A Missense Variant in TP53 Could Be a Genetic Biomarker Associated with Bone Tissue Alterations.

Metabolic bone diseases cover a broad spectrum of disorders that share alterations in bone metabolism that lead to a defective skeleton, which is associated with increasing morbidity, disability, and mortality. There is a close connection between the etiology of metabolic bone diseases and genetic factors, with TP53 being one of the genes associated therewith. The single nucleotide polymorphism (SNP) Arg72Pro of TP53 is a genetic factor associated with several pathologies, including cancer, stroke, and osteoporosis. Here, we aim to analyze the influence of the TP53 Arg72Pro SNP on bone mass in humanized Tp53 Arg72Pro knock-in mice. This work reports on the influence of the TP53 Arg72Pro polymorphism in bone microarchitecture, OPG expression, and apoptosis bone status. The results show that the proline variant of the TP53 Arg72Pro polymorphism (Pro72-p53) is associated with deteriorated bone tissue, lower OPG/RANK ratio, and lower apoptosis in bone tissue. In conclusion, the TP53 Arg72Pro polymorphism modulates bone microarchitecture and may be a genetic biomarker that can be used to identify individuals with an increased risk of suffering metabolic bone alterations.

Retinal Neuroprotective Effect of Mesenchymal Stem Cells Secretome Through Modulation of Oxidative Stress, Autophagy, and Programmed Cell Death.

Purpose: Degenerative mechanisms of retinal neurodegenerative diseases (RND) share common cellular and molecular signalization pathways. Curative treatment does not exist and cell-based therapy, through the paracrine properties of mesenchymal stem cells (MSC), is a potential unspecific treatment for RND. This study aimed to evaluate the neuroprotective capability of human bone marrow (bm) MSC secretome and its potential to modulate retinal responses to neurodegeneration. Methods: An in vitro model of spontaneous retinal neurodegeneration was used to compare three days of monocultured neuroretina (NR), NR cocultured with bmMSC, and NR cultured with bmMSC secretome. We evaluated retinal morphology markers (Lectin peanut agglutinin, rhodopsin, protein kinase C α isoform, neuronal-specific nuclear protein, glial fibrillary acidic protein, TdT-mediated dUTP nick-end labeling, and vimentin) and proteins involved in apoptosis (apoptosis-inductor factor, caspase-3), necroptosis (MLKL), and autophagy (p62). Besides, we analyzed the relative mRNA expression through qPCR of genes involved in apoptosis (BAX, BCL2, CASP3, CASP8, CASP9), necroptosis (MLKL, RIPK1, RIPK3), autophagy (ATG7, BCLIN1, LC3B, mTOR, SQSTM1), oxidative stress (COX2, CYBA, CYBB, GPX6, SOD1, TXN2, TXNRD1) and inflammation (IL1, IL6, IL10, TGFb1, TNFa). Results: The bmMSC secretome preserves retinal morphology, limits pro-apoptotic- and pro-necroptotic-related gene and protein expression, modulates autophagy-related genes and proteins, and stimulates the activation of antioxidant-associated genes. Conclusions: The neuroprotective ability of the bmMSC secretome is associated with activation of antioxidant machinery, modulation of autophagy, and inhibition of apoptosis and necroptosis during retinal degeneration. The neuroprotective effect of bmMSC secretomes in the presence/absence of MSC looks similar. Our current results reinforce the hypothesis that the human bmMSC secretome slows retinal neurodegeneration and may be a therapeutic option for treating RND.

Programmed Cell Death and Autophagy in an in vitro Model of Spontaneous Neuroretinal Degeneration.

Retinal neurodegenerative diseases are the leading causes of visual impairment and irreversible blindness worldwide. Although the retinal response to injury remains closely similar between different retinal neurodegenerative diseases, available therapeutic alternatives are only palliative, too expensive, or very specific, such as gene therapy. In that sense, the development of broad-spectrum neuroprotective therapies seems to be an excellent option. In this regard, it is essential to identify molecular targets involved in retinal degeneration, such as cell death mechanisms. Apoptosis has been considered as the primary cell death mechanism during retinal degeneration; however, recent studies have demonstrated that the only use of anti-apoptotic drugs is not enough to confer good neuroprotection in terms of cell viability and preservation. For that reason, the interrelationship that exists between apoptosis and other cell death mechanisms needs to be characterized deeply to design future therapeutic options that simultaneously block the main cell death pathways. In that sense, the study aimed to characterize the programmed cell death (in terms of apoptosis and necroptosis) and autophagy response and modulation in retinal neurodegenerative diseases, using an in vitro model of spontaneous retinal neurodegeneration. For that purpose, we measured the mRNA relative expression through qPCR of a selected pool of genes involved in apoptosis (BAX, BCL2, CASP3, CASP8, and CASP9), necroptosis (MLKL, RIPK1, and RIPK3), and autophagy (ATG7, BCLIN1, LC3B, mTOR, and SQSTM1); besides, the immunoexpression of their encoding proteins (Casp3, MLKL, RIPK1, LC3B, and p62) were analyzed using immunohistochemistry. Our results showed an increase of pro-apoptotic and pro-necroptotic related genes and proteins during in vitro retinal neurodegeneration. Besides, we describe for the first time the modulation between programmed cell death mechanisms and autophagy in an in vitro retinal neurodegeneration model. This study reinforces the idea that cell death mechanisms are closely interconnected and provides new information about molecular signaling and autophagy along the retinal degeneration process.

