Advances in the screening of antimicrobial compounds using electrochemical biosensors: is there room for nanomaterials?

The abusive use of antimicrobial compounds and the associated appearance of antimicrobial resistant strains are a major threat to human health. An improved antimicrobial administration involves a faster diagnosis and detection of resistances. Antimicrobial susceptibility testing (AST) are the reference techniques for this purpose, relying mainly in the use of culture techniques. The long time required for analysis and the lack of reproducibility of these techniques have fostered the development of high-throughput AST methods, including electrochemical biosensors. In this review, recent electrochemical methods used in AST have been revised, with particular attention on those used for the evaluation of new drug candidates. The role of nanomaterials in these biosensing platforms has also been questioned, inferring that it is of minor importance compared to other applications.

Electrochemical monitoring of enzymatic cleavage in nanochannels with nanoparticle-based enhancement: determination of MMP-9 biomarker.

For the first time the use of nanoparticles as carriers of an enzymatic substrate immobilized inside nanoporous alumina membranes is proposed with the aim of amplifying the nanochannel blocking produced and, consequently, improving the efficiency of an enzyme determination through enzymatic cleavage. Streptavidin-modified polystyrene nanoparticles (PSNPs) are proposed as carrier agents, contributing to the steric and the electrostatic blockage due to the charge they present at different pH values. Electrostatic blockage is the predominant effect that governs the blockage in the interior of the nanochannel and is dependent not just in the charge inside the channel, but also in the polarity of the redox indicator used. Hence, the effect of using negatively charged ([Fe(CN)6]4-) and positively charged ([Ru(NH3)6]3+) redox indicator ions is studied for the first time. Under the optimum conditions, matrix-metalloproteinase 9 (MMP-9) is detected at clinically relevant levels (100-1200 ng/mL) showing a detection limit of 75 ng/mL and a quantification limit of 251 ng/mL with good reproducibility (RSD: 8%) and selectivity, also showing an excellent performance in real samples with acceptable recovery percentages (in the range around 80-110%). Overall, our approach represents a cheap and fast sensing methodology of great potential in point-of-care diagnostics.

Coping strategies and stress-induced natural killer cell redistribution in women with eating disorders.

BACKGROUND: Patients with eating disorders (ED) are very sensitive and responsive to psychosocial stress. Stress response includes changes in immune cell distribution and may be modulated by the capability to cope with stressors. Thus, the present study sought to analyze the association between coping strategies and immune response (natural killer [NK] cell redistribution following psychosocial stress) in patients with anorexia nervosa (AN) and bulimia nervosa (BN) and healthy controls (HC). METHOD: Twenty-four AN patients, 29 BN patients, and 58 HC were studied. A multidimensional assessment tool, the COPE Inventory, was used to assess coping strategies. The number of NK cells was quantified in peripheral blood before and after the application of the Trier Social Stress Test (TSST). Potentially mediating variables, such as weight status, severity of eating pathology, depression, anxiety, and impulsivity were controlled. RESULTS: The three groups differed in intensity and direction of cell redistribution: The TSST was followed in BN patients by a significant decrease in the number of NK cells, whereas HC displayed a moderate decrease and AN a clear increase. Specific correlations between coping strategies and NK cell mobilization were found, especially in BN patients (positive for "planning" and negative for "substance abuse"). CONCLUSION: Recognition and subsequent modification of the dysfunctional coping strategies used by patients with ED could contribute to improving their immune status, strengthening their resilience and increasing their ability to overcome the disease.

