color stability of C3G at pH 2.0. High DE pectin increased C3G light stability by reducing the degradation rate (0.03 h−1) compared to C3G alone (0.05 h−1). All the pectins reduced the thermal degradation rate of C3G, thus exhibiting a protective effect. A negative correlation was found between the relative RG I area and the particle size of the pectin-C3G complexes. Pectin played a leading role in the ζ-potential change.

Investigating the synergistic potential of Si and biochar to immobilize Ni in a Ni-contaminated calcareous soil after Zea mays L. cultivation

Abstract. In Iran, a significant percentage of agricultural soils are contaminated with a range of potentially toxic elements (PTEs), including Ni, which need to be remediated to prevent their entry into the food chain. Silicon (Si) is a beneficial plant element that has been shown to mitigate the effects of PTEs on crops. Biochar is a soil amendment that sequesters soil carbon and that can immobilize PTEs and enhance crop growth in soils. No previous studies have examined the potentially synergistic effect of Si and biochar on the Ni concentration in soil chemical fractions and the immobilization thereof. Therefore, the aim of this study was to examine the interactive effects of Si and biochar with respect to reducing Ni bioavailability and its corresponding uptake in corn (Zea Mays) in a calcareous soil. A 90 d factorial greenhouse study with corn was conducted. Si application levels were 0 (S0), 250 (S1), and 500 (S2) mg Si kg−1 soil, and biochar treatments (3wt %) including rice husk (RH) and sheep manure (SM) biochars produced at 300 and 500 °C (SM300, SM500, RH300, and RH500) were utilized. At harvest, the Ni concentration in corn shoots, the Ni content in soil chemical fractions, and the release kinetics of DPTA (diethylenetriaminepentaacetic acid)-extractable Ni were determined. Simultaneous utilization of Si and SM biochars led to a synergistic reduction (15 %–36 %) in the Ni content in the soluble and exchangeable fractions compared with the application of Si (5 %–9 %) and SM (5 %–7 %) biochars separately. The application of Si and biochars also decreased the DPTA-extractable Ni and Ni content in corn shoots (by up to 57 %), with the combined application of SM500 + S2 being the most effective. These effects were attributed to the transfer of Ni in soil from more bioavailable fractions to more stable iron-oxide-bound fractions, related to soil pH increase. SM500 was likely the most effective biochar due to its higher alkalinity and lower acidic functional group content which enhanced Ni sorption reactions with Si. The study demonstrates the synergistic potential of Si and SM biochar for immobilizing Ni in contaminated calcareous soils.

High hydrostatic pressure for decontamination of soluble insect proteins prevents protein denaturation better than blanching

Insect production currently involves thermal processes to ensure microbial safety and inactivate endogenous enzymes, but these processes negatively affect the protein's technological properties. Therefore, this study compared high hydrostatic pressure (HHP) at 200/400/600 MPa for 10 min to blanching at 90 °C for 10 min, on soluble protein fractions (pH 3) from house crickets (Acheta domesticus) and lesser mealworms (Alphitobius diaperinus). HHP significantly reduced the microbial load, especially at 400 and 600 MPa. For aerobic counts of mealworms, blanching (from ∼6 to ∼3 log CFU/mL) was more effective than HHP (∼4 log CFU/mL). HHP preserved the secondary protein structure better than blanching, but was less effective in protease inactivation, though only evident in mealworms. HHP did not affect foaming and emulsification properties but improved protein solubility after pH adjustment to 7 compared to untreated fractions by 14–22%. We thus demonstrate that HHP can be an effective alternative to conventional blanching treatments. Industrial relevanceThe emerging edible insect industry is currently struggling with several challenges. Conventional thermal processes that are often applied can negatively affect the techno-functional protein properties. We demonstrated that high hydrostatic pressure (HHP) can be an effective milder alternative to thermal processes. The evaluated blanching condition did not always negatively affect the functional protein properties, and blanching was more effective in protease inactivation compared to HHP for lesser mealworms. HHP can be of industrial relevance for inactivating house cricket fractions with high protein solubility intended for emulsion applications. Blanching could be better suited to provide proteolytically stable fractions.

Nutritional and sensory parameters of amazake from the recycling of stale bread

Stale bread is a waste product with a potential to be recycled. One way to manage this waste material is to process it by fermentation for the purpose of food production. This paper proposes the use of stale wheat and rye bread as ingredients in amazake, a liquid dessert traditionally obtained from rice by fermentation with the koji mould Aspergillus oryzae, followed by liquefaction by the action of fungal enzymes. The stale bread was introduced instead of rice at both the koji stage (wheat bread) and the liquefaction stage (wheat and rye bread). The resulting products had an extended volatile compound profile, from 5 to 15 compounds identified, and modified sensory parameters, compared to the traditional version. Amazake containing bread had an increased protein content, from 1.10 to 6.4 g/100 g, and were more abundant in dietary fibre (up to a maximum of 1.8 g/100 g), additionally enriched with a soluble fraction. The proposed procedure of obtaining of new formula amazake can be directly applied in households to reduce the amount of discarded bread. Due to its simplicity, it also has the potential for further modification in terms of production scale and product parameters.

