The immunity and redox clocks in mice, markers of lifespan.

Immune function and redox markers are used for estimating the aging rate, namely biological age (BA). However, it is unknown if this BA and its changes can be reflected in longevity. Thus, we must quantify BA in experimental animals. In peritoneal immune cells of 202 female mice (ICR/CD1) in different ages, 10 immune and 6 redox parameters were evaluated to construct two mathematical models for BA quantification in mice by multiple linear regression. Immune and redox parameters were selected as independent variables and chronological age as dependent, developing two models: the Immunity and the Redox Clocks, reaching both an adjusted R2 of 80.9% and a standard error of 6.38 and 8.57 weeks, respectively. Both models were validated in a different group of healthy mice obtaining a Pearson's correlation coefficient of 0.844 and 0.800 (p < 0.001) between chronological and BA. Furthermore, they were applied to adult prematurely aging mice, which showed a higher BA than non-prematurely aging mice. Moreover, after positive and negative lifestyle interventions, mice showed a lower and higher BA, respectively, than their age-matched controls. In conclusion, the Immunity and Redox Clocks allow BA quantification in mice and both the ImmunolAge and RedoxAge in mice relate to lifespan.

Development of novel antimicrobial acrylic denture modified with copper nanoparticles.

PURPOSE: This study aimed to synthesize heat-cured poly(methyl methacrylate) (PMMA) acrylic formulated with copper nanoparticles (nCu) for producing dentures with antimicrobial properties and ability to prevent denture stomatitis (DS). METHODS: nCu/PMMA nanocomposites were prepared through in situ formation of nCu into methyl methacrylate (MMA). The fabricated material was characterized using scanning electron microscopy, spectroscopy (energy-dispersive X-ray, attenuated total reflectance-Fourier-transform infrared, and X-ray photoelectron spectroscopy), X-ray diffraction analysis, and mechanical flexural tests (ISO 20795-1:2008). Antimicrobial activity against Candida albicans and oral bacteria was determined. MTS assay (ISO 10993-5:2009) and copper release experiments were conducted to assess cytotoxicity. In the clinical trial, participants wearing nCu/PMMA (n=25) and PMMA (n=25) dentures were compared; specifically, DS incidence and severity and Candida species proliferation were assessed for 12 months. Data were analyzed using analysis of variance with Tukey's post hoc test (α=0.05). RESULTS: nCu/PMMA nanocomposite loaded with 0.045% nCu exhibited the maximum antimicrobial activity against C. albicans and other oral bacteria without producing cytotoxicity in the wearer. nCu/PMMA dentures retained their mechanical and aesthetic properties as well as inhibited the growth of Candida species on both denture surface and patient palate. DS incidence and severity were lower in the nCu/PMMA denture group than in the PMMA denture group. CONCLUSIONS: PMMA acrylic produced with copper nanotechnology is antimicrobial, biocompatible, and aesthetic and can reduce DS incidence. Thus, this material may act as a novel preventive alternative for oral infections associated with denture use.

Positive effects of pulsed electromagnetic fields on behavior, immune function, and oxidative and inflammatory state in old mice.

The establishment of chronic oxidative and inflammatory stress with aging leads to the deterioration of the nervous and immune systems and, consequently, to the loss of health. The aim of this work was to study the effect of exposure to low-frequency pulsed electromagnetic fields (PEMFs) produced by the NEURALTER® system (15 min/day for 4 weeks) in the behavior, immune functions, and oxidative and inflammatory state of old mice. Female old CD1 mice were divided into three groups: control group, handling control group and Neuralter group. Then, behavioral tests were performed, and peritoneal leukocytes were extracted to analyze function, oxidative and inflammatory parameters. In peritoneal leukocytes from old mice, the effects in vitro of 15 min with NEURALTER® were studied on function and oxidative parameters. The results show that after this type of treatment, old mice had greater coordination and locomotion, better immune function, and an oxidative-inflammatory state. Similarly, the immune function and oxidative state of leukocytes showed an improvement when these cells were exposed directly to the NEURALTER® system. In conclusion, the exposure to low-frequency PEMFs produced by the NEURALTER® system has beneficial effects on health in aging. In addition, this effect is direct, at least in part, on immune cells.

