Extracellular vesicles: Methods for purification and characterization.

Extracellular vesicles (EVs) are membrane-bound particles released by cells that play a significant role in intercellular communication. They can be obtained from a variety of sources, including conditioned culture medium, blood and urine. In this chapter we detail the methods for EV isolation and characterization. Isolating and characterizing EVs is essential for understanding their functions in physiological and pathological processes. Advances in isolation and characterization techniques provide opportunities for deeper research into EV biology and its potential applications in diagnostics and therapeutics.

Extracellular vesicles.

Cells, pathogens, and other systems release extracellular vesicles (EVs). The particles promote intercellular communication and contain proteins, lipids, RNA and DNA. Initially considered to be cellular waste in the twentieth century, EVs were becoming recognized for their function in biological communication and control. EVs are divided into many subtypes: exosomes, microvesicles, and apoptotic bodies. Exosomes form in the late endosome/multivesicular body and are released when the compartments fuse with the plasma membrane. Microvesicles are generated by direct budding of the plasma membrane, whereas apoptotic bodies are formed after cellular apoptosis. The new guideline for EVs that describes alternate nomenclature for EVs. The particles modulate the immune response by affecting both innate and adaptive immunity, and their specific the structure allows them to be used as biomarkers to diagnose a variety of diseases. EVs have a wide range of applications, for example, delivery systems for medications and genetic therapies because of their ability to convey specific cellular material. In anti-tumor therapy, EVs deliver therapeutic chemicals to tumor cells. The EVs promote transplant compatibility and reduce organ rejection. Host-parasite interactions, therapeutic and diagnostic for cancer, cardiovascular disease, cardiac tissue regeneration, and the treatment of neurological diseases such as Alzheimer's and Parkinson's. The study of EVs keeps on expanding, revealing new functions and beneficial options. EVs have the potential to change drug delivery, diagnostics, and specific therapeutics, creating a new frontier in biomedical.

Linking human and ecological components to understand human-wildlife conflicts across landscapes and species.

Human-wildlife conflicts (HWC) are complex conservation challenges that impair both wildlife populations and human livelihood. Research on HWC, however, has traditionally approached ecological and human components separately, hampering a broader understanding of connections between ecological drivers and human dimensions of conflicts. We developed a model that integrates ecological and human components of HWC to investigate how the amount of remaining native forest (forest cover, a key ecological variable known to influence species occurrence and abundance) affects human experiences with wildlife (contact with species and attacks on livestock) and how such experiences influence tolerance via beliefs, emotions, and attitudes. We tested the model with piecewise structural equation modeling and data on human interactions with 3 mammals with different rarity and body size: opossum (Didelphis aurita), crab-eating fox (Cerdocyon thous), and puma (Puma concolor). Data were obtained by interviewing 114 landowners across 13 Atlantic Forest landscapes (10-50% forest cover). Forest cover was associated with high chance of attacks on livestock, and thus with low tolerance, only in the case of the puma. Effects of distinct experiences with wildlife on beliefs and emotions varied across species. Beliefs and emotions toward wildlife influenced tolerance toward all species, but negative emotions affected tolerance toward only with the puma. Conflicts with large carnivores, such as pumas, can then be understood as disservices provided by forests, indicating the relevance of framing HWC more broadly to consider trade-offs with ecosystems services. For some species, positive experiences with wildlife may counteract the negative effects of attacks on livestock in shaping tolerance. Models such as ours-that link ecological and human dimensions-can help identify more effective leverage points to improve HWC mitigation.

