Rapid Digestion Research Articles

The challenge to reduce crude protein contents of wheat-based broiler diets

The challenge to reduce crude protein (CP) contents of wheat-based broiler diets is both justified and formidable because the performance of broiler chickens offered reduced-CP, wheat-based diets is usually compromised. Moreover, broiler chickens offered wheat-based diets do not accommodate CP reductions as well as do those offered maize-based diets; this appears to stem from the higher protein concentrations and more rapid starch digestion rates of wheat. The higher protein concentrations of wheat than maize result in elevated inclusion levels of non-bound (synthetic, crystalline) amino acids (NBAA). This may be an impediment, because non-bound and protein-bound amino acids are not bioequivalent and intestinal uptakes of NBAA are more rapid than their protein-bound counterparts. This leads to post-enteral amino acid imbalances and the deamination of surplus amino acids, which generates ammonia (NH3). Because NH3 is inherently detrimental, it must be detoxified and eliminated as uric acid, which attracts metabolic costs. Moreover, inadequate NH3 detoxification may seriously compromise broiler growth performance. Also, consideration is given to some intrinsic wheat factors, including soluble non-starch polysaccharides, amylase–trypsin inhibitors and gluten, that may hold relevance. Several strategies are proposed that may enhance the performance of birds offered reduced-CP, wheat-based diets, including capping dietary starch:protein ratios, blending wheat with sorghum, whole-grain feeding in association with phytase, dietary inclusions of L-carnitine and the use of protected or slow-release amino acids. In future research, it should prove instructive to compare different wheats with a wide range of protein contents that, importantly, have been fully characterised for relevant parameters, to ascertain the most appropriate properties. The successful development and adoption of reduced-CP, wheat-based diets would be an enormous advantage for the Australian chicken-meat industry as it would diminish the huge dependence on imported, expensive soybean meal.

Real-time bottom-up characterization of protein mixtures enabled by online microdroplet-assisted enzymatic digestion (MAED).

Enzymatic digestion remains one of the "rate-determining steps" in the bottom-up analysis of proteins. However, by performing digestion in microdroplets generated from electrosonic spray, the reaction could be accelerated to a timescale lower than milliseconds. Here, we describe a simple and rapid online digestion platform named online microdroplet-assisted enzymatic digestion (MAED). It involves the integration of intact protein separation with enzymatic digestion in microdroplets. Via online MAED, various protein standards, including an antibody standard, were characterized in a bottom-up manner without prior digestion, and high sequence coverages were obtained. We further extended the application of online MAED to a more complex sample, mouse brain extract, where protein identifications were successfully yielded. Compared with the conventional bottom-up approach, a more comprehensive characterization could be obtained particularly for low molecular weight proteins. In short, we provide a rapid and alternative bottom-up analysis in a top-down fashion as well as a new possibility for microdroplet chemistry.

Sample preparation for structural mass spectrometry via polyacrylamide gel electrophoresis.

Mass spectrometry is an analytical technique that can detect protein molecules with high sensitivity. Its use is not limited to the mere identification of protein components in biological samples, but is recently being utilized for large-scale analysis of protein structures in vivo as well. Top-down mass spectrometry with an ultra-high resolution mass spectrometer, for example, ionizes proteins in their intact state and allows rapid analysis of their chemical structure, which is used to determine proteoform profiles. Furthermore, cross-linking mass spectrometry, which analyzes enzyme-digested fragments of chemically cross-linked protein complexes, allows acquisition of conformational information on protein complexes in multimolecular crowding environments. In the analysis workflow of structural mass spectrometry, prior fractionation of crude biological samples is an effective way to obtain more detailed structural information. Polyacrylamide gel electrophoresis (PAGE), known as a simple and reproducible means of protein separation in biochemistry, is one example of an excellent high-resolution sample prefractionation tool for structural mass spectrometry. This chapter describes elemental technologies for PAGE-based sample prefractionation including Passively Eluting Proteins from Polyacrylamide gels as Intact species for Mass Spectrometry (PEPPI-MS), a highly efficient method for intact in-gel protein recovery, and Anion-Exchange disk-assisted Sequential sample Preparation (AnExSP), a rapid enzymatic digestion method using a solid-phase extraction microspin column for gel-recovered proteins, in addition to presenting detailed experimental protocols and examples of their use for structural mass spectrometry.

