Protein Retention Research Articles

Combining single-molecule and expansion microscopy in fission yeast to visualize protein structures at the nanostructural level.

In this work, we have developed an expansion microscopy (ExM) protocol that combines ExM with photoactivated localization microscopy (ExPALM) for yeast cell imaging, and report a robust protocol for single-molecule and expansion microscopy of fission yeast, abbreviated as SExY. Our optimized SExY protocol retains about 50% of the fluorescent protein signal, doubling the amount obtained compared to the original protein retention ExM (proExM) protocol. It allows for a fivefold, highly isotropic expansion of fission yeast cells, which we carefully controlled while optimizing protein yield. We demonstrate the SExY method on several exemplary molecular targets and explicitly introduce low-abundant protein targets (e.g. nuclear proteins such as cbp1 and mis16, and the centromere-specific histone protein cnp1). The SExY protocol optimizations increasing protein yield could be beneficial for many studies, when targeting low abundance proteins, or for studies that rely on genetic labelling for various reasons (e.g. for proteins that cannot be easily targeted by extrinsic staining or in case artefacts introduced by unspecific staining interfere with data quality).

Bovine Serum Albumin Rejection by an Open Ultrafiltration Membrane: Characterization and Modeling.

The classic application of ultrafiltration (UF) is for the complete retention of proteins, and in that situation, the transport behavior is well established. More open membranes with fractional retention are used when separating different proteins. However, protein transport has not been well documented yet in the literature. The bovine serum albumin (∼69 kDa) observed rejection ranges from 0.65 to 1 using a 300 kDa molecular weight cut-off membrane at different pH, ionic strength, and pressure. We demonstrated that, especially with open UF, the transport of proteins through the membrane is dominated by advection, with insignificant diffusion effects (p value > 0.05). We showed that with open UF, retention is not only caused by size exclusion but also to a large extent by electrostatic interactions and oligomerization of the proteins. Mass transfer in the polarization layer was relatively independent of the pH and ionic strength. It was underestimated by common Sherwood relations due to a relatively large contribution of the reduction in the flow turbulence near the membrane by the removal of fluid through the membrane. We propose a model that allows relatively quick characterization of the rejection of proteins without prior knowledge of the pore sizes and charges based on just a limited set of experiments. Therefore, protein rejection with the open UF system can be targeted by tuning the processing conditions, which might be useful for designing protein fractionation processes.

A familial missense variant in the Alzheimer’s disease gene SORL1 impairs its maturation and endosomal sorting

The SORL1 gene has recently emerged as a strong Alzheimer’s Disease (AD) risk gene. Over 500 different variants have been identified in the gene and the contribution of individual variants to AD development and progression is still largely unknown. Here, we describe a family consisting of 2 parents and 5 offspring. Both parents were affected with dementia and one had confirmed AD pathology with an age of onset > 75 years. All offspring were affected with AD with ages at onset ranging from 53 years to 74 years. DNA was available from the parent with confirmed AD and 5 offspring. We identified a coding variant, p.(Arg953Cys), in SORL1 in 5 of 6 individuals affected by AD. Notably, variant carriers had severe AD pathology, and the SORL1 variant segregated with TDP-43 pathology (LATE-NC). We further characterized this variant and show that this Arginine substitution occurs at a critical position in the YWTD-domain of the SORL1 translation product, SORL1. Functional studies further show that the p.R953C variant leads to retention of the SORL1 protein in the endoplasmic reticulum which leads to decreased maturation and shedding of the receptor and prevents its normal endosomal trafficking. Together, our analysis suggests that p.R953C is a pathogenic variant of SORL1 and sheds light on mechanisms of how missense SORL1 variants may lead to AD.

Overexpression of ameloblastin in secretory ameloblasts results in demarcated, hypomineralized opacities in enamel.