Components of the Glutathione Cycle as Markers of Biological Age: An Approach to Clinical Application in Aging.

The oxidative-inflammatory theory of aging states that aging is the result of the establishment of a chronic oxidative-inflammatory stress situation in which the immune system is implicated. Among the redox parameters, those involved in the glutathione cycle have been suggested as essential in aging. Thus, the first objective of this study was to determine if several components of the glutathione cycle (glutathione reductase (GR) and glutathione peroxidase (GPx) activities, and concentrations of oxidized glutathione (GSSG) and reduced glutathione (GSH)) in leukocytes) are associated with the biological age (ImmunolAge) estimated using the Immunity Clock in 190 men and women. The second objective was to identify the best blood fraction (whole blood, blood cells, erythrocytes, or plasma) to quantify these components and correlate them with the estimated ImmunolAge. The results show that the oxidative state of peripheral leukocytes correlates with their functionality, supporting the idea that this is the basis of immunosenescence. In blood, the correlations are more significant in the fraction of blood cells with respect to ImmunolAge (positive correlations with GSSG concentration and the GSSG/GSH ratio, and negative correlations with GPx and GR activities). Therefore, blood cells are proposed as the most effective sample to estimate the biological age of individuals in clinical settings.

In Response to a Punctual Stress Male and Female Tyrosine Hydroxylase Haploinsufficient Mice Show a Deteriorated Behavior, Immunity, and Redox State.

An inadequate stress response is associated with impaired neuroimmunoendocrine communication, increasing morbidity and mortality. Since catecholamines (CA) constitute one of the acute stress response pathways, female mice with an haploinsufficiency of the tyrosine hydroxylase gene (TH-HZ), the main limiting enzyme in CA synthesis, show low CA amounts, exhibiting an impairment of homeostatic systems. The aim of this study was to investigate the effect of a punctual stress in TH-HZ mice, determining the differences with wild-type (WT) mice and those due to sex by restraint with a clamp for 10 min. After restraint, a behavioral battery was performed, and several immune functions, redox state parameters, and CA amounts were evaluated in peritoneal leukocytes. Results show that this punctual stress impaired WT behavior and improved female WT immunity and oxidative stress, whereas in TH-HZ mice, all parameters were impaired. In addition, different responses to stress due to sex were observed, with males having a worse response. In conclusion, this study confirms that a correct CA synthesis is necessary to deal with stress, and that when a positive stress (eustress) occurs, individuals may improve their immune function and oxidative state. Furthermore, it shows that the response to the same stressor is different according to sex.

[Touch, a crucial sense in social interactions to improve homeostasis in aging and promote healthy longevity]. / El tacto, un sentido crucial en las interacciones sociales para mejorar la homeostasis en el envejecimiento y promover una longevidad saludable.

Aging is associated with the generalized deterioration of the organism, being of great relevance experienced by homeostatic systems such as the nervous, immune, and endocrine systems, which increases the risk of morbidity and mortality. Among the lifestyle strategies that have been researched to improve these systems and achieve greater healthy longevity, this review will focus on the social environment. In order to verify the effectiveness of these both in the improvement of homeostasis and in life expectancy, the research carried out with experimental animals that have allowed this to be done will be discussed. In addition, as it has been observed that physical contact is crucial for the positive outcomes of social interaction on homeostatic systems and longevity to occur, we will focus on that mechanism, as well as some of the possible molecular pathways underlying the effects found.

Plasma acylcarnitines and gut-derived aromatic amino acids as sex-specific hub metabolites of the human aging metabolome.

Aging biology entails a cell/tissue deregulated metabolism that affects all levels of biological organization. Therefore, the application of "omic" techniques that are closer to phenotype, such as metabolomics, to the study of the aging process should be a turning point in the definition of cellular processes involved. The main objective of the present study was to describe the changes in plasma metabolome associated with biological aging and the role of sex in the metabolic regulation during aging. A high-throughput untargeted metabolomic analysis was applied in plasma samples to detect hub metabolites and biomarkers of aging incorporating a sex/gender perspective. A cohort of 1030 healthy human adults (45.9% females, and 54.1% males) from 50 to 98 years of age was used. Results were validated using two independent cohorts (1: n = 146, 53% females, 30-100 years old; 2: n = 68, 70% females, 19-107 years old). Metabolites related to lipid and aromatic amino acid (AAA) metabolisms arose as the main metabolic pathways affected by age, with a high influence of sex. Globally, we describe changes in bioenergetic pathways that point to a decrease in mitochondrial ß-oxidation and an accumulation of unsaturated fatty acids and acylcarnitines that could be responsible for the increment of oxidative damage and inflammation characteristic of this physiological process. Furthermore, we describe for the first time the importance of gut-derived AAA catabolites in the aging process describing novel biomarkers that could contribute to better understand this physiological process but also age-related diseases.

