MiRNAs that target amyloid precursor protein processing machinery in extracellular vesicles and particles derived from oral squamous cells carcinoma.

BACKGROUND: Considering that microRNAs (miRNAs), extracellular vesicles and particles (EVPs) and the amyloid precursor protein (APP) processing have been shown to be altered in oral squamous cells carcinoma (OSCC), it is possible that miRNAs that target APP processing pathways in EVPs are impacted in tumor cells. Our aim was to evaluate miRNAs that target APP itself or disintegrin and metalloproteinase domain 10 (ADAM10), which generate a trophic compound, sAPPα, in EVPs derived from OSCC cell lines, an aggressive and non-invasive, compared to normal keratinocytes. METHODS: We used two OSCC cell lines, an aggressive human oral squamous cell carcinoma cell line (SCC09) and a less aggressive cell line (CAL27) compared with a keratinocyte lineage (HaCaT). Cells were maintained in cell media, from which we isolated EVPs. EVPs were evaluated regarding their size and concentration using Nanotracking Analysis. We measured the levels of miRNAs which had as potential downstream target APP or ADAM10, specifically miR-20a-5p, miR-103a-3p, miR-424-5p, miR-92b-3p, miR-31-5p, and miR-93-5. RESULTS: There were no differences on size distributions and concentration of isolated EVPs. OSCC cell lines-derived EVPs miR-20a-5p, miR-92b-3p, and miR-93-5p were upregulated in comparison to HaCaT-derived EVPs; while miR-31-5p was reduced in EVPs obtained from CAL27 cells. CONCLUSION: Our results indicate changes in miRNAs that target APP machinery processing in EVPs derived from OSCC cell lines of different aggressiveness, which may be involved with abnormal miRNA expression in OSCC tissue and/or releasing tumor suppressor miRNA.

Potential involvement of circulating extracellular vesicles and particles on exercise effects in malignancies.

Physical activity and exercise have been widely related to prevention, treatment, and control for several non-communicable diseases. In this context, there are innumerous pre-clinical and clinical evidence indicating the potential role of exercise, beyond cancer prevention and survival, improved quality of life, including on psychological components, bone health and cachexia, from cancer survivors is described as well. This mini-review raises the potential role of circulating extracellular and particles vesicles (EVPs) cargo, as exerkines, conducting several positive effects on adjacent and/or distant tissues such as tumor, immune, bone and muscle cells. We highlighted new perspectives about microRNAs into EVPs changes induced by exercise and its benefits on malignancies, since microRNAs can be implicated with intricated physiopathological processes. Potential microRNAs into EVPs were pointed out here as players spreading beneficial effects of exercise, such as miR-150-5p, miR-124, miR-486, and miRNA-320a, which have previous findings on involvement with clinical outcomes and as well as tumor microenvironment, regulating intercellular communication and tumor growth. For example, high-intensity interval aerobic exercise program seems to increase miR-150 contents in circulating EVPs obtained from women with normal weight or overweight. In accordance circulating EVPs miR-150-5p content is correlated with prognosis colorectal cancer, and ectopic expression of miR-150 may reduce cell proliferation, invasion and metastasis. Beyond the involvement of bioactive miRNAs into circulating EVPs and their pathways related to clinical and preclinical findings, this mini review intends to support further studies on EVPs cargo and exercise effects in oncology.

Aerobic exercise, alone or combined with an anti-inflammatory drug, reduces the severity of epileptic seizures and levels of central pro-inflammatory cytokines in an animal model of epileptic seizures.

Epilepsy is a chronic neurological disorder and there is increasing evidence about the role of inflammation in epileptogenesis. These findings have spurred the search for new immunomodulatory approaches that can improve prognosis. Using an animal model of chemically-induced epileptic seizures, we tested exercise alone as non-pharmacological therapy, and exercise combined with an anti-inflammatory drug. Five groups were used: sedentary, diazepam, aerobic exercise alone, aerobic exercise combined with an anti-inflammatory drug, and naive control. Our goal was to compare the severity of the epileptic seizures between groups as well as seizure latency in a pentylenetetrazole-induced paradigm. Cytokine levels (IL-1ß, TNF-α, and IL-10) were measured. Both exercise groups showed a reduction in seizure severity and lower levels of pro-inflammatory cytokines in the cortex, while the levels of cytokines in the hippocampus remained unaffected.

