Zeitschriftenartikel zum Thema „Casein/whey solutions“

The incorporation of caseins and whey proteins into acid gels produced from unheated and heat treated skimmed milk was studied by confocal scanning laser microscopy (CSLM) using fluorescent labelled proteins. Bovine casein micelles were labelled using Alexa Fluor 594, while whey proteins were labelled using Alexa Fluor 488. Samples of the labelled protein solutions were introduced into aliquots of pasteurised skim milk, and skim milk heated to 90 °C for 2 min and 95 °C for 8 min. The milk was acidified at 40 °C to a final pH of 4·4 using 20 g glucono-delta-lactone/l (GDL). The formation of gels was observed with CSLM at two wavelengths (488 nm and 594 nm), and also by visual and rheological methods. In the control milk, as pH decreased distinct casein aggregates appeared, and as further pH reduction occurred, the whey proteins could be seen to coat the casein aggregates. With the heated milks, the gel structure was formed of continuous strands consisting of both casein and whey protein. The formation of the gel network was correlated with an increase in the elastic modulus for all three treatments, in relation to the severity of heat treatment. This model system allows the separate observation of the caseins and whey proteins, and the study of the interactions between the two protein fractions during the formation of the acid gel structure, on a real-time basis. The system could therefore be a valuable tool in the study of structure formation in yoghurt and other dairy protein systems.

SUMMARYThe effect of heating on plasmin activity in various media, including phosphate buffer pH 7·0, skim milk, blood plasma, solutions of casein and solutions of whey proteins were investigated. Plots of log residual activity υ. heating time were linear at all temperatures from 63 to 143 °C. In buffer solutions the presence of casein led to substantial substrate protection, the Arrhenius plots being linear both in the presence and absence of casein. The activation energy, Ea, for the inactivation reaction, was 62·4 kJ/mol in buffer alone and 58·4 kJ/mol with casein present at 25 mg/ml. In skim milk, despite the presence of casein at a similar concentration, plasmin was no more stable to heat than in buffer alone, and a curved Arrhenius plot was obtained indicating a more complex inactivation mechanism. Heating in the presence of proteins having free -SH groups accelerated the inactivation of plasmin. The role of -SH groups was confirmed by experiments with added α-lactalbumin, in which no free -SH groups occur, and reduced carboxymethylated β-lactoglobulin, both of which were without effect. In blood plasma, plasmin was less stable to heat than in buffer (pH 7·0) or in skim milk. Plasminogen behaved very similarly to plasmin either when activated to plasmin with urokinase before heating or when activated afterwards. A hypothesis is presented to describe the heat inactivation and denaturation of plasmin. Technologically important findings are that in skim milk plasmin was largely unaffected by pasteurization conditions and 30–40% of its activity remained even after ultra high temperature processing conditions.

Gelation is a significant operation in dairy processing. Protein gelation can be affected by several factors such as temperature, pH, or enzyme addition. Recently, the use of ultrasonication has been shown to have a significant impact on the formation of whey protein gels. In this work, the effect of ultrasonication on the gelation of casein systems was investigated. Gels were formed by the addition of 7·6 mm Tetra Sodium Pyro Phosphate (TSPP) to 5 wt% micellar casein (MC) solutions. Sonication at 20 KHz and 31 W for up to 30 min changed the surface hydrophobicity of the proteins, whereas surface charge was unaltered. Sonication before the addition of TSPP formed a firm gel with a fine protein network and low syneresis. Conversely, sonication after TSPP addition led to an inconsistent weak-gel-like structure with high syneresis. Gel strength in both cases increased significantly after short sonication times, while the viscoelastic properties were less affected. Overall, the results showed that ultrasonication can have a significant effect on the final gel properties of casein systems.

In deep processing of milk, microfiltration is used to isolate native micellar casein. The need to reduce its cost by increasing the efficiency of this process determines the relevance of research work in this area. The purpose of our research is to analyze the a priori information. This will determine the practical value and prospects of subsequent experimental determination of optimal parameters of the skim milk microfiltration process. The main steps of information search by keywords: selection of databases (Scopus, WOS, ScienceDirect, Googlescolar, etc.) and the most authoritative editions (J. of Dairy Science, J. Membrane Science, J. Membranes), where appearance of publications with practical application in the research subject is noted since 2007, bibliography analysis of scientific articles. Non-academic materials are excluded from the search because they lack full descriptions of research methods, which complicates the reproducibility of the presented results. Analysis of publications devoted to methods of increasing the efficiency of membrane separation of dairy raw materials showed that most of them are partial solutions to this problem. With the limitations - the properties of separation objects, membrane materials, types of apparatuses, etc. cause difficulties in the practical use of the results under changing physical and chemical characteristics of natural milk. But always the main operating parameters of the skim milk microfiltration process are the transmembrane pressure, the circulation rate of the separated system in the apparatus and its temperature. Optimal conditions of milk microfiltration for separation of native micellar casein should be sought experimentally on the basis of creating mathematical models of the process followed by their analysis by numerical methods, as the data given by the authors should be considered as indicative, depending on raw materials, membranes and separation technology.

SummaryA recently described nuclear magnetic resonance (NMR) method was evaluated for its usefulness in studying thermal effects on milk proteins. The increase in water proton T2 relaxation rate observed during thermal treatment of aqueous whey protein solutions above the denaturing onset temperature paralleled results obtained with the standard Rowland (1938) method. The influence of milk constituants on NMR characteristics was analysed. The NMR response increased with the ionic strength and the addition of caseinate or casein micelles. The relevance of the T2 relaxation probe for studying thermal modifications of milk proteins is discussed. It is proposed to apply the NMR method for determining either reversible or irreversible thermal denaturation of whey proteins in model Systems.

The aggregation of proteins after heating of calcium-fortified milks has been an ongoing problem in the dairy industry. This undesirable effect restricts the manufacture of calcium rich dairy products. To overcome this problem, a completely new approach in controlling the heat stability of dairy protein solutions, developed in our lab, has been employed. In this approach, high intensity, low frequency ultrasound is applied for a very short duration after a pre-heating step at ⩾70 °C. The ultrasound breaks apart whey/whey and whey/casein aggregates through the process of acoustic cavitation. Protein aggregates do not reform on subsequent post-heating, thereby making the systems heat stable. In this paper, the acid gelation properties of ultrasonicated calcium-enriched skim milks have also been investigated. It is shown that ultrasonication alone does not change the gelation properties significantly whereas a sequence of preheating (72 °C/1 min) followed by ultrasonication leads to decreased gelation times, decreased gel syneresis and increased skim milk viscosity in comparison to heating alone. Overall, ultrasonication has the potential to provide calcium-fortified dairy products with increased heat stability. However, enhanced gelation properties can only be achieved when ultrasonication is completed in conjunction with heating.

Micellar casein concentrate (MCC) is a high protein ingredient (obtained by microfiltration of skim milk) with an elevated level of casein as a percentage of total protein (TP) compared to skim milk. It can be used as an ingredient in cheese making. Feta-type cheese is a brined soft cheese with a salty taste and acid flavor. We theorize that Feta-type cheese can be produced from MCC instead of milk, which can improve the efficiency of manufacture and allow for the removal of whey proteins before manufacturing Feta-type cheese. The objectives of this study were to develop a process of producing Feta-type cheese from MCC and to determine the optimum protein content in MCC to make Feta-type cheese. MCC solutions with 3% (MCC-3), 6% (MCC-6), and 9% (MCC-9) protein were prepared and standardized by mixing water, MCC powder, milk permeate, and cream to produce a solution with 14.7% total solids (TS) and 3.3% fat. Thermophilic cultures were added at a rate of 0.4% to MCC solutions and incubated at 35 °C for 3 h to get a pH of 6.1. Subsequently, calcium chloride and rennet were added to set the curd in 20 min at 35 °C. The curd was then cut into cubes, drained for 20 h followed by brining in 23% sodium chloride solutions for 24 h. Compositional analysis of MCC solutions and cheese was carried out. The yield, color, textural, and rheological measurements of Feta-type cheese were evaluated. Feta-type cheese was also made from whole milk as a control. This experiment was repeated three times. The yield and adjusted yield of Feta-type cheese increased from 19.0 to 54.8 and 21.4 to 56.5, respectively, with increasing the protein content in MCC from 3% to 9%. However, increasing the protein content in MCC did not show significant differences in the hardness (9.2–9.7 kg) of Feta-type cheese. The color of Feta-type cheese was less white with increasing the protein content in MCC. While the yellowish and greenish colors were high in Feta-type cheese made from MCC with 3% and 6% protein, no visible differences were found in the overall cheese color. The rheological characteristics were improved in Feta-type cheese made from MCC with 6% protein. We conclude that MCC with different levels of protein can be utilized in the manufacture of Feta-type cheese.