Sex differences in neuroimmunoendocrine communication. Involvement on longevity.

Endocrine, nervous, and immune systems work coordinately to maintain the global homeostasis of the organism. They show sex differences in their functions that, in turn, contribute to sex differences beyond reproductive function. Females display a better control of the energetic metabolism and improved neuroprotection and have more antioxidant defenses and a better inflammatory status than males, which is associated with a more robust immune response than that of males. These differences are present from the early stages of life, being more relevant in adulthood and influencing the aging trajectory in each sex and may contribute to the different life lifespan between sexes.

Skin-to-Skin Contact: Crucial for Improving Behavior, Immunity, and Redox State after Short Cohabitation of Chronologically Old Mice and Prematurely Aging Mice with Adult Mice.

(1) Background: Aging is characterized by a deterioration of the homeostatic systems, namely the nervous and immune systems. The rate of aging can be modified by lifestyle factors such as social interactions. Recently, improvements in behavior, immune function, and oxidative state were observed in adult prematurely aging mice (PAM) and chronologically old mice after cohabitation with exceptional non-PAM (E-NPAM) and adult mice, respectively, for 2 months. However, the cause of this positive effect is not known. The objective of the present work was to study whether skin-to-skin contact promotes these improvements both in chronologically old mice and in adult PAM. (2) Methods: Old and adult CD1 female mice were used as well as adult PAM and E-NPAM. After cohabitation for 15 min/day for 2 months (two old mice or PAM with five adult mice or E-NPAM, respectively, with both non- and skin-to-skin contact), several behavioral tests were performed and functions and oxidative stress parameters in peritoneal leukocytes were analyzed. (3) Results: This social interaction improved behavioral responses, immune functions, redox state, and longevity, but only if the animals had skin-to-skin contact. (4) Conclusions: Physical contact seems to be crucial to experiencing the positive effects of social interaction.

Complicaciones de la técnica anestésica mandibular Spix en la clínica odontológica de la Universidad Andrés Bello / Incidence of complications of mandibular anesthetic Spix techniques in the Andrés Bello University dental clinic

Objetivo: Determinar la incidencia de complicaciones de la técnica anestésica Spix en procedimientos odontológicos a pacientes atendidos en la clínica de la Universidad Andrés Bello (UNAB). Material y Métodos: Se analizó a 37 pacientes que fueron atendidos por alumnos de cuarto y quinto año de la clínica odontológica, a los cuales se le realizó la técnica anestésica Spix para realizar el procedimiento odontológico. Se consignó mediante la observación la presencia de formación de hematomas intraorales en el sitio de punción, rotura de la aguja, cantidad de tubos de solución anestésica inyectados, presencia de dolor a la inyección de solución anestésico y la presencia o no de parálisis facial. Mediante la recolección de datos y posterior encuesta a los participantes se consignó la presencia de trismus al día siguiente de la atención y parestesia persistente al día siguiente de la atención. Resultados: De 37 pacientes estudiados que recibieron la técnica anestésica Spix, 6 presentaron hematoma intraoral (16,2%), ninguno reportó rotura de la aguja, 1 presentó parálisis facial (2,7%), 1 presentó parestesia persistente al día siguiente (2,7%), 12 presentaron trismus posterior a la inyección (32,4%). El rango de dolor reportado fue entre 1 y 4 según la escala EVA. Conclusión: Hay una baja incidencia de las complicaciones asociadas a la técnica anestésica Spix en la clínica odontológica de la UNAB, siendo el trismus la complicación más frecuente. Se necesita un mayor número de muestra para entender mejor esta realidad.