Circulating Extracellular Vesicles and Particles Derived From Adipocytes: The Potential Role in Spreading MicroRNAs Associated With Cellular Senescence.

Aging is associated with adipose tissue dysfunction and is recognized as a risk factor for shortened life span. Considering that in vitro findings have shown the involvement of microRNA in extracellular vesicles and particles (EVPs) on senescence, we hypothesized that circulating EVPs derived from adipocytes can be involved in the aging process via their microRNA cargo. We aimed to determine the microRNA profiles of circulating EVPs derived from adipocytes (FABP4+) from aged and young adult animals and to perform in silico prediction of their downstream signaling effects. Plasma was obtained from Wistar rats (3 and 21 months old), and adipocyte-derived EVPs were isolated using the commercially available kit. Fatty acid-binding protein 4 (FABP4) was used for adipocyte-derived EVPs isolation; microRNA isolation and microarray expression analysis were performed. The analysis revealed 728 miRNAs, 32 were differentially between groups (p < 0.05; fold change ≥ |1.1|), of which 15 miRNAs were upregulated and 17 were downregulated in circulating EVPs from aged animals compared to young adults. A conservative filter was applied, and 18 microRNAs had experimentally validated and highly conserved predicted mRNA targets, with a total of 2,228 mRNAs. Canonical pathways, disease and functions, and upstream regulator analyses were performed using IPA-QIAGEN, allowing a global and interconnected evaluation. IPA categories impacted negatively were cell cycle, cellular development, cellular growth and proliferation, and tissue development, while those impacted positively were "digestive system cancer" and "endocrine gland tumor." Interestingly, the upregulated miR-15-5p targets several cyclins, such as CCND1 and CCND2, and miR-24-3p seems to target CDK4 (cyclin-dependent kinase 4); then potentially inhibiting their expression, both miRNAs can induce a negative regulation of cell cycle progression. In contrast, silencing of negative cell cycle checkpoint regulators, such as p21 and p16, can be predicted, which can induce impairment in response to genotoxic stressors. In addition, predicted targets, such as SMAD family members, seem to be involved in the positive control of digestive and endocrine tumors. Taken together, this exploratory study indicates that miRNA signature in circulating adipocyte-derived EVPs may be involved with the double-edged sword of cellular senescence, including irreversible proliferation arrest and tissue-dependent cancer, and seems to be suitable for further validation and confirmatory studies.

Circulating extracellular vesicles delivering beneficial cargo as key players in exercise effects.

Exercise has been recognized as an effective preventive and therapeutic approach for numerous diseases. This review addresses the potential role of circulating extracellular vesicles (EV) cargo that is modulated by physical activity. EV transport and deliver beneficial molecules to adjacent and distant tissues as a whole-body phenomenon, resulting in a healthier global status. Several candidate EV molecules, especially miRNAs, are summarized here as mediators of the beneficial effects of exercise, using different modalities, frequencies, volumes, and intensities. The following are among the candidate miRNAs: miR-21, miR-146, miR-486, miR-148a-3p, miR-223-3p, miR-142-3p, and miR-191a-5p. We highlight the relationship between EV cargo modifications, their targets and pathway interactions, in clinical outcomes, for example, on cardiovascular or immune diseases. This review brings an innovative perspective providing evidence for an intricate biological basis of the relationship between EV cargo and exercise-induced benefits on several diseases. Moreover, specific changes on circulating EV content might potentially be used as biomarkers of exercise efficacy.

The amyloid precursor protein (APP) processing as a biological link between Alzheimer's disease and cancer.