Introduction: Developmental defects of the enamel manifest before tooth eruption and include amelogenesis imperfecta, a rare disease of underlying gene mutations, and molar-incisor hypomineralization (MIH), a prevalent disease in children originating from environmental and epigenetic factors. MIH enamel presents as the abnormal enamel marked by loss of translucency, demarcation between the healthy and affected enamel, and reduced mineral content. The pathophysiology of opaque, demarcated enamel lesions is not understood; however, the retention of enamel proteins in the matrix has been suggested. Ameloblastin (Ambn) is an enamel protein of the secreted calcium-binding phosphoproteins (SCPPs) critical for enamel formation. When the Ambn gene is mutated or deleted, teeth are affected by hypoplastic amelogenesis imperfecta. Methods: In this study, enamel formation in mice was analyzed when transgenic Ambn was overexpressed from the amelogenin promoter encoding full-length Ambn. Ambn was under- and overexpressed at six increasing concentrations in separate mouse lines. Results: Mice overexpressing Ambn displayed opaque enamel at low concentrations and demarcated lesions at high concentrations. The severity of enamel lesions increased starting from the inner enamel close to the dentino-enamel junction (DEJ) to span the entire width of the enamel layer in demarcated areas. Associated with the opaque enamel were 17-kDa Ambn cleavage products, a prolonged secretory stage, and a thin basement membrane in the maturation stage. Ambn accumulations found in the innermost enamel close to the DEJ and the mineralization front correlated with reduced mineral content. Demarcated enamel lesions were associated with Ambn species of 17kDa and higher, prolonged secretory and transition stages, a thin basement membrane, and shortened maturation stages. Hypomineralized opacities were delineated against the surrounding mineralized enamel and adjacent to ameloblasts detached from the enamel surface. Inefficient Ambn cleavage, loss of contact between ameloblasts, and the altered basement membrane curtailed the endocytic activity; thus, enamel proteins remained unresorbed in the matrix. Ameloblasts have the ability to distinguish between Ambn concentration and Ambn cleavage products through finely tuned feedback mechanisms. The under- or overexpression of Ambn in murine secretory ameloblasts results in either hypoplastic amelogenesis imperfecta or hypomineralization with opaque or sharply demarcated boundaries of lesions, similar to MIH.

Optimization of Process Parameters for the Production of Soy Protein by Ultrafiltration Using ANN

The growing popularity of soy proteins among vegans and vegetarians, owing to their high protein content and widespread availability, has led to scientific studies on its various extraction methods mainly on ultrafiltration. This research employed artificial neural network (ANN) and Box-Behnken design (BBD) methodologies to predict the process parameters of ultrafiltration for the preparation of soy protein. Using BBD, the optimum process parameters of ultrafiltration were identified via the desirability function approach. The optimized permeate flux was 11.13 litres per hour (LPH) and 85.52% protein content in retentate. The identified ideal process parameters for ultrafiltration to achieve maximal protein retention encompassed a 10 kDa membrane module, a transmembrane pressure of 117 kPa (17 PSI), a volume concentration ratio of 3.5, diafiltration set at 1, and a flow rate of 65% of the pump capacity, exhibiting an absolute percent error value of 2.81. Employing these refined process parameters, the predicted value for protein retentate stood at 80.49%. The predictive accuracy of the model achieved an impressive 99.61% for protein retention. The ANN model effectively predicted the optimal ultrafiltration conditions, resulting in maximal protein retention and a protein content accuracy of 96.41% and 99.61%, respectively.

Effects of Enzyme Treatment and Carrier Agents on Chemical and Physical Properties of Almond Protein-Based Product

There has been an emerging trend towards the research of plant-based proteins over the past few years; however, there has been limited conductive observation of plant-based proteins from almond by-products, as well as the potential of using Flavourzyme and different carrier agents to obtain an instant protein-based powder. The purpose of this study was to investigate the effects of Flavourzyme at various concentrations (0, 0.5, 1, 1.5, 2, and 2.5%) and incubation times (30, 60, 90, 120, and 180 min), as well as carrier agents at different ratios with a 20% total concentration on the physicochemical properties of almond protein-based products using freeze-drying (FD). The results showed a higher protein content (p-value ≤ 0.05) using 1.5% Flavourzyme for 120 min compared to the other levels. There was no significant difference (p-value > 0.05) in the protein retention rate after freeze-drying of the samples. However, the addition of Maltodextrin (MA), Gum Arabic (GA), and Inulin (IN) encapsulants improved the physical and functional characteristics of freeze-dried almond protein-based powder (FDAP). However, the solubility of the powder is moderate. Water-holding-capacity (WHC) and Oil- Holding Capacity (OHC) are inversely proportional, where GA-coated powder is the most hydrophobic (17.72 ± 0.87 mL oil/g) and IN-encapsulated powder is the most hydrophilic (15.00 ± 0.87 mL water/g). In conclusion, IN could be a potential encapsulant for almond protein-based powder, because powder produced from IN or MA: IN is acceptable in terms of physical parameters, and IN can also enhance fiber content in the final product.

