Concentrations Of Trypsin Research Articles

An eco-friendly near infrared fluorescence molecularly imprinted sensor based on zeolite imidazolate framework-8 for rapid determination of trace trypsin

This report depicts an environment-friendly molecularly imprinted fluorescence sensor for rapid determination of trypsin based on new near infrared (NIR) fluorescence carbon dots and metal–organic frameworks. The novel NIR fluorescence carbon dots were facilely synthesized from green plants by one-step solvothermal treatment, and the imprinted sensor was prepared by sol–gel under mild condition. The imprinted NIR fluorescence sensor was characterized by Fourier-transform infrared spectrometry, energy disperse spectroscopy, scanning electron microscopy, X-ray diffraction and transmission electron microscopy. The results showed the introduction of zeolite imidazolate framework-8 can improve the fluorescence response sensitivity and response speed. Under optimized conditions, the fluorescence quenching of the imprinted NIR fluorescence sensor was linear relationship with the increasing trypsin concentration in the range of 0.05–9.0 ng/L with a detection limit of 8.8 pg/L. The developed imprinted fluorescence sensor can selectively recognize trypsin in 1.5 min, which was applied to rapidly and sensitively detect trypsin in serum and urine samples successfully with the recoveries of 95.3–104.1% and 97.7–105.2%, respectively. The as-prepared imprinted fluorescence sensor provides a viable and eco-friendly method for rapidly sensitive detection of trace bio-macromolecule in complex samples.

Ultrasensitive off-on-off fluorescent nanosensor for protamine and trypsin detection based on inner-filter effect between N,S-CDs and gold nanoparticles

Herein we develop a novel “off–on-off” fluorescent nanosensor for the ultra-sensitive detection of protamine and trypsin based on the as-prepared nitrogen and sulphur doped carbon dots (N,S-CDs) and gold nanoparticles (AuNPs). N,S-CDs were treated as fluorophore, while AuNPs were treated as fluorescence absorber. By experimental studies, the fluorescence quenching mechanismof N,S-CDs by AuNPs was inner-filter effect (IFE). Upon addition of protamine to the mixed aqueous of N,S-CDs/AuNPs, the electrostatic interaction and Au-N bond between protamine and AuNPs resulted in AuNPs aggregation, and the quenched fluorescence of N,S-CDs could be recovered. Protamine is rich in arginine and can be specifically hydrolyzed by trypsin, resulting in the deaggregation of AuNPs and further quenching the fluorescence of N,S-CDs. Thus, we can determine the concentration of protamine and trypsin by measuring fluorescenceintensity of N,S-CDs.Under the optimized conditions, the fluorescence intensity showed a good linear relationship with protamine concentration of 20–260 ng/mL with the detection limit of 4.7 ng/mL. Meanwhile, quantitative determination of trypsin in a range from 10 to 140 ng/mL with the detection limit of 4.3 ng/mL can be achieved in this fluorescent nanosensor. This fluorescent nanosensor also showed excellent selectivity, and we successfully applied it for the detection of protamine and trypsin in human urine samples. Based on the above findings, we conclude that this novel “off–on-off” fluorescent nanosensor has great potential in clinical diagnosis.

Effect of Cereal α-Amylase/Trypsin Inhibitors on Developmental Characteristics and Abundance of Digestive Enzymes of Mealworm Larvae (Tenebrio molitor L.).