Objective: To determine the incidence of complications of the Spix anesthetic technique in the dental procedures of patients attended at the Andrés Bello University dental clinic. Material and Methods: 37 patients who were cared for by fourth- and fifth-year students from the dental clinic of the Andrés Bello University were analyzed, who underwent the Spix anesthetic technique to perform the dental procedure. The presence of intraoral hematoma formation at the puncture site, needle breakage, number of injected anesthesia tubes, presence of pain upon injection of anesthetic and the presence or not of facial paralysis were recorded. Through data collection and subsequent survey of the participants, the presence of trismus was recorded the day after care. Results. Of the 37 cases of patients studied who received the Spix anesthetic technique, 6 had intraoral hematoma (16.2%), no needle break was reported, 1 had facial paralysis (2.7%), 1 had persistent paresthesia at the next day (2.7%), 12 presented trismus after the injection (32.4%), the pain range was between 1 and 4 according to the VAS scale. Conclusion. There is a low incidence of complications associated with the Spix anesthetic technique in the Andrés Bello University dental clinic, trismus being the most frequent (32.4%). A larger sample number should be needed to better understand this reality.

[Improved immunity and mean lifespan in mature mice genetically deficient in catecholamine synthesis after living with wild type for two months]. / Mejoría de la inmunidad y la esperanza de vida en ratones maduros, genéticamente deficientes en la síntesis de catecolaminas, tras convivir con controles durante dos meses.

INTRODUCTION: Mice hemizygous in tyrosine hydroxylase (TH-HZ), the limiting enzyme in catecholamine synthesis, show premature immunosenescence, which in females is associated with a shorter lifespan than the corresponding controls (WT). The coexistence of TH-Hz with WT improves the immune function in both males and females in adulthood. OBJECTIVE: To test whether cohabitation for two months of mature male TH-HZ with WT improves the immune function of the former and whether this impacts the lifespan. MATERIAL AND METHODS: Mature male ICR-CD1 mice (13 ± 1 months) TH-HZ coexisted with WT (2:4 ratio in each cage) for two months. Peritoneal leukocytes were extracted from all animals at baseline, one month, and two months after cohabitation, and macrophage phagocytic capacity, macrophage and lymphocyte chemotaxis, natural killer (NK) antitumor activity, and lymphoproliferative capacity in response to the mitogens concanavalin A and lipopolysaccharide (LPS) were assessed. The animals were maintained under these conditions until their natural death. RESULTS: The TH-HZ, which start, in general, with lower values than the WT in the immune functions studied, improved them after two months of cohabitation, becoming similar to those of the controls. This improvement was already observed in NK activity after one month of cohabitation. The TH-HZ presented lower mean longevity than WT, but when they cohabited with WT, it was similar to the latter. CONCLUSION: The coexistence of TH-HZ male mice with WT mice for two months at mature age improves these genetically modified animals' immune response and longevity.

Possibilities of using T-cell biophysical biomarkers of ageing.

Ageing is interrelated with the development of immunosenescence. This article focuses on one of the cell sets of the adaptive immune system, T cells, and provides a review of the known changes in T cells associated with ageing. Such fundamental changes affect both cell molecular content and internal ordering. However, acquiring a complete description of the changes at these levels would require extensive measurements of parameters and, furthermore, important fine details of the internal ordering that may be difficult to detect. Therefore, an alternative approach for the characterisation of cells consists of the performance of physical measurements of the whole cell, such as deformability measurements or migration measurements: the physical parameters, complementing the commonly used chemical biomarkers, may contribute to a better understanding of the evolution of T-cell states during ageing. Mechanical measurements, among other biophysical measurements, have the advantage of their relative simplicity: one single parameter agglutinates the complex effects of the variety of changes that gradually appear in cells during ageing.

Aging is accompanied by T-cell stiffening and reduced interstitial migration through dysfunctional nuclear organization.