Impacts of medium pH, temperature and coexisted proteins on the degradation of two flavonoids fisetin and quercetin were assessed by spectroscopic method in the present study. Based on the measured degradation rate constants (k), fisetin was more stable than quercetin in all cases. Increasing medium pH from 6.0 to 7.5 at 37?C enhanced respective k values of fisetin and quercetin from 8.30x10?3 and 2.81x10?2 to 0.202 and 0.375 h-1 (P<0.05). In comparison with their degradation at 37?C, fisetin and quercetin showed larger k values at higher temperature (0.124 and 0.245 h?1 at 50?C, or 0.490 and 1.42 h?1 at 65?C). Four protein products in medium could stabilize the two flavonoids (P<0.05), as these proteins at 0.10 g L-1 decreased respective k values of fisetin and quercetin to 2.28x10?2-2.98x10?2 and 4.37?10?2-5.97x10?2 h?1. Hydrophobic interaction between the proteins and the two flavonoids was evidenced responsible for the stabilization, as sodium dodecyl sulfate could destroy the stabilization significantly (P<0.05). Casein and soybean protein provided greater stabilization than whey protein isolate. It is thus concluded that higher temperature and alkaline pH can enhance flavonoid loss, whereas coexisted proteins as flavonoid stabilizers can inhibit flavonoid degradation.

Исследование структурообразования продуктов с промежуточной влажностью является важным элементом повышения качественных характеристик молочных консервов в странах, где данная категория пищевых продуктов широко внедрена в рацион питания населения. Определенную структуру консервированных продуктов, в том числе сгущенного молока с сахаром, обуславливают состав и свойства исходного сырья, технологический процесс производства и условия хранения. На всех этапах производства могут возникать пороки различного происхождения, ухудшающие комплексно показатели готового продукта. Для сгущенных молочных консервов с сахаром нерешенной проблемой является кристаллизация лактозы с образованием кристаллов размером более 15 мкм, детектируемых сенсорно. С начала века технологии производства существенно не изменились, для получения однородной консистенции в сгущенное молоко с сахаром принято вносить «затравку» лактозы. При этом не до конца изучено, как на процесс производства сгущенных молочных консервов будет влиять преобразование соотношения белковых компонентов в биосистеме. Одним из перспективных решений формирования заданных органолептических и физико-химических показателей сгущенных молочных систем, включая этап длительного хранения, является замена части традиционного белкового профиля нативными сывороточными белками, их концентратами или гидролизатами. В статье представлены результаты эксперимента по установлению влияния различных соотношений казеина и сывороточных белков на реологические характеристики сгущенного молока с сахаром в широком интервале рН. Самые высокие показатели динамической вязкости зафиксированы для всех экспериментальных образцов при рН 5,6, что объясняется интенсивным взаимодействием белковых частиц в концентрированных системах и гелеобразованием вследствие физико-химического воздействия. В 1,5-2 раза динамическая вязкость повышалась в системах с традиционным соотношением белковых фракций, где источником сывороточных белков выступали концентрат сывороточного белка и сыворотка подсырная сухая, что объясняется более интенсивным комплексообразованием k-казеина и b-лактоглобулина. Однако при доминировании в образце сывороточных белков из этих же источников наблюдалось снижение динамической вязкости во всем диапазоне рН. При введении в систему трансформированных в денатурирующих условиях сывороточных белков было зафиксировано повышение динамической вязкости в 1,5 раза при рН 5,6 и в 10-12 раз при рН 5,9 и 6,2. Полученные данные подтвердили потенциал детального изучения взаимодействий компонентов системы с целью получения необходимых показателей продукта, стабильных в процессе его хранения. This study of structure formation of products with intermediate moisture is important to improve the quality characteristics of canned milk in countries where this category of food products is widely introduced in the diet of the population. A certain structure of canned products, including sweetened condensed milk, is determined by the composition and properties of raw materials, technological production process and storage conditions. The defects of various origin can arise at all stages of manufacture, worsening indicators of a finished product in complex. Crystallization of lactose with formation of crystals larger than 15 microns by a sensory method is an open issue for canned condensed milk with sugar. Since the beginning of the century, production technology has not changed significantly; lactose «seed» is commonly added to condensed milk with sugar to obtain a homogeneous consistency. Thus, the influence of transformation of ratio of protein components in a biosystem on process of production of canned milk products is not studied up to the end. Replacement of a part of traditional protein profile by native whey proteins, their concentrates or hydrolysates is one of the most perspective solutions for forming of required organoleptic and physico-chemical parameters of condensed milk systems including the stage of long-term storage. The paper presents the results of the investigation of the influence of different ratios of casein and whey proteins on the rheological characteristics of condensed milk with sugar in a wide pH range. The highest values of dynamic viscosity were fixed for all experimental samples at рН 5.6, which can be explained by intensive interaction of protein particles in concentrated systems and gelation due to physical and chemical influence. Dynamic viscosity was 1.5-2 times higher in the systems with the traditional ratio of protein fractions, where whey protein concentrate and whey powder were the source of whey proteins. It can be explained by more intensive complexation of c-casein and b-lactoglobulin. However, a decrease in dynamic viscosity over the entire pH range was observed when whey proteins from the same sources dominated the sample. The introduction of whey proteins transformed under denaturing conditions recorded a 1.5-fold increase in dynamic viscosity at pH 5.6 and a 10-12-fold increase at pH 5.9 and 6.2. The obtained data confirmed the potential of a detailed investigation of the interactions of the system components in order to obtain the necessary product parameters stable during storage.

Consumption of garlic leads to the persistence of “garlic breath” due to the presence of malodorous sulfur volatiles which may persist for as long as 24 h. Therefore, the purpose of this study is to investigate the effect of yogurt and its components on the deodorization of garlic sulfur volatiles in breath and study the roles of these components in deodorization. Raw garlic was consumed with different treatments and at different times for breath analysis. Different components were mixed with the garlic for headspace analysis. Volatiles were measured using selected-ion flow-tube mass spectrometry. Consuming yogurt at the same time as garlic was more effective than consuming it before or after. Yogurt was the most effective at deodorization, followed by the emulsion, then protein or fat alone. Decreasing the pH of protein solutions increased deodorization because changes to the structure of the proteins exposed more binding sites for the volatiles, while decreasing the pH of water or fat had no effect on deodorization. Whey protein deodorized better than casein due to the presence of more cysteine binding sites for volatiles. This study proposes that the fat, protein, microbial culture, and water in yogurt have synergistic effects on the deodorization of garlic volatiles. This study’s findings can help in the development of novel products targeting sulfur volatiles, with broad applications for mitigating malodors produced by garlic.

The aim of this study was to examine the enzymatic hydrolysis of κ-casein by isolating and identifying the released peptides. The enzymes employed in the study were chymosin, plasmin and trypsin, as well as a cell-free extract from three Lactobacillus helveticus and nine Lactobacillus casei strains. The findings showed that the bond most sensitive to the proteolytic activity of chymosin was the Phe 105-Met 106. After 24 hours of hydrolysis a few other bonds in the casein macropeptide were also cleaved. Plasmin was found to have weak proteolytic activity under the conditions of this study. When the enzyme-substrate ratio was raised from 1:200 to 1:50, a few peptides were released from the N-terminal region. Trypsin was found to hydrolyze several κ-casein bonds, and peptides were released from almost all regions of the protein. The proteases of Lactobacillus had less effect than chymosin, plasmin or trypsin. The strains could be divided into three categories. L. helveticus strains had activity on bonds in the mid-section and C-terminal region, L. casei strains EB, P3, P8 and A 1 had activity on bonds in the N- and C-terminal regions, while L. casei A5 and M9 had activity only on bonds in the C-terminal region.

SummaryRaw skim milk was diluted 1000-fold using distilled water or various salt solutions as specified. Smooth, hyperbolic profiles of coagulation rate v. pH for casein were calculated from recordings of turbidity (400 nm) with time. The effects of pH, cation type, anion type and cleavage of k−casein by chymosin (EC 3.4.23.4) were determined. The maximum of pH-coagulation rate profiles decreased by 63, 85 and 94% when the skim milk diluent was changed from water to salt solutions of NaCl (100 mM), CaCl2 (50 mM) or MgCl2 (50 mM). The maximum of the pH–coagulation rate profile was 15 times greater when the Ca salt was changed from CaCl2 to Ca(SCN)2 (50 mM). The highest pH at which casein coagulation occurred increased from 4·45 to > 6·0 when Cu2+ (1 mM) was included with casein micelles dispersed in CaCl2 solution (50 mM). The addition of chymosin to casein micelles suspended in CaCl2 solution (70 mM) eliminated the inhibition of casein coagulation by Ca2+ at pH 4·5. It is proposed that ions such as Mg2+, Ca2+, and Na+, which generally associate with casein phosphate and carboxylate groups, increased the H+ concentration required to initiate the coagulation of casein, because H+ must displace bound Ca2+, Mg2+ or Na+ to reduce repulsive hydration forces between casein micelles, allowing attractive hydration forces (e.g. hydrophobic phenomena) to cause casein coagulation. Furthermore, it is proposed that ions such as Cl−, Br−, and SCN− bind to lysine, arginine and histidine groups and thereby decrease repulsive hydration forces between cationic casein micelles.

