mTert induction in p21-positive cells counteracts capillary rarefaction and pulmonary emphysema.

Lung diseases develop when telomeres shorten beyond a critical point. We constructed a mouse model in which the catalytic subunit of telomerase (mTert), or its catalytically inactive form (mTertCI), is expressed from the p21Cdkn1a locus. Expression of either TERT or TERTCI reduces global p21 levels in the lungs of aged mice, highlighting TERT non-canonical function. However, only TERT reduces accumulation of very short telomeres, oxidative damage, endothelial cell (ECs) senescence and senile emphysema in aged mice. Single-cell analysis of the lung reveals that p21 (and hence TERT) is expressed mainly in the capillary ECs. We report that a fraction of capillary ECs marked by CD34 and endowed with proliferative capacity declines drastically with age, and this is counteracted by TERT but not TERTCI. Consistently, only TERT counteracts decline of capillary density. Natural aging effects are confirmed using the experimental model of emphysema induced by VEGFR2 inhibition and chronic hypoxia. We conclude that catalytically active TERT prevents exhaustion of the putative CD34 + EC progenitors with age, thus protecting against capillary vessel loss and pulmonary emphysema.

Perceptions of COVID-19 patients in the use of bioethical principles and the physician-patient relationship: a qualitative approach.

BACKGROUND: The COVID-19 pandemic has influenced the approach to the health-disease system, raising the question about the principles of bioethics present in physician-patient relations. The principles while widely accepted may not be sufficient for a comprehensive ethical analysis. Therefore, the aim of this study was to explore the perception of these principles and the physician-patient relationship during a hospital stay through a qualitative approach. METHOD: Sixteen semi-structured interviews took place to know the patients' perception during their 2020 hospitalization for COVID-19. The data was analyzed through the constant comparison method, creating categories and comparing them. In the end, seven categories were established and were grouped in three: bioethical principles (dignity, charity, vulnerability, autonomy), doctor-patient relationship (participant commitment, informed consent, health staff-patient relationship) and the experience of the disease (illness, the role of the family). RESULTS: The research found that most patients described a positive experience, with the feeling of having been well cared for with no sense of discrimination or injustice done. The majority also reported that their autonomy was respected in the treatment decisions. The evaluation of these attitudes is an area of opportunity, especially when the patients' vulnerability is at risk. CONCLUSIONS: The ethics of virtue offers a better reflection of how human beings manifest themselves by emphasizing the development of virtuous character and behaviors that allow them to realize their values in life. Authorized by the Research Ethics Committee with registration: DI/18/105-B/3/308.

Changes in patients' religiosity related to dementia / Cambios en la religiosidad del paciente relacionados con la demencia.

Dementia is characterized by the presence of progressive cognitive and behavioral symptoms which affect normal functioning. With the purpose of determining if there are any changes related to patients' religiosity due to dementia, we developed a questionnaire aiming to evaluate changes pre and post diagnostic. In effect, we observed that patients with dementia might experience a reduction of the importance given to religion and its associated practices with the disease progression. However, God's belief did not show any changes despite the diagnosis. Therefore, we point out the relevance of incorporating religiosity as another aspect to take into account in the cognitive rehabilitation treatments.

La demencia se caracteriza por un inicio gradual y un deterioro cognitivo y conductual progresivo, que provoca un significativo impacto en el funcionamiento normal del paciente. Con el objetivo de determinar si hay cambios en la religiosidad de los pacientes con demencia, desarrollamos un cuestionario con el propósito de evaluar cambios en aspectos relacionados con la religiosidad de los pacientes 10 años atrás y en la actualidad. Este formulario fue contestado por familiares y cuidadores. El estudio mostró que los pacientes con demencia podrían experimentar una reducción de la importancia dada a la religión y a sus prácticas asociadas durante el curso de la enfermedad. Sin embargo, la creencia en Dios no se vería modificada a pesar del diagnóstico. Por lo tanto, y dado que se ha visto que la religiosidad puede ayudar en pacientes con demencia, señalamos la relevancia de tener en cuenta dicha variable y de incorporar estrategias para los tratamientos de rehabilitación cognitiva y para la psicoeducación del entorno del paciente.

