Cyclin dependent kinase A; 1 (CDKA; 1) is essential in G1/S transition of cell cycle and its oxidation has been implicated in cell cycle arrest during plant abiotic stress. In the present study, an ...evaluation at the molecular level was performed to find possible sites of protein oxidative modifications. In vivo studies demonstrated that carbonylation of maize CDKA,1 is associated with a decrease in complex formation with maize cyclin D (CycD). Control and in vitro oxidized recombinant CDKA; 1 were sequenced by mass spectrometry. Proline at the PSTAIRE cyclin-binding motif was identified as the most susceptible oxidation site by comparative analysis of the resulted peptides. The specific interaction between CDKA; 1 and CycD6; 1, measured by surface plasmon resonance (SPR), demonstrated that the affinity and the kinetic of the interaction depended on the reduced-oxidized state of the CDKA; 1. CDKA; 1 protein oxidative modification would be in part responsible for affecting cell cycle progression, and thus producing plant growth inhibition under oxidative stress.
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•Carbonylation of cyclin dependent kinase in vivo is associated with a decrease in complex formation with cyclin D (CycD).•CDKA; 1-CycD6; 1 interaction depends on the reduced -oxidized state of the CDKA; 1.•Proline from cyclin –binding motif of CDKA; 1 as a susceptible site for oxidation.
Evaluation of an electrochemical biosensor based on glassy carbon (GC) electrode that was modified with multiwall carbon nanotubes (MWCNTs), which in turn were modified with hydroxyapatite (HAp) and ...horseradish peroxidase (HRP), for the detection of hydrogen peroxide (H2O2), is reported. Hydroxyapatite nanoparticles namely HAp5 and HAp20, that respectively correspond to five- and twenty-days aging times, were in situ synthesised on functionalized carbon nanotubes (fCNTs) walls by a biomimetic procedure using a solution mimicking the inorganic composition of human blood plasma (simulated body fluid (SBF)). HRP was introduced dropwise to the electrode surface. The materials were characterized by transmission electron microscopy, Fourier-transform infrared spectroscopy, X-ray diffraction, thermogravimetry and electrochemical techniques. The Michaelis-Menten apparent constant obtained for both sensors were 0.37 mmol L−1 for HAp5-fCNT/GC and 2.93 mmol L−1 for HAp20-fCNT/GC, indicating that the HRP keeps its enzymatic activity on both electrodes H2O2 detection was performed by chronoamperometry. The linear range obtained for the HRP/HAp5-fCNT/GC electrode and the HRP/HAp20-fCNT/GC electrode was from 1.0 × 10−5 M to 2.34 × 10−4 M, with a detection limit of 1.91 μmol L−1 for the HRP/Hap5-fCNT/GC electrode and 4.45 μmol L−1 for the HRP/HAp20-fCNT/GC electrode. H2O2 was quantified using real-life raw and pasteurized milk samples using the electrode that showed the better performance parameters, the HRP/HAp5-fCNT/GC electrode. The obtained recovery percentage (R%) indicate that the proposed method has a good accuracy.
•An electrochemical biosensor based on carbon nanotubes, hydroxyapatite (Hap) and horseradish peroxidase (HRP) for the detection of hydrogen peroxide is the first time reported.•Electrode modified with Hap nanoparticles with five-days aging times is superior.•Covering of the nanotubes by HAp nanoparticles is one of the main factors that affect the immobilization and the sensitivity of the electrochemical biosensor.
The tetrahydroisoquinoline (THIQ) ring system is present in a large variety of structurally diverse natural products exhibiting a wide range of biological activities. Routes to mimic the biosynthetic ...pathways to such alkaloids, by building cascade reactions in vitro, represents a successful strategy and can offer better stereoselectivities than traditional synthetic methods. S‐Adenosylmethionine (SAM)‐dependent methyltransferases are crucial in the biosynthesis and diversification of THIQs; however, their application is often limited in vitro by the high cost of SAM and low substrate scope. In this study, we describe the use of methyltransferases in vitro in multi‐enzyme cascades, including for the generation of SAM in situ. Up to seven enzymes were used for the regioselective diversification of natural and non‐natural THIQs on an enzymatic preparative scale. Regioselectivites of the methyltransferases were dependent on the group at C‐1 and presence of fluorine in the THIQs. An interesting dual activity was also discovered for the catechol methyltransferases used, which were found to be able to regioselectively methylate two different catechols in a single molecule.
In vitro multi‐enzyme cascade reactions were developed leading to the scalable synthesis of methylated tetra‐hydroisoquinoline alkaloids. The reactions occurred in one‐pot starting from simple building blocks such as dopamine or L‐tyrosine. The use of selected methyltransferases enabled regioselective methylations. When more than one catechol moiety was present, methylations could be directed towards one or more hydroxyl group.