Protein ingestion is known to enhance post-exercise hydration. Whether the type of protein (i.e., whey, casein) can alter this response is unknown. Accordingly, this study aimed to compare the effects of the addition of milk-derived whey isolate or casein protein to carbohydrate-electrolyte (CE) drinks on post-exercise rehydration and endurance capacity. Thirty male soldiers (age: 24 ± 2.1 y; VO2max: 49.3 ± 4.7 mL/kg/min) were recruited. Upon losing ~2.2% of body mass by running in warm and humid conditions (32.3 °C, 76% relative humidity [RH]), participants ingested either a CE solution (66 g/L carbohydrate [CHO]), or CE plus isolate whey protein (CEW, 44 g/L CHO, 22 g/L isolate whey), or CE plus isolate casein protein (CEC, 44 g/L CHO, 22 g/L isolate casein) beverage in a volume equal to 150% of body mass loss. At the end of the 3 h rehydration period, a positive fluid balance was higher with CEW (0.22 L) compared to CEC (0.19 L) and CE (0.12 L). Overall mean fluid retention was higher in CEW (80.35%) compared with the CE (76.67%) and CEC trials (78.65%). The time of the endurance capacity test [Cooper 2.4 km (1.5 miles) run test] was significantly higher in CEC (14.25 ± 1.58 min) and CE [(12.90 ± 1.01 min; (p = 0.035)] than in CEW [(11.40 ± 1.41 min); (p = 0.001)]. The findings of this study indicate that the inclusion of isolate whey protein in a CE solution yields superior outcomes in terms of rehydration and enhanced endurance capacity, as compared to consuming the CE solution alone or in conjunction with isolate casein protein.

The interaction of κ-casein and β-lactoglobulin is fundamental to all heat-induced modifications of milk product functionality, such as the heat stability of concentrated milks. Purified native κ-casein B and β-lg A solutions were heated at 80 °C at pH 6·7 separately and in a mixture. The circular dichroism spectra in the near UV indicated irreversible changes in the disulphide bonding patterns involving both proteins. Alkaline- and SDS-PAGE of heated samples showed that, in the presence of κ-casein, less β-lg was converted into β-lg polymers and the rate of loss of native β-lg was greater. When κ-casein was added to previously heated β-lg and the mixture was heated, the κ-casein reacted with the heat-induced β-lg polymers more readily than with the β-lg native monomers. The formation of β-lg dimers, trimers etc. was diminished. It was concluded that, when β-lg and κ-casein were heated together, β-lg formed thiol-exposed monomers, which reacted with each other or with the native κ-casein depending on the relative concentrations of β-lg and κ-casein. The products of these reactions included some disulphide-bonded 1[ratio ]1 β-lg[ratio ]κ-casein complexes, some monomer κ-casein and a range of large aggregates held together by either or both disulphide bonds and hydrophobic association.

The pH-dependent dissociation and re-association behavior of casein proteins in alkaline environment was studied. In dilute aqueous solutions of pH = 12 and 13, casein was found to exist in the form of submicelles which represent the fundamental structural unit of casein micelles. Morphology and dimension of the casein submicelles were examined by scanning electron microscopy (SEM), transmission electron microscopy (TEM) and atomic force microscopy (AFM), revealing spherical particles of ~ 10 - 30 nm in diameter. When the pH was increased to 14, unexpected re-association of casein submicelles occurred, resulting in the formation of large network aggregates at pH > 14. Furthermore, calcium phosphate was found to be the main driving force behind the re-association, as verified by comparative experiments employing Ca-depleted casein. Based on an analysis of potential interactions between the proteins, this re-association is ascribed to decreased electrostatic repulsion, strengthened hydrophobic attraction and formation of calcium phosphate linkages between casein proteins.

Abstract Small micellar casein particles were formed in aqueous solutions of native casein after addition of polyphosphate. These so-called submicelles aggregated and gelled with a rate that increased with increasing temperature. The evolution of the viscosity during this process was determined at constant shear rate or shear stress. When applying a small shear stress the viscosity increased strongly until the shear rate became immeasurably slow, but when the applied shear stress exceeded a critical value (σc) the aggregates broke up and the viscosity reached a maximum. At longer times the viscosity decreased rapidly at first, followed by a very slow decrease. σc was independent of the shear rate and heating temperature, but increased strongly with increasing casein concentration. At constant shear rate the stress remained close to σc, but fluctuated irregularly. After cessation of shear flow, gels were formed rapidly. Oscillation shear measurements for σ > σc showed a strongly non-linear response at the time of maximum viscosity.

This research reveals the underlying mechanisms that make high-intensity ultrasound an effective tool to reduce the viscosity of micellar casein concentrates and to enhance the solubility of the respective powders. Micellar casein concentrates (MCC) gained great importance in the production of valuable food products with high protein content, but the processing properties of the reconstituted solutions are deficient. Even though several presumptions were established, the reasons why ultrasound is able to reduce the product viscosity and what limitations occur when using sonication technology are still not clear yet. Our study aims to investigate those reasons by combining analyses of viscosity measurements, particle size distributions, solubility, and hydration. The data presented demonstrate that undissolved, highly hydrated particles play an important role in micellar casein concentrates showing a high viscosity. We conclude on the high voluminosity of those particles, since improved solubility and decreased viscosity are accompanying effects. The determined voluminosities of those particles are 35–40% higher than for colloidal dissolved micelles. Hence, the viscosity reduction of up to 50% can be only obtained by sonicating micellar casein concentrates derived from powder reconstitution, whereas ultrasonication of freshly prepared membrane-filtrated MCC does not reduce viscosity.

The peptide Pro130–Thr–Ser–Thr–Pro–Thr–Ile–Glu–Ala–Val–Glu140–Ser–Thr–Val–Ala–Thr–Leu–Glu–Ala–Ser–Pro150–Glu–Val–Ile, which corresponds to residues 130–153 of κ-casein B, was synthesized and the conformation of the peptide in solution investigated by circular dichroism (CD) spectroscopy, structure prediction algorithms and 1H-nuclear magnetic resonance spectroscopy. In a solution containing the structure-enhancing solvent trifluoroethanol the CD spectrum was typical of a peptide in the α-helical conformation and nuclear magnetic resonance showed that the amino acids between Ile136 and Ser149 (κ-casein numbering) were predominantly in the α-helical conformation, but that Pro130 to Thr135 and Pro150 to Ile153 were not. In addition, Thr133–Pro134 and Ser149–Pro150 were primarily in the trans conformation, the residues from Thr131 to Thr135 were in unordered structures and the residues from Glu151 to Ile153 were in an extended conformation. Residues Glu137 to Glu140 and Thr145 to Ala148 also displayed some 310-helix character. When the peptide was dissolved in 10 mM-cetyltrimethylammonium chloride solution at pH 6, the CD spectra indicated that the proportion of helical structure was comparable to that of the peptide in trifluoroethanol solution (400 ml/l), whereas when the peptide was dissolved in buffer alone or in 10 mM-SDS solution, the CD spectra were consistent with a low helical content. Acidification of these solutions to pH 2·85 resulted in a slight increase in the helical content of the peptide in buffer and more markedly in buffer containing SDS. When the peptide was in 5 mM-CaCl2 solution at neutral pH, the CD spectrum indicated that some ordered structure was present. Taken together these results indicate that the ionizable residues Glu137, Glu140, Glu147 and Glu151 could be important in determining the stability of the putative helix. The structure predictions found that the sequence from Glu137 to Pro150 would be more likely to be in a helical than any other conformation in the intact bovine protein, but that pig, sheep and goat κ-caseins did not give a prediction of a strongly helical region in this part of the molecule.

Casein is a micellar protein rich in glutamic and aspartic acids as well as in phosphoserine. Considering its native affinity for calcium and the connection of sub-micelles through calcium phosphate nanoclusters, this protein holds promise for stimulating biomimetic mineralisation phenomena and direct binding with the mineral phase of hard tissues. In this work we prepared new hybrids based on casein embedded in a poly(2-hydroxyethyl methacrylate)-polyethyleneglycol diacrylate (PHEMA-PEGDA) hydrogel. The resulting materials were investigated structurally by Fourier transform infrared (FT-IR). Casein modified the water affinity and the rheological properties of the hybrids. The microstructure was explored by scanning electron microscopy (SEM) and the distribution of the protein was established by combined SEM micrographs and elemental mapping considering the casein-specific elements (P, N and S) not contained by the synthetic hydrogel matrix. The effect of casein on the mineralisation potential and stability of the mineral phase was investigated by FT-IR and SEM when alternating incubation in Ca/P solutions is performed. Increasing casein content in the hybrids leads to improved mineralisation, with localised formation of nanoapatite phase on the protein areas in the richest sample in protein. This behaviour was proved microstructurally by SEM and through overlapping elemental distribution of Ca and P from the newly formed mineral and P, S and N from the protein. This study indicates that nanoapatite-casein-PHEMA-PEGDA nanocomposites may be developed for potential use in bone repair and regeneration.

The purpose of this study was to compare four activation solutions (AS)—Woynarovich, Lahnsteiner, Kucharczyk, and Perchec—with the addition of 0.5% bovine serum albumin (BSA) for ide (Leuciscus idus) sperm activation and analysis with a CASA system. It was found that ide sperm can be activated using each AS within a pH range of 7.4–9.0 and an osmolality range of 160–200 mOsm kg−1. The effect of Woynarovich and Perchec solutions supplemented with BSA and casein at concentrations of 0.25, 0.5, 1.0, and 2.0% were also analyzed during the experiment. These two AS without protein supplementation (pure solutions) were the controls. Woynarovich and Perchec solutions supplemented with the minimum BSA concentration (i.e., 0.25%) significantly improved sperm motility (89.05% and 86.63%, respectively) compared to the controls (20.39 and 28.48%, respectively). Similar increases were also noted in progressively motile sperm (PRG, %), the curvilinear velocity of sperm (VCL, µm s−1), and the amplitude of lateral head displacement (ALH, µm). A similar trend in CASA parameters was also noted when casein was added to Woynarovich and Perchec solutions at a concentration of 0.25%. We concluded that 0.25% doses of each of the proteins were sufficient to prevent sperm adhesion to glass slides, and they can be used in research on ide sperm motility measurements.