Telomerase reverse transcriptase delays aging in cancer-resistant mice.

Telomerase confers limitless proliferative potential to most human cells through its ability to elongate telomeres, the natural ends of chromosomes, which otherwise would undergo progressive attrition and eventually compromise cell viability. However, the role of telomerase in organismal aging has remained unaddressed, in part because of the cancer-promoting activity of telomerase. To circumvent this problem, we have constitutively expressed telomerase reverse transcriptase (TERT), one of the components of telomerase, in mice engineered to be cancer resistant by means of enhanced expression of the tumor suppressors p53, p16, and p19ARF. In this context, TERT overexpression improves the fitness of epithelial barriers, particularly the skin and the intestine, and produces a systemic delay in aging accompanied by extension of the median life span. These results demonstrate that constitutive expression of Tert provides antiaging activity in the context of a mammalian organism.

Proteome Landscape during Ripening of Solid Endosperm from Two Different Coconut Cultivars Reveals Contrasting Carbohydrate and Fatty Acid Metabolic Pathway Modulation.

Cocos nucifera L. is a crop grown in the humid tropics. It is grouped into two classes of varieties: dwarf and tall; regardless of the variety, the endosperm of the coconut accumulates carbohydrates in the early stages of maturation and fatty acids in the later stages, although the biochemical factors that determine such behavior remain unknown. We used tandem mass tagging with synchronous precursor selection (TMT-SPS-MS3) to analyze the proteomes of solid endosperms from Yucatan green dwarf (YGD) and Mexican pacific tall (MPT) coconut cultivars. The analysis was conducted at immature, intermediate, and mature development stages to better understand the regulation of carbohydrate and lipid metabolisms. Proteomic analyses showed 244 proteins in YGD and 347 in MPT; from these, 155 proteins were shared between both cultivars. Furthermore, the proteomes related to glycolysis, photosynthesis, and gluconeogenesis, and those associated with the biosynthesis and elongation of fatty acids, were up-accumulated in the solid endosperm of MPT, while in YGD, they were down-accumulated. These results support that carbohydrate and fatty acid metabolisms differ among the developmental stages of the solid endosperm and between the dwarf and tall cultivars. This is the first proteomics study comparing different stages of maturity in two contrasting coconut cultivars and may help in understanding the maturity process in other palms.

Moonlight-like proteins are actually cell wall components in Pseudocercospora fijiensis.

The fungal cell wall protects fungi against threats, both biotic and abiotic, and plays a role in pathogenicity by facilitating host adhesion, among other functions. Although carbohydrates (e.g. glucans, chitin) are the most abundant components, the fungal cell wall also harbors ionic proteins, proteins bound by disulfide bridges, alkali-extractable, SDS-extractable, and GPI-anchored proteins, among others; the latter consisting of suitable targets which can be used for fungal pathogen control. Pseudocercospora fijiensis is the causal agent of black Sigatoka disease, the principal threat to banana and plantain worldwide. Here, we report the isolation of the cell wall of this pathogen, followed by extensive washing to eliminate all loosely associated proteins and conserve those integrated to its cell wall. In the HF-pyridine protein fraction, one of the most abundant protein bands was recovered from SDS-PAGE gels, electro-eluted and sequenced. Seven proteins were identified from this band, none of which were GPI-anchored proteins. Instead, atypical (moonlight-like) cell wall proteins were identified, suggesting a new class of atypical proteins, bound to the cell wall by unknown linkages. Western blot and histological analyses of the cell wall fractions support that these proteins are true cell wall proteins, most likely involved in fungal pathogenesis/virulence, since they were found conserved in many fungal pathogens.

Quality of life, mood, and cognitive performance in older adults with cognitive impairment during the first wave of COVID 19 in Argentina.

