α-Glucosidase (AG) is a bifunctional enzyme, it has a capacity to synthesize 2-O-α-D-glucopyranosyl-L-ascorbic acid (AA-2G) from L-ascorbic acid (L-AA) and low-cost maltose under mild conditions, but it can also hydrolyze AA-2G, which leads to low synthesis efficiency of AA-2G. Main Methods and Major Results This study introduces a rational molecular design strategy to regulate enzymatic reactions based on inhibiting the formation of ground state of enzyme-substrate complex. Y215 was analyzed as the key amino acid site affecting the affinity of AG to AA-2G and L-AA. For the purpose of reducing the hydrolysis efficiency of AA-2G, the mutant Y215W was obtained by analyzing the molecular docking binding energy and hydrogen bond formation between AG and the substrates. Compared with the wild type, Isothermal Titration Calorimetry(ITC) results showed that the equilibrium dissociation constant (KD) of the mutant for AA-2G was doubled; the Michaelis constant (Km) for AA-2G was reduced by 1.15 times; and the yield of synthetic AA-2G was increased by 39%. Conclusions and Implications Our work also provides a new reference strategy for the molecular modification of multifunctional enzymes and other enzymes in cascade reactions system.

Biocatalysis in high-concentration organic solvents has been applied to produce various industrial products with many advantages. However, using enzymes in organic solvents often suffers from inactivation or decreased catalytic activity and stability. So, improving the tolerance of enzymes in organic solvents is essential. Herein, the method of regional random mutation combined with combinatorial mutation was used to improve the resistance of transaminase from Aspergillus terreus (AtATA) in organic solvents, and the best mutant T23I/T200K/P260S (M3) was acquired. In different concentrations of dimethyl sulfoxide (DMSO), the catalytic efficiency toward 1-acetylnaphthalene and the stability were higher than the wild-type (WT) of AtATA. M3 also showed enhanced stability against six organic solvents with different oil-water partition coefficients (log P values). The results of decreased Root Mean Square Fluctuation (RMSF) values via 20-ns molecular dynamics simulations under different concentration DMSO revealed that mutant M3 had lower flexibility, acquiring a more stable protein structure and contributing to its organic solvents stability than WT. Intra- and intermolecular interaction analysis indicated that the increased hydrogen bonds and hydrophobic interactions within monomers or at the interface of two monomers also strengthened the stability of the overall structure against organic solvents. Furthermore, M3 was applied to convert 1-acetylnaphthalene for synthesizing (R)-(+)-1(1-naphthyl)-ethylamine ((R)-NEA), which was and an intermediate of Cinacalcet Hydrochloride. Moreover, 3~10 mM 1-acetylnaphthalene can be converted to (R)-NEA with 94.2~38.9% yield and a strict R-stereoselectivity within 10 h under 25% DMSO, which was higher than WT and expected to be a potential biocatalyst for industrial application.

TITLEA gigantic iliopsoas abscess in a patient with Alexander’s disease

This study's primary objective is to demonstrate how diffusion-thermo and thermal diffusion influence of peristaltic flow processes with slip boundaries when joule heating happens from the interior. Several operational factors and their impacts on the system were analysed, along with the corresponding graphs. As slip parameters rise, the axial pressure gradient fluid flow tends to drop. The pressure rate is demonstrated to drop in the backward and peristaltic pumping regions as the quantity of the second order slide parameter rises, whereas it rises in the co-pump zone. As slip parameters rise, fluid temperature and concentration tend to drop. Changes in the thermal diffusion and thermo diffusion factors cause changes in the fluid's temperature and concentration. The Nusselts number can be increased by increasing the Prandtl number, the thermo-diffusion constraint, the dufour number, and the Schmidt number. However, this will result in fewer Sherwood number.

Aim Dysfunction of nitric oxide (NO) – soluble guanylate cyclase (sGC) – cyclic guanosine monophosphate (cGMP) signalling is implicated in the pathophysiology of cognitive impairment and dementia. Zagociguat is a central nervous system-(CNS-) penetrant sGC stimulator designed to amplify NO-cGMP signalling in the CNS. This article reports on a phase 1b study evaluating the safety and pharmacodynamic effects of zagociguat. Methods In this randomized crossover study, 24 healthy participants ≥65 years of age were planned to receive 15 mg zagociguat or placebo once daily for two 15-day periods separated by a 27-day washout. Adverse events, vital signs, electrocardiograms, and laboratory tests to assess safety. Pharmacokinetics of zagociguat were evaluated in blood and CSF. Pharmacodynamic assessments included evaluation of cerebral blood flow, CNS tests, pharmaco-electroencephalography, passive leg movement, and biomarkers in blood, cerebrospinal fluid, and brain. Results Twenty-four participants were enrolled and 12 participants completed both treatment periods, while 12 participants completed only one treatment period. Zagociguat was well tolerated and penetrated the blood-brain barrier. Zagociguat induced modest decreases in blood pressure. No consistent effects of zagociguat on other pharmacodynamic parameters were detected. Conclusion Zagociguat was well tolerated and induced modest systemic blood pressure reductions consistent with other sGC stimulators. No clear pharmacodynamic effects of zagociguat were detected, perhaps due to optimal CNS function in healthy participants. Studies in participants with proven reduced cerebral blood flow or CNS function may be an avenue for further evaluation of the compound.