Age-associated changes in T-cell function play a central role in immunosenescence. The role of aging in the decreased T-cell repertoire, primarily because of thymic involution, has been extensively studied. However, increasing evidence indicates that aging also modulates the mechanical properties of cells and the internal ordering of diverse cell components. Cellular functions are generally dictated by the biophysical phenotype of cells, which itself is also tightly regulated at the molecular level. Based on previous evidence suggesting that the relative nuclear size contributes to variations of T-cell stiffness, here we examined whether age-associated changes in T-cell migration are dictated by biophysical parameters, in part through nuclear cytoskeleton organization and cell deformability. In this study, we first performed longitudinal analyses of a repertoire of 111 functional, biophysical and biomolecular features of the nucleus and cytoskeleton of mice CD4+ and CD8+ T cells, in both naive and memory state. Focusing on the pairwise correlations, we found that age-related changes in nuclear architecture and internal ordering were correlated with T-cell stiffening and declined interstitial migration. A similarity analysis confirmed that cell-to-cell variation was a direct result of the aging process and we applied regression models to identify biomarkers that can accurately estimate individuals' age. Finally, we propose a biophysical model for a comprehensive understanding of the results: aging involves an evolution of the relative nuclear size, in part through DNA-hypomethylation and nuclear lamin B1, which implies an increased cell stiffness, thus inducing a decline in cell migration.

Mitochondrial DNA insertions into nuclear DNA affecting chromosome segregation: Insights for a novel mechanism of immunosenescence in mice.

Mitochondrial DNA sequences were found inserted in the nuclear genome of mouse peritoneal T lymphocytes that increased progressively with aging. These insertions were preferentially located at the pericentromeric heterochromatin. In the same individuals, binucleated T-cells with micronuclei showed a significantly increased frequency associated with age. Most of them were positive for centromere sequences, reflecting the loss of chromatids or whole chromosomes. The proliferative capacity of T lymphocytes decreased with age as well as the glutathione reductase activity, whereas the oxidized glutathione and malondialdehyde concentrations exhibited a significant increase. These results may point to a common process that provides insights for a new approach to understanding immunosenescence. We propose a novel mechanism in which mitochondrial fragments, originated by the increased oxidative stress status during aging, accumulate inside the nuclear genome of T lymphocytes in a time-dependent way. The primary entrance of mitochondrial fragments at the pericentromeric regions may compromise chromosome segregation, causing genetic loss that leads to micronuclei formation, rendering aneuploid cells with reduced proliferation capacity, one of the hallmark of immunosenescence. Future experiments deciphering the mechanistic basis of this phenomenon are needed.

Social environment improves the cytokine profile and lymphoproliferative response in chronologically old and prematurely aging mice.

Among the age-associated changes in the immune system, the most evident is the decrease in proliferative responses of lymphocytes to mitogenic stimuli, which is accompanied by the loss of cytokine network homeostasis. Chronic low-grade inflammatory stress, termed as sterile inflammation, is also observed during aging. In chronologically and prematurely aging mice, cohabitation with adult animals for two months favored improvements in several immune functions. This study aimed to determine whether cohabitation could restore several cytokine networks, improve lymphoproliferative responses to mitogens, and diminish sterile inflammation. Chronologically old mice (76 ± 4 weeks) and prematurely aging mice (33 ± 4 weeks) (PAM and TH-HZ) were cohabited with adults (without premature aging) for two months. Subsequently, lymphoproliferation in both basal (unstimulated) conditions and in the presence of mitogenic stimuli lipopolysaccharide A (LPS) or concanavalin A (ConA) was analyzed in cultures of peritoneal leukocytes for 48 h. Cytokine secretions (IL-1ß, TNF-α, IL-6, IL-10, and IL-17) in these cultures were also evaluated. The results showed that cohabitation restored the levels of these cytokines in old and prematurely aging mice and improved the subsequent lymphoproliferative responses. In addition, this social strategy diminished sterile inflammation and decreased inflammatory stress in unstimulated conditions. Therefore, this strategy seems to be capable of restoring the relevant immune function of lymphocytes and reducing the inflammatory stress, which are the improvements required for an adequate immune response.

Daily ingestion of Akkermansia mucciniphila for one month promotes healthy aging and increases lifespan in old female mice.