BACKGROUND AND OBJECTIVES: In Argentina, government has established lockdown on 19 March 2020 to decrease SARS-COV-2 infection. The study aim was to understand how mandatory quarantine imposed due to COVID-19 pandemic has affected quality of life, mood, and cognitive performance of older adults with cognitive impairment. DESIGN: Longitudinal descriptive-observational study. PARTICIPANTS: Patients with cognitive impairment attending to online cognitive training sessions. MEASUREMENTS: Participants have completed by themselves Quality of Life in Alzheimer's Disease scale (QOL-AD), Beck Depression Inventory (BDI-II), Test your Memory (TYM), and an attention and executive task created by our institution. Same assessments were done at the beginning of the lockdown and 7 months later. RESULTS: Fifty-one adults were included. An increase in BDI-II score (p = 0.049) and worse performance in one of the executive attention tests (p = 0.012) have been found. No significant differences in QOL-AD, TYM, verbal fluency, or memory tests total scores have been observed. Reviewing scales subitems, differences in changes in sleep habits (p = 0.021), energy level decrease (p = 0.004), worse subjective record of memory capacity (p = 0.028), and decrease in ability to do housework (p = 0.007) have been shown. In those who lived alone, BDI and TYM higher scores in BDI-II (p = 0.030) (p = 0.022) have been found. CONCLUSION: Mandatory quarantine imposed due to COVID-19 pandemic was associated with worsening of mood, some quality-of-life variables, and decrease in attention in older adults with cognitive impairment in Argentina.

Mitochondrial and nuclear DNA matching shapes metabolism and healthy ageing.

Human mitochondrial DNA (mtDNA) shows extensive within population sequence variability. Many studies suggest that mtDNA variants may be associated with ageing or diseases, although mechanistic evidence at the molecular level is lacking. Mitochondrial replacement has the potential to prevent transmission of disease-causing oocyte mtDNA. However, extension of this technology requires a comprehensive understanding of the physiological relevance of mtDNA sequence variability and its match with the nuclear-encoded mitochondrial genes. Studies in conplastic animals allow comparison of individuals with the same nuclear genome but different mtDNA variants, and have provided both supporting and refuting evidence that mtDNA variation influences organismal physiology. However, most of these studies did not confirm the conplastic status, focused on younger animals, and did not investigate the full range of physiological and phenotypic variability likely to be influenced by mitochondria. Here we systematically characterized conplastic mice throughout their lifespan using transcriptomic, proteomic,metabolomic, biochemical, physiological and phenotyping studies. We show that mtDNA haplotype profoundly influences mitochondrial proteostasis and reactive oxygen species generation,insulin signalling, obesity, and ageing parameters including telomere shortening and mitochondrial dysfunction, resulting in profound differences in health longevity between conplastic strains.

Fungal Effectoromics: A World in Constant Evolution.

Effectors are small, secreted molecules that mediate the establishment of interactions in nature. While some concepts of effector biology have stood the test of time, this area of study is ever-evolving as new effectors and associated characteristics are being revealed. In the present review, the different characteristics that underly effector classifications are discussed, contrasting past and present knowledge regarding these molecules to foster a more comprehensive understanding of effectors for the reader. Research gaps in effector identification and perspectives for effector application in plant disease management are also presented, with a focus on fungal effectors in the plant-microbe interaction and interactions beyond the plant host. In summary, the review provides an amenable yet thorough introduction to fungal effector biology, presenting noteworthy examples of effectors and effector studies that have shaped our present understanding of the field.

WideEffHunter: An Algorithm to Predict Canonical and Non-Canonical Effectors in Fungi and Oomycetes.