Simple SummaryThe main nutritionally relevant proteins of Tenebrio molitor L. larvae are cereal proteins. Cereals contain α-amylase/trypsin inhibitors (ATIs) that interact with digestive enzymes and which may impair the growth of the larvae. Despite the existing work on the subject, there is still a lack of information regarding the effects of ATIs on the relative abundance of various enzymes in larvae. Our aim was therefore to undertake an assessment of the potential effects of ATIs on the growth parameters and digestive enzyme contents of T. molitor. The larvae were fed with cereal meals containing different levels of ATIs. The developmental characteristics were evaluated and finally an analytical method based on liquid chromatography with tandem mass spectrometry (LC-MS/MS) was developed to quantify the relative abundance of enzymes in the larvae. The results indicated an increase in pupation and significantly higher protein concentrations in larvae fed with wheat meals compared to those fed with sorghum meals. Patterns of specific α-amylase activity (in mM maltose/min/mg protein) were similar to those of amylase activity (in mM maltose/min) and the larvae fed on high-ATI-content meals exhibited an increased death rate, although the results were not always significantly consistent. The results of the LC-MS/MS analysis showed a decrease of about half of the relative content of α-amylase among the three proteases monitored, as well as an increase in dipeptidylpeptidase I and chymotrypsin, whereas trypsin remained constant. Therefore, these results indicate that meal composition has an effect on the expression of T. molitor digestive enzymes.The objective of this work was to investigate the potential effect of cereal α-amylase/trypsin inhibitors (ATIs) on growth parameters and selective digestive enzymes of Tenebrio molitor L. larvae. The approach consisted of feeding the larvae with wheat, sorghum and rice meals containing different levels and composition of α-amylase/trypsin inhibitors. The developmental and biochemical characteristics of the larvae were assessed over feeding periods of 5 h, 5 days and 10 days, and the relative abundance of α-amylase and selected proteases in larvae were determined using liquid chromatography tandem mass spectrometry. Overall, weight gains ranged from 21% to 42% after five days of feeding. The larval death rate significantly increased in all groups after 10 days of feeding (p < 0.05), whereas the pupation rate was about 25% among larvae fed with rice (Oryza sativa L.) and Siyazan/Esperya wheat meals, and only 8% and 14% among those fed with Damougari and S35 sorghum meals. As determined using the Lowry method, the protein contents of the sodium phosphate extracts ranged from 7.80 ± 0.09 to 9.42 ± 0.19 mg/mL and those of the ammonium bicarbonate/urea reached 19.78 ± 0.16 to 37.47 ± 1.38 mg/mL. The total protein contents of the larvae according to the Kjeldahl method ranged from 44.0 and 49.9 g/100 g. The relative abundance of α-amylase, CLIP domain-containing serine protease, modular serine protease zymogen and C1 family cathepsin significantly decreased in the larvae, whereas dipeptidylpeptidase I and chymotrypsin increased within the first hours after feeding (p < 0.05). Trypsin content was found to be constant independently of time or feed material. Finally, based on the results we obtained, it was difficult to substantively draw conclusions on the likely effects of meal ATI composition on larval developmental characteristics, but their effects on the digestive enzyme expression remain relevant.

Detection of Sub-Nanomolar Concentration of Trypsin by Thickness-Shear Mode Acoustic Biosensor and Spectrophotometry.

The determination of protease activity is very important for disease diagnosis, drug development, and quality and safety assurance for dairy products. Therefore, the development of low-cost and sensitive methods for assessing protease activity is crucial. We report two approaches for monitoring protease activity: in a volume and at surface, via colorimetric and acoustic wave-based biosensors operated in the thickness-shear mode (TSM), respectively. The TSM sensor was based on a β-casein substrate immobilized on a piezoelectric quartz crystal transducer. After an enzymatic reaction with trypsin, it cleaved the surface-bound β-casein, which increased the resonant frequency of the crystal. The limit of detection (LOD) was 0.48 ± 0.08 nM. A label-free colorimetric assay for trypsin detection has also been performed using β-casein and 6-mercaptohexanol (MCH) functionalized gold nanoparticles (AuNPs/MCH-β-casein). Due to the trypsin cleavage of β-casein, the gold nanoparticles lost shelter, and MCH increased the attractive force between the modified AuNPs. Consequently, AuNPs aggregated, and the red shift of the absorption spectra was observed. Spectrophotometric assay enabled an LOD of 0.42 ± 0.03 nM. The Michaelis–Menten constant, KM, for reverse enzyme reaction has also been estimated by both methods. This value for the colorimetric assay (0.56 ± 0.10 nM) is lower in comparison with those for the TSM sensor (0.92 ± 0.44 nM). This is likely due to the better access of the trypsin to the β-casein substrate at the surface of AuNPs in comparison with those at the TSM transducer.