Cancer Cell Targeting Via Selective Transferrin Receptor Labeling Using Protein-Derived Carbon Dots.

Carbon dot (CD) nanoparticles offer tremendous advantages as fluorescent probes in bioimaging and biosensing; however, they lack specific affinity for biomolecules, limiting their practical applications in selective targeting. Nanoparticles with intrinsic affinity for a target have applications in imaging, cytometry, therapeutics, etc. Toward that end, we report the transferrin receptor (CD71) targeting CDs, synthesized for the first time. The formation of these particles is truly groundbreaking, as direct tuning of nanoparticle affinity was achieved by simple and careful precursor selection of a protein, which has the targeting characteristic of interest. We hypothesized that the retention of the original protein's peptides on the nanoparticle surface provides the CDs with some of the function of the precursor protein, enabling selective binding to the protein's receptor. This was confirmed with FTIR (Fourier transform infrared) data and subsequent affinity-based cell assays. These transferrin (Tf)-derived CDs have been shown to possess an affinity for CD71, a cancer biomarker that is ubiquitously expressed in nearly every cancer cell line due to its central role mediating the uptake of cellular iron. The CDs were tested using the human leukemia cell line HL60 and demonstrated the selective targeting of CD71 and specific triggering of transferrin-mediated endocytosis via clathrin-coated pits. The particle characterization results reflect a carbon-based nanoparticle with bright violet fluorescence and 7.9% quantum yield in aqueous solution. These unpresented CDs proved to retain the functional properties of the precursor protein. Indicating that this process can be repeated for other disease biomarkers for applications ranging from biosensing and diagnostic bioimaging to targeted therapeutics.

Effect of different feeding strategies and dietary fiber levels on energy and protein retention in gestating sows.

The aim of the study was to investigate whether increased inclusion of sugar beet pulp (SBP) alters retention of fat, protein, and energy when backfat (BF) is restored in early- and mid-gestation. In total, 46 sows were fed one of four dietary treatments with increasing inclusion of SBP providing dietary fiber (DF) levels of 119, 152, 185, and 217g/kg; sows were assigned to one of three feeding strategies (FS; high, medium, and low) depending on BF thickness at mating and again at day 30 for the following month. On days 0, 30, 60, and 108, body weight (BW) and BF thickness were measured and body pools of protein and fat were estimated using the deuterium oxidetechnique. On days 30 and 60, urine, feces, and blood samples were collected to quantify metabolites, energy, and nitrogen (N) balances. On days 15 and 45, heart rate was recorded to estimate heat energy. At farrowing, total born and weight of the litter were recorded. In early gestation, BW gain (P < 0.01) and body protein retention increased (P < 0.05) with increasing fiber inclusion, while body fat retention increased numerically by 59%. The increase in BF was greatest for sows fed the high FS, intermediate when fed the medium strategy, and negligible for sows fed the lowest FS (P < 0.001). Nitrogen intake, N loss in feces, and N balance increased linearly, whereas N loss in urine tended to decrease with increasing inclusion of fibers in early gestation. Concomitantly, fecal energy output and energy lost as methane increased linearly (P < 0.001), while energy output in urine declined linearly. Total metabolizable energy (ME) intake therefore increased from 36.5 MJ ME/d in the low fiber group to 38.5 MJ ME/d in the high fiber group (P < 0.01). Changing the ME towards more ketogenic energy was expected to favor fat retention rather than protein retention. However, due to increased intake of ME and increased N efficiency with increasing fiber inclusion, the sows gained more weight and protein with increasing fiber inclusion. In conclusion, increased feed intake improved both fat and protein retention, whereas increased DF intake increased protein retention.