Cheese whey management and disposal is a major issue for dairy industries due to its high level of chemical and biochemical oxygen demand. However, it can still represent a source of nutrients (i.e., sugars, proteins and lipids) that can be applied, among other options, as substrate for microbial growth. Yarrowia lipolytica can grow in different environments, consuming both hydrophilic and hydrophobic substrates, and tolerates high salt concentrations. In this work, the lipolytic and proteolytic profile of 20 strains of Y. lipolytica were tested on caseins and butter. Then, their growth potential was evaluated in four types of whey (caciotta, ricotta, squacquerone and their mix). Y. lipolytica showed a very strain-dependent behavior for both hydrolytic profiles and growth capabilities on the different substrates. The best growers for all the types of whey tested were PO1, PO2, and RO2, with the first one reaching up to 8.77 log cfu/mL in caciotta whey after 72 h. The volatile molecule profile of the samples incubated with the best growers were characterized by higher amounts of esters, acids, ketones and alcohols. In this way, cheese whey can become a source of microbial cultures exploitable in the dairy sector.

SummaryThe ethanol (EtOH) stability of skim milk and the stability towards aggregation of casein micelles diluted into ethanolic buffer solutions were compared using data obtained from previously published experiments. Differences in absolute stability and in relative response were observed when Ca2+ level and pH were adjusted, the buffer system results lying below those from skim milk in both cases. Increasing the ionic strength of skim milk adjusted to pH 7·0 lowered its EtOH stability whereas increasing the ionic strength of the diluting buffer increased the stability of the casein micelles. The hypothesis is put forward that the differences are due to the simultaneous precipitation of Ca phosphate when EtOH is added to skim milk. This draws calcium from the caseinate sites of the micelle, counteracting the destabilizing effects of the EtOH towards the micelle. Such removal and the consequent restructuring are kinetically controlled and micellar precipitation in skim milk finally occurs when the micellar coagulation time falls within the time scale of the restructuring reactions.

Casein and mucin have been shown to improve the erosion-protective properties of the pellicle when applied in combination. The aim of this in vitro study was to optimize the concentrations of these 2 proteins to achieve a maximum protective effect. For the 2 parts of this study, we prepared a total of 195 human enamel specimens and randomly assigned them to 13 groups, corresponding to 11 different casein-mucin concentration-combinations tested and 2 negative control groups (humid chamber). They underwent 5 cycles, consisting of pellicle formation from human whole saliva (2 h, 30°C), modification of the pellicle with casein and mucin in different concentrations (immersion in protein solutions for 2 h, 30°C), and erosion for 1 min in citric acid (0.65%, pH 3.5, 30°C). Surface microhardness (SMH), surface reflection intensity (SRI), and in the first part also calcium release were monitored during the cycling process, and analyzed with Kruskal-Wallis and post hoc Dunn’s tests. The results suggest that the best concentrations to achieve the highest erosion-protective effect are 3.0% casein and 0.81% mucin, which lead to a significant protection as measured by SMH as well as SRI compared to the unmodified pellicle. For the calcium release, no significant differences were found. This concentration combination corresponds to a general raise of the protein concentrations and a change in the molar ratio of the proteins as compared to earlier studies. Casein and mucin could now be incorporated at the determined concentration as natural ingredients in oral care products designed to protect against erosion.

Background: Mung bean is well known as a starch source, but the physiological effects of mung bean protein have received little attention. In this study, we isolated mung bean protein from de-starched mung bean solutions, and investigated its influence on lipid metabolism. Objective: The aim of this study is to clarify the influence of the lipid metabolism by consumption of mung bean protein isolate (MPI)Methods: Diets containing either mung bean protein isolate (MPI) or casein were fed to normal rats for 28 days.Results: Both groups ate the same amount of food, but the plasma triglyceride level, relative liver weight and liver lipid contents (cholesterol and triglyceride pool) in the MPI group were significantly lower than in the casein group. In the MPI group, the expression of sterol regulatory-element binding factor 1 (SREBF1) mRNA in the liver was significantly different when compared with the casein group. The significantly lower levels of insulin and free fatty acids in the MPI-fed rats may be due to the regulation of genes related to lipid metabolism in the liver.Conclusions: These results suggest that MPI may improve the plasma lipid profile by normalizing insulin sensitivity.Keywords: mung bean, Vigna radiata L., 8S globulin, triglyceride, β-conglycinin, 7S globulin, insulin sensitivity, SREBF1

Wine fining comprises any operation resulting in the cleaning, stabilisation and improvement of the organoleptic characteristics of wine. Fining agents, such as certain proteins of varying electrical charges, interact physicochemically with wine polyphenols in ways not yet fully understood with diverse effects on wine attributes. In this study, four widely used protein fining agents (casein, ovalbumin and two gelatins) were compared for their ability to interact with three different types of phenolic-rich solutions: a tannic acid solution, a wine grape seed extract and seven different varietal wines. A phenolic characterisation of both the seed extract and all the wines was carried out. The interaction between the protein fining agents and the phenolic solutions was evaluated by diffusion and precipitation tests on cellulose membranes and protein staining. All four protein fining agents interacted to differing extents by producing soluble and insoluble complexes with the tannic acid, grape seed extract and wines. In the wines, the interactions were found to not only depend highly on the protein composition of the fining agent, but also on the chemical composition of the wine. Notwithstanding, the observed phenolic differences between the wines could not fully account for the differential interactions between wine and the fining agents, thus indicating that the wine matrix as a whole may play a role in those interactions.

Introduction: Casein is a milk protein that has the same number of positive and negative charges in a pH around 4.7. This characteristic is called the isoelectric point (pI). The pI varies from protein to protein and it depends on the charges of the lateral chain of the constituent aminoacids. At this pH value the protein has its minimum solubility since the net charge is zero and the repulsion between molecules is decreased. Furthermore, the electrostatic interaction occurs between the protein molecules. Thus, they form clumps which tend to precipitate. On the other hand, when they are placed in a solution whose pH is above or below of its pI, the protein molecules have respectively a negative or positive net charge, with strong repulsion between themselves and great interaction with the solvent (water). Objective: This learning object presents a simulation of a laboratory practice for the determination of casein isoelectric point. Materials and Methods: Cartoons were planned in order to show the methodology procedures and biochemical fundamentals. Animations were developed with the aid of the Adobe ® Flash 8 software associated with logic programming. Results: The simulation consists of six steps that reproduce the activities performed in the laboratory. Among them, the user can observe different degrees of turbidity and / or precipitation of casein in solutions with different pH values, he can measure these values with paper indicator strips and then determine the pI of this protein. Conclusions: This learning object was tested by students of Biochemistry I, Pharmacy course since 2012/2. The navigation features, design, interaction, interactivity were considered excellent by 80% of students indicating that this object can be used as an interesting tool to assist the teaching and learning of basic biochemistry. Available at:www.ufrgs.br/gcoeb/PontoIsoleletricoDaCaseina/PontoIsoleletricoDaCaseina.swf

This study investigates the potential of Fourier transform infrared spectroscopy (FTIR) to monitor glycation-induced changes in protein structure. Aqueous solutions of sodium caseinate and glucose (1:2 w/w, pH 6·7) were heated at 90°C for 0, 10, 20, 40 and 60 min. Evidence for caseinate glycation was obtained by mass spectrometry techniques (electrospray (ESI) and matrix-assisted laser desorption ionisation (MALDI)). FTIR was able to discriminate between glycated and non-glycated sodium caseinate, when the data were analysed by multivariate statistical methods; principal component analysis (PCA) and soft independent modelling of class analogy (SIMCA). The techniques used were complementary and provided different levels of information about the glycated samples.

Glycine catabolism was studied in isolated rat liver mitochondria by measuring the release of 14CO2 from [1-14C]glycine. Mitochondria isolated from rats fed on a high-protein (60% casein) diet for 5 days showed an enhanced ability to catabolize glycine compared with mitochondria from rats fed on a normal-protein (15% casein) diet. Glycine catabolism was also stimulated in normal protein-fed rats if they ingested a single high-protein meal for 2 h before being killed, thus illustrating the rapid response of the glycine-cleavage system to protein intake. The stimulation of glycine catabolism in rats given a high-protein diet or meal was not evident if the mitochondria were incubated in the absence of P(i) (omitting ADP had no effect on the rate). Mitochondria from high-protein- and normal-protein-fed rats did not differ in their ability to accumulate glycine, a process which occurred far too rapidly to impose a limit on the rate of flux through the glycine-cleavage system. The stimulation of glycine catabolism by high-protein feeding was not associated with a change in mitochondrial matrix volume. Furthermore, mitochondria from rats fed on a high-protein meal maintained an enhanced ability to catabolize glycine compared with those from rats fed on a normal-protein meal when incubated in hypo-osmotic solutions of very low osmolarity. When mitochondria from high-protein- or normal-protein-fed rats were maximally activated by incubation in the presence of 0.25 microM-Ca2+, the rates of glycine catabolism were high, but similar, showing that the stimulation of glycine catabolism by high-protein feeding does not involve an increase in the total capacity of the system. These findings show that hepatic glycine catabolism is stimulated rapidly by high-protein feeding, a response that we suggest is involved in the disposal of the excess glycine in the diet.