Trypsin Inhibitor Assay: Expressing, Calculating, and Standardizing Inhibitor Activity in Absolute Amounts of Trypsin Inhibited or Trypsin Inhibitors

AbstractFor expressing trypsin inhibitor activity (TIA), trypsin units inhibited (TUI), trypsin inhibited, and trypsin inhibitors have been used. Although the last two units are preferred, their calculations in current practices require refinement. With the proposed AOCS method Ba 12a‐2020, four experiments were conducted, using four trypsin preparations having specific activity of 11,625, 12,602, 13,728, and 14,926 Nα‐benzoyl‐L‐arginine ethyl ester (BAEE) units/mg protein, respectively. Experiment 1 determined the relationship between absorbance at 410 nm (A410) and trypsin concentration. Experiment 2 involved assaying raw and heated soybeans, expressing TIA as TUI/mg sample and μg trypsin inhibited/mg sample, and determining conversion factors between the two units. Experiment 3 resembled Experiment 2 except for using purified soybean Kunitz inhibitor (KTI) and Bowman‐Birk inhibitor (BBI). Conversion factors determined correlated highly with trypsin‐specific activity (R2 = 0.9789). After standardizing against a reference trypsin having 15,000 BAEE units/mg protein, a standardized conversion factor of 0.03 A410 (1.5 TUI) = 1 μg trypsin inhibited was determined. It remained consistent regardless of trypsin specific activity, with or without inhibitors, and type of inhibitor samples. By using purified inhibitors (Experiment 3), conversion values between TUI and μg trypsin inhibitor and between μg trypsin inhibited and μg trypsin inhibitor could also be calculated, enabling expression of TIA in amounts of pure KTI, BBI or their equivalents. Furthermore, when the AOCS method was modified with half substrate concentration, half trypsin concentration or half both (Experiment 4), TIA values in TUI could change with modifications but values in mg trypsin inhibited (standardized) or trypsin inhibitor remained consistent.

An anionic tetraphenyl ethylene based simple and rapid fluorescent probe for detection of trypsin and paraoxon methyl

In this study, we have developed an aggregation induced emission (AIE) based simple, sensitive, rapid and convenient biosensor for detection of trypsin and organophosphorous pesticides. The biosensor is composed of tetra-anionic sulphonyl derivative of tetraphenylethylene (Su-TPE) and cationic polyelectrolyte protamine (PrS), assembled by electrostatic interactions without any complex process of integration. The AIE property of Su-TPE/PrS system has been studied, in detail, using different spectroscopic techniques, such as, ground-state absorption spectroscopy, steady-state and time-resolved emission spectroscopy. The AIE phenomenon of Su-TPE molecules induced due to Su-TPE/PrS supramolecular complex formation has been supported by studying Su-TPE/PrS system under various environmental factors. After establishing the AIE phenomenon of Su-TPE/PrS complex, the potential of the sensing platform, for trypsin detection, has been explored based on the principle of trypsin dependent PrS hydrolysis. The detection system is highly sensitive in the linear trypsin concentration range of 0–16 nM with detection limit of 0.22 nM. Furthermore, the sensing platform also shows excellent selectivity towards trypsin. More importantly, based on trypsin inhibition, Su-TPE/PrS platform is also responsive towards organophosphorous pesticide detection. The commendable performance of the Su-TPE/PrS biosensor will enhance future development of rapid detection of trypsin along with fast high throughput screening of organophosphrous pesticides.

Fast in-line bottom-up analysis of monoclonal antibodies: Toward an electrophoretic fingerprinting approach.

For their characterization and quality control, monoclonal antibodies are frequently analyzed at the bottom-up level to generate specific fingerprints that can be used to tackle post-translational modifications or ensure production consistency between lots. To circumvent time-consuming and labor-intensive off-line sample preparation steps, the implementation of integrated methodologies from sample preparation to separation and detection is highly valuable. In this perspective, capillary zone electrophoresis appears as a choice technique since the capillary can subsequently be used as a vessel for sample preparation and electrophoretic discrimination/detection of the reaction products. Here, a fast in-line methodology for the routine quality control of mAbs at the bottom-up level is reported. Simultaneous denaturation and reduction (pretreatment step) were conducted with RapiGest® surfactant and dithiothreitol before in-line tryptic digestion. Reactant mixing was realized by transverse diffusion of laminar flow profile under controlled temperature. In-line digestion was carried out with a resistant trypsin to autolysis. The main parameters affecting the digestion efficiency (trypsin concentration and incubation conditions) were optimized to generate mAb electrophoretic profiles free from trypsin interferences. An acidic MS-compatible BGE was used to obtain high resolution separation of released peptides and in-line surfactant cleavage. The whole methodology was performed in less than two hours with good repeatability of migration times (RSD = 0.91%, n = 5) and corrected peak areas (RSD = 9.6%, n = 5). CE-fingerprints were successfully established for different mAbs and an antibody-drug conjugate.

