Allosteric Inhibition of Factor XIIIa. Non-Saccharide Glycosaminoglycan Mimetics, but Not Glycosaminoglycans, Exhibit Promising Inhibition Profile

Factor XIIIa (FXIIIa) is a transglutaminase that catalyzes the last step in the coagulation process. Orthostery is the only approach that has been exploited to design FXIIIa inhibitors. Yet, allosteric inhibition of FXIIIa is a paradigm that may offer a key advantage of controlled inhibition over orthosteric inhibition. Such an approach is likely to lead to novel FXIIIa inhibitors that do not carry bleeding risks. We reasoned that targeting a collection of basic amino acid residues distant from FXIIIa’s active site by using sulfated glycosaminoglycans (GAGs) or non-saccharide GAG mimetics (NSGMs) would lead to the discovery of the first allosteric FXIIIa inhibitors. We tested a library of 22 variably sulfated GAGs and NSGMs against human FXIIIa to discover promising hits. Interestingly, although some GAGs bound to FXIIIa better than NSGMs, no GAG displayed any inhibition. An undecasulfated quercetin analog was found to inhibit FXIIIa with reasonable potency (efficacy of 98%). Michaelis-Menten kinetic studies revealed an allosteric mechanism of inhibition. Fluorescence studies confirmed close correspondence between binding affinity and inhibition potency, as expected for an allosteric process. The inhibitor was reversible and at least 9-fold- and 26-fold selective over two GAG-binding proteins factor Xa (efficacy of 71%) and thrombin, respectively, and at least 27-fold selective over a cysteine protease papain. The inhibitor also inhibited the FXIIIa-mediated polymerization of fibrin in vitro. Overall, our work presents the proof-of-principle that FXIIIa can be allosterically modulated by sulfated non-saccharide agents much smaller than GAGs, which should enable the design of selective and safe anticoagulants

Plasmin Regulation through Allosteric, Sulfated, Small Molecules

Plasmin, a key serine protease, plays a major role in clot lysis and extracellular matrix remodeling. Heparin, a natural polydisperse sulfated glycosaminoglycan, is known to allosterically modulate plasmin activity. No small allosteric inhibitor of plasmin has been discovered to date. We screened an in-house library of 55 sulfated, small glycosaminoglycan mimetics based on nine distinct scaffolds and varying number and positions of sulfate groups to discover several promising hits. Of these, a pentasulfated flavonoid-quinazolinone dimer 32 was found to be the most potent sulfated small inhibitor of plasmin (IC50 = 45 μM, efficacy = 100%). Michaelis-Menten kinetic studies revealed an allosteric inhibition of plasmin by these inhibitors. Studies also indicated that the most potent inhibitors are selective for plasmin over thrombin and factor Xa, two serine proteases in coagulation cascade. Interestingly, different inhibitors exhibited different levels of efficacy (40%–100%), an observation alluding to the unique advantage offered by an allosteric process. Overall, our work presents the first small, synthetic allosteric plasmin inhibitors for further rational design

Reef sponges facilitate the transfer of coral-derived organic matter to their associated fauna via the sponge loop

The high biodiversity of coral reefs results in complex trophic webs where energy and nutrients are transferred between species through a multitude of pathways. Here, we hypothesize that reef sponges convert the dissolved organic matter released by benthic primary producers (e.g. corals) into particulate detritus that is transferred to sponge-associated detritivores via the sponge loop pathway. To test this hypothesis, we conducted stable isotope (13C and15N) tracer experiments to investigate the uptake and transfer of coral-derived organic matter from the sponges Mycale fistulifera and Negombata magnifica to 2 types of detritivores commonly associated with sponges: ophiuroids (Ophiothrix savignyi and Ophiocoma scolopendrina) and polychaetes (Polydorella smurovi). Findings revealed that the organic matter naturally released by the corals was indeed readily assimilated by both sponges and rapidly released again as sponge detritus. This detritus was subsequently consumed by the detritivores, demonstrating transfer of coral-derived organic matter from sponges to their associated fauna and confirming all steps of the sponge loop. Thus, sponges provide a trophic link between corals and higher trophic levels, thereby acting as key players within reef food webs