Aging is a risk factor for several illnesses, such as Alzheimer's Disease and various cancers. However, an inverse correlation between malignancies and Alzheimer's Disease has been suggested. This review addressed the potential role of non-amyloidogenic and amyloidogenic pathways of amyloid precursor protein processing as a relevant biochemical mechanism to clarify this association. Amyloidogenic and non-amyloidogenic pathways have been related to Alzheimer's Disease and certain malignancies, respectively. Several known molecules involved in APP processing, including its regulation and final products, were summarized. Among them some candidate mechanisms emerged, such as extracellular-regulated kinase (Erk) and protein kinase C (PKC). Therefore, the imbalance of APP processing may be involved with the negative correlation between cancer and Alzheimer Disease.

Circulating extracellular vesicles in the aging process: impact of aerobic exercise.

Our aim was to investigate transitory and delayed exercise effects on serum extracellular vesicles (EVs) in aging process. Male Wistar rats of 3-, 21-, and 26-month old were allocated into exercised and sedentary groups. The exercise protocol consisted in a daily moderate treadmill exercise (20 min daily during 2 weeks). Trunk blood was collected 1 and 18 h after the last exercise session, and circulating EVs were obtained. CD63 levels and acetylcholinesterase (AChE) activity were used as markers of exosome, a subtype of EVs. In addition, the quantification of amyloid-ß (Aß) levels and the oxidative status parameters, specifically reactive species content, superoxide dismutase (SOD) activity, and SOD1 content were evaluated. Aged rats showed reduced CD63 levels and increased AChE activity in circulating exosomes compared to young ones. Moreover, higher reactive species levels were found in circulating EVs of aged rats. Delayed exercise effects were observed on peripheral EVs, since CD63, reactive species content, and AChE activity were altered 18 h after the last exercise session. Our results suggest that the healthy aging process can modify circulating EVs profile, and exercise-induced beneficial effects may be related to its modulation on EVs.

Effect of tannery effluent on oxidative status of brain structures and liver of rodents.

Oxidative stress has been considered as a central mechanism of toxicity induced by xenobiotics. Previously, it was demonstrated that mice exposed to tannery effluent showed an anxiety-like behavior, without any comparable behavioral effects in rats. The aim of the present study was to investigate the impact of tannery wastewater on oxidative status in in vitro and in vivo assays with two mammal species, mice and rats. Specifically, homogenates of two brain areas and the liver were incubated with tannery wastewater; reactive species and lipid peroxidation levels and antioxidant enzyme activities were detected. In addition, the effects of in vivo exposure of mice to tannery effluents on and lipid peroxidation levels and the total reactive antioxidant capacity in brain areas and liver. Brain areas, the hippocampus and frontal cortex, and the liver of mice exposed to tannery wastewater showed oxidative stress. Our data suggest that divergent species-dependent hepatic enzymes adaptations, such as glutathione peroxidase and glutathione S-transferase activities, induced by tannery effluent could explain previous behavioral findings.

Aging process alters hippocampal and cortical secretase activities of Wistar rats.

A growing body of evidence has demonstrated amyloid plaques in aged brain; however, little attention has been given to amyloid precursor protein (APP) processing machinery during the healthy aging process. The amyloidogenic and non-amyloidogenic pathways, represented respectively by ß- and α-secretases (BACE and TACE), are responsible for APP cleavage. Our working hypothesis is that the normal aging process could imbalance amyloidogenic and non-amyloidogenic pathways specifically BACE and TACE activities. Besides, although it has been showed that exercise can modulate secretase activities in Alzheimer Disease models the relationship between exercise effects and APP processing during healthy aging process is rarely studied. Our aim was to investigate the aging process and the exercise effects on cortical and hippocampal BACE and TACE activities and aversive memory performance. Young adult and aged Wistar rats were subjected to an exercise protocol (20min/day for 2 weeks) and to inhibitory avoidance task. Biochemical parameters were evaluated 1h and 18h after the last exercise session in order to verify transitory and delayed exercise effects. Aged rats exhibited impaired aversive memory and diminished cortical TACE activity. Moreover, an imbalance between TACE and BACE activities in favor of BACE activity was observed in aged brain. Moderate treadmill exercise was unable to alter secretase activities in any brain areas or time points evaluated. Our results suggest that aging-related aversive memory decline is partly linked to decreased cortical TACE activity. Additionally, an imbalance between secretase activities can be related to the higher vulnerability to neurodegenerative diseases induced by aging.

