Análise do transcriptoma de diferentes tipos de células únicas do sangue periférico de pacientes infectados com o vírus da Dengue para detalhamento da massiva geração de células secretoras de anticorpos / Evaluation of the tryptophan metabolism in the massive generation of antibody-secreting cells in Dengue patients through the transcriptome of single blood cells

A Dengue induz uma resposta exacerbada e transitória das células secretoras de anticorpos (ASCs) no sangue de pacientes cerca de sete dias após o início dos sintomas. A frequência dessas ASCs chega a representar mais de 50% de todas as células B circulantes neste período. No entanto, ainda é desconhecido se a magnitude dessa resposta tem relação com a gravidade da Dengue. Nosso grupo de pesquisa já mostrou que a cultura de células nucleares do sangue periférico (PBMCs) de indivíduos saudáveis com partículas do vírus da Dengue (DENV), por 7 dias, levava a diferenciação de células B em ASCs em magnitude similar àquelas estimuladas por mitógenos. Essas culturas apresentavam um consumo significativamente maior de triptofano (TRP), associado à maior expressão das enzimas IDO1 e IDO2, e, consequentemente, maior síntese de quinurenina (KYN) em relação ao estímulo por mitógenos. Considerando que as concentrações de TRP e KYN detectadas nos sobrenadantes dessas culturas eram diretamente proporcionais ao aumento de ASCs, decidimos investigar o papel desse metabolismo do TRP e de seus respectivos metabólitos na diferenciação das ASCs. Para isso, análises do transcriptoma público de células únicas do sangue periférico de pacientes com Dengue (estudo E-MTAB- 9467) foram realizadas para inferir a real participação do metabolismo do TRP na geração de ASCs. Com o programa R foram executadas análises de Downstream. Identificamos um aumento massivo das ASCs nas amostras dos pacientes infectados com Dengue. No entanto, os principais genes desencadeadores da ativação do metabolismo do TRP (IDO1 e IDO2) não foram expressos nas subpopulações de células B, mas sim em células dendríticas e monócitos CD14+ respectivamente. Isso sugeriria que esta via não seria ativada nos linfócitos B. Por outro lado, genes codificadores de outros participantes da via do TRP (HSD17B10, ECHS1 e SIRT3) foram detectados em células B e podem estar relacionados com a proliferação das ASCs. Além disso, a análise de enriquecimento mostrou uma aumentada expressão de genes associados com moléculas de MHC de classe II em plasmablastos e plasmócitos de pacientes com Dengue. Porém, com a expressão aumentada de ENTPD1 nessas células durante a fase sintomatológica, nossos dados sugerem também que um eventual papel de plasmablastos e plasmócitos como apresentadoras de antígenos na Dengue poderia induzir uma resposta supressora de células T

Dengue can cause an exacerbated and transient antibody-secreting cell (ASC) response in the blood of patients nearly 7 days of symptomatology. The ASC frequency reaches more than 50% of all circulating B cells during that period. However, it is still unknown whether the magnitude of this response may be directly related to the severity of Dengue. Our research group has already shown that the culture of peripheral blood mononuclear cells (PBMCs) from healthy individuals with Dengue virus (DENV) particles for 7 days led to a differentiation of B cells into ASCs to a magnitude similar to those stimulated by mitogens. These cultures showed significantly higher consumption of tryptophan (TRP), associated with higher expression of enzymes IDO1 and IDO2, and consequently, higher synthesis of quinurenine (KYN) compared to mitogen stimulation. The concentrations of TRP and KYN detected in the supernatants of these cultures were directly proportional to ASC frequency increase. Thus, we have decided to investigate the role of TRP metabolism and its respective metabolites in ASC differentiation. For this, we performed an analysis of single-cell transcriptome with peripheral blood from Dengue patients (dataset from E-MTAB-9467 study). Downstream analyses were performed with R software. Corroborating with literature, we identified a massive increase in ASC frequency of Dengue infected patients. However, the main genes triggering TRP metabolism activation (IDO1 and IDO2) were not expressed in B-cell subsets, but in dendritic cells and CD14+ monocytes, respectively. This would suggest that this pathway would not be activated in B lymphocytes. Nevertheless, genes encoding other participants in the TRP pathway (HSD17B10, ECHS1, and SIRT3) were detected in B cells and may be related to ASC proliferation. Futhermore, a Gene Ontology analysis showed an increased expression of genes associated with MHC class II molecules in plasmablasts and plasma cells of Dengue patients. As these cells also presented an increased expression of ENTPD1 during the symptomatic phase, our data suggest a potential role of plasmablasts and plasma cells as antigen-presenting cells associated with a suppressive T cell response in Dengue

Confirmation of the presence of serotonergic cells in the fetal cerebral cortex in cultures and in situ.