The interest in epigenetic drug and probe discovery is growing as reflected in the large amount of structure-epigenetic activity information available. Therefore, the significance of understanding ...the entire or fractions of the epigenetic relevant chemical space is increasing. Major epigenetic targets are histone lysine deacetylases (HDACs), bromodomains (BRDs), and DNA methyltransferases (DNMTs). However, with the exception of DNMTs, characterization of the chemical space of these epi-targets is limited. This work is the first chemoinformatic analysis of the physicochemical properties, structural diversity, and coverage of the chemical space of compounds screened as inhibitors of HDACs and BRDs. The chemical space was compared to DNMTis, approved drugs, commercial screening compounds, and generally recognized as safe (GRAS) molecules. The structural complexity of compounds directed towards epigenetic targets was also addressed. The outcome of this analysis indicated that it is required to increase the structural diversity and molecular complexity of screening libraries tested as modulators of DNMTs, HDACs and BRDs. Results also suggested that it is feasible to develop dual inhibitors targeting HDACS and BRDs. This work has implications in repurposing of food chemicals with potential epigenetic activity and design of poly-epigenetic compounds.
The management of unsolved inherited retinal dystrophies (IRD) cases is challenging since no standard pipelines have been established. This study aimed to define a diagnostic algorithm useful for the ...diagnostic routine and to address unsolved cases. Here, we applied a Next-Generation Sequencing-based workflow, including a first step of panel sequencing (PS) followed by clinical-exome sequencing (CES) and whole-exome sequencing (WES), in 46 IRD patients belonging to 42 families. Twenty-six likely causal variants in retinal genes were found by PS and CES. CES and WES allowed proposing two novel candidate
(
and a X-linked region including
), both abundantly expressed in human retina according to RT-PCR and immunohistochemistry. After comparison studies, PS showed the best quality and cost values, CES and WES involved similar analytical efforts and WES presented the highest diagnostic yield. These results reinforce the relevance of panels as a first step in the diagnostic routine and suggest WES as the next strategy for unsolved cases, reserving CES for the simultaneous study of multiple conditions. Standardizing this algorithm would enhance the efficiency and equity of clinical genetics practice. Furthermore, the identified candidate genes could contribute to increase the diagnostic yield and expand the mutational spectrum in these disorders.
The recovery and survival of the Macaronesian laurel forest depends on its regeneration strategies. After years of long-term monitoring, both sexual and asexual regeneration appear to be equally ...important. However, the mechanisms for each are just beginning to be understood. In order to contribute to the understanding of the laurel forest sexual regeneration, we analyzed the species composition of the seedling bank every two weeks over three years in the laurel forest of Anaga (Tenerife, Canary Islands). We compared the species compositions of the seedling bank with the canopy, and analyzed changes in their diversity over this period in different forest stands. We found that species diversity (evenness) is different among plots regardless of the stand. In some cases, plot diversity remained constant over time, while others showed some variations, which were little related to climatic conditions (temperature and precipitation). We also found no relationship between the seedling bank and canopy composition, with shade-intolerant species being more abundant in the former. Although climatic conditions remained constant during the period and other environmental conditions did not vary either, some changes were found in the seedling bank species composition. These were related to the increased degree of conservation of the laurel forest of Anaga (by closing unpaved roads, limiting access, and the abandonment of agriculture) that had negatively affected the density of shade-intolerant species. We suggest that such conservation measures should be maintained and extended to other areas where agriculture has been recently abandoned to allow the potential establishment of laurel forest and late successional species.
A global outbreak of the human monkeypox virus (HMPXV), first identified in May 2022, was declared a health emergency of international concern on 23 July 2022. Before the global outbreak, monkeypox ...cases were mostly confined to central and west African countries, where this virus is prevalent. Close contact, mainly sexual contact, is supposed to be the main route of transmission, and it is remarkable that the incidence is higher in men who have sexual relationships with other men.
A 40-year-old Caucasian man arrived at the emergency department complaining of oppressive epigastric pain extending to the chest after a diagnosis of pharyngitis, which was suspected to be caused by the human monkeypox virus. Based on the clinical symptoms, physical examination, serum cardiac biomarkers, and electrocardiographic findings, he was diagnosed with myopericarditis. The real-time PCR for human monkeypox in skin lesions, urine, plasma, and the oropharyngeal swab was positive. The peak of troponin I was 20.6 ng/ml, and the electrocardiogram showed an upward concavity in the ST segment in diffuse leads, which was in agreement with the previous diagnosis. The presence of edema, subepicardial, and myocardial late gadolinium enhancement, and increased values on T1 mapping in the cardiac MRI were in agreement with the diagnosis of myopericarditis. Antiviral treatment with tecovirimat was started with excellent tolerability. After 6 days, the patient recovered and was discharged.
To our knowledge, this is one of the first reported cases of myopericarditis due to human monkeypox infection, which was confirmed by a cardiac MRI following modified Lake Louise criteria. The short span between the onset of the mucocutaneous symptoms and the myocardial damage suggests a pathogenic association. Furthermore, the active viral replication in plasma samples and the negative results on real-time PCR for other viruses support this clinical association.