Inflammatory response and oxidative stress in developing rat brain and its consequences on motor behavior following maternal administration of LPS and perinatal anoxia.

Cerebral palsy (CP) is a disorder of locomotion, posture and movement that can be caused by prenatal, perinatal or postnatal insults during brain development. An increased incidence of CP has been correlated to perinatal asphyxia and maternal infections during gestation. The effects of maternal exposure to low doses of bacterial endotoxin (lipopolysaccharide, LPS) associated or not with perinatal anoxia (PA) in oxidative and inflammatory parameters were examined in cerebral cortices of newborns pups. Concentrations of TNF-α, IL-1, IL-4, SOD, CAT and DCF were measured by the ELISA method. Other newborn rats were assessed for neonatal developmental milestones from day 1 to 21. Motor behavior was also tested at P29 using open-field and Rotarod. PA alone only increased IL-1 expression in cerebral cortex with no changes in oxidative measures. PA also induced a slight impact on development and motor performance. LPS alone was not able to delay motor development but resulted in changes in motor activity and coordination with increased levels of IL-1 and TNF-α expression associated with a high production of free radicals and elevated SOD activity. When LPS and PA were combined, changes on inflammatory and oxidative stress parameters were greater. In addition, greater motor development and coordination impairments were observed. Prenatal exposure of pups to LPS appeared to sensitize the developing brain to effects of a subsequent anoxia insult resulting in an increased expression of pro-inflammatory cytokines and increased free radical levels in the cerebral cortex. These outcomes suggest that oxidative and inflammatory parameters in the cerebral cortex are implicated in motor deficits following maternal infection and perinatal anoxia by acting in a synergistic manner during a critical period of development of the nervous system.

Treadmill exercise induces age-related changes in aversive memory, neuroinflammatory and epigenetic processes in the rat hippocampus.

It has been described that exercise can modulate both inflammatory response and epigenetic modifications, although the effect of exercise on these parameters during the normal brain aging process yet remains poorly understood. Here, we investigated the effect of aging and treadmill exercise on inflammatory and epigenetic parameters specifically pro and anti-inflammatory cytokines levels, activation of NF-kB and histone H4 acetylation levels in hippocampus from Wistar rats. Additionally, we evaluated aversive memory through inhibitory avoidance task. Rats of 3 and 20 months of age were assigned to non-exercised (sedentary) and exercised (running daily for 20 min for 2 weeks) groups. The effect of daily forced exercise in the treadmill was assessed. The levels of inflammatory and epigenetic parameters were determined 1h, 18 h, 3 days or 7 days after the last training session of exercise. It was observed an age-related decline on aversive memory, as well as aged rats showed increased hippocampal levels of inflammatory markers, such as TNFα, IL1-ß and NF-kB and decreased IL-4 levels, an anti-inflammatory cytokine. Moreover, lower levels of global histone H4 acetylation were also observed in hippocampi from aged rats. Interestingly, there was a significant correlation between the biochemical markers and the inhibitory avoidance test performance. The forced exercise protocol ameliorated aging-related memory decline, decreased pro-inflammatory markers and increased histone H4 acetylation levels in hippocampi 20-months-old rats, while increased acutely IL-4 levels in hippocampi from young adult rats. Together, these results suggest that an imbalance of inflammatory markers might be involved to the aging-related aversive memory impairment. Additionally, our exercise protocol may reverse aging-related memory decline through improving cytokine profile.

[Evaluation of the impact of exposure to pesticides on the health of the rural population: Vale do Taquari, State of Rio Grande do Sul (Brazil)]. / Avaliação do impacto da exposição a agrotóxicos sobre a saúde de população rural: Vale do Taquari (RS, Brasil).