A comparative investigation of anionic polysaccharides (sulfated fucan, ι-carrageenan, κ-carrageenan, and alginate) on the fabrication, stability, rheology, and digestion of multilayer emulsion

In this study, four important marine polysaccharides (sulfated fucan, ι-carrageenan, κ - carrageenan, and alginate) were selected and tailored to a similar molecular weight using heterologously overexpressed enzymes. These “standardized” polysaccharides were firstly used to elucidate the effect of anionic polysaccharides on the formation, stability, rheology, and digestion of the multilayer emulsions. Results showed that the four polysaccharides were all capable of adsorbing to the surfaces of lipid droplets at the pH values of 5, 4, and 3. ι-Carrageenan could adsorb to the surface of the droplet at a higher pH (pH 6) than the other three polysaccharides (pH 5). Stable multilayer emulsions were formed at a higher polysaccharide concentration (0.05 wt%) for κ-carrageenan compared with sulfated fucan, ι-carrageenan, and alginate (0.04 wt%). The emulsion containing alginate was susceptible to flocculation at low pH 3 and polysaccharide concentration >0.1 wt%. Multilayer emulsions fabricated by ι-carrageenan were stable under all the examined NaCl concentrations (0–400 mM). Multilayer emulsion fabricated by sulfated fucan showed less aggregation in the stomach stage, which is a good choice to encapsulate acid-intolerant substances. Multilayer emulsions fabricated by κ-carrageenan and alginate showed higher apparent viscosity, and the rheological curves of multilayer emulsions fabricated FUC or ICA were very similar. Moreover, multilayer emulsions fabricated by κ-carrageenan showed the fastest digestion rate and the highest digestion extent. These findings would facilitate the targeted design of multilayer emulsions by selecting suitable polysaccharides. • Four polysaccharides were tailed to similar molar mass by a repertoire of enzymes. • Multilayer emulsion with targeted functions was fabricated using polysaccharides. • ι-Carrageenan can fabricate multilayer emulsion with high salt stability demand. • κ-Carrageenan can form multilayer emulsion with the demand of rapid digestion. • Emulsion contained sulfated fucan had less aggregation in the stomach stage.

Physicochemical and Technofunctional Comparison of Starch from Varieties of Native Potato (Solanum Phureja) with Commercial Starches

Due to the high genetic variability of the potato, the starch can exhibit different properties of interest to the industry. With the present investigation, it is intended to know these differentiating characteristics in comparison with commercial sources that can be useful in different sectors. Therefore, the objective of this research was to establish a comparison between potato starches extracted from native varieties (Solanum phureja) with commercial potato and corn starches. A physicochemical analysis was carried out, for which characteristics such as amylose content, phosphorus, resistant starch (RS), slow digestibility starch (SDS), rapid digestibility starch (RDS), granule shape, granule size and crystallinity were evaluated. The technofunctional analysis included variables such as amylographic, thermogravimetric and differential scanning calorimetry analysis. The starches of the native varieties exhibited differential characteristics with respect to the commercial starches. Andina variety obtained the highest peak viscosity (8714 cP), followed by Ratona Blanca starch (6148 cP). In addition, these two varieties had the highest content of resistant starch (33.14±0.8) % and (28.9±0.7) % respectively), higher content of amylose (33.9 % and 35.5 % respectively), phosphorus (>0,08 % for both varieties), high crystallinity and higher enthalpy values (> 19 J/g for both varieties), which make them materials of potential use in different industries. The correlations presented between variables such as resistant starch (RS), enthalpy and crystallinity, showed possible differences at the structural level, which highlighted the differential characteristics of each starch.

Lutein-Fortified Plant-Based Egg Analogs Designed to Improve Eye Health: Formation, Characterization, In Vitro Digestion, and Bioaccessibility.