Influence of cupping treatment on high-intensity anaerobic performance

The use of cupping therapy prior to sports events has increased in popularity, with limited evidence to support its efficacy. The purpose of this study was to evaluate the efficacy of dry and wet cupping therapy on subsequent Wingate anaerobic test (WAnT) performance. Twelve trained men participated in this repeated measures randomized crossover study (age 24.9 ± 4.8 years; body mass index 27.6 ± 14.3 kg.m-2). Participants were familiarized with the ergometer and the Wingate anaerobic test on three separate occasions. They then randomly performed three experimental Wingate tests separated by 48-72 h after either dry cupping (DRY), wet cupping (WET), or no treatment (CON). Repeated measures ANOVA and Pearson’s correlation coefficient were used to analyze data and determine the relationships between WAnT and peak lactate and heart rate (HR). Peak power (PP), mean power (MP), and fatigue index (FI) were similar in all treatments (p=.47-.72). Heart rate (HR) and lactate increased similarly at all time points in all treatments (p<.001 for all comparisons). Post-WAnT peak HR was moderately negatively correlated with PP in all treatments and MP in CON only (p<.05 for all correlations). No other significant correlations were detected. The present findings demonstrate no beneficial effects of wet and dry cupping therapy, and hence do not support its use prior to high-intensity anaerobic sports events

The Discrete Fractional Variable-Order Tinkerbell Map: Chaos, 0–1 Test, and Entropy

The dynamics of the Caputo-fractional variable-order difference form of the Tinkerbell map are studied. The phase portraits, bifurcation, and largest Lyapunov exponent (LLE) were employed to demonstrate the presence of chaos over a different fractional variable-order and establish the nature of the dynamics. In addition, the 0-1 test tool was used to detect chaos. Finally, the numerical results were confirmed using the approximate entropy

Isolation, characterization, and hydrolytic activities of Geobacillus species from Jordanian hot springs

The present study was conducted to isolate, identify, characterize and to determine the enzymatic activities of the thermophilic Geobacillus species from five Jordanian hot springs. Based on phenotypic characters, eight thermophilic isolates were identified and belonged to the genus Geobacillus. The Geobacillus isolates were abundant in all investigated hot springs. The optimal temperature for growth of the isolates was 60 to 65°C and the optimal pH was 6 to 8. Colonies were light yellow circular to rhizoid. The bacterial cells were Gram positive rods and endospore forming. All isolates produced amylase, caseinase, alkaline and acid phosphatases, esterase (C4), esterase lipase (C8), α-Galactosidase, β-Glucuronidase, β-Glucosidase, and N-Acetyl-β-glucosaminidase. Seven isolates produced leucine and valine arylamidases and five isolates produced naphthol-AS-B1- phsphohydrolase. Lipase (C14) activity from two isolates and α-chymotrypsin activity from three isolates were also detected. The phenotypic characterization of those isolates was confirmed by genotypic method using 16S rDNA sequence analysis. Maximal homology of all eight  isolates to genus Geobacillus was observed. Five of these isolates showed greater than 98% homology with Geobacillus stearothermophilus and one isolate showed 100% homology with Geobacillus thermoglucosidasius. Therefore, 16S rRNA gene sequence analysis can be considered as a valuable genotypic tool for the identification and characterization of thermophilic bacteria at genus level. Moreover, enzymatic products of those isolates could receive considerable attention due to their potential applications in biotechnology.Keywords: Thermophiles, Geobacillus, hydrolytic enzymes, hot spring, 16S rRNA

Anemonefish oxygenate their anemone hosts at night

Many stony coral-dwelling fishes exhibit adaptations to deal with hypoxia among the branches of their hosts; however, no information exists on the respiratory ecophysiology of obligate fish associates of non-coral organisms such as sea anemones and sponges. This study investigated metabolic and behavioral interactions between two-band anemonefish (Amphiprion bicinctus) and bulb-tentacle sea anemones (Entacmaea quadricolor) at night. We measured the net dark oxygen uptake (VO2, μmol O2h–1) of fish–anemone pairs when partners were separate from each other, together as a unit, and together as a unit but separated by a mesh screen that prevented physical contact. We also measured the effects of water current on sea anemone VO2 and quantified the nocturnal behaviors of fish in the absence and presence of host anemones in order to discern the impacts of anemone presence on fish behavior. Net VO2 of united pairs was significantly higher than that of both separated pairs and united pairs that were separated by a mesh screen. Anemone VO2 increased with flow rate from 0.5 to 2.0cms–1, after which VO2 remained constant up to a water flow rate of 8.0cms–1. Furthermore, the percentage time and bout frequency of flow-modulating behaviors by fish increased significantly when anemones were present. We conclude that physical contact between anemonefish and sea anemones elevates the VO2 of at least one of the partners at night, and anemonefish behavior at night appears to oxygenate sea anemone hosts and to augment the metabolism of both partners.PublishedYe