Recently, superparamagnetic silica encapsulated DNA microparticles (SiDNAFe) were designed and in various experiments used as a hydrological tracer. We investigated the effect of bed characteristics on the transport behaviour and especially the mass loss of SiDNAFe in open channel injection experiments. Hereto, a series of laboratory injection experiments were conducted with four channel bed conditions (no sediment, fine river sediment, coarse sand, and goethite-coated coarse sand) and two water qualities (tap water and Meuse water). Breakthrough curves (BTCs) were analysed and modelled. Mass loss of SiDNAFe was accounted for as a first-order decay process included in a 1-D advection and dispersion model with transient storage (OTIS). SiDNAFe BTCs could be adequately described by advection and dispersion with or without a first-order decay process. Mass loss of SiDNAFe increased as a function of the surface roughness of the beds. Retention of SiDNAFe due to surface roughness was 1-2 orders of magnitude greater than gravitational settling rates, as determined in Tang et al. (2022). We speculate this was due to boundary layer kinetic attachment. The dispersive behaviour of SiDNAFe generally mimicked that of NaCl tracer, although SiDNAFe traveled faster on average due to a smaller effective cross-sectional area. No pattern was observed between SiDNAFe mass recovery and water qualities used. DNA concentration data uncertainty was mostly associated with lower SiDNAFe concentrations in the BTCs. This research highlights that riverbeds are important sinks, and the surface roughness affects the fate and transport characteristics of SiDNAFe when in proximity to the water-sediment interface. SiDNAFe possess promising potential as a surrogate for multi-tracing micro-contaminants (e.g., microplastics) in large rivers, which could be a promising tool for enhancing understanding of hydrological processes.

Hepatocellular carcinoma has been a serious threat to human life and health, and there is an urgent need for new treatments to prolong the overall survival time of patients. The liver plays an immunomodulatory function due to its unique physiological structural characteristics; therefore, following surgical resection and radiotherapy, immunotherapeutic options have shown great potential in the treatment of hepatocellular carcinoma in recent years, and adoptive cellular immunotherapy is developing rapidly in the treatment of hepatocellular carcinoma. In this review we summarize the latest research on adoptive cell therapy for hepatocellular carcinoma, focusing on chimeric antigen receptor (CAR) T cells and T cell receptor-engineered (TCR) T cells, and then briefly discuss tumor infiltrating lymphocytes (TILs), natural killer (NK) cells and cytokine-induced killer cells (CIKs). The aim is to provide readers with a comprehensive understanding of the current status of HCC adoptive cellular immunotherapy and new therapeutic strategies being developed, in the hope of providing new ideas for the clinical management of hepatocellular carcinoma.

Fear overgeneralization and perceived uncertainty about future outcomes have been suggested as risk factors for clinical anxiety. However, little is known regarding how they influence each other. In this study, we investigated whether different levels of threat uncertainty influence fear generalization. Three groups of healthy participants underwent a differential fear conditioning protocol followed by a generalization test. All groups learned to associate one female face (conditioned stimulus, CS+) with a female scream (unconditioned stimulus, US) while the other face (CS-) was not associated with the scream. In order to manipulate threat uncertainty, one group (low uncertainty, n = 26) received 80%, the second group (moderate uncertainty, n = 32) received 60%, and the third group (high uncertainty, n = 30) 40% CS-US contingency. In the generalization test, all groups saw CS+ and CS- again as well as four morphs that varied in similarity with the CS+ in steps of 20%. Subjective (expectancy, valence, and arousal ratings), psychophysiological (skin conductance response, SCR), and visuocortical (steady-state visual evoked potentials, ssVEPs) indices of fear were registered. Participants expected the US in accordance with their reinforcement schedules but displayed stronger skin conductance with more uncertainty. However, acquisition of conditioned fear was not evident in ssVEPs. During the generalization test, we found no effect of threat uncertainty in any of the measured variables, but the strength of generalization for threat expectancy ratings was positively correlated with dispositional intolerance of uncertainty. This study suggests that mere threat uncertainty does not modulate fear generalization.