ANTECEDENTES: Los pacientes con trastornos de la conducta alimentaria (TCA) son muy sensibles y respondedores ante el estrés psicosocial. La respuesta al estrés incluye cambios en la distribución de las células inmunes y parece estar modulada por la capacidad de afrontamiento del individuo. En este contexto, el objetivo del presente estudio fue analizar la asociación entre estrategias de afrontamiento y redistribución tras el estrés de las células agresoras naturales (natural killer, NK) en pacientes de sexo femenino con anorexia nervosa (AN) y bulimia nervosa (BN), y controles sanos (CS). MÉTODO: Treinta y cuatro pacientes con AN, 29 pacientes con BN y 58 CS fueron estudiados. Para evaluar las estrategias de afrontamiento se utilizó un instrumento multidimensional, el Inventario COPE. El número de células NK en sangre periférica fue cuantificado antes y después de la aplicación del Trier Social Stress Test (TSST). Las posibles variables mediadoras, como el estado ponderal, la gravedad de la patología alimentaria, depresión, ansiedad e impulsividad fueron control. RESULTADOS: Los tres grupos difirieron en la intensidad y la dirección de la redistribución de células NK: el TSST fue seguido de una notable reducción en el número de células NK en las pacientes con BN, de una disminución moderada en las CS y de un claro incremento en las pacientes con AN. Se encontraron correlaciones específicas entre estrategias de afrontamiento y movilización de células NK, especialmente en las pacientes con BN (positiva para "planificación" y negativa para "abuso de sustancias"). CONCLUSIÓN: El reconocimiento y la modificación consiguiente de las estrategias de afrontamiento disfuncionales utilizadas por las pacientes con TCA pueden contribuir a mejorar su estado inmunológico, incrementando su resistencia y su capacidad para superar la enfermedad.

Electrochemical Biosensors Based on Nanomaterials for Early Detection of Alzheimer's Disease.

Alzheimer's disease (AD) is an untreatable neurodegenerative disease that initially manifests as difficulty to remember recent events and gradually progresses to cognitive impairment. The incidence of AD is growing yearly as life expectancy increases, thus early detection is essential to ensure a better quality of life for diagnosed patients. To reach that purpose, electrochemical biosensing has emerged as a cost-effective alternative to traditional diagnostic techniques, due to its high sensitivity and selectivity. Of special relevance is the incorporation of nanomaterials in biosensors, as they contribute to enhance electron transfer while promoting the immobilization of biological recognition elements. Moreover, nanomaterials have also been employed as labels, due to their unique electroactive and electrocatalytic properties. The aim of this review is to add value in the advances achieved in the detection of AD biomarkers, the strategies followed for the incorporation of nanomaterials and its effect in biosensors performance.

Long term effects on biochemical bone markers of a single infusion of zoledronic acid in Paget disease of bone.

BACKGROUND: The aim of the present study is to evaluate long term biochemical response to a single dose of zoledronic acid in patients with Paget disease of bone, as well as evaluating the value of bone turnover markers in diagnosis and follow-up. METHODS: This is an observational, descriptive and prospective study. Included patients received a single-dose intravenous infusion of 5 mg zoledronic acid. Bone turnover markers were measured at baseline, and in every follow up visit. RESULTS: Thirty-nine patients with a mean follow-up of 56.49 months were included. At the time Paget disease was diagnosed, all of the patients (100%) had high serum procollagen type 1 amino-terminal propeptide values, but not all patients had high serum C-terminal telopeptide and alkaline phosphatase values (85% and 89% respectively). Biochemical response to therapy occurred in 38 out of 39 patients (97%). Two patients had partial response at 6 months but complete response thereafter. Only one patient relapsed (nadir procollagen type 1 amino-terminal propeptide 35.06 µg/l, value at relapse 75.2 µg/l) 4.5 years after treatment. Values of serum C-terminal telopeptide and alkaline phosphatase of this patient were normal despite P1NP relapse. CONCLUSIONS: We hence conclude that zoledronic acid is effective in inducing and maintaining biochemical remission and that procollagen type 1 amino-terminal propeptide is a better diagnostic and prognostic marker in PDB when compared to C-terminal telopeptide and alkaline phosphatase.

Mutations in Either TUBB or MAPRE2 Cause Circumferential Skin Creases Kunze Type.

Circumferential skin creases Kunze type (CSC-KT) is a specific congenital entity with an unknown genetic cause. The disease phenotype comprises characteristic circumferential skin creases accompanied by intellectual disability, a cleft palate, short stature, and dysmorphic features. Here, we report that mutations in either MAPRE2 or TUBB underlie the genetic origin of this syndrome. MAPRE2 encodes a member of the microtubule end-binding family of proteins that bind to the guanosine triphosphate cap at growing microtubule plus ends, and TUBB encodes a ß-tubulin isotype that is expressed abundantly in the developing brain. Functional analyses of the TUBB mutants show multiple defects in the chaperone-dependent tubulin heterodimer folding and assembly pathway that leads to a compromised yield of native heterodimers. The TUBB mutations also have an impact on microtubule dynamics. For MAPRE2, we show that the mutations result in enhanced MAPRE2 binding to microtubules, implying an increased dwell time at microtubule plus ends. Further, in vivo analysis of MAPRE2 mutations in a zebrafish model of craniofacial development shows that the variants most likely perturb the patterning of branchial arches, either through excessive activity (under a recessive paradigm) or through haploinsufficiency (dominant de novo paradigm). Taken together, our data add CSC-KT to the growing list of tubulinopathies and highlight how multiple inheritance paradigms can affect dosage-sensitive biological systems so as to result in the same clinical defect.