SummarySeveral mathematical models are presented in an attempt to describe the kinetics of the enzyme-induced coagulation of casein micelles. In each model the primary phase of the clotting reaction is assumed to follow first order kinetics. The only differences amongst the various models centre on the definition of the flocculation rate constant, which is defined in seven different ways. The rate constants are defined and discussed in terms of activation energy and functionality theory. The first model is such that the number of functional sites is two. The second is such that the number is much larger. The third and fourth are such that there is an exponential energy barrier, one which has a magnitude proportional to the extent of proteolysis caused by the clotting enzyme. These two definitions differ only in the pre-exponent. In one case the pre-exponent is a constant, whereas in the other it is dependent on the size of clotting particles. The fifth and sixth definitions are also energy barrier rate constants, but the energy barrier changes in an arbitrary fashion with respect to time during proteolysis. The seventh definition assumes a large number of functional sites, but such that the number increases with extent of proteolysis. In the Payens nomenclature (Payens, 1989), all models could be considered to be ‘source’ models, and all are derived using the Drake moment equation (Drake, 1972). Only the first model has a truly constant flocculation rate parameter, and only this model has a relatively simple analytical solution. All other models yield analytical solutions only by way of infinite series expansions. Thus, all models are presented in terms of power series expansions, and only through the first five time-dependent coefficients. This confines all models to the early stages of coagulation. In all cases the first three coefficients are virtually the same. The first two coefficients involve only proteolysis, and the third includes initial flocculation information. Time-dependent changes in the flocculation rate constant begin to take effect in the fourth coefficient. When the fourth coefficients of the third and seventh models are compared, a simple relationship is suggested between free energy barrier removal and functional site generation, but only assuming that the number of functionalities is large.

The present study evaluated the antimicrobial in vitro effects of the salivary proteins lactoferrin and lysozyme on microorganisms involved in the carious process, obtaining their minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). Streptococcus mutans (ATCC 25175) and Lactobacillus casei (ATCC 7469) were submitted to broth macrodilution of lysozyme at 80 mg/mL and lactoferrin at 200 mg/mL. The tubes were read in a spectrophotometer after they had been incubated at 37 °C for 18 h, in a carbon dioxide chamber, in order to read the MIC. A new subculture was carried on agar plates to obtain the MBC. The agar diffusion method was also tested, using BHI agar with 100 µL of the standardized microbial inocula. Filter-paper disks soaked in 10 µL of the solutions lactoferrin (200 µg/mL) and lysozyme (80 µg/mL) were placed on the agar surface. Inhibition halos were not observed on the plates, showing the absence of the antimicrobial effects of these proteins in this method. The bactericidal and bacteriostatic effects of lysozyme on L. casei were 50.3 mg/mL and 43.1 mg/mL respectively. The bactericidal and bacteriostatic effects on S. mutans were 68.5 mg/mL and 58.7 mg/mL. Lactoferrin did not induce any inhibitory effects on any microorganism, even in the concentration of 200 mg/mL. There was not a synergic antimicrobial effect of proteins, when they were tested together, even in the concentration of 42.8 mg/mL of lysozyme and 114 mg/mL of lactoferrin (the highest values evaluated). S. mutans and L. casei were only inhibited by lysozyme, not affected by lactoferrin and by the synergic use of both proteins.

ABSTRACT Cell adsorption and selective desorption for separation of microbial cells were conducted by using chitosan-immobilized silica (CIS). When chitosan was immobilized onto silica surfaces with glutaraldehyde, bacterial cells adsorbed well and retained viability. Testing of the adsorption and desorption ability of CIS using various microbes such as Escherichia coli, Aeromonas hydrophila, Pseudomonas aeruginosa, Bacillus subtilis, Micrococcus luteus, Staphylococcus aureus, Staphylococcus epidermidis, Lactobacillus casei, Streptococcus mutans, Streptococcus sobrinus, Streptococcus salivarius, Saccharomyces cerevisiae, Saccharomyces ludwigii, and Schizosaccharomyces pombe revealed that most microbes could be adsorbed and selectively desorbed under different conditions. In particular, recovery was improved when l-cysteine was added. A mixture of two bacterial strains adsorbed onto CIS could also be successfully separated by use of specific solutions for each strain. Most of the desorbed cells were alive. Thus, quantitative and selective fractionation of cells is readily achievable by employing chitosan, a known antibacterial material.

That’s a summary of our research in Greece and Germany as it concerns with their “labor market”. We examine the 3-polar system in the labor market, State-Company-Citizen. The aim of this paper is to show the bargaining possibilities when there are three involved parties on a labor market and two of them are active decision-makers. The third one is stakeholder who does not directly take part in the decision-making process. We will show possible solutions for increasing the benefit for all three parties. As an introduction, basic statistical data from Greece and Germany will be presented and structured. After this, the different behaviors of the parties in both countries will be regarded and their bargaining success will be illustrated.

SummaryThe effects of decreasing pH and micellar calcium concentrations of reconstituted skim milk and caseinate solution were studied by 1H and 17O NMR spectroscopy. The proton transverse relaxation rate 1/T2 of skim milk decreased as the pH decreased, reaching a minimum at pH 5·3. However, as the pH fell sodium caseinate solution showed a continuous increase in 1/T2, with no minimum. Analysis of proton relaxation as a function of the interpulse time in the CPMG (Carr-Purcell-Meiboom-Gill) sequence demonstrated that both the proton exchange mechanism and ‘bound’ water contributed to proton relaxation in skim milk. The study of 17O relaxation rate as a function of pH confirmed the change in protein hydration upon acidification. Increasing the amount of EDTA showed that the proton transverse relaxation rate of skim milk decreased until a plateau was reached when the micellar calcium was totally solubilized. With excess EDTA the relaxation rates of skim milk and caseinate solution were identical. A strong correlation was also found between the pH dependent relaxation rate and the solubilization of micellar phosphorus as detected by 31P NMR. Together, these results suggested that aggregation of caseins by calcium and colloidal calcium phosphate is mainly responsible for the excess hydration in skim milk micelles.

Using seleno-compounds and telluric compounds is a practical approach for developing solutions against drug-resistant bacterial infections and malignancies. It will accelerate the search for novel treatments or adjuvants for existing therapies. Selenium and tellurium nanospheres can be produced by lactic acid bacteria. The bacteria can differentiate the selenium and tellurium when the medium contains both selenite and tellurite. Therefore, our question in this study was the following: are they making alloys from the selenium and tellurium and what will be the composition, color, and shape of the nanoparticles? We used a simple microbial synthesis to produce nanoselenium, nanotellurium, and their alloys from sodium selenite and sodium tellurite using Lactobacillus casei. This bacterium produced red spherical amorphous elemental selenium nanospheres with a diameter of 206 ± 33 nm from selenite and amorphous black nanorods with a length of 176 ± 32 nm and a cross-section of 62 ± 13 nm from tellurite. If the initial medium contains a mixture of selenite and tellurite, the resulting nanoparticles will contain selenium and tellurium in the same ratios in the alloy as in the medium. This proves that Lactobacillus casei cannot distinguish between selenite and tellurite. The shape of the nanoparticles varies from spherical to rod-shaped, depending on the ratio of selenium and tellurium. The color of nanomaterials ranges from red to black, depending on the percentage of selenium and tellurium. These nanomaterials could be good candidates in the pharmaceutical industry due to their antipathogenic and anticarcinogenic properties.

Thermo diffusion and chemical effects on heat transfer in MHD mixed convection flow and masstransfer past an infinite vertical plate with Ohmic heating and viscous dissipation have beenstudied. Approximate solutions have been derived for velocity, temperature, concentration profiles,skin friction, rate of heat transfer and rate of mass transfer using perturbation technique. Theobtained results are discussed with the help of graphs to observe the effect of various parameterslike Schmidt number (Sc), Prandtl number (Pr), Magnetic parameter (M),Soret number (So) andchemical parameter (K), taking two cases viz. Case I: when Gr > 0 (flow on cooled plate) and CaseII: Gr < 0 (flow on heated plate). Thermal diffusion causes both the fluid velocity and temperature tofall due to the presence of the chemical effect. Velocity and temperature profiles are higher formercury than electrolytic solution. Soret effect increased the concentration of the fluid while chemicaleffect decreased.Keywords: Chemical effect; thermo diffusion; magnetic field; heat-mass transfer.DOI: 10.3329/jname.v6i2.3761

A numerical investigation has been implemented to elucidate the effect of vertical and horizontal vibration at normal gravity on natural convection in a square enclosure filled with air at Rayleigh number 7×107 and 4× 108. The enclosure was comprised of two vertical and opposed surfaces (the right hot and the left cold) while the two other surfaces are adiabatic. The two-dimensional, low-Reynolds number k ? ???? turbulence model is applied to enable it to cope with low Reynolds number flows. By transforming the equation of (continuity, Navier-Stokes and energy) using finite volume method from differential forms to algebraic forms using SIMPLE algorithm with hybrid scheme dealing with the time term are adopted to solve the governing equations. A computer program in Fortran 90 was built to carry on the numerical solution. Three cases were studied in this work, case I(reaches to steady state and then begins the effect of vibration at each frequency), caseII and caseIII(begin the effect of vibration from the transient at ascending and descending frequencies respectively).After the validity of the present code by comparing results with these of previous study for similar conditions, solutions have been obtained for Prandtle number of 0.7, aspect ratio (A=1). In the high Rayleih number case (Ra=4×108), the gravitional thermal convection dominates, and the vibration motion does not enhances the heat transfer remarkably. In contrast, in low Rayleigh (Ra=7×107), the vibration thermal convection is dominant, and the vibration enhaces the heat transfer rate significantly. The effect of vertical directional vibration is more powerful in caseII(ascending frequency), when the horizontal directional vibration more effective in case III(descending frequency).