Simple and Sensitive Method for Deep Profiling of Host Cell Proteins in Therapeutic Antibodies by Combining Ultra-Low Trypsin Concentration Digestion, Long Chromatographic Gradients, and BoxCar Mass Spectrometry Acquisition.

Liquid chromatography coupled to mass spectrometry (LC-MS) is a powerful tool for the analysis of host cell proteins (HCP) during antibody drug process development due to its sensitivity, selectivity, and adaptability. However, the enormous dynamic range between the therapeutic antibody and accompanying HCPs poses a significant challenge for LC-MS based detection of these low abundance impurities. To address this challenge, enrichment of HCPs via immunoaffinity, protein A, 2D-LC, or other strategies is typically performed. However, these enrichments are time-consuming and sometimes require a large quantity of sample. Here, we report a simple and sensitive strategy to analyze HCPs in therapeutic antibody samples without cumbersome enrichment by combining an ultra-low trypsin concentration during digestion under nondenaturing conditions, a long chromatographic gradient, and BoxCar acquisition (ULTLB) on a quadrupole-Orbitrap mass spectrometer. Application of this strategy to the NIST monoclonal antibody standard (NISTmAb) resulted in the identification of 453 mouse HCPs, which is a significant increase in the number of identified HCPs without enrichment compared to previous reports. Known amounts of HCPs were spiked into the purified antibody drug substance, demonstrating that the method sensitivity is as low as 0.5 ppm. Thus, the ULTLB method represents a sensitive and simple platform for deep profiling of HCPs in antibodies.

Morphological and biochemical effects of food deprivation during the early development of Pacific red snapper Lutjanus peru.

We report the effects of food deprivation on the early development of Pacific red snapper Lutjanus peru during the first days of development. The point of no return (PNR) was determined using the feeding incidence after a delay in first feeding. The gradual deterioration of the larvae during food deprivation was recorded using morphometric, histological, enzymatic and biochemical analysis. The time to reach the PNR was 120 h after hatching. Morphologically, the total length, muscle height, head length, tail length and pectoral angle showed the biggest reductions and their growth coefficients changed significantly during food deprivation. Histologically, enterocyte height also was reduced significantly. The protein concentration and activities of the digestive enzymes trypsin, cathepsin-like and lipase showed a significant decrease; meanwhile, amylase activity remained constant during food deprivation. The concentration of total essential free amino acids (EFAAs) decreased significantly while that of the nonessential free amino acids (NEFAAs) remain stable during food deprivation. The most abundant EFAAs were lysine, leucine, isoleucine and valine; the most abundant NEFAAs were alanine, glycine and glutamate, suggesting a more prominent role as energy substrates. At the time of the PNR the concentration of almost all the free amino acids showed a significant decrease. Early food deprivation has a significant impact on the morphology and biochemical characteristics of L. peru. These results suggest that initial feeding of L. peru should begin within 3 days of yolk sac depletion to avoid the PNR. Further studies are necessary to confirm and validate the characters identified in this study as biomarkers of starvation under culture conditions and evaluate their possible utility in ichthyoplankton surveys.

Binding loop of sunflower trypsin inhibitor 1 serves as a design motif for proteolysis-resistant antimicrobial peptides

Although antimicrobial peptides (AMPs) have become powerful drug candidates in the post-antibiotic era, but their low protease stability hinders their clinical application. In the present study, the natural sunflower trypsin inhibitor 1 (SFTI-1) binding loop (CTKSIPPIC) was used to design and synthesize a specific anti-proteolytic sequence template ((RX)n W (RX)n CTKSIPPIC (n = 2, 3; X represents A, I, L, V, F, and W)). After several antibacterial, bactericidal, and toxicity tests, RV3 stood out from the variants and had the highest average selectivity index (SI all = 156.03). It is highly stable in serum, varying pH, temperature, and salt ions as well as under high trypsin, pepsin, or papain concentrations. In a mouse skin inflammation model, established by Pseudomonas aeruginosa infection, RV3 could effectively kill the pathogen, promote wound healing, inhibit inflammatory cell infiltration, and inhibit mRNA and protein expression of TNF-α, IL-6, and IL-1β inflammatory factors. The antibacterial mechanisms of RV3 include combining with lipopolysaccharides and increasing cell membrane permeability, leading to cell membrane rupture and death. These findings indicate that RV3 has great potential for the treatment of bacterial infections.