The scope of this cross-sectional study was to assess a possible link between contact with pesticides and the prevalence of chronic disease in the rural population in the south of Brazil. Three municipalities were randomly chosen. The sample selected was recruited over three months (2005) and was composed of 298 subjects of both sexes, between 18 and 65 years of age, who frequented public or private pharmacies for the purchase of medication. Data obtained by means of structured interviews revealed that 68.4% of those interviewed worked in rural activities, and 74.8% of these were members of families working in agriculture with contact with pesticides. The average age was 51+16.5 years of age for the interviewees with contact with pesticides and 50+17.9 years for those without contact. The sample was composed of 36.2% of men, and 57.7% of the individuals had more than four years of education. Direct or indirect contact with pesticides was associated with the report of several diseases, with neurological and oral diseases being the most prevalent. There was a link with painful conditions and individuals with contact with pesticides reported twice as many diseases. The data corroborate the literature and draw attention to the need for promoting measures to protect the health of the rural population.

Avaliação do impacto da exposição a agrotóxicos sobre a saúde de população rural: Vale do Taquari (RS, Brasil) / Evaluation of the impact of exposure to pesticides on the health of the rural population: Vale do Taquari, State of Rio Grande do Sul (Brazil)

Este estudo transversal teve o objetivo de avaliar possível associação entre contato com agrotóxicos e prevalência de doenças crônicas em população rural do Sul do Brasil. Três municípios foram aleatoriamente escolhidos. A amostra selecionada por conveniência foi recrutada durante três meses (2005) e se compôs de 298 sujeitos de ambos os sexos, com idade entre 18 e 65 anos e que procuravam farmácia pública ou privada para a compra de medicamentos. Os dados obtidos, mediante entrevistas estruturadas, demonstraram que 68,4 por cento dos entrevistados exerciam atividade rural, dos quais 74,8 por cento eram membros de famílias de agricultores e tinham contato com agrotóxicos. A média de idade foi 51±16,5 anos para os entrevistados com contato com agrotóxicos e 50±17,9 anos para os sem contato. A amostra foi constituída de 36,2 por cento por homens e 57,7 por cento dos indivíduos que apresentavam mais de quatro anos de estudo. O contato direto ou indireto com agrotóxicos associou-se ao relato de várias doenças, sendo as neurológicas e as orais as mais prevalentes. Houve associação com relato de condições dolorosas, de modo que indivíduos com contato com agrotóxicos apresentaram em torno de duas vezes mais chances de as referirem. Os dados corroboram os da literatura e indicam a necessidade de promoção de medidas de proteção e prevenção da saúde da população rural.

The scope of this cross-sectional study was to assess a possible link between contact with pesticides and the prevalence of chronic disease in the rural population in the south of Brazil. Three municipalities were randomly chosen. The sample selected was recruited over three months (2005) and was composed of 298 subjects of both sexes, between 18 and 65 years of age, who frequented public or private pharmacies for the purchase of medication. Data obtained by means of structured interviews revealed that 68.4 percent of those interviewed worked in rural activities, and 74.8 percent of these were members of families working in agriculture with contact with pesticides. The average age was 51+16.5 years of age for the interviewees with contact with pesticides and 50+17.9 years for those without contact. The sample was composed of 36.2 percent of men, and 57.7 percent of the individuals had more than four years of education. Direct or indirect contact with pesticides was associated with the report of several diseases, with neurological and oral diseases being the most prevalent. There was a link with painful conditions and individuals with contact with pesticides reported twice as many diseases. The data corroborate the literature and draw attention to the need for promoting measures to protect the health of the rural population.

