Current knowledge on the extraction,purification, identification,and validation of bioactive peptides from seaweed

Seaweed (macroalgae) is considered as a sustainable bioresource rich in high-quality nutrients such as protein. Seaweed protein can be used as an alternative to other protein sources. Furthermore, these proteins are natural reservoirs of bioactive peptides (BAPs) associated with various health benefits such as antioxidant, antihypertensive, and antidiabetic activities. However, seaweed-derived BAPs remain underexploited due to challenges that arise during protein extraction from algal biomass. Coupled with this, limited proteomic information exists regarding certain seaweed species. This review highlights the current state of the art of seaweed protein extraction techniques, e.g., liquid, ultrasound, microwave, pulsed electric field, and high hydrostatic pressure assisted extraction. The review also focuses on the enzymatic hydrolysis of seaweed proteins and characterization of the resultant hydrolysates/peptides using electrophoretic and chromatographic techniques. This includes reference to methods employed for separation, fractionation, and purification of seaweed BAPs, as well as the methodologies used for identification, e.g., analysis by mass spectrometry. Furthermore, a bioinformatics or in silico approach to aid discovery of seaweed BAPs is discussed herein. Based on the information available to date, it is suggested that further research is required in this area for the development of seaweed BAPs for nutraceutical applications.     

Tooth-root stress calculation of high transverse contact ratio spur and helical gears

In this paper, a non-uniform model of load distribution along the line of contact of spur and helical gears, obtained from the minimum elastic potential criterion, has been used, combined with the equations of the linear elasticity, to evaluate the tooth-root stress of high transverse contact ratio gears. The values of both critical stress and load conditions have been obtained and a complete analysis of the tooth bending strength has been carried out. As the load per unit of length at any point of the line of contact and any position of the meshing cycle has been described by a very simple equation, a complete study of the location and the value of the tooth-root stress has been carried out. From this study, a recommendation for the calculation of the bending load capacity of high transverse contact ratio spur and helical gears is proposed.     

Rapid and scalable synthesis of chiral bromolactones as precursors to α-exo-methylene-γ-butyrolactone-containing sesquiterpene lactones

The sesquiterpene lactones cover a diverse and pharmacologically important diversity space. In particular, the electrophilic α-exo-methylene-γ-butyrolactone moiety that is preponderant in this natural product family has been shown to readily engage in covalent inhibition via conjugate addition of cysteine residues in target proteins. However, the synthetic accessibility of sesquiterpenes or related probes to investigate their mode of action remains laborious. Herein, we present a rapid and scalable route to chiral bromolactones as enabling precursors in the synthesis of sesquiterpene lactones.     

Synthesis of carbamoyl azides from primary amines and carbon dioxide under mild conditions

Treatment of amines under a carbon dioxide atmosphere with tetramethylphenylguanidine (PhTMG) and diphenylphosphoryl azide (DPPA) in acetonitrile below 0 degrees C provides carbamoyl azides in high to excellent yields. In addition, epimerization is not observed when optically pure alpha-amino esters are used as substrates.     

Enhanced model of load distribution along the line of contact for non-standard involute external gears

The presence of undercut at the tooth root, non-equal addendum on pinion and wheel, non-standard tooth height or non-standard center distance may have decisive influence on the load distribution along the line of contact of spur and helical gear teeth. The curve of variation of the meshing stiffness along the path of contact, quite symmetric respect the midpoint of the interval of contact, loses its symmetry for non-standard geometries and operating conditions. As a consequence, the critical contact points for bending and wear calculations may be shifted from their locations for standard gears. In this paper, a non-uniform model of load distribution along the line of contact of standard spur and helical gears, obtained from the minimum elastic potential criterion, has been enhanced to fit with the meshing conditions of the above mentioned non-standard cylindrical gear pairs. The same analytical formulation of the initial model may be used for the non-standard gears by considering appropriate values of a virtual contact ratio, which are also presented in the paper.     

How not to formulate multiple choice problems

This note discusses some of the shortcomings of multiple choice tests in Mathematics given to undergraduate engineering students. Examples are presented, where the disadvantages of a multiple choice test are pointed out and suggestions how to overcome the difficulties are given.     

Decomposing Timed Petri Net Models of Open Queueing Networks

This paper develops a Petri net-based decomposition approach to model and perform steady-state analysis of open queueing networks with multi-class customers. For each customer class we assume deterministic routeing and different service time distribution. The current decomposition may also consider equipment failures, defective units that have to be re-worked and on-line preparations (to switch from one mix of customers to another) as additional customer classes. In contrast to most of the published works on timed Petri nets, their present use is marked by input flows, originating from external sources. A simple method, directly derived from the timed Petri nets theories, is used for calculating the expected utilization of each server and eventually the maximal productivity of the system for the given deterministic routeing. An important application of the model is in validating discrete-event non-terminating simulation models.     

Quantitative and confirmative performance of liquid chromatography coupled to high-resolution mass spectrometry compared to tandem mass spectrometry

The quantitative and confirmative performance of two different mass spectrometry (MS) techniques (high-resolution MS and tandem MS) was critically compared. Evaluated was a new extraction and clean-up protocol which was developed to cover more than 100 different veterinary drugs at trace levels in a number of animal tissues and honey matrices. Both detection techniques, high-resolution mass spectrometry (HRMS) (single-stage Orbitrap instrument operated at 50 000 full width at half maximum) and tandem mass spectrometry (MS/MS) (quadrupole technology) were used to validate the method according to the EU Commission Decision 2002/657/EEC. Equal or even a slightly better quantitative performance was observed for the HRMS-based approach. Sensitivity is higher for unit mass resolution MS/MS if only a subset of the 100 compounds has to be monitored. Confirmation of suspected positive findings can be done by evaluating the intensity ratio between different MS/MS transitions, or by accurate mass based product ion traces (no precursor selection applied). MS/MS relies on compound-specific optimized transitions; hence the second, confirmatory transition generally shows relatively high ion abundance (fragmentation efficacy). This is often not the case in single-stage HRMS, since a generic (not compound-optimized) collision energy is applied. Hence, confirmation of analytes present at low levels is superior when performed by MS/MS. Slightly better precision, but poorer accuracy (fortified matrix extracts versus pure standard solution) of ion ratios were observed when comparing data obtained by HRMS versus MS/MS.     

Supported Catalytically Active Supramolecular Hydrogels for Continuous Flow Chemistry

Inspired by biology, one current goal in supramolecular chemistry is to control the emergence of new functionalities arising from the self‐assembly of molecules. In particular, some peptides can self‐assemble and generate exceptionally catalytically active fibrous networks able to underpin hydrogels. Unfortunately, the mechanical fragility of these materials is incompatible with process developments, relaying this exciting field to academic curiosity. Here, we show that this drawback can be circumvented by enzyme‐assisted self‐assembly of peptides initiated at the walls of a supporting porous material. We applied this strategy to grow an esterase‐like catalytically active supramolecular hydrogel (CASH) in an open‐cell polymer foam, filling the whole interior space. Our supported CASH material is highly efficient towards inactivated esters and enables the kinetic resolution of racemates. This hybrid material is robust enough to be used in continuous flow reactors, and is reusable and stable over months.