Enzyme Therapy: Current Challenges and Future Perspectives

In recent years, enzymes have risen as promising therapeutic tools for different pathologies, from metabolic deficiencies, such as fibrosis conditions, ocular pathologies or joint problems, to cancer or cardiovascular diseases. Treatments based on the catalytic activity of enzymes are able to convert a wide range of target molecules to restore the correct physiological metabolism. These treatments present several advantages compared to established therapeutic approaches thanks to their affinity and specificity properties. However, enzymes present some challenges, such as short in vivo half-life, lack of targeted action and, in particular, patient immune system reaction against the enzyme. For this reason, it is important to monitor serum immune response during treatment. This can be achieved by conventional techniques (ELISA) but also by new promising tools such as microarrays. These assays have gained popularity due to their high-throughput analysis capacity, their simplicity, and their potential to monitor the immune response of patients during enzyme therapies. In this growing field, research is still ongoing to solve current health problems such as COVID-19. Currently, promising therapeutic alternatives using the angiotensin-converting enzyme 2 (ACE2) are being studied to treat COVID-19.This research was funded by the Basque Government, BIKAINTEK, grant number 48-AF-W2-2019-00006; and by University of the Basque Country, PIFIND19/02, grant number 201900016247

Lisofosfolipidoen eta Alzheimer gaixotasunaren arteko erlazioa: etorkizuneko itu farmakologikoaren bila

Lipidoak funtzio energetikoaz eta egitura-funtzioez gain deskribatu diren beste funtzioei esker garrantzitsuak bilakatzen ari dira. Funtzio neurotransmisorea edota neuromoduladorea aurkeztu duten lipidoen artean, lisofosfolipidoak aurkitu ditzakegu. Lisofosfolipidoak lipido molekula txiki bioaktiboak, karbonodun kate bakarra eta buru polar bat edukitzeagatik bereizten direnak dira. Lisofosfolipidoen artean, azido lisofosfatidikoa eta esfingosina 1-fosfatoaren egitura eta sistemen funtzioak izan dira hobeto deskribatu direnak. Lisofosfolipidoak zelulaz kanpoko bitartekari aritzen dira berentzat espezifikoak diren G proteinei loturiko hartzaileak aktibatuz. Molekula horien seinaleztapenaren bidez zenbait prozesu neurokimiko modulatzen dira, adibidez, neuromodulazioa eta neuroinflamazioa. Gainera, ikasketarekin eta oroimenarekin erlazioa erakutsi dute. Horren haritik, orain arte ondoen deskribatutako lisofosfolipidoen sistemak, azido lisofosfatidikoa eta esfingosina l-fosfatoa, hain zuzen ere, asaldatuta daude Alzheimer gaixotasunean eta gaixotasun honetako zenbait animalia-eredutan. Aldaketa horien zentzua oraindik ez dago finkatuta, baina haien eragina beste neurotransmisio-sistemen edota bestelako funtzio biologikoen modulazioaren bidez gerta daitezke. Beraz, lipido hauek etorkizun handiko itu farmakologikoak izan daitezke Alzheimer gaixotasunean agertzen diren sintoma neuropatologikoak eta neuropsikiatrikoak arintzeko. Hortaz, merkatuan dauden lipidoen seinaleztapena itutzat duten eta beste neuroendekapenezko gaixotasunak tratatzeko erabilgarriak diren farmakoak baliagarriak izan litezke Alzheimer gaixotasuna tratatzeko, aukera emanez horrela Alzheimer gaixotasuna tratatzeko dagoen hutsune farmakologikoa betetzeko.; In addition to energy and structural functions, lipids are becoming important thanks to the other functions described. Some lipids have been shown to exhibit neurotransmitter or neuromodulatory function, including lysophospholipids. Lysophospholipids are small bioactive lipid molecules that are distinguished only by having a single carbon chain and a single polar head. The lysophosphatidic acid and sphingosine l-phosphate structure and system functions are best described among those with neurotransmitter function. Lysophospholipids act as extracellular mediators that activate receptor-specific G proteins that are specific to them. The signaling of these molecules modulate certain neurochemical processes, including neuromodulation and neuroinflammation. They have also presented the relationship with learning and memory. In this respect, the best described lysophospholipid systems, lysophosphatidic acid and sphingosine 1-phosphate, are indeed disturbed in Alzheimer’s disease and in some animal models of this disease. The meaning of these changes is not yet established, but their effect may be related to the modulation of other neurotransmission systems or other biological functions. These lipids are therefore supposed to be the promising pharmacological targets to alleviate the neuropathological and neuropsychiatric symptoms that appear in Alzheimer’s disease. Therefore, marketed drugs that have lipid signaling as a pharmacological target and that are useful to treat other neurodegenerative diseases could be also helpful to treat the Alzheimer’s disease, and with this it might be possible to fill the pharmacological gap in the treatment of Alzheimer’s disease so far