The psychological well-being index and quality of life after a cardiac rehabilitation program based on aerobic training and psychosocial support.

BACKGROUND: This study aimed to investigate the physical and psychological benefits of an alternative cardiac rehabilitation program based on therapeutic groups during physical exercise sessions and to compare the results with those of a conventional cardiac rehabilitation program. METHOD: The sample included 112 patients from the cardiac rehabilitation unit of a medical center, 91.1 % of whom were male. The control group consisted of 47 subjects, with a mean age of 57.89 ± 12.30 and the experimental group consisted of 65 subjects, with a mean age of M = 58.38 ± 9.86. Quality of life, psychological well-being, health-related quality of life, body mass index, blood pressure, abdominal circumference and resting heart rate were measured before starting and at the end of the cardiac rehabilitation program. RESULTS: The experimental group improved significantly more than the control group in body mass index, systolic and diastolic blood pressure, abdominal circumference, and resting heart rate (p value < 0.005). In addition, the experimental group had significantly greater improvements in quality of life, psychological well-being, and health-related quality of life than the control group (p-value < 0.001). CONCLUSIONS: A cardiac rehabilitation program based on simultaneous aerobic training and psychosocial support improved the physical function, health-related quality of life and well-being.

Rapid SARS-CoV-2 sensing through oxygen reduction reaction catalysed by Au@Pt/Au core@shell nanoparticles.

The development of rapid, accurate, sensitive, and low-cost diagnostic methods for COVID-19 detection in real-time is the unique way to control infection sources and monitor illness progression. In this work, we propose an electrochemical biosensor for the rapid and accuracy diagnosis of COVID-19, through the determination of ORF1ab specific sequence. The biosensor is based on the immobilization of a thiolated sequence partially complementary (domain 1) to ORF1ab on gold screen-printed electrodes and the use of bifunctional Au@Pt/Au core@shell nanoparticles modified with a second thiolated sequence partially complementary to ORF1ab (domain 2) as electrochemical indicator of the hybridization of DNA sequences. The synthesized Au@Pt/Au nanoparticles consist of an Au core, a shell of Pt (Au@Pt NPs), that provides an excellent electrocatalytic activity toward the oxygen reduction reaction (ORR) even after formation of hybrid biomaterials by modification, through the Au protuberances growth on the NPs surface, with an oligonucleotide with recognition ability. The ORR electrochemical activity, enhanced by the label element (Au@Pt/Au NPs), has been employed, for the first time, as indicator of the hybridization event. Based on this strategy, target sequences of the SARS-CoV-2 virus have been detected with a detection limit of 32 pM. The selectivity of the biosensor was confirmed by analysing ORF1ab sequence in the presence of DNA sequences from other viruses. The biosensor has been successfully applied to the direct detection of the virus in non-amplified samples of nasopharyngeal swabs from infected and non-infected patients. Results compare well with those obtained through RT-qPCR but our method is more rapid since does not need any amplification process.

Next-generation sequencing reveals that miR-16-5p, miR-19a-3p, miR-451a, and miR-25-3p cargo in plasma extracellular vesicles differentiates sedentary young males from athletes.