Oxidized myoglobin: Revealing new perspectives and insights on factors affecting the water retention of myofibrillar proteins

The impact of oxidized myoglobin (Mb) on myofibrillar protein (MP) oxidation and water retention was investigated. Results showed that the oxidation of Mb increased with increasing concentration of oxidized linoleic acid (OLA). In the presence of 100 mmol/L OLA, hemin iron decreased by 62.07 % compared to the control group. Further investigation showed that mild oxidation of Mb (≤10 mmol/L OLA) increased the water retention and the absolute value of the zeta potential of MP, whereas excessive oxidation (>10 mmol/L OLA) decreased these properties. With the increase of Mb oxidation, the carbonyl content in MP increased, and α-helices changed to random helix. And the tertiary structure changed. Pearson correlation analysis suggested that oxidized Mb affected the water retention of MP, which was closely related to hemin iron and non-hemin iron. In conclusion, OLA induced Mb oxidation, further promoted MP oxidation and affected its water retention.

Efficiency of protein combinations in diets for Rhamdia quelen: growth, digestive and metabolic biochemistry and nutrient deposition.

This study was conducted to determine the best combination of protein sources in diets for jundiá, based on growth, metabolism, and nutrient deposition. Five protein combinations were tested: casein + fish meal (control), casein + gelatin, casein + albumin, casein + albumin + fish meal, and albumin + fish meal, in diets containing 370 g Kg-1 of crude protein and 13.4 MJ Kg-1 of digestible energy. The fish (9.38 ± 0.12 g) were allocated in a water recirculation system at a density of 3.35 g L-1 per experimental unit and fed until apparent satiety for 40 days with the diets. The fish fed with the control diet had the highest final weight, specific growth rate, protein and feed efficiency ratio, protein retention, and best apparent feed conversion. On the other hand, fish that received casein + albumin and albumin + fish meal diets showed worse results in growth and body protein retention, low trypsin and chymotrypsin activity, and high intestinal amylase activity. Therefore, the combination referred to as control (casein + fish meal) conclusively provides the best rhythm for nutrient digestion and metabolism processes, enabling fish to reach greater growth and retention of body protein with low whole-fish fat content.

Valor nutritivo das folhas secas de Cratylia argentea na alimentação de coelhos em crescimento

Abstract Nutrition in rabbit farming accounts for about 70% of all costs; in this sense, C. argentea is an excellent alternative as a feed, due to its bromatological composition. Our goal is to evaluate the nutritional value of diets formulated with the dried leaves of C. argentea to feed growing rabbits. Twenty-eight white rabbits of the New Zealand breed, 55 days old, of both sexes, were used. Two treatments (n = 14, 7 males, and 7 females) were evaluated in a randomized block experimental design in a 2 x 2 factorial scheme. The control diet (CTRL) was formulated to meet the nutritional requirements of growing rabbits, and the test diet (DRY) was obtained by replacing 20% of the CTRL diet with dried leaves of C. argentea. The experimental period was comprised of 11 days, with seven days of adaptation and four days of total feces collection. The average daily gain, dry matter intake, feed conversion, and final body weight were not affected by replacing the CTRL diet with the dried leaves of C. argentea. The coefficient of dry matter digestibility showed better results in CTRL rabbits (female: 72.19%, male: 72.48%). Rabbits fed with the CTRL diet exhibited a higher consumption of crude protein compared to those fed with the DRY diet, however, these rabbits showed higher elimination and lower retention of crude protein. It is concluded that C. argentea presents values of 1,400.67 kcal per kg of dry matter, digestible energy, and 3.34% digestible protein per kg of dry matter.

Dietary supplementation with xylanase suppresses the antinutritional effect of nonstarch polysaccharides of flaxseed and increases bone strength in broiler chickens.