[Performance assessment in microscopic diagnosis of malaria in the laboratory network of the National Institute of Health of Peru, 2012-2017]. / Evaluación del desempeño en el diagnóstico microscópico de malaria en la red de laboratorios del Instituto Nacional de Salud De Perú, 2012-2017.

Timely diagnosis of malaria is a strategy proposed by the World Health Organization to reduce malaria morbidity and mortality. A study was conducted to assess performance in microscopic diagnosis of malaria in the network of laboratories under the National Malaria Reference Laboratory of the National Institute of Health between 2012 and 2017. In the years of study, the laboratories obtained a rating of "acceptable" in the diagnosis of Plasmodium by 38.4%, 43.7%, 60.0%, 83.3%, 90.9%, and 95.8%, respectively, in the evaluation of species by 0%, 6.2%, 15.0%, 50.0%, 40.9%, and 54.1%, respectively; in stage assessment by 23.0%, 25.0%, 35.0%, 83.3%, 54.5%, and 79.1%, respectively; and in parasitic density assessment by 0%, 6.2%, 10.0%, 33.3%, 0%, and 12.5%, respectively. We conclude that in the period under evaluation, the percentage of laboratories that diagnosed and recognized the species and stage increases, which is not the case for recognition of parasitic density.

El diagnóstico oportuno de malaria es una estrategia propuesta por la Organización Mundial de la Salud para reducir la morbimortalidad por esta enfermedad. Se realizó un estudio con el objetivo de evaluar el desempeño en el diagnóstico microscópico de malaria en la red de laboratorios pertenecientes al Laboratorio de Referencia Nacional de Malaria del Instituto Nacional de Salud entre 2012 y 2017. En los años de estudio, los laboratorios tuvieron la calificación de aceptable en el diagnóstico de Plasmodium en un 38,4%, 43,7%, 60,0%, 83,3%, 90,9% y 95,8%, respectivamente; en la evaluación de especie en un 0%, 6,2%, 15,0%, 50,0%, 40,9% y 54,1%, respectivamente; en la evaluación de estadio en un 23,0%, 25,0%, 35,0%, 83,3%, 54,5% y 79,1%, respectivamente; y en la evaluación de densidad parasitaria en un 0%, 6,2%, 10,0%, 33,3%, 0% y 12,5%, respectivamente. Concluimos que en el periodo evaluado se incrementó el porcentaje de laboratorios que diagnosticaron, reconocieron la especie y estadio, mas no el reconocimiento de densidad parasitaria.

Evaluación del desempeño en el diagnóstico microscópico de malaria en la red de laboratorios del Instituto Nacional de Salud De Perú, 2012-2017 / Performance assessment in microscopic diagnosis of malaria in the laboratory network of the National Institute of Health of Peru, 2012-2017

RESUMEN El diagnóstico oportuno de malaria es una estrategia propuesta por la Organización Mundial de la Salud para reducir la morbimortalidad por esta enfermedad. Se realizó un estudio con el objetivo de evaluar el desempeño en el diagnóstico microscópico de malaria en la red de laboratorios pertenecientes al Laboratorio de Referencia Nacional de Malaria del Instituto Nacional de Salud entre 2012 y 2017. En los años de estudio, los laboratorios tuvieron la calificación de aceptable en el diagnóstico de Plasmodium en un 38,4%, 43,7%, 60,0%, 83,3%, 90,9% y 95,8%, respectivamente; en la evaluación de especie en un 0%, 6,2%, 15,0%, 50,0%, 40,9% y 54,1%, respectivamente; en la evaluación de estadio en un 23,0%, 25,0%, 35,0%, 83,3%, 54,5% y 79,1%, respectivamente; y en la evaluación de densidad parasitaria en un 0%, 6,2%, 10,0%, 33,3%, 0% y 12,5%, respectivamente. Concluimos que en el periodo evaluado se incrementó el porcentaje de laboratorios que diagnosticaron, reconocieron la especie y estadio, mas no el reconocimiento de densidad parasitaria.

ABSTRACT Timely diagnosis of malaria is a strategy proposed by the World Health Organization to reduce malaria morbidity and mortality. A study was conducted to assess performance in microscopic diagnosis of malaria in the network of laboratories under the National Malaria Reference Laboratory of the National Institute of Health between 2012 and 2017. In the years of study, the laboratories obtained a rating of "acceptable" in the diagnosis of Plasmodium by 38.4%, 43.7%, 60.0%, 83.3%, 90.9%, and 95.8%, respectively, in the evaluation of species by 0%, 6.2%, 15.0%, 50.0%, 40.9%, and 54.1%, respectively; in stage assessment by 23.0%, 25.0%, 35.0%, 83.3%, 54.5%, and 79.1%, respectively; and in parasitic density assessment by 0%, 6.2%, 10.0%, 33.3%, 0%, and 12.5%, respectively. We conclude that in the period under evaluation, the percentage of laboratories that diagnosed and recognized the species and stage increases, which is not the case for recognition of parasitic density.