Vinculación de componentes humanos y ecológicos para entender los conflictos humano-fauna a nivel paisaje y especies Resumen Los conflictos humano-fauna (CHF) son retos complejos para la conservación pues perjudican tanto a las poblaciones de fauna como al sustento humano. Sin embargo, las investigaciones sobre los CHF por tradición han manejado a los componentes humanos y ecológicos de manera separada, dificultando un entendimiento más amplio de las conexiones entre los conductores ecológicos y las dimensiones humanas de estos conflictos. Desarrollamos un modelo que integra a los componentes ecológicos y humanos de los CHF para investigar cómo la cantidad de bosque nativo remanente (la cobertura de bosque, una variable ecológica importante que se sabe influye sobre la presencia y abundancia de las especies) afecta a las experiencias que las personas tienen con la fauna (contacto con especies y ataques al ganado) y cómo dichas experiencias influyen en la tolerancia por medio de creencias, emociones y actitudes. Probamos el modelo con un modelado de ecuación estructural por partes y datos sobre las interacciones humanas con tres mamíferos de diferente tamaño y rareza: la zarigüeya (Didelphis aurita), el zorro cangrejero (Cerdocyon thous) y el puma (Puma concolor). Los datos se obtuvieron por medio de entrevistas realizadas a 114 terratenientes distribuidos en 13 paisajes de bosque atlántico (10-50% de cobertura de bosque). La cobertura de bosque estuvo asociada con una alta probabilidad de ataques al ganado, y por lo tanto con una baja tolerancia, sólo para el caso del puma. Los efectos de las distintas experiencias con la fauna sobre las creencias y las emociones variaron con cada especie. Las creencias y las emociones hacia la fauna influyeron en la tolerancia hacia todas las especies, aunque las emociones negativas afectaron la tolerancia solamente hacia el puma. Los conflictos con los grandes carnívoros, como el puma, pueden entenderse como servicios negativos proporcionados por los bosques, lo que indica la relevancia de enmarcar a los CHF de manera más amplia para considerar las compensaciones con los servicios ambientales. Para algunas especies, las experiencias positivas con la fauna pueden contrarrestar los efectos negativos de los ataques al ganado en cuanto a la formación de la tolerancia. Los modelos como el nuestro - que conectan las dimensiones ecológicas y humanas - pueden ayudar a identificar puntos de ventaja más efectivos para mejorar la mitigación de los CHF.

The influence of fluoxetine on orthodontic tooth movement in rats.

This study aimed to evaluate the effects of chronic use of fluoxetine on the amount of orthodontic tooth movement and tissue changes in rats. A total of 192 Wistar rats were divided into 4 groups: S, 0.9% saline solution; F, 20 mg/kg of fluoxetine; SM, 0.9% saline solution with orthodontic movement; and FM, 20 mg/kg of fluoxetine with orthodontic movement. After 30 days of daily saline or fluoxetine administration, an orthodontic device (25cN) was used to mesially displace the first molar in animals of the groups SM and FM. The animals were euthanized 2, 7, 14, and 28 days after placement of the orthodontic appliances and animals of groups S and F were euthanized at the same time. The assessment of tooth movement was made in gypsum castings, the collagen neoformation was assessed by polarization microscopy, the number of osteoclasts and root resorption were evaluated using tartrate-resistant acid phosphatase, and presence of hyalinized areas was assessed by hematoxylin-eosin staining. Fluoxetine did not affect the amount of tooth displacement, percentage of collagen, number of osteoclasts, and presence of hyalinized areas (P>0.05). There was a higher frequency of root resorption areas in the FM group than in the SM group only on the second day (P<0.05). The findings of this study show that chronic use of 20 mg/kg fluoxetine does not affect the amount of tooth movement, collagen neoformation, number of osteoclasts, or hyalinized areas and does not affect root resorption until the last day of orthodontic movement.

A mango (Mangifera indica L.) juice by-product reduces gastrointestinal and upper respiratory tract infection symptoms in children.