The aim of this study was to determine the effects of xylanase and flaxseed the performance of chickens, digesta viscosity, nutrient retention, fatty acid profile in muscle, tibia strength and interrelations of these factors in broiler chickens fed a wheat-based diet. Seven hundred and twenty one-day-old Ross 308 cockerels were assigned to four treatments according to the contents of flaxseed (0 and 80 g/kg) and xylanase (0 and 0.1 g/kg) in the diet. Xylanase significantly decreased the intake of feed (p < 0.001), decreased feed conversion (p < 0.001), and reduced mortality (p = 0.050). In addition, xylanase significantly increased the retention of all nutrients (p = 0.010 -<0.001) except crude fibre, the fat content in breast meat (p = 0.029) and liver (p = 0.019) and the concentration of polyunsaturated fatty acids (PUFAs) in meat (p = 0.002). Flaxseed supplementation did not influence performance but decreased the retention of dry matter (p = 0.016), crude protein (p = 0.012), organic matter (p = 0.016) and nitrogen-free extract (p = 0.008). Xylanase in combination with flaxseed increased the content of n-3 fatty acids in the breast meat (p = 0.006). The lowest n-6/n-3 ratio (p = 0.001) was detected in the flaxseed and flaxseed combined with xylanase groups. Significant interaction effects of flaxseed and xylanase on tibia strength (p = 0.030) and tibia ash content (p = 0.009) were detected. The administration of xylanase or flaxseed alone increased tibia strength. Compared with the control diet, the addition of flaxseed to the diet increased the digesta viscosity (p = 0.043) in the ileum, whereas the addition of xylanase decreased the level of this indicator. It can be concluded that xylanase is an enzyme suitable for increasing nutrient availability, and in the case of its addition to a flaxseed diet, it can reduce the antinutritional effect of flaxseed by reducing the viscosity of the digesta and increasing the content of health-promoting n-3 PUFAs.

Combined supplementation of hydroxyproline and vitamin C improved the growth and flesh quality of Pacific white shrimp (Litopenaeus vanname) cultured in low salinity water

This study investigated the effects of hydroxyproline (Hyp) and vitamin C (VC) addition to practical diet on growth and flesh quality of Pacific white shrimp (Litopenaeus vannamei) cultured in low salinity water. The control diet was formulated to contain 200 g/kg fish meal (FM20), and then 14 g/kg coated hydroxyproline (50%) (FM20H), 2 g/kg L-ascorbyl-2-phosphate (38%) (FM20C) and their combination (FM20HC) were added to the control diet to form four isoproteic (41.6%) and isolipidic (6.7%) diets. Pacific white shrimp with initial body weight of (1.50 ± 0.05) g were fed the above four diets in low salinity water (salinity 1‰) for 8 weeks. The results showed that the combined supplementation of Hyp and VC (FM20HC) increased the weight gain by 13.4%, and decreased the feed conversion ratio by 0.18 (P < 0.05), while the individual supplementation (FM20H and FM20C groups) just numerically promoted the growth (P > 0.05). In addition, the FM20HC group also showed higher protein and lipid retention than the FM20 group (P < 0.05). In flesh quality, the supplementation of Hyp (FM20H) increased total collagen and heat-soluble collagen contents, flesh chewiness and free flavor amino acids contents (P < 0.05). The supplementation of VC (FM20C) increased the contents of total collagen, heat-insoluble collagen and free flavor amino acids, and decreased flesh lactic acid, malondialdehyde contents and steaming loss (P < 0.05). The combined supplementation of Hyp and VC (FM20HC) showed higher total collagen and heat-insoluble collagen contents than the individual supplementation (FM20H and FM20C) (P < 0.05), and the myofiber density, flesh hardness and shear force in the FM20HC group were also significantly higher than those in the control (P < 0.05). In summary, the combined supplementation of hydroxyproline and VC in practical diet with fishmeal content of 200 g/kg significantly improved the growth performance and flesh quality of L. vannamei cultured in low salinity water.