A sedentary lifestyle and Olympic participation are contrary risk factors for global mortality and incidence of cancer and cardiovascular disease. Extracellular vesicle miRNAs have been described to respond to exercise. No molecular characterization of young male sedentary people versus athletes is available; so, our aim was to identify the extracellular vesicle miRNA profile of chronically trained young endurance and resistance male athletes compared to their sedentary counterparts. A descriptive case-control design was used with 16 sedentary young men, 16 Olympic male endurance athletes, and 16 Olympic male resistance athletes. Next-generation sequencing and RT-qPCR and external and internal validation were performed in order to analyze extracellular vesicle miRNA profiles. Endurance and resistance athletes had significant lower levels of miR-16-5p, miR-19a-3p, and miR-451a compared to sedentary people. Taking all together, exercise-trained miRNA profile in extracellular vesicles provides a differential signature of athletes irrespective of the type of exercise compared to sedentary people. Besides, miR-25-3p levels were specifically lower in endurance athletes which defines its role as a specific responder in this type of athletes. In silico analysis of this profile suggests a role in adaptive energy metabolism in this context that needs to be experimentally validated. Therefore, this study provides for the first time basal levels of circulating miRNA in extracellular vesicles emerge as relevant players in intertissue communication in response to chronic exercise exposure in young elite male athletes.

Nitrogen-dependent calcineurin activation in the yeast Hansenula polymorpha.

Non-preferred nitrogen sources, unlike preferred ones, raised total cell Ca(2+) content and expression of ENA1, a very well-known calcineurin-regulated gene. This indicates calcineurin activation is regulated by nitrogen source. Nitrogen catabolite repression (NCR) and nitrate induction mechanisms, both regulating nitrate assimilation in Hansenula polymorpha, are controlled by calcineurin. Concerning NCR, lack of calcineurin (cnb1 mutant) decreased nitrate-assimilation gene expression, levels of the transcription factor Gat1 and growth in several nitrogen sources. We found that the role of calcineurin in NCR was mediated by Crz1 via Gat1. Regarding nitrate induction, calcineurin also affects the levels of transcription factors Gat2 and Yna2 involved in this process. We conclude that Ca(2+) and calcineurin play a central role in nitrogen signalling and assimilation. Thus, the nitrogen source modulates Ca(2+) content and calcineurin activation. Calcineurin in turn regulates nitrogen assimilation genes.

Bifunctional Au@Pt/Au nanoparticles as electrochemiluminescence signaling probes for SARS-CoV-2 detection.

A novel immunosensor based on electrochemiluminescence resonance energy transfer (ECL-RET) for the sensitive determination of N protein of the SARS-CoV-2 coronavirus is described. For this purpose, bifunctional core@shell nanoparticles composed of a Pt-coated Au core and finally decorated with small Au inlays (Au@Pt/Au NPs) have been synthesized to act as ECL acceptor, using [Ru (bpy)3]2+ as ECL donor. These nanoparticles are efficient signaling probes in the immunosensor developed. The proposed ECL-RET immunosensor has a wide linear response to the concentration of N protein of the SARS-CoV-2 coronavirus with a detection limit of 1.27 pg/mL. Moreover, it has a high stability and shows no response to other proteins related to different virus. The immunosensor has achieved the quantification of N protein of the SARS-CoV-2 coronavirus in saliva samples. Results are consistent with those provided by a commercial colorimetric ELISA kit. Therefore, the developed immunosensor provides a feasible and reliable tool for early and effective detection of the virus to protect the population.

Npr1 Ser/Thr protein kinase links nitrogen source quality and carbon availability with the yeast nitrate transporter (Ynt1) levels.

Ynt1, the single high affinity nitrate and nitrite transporter of the yeast Hansenula polymorpha, is regulated by the quality of nitrogen sources. Preferred nitrogen sources cause Ynt1 dephosphorylation, ubiquitinylation, endocytosis, and vacuolar degradation. In contrast, under nitrogen limitation Ynt1 is phosphorylated and sorted to the plasma membrane. We show here the involvement of the Ser/Thr kinase HpNpr1 in Ynt1 phosphorylation and regulation of Ynt1 levels in response to nitrogen source quality and the availability of carbon. In Δnpr1, Ynt1 phosphorylation does not take place, although Ynt1 ubiquitin conjugates increase. As a result, in this strain Ynt1 is sorted to the vacuole, from both plasma membrane and the later biosynthetic pathway in nitrogen-free conditions and nitrate. In contrast, overexpression of NPR1 blocks down-regulation of Ynt1, increasing Ynt1 phosphorylation at Ser-244 and -246 and reducing ubiquitinylation. Furthermore, Npr1 is phosphorylated in response to the preferred nitrogen sources, and indeed it is dephosphorylated in nitrogen-free medium. Under conditions where Npr1 is phosphorylated, Ynt1 is not and vice versa. We show for the first time that carbon starvation leads to Npr1 phosphorylation, whereas Ynt1 is dephosphorylated and degraded in the vacuole. Rapamycin prevents this, indicating a possible role of the target of rapamycin signaling pathway in this process. We concluded that Npr1 plays a key role in adapting Ynt1 levels to the nitrogen quality and availability of a source of carbon.