Intestinal helminth infection promotes a Type 2 inflammatory response in resistant C57BL/6 mice that is essential for worm clearance. The study of inbred mouse strains has revealed factors that are critical for parasite resistance and delineated the role of Type 1 versus Type 2 immune responses in worm clearance. In C57BL/6 mice, basophils are key innate immune cells that promote Type 2 inflammation and are programmed via the Notch signaling pathway during infection with the helminth Trichuris muris. However, how the host genetic background influences basophil responses and basophil expression of Notch receptors remains unclear. Here we use genetically susceptible inbred AKR/J mice that have a Type 1-skewed immune response during T. muris infection to investigate basophil responses in a susceptible host. Basophil population expansion occurred in AKR/J mice even in the absence of fulminant Type 2 inflammation during T. muris infection. However, basophils in AKR/J mice did not robustly upregulate expression of the Notch2 receptor in response to infection as in C57BL/6 mice. Blockade of the Type 1 cytokine IFN-γ in infected AKR/J mice was not sufficient to elicit infection-induced basophil expression of the Notch2 receptor. These data suggest that the host genetic background, outside of the Type 1 skew, is important in regulating basophil responses during T. muris infection in susceptible AKR/J mice.

A 16-year-old male with past medical history of congenital atrial septal defect surgical repair presented with recurrent pericarditis secondary to post-cardiotomy injury syndrome (PCIS). After failing medical therapy, he ultimately underwent pericardiectomy for symptom resolution. PCIS is underdiagnosed in children and should be considered in patients with recurrent chest pain.

Aim:This study aims to investigate the effects of maternal asthma on fetal cardiac functions. Methods:The study was planned with 30 pregnant women who presented to a tertiary health center and were diagnosed with asthma and 60 healthy controls with similar gestational ages. The fetal echocardiographic assessment was assessed between 33 and 35 weeks of gestation with pulsed-wave Doppler (PW), M-mode, and tissue Doppler imaging (TDI). Fetal cardiac functions were compared between maternal asthma and control group. Cardiac functions were assessed according to the duration of maternal asthma diagnosis, as well. Results:Early diastolic function parameters, tricuspid E wave (p=0.001), and tricuspid E/A ratio (P=0.005) were significantly lower in the group with maternal asthma. TAPSE and MAPSE values were statistically lower in the study group than in the control group; p=0.010 and p=0.012, respectively. Parameters assessed with TDI (E’, A’, S’, E/E’, and MPI’ of tricuspid valves) and global cardiac function parameters assessed with pulsed-wave Doppler like myocardial performance index (MPI) and left cardiac output (LCO) were similar between groups (p> 0.05). Although, MPI did not change between groups, and the isovolumetric relaxation time (IVRT) value was prolonged in maternal asthma cases (p=0.025). Conclusion:We found that maternal asthma disease causes alteration in fetal diastolic and early systolic cardiac functions, but the global fetal cardiac function does not change. Diastolic heart function values also varied with the duration of maternal asthma. Prospective studies are needed to compare fetal cardiac functions with additional patient groups according to disease severity and type of medical treatment.

Inter-specific adoption is an intriguing topic in behavioural and evolutionary ecology. Being a rare phenomenon is rarely documented in the literature and thus reports of inter-specific adoption based on solid data are particularly valuable. Here, owing to a long-term and extensive monitoring of a local population of the European blackbird (Turdus merula, hereafter blackbird), we describe observations of alloparental behaviour exhibited by blackbirds towards fieldfare (Turdus pilaris) nestlings (single nest, first record ever) and fledglings (twelve cases in total). We discuss the observations in respect to available literature.

Objective: Device lead-induced tricuspid regurgitation (LITR) mechanisms are well-defined by 3D transthoracic echocardiography (3D-TTE). There is a lack of data on the Latin-American population. The objective of this study was to describe the prevalence of several mechanisms and insights in patients with permanent right ventricular (RV) implanted devices by 3D-TTE examination. Methods: We performed a cross-sectional analysis of 101 patients with permanent cardiac devices such as pacemakers or defibrillators. 3D-TTE was obtained on all patients in RV-focused apical views to perform a complete tricuspid valve (TV) evaluation: leaflets, subvalvular apparatus, precise lead location, and functional assessment to evaluate possible mechanisms of tricuspid regurgitation (TR). Results: In a total of 101 patients, the leads did not interfere with TV function in 53 p. (59%), while LITR was observed in 38 (41%) patients. Adherent, impinging, entangled, and mixed lead-induced mechanisms were observed. Time in years since device implantation was significantly higher in patients with LITR. Conclusions: LITR was present in a high proportion of our population. LITR is the result of damage to the TV as well as its subvalvular apparatus due to the fibrotic and inflammatory response over time when leads are situated in unfavorable locations.