Lutein is a carotenoid found in real eggs that has been reported to have beneficial effects on eye health by reducing the risk of age-related macular degeneration. However, lutein is not often included in plant based (PB) egg analogs. It would, therefore, be advantageous to fortify PB eggs with this health-promoting carotenoid. Moreover, lutein is a natural pigment with a bright red to yellowish color depending on its concentration and environment. It can, therefore, also be used as a plant-based pigment to mimic the desirable appearance of egg yolk. Some of the main challenges to using lutein as a nutraceutical and pigment in PB foods are its poor water-solubility, chemical stability, and bioavailability. In this study, we encapsulated lutein in oil-in-water emulsions, which were then utilized to formulate whole egg analogs. Ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) protein isolated from a sustainable plant-based source (duckweed) was used to mimic the thermally irreversible heat-set gelling properties of globular egg proteins, with the aim of obtaining a similar cookability and texture as real eggs. The lutein content (80 mg/100 g) of the egg analogs was designed to be at a level where there should be health benefits. The protein (12.5 wt.%) and oil (10 wt.%) contents of the egg analogs were selected to match those of real egg. The effects of oil droplet size and oil type on the bioaccessibility of the encapsulated lutein were examined using the INFOGEST in vitro digestion model. For the emulsions formulated with long chain triglycerides (LCTs, corn oil), lutein bioaccessibility significantly increased when the initial droplet diameter decreased from around 10 to 0.3 μm, which was attributed to more rapid and complete digestion of the lipid phase for smaller droplets. For medium chain triglycerides (MCTs), however, no impact of droplet size on lutein bioaccessibility was observed. A high lutein bioaccessibility (around 80%) could be obtained for both LCTs and MCTs emulsions containing small oil droplets. Thus, both types of oil can be good carriers for lutein. In summary, we have shown that lutein-fortified PB eggs with good digestibility and bioaccessibility can be created, which may play an important role in ensuring the health of those adopting a more plant-based diet.

Quantification of Cl, Br and I in Geological Materials by NH4F Digestion: Comparison with Rapid Acid Digestion

The lack of analytical techniques for halogens in geological materials is mainly due to the loss of analytes during sample preparation. This study describes a rapid bulk rock digestion method (NH4F digestion) for determination of the abundances of Cl, Br and I in geological materials by SF‐ICP‐MS. During high temperature (200–240 °C) digestion, NH3 released from the decomposition of molten NH4F can effectively prevent the loss of halogens released from geological samples. Chlorine, Br and I were not lost during NH4F digestion at 220 °C for 0.25–6 h. The limits of quantitation for NH4F digestion were 2.8, 0.018 and 0.003 μg g‐1 Cl, Br and I, respectively. Most results for halogens in geological reference materials by NH4F digestion were in agreement with their certified values, confirming that the high‐performance rapid bulk rock NH4F digestion has sufficient digestion capability to extract Cl, Br and I from rocks, sediments and soils. In comparison, results obtained following acid digestion showed that HNO3 + HF digestion could effectively extract Br and I from soil and sediment samples, and that HNO3 acid digestion is only suitable to use for the determination of Br and I in soil samples.

Mechanistic Understanding of the Effects of Pectin on In Vivo Starch Digestion: A Review

Obesity and type II diabetes are closely related to the rapid digestion of starch. Starch is the major food-energy source for most humans, and thus knowledge about the regulation of starch digestion can contribute to prevention and improved treatment of carbohydrate metabolic disorders such as diabetes. Pectins are plant polysaccharides with complex molecular structures and ubiquitous presence in food, and have diverse effects on starch digestion. Pectins can favorably regulate in vivo starch digestion and blood glucose level responses, and these effects are attributed to several reasons: increasing the viscosity of digesta, inhibiting amylase activity, and regulating some in vivo physiological responses. Pectins can influence starch digestion via multiple mechanisms simultaneously, in ways that are highly structure-dependent. Utilizing the multi-functionalities of pectin could provide more ways to design low glycemic-response food and while avoiding the unpalatable high viscosity in food by which is commonly caused by many other dietary fibers.

Intake of slow-digesting carbohydrates is related to changes in the microbiome and its functional pathways in growing rats with obesity induced by diet.