The new fractional discrete neural network model under electromagnetic radiation: Chaos, control and synchronization

This paper describes a new four-dimensional fractional discrete neural network with electro-magnetic radiation model. In addition, the non-linear dynamics of the suggested model are examined, within the framework of commensurate, incommensurate and variable orders, through different numer-ical techniques such as Lyapunov exponent, phase portraits, bifurcation diagrams, and the 0-1 test method. The results imply that The behaviours of the proposed fractional discrete neural network model have rich and complex dynamical properties that are influenced by the variation of the system parame-ters, the commensurate order, the incommensurate order and the variable order. Moreover, the approx-imate entropy test and Co algorithm are carried out to measure complexity and validate the presence of chaos. Finally, nonlinear controllers are illustrated to stabilize and synchronize the proposed model. & COPY; 2023 THE AUTHORS. Published by Elsevier BV on behalf of Faculty of Engineering, Alexandria University. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/ licenses/by-nc-nd/4.0/)

Effect of Various Local Anthropogenic Impacts on the Diversity of Coral Mucus-Associated Bacterial Communities

The global continued decline in coral reefs is intensifying the need to understand the response of corals to local environmental stressors. Coral-associated bacterial communities have been suggested to have a swift response to environmental pollutants. This study aims to determine the variation in the bacterial communities associated with the mucus of two coral species, Pocillopora damicornis (Linnaeus, 1758) and Stylophora pistillata (Esper, 1792), and the coral-surrounding seawater from three areas exposed to contamination at the Jordanian coast of the Gulf of Aqaba (Red Sea), and also explores the antibacterial activity of these bacteria. Corals were collected from three contaminated zones along the coast, and the bacteria were quantified and identified by conventional morphological and biochemical tests, as well as 16S rRNA gene sequencing. The average number of bacteria significantly varied among the coral mucus from the sampling zones and between the coral mucus and the surrounding seawater. The P. damicornis mucus-associated bacterial community was dominated by members of the classes Gammaproteobacteria, Cytophagia, and Actinomycetia, while the mucus of S. pistillata represented higher bacterial diversity, with the dominance of the bacterial classes Gammaproteobacteria, Actinomycetia, Alphaproteobacteria, and Bacilli. The effects of local anthropogenic impacts on coral mucus bacterial communities were represented in the increased abundance of bacterial species related to coral diseases. Furthermore, the results demonstrated the existence of bacterial isolates with antibacterial activity that possibly acted as a first line of defense to protect and maintain the coral host against pathogens. Indeed, the dynamics of coral-associated microbial communities highlight the importance of holistic studies that focus on microbial interactions across the coral reef ecosystem

Past 140-year environmental record in the northern South China Sea: Evidence from coral skeletal trace metal variations

About 140-year changes in the trace metals in Porites coral samples from two locations in the northern South China Sea were investigated. Results of PCA analyses suggest that near the coast, terrestrial input impacted behavior of trace metals by 28.4%, impact of Sea Surface Temperature (SST) was 19.0%, contribution of war and infrastructure were 14.4% and 15.6% respectively. But for a location in the open sea, contribution of War and SST reached 33.2% and 16.5%, while activities of infrastructure and guano exploration reached 13.2% and 14.7%. While the spatiotemporal change model of Cu, Cd and Pb in seawater of the north area of South China Sea during 1986-1997 were reconstructed. It was found that in the sea area Cu and Cd contaminations were distributed near the coast while areas around Sanya, Hainan had high Pb levels because of the well-developed tourism related activities. (C) 2013 Elsevier Ltd. All rights reserved