Peptide modified manganese-doped iron oxide nanoparticles as a sensitive fluorescence nanosensor for non-invasive detection of trypsin activity in vitro and in vivo.

Herein, a fluorescence turn-on nanosensor (MnIO@pep-FITC) has been proposed for detecting trypsin activity in vitro and in vivo through covalently immobilizing an FITC modified peptide substrate of trypsin (pep-FITC) on manganese-doped iron oxide nanoparticle (MnIO NP) surfaces via a polyethylene glycol (PEG) crosslinker. The conjugation of pep-FITC with MnIO NPs results in the quenching of FITC fluorescence. After trypsin cleavage, the FITC moiety is released from the MnIO NP surface, leading to a remarkable recovery of FITC fluorescence signal. Under the optimum experimental conditions, the recovery ratio of FITC fluorescence intensity is linearly dependent on the trypsin concentration in the range of 2 to 100 ng mL−1 in buffer and intracellular trypsin in the lysate of 5 × 102 to 1 × 104 HCT116 cells per mL, respectively. The detection limit of trypsin is 0.6 ng mL−1 in buffer or 359 cells per mL HCT116 cell lysate. The MnIO@pep-FITC is successfully employed to noninvasively monitor trypsin activity in the ultrasmall (ca. 4.9 mm3 in volume) BALB/c nude mouse-bearing HCT116 tumor by in vivo fluorescence imaging with external magnetic field assistance, demonstrating that it has excellent practicability.

The Efficiency of Decellularization of Bovine Pericardium by Different Concentrations of Sodium Dodecyl Sulfate

Background. In modern cardiovasclar surgery, it is a promising method to use xenotissues, which in their properties are close to human tissues, in order to restore the integrity of the heart chambers, its walls or valves. Decellularization of extracellular matrix (EMC) is applied in the process of creation of such bioprotheses. In EMC, the elastin and collagen components are preserved, and antigenic molecules are eliminated resulting in reduction the risk of rejection. The study is devoted to assessment of the histological, molecular-genetic and cytotoxic properties of decellularized bovine pericardium, processed with various concentrations of trypsin enzyme.Objective. The aim of the work is evaluation of efficiency of bovine pericardial decellularization, based on the use of trypsin enzyme with 1% Sodium Dodecyl Sulfate (SDS) and 0.1% SDS.Methods. Bovine pericardium was used as a biomaterial for decellularization. Decellularization protocol 1 envisages processing of samples with 0.25% Trypsin solution at 24 °C with constant shaking (200 rpm) along with processing with 1% ionic SDS detergent. The samples, prepared according to protocol 2, were processed with a low concentration of 0.1% SDS. Histological and morphological properties along with detection of nucleic acids concentration in the samples were studied. Matrix samples were cultured in a human fibroblasts cell culture suspension in order to determine cytotoxicity.Results. Histological examination has not revealed any presence of cells in tissues, decellularized in accordance with both protocols. More than 99% of the nucleic acids was removed from decellularized bovine matrix. During our study, we have not observed cytotoxic effect in vitro for protocol 2 matrix samples, decellularized with only 0.1% SDS. Focal destruction of fibroblasts was observed in conditions of long-term cultivation in protocol 1 samples (Trypsin + 1% SDS). Cells formed abnormal morphological aggregates. Samples of this group have also demonstated sructural changes in collagen and elastin fibers.Conclusions. Studies have shown that pericardial matrix tissue, decellularized with low-concentration of 0.1% SDS, has the same biological properties as the native pericardium. Decellularization of bovine pericardium, using trypsin enzyme with 1% SDS, has a cytotoxic effect on human cells.

Tryptophan Fluorescence and Time-Lag Hydrolysis of Peptide Bonds during Degradation of β-Lactoglobulin by Trypsin

The opening of protein globules and corresponding exposure of their internal peptide bonds, the so-called demasking effect, is required for successful hydrolysis of peptide bonds by proteases. Under the proteolytic action of trypsin on β-lactoglobulin (β-LG), the evolution of tryptophan fluorescence spectra showed that the demasking process consists of two stages with different demasking rate constants for each stage. It was found that the ratio of these constants depends on the concentration of trypsin and changes are approximately threefold when the concentration of trypsin changes in the range of 0.3–15 mg/L. Simulation of hydrolysis taking into account the demasking effect demonstrated how the apparent first-order rate constants obtained experimentally are related to the true hydrolysis rate constants and demasking parameters. The lag phase in the kinetic curves corresponding to the hydrolysis of various peptide bonds in β-LG was also analyzed. The increased lag times indicated sites that are hydrolyzed by a two-stage demasking mechanism.