BACKGROUND: Early appearance of serotonin in the fetal brain and its effects on brain morphogenesis support its neurotrophic role. OBJECTIVE: To determine the presence of serotonergic cells and the expression of tryptophan-5-hydroxylase (TPH), 5-hydroxytryptamine (5-HT), serotonin transporter (SERT), 5-HT1A receptor and Pet-1 during the development of the cerebral cortex, both in situ and in tissue cultures. MATERIAL AND METHODS: A descriptive, observational study was carried out in pregnant Wistar rats. The presence of the plug was regarded as the beginning of gestation. On days 13, 16 and 17, cesarean sections were performed to obtain the fetuses, and the brains were then immediately dissected to identify the presence of serotonergic cells, TPH, 5-HT, SERT, 5-HT1A and Pet-1 in tissue cultures and in situ by immunostaining detected on a confocal microscope. RESULTS: Serotonergic cells and terminals were observed in the midbrain on day 17 of gestation, and in neopallium cocultures on days 13 and 16. TPH, 5-HT, SERT and Pet-1 immunopositive cells were also observed in the neopallium on day 12 of culture. CONCLUSIONS: The presence of serotonergic cells and other elements of the serotonergic system in the early cerebral cortex was confirmed, which may be transient and participate in cortical maturation processes during brain development.

ANTECEDENTES: La aparición temprana de serotonina en el cerebro fetal y sus efectos en la morfogénesis cerebral apoyan su papel neurotrófico. OBJETIVO: Determinar la presencia de células serotoninérgicas y la expresión de triptófano-5-hidroxilasa (TPH), 5-hidroxitriptamina (5-HT), transportador de serotonina (SERT), receptor 5-HT1A y Pet-1 durante el desarrollo de la corteza cerebral, tanto in situ como en cultivo de tejidos. MATERIAL Y MÉTODOS: Se realizó estudio observacional descriptivo en ratas Wistar preñadas. La presencia del tapón se consideró el inicio de la gestación; en los días 13, 16 y 17 se practicaron cesáreas para obtener los fetos e inmediatamente se disecaron los cerebros para identificar células serotoninérgicas, TPH, 5-HT, SERT, 5-HT1A y Pet-1 en cultivo de tejido e in situ mediante inmunomarcaje detectado en un microscopio confocal. RESULTADOS: Células y terminales serotoninérgicas fueron observadas en el mesencéfalo el día 17 de gestación y en cocultivos de neopalio los días 13 y 16. También se observaron células inmunopositivas a TPH, 5-HT, SERT y Pet-1 en el neopalio en el día 12 del cultivo. CONCLUSIONES: Se confirmó la presencia de células serotoninérgicas y otros elementos del sistema serotoninérgico en la corteza cerebral temprana, la cual puede ser transitoria y participar en los procesos de maduración cortical durante el desarrollo cerebral.

[Properties of milk in sleep induction]. / Propiedades de la leche en la inducción del sueño.

Introduction: Sleep induction and its quality are issues of growing concern because its deterioration affects a large number of people and poses a risk to their well-being and quality of life and long-term health. There are several factors involved in the problem, but nutrition is one of them and in particular milk consumption has often been linked to sleep habits, sometimes as a promoter and sometimes as an inhibitor. The purpose of this review is to examine the matter further. On reaching the brain, tryptophan is the basis for the synthesis of serotonin and melatonin, which improve the induction and quality of sleep. But there is competition between tryptophan and other long-chain neutral amino acids (LNAA) (valine, leucine, isoleucine, tyrosine and phenylalanine) to cross the blood-brain barrier and reach the brain. In this sense, milk proteins with a high tryptophan content and the highest ratio between tryptophan and LNAA are very useful in promoting sleep. Moreover, milk also provides various micronutrients that help in the transformation of tryptophan into serotonin and melatonin, as well as antioxidant components, anti-inflammatory and bioactive peptides, and recent studies indicate that it favorably modulates the composition of the intestinal microbiota. Studies show that increasing milk consumption, up to the recommended intake and within a correct diet, favors the achievement and maintenance of quality sleep.

Introducción: La inducción del sueño y su calidad son temas de preocupación creciente porque su deterioro afecta a un número elevado de personas y supone un riesgo en su bienestar y calidad de vida y en la salud a largo plazo. Hay diversos factores implicados en el problema, pero la nutrición es uno de ellos y, en concreto, el consumo de leche se ha relacionado frecuentemente con los hábitos de sueño, a veces como factor promotor y otras como inhibidor. Profundizar en el tema es el objeto de la presente revisión. El triptófano, al llegar al cerebro, es la base para la síntesis de serotonina y melatonina, que mejoran la inducción y la calidad del sueño. Pero hay una competencia entre el triptófano y otros aminoácidos neutros de cadena larga (ANCL) (valina, leucina, isoleucina, tirosina y fenilalanina) para cruzar la barrera hematoencefálica y llegar al cerebro. En este sentido, las proteínas de la leche, con elevado contenido en triptófano y la relación más elevada entre triptófano y ANCL, son muy útiles en la promoción del sueño. Por otra parte, la leche también aporta diversos micronutrientes que ayudan en la transformación del triptófano en serotonina y melatonina, así como componentes antioxidantes, antiinflamatorios y péptidos bioactivos, y estudios recientes indican que modula favorablemente la composición de la microbiota intestinal. Los estudios realizados ponen de relieve que aumentar el consumo de leche, hasta el aporte aconsejado y dentro de una alimentación correcta, favorece el conseguir y mantener un sueño de calidad.