The main cause of insulin resistance in childhood is obesity, which contributes to future comorbidities as in adults. Although high-calorie diets and lack of exercise contribute to metabolic disease development, food quality rather than the quantity of macronutrients is more important than food density. The purpose of the present study was to examine the effects of changing the quality of carbohydrates from rapidly to slowly digestible carbohydrates on the composition of the gut microbiota and the profiles of the functional pathways in growing rats with obesity due to a high-fat diet (HFD). During the course of 4 weeks, rats growing on an HFD-containing carbohydrates with different digestive rates were fed either HFD-containing carbohydrates with a rapid digestion rate (OBE group) or HFD-containing carbohydrates with a slow digestion rate (OBE-ISR group). A non-obese group (NOB) was included as a reference, and rats were fed on a rodent standard diet (AIN93G). An analysis of gut microbiota was conducted using 16S rRNA-based metagenomics; a linear mixed-effects model (LMM) was used to determine changes in abundance between baseline and 4 weeks of treatment, and functional pathways were identified. Gut microbiota composition at bacterial diversity and relative abundance, at phylum and genus levels, and functional profiles were analyzed by integrating the Integrated Microbial Genomes (IMG) database. The groups showed comparable gut microbiota at baseline. At the end of the treatment, animals from the ISR group exhibited differences at the phylum levels by decreasing the diversity of Fisher's index and Firmicutes (newly named as Bacillota), and increasing the Pielou's evenness and Bacteroidetes (newly named as Bacteroidota); at the genus level by increasing Alistipes, Bifidobacterium, Bacteroides, Butyricimonas, Lachnoclostridium, Flavonifractor, Ruminiclostridium 5, and Faecalibaculum and decreasing Muribaculum, Blautia, and Ruminiclostridium 9. Remarkably, relative abundances of genera Tyzzerella and Angelakisella were higher in the OBE group compared to NOB and OBE-ISR groups. In addition, some microbiota carbohydrate metabolism pathways such as glycolysis, glucuronic acid degradation, pentose phosphate pathway, methanogenesis, and fatty acid biosynthesis exhibited increased activity in the OBE-ISR group after the treatment. Higher levels of acetate and propionate were found in the feces of the ISR group compared with the NOB and OBE groups. The results of this study demonstrate that replacing rapidly digestible carbohydrates with slowly digestible carbohydrates within an HFD improve the composition of the gut microbiota. Consequently, metabolic disturbances associated with obesity may be prevented.

Development and validation of a LC–MS/MS method for quantitation of recombinant human growth hormone in rat plasma and application to a pharmacokinetic study

Recombinant human growth hormone (rhGH) is a peptide comprising 191 amino acids, that is mainly used to promote the growth of children and plays an important antiaging role. In the present study, a simple and sensitive quantitation method for rhGH in rat plasma was established by LCMS/MS. After simple and rapid enzymatic digestion of the plasma sample, two suitable surrogate peptides (LFDNAMLR and FPTIPLSR) were selected for quantitative analysis. The results showed good linearity over calibration range 10–2000 ng/mL. The quality control (QC) accuracy ranged from −13.8 to 14.3%, and the accuracy of the lower limit of quantification (LLOQ) ranged from −12.9 to 19.0%. The intra-day and inter-day precision ranges for all QCs were 1.7–13.6% and 4.0–7.0%, respectively. The method was successfully applied to intravenous and subcutaneous pharmacokinetic studies in rats. In comparison with previously published methods, our method features simple sample preparation combined with a short sample processing time (3.5 h), wide linear range (10–2000 ng/mL), small plasma volume (35 μL), and LLOQ (10 ng/mL).