Effects of Substituting Fish Meal by Chlorella Meal in Practical Diet on Growth, Feed Utilization, and Flesh Quality of Pacific White Shrimp (Penaeus vannamei)

This study investigated the effects of substituting fish meal (FM) in practical diet by chlorella meal on the growth, feed utilization, and flesh quality of Pacific white shrimp (Penaeus vannamei). First, a basal diet was prepared with 200 g/kg FM inclusion (FM‐20), and then chlorella meal was used to reduce FM inclusion to 150 g/kg (FM‐15), 100 g/kg (FM‐10), 50 g/kg (FM‐5), and 0 g/kg (FM‐0), corresponding to the replacement levels of 25%, 50%, 75%, and 100% of dietary FM, respectively. Shrimp (1.37 ± 0.10 g) were fed with the five isonitrogenous and isolipidic diets for 56 days. No significant difference was observed in feed conversion ratio (FCR) and weight gain (WG) between FM‐20 and FM‐15 group (P &gt; 0.05), but when chlorella meal substituted 50% of dietary FM, WG, protein and lipid retention, and n‐3/n‐6 PUFAs in flesh were significantly reduced with significant increase in FCR (P &lt; 0.05). Survival, feed intake, meat yield, apparent digestibility coefficient of crude protein, dry matter, and flesh shear force showed no significant difference between FM‐20 and substituted groups (P &gt; 0.05). When dietary FM was totally substituted by chlorella meal, the body yellowness and redness and essential amino acid content in flesh, including Lys and Met, were significantly reduced (P &lt; 0.05). No significant differences were found in flesh total collagen, crude lipid, crude protein, serum biochemical indexes, flesh texture profiles (hardness, springiness, etc.), water holding capacity, antioxidant capacity, fatty acid, free amino acid composition, and muscle fiber density among the five treatments (P &gt; 0.05). To sum up, in a practical diet with 200 g/kg FM inclusion, chlorella meal successfully replaced 25% of dietary FM without adverse impacts on the growth and feed utilization, and the substitution of 75% of dietary FM did not negatively affect the flesh quality of Pacific white shrimp.

Concentration and desalting of Tetraselmis suecica crude extract by ultrafiltration

Downstream processing, encompassing molecule extraction and extract purification, is a critical step in microalgal biorefinery. This study focuses on the concentration and desalting of Tetraselmis suecica crude extract, which contains proteins, neutral carbohydrates, uronic acids, and pigments. The biomass was initially disrupted with a high-pressure homogenizer operating in moderate conditions (P = 300 bars and 2 passes). The liquid extract obtained was then desalted and concentrated by stirred-cell ultrafiltration. Two membranes, both made of polyethersulfone (PES) but with different molecular weight cut-offs (10 kDa and 30 kDa), were tested for this purpose. The filtration process lasted 3.15 ± 0.34 h, with the temperature maintained at 28 ± 3ºC. There was, therefore, a compelling need to reduce the ash content to facilitate valorization of the extracted proteins. Both membranes displayed time-dependent decreases in permeate flux, membrane permeability, and shear rate. The protein concentration of the permeate increased steadily over time, whereas the concentrations of ash and uronic acids remained constant during ultrafiltration. These results demonstrated the efficacy of both membranes for desalting the extract. After disruption, the ash content in the extract was initially high, at 35.5 ± 0.8% dry weight (DW), decreasing protein purity to 26.1 ± 0.3% DW. The 10 kDa membrane displayed superior molecule retention, resulting in an increase of protein concentration to 50 g.L-1 in the final retentate. The 10 kDa membrane eliminated 79.5 ± 0.5% of salts from the extract, potentially achieving the complete retention of proteins, pigments, and uronic acids; approximately 21.4 ± 3.6% of total carbohydrates were removed by this membrane.