Newer effectorome prediction algorithms are considering effectors that may not comply with the canonical characteristics of small, secreted, cysteine-rich proteins. The use of effector-related motifs and domains is an emerging strategy for effector identification, but its use has been limited to individual species, whether oomycete or fungal, and certain domains and motifs have only been associated with one or the other. The use of these strategies is important for the identification of novel, non-canonical effectors (NCEs) which we have found to constitute approximately 90% of the effectoromes. We produced an algorithm in Bash called WideEffHunter that is founded on integrating three key characteristics: the presence of effector motifs, effector domains and homology to validated existing effectors. Interestingly, we found similar numbers of effectors with motifs and domains within two different taxonomic kingdoms: fungi and oomycetes, indicating that with respect to their effector content, the two organisms may be more similar than previously believed. WideEffHunter can identify the entire effectorome (non-canonical and canonical effectors) of oomycetes and fungi whether pathogenic or non-pathogenic, unifying effector prediction in these two kingdoms as well as the two different lifestyles. The elucidation of complete effectoromes is a crucial step towards advancing effectoromics and disease management in agriculture.

Identification of a Novel Lipase with AHSMG Pentapeptide in Hypocreales and Glomerellales Filamentous Fungi.

Lipases are enzymes that hydrolyze triglycerides to fatty acids and glycerol. A typical element in lipases is a conserved motif of five amino acids (the pentapeptide), most commonly G-X-S-X-G. Lipases with the pentapeptide A-X-S-X-G are present in species of Bacillus, Paucimonas lemoignei, and the yeast Trichosporon asahii; they are usually thermotolerant and solvent resistant. Recently, while searching for true lipases in the Trichoderma harzianum genome, one lipase containing the pentapeptide AHSMG was identified. In this study, we cloned from T. harzianum strain B13-1 the lipase ID135964, renamed here as ThaL, which is 97.65% identical with the reference. We found that ThaL is a lid-containing true lipase of cluster III that belongs to a large family comprising highly conserved proteins in filamentous fungi in the orders Hypocreales and Glomerellales, in which predominantly pathogenic fungi are found. ThaL was expressed in conidia, as well as in T. harzianum mycelium, where it was cultured in liquid minimal medium. These results-together with the amino acid composition, absence of a signal peptide, mitochondrial sorting prediction, disordered regions in the protein, and lineage-specific phylogenetic distribution of its homologs-suggest that ThaL is a non-canonical effector. In summary, AHSMG-lipase is a novel lipase family in filamentous fungi, and is probably involved in pathogenicity.

["ZOOM effect". The TV sign in patients with dementia: Is there an increased incidence during pandemia due to a rise in isolation and virtuality?] / "Efecto ZOOM". El signo de la TV en pacientes con demencia: ¿Hubo un aumento en la incidencia durante la pandemia debido al mayor aislamiento y la virtualidad?

Delusional misidentification syndromes are misperceptions of external stimuli with an associated belief or elaboration that is held with delusional intensity. In the TV sign patient believes that the observed television events are occurring in a real three-dimensional space. It is one of the very rare forms of delusional misidentification syndrome in patients withdementia.Wereport7patientswithcognitiveimpairmentcaseswhohavepresentedTVsignduringtheCOVID-19 pandemic. Two patients had Alzheimer's dementia type diagnosis, 1 atypical Alzheimer dementia, 1 vascular dementia and 3 of them had mixed etiology (2 Alzheimer dementia + vascular and 1 dementia with Lewy bodies + vascular). Three presented other psychotic symptoms and 1 patient also had Capgras syndrome. These 7 cases series raise the possibility of an increase incidence of TV sign in patients with dementia during pandemia triggered by the rise in expo- sure to screen devices and a social isolation during this period.

Do we treat dementia of the Alzheimer type or Alzheimer's disease? Anti-dementia drugs in the era of biomarkers / ¿Tratamos la demencia tipo Alzheimer o la enfermedad de Alzheimer? Fármacos antidemenciales en la era de los biomarcadores.

