Publications by authors named "Alcalde M"

Galactose oxidase (GOase) is a versatile biocatalyst with a wide range of potential applications, ranging from synthetic chemistry to bioelectrochemical devices. Previous GOase engineering by directed evolution generated the M-RQW mutant, with unprecedented new-to-nature oxidation activity at the C6-OH group of glucose, and a mutational backbone that helped to unlock its promiscuity toward other molecules, including secondary alcohols. In the current study, we have used the M-RQW mutant as a starting point to engineer a set of GOases that are very thermostable and that are easily produced at high titers in yeast, enzymes with latent activities applicable to sustainable chemistry.

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The influence of ultrasonic activation (UA) on the physicochemical properties of setting time (n = 5), volumetric stability (n = 10), alkalizing activity/calcium ions release (pH/Ca) (n = 10) and calcium/phosphate-rich substance formation in simulated body fluid (Hank's balanced salt solution; HBSS) of three cements was evaluated: white MTA-Angelus (MTA), MTA Repair HP (MTAHP) and Biodentine (BIO). It was observed that UA influenced the setting times (min) distinctly, delaying the initial setting time of MTA (40.8/64.

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Plastic waste is a major threat in our industrialized world and is driving research into bioplastics. The success of biobased polyethylene furanoate (PEF) as a viable alternative to polyethylene terephthalate (PET) of fossil origin will depend on designing effective enzymes to break it down, aiding its recycling. Here, a panel of fungal and bacterial cutinases were functionally expressed in a tandem yeast expression system based on and .

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Enzymes that oxidize aromatic substrates have shown utility in a range of cell-based technologies including live cell proximity labeling (PL) and electron microscopy (EM), but are associated with drawbacks such as the need for toxic HO. Here, we explore laccases as a novel enzyme class for PL and EM in mammalian cells. LaccID, generated via 11 rounds of directed evolution from an ancestral fungal laccase, catalyzes the one-electron oxidation of diverse aromatic substrates using O instead of toxic HO, and exhibits activity selective to the surface plasma membrane of both living and fixed cells.

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Fungal unspecific peroxygenases (UPOs) are gaining momentum in synthetic chemistry. Of special interest is the UPO from (UPO), which shows an exclusive repertoire of oxyfunctionalizations, including the terminal hydroxylation of alkanes, the α-oxidation of fatty acids and the C-C cleavage of corticosteroids. However, the lack of heterologous expression systems to perform directed evolution has impeded its engineering for practical applications.

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Arrhythmogenic cardiomyopathy is an inherited entity characterized by irregular cell-cell adhesion, cardiomyocyte death and fibro-fatty replacement of ventricular myocytes, leading to malignant ventricular arrythmias, contractile dysfunction and sudden cardiac death. Pathogenic variants in genes that encode desmosome are the predominant cause of arrhythmogenic cardiomyopathy. Moreover, signalling pathways such as Wnt/ß-catenin and transforming growth factor-β have been involved in the disease progression.

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The siderophore-cephalosporin cefiderocol (FDC) presents a promising treatment option for carbapenem-resistant (CR) (PA). FDC circumvents traditional porin and efflux-mediated resistance by utilizing TonB-dependent receptors (TBDRs) to access the periplasmic space. Emerging FDC resistance has been associated with loss of function mutations within TBDR genes or the regulatory genes controlling TBDR expression.

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Background: Spastic paraplegia 11 (SPG11) is the most prevalent form of autosomal recessive hereditary spastic paraplegia, resulting from biallelic pathogenic variants in the SPG11 gene (MIM *610844).

Methods: The proband is a 36-year-old female referred for genetic evaluation due to cognitive dysfunction, gait impairment, and corpus callosum atrophy (brain MRI was normal at 25-years-old). Diagnostic approaches included CGH array, next-generation sequencing, and whole transcriptome sequencing.

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This study aimed to investigate obesity-related glomerulopathy (ORG) at cellular, structural, and transcriptomic levels. Thirty Wistar rats were randomized into two groups: 15 rats were fed with a standard diet (SD-rats), and 15 rats were fed with a high-fat diet (HFD-rats). After 10 weeks, the weight, kidney function, histological features, and transcriptomic changes were assessed.

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Introduction: The purpose of this study was to evaluate whether the mixing method of AH Plus Jet sealer affects its physicochemical and mechanical properties.

Methods: The properties of AH Plus Jet sealer were analyzed when mixed using either the Auto Mix Tip or manual mixing. The evaluated properties included radiopacity (n = 5), initial and final setting times (n = 5), flow (n = 5), and solubility (n = 3), following specifications outlined in ISO 6876/2012 and ADA Nº 57/2000.

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Objectives: This study evaluated the impact of different methods of irrigant agitation on smear layer removal in the apical third of curved mesial canals of 3 dimensionally (D) printed mandibular molars.

Materials And Methods: Sixty 3D-printed mandibular second molars were used, presenting a 70° curvature and a Vertucci type II configuration in the mesial root. A round cavity was cut 2 mm from the apex using a trephine of 2 mm in diameter, 60 bovine dentin disks were made, and a smear layer was formed.

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Background: Symptomatic irreversible pulpitis often results in heightened reactions to thermal stimuli such as pain evoked by a cold stimulus, and spontaneous odontogenic pain (unprovoked pain).

Objective: This study primarily compared the clinical manifestations of odontogenic spontaneous pain and pain provoked by cold stimulus specifically focusing on their sensory discriminative characteristics (intensity, duration and quality) between symptomatic irreversible pulpitis patients with and without referred pain.

Methods: Twenty-three patients with symptomatic irreversible pulpitis with referred pain and 12 patients without referred pain were included in this cross-sectional study.