Native mass spectrometry is a rapidly emerging technique for fast and sensitive structural analysis of protein constructs, maintaining the protein higher order structure. The coupling with electromigrative separation techniques under native conditions enables the characterization of proteoforms and highly complex protein mixtures. In this review, we present an overview of current native CE-MS technology. First, the status of native separation conditions is described for capillary zone electrophoresis (CZE), affinity capillary electrophoresis (ACE), and capillary isoelectric focusing (CIEF), as well as their chip-based formats, including essential parameters such as electrolyte composition and capillary coatings. Further, conditions required for native ESI-MS of (large) protein constructs, including instrumental parameters of QTOF and Orbitrap systems, as well as requirements for native CE-MS interfacing are presented. On this basis, methods and applications of the different modes of native CE-MS are summarized and discussed in the context of biological, medical, and biopharmaceutical questions. Finally, key achievements are highlighted and concluded, while remaining challenges are pointed out.

Estimating the accuracy of quaternary structural models of protein complexes and assemblies (EMA) is important for predicting quaternary structures and applying them to studying protein function and interaction. The pairwise similarity between structural models is proven useful for estimating the quality of protein tertiary structural models, but it has been rarely applied to predicting the quality of quaternary structural models. Moreover, the pairwise similarity approach often fails when many structural models are of low quality and similar to each other. To address the gap, we developed a hybrid method (MULTICOM_qa) combining a pairwise similarity score (PSS) and an interface contact probability score (ICPS) based on the deep learning inter-chain contact prediction for estimating protein complex model accuracy. It blindly participated in the 15th Critical Assessment of Techniques for Protein Structure Prediction (CASP15) in 2022 and ranked first out of 24 predictors in estimating the global accuracy of assembly models. The average per-target correlation coefficient between the model quality scores predicted by MULTICOM_qa and the true quality scores of the models of CASP15 assembly targets is 0.66. The average per-target ranking loss in using the predicted quality scores to rank the models is 0.14. It was able to select good models for most targets. Moreover, several key factors (i.e., target difficulty, model sampling difficulty, skewness of model quality, and similarity between good/bad models) for EMA are identified and analayzed. The results demonstrate that combining the multi-model method (PSS) with the complementary single-model method (ICPS) is a promising approach to EMA.

Processing of CASP15 targets into evaluation units (EUs) and assigning them to evolutionary-based prediction classes is presented in this study. The targets were first split into structural domains based on compactness and similarity to other proteins. Models were then evaluated against these domains and their combinations. The domains were joined into larger EUs if predictors’ performance on the combined units was similar to that on individual domains. Alternatively, if most predictors performed better on the individual domains, then they were retained as EUs. As a result, 112 evaluation units were created from 77 tertiary structure prediction targets. The EUs were assigned to four prediction classes roughly corresponding to target difficulty categories in previous CASPs: TBM (template-based modeling, easy or hard), FM (free modeling), and the TBM/FM overlap category. More than a third of CASP15 EUs were attributed to the historically most challenging FM class, where homology or structural analogy to proteins of known fold cannot be detected.

A document by Eduardo Howard. Click on the document to view its contents.

Background: For severe mitral valve (MV) degenerative disease, repair is recommended. Prediction of repair complexity and referral to centers of excellence can increase rates of successful repair. This study sought to demonstrate that TEE is a feasible imaging modality to predict the surgical MV complexity score previously developed by Anyanwu et al. Methods: Two hundred TEE examinations of patients who underwent MV repair (2009 – 2011) were retrospectively reviewed and scored by two cardiac anesthesiologists. TEE scores were compared to surgical complexity scores of same subset of patients. Kappa values were reported for the agreement of TEE and surgical scores. McNemar’s tests were used to test the homogeneity of the marginal probabilities of different scoring categories. Results: TEE scores were slightly lower (2[1,3]) than surgical scores (3[1,4]). Agreement was 66% between the scoring methods, with a moderate kappa (0.46). Using surgical scores as the gold standard, 70%, 71% and 46% of simple, intermediate and complex surgical scores, respectively, were correctly scored by TEE. P1, P2, P3, and A2 prolapse were easiest to identify with TEE and had the highest agreement with surgical scoring (P1 agreement 79% with kappa 0.55, P2 96% (kappa 0.8), P3 77% (kappa 0.51), A2 88% (kappa 0.6)). The lowest agreement between the two scores occurred with A1 prolapse (kappa 0.05) and posteromedial commissure prolapse (kappa 0.14) (Figure 3). In the presence of significant disagreement, TEE scores were more likely to be of higher complexity than surgical. McNemar’s test was significant for prolapse of P1 (p=0.005), A1 (p=0.025), A2 (p=0.041), and the posteromedial commissure (p<.0001).