Visual attraction of the European tarnished plant bug Lygus rugulipennis (Hemiptera: Miridae) to a water trap with LED light in chrysanthemum greenhouses and olfactory attraction to novel compounds in Y-tube tests.

BACKGROUND: The European tarnished plant bug Lygus rugulipennis Poppius (Hemiptera: Miridae) can cause several types of damage to crops grown in greenhouses and fields, including flower abortion in eggplant, stem and fruit damage in cucumbers, and splits in chrysanthemums. Studies suggest that both male and female L. rugulipennis may be more attracted to traps based on visual attraction than pheromone-based trap. Therefore, the aim of this study was to evaluate the effectiveness of a water trap with LED lights and semiochemicals in trapping L. rugulipennis in the laboratory and greenhouse. RESULTS: The results showed that water traps equipped with white LED light caught 20 - 30 times more bugs than did the sex pheromone-based traps in greenhouse experiment. During the week of peak flight, the LED water trap caught a total of 29 males and females, whereas the sex pheromone caught only one male. Among the semiochemicals tested in a Y-tube, both males and females were attracted to ß-caryophyllene, but not in the presence of the sex pheromone, whereas both males and females were attracted to pentyl butyrate in the presence of the sex pheromone. The pheromone plus bean plant odor was attractive to the insects, suggesting an interaction between plant odor and pheromone. CONCLUSION: Overall, the findings of the study showed that the water trap with LED light could be an effective method for trapping L. rugulipennis in greenhouses. © 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Spectrum of Syphilitic Chorioretinitis and Its Evolution Based on Multimodal Imaging.

PURPOSE: To describe the different phenotypes of syphilitic chorioretinitis (SCR) and its clinical features in multimodal imaging (MI). METHODS: Complete ophthalmological examination and MI including spectral domain optic coherence tomography (SD-OCT), fundus autofluorescence (FAF) and fluorescein angiography (FA) were performed at diagnosis and during follow-up in patients diagnosed with SCR. RESULTS: 17 eyes of 10 patients were included. Clinical phenotypes included acute syphilitic posterior placoid chorioretinopathy (ASPPC), syphilitic outer retinitis (SOR) and punctate inner retinitis (PIR). Disruption of outer retinal layers were visible in all patients in SD-OCT. Inner retinal layers were only affected in PIR. FAF revealed macular hyperautofluorescence in all patients. FA showed maintained hyperfluorescence in ASPPC and SOR. CONCLUSIONS: ASPPC, SOR and PIR are different phenotypes of syphilitic chorioretinitis and present common features in SD-OCT and FAF. MI provides useful information to help the clinician during diagnosis and follow-up of these entities.

Influence of Gamification and Cooperative Work in Peer, Mixed and Interdisciplinary Teams on Emotional Intelligence, Learning Strategies and Life Goals That Motivate University Students to Study.

It is necessary to motivate university students to reduce the dropout rate in Spain, and to look for strategies that help university students acquire professional competencies; this is where gamification can be useful. The purpose of the study was to evaluate the influence of a learning methodology based on gamification and cooperative work in peers, and in mixed and interdisciplinary teams on the emotional intelligence, learning strategies, and life goals that motivate university students to learn. The sample consisted of 102 students who took a subject with a gamification-based methodology, through the Mechanics-Dynamics-Aesthetics model, in a cooperative way. The Self-Perception Scale of Personal Academic Motivation and the Trait Meta Mood Scale 24 were used. The results of the study showed an increase in learning strategies and life goals that motivated university students to study, as well as increases in emotional clarity and significantly in emotional repair. It was concluded that gamification is a positive tool for its ability to increase emotional intelligence, life goals, and learning strategies in university students' motivation to learn.

Enhancing the electrocatalytic activity of palladium nanocluster tags by selective introduction of gold atoms: Application for a wound infection biomarker detection.