A two-stage digestion of whole murine knee joints for single-cell RNA sequencing

ObjectiveSingle-cell RNA sequencing (scRNA-seq) is a powerful technology that can be applied to the cells populating the whole knee in the study of joint pathology. The knee contains cells embedded in hard structural tissues, cells in softer tissues and membranes, and immune cells. This creates a technical challenge in preparing a viable and representative cell suspension suitable for use in scRNA-seq in minimal time, where under-digestion may exclude cells in hard tissues, over-digestion may damage soft tissue cells, and prolonged digestion may induce phenotypic drift. We developed a rapid two-stage digestion protocol to overcome these difficulties. DesignA two-stage digest consisting of first collagenase IV, an intermediate cell recovery, then collagenase II on the remaining hard tissue. Cells were sequenced on the 10x Genomics platform. ResultsWe observed consistent cell numbers and viable single cell suspensions suitable for scRNA-seq analysis. Comparison of contralateral knees and separate mice showed reproducible cell yields and gene expression patterns by similar cell-types. A diverse collection of structural and immune cells were captured with a majority from immune origins. Two digestions were necessary to capture all cell-types. ConclusionsThe knee contains a diverse mixture of stromal and immune cells that may be crucial for the study of osteoarthritis. The two-stage digestion presented here reproducibly generated highly viable and representative single-cell suspension for sequencing from the whole knee. This protocol facilitates transcriptomic studies of the joint as a complete organ.

Research on chemical oxygen demand based on laser Fluorescence-Raman spectroscopy

In this invited paper, a novel water chemical oxygen demand (COD) detection method based on laser spectroscopy is proposed, and the COD value is obtained according to the intensity ratio of laser Fluorescence-Raman signal produced by laser incident into water. Based on the laser Fluorescence-Raman ratio (LFRR) method, we design a portable water quality measurement system. In which, a 405 nm semiconductor laser is used as the light source. The laser is coupled into the delivering port of Y-type fiber and then transmitted into water. The generated Mie scattered light, water Raman light and corresponding fluorescence are received and transmitted into spectrometer through the detecting port of Y-type fiber, and the COD value can be obtained by analyzing the LFRR of water spectra. The mixed solution of sodium humate and glucose is used as the COD standard solution to calibrate the designed system. The experiment results show that the optimal volume ratio of the two solutions is 1:29. When the COD of standard solutions is in the 1–12 mg/L concentration range, a good linear relationship can be found between the COD value and LFRR value with a correlation coefficient of 0.974. In addition, the COD of natural water samples are measured with LFRR method, the results of which are consistent with COD values obtained by rapid digestion spectrophotometry. Meanwhile, experimental results prove that the COD detection method proposed in this paper has the advantages of high sensitivity, high precision, high detection speed and simplicity, which can be widely used in various water areas for real-time COD monitoring.

CuNCs modified with dual-ligand to achieve fluorescence visualization detection of Tin (Ⅳ)

In this study, polyvinylpyrrolidone (PVP) supported and cysteine (Cys) modified copper nanoclusters (Cys-PVP-CuNCs) with high fluorescence intensity have been synthesized and used to establish a simple and sensitive assay for the determination of trace Sn4+. The presence of Sn4+ can lead to fluorescence quenching based on the aggregation of PVP-CuNCs caused by the coordination of Sn4+ and oxygen atoms. For Cys-PVP-CuNCs, the ligand Cys can transfer electrons to the metal center of the cluster, resulting in strong fluorescence enhancement of CuNCs and intensification of fluorescence quenching after interaction with Sn4+, which is conducive to more sensitive visual detection. The proposed assay allows for the detection of Sn4+ in a linear range from 5 μg L−1 to 100 μg L−1, with a detection limit as low as 4.5 μg L−1 at a signal-to-noise ratio of 3. Moreover, under UV light, the obvious fluorescent color change can be observed by naked eyes at Sn4+ concentration ranging from as low as 5 μg L−1 to 200 μg L−1. Benefiting from the high sensitivity of this assay, satisfactory test results have been obtained when using only 10 mg of canned food samples with rapid digestion. The proposed Cys-PVP-CuNCs fluorescent probe shows excellent stability, simple operation, good selectivity, and high sensitivity, and thus it is expected to be used in the real-time field detection of Sn4+.