The contemporary methodology of business process modeling is closely tied to process mining. The aim of the study is to develop a method of creating business process models through the restoration of links between events recorded in logs in the absence of CaseID data based on graph neural networks. The problem is solved by applying the graph convolutional networks architecture. The study employs a combination of a weighted adjacency matrix and an adjacency matric accounting for the graph data structure. Textual information about the tasks involved in the business process is considered when implementing the feature matrix using embeddings. The Navec embedding model is chosen to represent task titles as numerical vectors. The study was based on parsing the technological log of the 1C:Enterprise system. The obtained solutions make it possible to restore the required connection (Sequence flow) in the model of the "Approval of a commercial offer" business process in the absence of Case ID data in the event log as part of the "Reset request" task.

To assess- the release of calcium and phosphate ions from a fissure sealant containing amorphous calcium phosphate (ACP), and to determine the re-release capacity of these ions when charged with a solution containing casein phosphopeptide-amorphous calcium phosphate (CPP-ACP). Nine blocks of ACP resin-based sealant were prepared and immersed in three solutions at different pH (4.0, 5.5, 7.0), and calcium and phosphate ion release was measured with ion chromatography at 1, 3, 5, 7, 14, 21 and 28 days after immersion. Sixty days after immersion, each block was charged with CPP-ACP solution in three 7-day cycles to investigate the re-release of these ions, which was measured on days 1, 3, and 7. No difference was observed in initial calcium ion release at pH 4.0 and pH 5.5. At both values, ion release was significantly higher than at pH 7.0 (p<0.001). Initial phosphate release was significantly different among the three pH values (p<0.001). After re-charging the specimens, calcium ion re-release was greater than phosphate ion release. Initial ion release from ACP resin-based sealant was greatest at the lowest pH. Ion release decreased with time. As the number of recharge cycles increased, ion re-release also improved. Phosphate ion re-release required more recharge cycles than calcium ion re-release.

Abstract The application of near infrared spectroscopy (NIRS) to high-moisture samples (i.e., silages) has shown that accuracy does not match that of dried materials. Examination of spectra does not indicate absorbance levels to be the problem. The objective of this effort was to determine the effect of water on the spectra of model compounds. Near infrared spectra were taken using a Digi-Lab FTS-65 Fourier transform spectrometer. Liquids were examined by transmission and solids by reflectance. Examination of organic acids, alcohols, ketones, amines, and amides showed that the presence of water causes shifts in spectral wavelengths not related to OH or NH groups. The most significant shifts were for alcohols and ketones (up to 10+ nm at 90% H2O) and least for acids. These peak shifts increased with increasing amounts of water and varied within individual spectra and among the compounds tested. As solids, sugars and amino acids had many sharp peaks in their spectra. As solutions, however, the sharp spectral features disappeared, resulting in large broad peaks. The spectra of polymers such as starch, cellulose, and casein did not appear to be significantly altered by the presence of water (0-50%, w/w), although they often appear to alter the water spectra. Variations in water content, physical state, and concentrations of components, when combined with these results, may help

Abstract Objectives were to determine the effects of continuously infusing glucose (GLU) or casein (CAS) at the terminal ileum on circulating metabolism and intestinal architecture in pigs. Crossbred gilts (81 ± 1 kg BW) fitted with ileocecal cannulas and jugular catheters were enrolled in 2 experimental periods (P). Period 1 (4 d) served for the collection of baseline measurements. At the beginning of P2 (4 d), pigs were assigned to 1 of 3 infusion treatments: 1) control (CTL; water; n = 7), 2) GLU (500 g/d; n = 6), and 3) CAS (300 g/d; n = 6). Water, GLU, and CAS solutions were continuously infused (125 mL/h) through the ileocecal cannula for the entirety of P2. Pigs were sacrificed following P2 and segments of jejunum, ileum, and colon were collected. Data were analyzed using the MIXED procedure of SAS. During P2, circulating glucose and non-esterified fatty acids were similar among treatments (P > 0.10). A tendency for a treatment × time interaction was observed for insulin as it decreased over time in CTL pigs, while it remained almost unchanged in GLU and CAS treatments during P2 (P = 0.07). Blood urea nitrogen (BUN) decreased in GLU relative to CTL pigs during P2 (21%; P = 0.01), whereas it gradually increased (76%; P < 0.01) from 6 to 96 h post-infusion in CAS pigs when compared to their CTL counterparts. No differences on fecal pH were observed across treatments (P = 0.62). Ileum villus height tended to increase in CAS relative to CTL pigs (P = 0.08). Similarly, ileum villus height:crypt depth was increased in CAS compared to CTL pigs (P = 0.02). Goblet cell area was similar across treatments but tended to decrease in the jejunum in GLU when compared to their CTL counterparts (P = 0.09). In conclusion, continuously infusing GLU or CAS into the terminal ileum differently altered BUN patterns, but surprisingly had little effects on intestinal morphology.

Chromatographic stationary phases with specific capturing phosphoproteins is widely used in biological sample pretreatment. However, when captured protein is released, it is required to change the pH of the mobile phase or to use an eluent. Usually, the mobile phase or eluent are salt solutions with high concentration and extreme pH or toxic organic reagents. In this situation, these reagents will destroy the activity and structure of phosphorylated proteins. In addition, the mobile phase after switching the column takes longer time to restore the balance, reducing the experimental efficiency. In order to solve the these problems, we introduce temperature-reponsive materials into the chromatographic stationary phase to achieve the capture and release of phosphorylated proteins by changing the temperature only, in which we use water as the mobile phase. This approach overcomes the drawbacks of traditional methods, and makes the separation process safe and simple. Based on the surface initiated Reversible Addition Fragmentation Chain Transfer Polymerization (SI-RAFT) method, silica@pNIPAAm-nanoTiO2, a kind of Metal Oxide Affinity Chromatography, was synthesized by the rapid introduction of functional groups. The synthesis of silica@pNIPAAm-nanoTiO2was confirmed by infrared and X-ray photoelectron spectroscopy. The grafting rate and the lowest critical temperature were measured by TG and DSC. The results showed that the material had qualified temperature-sensitive properties. The grafting conformation and mobile phase pH of the material were optimized before testing the properties and found that when the material grafting ratio was 10% -15%, the graft density was 30%, and the mobile phase pH was 6, it had the best separate effect. Finally, the material successfully achieved the capture and release of adenosine triphosphate and casein phosphopeptides.

The improvement of the pharmacological strategy of non-alcoholic fatty liver disease is based on the study of the effect of pharmaceutical preparations on the structure and function of the liver. The pathogenesis of steatohepatitis is complex and multifactorial, mainly involving genetic, metabolic and environmental factors. The purpose of the study was to characterize the structural and functional parameters of the liver when using the biologically active compound Angiolin for the correction of experimental steatohepatitis. An experimental study was performed on 110 sexually mature white male rats weighing 180-220 grams, which were kept on a standard diet of vivarium. All animals were divided into two groups: control (30 intact animals) and experimental (80 animals). A model of non-alcoholic steatohepatitis was created for all animals of the experimental group. They were kept on a hypercaloric diet with a high fat and high cholesterol content for 8 weeks. After that, part of the animals (10 rats) was withdrawn from the experiment by intrapleural administration of sodium thiopental (50 mg/kg) and the necessary biochemical and morphological studies were performed. Part of the animals (30 rats) was continued to be kept on a high-fat diet for 4 weeks and the biologically active compound Angiolin was administered (20 rats), and Rings-Locke solutions were administered to 10 rats. After the creation of the model, the other animals of the experimental group (40 rats) were transferred to a full-fledged standard semi-synthetic starch-casein diet, and the biologically active compound Angiolin was administered for 20 rats and Ringer-Locke solution for another 20 rats for 4 weeks. Macroscopic evaluation and description of the liver of animals was carried out after withdrawal under thiopental anesthesia. Statistical analysis of the results was carried out using the program “STATISTICA 8” using parametric and non-parametric methods for assessing the results. It was found that the use of the biologically active compound Angiolin once a day for 30 days can reduce cytolysis syndrome (reduce biochemical parameters such as ALT, AST, gamma-glutamyl transpeptidase), reduce cholestasis syndrome (decrease in alkaline phosphatase level), and normalize liver function, improves the morphological state of hepatocytes, which indicates the normalization of the structural and functional state of the liver.