Different intensities of treadmill running exercise do not alter melatonin levels in rats / Diferentes intensidades de exercício de corrida em esteira não alteram os níveis de melatonina em ratos

Background: Regular and moderate exercise has been considered an interesting neuroprotective strategy. Our research group demonstrated that a protocol of moderate exercise on a treadmill reduced, while a protocol of high-intensity exercise increased in vitro ischemic cell damage in Wistar rats. The molecular mechanisms by which physical exercise exerts neuroprotective effects remain unclear. Accumulating evidence suggests that exercise may have short- and long-term effects on melatonin secretion in humans. Melatonin, the main product of the pineal gland, has been shown to have neuroprotective effects in models of brain and spinal cord injury and cerebral ischemia. A dual modulation of melatonin secretion by physical activity has also been demonstrated. This study aimed to investigate the effect of different exercise intensities, moderateand high-intensity exercise, on serum melatonin levels in rats. Methods: Thirty-five adult male Wistar rats were divided into non-exercised (sedentary) and exercised (20- or 60-min sessions) groups. The exercise protocols consisted of two weeks of daily treadmill training. Blood samples were collected approximately 16 hours after the last training session (8:00-10:00) and melatonin levels were assayed by ELISA. Results: The exercise protocols, two weeks of 20 min/day or 60 min/day of treadmill running, did not affect serum melatonin levels. Conclusion: Our data demonstrated that melatonin levels may not be directly involved in the exercise-induced, intensitydependent dual effect on in vitro ischemia.

Introdução: O exercício físico regular e moderado é considerado uma interessante estratégia neuroprotetora. Nosso grupo de pesquisa demonstrou que um protocolo de exercício moderado em esteira reduziu, enquanto um protocolo de intensidade elevada aumentou o dano isquêmico in vitro em ratos Wistar. Os mecanismos moleculares pelos quais o exercício físico exerce neuroproteção ainda não estão claramente elucidados. Várias evidências sugerem que o exercício pode ter efeitos a curto e longo prazo sobre a secreção de melatonina em humanos. A melatonina, o principal produto da glândula pineal, tem demonstrado exercer uma atividade neuroprotetora em modelos de trauma cerebral e medular e isquemia cerebral. Foi demonstrada também uma modulação dual da atividade física sobre a secreção de melatonina. O objetivo deste estudo foi estudar o efeito de diferentes intensidades de exercício, moderado e elevado, sobre os níveis séricos de melatonina de ratos. Métodos: Trinta e cinco ratos Wistar adultos machos foram divididos em não-exercitados (sedentários) ou exercitados (sessões de 20 ou 60 min). Os protocolos de exercício consistiam em duas semanas diárias de treinamento em esteira ergométrica. Os soros foram coletados aproximadamente 16 horas após a última sessão de treino (8:00-10:00) e os níveis de melatonina foram avaliados através da técnica de ELISA. Resultados: Os protocolos de exercício físico, duas semanas de 20 min/dia ou 60 min/dia de corrida em esteira, não alteraram os níveis séricos de melatonina. Conclusão: Nossos dados demonstram que os níveis de melatonina parecem não estar diretamente envolvidos com o efeito dual do exercício físico dependente da intensidade na isquemia in vitro.

A neuroprotective exercise protocol reduces the adenine nucleotide hydrolysis in hippocampal synaptosomes and serum of rats.

Regular and moderate exercise has been considered as an interesting neuroprotective strategy. However, the molecular mechanisms by which physical exercise alters brain function are unclear. Purinergic signaling seems to modulate the pathophysiology of ischemic neuronal damage, since it has been described a neuroprotective activity of adenosine and a dual role of ATP. In the present study, we investigated the effect of daily moderate intensity exercise on ectonucleotidase activities in synaptosomes from hippocampus and the soluble nucleotidases from blood serum of rats. Adult male Wistar rats were assigned to non-exercised (sedentary) group and exercised during 20-min sessions on different programs. The effects of physical activity on hydrolysis of ATP, ADP and AMP were assayed in the synaptosomal fraction obtained from the hippocampus and serum approximately 16 h after the last training session. Our data demonstrated that a neuroprotective exercise protocol, daily 20 min of training in treadmill during 2 weeks, diminished significantly the ADP hydrolysis and there is a trend to reduce the ATP hydrolysis in both hippocampal synaptosomes and blood serum of rats. We suggest that the neuroprotective exercise protocol may modulate nucleotidase activities.