Retention of Histidine-Rich Protein II Rapid Diagnostic Tests in Pregnant Women after Antimalarial Treatment

Malaria is a preventable and curable disease and malaria diagnosis is critical to reduce malaria burden. This study is aimed at investigating the persistent positivity of Histidine-Rich Protein II (HRP2) rapid diagnostic test in pregnant women after antimalarial treatment. The study was carried out in Turai Yar’adua Maternity and Children’s Hospital Katsina and Federal Medical Centre Katsina. Blood samples were collected with the help of trained physicians. The result from this study shows that HRP2 persist more in the blood of pregnant women after treatment with fensider. It was observed that in all the age groups, day 3, day 7 and day 14 post drug treatment, were still positive for malaria using HRP2 malaria rapid diagnostic tool. The HRP2 was positive for all the different age group after 21 and 28 days post treatment except the age group 30-34 (97.5%). The percentage in the positivity result of pregnant women testing positive after treatment with malaria drug changed at 35 and 42 days post treatment in almost all the age groups. There was no statistical significant difference between the different age groups (p&lt;0.05) in HRP2. The persistent positivity decreases in all the trimester at 35 and 42 days post treatment in HRP2. There was no statistical significant difference between the age of pregnancy p&gt;0.05. The highest negative result was observed in the second trimester in 42 days post treatment. This therefore provides a useful data for improved malaria diagnosis, treatment and guide for pregnant women who are susceptible to malaria parasite.

A bioinspired approach for the modulation of electroosmotic flow and protein-surface interactions in capillary electrophoresis using silylated amino-amides blocks and covalent grafting.

We explore a bioinspired approach to design tailored functionalized capillary electrophoresis (CE) surfaces based on covalent grafting for biomolecules analysis. First, the approach aims to overcome well-known common obstacles in CE protein analysis affecting considerably the CE performance (asymmetry, resolution, and repeatability) such as the unspecific adsorption on fused silica surface and the lack of control of electroosmotic flow (EOF). Then, our approach, which relies on new amino-amide mimic hybrid precursors synthesized by silylation of amino-amides (Si-AA) derivatives with 3-isocyanatopropyltriethoxysilane, aims to recapitulate the diversity of protein-protein interactions (π-π stacking, ionic, Van der Waals…) found in physiological condition (bioinspired approach) to improve the performance of CE protein analysis (electrochromatography). As a proof of concept, these silylated Si-AA (tyrosinamide silylation, serinamide silylation, argininamide silylation, leucinamide silylation, and isoglutamine silylation acid) have been covalently grafted in physiological conditions in different amount on bare fused silica capillary giving rise to a biomimetic coating and allowing both the modulation of EOF and protein-surface interactions. The analytical performances of amino-amide functionalized capillaries were assessed using lysozyme, cytochrome C and ribonuclease A and compared to traditional capillary coatings poly(ethylene oxide), poly(diallyldimethylammonium chloride), and sodium poly(styrenesulfonate). EOF, protein adsorption rate, protein retention factor k, and selectivity were determined for each coating. All results obtained showed this approach allowed to modulate the EOF, reduce unspecific adsorption, and generate specific interactions with proteins by varying the nature and the amount of Si-AA in the functionalization mixture.

Exploring the impact of turkey age on gut morphological parameters and nutrient utilisation

This study was carried out to determine the influence of turkey age on gut morphology and nutrient utilisation. Data on the gut morphology and nutrient utilisation were collected at the different age intervals of turkeys' growth: the starter (day-old - 8 weeks), grower (9 - 16 weeks), and finisher (17 - 24 weeks) phases. Nicholas turkeys were fed commercial turkey diets ad libitum for the eight weeks of each phase: starter, grower, and finisher. The еnеrgy utilisation paramеtеrs such as the efficiency of metabolisable energy used for energy retention (KRE), the efficiency of metabolisable energy used for lipid retention (KREF), the efficiency of metabolisable energy used for protein retention (KREP) were determined. Rеsults indicatе a significant incrеasе in NEP and REP ovеr timе, undеrscoring thе increasing еnеrgy dеmands associatеd with growth, protein and fat deposition. Results of the gut morphological paramеtеrs, еncompassing villus and crypt dimеnsions, muscular wall thicknеss, еpithеlial charactеristics, goblеt cеll sizеs, and thе ratio of villus hеight to crypt dеpth. Thеre was significant incrеasе (p&lt;0.05) in villus hеight, crypt dеpth, and еpithеlial thicknеss ovеr thе agе intеrvals, suggеsting an increase in thе absorptivе surfacе arеa and barriеr function of thе gut. The ratio of villus height to crypt dеpth furthеr indicatеs a maturation of thе gastrointеstinal tract. This study highlights the pivotal role of gut dеvеlopmеnt in shaping thе nutriеnt utilisation of turkеys.