Cisplatin (CDDP) stands out as an effective chemotherapeutic agent; however, its application is linked to the development of significant adverse effects, notably nephro- and ototoxicity. The human organic cation transporter 2 (hOCT2), found in abundance in the basolateral membrane domain of renal proximal tubules and the Corti organ, plays a crucial role in the initiation of nephro- and ototoxicity associated with CDDP by facilitating its uptake in kidney and ear cells. Given its limited presence in cancer cells, hOCT2 emerges as a potential druggable target for mitigating unwanted toxicities associated with CDDP. Potential strategies for mitigating CDDP toxicities include competing with the uptake of CDDP by hOCT2 or inhibiting hOCT2 activity through rapid regulation mediated by specific signaling pathways. This study investigated the interaction between the already approved cationic drugs disopyramide, imipramine, and orphenadrine with hOCT2 that is stably expressed in human embryonic kidney cells. Regarding disopyramide, its influence on CDDP cellular transport by hOCT2 was further characterized through inductively coupled plasma isotope dilution mass spectrometry. Additionally, its potential protective effects against cellular toxicity induced by CDDP were assessed using a cytotoxicity test. Given that hOCT2 is typically expressed in the basolateral membrane of polarized cells, with specific regulatory mechanisms, this work studied the regulation of hOCT2 that is stably expressed in Madin–Darby Canine Kidney (MDCK) cells. These cells were cultured in a matrix to induce the formation of cysts, exposing hOCT2 in the basolateral plasma membrane domain, which was freely accessible to experimental solutions. The study specifically tested the regulation of ASP+ uptake by hOCT2 in MDCK cysts through the inhibition of casein kinase II (CKII), calmodulin, or p56lck tyrosine kinase. Furthermore, the impact of this manipulation on the cellular toxicity induced by CDDP was examined using a cytotoxicity test. All three drugs—disopyramide, imipramine, and orphenadrine—demonstrated inhibition of ASP+ uptake, with IC50 values in the micromolar (µM) range. Notably, disopyramide produced a significant reduction in the CDDP cellular toxicity and platinum cellular accumulation when co-incubated with CDDP. The activity of hOCT2 in MDCK cysts experienced a significant down-regulation under inhibition of CKII, calmodulin, or p56lck tyrosine kinase. Interestingly, only the inhibition of p56lck tyrosine kinase demonstrated the capability to protect the cells against CDDP toxicity. In conclusion, certain interventions targeting hOCT2 have demonstrated the ability to reduce CDDP cytotoxicity, at least in vitro. Further investigations in in vivo systems are warranted to ascertain their potential applicability as co-treatments for mitigating undesired toxicities associated with CDDP in patients.