Insulin Release from NPH Insulin-Loaded Pluronic® F127 Hydrogel in the Presence of Simulated Tissue Enzyme Activity

Background: Despite the widespread use of newer basal insulins, Natural Protamine Hagedorn (NPH) insulin still represents a well-established basal formulation with its long history of use, featuring the native form of human insulin. However, NPH insulin exhibits an undesirable peak within hours after a single subcutaneous (s.c.) injection, which may lead to hypoglycemia followed by insufficient basal insulin delivery. This may be attributed to the s.c. enzyme activities degrading the protamine in NPH microcrystals. Methods: A thermogelling block copolymer Pluronic® F127 (PF127) was utilized as a protective carrier for NPH microcrystals and as a modulator for insulin release from NPH. NPH insulin-loaded PF127 gel was prepared with varying concentrations of the polymer (15–25%) under mild conditions. The formulations were characterized for their gelling temperature, morphology, gel erosion, and in vitro insulin release, with trypsin concentrations up to 5 U/mL. Results: Scanning electron microscopy (SEM) showed that the integrity of NPH microcrystals was maintained after preparation. The burst release of insulin from NPH was significantly attenuated over the course of ~16h in the presence of PF127 with or without enzyme activity. Conclusion: NPH-PF127 successfully resisted the acceleration of NPH crystal dissolution and insulin release in vitro in the presence of protamine-degrading enzyme activity, warranting further testing.

The effect of proteolytic enzymes and pH on GII.4 norovirus, during both interactions and non-interaction with Histo-Blood Group Antigens

Human noroviruses (HuNoVs) are the leading cause of acute gastroenteritis worldwide. Histo-Blood Groups Antigens (HBGAs) have been described as attachment factors, promoting HuNoV infection. However, their role has not yet been elucidated. This study aims to evaluate the ability of HBGAs to protect HuNoVs against various factors naturally found in the human digestive system. The effects of acid pH and proteolytic enzymes (pepsin, trypsin, and chymotrypsin) on GII.4 virus-like particles (VLPs) and GII.4 HuNoVs were studied, both during interactions and non-interaction with HBGAs. The results showed that GII.4 VLPs and GII.4 HuNoVs behaved differently following the treatments. GII.4 VLPs were disrupted at a pH of less than 2.0 and in the presence of proteolytic enzymes (1,500 units/mL pepsin, 100 mg/mL trypsin, and 100 mg/mL chymotrypsin). VLPs were also partially damaged by lower concentrations of trypsin and chymotrypsin (0.1 mg/mL). Conversely, the capsids of GII.4 HuNoVs were not compromised by such treatments, since their genomes were not accessible to RNase. HBGAs were found to offer GII.4 VLPs no protection against an acid pH or proteolytic enzymes.

Generic sensor platform based on electro-responsive molecularly imprinted polymer nanoparticles (e-NanoMIPs)

The present research describes the design of robust electrochemical sensors based on electro-responsive molecularly imprinted polymer nanoparticles (e-MIPs). The e-MIPs, tagged with a redox probe, combine both recognition and reporting functions. This system replaces enzyme-mediator pairs used in traditional biosensors. The analyte recognition process relies on the generic actuation phenomenon when the polymer conformation of e-MIPs is changing in response to the presence of the template analyte. The analyte concentration is measured using voltammetric methods. In an exemplification of this technology, electrochemical sensors were developed for the determination of concentrations of trypsin, glucose, paracetamol, C4-homoserine lactone, and THC. The present technology allows for the possibility of producing generic, inexpensive, and robust disposable sensors for clinical, environmental, and forensic applications.