Creación del primer Grupo iGEM (Competición Internacional de Biología Sintética) de Madrid: Facultad de Biología-UCM

En este Trabajo se describe la creación del primer Grupo iGEM de Madrid así como la elaboración de un biosensor que permite la detección de polen de olivo a tiempo real empleando metodos de ingeniería genética y robótica

Allergenic Lipid Transfer Proteins from Plant-Derived Foods Do Not Immunologically and Clinically Behave Homogeneously: The Kiwifruit LTP as a Model

BACKGROUND: Food allergy is increasingly common worldwide. Tools for allergy diagnosis measuring IgE improved much since allergenic molecules and microarrays started to be used. IgE response toward allergens belonging to the same group of molecules has not been comprehensively explored using such approach yet. OBJECTIVE: Using the model of lipid transfer proteins (LTPs) from plants as allergens, including two new structures, we sought to define how heterogeneous is the behavior of homologous proteins. METHODS: Two new allergenic LTPs, Act d 10 and Act c 10, have been identified in green (Actinidia deliciosa) and gold (Actinidia chinensis) kiwifruit (KF), respectively, using clinically characterized allergic patients, and their biochemical features comparatively evaluated by means of amino acid sequence alignments. Along with other five LTPs from peach, mulberry, hazelnut, peanut, mugwort, KF LTPs, preliminary tested positive for IgE, have been immobilized on a microarray, used for IgE testing 1,003 allergic subjects. Comparative analysis has been carried out. RESULTS: Alignment of Act d 10 primary structure with the other allergenic LTPs shows amino acid identities to be in a narrow range between 40 and 55%, with a number of substitutions making the sequences quite different from each other. Although peach LTP dominates the IgE immune response in terms of prevalence, epitope recognition driven by sequence heterogeneity has been recorded to be distributed in a wide range of behaviors. KF LTPs IgE positive results were obtained in a patient subset IgE positive for the peach LTP. Anyhow, the negative results on homologous molecules allowed us to reintroduce KF in patients' diet. CONCLUSION: The biochemical nature of allergenic molecule belonging to a group of homologous ones should not be taken as proof of immunological recognition as well. The availability of panels of homologous molecules to be tested using microarrays is valuable to address the therapeutic intervention

Robust estimation of bacterial cell count from optical density

Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals <1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data

Endocannabinoid and Muscarinic Signaling Crosstalk in the 3xTg-AD Mouse Model of Alzheimer's Disease

The endocannabinoid system, which modulates emotional learning and memory through CB 1 receptors, has been found to be deregulated in Alzheimer's disease (AD). AD is characterized by a progressive decline in memory associated with selective impairment of cholinergic neurotransmission. The functional interplay of endocannabinoid and muscarinic signaling was analyzed in seven-month-old 3xTg-AD mice following the evaluation of learning and memory of an aversive stimulus. Neurochemical correlates were simultaneously studied with both receptor and functional autoradiography for CB 1 and muscarinic receptors, and regulations at the cellular level were depicted by immunofluorescence. 3xTg-AD mice exhibited increased acquisition latencies and impaired memory retention compared to age-matched non-transgenic mice. Neurochemical analyses showed changes in CB 1 receptor density and functional coupling of CB 1 and muscarinic receptors to G i/o proteins in several brain areas, highlighting that observed in the basolateral amygdala. The subchronic (seven days) stimulation of the endocannabinoid system following repeated WIN55,212-2 (1 mg/kg) or JZL184 (8 mg/kg) administration induced a CB 1 receptor downregulation and CB 1 -mediated signaling desensitization, normalizing acquisition latencies to control levels. However, the observed modulation of cholinergic neurotransmission in limbic areas did not modify learning and memory outcomes. A CB 1 receptor-mediated decrease of GABAergic tone in the basolateral amygdala may be controlling the limbic component of learning and memory in 3xTg-AD mice. CB 1 receptor desensitization may be a plausible strategy to improve behavior alterations associated with genetic risk factors for developing AD.SCOPUS: ar.jinfo:eu-repo/semantics/publishe