Seasonal Dynamic of NO3− and K+ in a Citrus Crop Irrigated by Different Water Qualities

This study evaluated the effect of (i) irrigation water source: transfer (TW) and reclaimed water (RW), and (ii) crop phenological stage: winter rest (WR), flowering-sprouting (FS), and fruit growth (FG), on NO3− and K+ dynamics in soil and leaf of a citrus crop. The experiment was carried out during the 2018 and 2019 growing seasons on adult ‘Star Ruby’ grapefruit trees (Citrus paradisi Macf.). The concentration of both nutrients was periodically measured in soil and leaf samples and continuously monitored in the soil soluble fraction with nutrient sensors. Moreover, soil NO3− leaching was indirectly estimated by the periodic measurement of the leaf enrichment in 15N isotope (15Nleaf). The two water sources showed a different nutrient loading. Thus, NO3− and K+, were approximately 5 and 7 times higher, respectively, in the RW. Furthermore, the average contents of NO3− and K+ in the soil samples from the RW treatment were 10.1 and 19.7%, respectively, higher than in TW, with the highest soil NO3− leaching observed in RW treatment. In line with this, the mean contents of NO3− and K+ in the leaves from the RW treatment were 106.9 and 30.4% higher than the TW ones. As for the different phenological stages, in the FG stage, the lowest concentrations of NO3− in the soil samples and the highest in the leaf tissue were observed after a high soil leaching event. In this study the nutrient sensors measurements varied according to the dynamic of NO3− and K+ in the soil samples. The use of RW promoted an accumulation of NO3− and K+ in the soil and leaves of grapefruit trees, but also enhanced soil NO3− leaching, indicating that the proper management of this water source is necessary to avoid soil contamination. The mobilization of NO3− and K+ from soil to leaf was the highest in the FG stage, to ensure fruit development and vegetative growth.

Systemic isotretinoin in modern clinical guidelines on acne management

Objective — to analyze the medical and technological documents developed on the basis of evidence­based medicine (guidelines, recommendations, consensuses) regarding the treatment of patients with acne of various degrees of severity with systemic retinoids (systemic isotretinoin preparations) dating from 2013 to 2023, as also to focus attention on the pharmacokinetic features of the original systemic isotretinoin according to innovative Lidose technology. Materials and methods. An analysis was conducted of clinical European guidelines (French guideline and those of the European Dermatology Forum), guidelines of the American Academy of Dermatology, the National Institute for Health Protection and Improvement of Medical Care (NICE) of Great Britain in conjunction with the British Dermatology Association and the Canadian Dermatology Association, recommendations of the American Academy of Pediatrics and the French Society of Dermatology, the International Consensus of the Global Alliance for the Improvement of Acne Treatment Outcomes, and consensus­based practical and daily guideline for the management of acne patients. The key words for the search were «acne», «clinical guideline», «consensus», «systemic treatment», «isotretinoin», «isotretinoin­ Lidose». Results and discussion. Clinical guidelines (2013, 2015, 2016) for the management of acne recommend isotretinoin at a dose of 0.5—1 mg/kg/day for 16—24 weeks to a total (cumulative) dose of 120—150 mg/kg for severe acne (nodular/conglobate acne). Low-­dose regimens of isotretinoin (&lt; 0.5 mg/kg/day) have been used with varying degrees of success in the treatment of severe papulopustular/moderate nodular acnе. The International Consensus of the Global Alliance to Improve Acne Treatment Outcomes (2017) supports standard isotretinoin dosing for severe acne until the cumulative dose is reached. At the same time, for patients that have severe papulopustular acne, poor tolerance, risk of serious side effects, or rash regression before the cumulative dose is achieved it is recommended to continue taking the drug until the skin is completely cleansed of acne­elements and for additional 1—2 months. Such provisions are set out in the guidelines developed jointly with the British Association of Dermatologists (2021). According to the American Academy of Dermatology (2023) guidelines, standard isotretinoin or isotretinoin ­Lidose® is recommended for patients for whom isotretinoin is indicated, which is supported by high­quality evidence. Due to the Lidose technology, the soluble fraction of isotretinoin increases, while the insoluble fraction responsible for side effects, on the contrary, decreases. The bioavailability of the drug increases, which allows lowering the daily and course doses of the drug. Conclusions. Systemic isotretinoin has a convincing evidence base as highly effective treatment for severe and moderate forms of acne, providing a sustained therapeutic effect at standard or low doses. In current US guidelines, isotretinoin and isotretinoin­ Lidose® are recommended for the treatment of severe acne, as well as acne that causes psychosocial burden or scarring, or acne that does not respond to treatment with oral or topical therapy. The use of isotretinoin ­Lidose® due to extrabioavailability, high solubility, independence from food intake allows to obtain better clinical results, has a favorable safety profile, characterized by a minimal frequency of adverse reactions.