Influence of ractopamine hydrochloride supplementation on pancreatic digestive enzyme activity in finishing steers

Sixty-nine finishing steers were randomly assigned to one of two treatment groups: supplementation to provide 0 or 267 mg·d−1 of ractopamine hydrochloride (RAC) for 42 d. Twelve steers were slaughtered for tissue and blood collection. Final body weight (BW) tended to be greater with RAC supplementation. Ractopamine hydrochloride supplementation decreased liver mass as a percentage of BW. Pancreatic protein concentration, digestive enzyme activities, serum glucose, and insulin concentrations were not influenced by RAC supplementation. Ractopamine hydrochloride supplementation decreased the pancreatic:α-amylase:trypsin and serum urea nitrogen concentrations. These data suggest that RAC may influence pancreatic exocrine function in cattle.

A Paper-Based Near-Infrared Optical Biosensor for Quantitative Detection of Protease Activity Using Peptide-Encapsulated SWCNTs

A protease is an enzyme that catalyzes proteolysis of proteins into smaller polypeptides or single amino acids. As crucial elements in many biological processes, proteases have been shown to be informative biomarkers for several pathological conditions in humans, animals, and plants. Therefore, fast, reliable, and cost-effective protease biosensors suitable for point-of-care (POC) sensing may aid in diagnostics, treatment, and drug discovery for various diseases. This work presents an affordable and simple paper-based dipstick biosensor that utilizes peptide-encapsulated single-wall carbon nanotubes (SWCNTs) for protease detection. Upon enzymatic digestion of the peptide, a significant drop in the photoluminescence (PL) of the SWCNTs was detected. As the emitted PL is in the near-infrared region, the developed biosensor has a good signal to noise ratio in biological fluids. One of the diseases associated with abnormal protease activity is pancreatitis. In acute pancreatitis, trypsin concentration could reach up to 84 µg/mL in the urine. For proof of concept, we demonstrate the feasibility of the proposed biosensor for the detection of the abnormal levels of trypsin activity in urine samples.

A Preliminary Study on the Ability of the Trypsin-Like Peptidase Activity Assay Kit to Detect Periodontitis.

This study aimed to explore whether the Trypsin-Like Peptidase Activity Assay Kit (TLP-AA-Kit), which measures the activity of N-benzoyl-dl-arginine peptidase (trypsin-like peptidase), can be used as a reliable tool for periodontitis detection in population-based surveillance. In total, 105 individuals underwent a full-mouth periodontal examination and provided tongue swabs as specimens for further analyses. The results of the TLP-AA-Kit were scored between 1 and 5; higher scores indicated higher trypsin concentrations. Receiver operating characteristic analyses were used to evaluate the predictive validity of the TLP-AA-Kit, where the periodontitis case definition provided by the Centers for Disease Control/American Academy of Periodontology served as the reference. Severe and moderate periodontitis were identified in 4.8% and 16.2% of the study population, respectively. The TLP-AA-Kit showed high diagnostic accuracy for severe periodontitis, with an area under the curve of 0.93 (95% confidence interval = 0.88–0.99). However, the diagnostic accuracy of the TLP-AA-Kit for moderate/severe periodontitis was not reliable. While further studies are necessary to validate our results, the results provided herein highlight the potential of the TLP-AA-Kit as a useful tool for the detection of periodontitis, particularly in severe cases, for population-based surveillance.

Assessment of grain and vegetable type soybean genotypes for nutritional composition

The present study was conducted to analyse the nutritional composition of fourteen soybean genotypes including seven each of grain and vegetable types. The study revealed that oil content was in the range of 17.51 to 20.59% at R8 stage, protein 18.11 % to 27.57% at R6 and 32.57 % to 39.09% at R8 stage, carbohydrate content at R6 stage 7.43 to 11.08 g/100g and 24.15 to 28.37 g/100g at R8 stage, sugar content 0.90 to 3.43 g/100g, iron content at R6 stage 3.24 to 7.12 mg/100g and 6.79 mg/100g to 9.79 mg/100g at R8 stage, zinc content at R6 stage 4.74 to 6.40 mg/100g and 3.93 mg/100g to 5.60 mg/100 g, The trypsin inhibitor content was ranged between 18.29 to 46.26 mg/g and 33.96 to 75.09 mg/g at R6 and R8 stage respectively. Vegetable types were found more nutritious than grain type soybean. Swarna vasundhara was found promising for protein content at R6 stage, the genotype AGS-450 exhibited high iron with low trypsin content whereas, AGS-459 exhibited high sugar and zinc content.