The study aimed to evaluate the effect of a mango juice by-product (JBP) on upper-respiratory and gastrointestinal tract infection symptoms in children (6-8 y) in a randomized, double-blind, parallel, case-control study. For two months, children drank either flavored water (control group) or a mango JBP-based beverage (0.04 g·ml-1; treatment group); such beverage provided 1.1 g, 278.6 mg and 7.8 mg of dietary fiber, extractable polyphenols (mono-to-hepta galloyl hexosides, mangiferin), and hydrolysable polyphenols (ellagic/gallic acid) per portion, respectively. Mango JBP reduced the incidence of gastrointestinal (flatulencies and abdominal inflammation; p ≤ 0.007) and upper-tract respiratory (crystalline mucus, itchy throat, runny nose, itchy nose, and sneezing; p ≤ 0.038) and such benefits were associated to increased serum levels of PAI-I, MIP-1a, and MIP-1b (p ≤ 0.04) and decreased levels of IgG, MIF, and osteopontin (p ≤ 0.01). We concluded that JBP-based beverage has immunomodulatory properties, useful to prevent or even treat common infectious diseases in school-age children.

Bacillus coagulans GBI-30, 6068 decreases upper respiratory and gastrointestinal tract symptoms in healthy Mexican scholar-aged children by modulating immune-related proteins.

This randomized, double-blind, parallel and placebo-controlled study aimed to evaluate the effect of Bacillus coagulans GBI-30, 6086® probiotic (GanedenBC30®) against upper respiratory tract infections (URTI) and gastrointestinal tract infections (GITI) in eighty healthy school-aged children (6-8 years old). The participants received daily a sachet containing either GanedenBC30 (1 × 109 colony-forming units) or placebo (maltodextrin) for three months. GanedenBC30 significantly decreased the incidence of URTI symptoms including nasal congestion, bloody nasal mucus, itchy nose, and hoarseness. The duration of the URTI-associated symptoms of hoarseness, headache, red eyes, and fatigue was also decreased. GanedenBC30 supplementation also significantly reduced the incidence rate of flatulence. These beneficial effects were associated with the modulation of serum TNFα, CD163, G-CSF, ICAM-1, IL-6, IL-8, MCP-2, RAGE, uPAR, and PF4. Therefore, probiotic B. coagulans GBI-30, 6086 modulated immune-related proteins in healthy children, decreasing several URTI and GITI symptoms, thus, this functional ingredient may contribute to a healthier lifestyle.

The influence of fluoxetine on orthodontic tooth movement in rats

Abstract This study aimed to evaluate the effects of chronic use of fluoxetine on the amount of orthodontic tooth movement and tissue changes in rats. A total of 192 Wistar rats were divided into 4 groups: S, 0.9% saline solution; F, 20 mg/kg of fluoxetine; SM, 0.9% saline solution with orthodontic movement; and FM, 20 mg/kg of fluoxetine with orthodontic movement. After 30 days of daily saline or fluoxetine administration, an orthodontic device (25cN) was used to mesially displace the first molar in animals of the groups SM and FM. The animals were euthanized 2, 7, 14, and 28 days after placement of the orthodontic appliances and animals of groups S and F were euthanized at the same time. The assessment of tooth movement was made in gypsum castings, the collagen neoformation was assessed by polarization microscopy, the number of osteoclasts and root resorption were evaluated using tartrate-resistant acid phosphatase, and presence of hyalinized areas was assessed by hematoxylin-eosin staining. Fluoxetine did not affect the amount of tooth displacement, percentage of collagen, number of osteoclasts, and presence of hyalinized areas (P>0.05). There was a higher frequency of root resorption areas in the FM group than in the SM group only on the second day (P<0.05). The findings of this study show that chronic use of 20 mg/kg fluoxetine does not affect the amount of tooth movement, collagen neoformation, number of osteoclasts, or hyalinized areas and does not affect root resorption until the last day of orthodontic movement.

Evoked Potentials and Memory/Cognition Tests Validate Brain Atrophy as Measured by 3T MRI (NeuroQuant) in Cognitively Impaired Patients.