BackgroundThe experience of art offers an emerging field in healthcare staff development, much of which is appropriate to the practice of palliative care. The workings of aesthetic learning interventions such as interactive theatre in relation to palliative and end of-life care staff development programmes are widely uncharted.AimTo investigate the use of aesthetic learning interventions used in palliative and end-of-life care staff development programmes.DesignScoping review.Data sourcesPublished literature from 1997 to 2015, MEDLINE, CINAHL and Applied Social Sciences Index and Abstracts, key journals and citation tracking.ResultsThe review included 138 studies containing 60 types of art. Studies explored palliative care scenarios from a safe distance. Learning from art as experience involved the amalgamation of action, emotion and meaning. Art forms were used to transport healthcare professionals into an aesthetic learning experience that could be reflected in the lived experience of healthcare practice. The proposed learning included the development of practical and technical skills; empathy and compassion; awareness of self; awareness of others and the wider narrative of illness; and personal development.ConclusionAesthetic learning interventions might be helpful in the delivery of palliative care staff development programmes by offering another dimension to the learning experience. As researchers continue to find solutions to understanding the efficacy of such interventions, we argue that evaluating the contextual factors, including the interplay between the experience of the programme and its impact on the healthcare professional, will help identify how the programmes work and thus how they can contribute to improvements in palliative care.References. Economist Intelligence Unit. 2015Quality of Death Index Ranking palliative care across the world. https://www.eiuperspectives.economist.com/healthcare/2015-quality-death-index, (2013 accessed 09/01/2017). World Health Organisation.WHO Definition of Palliative Care. Geneva: WHO. 2009.. Department of Health.Equity and excellence: Liberating the NHS. London: The Stationery Office Ltd. 2010.. Neuberger J.More care, less pathway: a review of the Liverpool care pathwayhttps://www.gov.uk/government/uploads/system/uploads/attachment_data/file/212450/Liverpool_Care_Pathway.pdf,(2013, accessed 09/12/2015). The National Council for Palliative Care. Commissioning Guidance for Specialist Palliative Care: Helping to deliver commissioning objectives.http://www.ncpc.org.uk/sites/default/files/CommissioningGuidanceforSpecialistPalliativeCare.pdf, (2012, accessed 15/12/2015). Leadership Alliance for the Care of Dying People.One Chance to get it Right.https://www.gov.uk/government/uploads/system/uploads/attachment_data/file/323188/One_chance_to_get_it_right.pdf, (2014accessed 15/12/2015). Cavaye J and Watts J. An Integrated Literature Review of Death Education in Pre-Registration Nursing Curricula: Key Themes, International Journal of Palliative Care, 2014, Article ID 564619, 19 pages. Gibbins J, McCoubrie R. Forbes K. Why are newly qualified doctors unprepared to care for patients at the end of life?Medical Education2011; 45(4): 389–399.. Gillan PC, van der Riet PJ and Jeong S. End of life care education, past and present: A review of the literature.Nurse Education Today2014; 34(3): 331–342.. Holms N, Milligan S and Kydd A. ‘A study of the lived experiences of registered nurses who have provided end-of-life care within an intensive care unit’,International Journal Of Palliative Nursing2014; 20(11): 549-556.. Levack P. Palliation and the caring hospital – filling the gap.Journal of the Royal College of Physicians Edinburgh2014; 44: 98–102.. Parliamentary and Health Service Ombudsman.Dying without dignity.http://www.ombudsman.org.uk/reports-and-consultations/reports/health/dying-without-dignity#_ftn1, (2015, accessed 15/12/2015).. NHS England.Actions for End of Life Care: 2014-16. https://www.england.nhs.uk/wp-content/uploads/2014/11/actions-eolc.pdf, (2014, accessed 15/12/2015).. Thun MJ, DeLancey JO, Centre MM, Jemal A, and Ward E M. The global burden of cancer: priorities for prevention.Carcinogenesis2010;31(1), 100–110.. Crawford P, Brown B, Baker, C, Tishler, V and Abrams B.Health Humanities. London: Palgrave Macmillan, 2015.. Tolstoy N. 1897.What is Art? [Qu est-ce que l' art?]. Paris: Gallimard, 1971.. Chinn PL, Maeve MK, and Bostick C. Aesthetic inquiry and the art of nursing.Scholarly Inquiry for Nursing Practice1997; 11: 83–96.. Goldenberg G. Sarah Sheets Cook: the invisible nurse.The Academic Nurse1999; 16(1): 26–28.. Buckley J. Massage and aromatherapy massage: nursing art and science.International Journal of Palliative Nursing2002; 8: 276–280.. Gramling KL. Ice chips and hope: the coach’s story of caring art.International Journal for Human Caring2004; 8(2): 62–64.. Gramling KL. Sarah’s story of nursing artistry: they do it with joy.Journal of Holistic Nursing2006; 24: 140–142.. Ryan J. Aesthetic physical caring: valuing the visible.Nursing in Critical Care2004; 9: 181–187.. Mendes IAC. Cultivating the art of service.Revista Latino Americana de Enfermagem2005; 13(2): 135.. Wyngaarden JB and Smith LH.Cecil textbook of medicine.Philadelphia: WB Saunders, 1985.. Saunders, J. The practice of clinical medicine as an art and as a science.Med Humanities2000; 26:18-22.. Egnew, T. The Art of Medicine: Seven Skills That Promote Mastery.FamilyPractice Management.2014; 21(4): 25-30.. Funch BS. The psychology of art appreciation. Copenhagen: Museum Tusculanum Press, 1997.Perry M, Maffulli N, Willson S and Morrissey D. The effectiveness of arts-based interventions in medical education: a literature review. Medical Education2011; 45(2): 141-148.. Wilson C, Bungay H, Munn-Giddings, C and Boyce M. Healthcare professionals’ perceptions of the value and impact of the arts in healthcare settings: A critical review of the literature.International Journal of Nursing Studies2016; 56: 90-101.. Ousager J and Johannessen H. Humanities in undergraduate Medical Education: A Literature Review. Academic Medicine2010; 85(6): 988-98.. Fairbrother G, Cashin A, Mekki TE, Graham I and McCormack B. Is it possible to bring the emancipatory practice development and evidence-based practice agendas together in nursing and midwifery?FoNS 2015 International Practice Development Journal2015; 5(1) [4].. Levac D, Colquhoun H and O’Brien KK. Scoping studies: advancing the methodology. Implementation Science2010; 5: 1–9.. Arksey H and O’Malley L. Scoping studies: towards a methodological framework.International Journal of Social Research Methodology: Theory & Practice2005; 8: 19-32.. Rumrill P, Fitzgerald S and Merchant W. Using scoping literature reviews as a means of understanding and interpreting existing literature.Work2010; 35: 399-404.. Grant M and Booth A: A typology of reviews: an analysis of 14 review types and associated methodologies.Health Info Libr J2009, 26: 91-108.. Brien S, Lorenzetti D, Lewis S, Kennedy J and Ghali W: Overview of a formal scoping review on health system report cards.Implement Sci2010, 5:2.. Armstrong R, Hall BJ, Doyle J and Waters E. Scoping the scope of a cochrane review.Journal of Public Health2011; 33: 147–150.. Daudt HM, Van Mossel C and Scott SJ. Enhancing the scoping study methodology: a large, inter-professional team’s experience with Arksey and O’Malley’s framework.BMC Medical Research Methodology2013; 13: 48.. Braun, V. and Clarke, V. Using thematic analysis in psychology. Qualitative Research in Psychology 2006; 3 (2): 77–101.. RefWorks.RefWorks your online research management, writing and collaboration tool,2009.. Bettany-Saltikov J.How to do a systematic literature review in nursing: a step-by-step guide. Maidenhead: McGraw-Hill/Open University Press, 2012.. Davis K. Drey N. and Gould D. What are scoping studies? A review of the nursing literature.Int J Nurs Stud2009; 46(10): 1386-400.. Pawson R. Evidence-based policy: in search of a method.Evaluation2002; 8(2): 157-181.. Duffin C. “Raising Awareness to Support People with Dementia in Hospital”,Nursing Older People2013; 25(5): 14–17.. Skye EP, Wagenschutz H, Steiger JA and Kumagai AK. Use of interactive theatre and role play to develop medical students’ skills in breaking bad news,Journal of Cancer Education2014; 29(4): 704–708.. Baer AN, Freer, JP, Milling DA, Potter, WR, Ruchlin H and Zinnerstrom KH Breaking bad news: use of cancer survivors in role-playing exercises,Journal of palliative medicine 200811(6): 885–892.. Tait GR and Hodges BD Residents learning from a narrative experience with dying patients: a qualitative study.Advances in Health Sciences Education2013; 18(4): 727–743.. Jones A. Death, poetry, psychotherapy and clinical supervision (the contribution of psychodynamic psychotherapy to palliative care nursing),Journal of advanced nursing1997; 25(2): 238–244.. Shapiro J, Hunt L. All the world’s a stage: the use of theatrical performance in medical education.Med Educ2003; 37(10): 922–7. Robinson S. Holistic health promotion: Putting the art into nurse education.Nurse Education in Practice2007; 7(3): 173--180.. Shapiro J, and Cho B. Medical Readers’ Theatre: Relevance to Geriatrics Medical Education,Gerontology & Geriatrics Education2011; 32(4): 350--366.. Durgahee T. Reflective practice: nursing ethics through story telling”,Nursing ethics1997; 4(2): 135–146.. Reilly J, Trial J, Piver D and Schaff P. Using Theatre to Increase Empathy Training in Medical Students,Journal for Learning through the Arts2012; 8(1).. Inske ep S and Lisco S. Alternative Clinical Nursing Experience in an Art Gallery.Nurse Educator2001; 26(3): 117--119.. Thompson T, van de Klee D, Lamont-Robinson, C and Duffin W. Out of Our Heads! Four perspectives on the curation of an on-line exhibition of medically themed artwork by UK medical undergraduates”,Medical Education Online 2010; 15.. Hickey D, Doyle C, Quinn S, O’Driscoll P, Patience D, Chittick K and Cliverd A. Catching’ the concept of spiritual care: implementation of an education programme”,International journal of palliative nursing2008; 14(8): 396–400.. Deloney LA and Graham CJ. Wit: using drama to teach first-year medical students about empathy and compassion,Teaching & Learning in MedicineCatching’ the concept of spiritual care: implementation of an education 15(4): 247–251.. Hodges HF, Keeley AC and Grier EC. Masterworks of art and chronic illness experiences in the elderly,Journal of advanced nursing2001; 36(3) 389–398.. Marchand L and Kushner K. Death pronouncements: using the teachable moment in end-of-life care residency training,Journal of palliative medicine2004; 7(1) 80–84.. Beach WA, Buller MK, Dozier DM, Bulle DB and Gutzmer K. The Conversations About Cancer (CAC) Project: Assessing Feasibility and Audience Impacts From Viewing The Cancer Play,Health communication2014; 29(5): 462–472.. Begley A, Glackin M and Henry R. Tolstoy, stories, and facilitating insight in end of life care: Exploring ethics through vicarious experience,Nurse Education today2011; 31(5): 516–520.. Kumagai AK. Perspective: Acts of Interpretation: A Philosophical Approach to Using Creative Arts in Medical Education,Academic Medicine2012; 87(8): 1138--1134.. Özcan NK, Bilgin H and Eracar N. The Use of Expressive Methods for Developing Empathic Skills,Issues in Mental Health Nursing2011; 32(2): 131–136.. Tuxbury J, McCauley P and Lement W. Nursing and Theatre Collaborate: An End-of-Life Simulation Using Forum Theatre,Journal of Nursing Education,2012; 51(8) 462–5.. Yalden J, McCormack B, O’Connor, M and Hardy S, Transforming end of life care using practice development: an arts-informed approach in residential aged care,International Practice Development Journal2013; 3(2).. Sklar DP, Doezema D, McLaughlin S and Helitzer D. Teaching communications and professionalism through writing and humanities: reflections of ten years of experience,Academic Emergency Medicine2002; 9(11): 1360–1364.. Sperlazza E and Cangelosi PR. The Power of Pretend: Using Simulation to Teach End-of-Life Care,Nurse Educator2009; 34(6): 276--280.. Gillis C. “Seeing the difference”: An interdisciplinary approach to death, dying, humanities, and medicine.Journal of Medical Humanities2006;27(2): 105–115.. Donovan T and Mercer D. Onward in my journey: preparing nurses for a new age of cancer care,Cancer nursing2003; 26(5) 400–404.. Fogarty CT. Fifty-five word stories: “small jewels” for personal reflection and teaching,Family medicine2010; 42(6): 400–402.. Foster W and Freeman E. Poetry in general practice education: perceptions of learners,Family Practice2008;25(4) 294–303.. Lillyman S, Gutteridge R and Berridge P. Using a storyboarding technique in the classroom to address end of life experiences in practice and engage student nurses in deeper reflection,Nurse Education in Practice2011; 11(3): 179–185.. Frei J, Alvarez S and Alexander M. Ways of Seeing: Using the Visual Arts in Nursing Education,Journal of Nursing Education2010; 49(12): 672--676.. Sherman DW, Matzo ML, Pitorak E, Ferrell BR and Malloy P. Preparation and care at the time of death: content of the ELNEC curriculum and teaching strategies,Journal for Nurses in Staff Development2005; 21(3): 93–102.. Franklin M. Acting on dilemmas in palliative care,Nursing times2001; 97(49): 37–38.. Epner DE and Baile WF. Difficult conversations: teaching medical oncology trainees communication skills one hour at a time,Academic Medicine2014; 89(4): 578–584.. Shannon SE, Long-Sutehall T and Coombs M. Conversations in end-of-life care: communication tools for critical care practitioners,Nursing in critical care.2011; 16(3): 124–130.. Deci EL and Ryan RM.Intrinsic motivation and self-determination in human behaviour. New York: Plenum Press, 1985.. Wee B, Hillier R, Coles C, Mountford B, Sheldon F and Turner P. Palliative care: a suitable setting for undergraduate interprofessional education,Palliative Medicine2001; 15: 187–492.. Meng AL and Sullivan J. Interactive theatre: an innovative conflict resolution teaching methodology,Journal for Nurses in Staff Development2011; 27(2): 65–68.. Salas R, Steele K, Lin A, Loe C, Gauna L and Jafar-Nejad P. Playback Theatre as a tool to enhance communication in medical education.Medical Education Online2013; 18(10).. Jonas-Simpson CF, Pilkington B, MacDonald C and McMahon E. Experiences of Grieving When There Is a Perinatal Death,Sage open2013.. Razavi D, Delvaux N, Marchal S, Durieux JF, Farvacques C, Dubus L and Hogenraad R. Does training increase the use of more emotionally laden words by nurses when talking with cancer patients? A randomised study,Br J Cancer2002; 87(1): 1–7.. Twigg R and Lynn M, Teaching End-of-Life Care Via a Hybrid Simulation Approach Simulation Approac,Journal of Hospice & Palliative Nursing2012; 14(5): 374–379.. Baile WF, Kudelka AP, Beale EA, Glober GA, Myers EG, Greisinger AJ, Bast RC, Goldstein MG, Novack D and Lenzi R. Communication skills training in oncology. Description and preliminary outcomes of workshops on breaking bad news and managing patient reactions to illness,Cancer1999; 86(5): 887–897.. Wilkinson S, Perry BK and Linsell L. Effectiveness of a three-day communication skills course in changing nurses’ communication skills with cancer/palliative care patients: randomised controlled trial,Palliative medicine2008; 22: 365–75.