Processing of semolina, a wonder resource for resistant starch production: In vitro digestibility and biochemical evaluation

With the advent of modern technology, the utilization of residues obtained after food processing are being largely explored for commercialization. Semolina, a starch rich food ingredient is one of such byproducts of food processing that has not been yet vividly studied, although it is profusely used as an important ingredient in Indian cuisines. Rapid digestibility of most starch rich foods boosts up the blood glucose level. Thus, the present study put forward an attempt to curtail the rapid digestibility of starch rich semolina flour by increasing its resistant starch content through enzymatic process. The enzymatically modified semolina flour (MS) was compared with its native counterpart (NS) on grounds of their digestibility pattern, biochemical and functional properties. A rise in resistant starch content by 9.3 ± 1.6 %, amylose content by 10.9 ± 1.2 %, crystallinity by 10.4 % and the drop in readily digestible starch by 11.9 ± 1.4 % and oil absorption by 2.1 ± 0.3 g/g were observed in MS. These initial findings of the present study are interesting as the results showed elevated potential of the modified semolina flour to be used as functional ingredient in cuisines worldwide.

Octenylsuccinic anhydride group distribution in esterified maize starches with different granular structure and its effect on starch digestibility

This study aimed to investigate the distribution of octenylsuccinic anhydrid (OSA) groups in different maize starch granules and its effect on starch digestibility. OSA starches with two substitution degrees (DS) about 0.02 and 0.05 were prepared. Confocal Laser Scanning Microscopy and Confocal Raman spectra showed that OSA groups distributed mainly on the granules surface at low DS of all the starches. The content of OSA groups in center area increased with the increasing DS for A-type starch, while the content of OSA groups decreased from the surface to inner at both low and high DS for B-type starch. This demonstrated that OSA reagent could travel through the granule pores and channels to arrive at the cavity in the A-type starch. However, due to a dense internal structure for B-type starch, OSA groups could graft into the granule only by penetration. Analysis of X-ray Diffraction showed that the degree of crystallinity slightly decreased as the increasing DS of OSA starch. The content of rapidly digestible (RDS) and slowly digestible starch (SDS) decreased, but that of resistant starch (RS) increased in the raw OSA starches with the increasing DS (RDS: 23.0%, RDS: 8.3%, RS: 72.3% for the raw waxy maize starch with 0.05 DS). It indicated that the hydrolysis of the OSA starches was influenced by the distribution of OSA groups.

Chick Provisioning in Grey-Faced Petrel (Pterodroma gouldi) under Environmental Stress

Grey-faced Petrels (Pterodroma gouldi) are colonial burrowing seabirds predominantly nesting on offshore islands of the upper North Island of New Zealand. We studied their chick provisioning on Te Hāwere-a-Maki during two years of unfavourable warmer La Niña conditions in 2011 and 2013. We intensively monitored chicks in each year, weighing chicks every 12 h for 10 consecutive days to estimate meal sizes following chick provisioning and to estimate 12-hourly body mass loss as a function of time since last feeding. We found a quadratic relationship of body mass loss with time since last feeding, with rapid digestion of meals following provisioning followed by a period of fasting from five days post feeding as chicks waited an unknown and variable amount of time until their next meal. The rate of body mass loss did not depend on chick age nor body mass, and did not differ between years, but heavier chicks included in our study were more likely to successfully fledge, suggesting a legacy of adult provisioning prior to our study commencing. Our regular handling of chicks for monitoring has no discernible impact on parent provisioning compared to a set of control chicks. The mean estimates of 100-gram meal sizes and 10-day foraging trip durations are likely to be below the break-even point for this species.

AuNPs/CdS QDs/CeO2 ternary nanocomposite coupled with scrollable three-dimensional DNA walker mediated cycling amplification for sensitive photoelectrochemical miRNA assay

Herein, a novel ternary nanocomposite (AuNPs/CdS QDs/CeO2) with excellent photoelectrochemical (PEC) performance was synthesized as signal probe to construct a near-zero background biosensor for sensitive miRNA-182-5p detection, by integrating with a scrollable three-dimensional (3D) DNA walker mediated cleavage cycling amplification. Impressively, the formation and rolling of scrollable 3D DNA walker triggered by target could realize dynamic, rapid and specific digestion of hairpin DNA on electrode with the aid of Exonuclease III (Exo III), which thus exposed abundant binding sites for assembling stable DNA labeled AuNPs/CdS QDs/CeO2 nanoprobes. Thanks to the formation of type-II heterojunction (between CeO2 and CdS QDs) and Schottky junction (generated by CeO2 and AuNPs), an ideal photoelectric conversion efficiency accompanied with stunningly improved photocurrent was thus acquired for significantly improving the detection sensitivity. It turned out that the detection limit (LOD) of biosensor was ultralow (31 aM). Significantly, the proposed PEC biosensor would exhibit great potential for the composite as a splendid indicator and provide an avenue for constructing the sensing platform with excellent sensitivity and ultralow background.