The ingestion of certain probiotics has been suggested as a promising nutritional strategy to improve aging. The objective of this work was to evaluate the effects of the daily intake, for a month, of a new probiotic Akkermansia muciniphila (AKK) (2 × 108 cfu/100µL PBS) on behavior, as well as function and redox state of immune cells of old female ICR-CD1 mice (OA group). For this, several behavioral tests were performed, and function and oxidative-inflammatory stress parameters of peritoneal leukocytes were analyzed in OA group, in a group of the same age that did not take AKK (old control, OC group) and in another adult control (AC) group. The results showed, in OA group, a significant improvement of several behavioral responses (coordination, balance, neuromuscular vigor, exploratory ability and anxiety like-behaviors), as well as in immune functions (chemotaxis, phagocytosis, NK activity and lymphoproliferation) and in oxidative stress parameters (glutathione peroxidase and reductase activities, oxidized glutathione and lipid oxidation concentrations) of the peritoneal leukocytes in comparison to those observed in OC group. In addition, peritoneal immune cells from the OA group released lower basal concentrations of pro-inflammatory cytokines (IL-2, IL-6 and TNF-α) compared to those from the OC group. The values of parameters in OA were similar to those in AC group. These improvements in the old mice receiving the probiotic were reflected in an increase in their lifespan. In conclusion, our data indicate that AKK supplementation for a short period could be a good nutritional strategy to promote healthy longevity.

A short social interaction between adult and old mice improves the homeostatic systems and increases healthy longevity.

The aging process can be influenced by environmental factors, such as the social environment. The continuous cohabitation of the chronologically old mice with adult animals improves them at the behavioral level, immune function, oxidative stress and longevity, but causes a deterioration of these parameters in adults. Therefore, the objective of the study was to study whether the coexistence for only 15 min a day of old mice with adult mice, can produce that improvement and greater longevity in old animals without causing deterioration in adults. For that, old and adult CD1 female mice, after two months of that social interaction, were submitted to a behavioral battery and then peritoneal leukocytes were collected to assess several immune functions, oxidative and inflammatory stress parameters as well as catecholamine concentrations. When the adult mice reached old age, the same parameters were again analyzed. The life span of each animal was also recorded. Plasmatic concentration of oxytocin was also studied. The results showed that old mice presented better behavioral capacity, immunity and oxi-inflammatory state after this social interaction with adult animals, and consequently an extended life span. Adult mice, in general, did not show any changes after social interaction with old animals, but when they achieved old age, improvements of some parameters and of longevity were observed in comparison with animals that never had a that social interaction. In conclusion, a short social interaction between old and adult individuals can be an excellent strategy for improving in both the health state and longevity.

The Postnatal Leptin Surge Supports Immune Cell Function in Rats.

BACKGROUND: Leptin plays an important role in the regulation of the immune response. There is a physiological surge of leptin in rodents during the neonatal period, which has mainly been studied in the context of brain development. However, little is known about the effects of this neonatal leptin surge on immunity. Therefore, we investigated whether blocking this leptin surge could affect several immune functions. METHODS: Male and female rats were injected subcutaneously with 5 mg/Kg/day of rat pegylated super leptin antagonist during the neonatal period (PND5-9). On the peripubertal period, relevant functions as well as cytokine release by spleen leukocytes were studied in these animals. RESULTS: The results showed that the animals significantly display an impaired anti-tumor NK activity and chemotactic and proliferation capacity of lymphocytes in response to mitogens. In addition, several cytokine concentrations, released under mitogen-stimulated conditions, were also altered. CONCLUSION: In conclusion, the neonatal leptin surge seems to be involved in the establishment of an adequate immune response and cytokine profile, which are crucial for the maintenance of a healthy life.

The Role of Immune Cells in Oxi-Inflamm-Aging.

Aging is the result of the deterioration of the homeostatic systems (nervous, endocrine, and immune systems), which preserve the organism's health. We propose that the age-related impairment of these systems is due to the establishment of a chronic oxidative stress situation that leads to low-grade chronic inflammation throughout the immune system's activity. It is known that the immune system weakens with age, which increases morbidity and mortality. In this context, we describe how the function of immune cells can be used as an indicator of the rate of aging of an individual. In addition to this passive role as a marker, we describe how the immune system can work as a driver of aging by amplifying the oxidative-inflammatory stress associated with aging (oxi-inflamm-aging) and inducing senescence in far tissue cells. Further supporting our theory, we discuss how certain lifestyle conditions (such as social environment, nutrition, or exercise) can have an impact on longevity by affecting the oxidative and inflammatory state of immune cells, regulating immunosenescence and its contribution to oxi-inflamm-aging.