Approved drug treatments for Alzheimer´s disease (AD) are symptomatic and don´t modify the disease course. These include acetylcholinesterase inhibitors (AchI) and N-methyl-D-aspartate receptor antagonist, memantine. Around 20 years ago, these drugs were approved for Alzheimer type Dementia. This wasbased on clinical trials which inclusion criteria were focused on a clinical amnestic AD presentation. At that time, subjects with an atypical AD clinical presentation or biomarkers were not included in the pharmacological trials. New biomarkers that detect amyloid and neurodegeneration have allowed us to evaluate pathological changes compatible with AD. These new advances from aclinical and biomarkers perspective allowed a diagnostic criteria update; going from an exclusively clinical criteria to one that is hybrid: clinical presentation and biomarkers based criteria.New biomarkers facilitate the early diagnosis of AD and other dementias.However, they also generate new challenges and questions regarding the adequate pharmacological treatment.There is a need for clinical trials that evaluate anti-dementia drug's efficacy based on current diagnostic criteria (clinical profile and biomarkers) and new practice guidelines. In addition, regulatory authorities should update ACHI and memantine indications.This will help doctors to prescribe the best possible treatment for this specific population without increasing risks.

Los tratamientos farmacológicos aprobados para la enfermedad de Alzheimer (EA) son sintomáticos y no modifican el curso de la enfermedad. Estos incluyen inhibidores de la acetilcolinesterasa (IACE) y el antagonista del receptor de N-metil-D-aspartato, memantina. Estos medicamentos fueron aprobados para la demencia de tipo Alzheimer (DTA) hace unos 20 años, basándose en ensayos clínicos centrados en la presentación clínica amnésica de la EA sin considerar biomarcadores o presentaciones clínicas atípicas de EA. Los nuevos biomarcadores que detectan amiloide y neurodegeneración nos han permitido evaluar cambios patológicos compatibles con la EA. Estos nuevos avances desde la perspectiva de los biomarcadores y clínicos han llevado a una actualización de los criterios diagnósticos, pasando de criterios exclusivamente clínicos a criterios híbridos: clínicos y basados en marcadores. Estos biomarcadores facilitan el diagnóstico precoz de la EA y otras demencias; sin embargo, a veces generan desafíos y replanteos en relación al tratamiento farmacológico adecuado. Sería útil implementar ensayos clínicos que evalúen la eficacia de los fármacos aprobados para la enfermedad de Alzheimer, en su momento con criterios de demencia tipo Alzheimer en función de los criterios diagnósticos actuales (perfil clínico y biomarcadores). Además, la actualización de la indicación de prescripción de IACE y memantina por parte de las autoridades regulatorias especificando con más detalle la población objetivo ayudaría a prescribir el mejor tratamiento posible a los pacientes sin aumentar los riesgos.

Telomeres and telomerase in heart regeneration.

Although recent advances have overturned the old view of the human heart as an inert postmitotic organ, it is clear that the adult heart´s capacity to regenerate after an ischemic episode is very limited. Unlike humans, zebrafish and other lower vertebrates vigorously regenerate damaged myocardium after cardiac injury. Understanding how the zebrafish is able to conserve life-long cardiac regeneration capacity while mammals lose it soon after birth is crucial for the development of new treatments for myocardial infarction. Mammals and lower vertebrates differ markedly in their rates of cardiomyocyte proliferation and levels of telomerase activity. Here, we review recent discoveries identifying lack of telomerase activity and concomitant telomere dysfunction as natural barriers to cardiomyocyte proliferation and cardiac regeneration.

Early Detection of the Fungal Banana Black Sigatoka Pathogen Pseudocercospora fijiensis by an SPR Immunosensor Method.

Black Sigatoka is a disease that occurs in banana plantations worldwide. This disease is caused by the hemibiotrophic fungus Pseudocercospora fijiensis, whose infection results in a significant reduction in both product quality and yield. Therefore, detection and identification in the early stages of this pathogen in plants could help minimize losses, as well as prevent the spread of the disease to neighboring cultures. To achieve this, a highly sensitive SPR immunosensor was developed to detect P. fijiensis in real samples of leaf extracts in early stages of the disease. A polyclonal antibody (anti-HF1), produced against HF1 (cell wall protein of P. fijiensis) was covalently immobilized on a gold-coated chip via a mixed self-assembled monolayer (SAM) of alkanethiols using the EDC/NHS method. The analytical parameters of the biosensor were established, obtaining a limit of detection of 11.7 µg mL-1, a sensitivity of 0.0021 units of reflectance per ng mL-1 and a linear response range for the antigen from 39.1 to 122 µg mL-1. No matrix effects were observed during the measurements of real leaf banana extracts by the immunosensor. To the best of our knowledge, this is the first research into the development of an SPR biosensor for the detection of P. fijiensis, which demonstrates its potential as an alternative analytical tool for in-field monitoring of black Sigatoka disease.