LC-MS/MS method for the quantitation of serum tocilizumab in rheumatoid arthritis patients using rapid tryptic digestion without IgG purification

The quantitation of serum tocilizumab using liquid chromatography tandem-mass spectrometry (LC-MS/MS) method has not been widely applied in clinical settings because of its time-consuming and costly sample pretreatments. The present study aimed to develop a validated LC-MS/MS method for detecting serum tocilizumab by utilizing immobilized trypsin without an immunoglobulin G purification step and evaluate its applicability in the treatment of rheumatoid arthritis (RA) patients administered intravenously or subcutaneously with tocilizumab. The tocilizumab-derived signature peptide was deciphered using a nano-LC system coupled to a hybrid quadrupole-orbitrap mass spectrometer. The serum tocilizumab was rapidly digested by immobilized trypsin for 30 min. The chromatographic peak of the signature peptide and that of the internal standard were separated from the serum digests for a total run time of 15 min. The calibration curve of serum tocilizumab concentration was linear with a range of 2–200 μg/mL. The intra- and inter-day accuracy and relative standard deviation (RSD) were 90.7%–109.4% and <10%, respectively. The serum tocilizumab concentrations in the RA patients receiving intravenous and subcutaneous injections were 5.8–28.9 and 2.4–63.5 μg/mL, respectively. The serum tocilizumab concentrations using the current method positively correlated with those using the enzyme-linked immunosorbent assay, although a systematic error was observed between these methods. In conclusion, a validated LC-MS/MS method with minimal sample pretreatments for monitoring serum tocilizumab concentrations in RA patients was developed.

Effects of Extrusion on Starch Molecular Degradation, Order-Disorder Structural Transition and Digestibility-A Review.

Extrusion is a thermomechanical technology that has been widely used in the production of various starch-based foods and can transform raw materials into edible products with unique nutritional characteristics. Starch digestibility is a crucial nutritional factor that can largely determine the human postprandial glycemic response, and frequent consumption of foods with rapid starch digestibility is related to the occurrence of type 2 diabetes. The extrusion process involves starch degradation and order–disorder structural transition, which could result in large variance in starch digestibility in these foods depending on the raw material properties and processing conditions. It provides opportunities to modify starch digestibility by selecting a desirable combination of raw food materials and extrusion settings. This review firstly introduces the application of extrusion techniques in starch-based food production, while, more importantly, it discusses the effects of extrusion on the alteration of starch structures and consequentially starch digestibility in various foods. This review contains important information to generate a new generation of foods with slow starch digestibility by the extrusion technique.

Azerbaijan effects on Agrochemical Indications and Effect of Erosion Process on Plant Productivity

Taking all these into account, it is important to increase the fertility of the soil from the erosion process in Ismayilli and to prevent the washing of fodder crops from perennial herbs. The cultivation of these plants in the mountainous regions protects the slopes from the terrible erosion process and provides the animals with a strong fodder. It is proved by the results of the research that restoration of fertility and ecological balance of erosion lands and the implementation of soil-agro-technical measures to increase productivity are of great importance. Due to the application of these measures, I, as a result of improving the water and physical properties of the affected land, prevent surface water flows. In addition, the results of the study have been proven by the fact that, for certain reason, erosion and erosion hazards are most likely to be taken over by the sowing of perennial herbs. Thus, perennial herbs, in particular, accumulate the nitrogen atmosphere of the legumes, enriches the soil with organic matter, accelerates the formation of water-resistant granular - topical structure and improves its water-physical properties, which in turn facilitates the rapid digestion of foodstuffs.