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•A LIBS methodology for liquid sample analysis was developed and tested.•Common paper sheets are a convenient Ca source for molecular recombination.•Nanosecond-laser ablation is ...capable of breaking strong fluorine-carbon bonds.•Nebulization-assisted LIBS can be implemented for online screening of halogens.
Halogen determination in liquid matrices with Laser-Induced Breakdown Spectroscopy (LIBS) is still a largely unexplored area of research. Together with the intrinsic problematics of liquid samples in LIBS analysis, halogen atomic emission is hard to observe without experimental modifications (e.g. vacuum chambers for VUV resonant lines or controlled He atmospheres for enhanced IR lines) or without switching to indirect determination via halide molecules emission. For the latter, the presence of an alkali-earth metal element is necessary. Previous works have explored CaCO3 pellets as a substrate provider of Ca, and a nebulization-assisted methodology, with great potential for online implementation, has been successfully tested with synthetic and real F-containing samples. However, manufacturing pellets is a time- and reagent-consuming process not suitable for an online analysis technique and a more convenient substrate is needed. In the present work, different kinds of paper, from regular office paper to oil-painting sheets, are tested to explore its feasibility as a Ca-containing solid target. Different sources of F were also considered including inorganic fluoride and fluorocarbon containing samples. Moreover, both fluorine and chlorine determination via molecular emission are considered, achieving limits of detection (LODs) of 5 ppm and 192 ppm, respectively; a common metal (Zn) is also determined via atomic emission to test its validity for multi-elemental microanalysis, with a LOD of 5 ppm.
Lifeguard teams carry out their work in extremely hot conditions in many parts of the world. The aim of this study was to analyze the impact of high temperatures on physiological parameters during ...cardiopulmonary resuscitation (CPR).
A randomized quasi-experimental cross-over design was used to test physiological lifesaving demands (50 min acclimatization +10 min CPR) in two different thermal environments: Thermo-neutral environment (25 °C) vs Hyperthermic environment (37 °C).
The data obtained from 21 lifeguards were included, this covers a total of 420 min of resuscitation. The CPR performance was constantly maintained during the 10 min. The Oxygen uptake (VO 2) ranged from 17 to 18 ml/min/kg for chest compressions (CC) and between 13 and 14 ml/min/kg for ventilations (V) at both 25 °C and 37 °C, with no significant difference between environments (p > 0.05). The percentage of maximum heart rate (%HR max) increased between 7% and 8% at 37 °C (p < 0.001), ranging between 75% and 82% of HR max. The loss of body fluids (LBF) was higher in the hyperthermic environment; LBF: (37 °C: 400 ± 187 g vs 25 °C: 148 ± 81 g, p < 0.001). Body temperature was 1 °C higher at the end of the test (p < 0.001). The perceived fatigue (RPE) increased by 37° an average of 2 points on a scale of 10 (p = 0.001).
Extreme heat is not a limiting factor in CPR performance with two lifeguards. Metabolic consumption is sustained, with an increase in CC, so V can serve as active rest. Nevertheless, resuscitation at 37 °C results in a higher HR, is more exhausting and causes significant loss of fluids due to sweating.
Non-cystic fibrosis bronchiectasis is a chronic structural lung condition that courses with recurrent infectious exacerbations that lead to frequent antibiotic treatment making this population more ...susceptible to acquire pathogens with antibiotic resistance. We aimed to investigate risk factors associated with isolation of multidrug-resistant pathogens in bronchiectasis exacerbations.
A prospective observational study was conducted in two tertiary-care hospitals, enrolling patients when first exacerbation appeared. Multidrug-resistance was determined according to European Centre of Diseases Prevention and Control classification.
Two hundred thirty three exacerbations were included and microorganisms were isolated in 159 episodes. Multidrug-resistant pathogens were found in 20.1% episodes: Pseudomonas aeruginosa (48.5%), methicillin-resistant Staphylococcus aureus (18.2%) and Extended spectrum betalactamase + Enterobacteriaceae (6.1%), and they were more frequent in exacerbations requiring hospitalization (24.5% vs. 10.2%, p: 0.016). Three independent multidrug-resistant risk factors were found: chronic renal disease (Odds ratio (OR), 7.60, 95% CI 1.92-30.09), hospitalization in the previous year (OR, 3.88 95% CI 1.37-11.02) and prior multidrug-resistant isolation (OR, 5.58, 95% CI 2.02-15.46). The proportion of multidrug-resistant in the 233 exacerbations was as follows: 3.9% in patients without risk factors, 12.6% in those with 1 factor and 53.6% if ≥2 risk factors.
Hospitalization in the previous year, chronic renal disease, and prior multidrug-resistant isolation are risk factors for identification multidrug-resistant pathogens in exacerbations. This information may assist clinicians in choosing empirical antibiotics in daily clinical practice.
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