Moderate replacement of fish oil with palmitic acid-stimulated mitochondrial fusion promotes β-oxidation by Mfn2 interacting with Cpt1α via its GTPase-domain

The mitochondrial matrix serves as the principal locale for the process of fatty acids (FAs) β-oxidation. Preserving the integrity and homeostasis of mitochondria, which is accomplished through ongoing fusion and fission events, is of paramount importance for the effective execution of FAs β-oxidation. There has been no investigation to date into whether and how mitochondrial fusion directly enhances FAs β-oxidation. The underlying mechanism of a balanced FAs ratio favoring hepatic lipid homeostasis remains largely unclear. To address such gaps, the present study was conducted to investigate the mechanism through which a balanced dietary FAs ratio enhances hepatic FAs β-oxidation. The investigation specifically focused on the involvement of Mfn2-mediated mitochondrial fusion in the regulation of Cpt1α in this process. In the present study, the yellow catfish (Pelteobagrus fulvidraco), recognized as a model organism for lipid metabolism, were subjected to eight weeks of in vivo feeding with six distinct diets featuring varying FAs ratios. Additionally, in vitro experiments were conducted to inhibit Mfn2-mediated mitochondrial fusion in isolated hepatocytes, achieved through the transfection of hepatocytes with si-mfn2. Further, deletion mutants for both Mfn2 and Cpt1α were constructed to elucidate the critical regions responsible for the interactions between these two proteins within the system. The key findings were: (1) Substituting palmitic acid (PA) for fish oil (FO) proved to be enhanced in reducing hepatic lipid accumulation. This beneficial effect was primarily attributed to the activation of mitochondrial FAs β-oxidation; (2) The balanced replacement of PA stimulated Mfn2-mediated mitochondrial fusion by diminishing Mfn2 ubiquitination, thereby enhancing its protein retention within the mitochondria; (3) Mfn2-mediated mitochondrial fusion promoted FAs β-oxidation through direct interaction between Mfn2 and Cpt1α via its GTPase-domains, which is essential for the maintenance of Cpt1 activity. Notably, the present research results unveil a previously undisclosed mechanism wherein Mfn2-mediated mitochondrial fusion promotes FAs β-oxidation by directly augmenting the capacity for FA transport into mitochondria (MT), in addition to expanding the mitochondrial matrix. This underscores the pivotal role of mitochondrial fusion in preserving hepatic lipid homeostasis. The present results further confirm that these mechanisms are evolutionarily conserved, extending their relevance from fish to mammals.

Changes in the germination and content of protein and gluten in the caryopses of hard and soft wheat under pulse pressure and prolonged storage conditions

The study aims to identify the aging characteristics of hard and soft wheat caryopses without treatment and following treatment with pulse pressure created by a shock wave. The grain was treated with pulse pressures of 11 and 29 MPa. After one month and after three years, the caryopses were sorted into vitreous and floury types. The germination of control soft wheat caryopses decreased after storage, while after pulse pressure treatment and storage, it exceeded that of the control. The germination of control hard wheat caryopses with floury endosperm decreased after storage to a greater extent than that of caryopses with vitreous endosperm. Pulse pressure treatment reduced the germination of caryopses with vitreous endosperm. No change was observed in the protein content of control hard wheat caryopses following prolonged storage. In this case, pulse pressure contributed to the reduction in protein content during storage. The protein content of soft wheat caryopses with floury endosperm decreased after storage. In these caryopses, pulse pressure treatment contributed to better protein retention. Thus, the caryopses of hard wheat and soft wheat with vitreous endosperm were less susceptible to aging as compared to the caryopses of soft wheat and caryopses with floury endosperm, respectively. Pulse pressure damaged the vitreous endosperm, accelerating seed aging. It also contributed to the long-term storage of soft wheat caryopses without a significant loss of viability. Thus, it is possible to treat soft wheat grain to extend its shelf life.