To our knowledge, this is the largest study evaluating relationships between 3T Magnetic Resonance Imaging (MRI) and P300 and memory/cognitive tests in the literature. The 3T MRI using NeuroQuant has an increased resolution 15 times that of 1.5T MRI. Utilizing NeuroQuant 3T MRI as a diagnostic tool in primary care, subjects (N=169; 19-90 years) displayed increased areas of anatomical atrophy: 34.62% hippocampal atrophy (N=54), 57.14% central atrophy (N=88), and 44.52% temporal atrophy (N=69). A majority of these patients exhibited overlap in measured areas of atrophy and were cognitively impaired. These results positively correlated with decreased P300 values and WMS-III (WMS-III) scores differentially across various brain loci. Delayed latency (p=0.0740) was marginally associated with temporal atrophy; reduced fractional anisotropy (FA) in frontal lobes correlated with aging, delayed P300 latency, and decreased visual and working memory (p=0.0115). Aging and delayed P300 latency correlated with lower FA. The correlation between working memory and reduced FA in frontal lobes is marginally significant (p=0.0787). In the centrum semiovale (CS), reduced FA correlated with visual memory (p=0.0622). Lower demyelination correlated with higher P300 amplitude (p=0.0002). Compared to males, females have higher demyelination (p=0.0064). Along these lines, the higher the P300 amplitude, the lower the bilateral atrophy (p=0.0165). Hippocampal atrophy correlated with increased auditory memory and gender, especially in males (p=0.0087). In considering temporal lobe atrophy correlations: delayed P300 latency and high temporal atrophy (p=0.0740); high auditory memory and low temporal atrophy (p=0.0417); and high working memory and low temporal atrophy (p=0.0166). Central atrophy correlated with aging and immediate memory (p=0.0294): the higher the immediate memory, the lower the central atrophy. Generally, the validation of brain atrophy by P300 and WMS-III could lead to cost-effective methods utilizable in primary care medicine following further confirmation.

Vasculitis como presentación de leucemia promielocítica aguda / Vasculitis as a presentation of acute promyelocytic leukemia

Acute promyelocytic leukemiais a subtype of acute my eloid leukemia characterized by the presence of the PML:RAR aonco proteindueto a specific geneticalteration, translocation (15;17), which avoids my eloid differentiation, generating the accumulation of leukemic promyelocytes with posterior alteration of hemostasis. It presents with symptoms related to pancytopenia and thrombohemorrhagic coagulopathy, and there are few report sof cases with vasculitis secondary to promyelocytic leukemia. The clinical case aimed to describe a particular form of presentation of acute promyelocytic leukemia with vasculitis.

La leucemia promielocítica aguda es un subtipo de leucemia mieloide aguda caracterizada por la presencia de la oncoproteína PML-RARa debido a una alteracióngenéticaespecífica, la translocación (15;17), que impide la diferenciación mieloide generando la acumulación de promielocitos leucémicos y una alteración compleja de la hemostasia. Se presenta con síntomas relacionados a pancitopenia y coagulopatía trombo-hemorrágica existiendo escasos reportes de casos de vasculitis asociada a leucemia. El caso clínico descrito representa una forma particular de presentación de una leucemia promielocítica aguda con vasculitis.

A intrusão isolada dos incisivos centrais superiores na má oclusão Classe II, divisão 2 – relato de caso / An individual intrusion of maxillary central incisors in Angle Class II, division 2 – case report

A má oclusão Classe II divisão 2 de Angle é caracterizada pela relação molar de Classe II associada a um posicionamento vertical ou retroinclinado dos incisivos superiores e, geralmente, associada a uma sobremordida exagerada. No presente caso clínico com tais características foi, inicialmente, intruído os incisivos centrais superiores com o arco de intrusão preconizado por Burstone. Após o tratamento corretivo, os objetivos de função, saúde e estética adequada foram plenamente alcançados...

Angle Class II division 2 malocclusion is characterized by a Class II molar relation associated with retroclined or vertical positioning of the upper incisors and usually associated with overbite. The present study reports a clinical case with the previous characteristics in which the initial step was the intrusion of maxillary central incisors using the Burstone intrusion archwire. After orthodontic treatment proper function, health and aesthetics were fully achieved...