In this work, an unprecedented study exploring the role that slight changes into the Pd/Au proportion have in the electrocatalytic activity of bimetallic Pd-AuNPs toward the oxygen reduction reaction (ORR) is conducted. In particular, a careful control of the amount of Au atoms introduced in the cluster and the evaluation of the optimum Pd:Au ratio for getting the maximum catalytic activity is performed for the first time. First, PdNPs are synthesized by alcohol reduction in the presence of polyvinylpyrrolidone, and gold atoms are selectively introduced on vertex or corner positions of the cluster in different amounts following a galvanic substitution procedure. Average elemental analysis done relying on EDX spectroscopy allows to evaluate the Pd:Au ratio in the Pd-AuNPs obtained. Lineal sweep voltammetry and chronoamperometry are used for the evaluation of the Pd-AuNPs electrocatalytic activity toward ORR at a neutral pH compared to PdNPs and AuNPs alone. Our results indicate that, the synergy between both metals is strongly enhanced when the amount of gold is controlled and occupies the more reactive positions of the cluster, reaching a maximum activity for the NPs containing a 30% of gold, while an excess of this metal leads to a decrease in such activity, as a shelter of the PdNPs is achieved. Chronoamperometric analysis allows the quantification of the optimal Pd-AuNPs at over 6 × 109 NPs/mL levels. Such optimal Pd-AuNPs were used as tags, taking advantage of the bio-functionalities of gold present in the cluster, in a proof-of-concept electrochemical immunosensor for the detection of hyaluronidase wound infection biomarker, using magnetic beads as platforms. Hyaluronidase was detected at levels as low as 50 ng/mL (0.02 U/mL; 437 U/mg) with good reproducibility (RSD below 8%) and selectivity (evaluated against bovine serum albumin, immunoglobulin G and lysozyme). The low matrix effects inherent to the use of magnetic bead platforms allowed us to discriminate between wound exudates with both sterile and infected ulcers without sample pre-treatment. This novel electrocatalytic immunoassay has the advantage, over common methods for NP tags electrochemical detection, of the signal generation in the same neutral medium where the immunoassay takes place (10 mM PBS pH 7.4), avoiding the use of additional and hazardous reagents, bringing it closer to their use as point-of-care devices. Overall, our findings may be of great interest not only for biosensing, but also for applications such as energy converting on fuel cells, in which the ORR has a pivotal role.

Electrical monitoring of infection biomarkers in chronic wounds using nanochannels.

Chronic wounds represent an important healthcare challenge in developed countries, being wound infection a serious complication with significant impact on patients' life conditions. However, there is a lack of methods allowing an early diagnosis of infection and a right decision making for a correct treatment. In this context, we propose a novel methodology for the electrical monitoring of infection biomarkers in chronic wound exudates, using nanoporous alumina membranes. Lysozyme, an enzyme produced by the human immune system indicating wound infection, is selected as a model compound to prove the concept. Peptidoglycan, a component of the bacterial layer and the native substrate of lysozyme, is immobilized on the inner walls of the nanochannels, blocking them both sterically and electrostatically. The steric blocking is dependent on the pore size (20-100 nm) and the peptidoglycan concentration, whereas the electrostatic blocking depends on the pH. The proposed analytical method is based on the electrical monitoring of the steric/electrostatic nanochannels unblocking upon the specific degradation of peptidoglycan by lysozyme, allowing to detect the infection biomarker at 280 ng/mL levels, which are below those expected in wounds. The low protein adsorption rate and thus outstanding filtering properties of the nanoporous alumina membranes allowed us to discriminate wound exudates from patients with both sterile and infected ulcers without any sample pre-treatment usually indispensable in most diagnostic devices for analysis of physiological fluids. Although size and charge effects in nanochannels have been previously approached for biosensing purposes, as far as we know, the use of nanoporous membranes for monitoring enzymatic cleavage processes, leading to analytical systems for the specific detection of the enzymes has not been deeply explored so far. Compared with previously reported methods, our methodology presents the advantages of no need of neither bioreceptors (antibodies or aptamers) nor competitive assays, low matrix effects and quantitative and rapid analysis at the point-of-care, being also of potential application for the determination of other protease biomarkers.

Ure2 is involved in nitrogen catabolite repression and salt tolerance via Ca2+ homeostasis and calcineurin activation in the yeast Hansenula polymorpha.