The cell wall proteome from two strains of Pseudocercospora fijiensis with differences in virulence.

Pseudocercospora fijiensis causes black Sigatoka disease, the most important threat to banana. The cell wall is crucial for fungal biological processes, including pathogenesis. Here, we performed cell wall proteomics analyses of two P. fijiensis strains, the highly virulent Oz2b, and the less virulent C1233 strains. Strains were starved from nitrogen to mimic the host environment. Interestingly, in vitro cultures of the C1233 strain grew faster than Oz2b in PDB medium, suggesting that C1233 survives outside the host better than the highly virulent Oz2b strain. Both strains were submitted to nitrogen starvation and the cell wall proteins were isolated and subjected to nano-HPLC-MS/MS. A total of 2686 proteins were obtained from which only 240 had a known function and thus, bioinformatics analyses were performed on this group. We found that 90 cell wall proteins were shared by both strains, 21 were unique for Oz2b and 39 for C1233. Shared proteins comprised 24 pathogenicity factors, including Avr4 and Ecp6, two effectors from P. fijiensis, while the unique proteins comprised 16 virulence factors in C1233 and 11 in Oz2b. The P. fijiensis cell wall proteome comprised canonical proteins, but thirty percent were atypical, a feature which in other phytopathogens has been interpreted as contamination. However, a comparison with the identities of atypical proteins in other reports suggests that the P. fijiensis proteins we detected were not contaminants. This is the first proteomics analysis of the P. fijiensis cell wall and our results expands the understanding of the fundamental biology of fungal phytopathogens and will help to decipher the molecular mechanisms of pathogenesis and virulence in P. fijiensis.

[Head turn sign as a marker of dementia diagnosis]. / Signo del giro de la cabeza como marcador para el diagnóstico de demencia.

Due to the phenomenon of ageing population, cognitive impairment has become more prevalent, and any marker able to improve its detection becomes of significant importance. With that objective in mind, a prospective observational study was performed in a medical consultation related to cognitive impairment. 150 subjects (58% women) were included with an average age of 76 years (SD 8.6). The Head Turn sign was positive for 32 subjects with a sensitivity of 41.7% for dementia, a specificity of 93.7% and a negative predictive value of 64.4%. This study shows the semiologic value of a simple clinical sign, easy to spot in the daily practice and most helpful to alert physicians about a likely dementia diagnosis.

Telomere Length Defines the Cardiomyocyte Differentiation Potency of Mouse Induced Pluripotent Stem Cells.

Induced pluripotent stem cells (iPSCs) can be differentiated in vitro and in vivo to all cardiovascular lineages and are therefore a promising cell source for cardiac regenerative therapy. However, iPSC lines do not all differentiate into cardiomyocytes (CMs) with the same efficiency. Here, we show that telomerase-competent iPSCs with relatively long telomeres and high expression of the shelterin-complex protein TRF1 (iPSChighT ) differentiate sooner and more efficiently into CMs than those with relatively short telomeres and low TRF1 expression (iPSClowT ). Ascorbic acid, an enhancer of cardiomyocyte differentiation, further increases the cardiomyocyte yield from iPSChighT but does not rescue the cardiomyogenic potential of iPSClowT . Interestingly, although iPSCslowT differentiate very poorly to the mesoderm and endoderm lineages, they differentiate very efficiently to the ectoderm lineage, indicating that cell fate can be determined by in vitro selection of iPSCs with different telomere content. Our findings highlight the importance of selecting iPSCs with ample telomere reserves in order to generate high numbers of CMs in a fast, reliable, and efficient way. Stem Cells 2017;35:362-373.