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Article Synopsis
  • The study investigates how the strength of temporary dental restorations is affected by thermal aging, focusing on different resin-based materials.
  • It involves testing 192 resin specimens made from various materials under thermal cycling conditions to assess their flexibility and strength.
  • Results show that the type of resin material and the thermal aging process significantly influence their strength, with Filtek Bulk-Fill performing better than J-Temp and Fuji Lining.
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Objective:  The aim of this study was to evaluate the accuracy of the Root ZX II (RZX), Raypex 6 (RAY), EPex Pro (EPEX), and CanalPro (CNP) electronic foramen locators (EFLs) in different foraminal morphologies (fully formed foramen, immature foramen with parallel walls, and immature foramen with divergent walls); this article also evaluated the influence of different penetration levels (0.0 mm and -1.0 mm).

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Article Synopsis
  • The study analyzed 10 years of data on adult patients with solid tumors enrolled in clinical trials, focusing on the clinical characteristics and the financial impact of sponsor contributions to drug costs.
  • Over the decade, the number of clinical trials increased significantly from 140 in 2010 to 459 in 2019, with a shift in drug use from mainly targeted therapies to a greater emphasis on immunotherapy.
  • The analysis showed that while the overall expenditure on antineoplastic drugs outside clinical trials was over EUR 120 million, sponsor contributions in trial settings amounted to about EUR 107 million, resulting in a potential cost avoidance of around EUR 92 million.
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Article Synopsis
  • Dilated cardiomyopathy is a complex heart condition linked to heart failure and arrhythmias, with about 50% of cases being inherited, highlighting the importance of genetic analysis.
  • In a study of 65 patients, researchers re-evaluated 125 previously ambiguous genetic variants, reclassifying 12% as likely benign or pathogenic, suggesting improved understanding through better population frequency data.
  • Their new algorithm indicated that 60.9% of remaining ambiguous variants had potential but unconfirmed harmful effects, while 24.5% might be benign, demonstrating the benefit of regularly updating genetic data for clearer interpretations.
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Objectives: To perform a systematic review of animal studies that compared the histopathological characteristics between teeth with apical periodontitis after endodontic treatment in one or two visits.

Materials And Methods: This systematic review was registered on the International Prospective Register of Systematic Reviews (PROSPERO) - CRD42022340849. Studies were collected from PubMed/MEDLINE, LILACS, EMBASE, Livivo, SciELO, Web of Science, Scopus, and Cochrane Library and manual and gray literature searches.

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Introduction: This study assessed the effect of intentional foraminal enlargement on the foramen and the apical root canal morphology.

Method: Sixty mesial roots of mandibular molars were scanned by micro-computed tomography. Their apical foramina were photographed with a stereomicroscope before and after preparation.

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The siderophore-cephalosporin cefiderocol(FDC) presents a promising treatment option for carbapenem-resistant (CR) (PA). FDC circumvents traditional porin and efflux mediated resistance by utilizing TonB-dependent receptors (TBDRs) to access the periplasmic space. Emerging FDC resistance has been associated with loss of function mutations within TBDR genes or the regulatory genes controlling TBDR expression.

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Biomaterials are used extensively in graft procedures to correct bone defects, interacting with the body without causing adverse reactions. The aim of this pre-clinical study was to analyze the effects of photobiomodulation therapy (PBM) with the use of a low-level laser in the repair process of bone defects filled with inorganic matrix (IM) associated with heterologous fibrin biopolymer (FB). A circular osteotomy of 4 mm in the left tibia was performed in 30 Wistar male adult rats who were randomly divided into three groups: G1 = IM + PBM, G2 = IM + FB and G3 = IM + FB + PBM.

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Introduction: The aim of this study was to assess the effectiveness of filling removal material from the apical third of curved mesial root canals of mandibular molars. Reciprocating instrumentation followed by additional rotary instrumentation with instruments made of alloys with different heat treatments was evaluated.

Materials And Methods: Thirty-six mesial roots of mandibular molars were divided into two groups: Group Class IV consisted of 16 roots with two independent canals, and Group Class II consisted of 20 roots with two canals that merged into one at their apical level.

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Unlabelled: Evaluate the shaping ability and preparation time using a pediatric motor-driven rotary instrumentation compared to other systems in resin prototypes of primary molars.

Methods: Thirty specimens were scanned in micro-CT and divided into three groups according to the instrumentation type: pediatric motor-driven Sequence baby File (SBF); conventional motor-driven (Sequence Rotary File - SRF); manual K file. Instrumentation time was timed.

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Article Synopsis
  • The study examined the effectiveness of the iVac® endodontic irrigation system compared to traditional and ultrasonic irrigation methods on cleaning root canals and dissolving organic tissue.
  • Thirty mandibular premolars were analyzed using scanning electron microscopy to measure the cleaning efficacy across different irrigation techniques, including conventional, ultrasonic, and the iVac® system.
  • Results showed that both the iVac® and ultrasonic methods were more effective than conventional irrigation in cleaning root canal walls and dissolving tissue, with no significant difference between the two advanced methods.
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Fungal unspecific peroxygenases (UPOs) are arising as versatile biocatalysts for C-H oxyfunctionalization reactions. In recent years, several directed evolution studies have been conducted to design improved UPO variants. An essential part of this protein engineering strategy is the design of reliable colorimetric high-throughput screening (HTS) assays for mutant library exploration.

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Recent advancements in plant biotechnology have highlighted the potential of hairy roots as a biotechnological platform, primarily due to their rapid growth and ability to produce specialized metabolites. This study aimed to delve deeper into hairy root development in and explore the optimization of genetic transformation for enhanced bioactive compound production. Previously established hairy root lines of were categorized based on their centelloside production capacity into HIGH, MID, or LOW groups.

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