Disruption of HpURE2 resulted in a low expression of genes encoding nitrate-assimilatory proteins; sensitivity to Li(+), Na(+), and Cd(2+); no induction of ENA1; low levels of the GATA-type transcription factor Gat1; and low intracellular Ca(2+) levels. Gat1 levels were also very low in a Δcnb1 mutant lacking the regulatory subunit of calcineurin. The strain Δure2 was very sensitive to the calcineurin inhibitor FK506 and displayed several phenotypes reminiscent of Δcnb1. The reporter 4xCDRE-lacZ, containing calcineurin-dependent response elements in its promoter, revealed that calcineurin activation was reduced in HpΔure2. Expression of ScURE2 in Δure2 rescued nitrogen catabolite repression and Cd(2+) tolerance but not those phenotypes depending on calcineurin activation, such as salt tolerance and nitrate assimilation gene derepression. HpΔure2 showed an increased expression of the gene PMR1 encoding the Golgi Ca(2+)-ATPase, whereas that of PMC1 encoding the vacuolar Ca(2+)-ATPase remained unaltered. PMR1 up-regulation was abolished by deletion of the GATA-type transcription factor GAT2 in a HpΔure2 genetic background, and normal Ca(2+) levels were recovered. Moreover, overexpression of GAT2 or PMR1 yielded strains mimicking the phenotype of the HpΔure2. This suggests that the low Ca(2+) levels in the HpΔure2 mutant are due to the high levels of Pmr1 that replenish the Golgi Ca(2+) content, thus acting as a negative signal for Ca(2+) entry into the cell. We conclude that HpUre2 is involved in salt tolerance and also in nitrate assimilation gene derepression via Ca(2+) homeostasis regulation and calcineurin activation, which control the levels of Gat1.

Unfolded p53 as a Marker of Oxidative Stress in Mild Cognitive Impairment, Alzheimer's and Parkinson's Disease.

AIMS: There are several candidate biomarkers for AD and PD which differ in sensitivity, specificity, cost-effectiveness, invasiveness, logistical and technical demands. This study is aimed to test whether plasma concentration of unfolded p53 may help to discriminate among the neurodegenerative processes occurring in Mild Cognitive Impairment, Alzheimer's disease and Parkinson's disease. METHODS: An electrochemical immunosensor was used to measure unfolded p53 in plasma samples of 20 Mild Cognitive Impairment (13 males/7 females; mean age 74.95±5.31), 20 Alzheimer's (11 males/9 females; mean age: 77.25±7.79), 15 Parkinson's disease patients (12 males/3 females; mean age: 68.60 ± 7.36) and its respective age/sex/studies-matched controls. RESULTS: We observed a significantly higher concentration of unfolded p53 in the plasma of patients of each of the three pathologies with respect to their control groups (p=0.000). Furthermore, the plasma concentration of unfolded p53 was significantly higher in Alzheimer's disease patients in comparison with Mild Cognitive Impairment patients (p=0.000) and Parkinson's disease patients (p=0.006). No significant difference between Mild Cognitive Impairment and Parkinson's disease patients was observed (p=0.524). CONCLUSION: Our results suggest that unfolded p53 concentration in the plasma may be a useful biomarker for an undergoing neuropathological process that may be common, albeit with different intensity, to different diseases.

Emotions and Reactions to the Confinement by COVID-19 of Children and Adolescents With High Abilities and Community Samples: A Mixed Methods Research Study.

The goal of this research is to know and compare the emotions and reactions to confinement due to the COVID-19 pandemic in children and adolescents with high abilities and community samples. This is a mixed study with an exploratory reach that is descriptive, and which combines survey and qualitative methodologies to examine the emotions and reactions to confinement experiences of children and adolescents aged between 5 and 14 years. An online poll was designed with 46 questions, grouped into three sections: (1) General Data, (2) Reactions to the health contingencies of COVID-19, and (3) Positive and negative emotions. This last section was included the Oros' positive emotions questionnaire (2014) and the scales of the negative effect of the PANAS Schedule positive and negative affect on children and adolescents between 8 and 14 years old (Sandin, 2003). Data were collected online with a convenience sample. Quantitative data were analyzed with the SPSS statistics program and qualitative data with Alceste software. Among the main findings is the fact that there are no significant differences between the groups by ability in terms of reactions and emotions; however, there are differences between age groups and sex. The study discusses the implications for educational intervention in situations like the current pandemic.