The biomechanics of wounds at physiologically relevant levels: understanding skin’s stress-shielding effect for the quantitative assessment of healing

Wounds are responsible for the decrease in quality of life of billions of people around the world. Their assessment relies on subjective parameters which often delays optimal treatments and results in increased healthcare costs. In this work, we sought to understand and quantify how wounds at different healing stages (days 1, 3, 7 and 14 post wounding) change the mechanical properties of the tissues that contain them, and how these could be measured at clinically relevant strain levels, as a step towards quantitative wound tracking technologies. To achieve this, we used digital image correlation and mechanical testing on a mouse model of wound healing to map the global and local tissue strains. We found no significant differences in the elastic and viscoelastic properties of wounded vs unwounded skin when samples were measured in bulk, presumably as these were masked by the protective mechanisms of skin, which redistributes the applied loads to mitigate high stresses and reduce tissue damage. By measuring local strain values and observing the distinct patterns they formed, it was possible to establish a connection between the healing phase of the tissue (determined by the time post-injury and the observed histological features) and the overall mechanical behaviour. Importantly, these parameters were measured from the surface of the tissue, using physiologically relevant strains without increasing the tissue's damage. Adaptations of these approaches for clinical use have the potential to aid in the identification of skin healing problems, such as excessive inflammation or lack of mechanical progression over time. An increase, decrease, or lack of change in the elasticity and viscoelasticity parameters, can be indicative of wound state, thus ultimately leading to improved diagnostic outcomes

Specific Phospholipid Modulation by Muscarinic Signaling in a Rat Lesion Model of Alzheimers Disease

[Image: see text] Alzheimer’s disease (AD) represents the most common cause of dementia worldwide and has been consistently associated with the loss of basal forebrain cholinergic neurons (BFCNs) leading to impaired cholinergic neurotransmission, aberrant synaptic function, and altered structural lipid metabolism. In this sense, membrane phospholipids (PLs) can be used for de novo synthesis of choline (Ch) for the further obtaining of acetylcholine (ACh) when its availability is compromised. Specific lipid species involved in the metabolism of Ch have been identified as possible biomarkers of phenoconversion to AD. Using a rat model of BFCN lesion, we have evaluated the lipid composition and muscarinic signaling in brain areas related to cognitive processes. The loss of BFCN resulted in alterations of varied lipid species related to Ch metabolism at nucleus basalis magnocellularis (NMB) and cortical projection areas. The activity of muscarinic receptors (mAChR) was decreased in the NMB and increased in the hippocampus according to the subcellular distribution of M(1)/M(2) mAChR which could explain the learning and memory impairment reported in this AD rat model. These results suggest that the modulation of specific lipid metabolic routes could represent an alternative therapeutic strategy to potentiate cholinergic neurotransmission and preserve cell membrane integrity in AD

On the Use of Graphene to Improve the Performance of Concentrator III-V Multijunction Solar Cells

Graphene has been intensively studied in photovoltaics focusing on emerging solar cells based on thin films, dye-sensitized, quantum dots, nanowires, etc. However, the typical efficiency of these solar cells incorporating graphene are below 16%. Therefore, the photovoltaic potential of graphene has not been already shown. In this work the use of graphene for concentration applications on III-V multijunction solar cells, which indeed are the solar cells with the highest efficiency, is demonstrated. Firstly, a wide optoelectronic characterization of graphene layers is carried out. Then, the graphene layer is incorporated onto triple-junction solar cells, which decreases their series resistance by 35% (relative), leading to an increase in Fill Factor of 4% (absolute) at concentrations of 1,000 suns. Simultaneously, the optical absorption of graphene produces a relative short circuit current density decrease in the range of 0-1.8%. As a result, an absolute efficiency improvement close to 1% at concentrations of 1,000 suns was achieved with respect to triple junction solar cells without graphene. The impact of incorporating one and two graphene monolayers is also evaluated