Confirmación de la presencia de células serotoninérgicas en la corteza cerebral fetal en cultivos e in situ / Confirmation of the presence of serotonergic cells in the fetal cerebral cortex in cultures and in situ

Resumen Antecedentes: La aparición temprana de serotonina en el cerebro fetal y sus efectos en la morfogénesis cerebral apoyan su papel neurotrófico. Objetivo: Determinar la presencia de células serotoninérgicas y la expresión de triptófano-5-hidroxilasa (TPH), 5-hidroxitriptamina (5-HT), transportador de serotonina (SERT), receptor 5-HT1A y Pet-1 durante el desarrollo de la corteza cerebral, tanto in situ como en cultivo de tejidos. Material y métodos: Se realizó estudio observacional descriptivo en ratas Wistar preñadas. La presencia del tapón se consideró el inicio de la gestación; en los días 13, 16 y 17 se practicaron cesáreas para obtener los fetos e inmediatamente se disecaron los cerebros para identificar células serotoninérgicas, TPH, 5-HT, SERT, 5-HT1A y Pet-1 en cultivo de tejido e in situ mediante inmunomarcaje detectado en un microscopio confocal. Resultados: Células y terminales serotoninérgicas fueron observadas en el mesencéfalo el día 17 de gestación y en cocultivos de neopalio los días 13 y 16. También se observaron células inmunopositivas a TPH, 5-HT, SERT y Pet-1 en el neopalio en el día 12 del cultivo. Conclusiones: Se confirmó la presencia de células serotoninérgicas y otros elementos del sistema serotoninérgico en la corteza cerebral temprana, la cual puede ser transitoria y participar en los procesos de maduración cortical durante el desarrollo cerebral.

Abstract Background: Early appearance of serotonin in the fetal brain and its effects on brain morphogenesis support its neurotrophic role. Objective: To determine the presence of serotonergic cells and the expression of tryptophan-5-hydroxylase (TPH), 5-hydroxytryptamine (5-HT), serotonin transporter (SERT), 5-HT1A receptor and Pet-1 during the development of the cerebral cortex, both in situ and in tissue cultures. Material and methods: A descriptive, observational study was carried out in pregnant Wistar rats. The presence of the plug was regarded as the beginning of gestation. On days 13, 16 and 17, cesarean sections were performed to obtain the fetuses, and the brains were then immediately dissected to identify the presence of serotonergic cells, TPH, 5-HT, SERT, 5-HT1A and Pet-1 in tissue cultures and in situ by immunostaining detected on a confocal microscope. Results: Serotonergic cells and terminals were observed in the midbrain on day 17 of gestation, and in neopallium cocultures on days 13 and 16. TPH, 5-HT, SERT and Pet-1 immunopositive cells were also observed in the neopallium on day 12 of culture. Conclusions: The presence of serotonergic cells and other elements of the serotonergic system in the early cerebral cortex was confirmed, which may be transient and participate in cortical maturation processes during brain development.

Triptófano-5-hidroxilasa, transportador de serotonina y receptores serotoninérgicos en miocardiopatía dilatada.

BACKGROUND: Cardiomyocytes synthesize, utilize and reuptake serotonin, which is involved in the paracrine and autocrine modulation of heart activity and in the pathophysiology of some cardiovascular diseases. OBJECTIVE: To determine the expression of tryptophan-5-hydroxylase (TPH) 1 and 2, serotonin transporter protein (SERT) and serotonergic receptors in hearts with dilated cardiomyopathy (DCM) compared to controls. METHOD: A comparative study was performed in six tissue blocks of the left ventricular free wall (LVWL) and inter-ventricular septum from patients who died of DCM and six who died of no cardiovascular diseases (controls). Five slices from each block were obtained to determine the expression of TPH1 and TPH2, SERT and serotonergic receptors with antibodies specific for immunofluorescence. Immunofluorescence was analyzed by Student's t-test, accepting a significance level of p < 0.05. RESULTS: An increase in TPH1, TPH2, 5-HT2A and 5-HT2B receptors expression were observed in dilated structures compared to controls (p < 0.05). For dilated inter-ventricular septum, the 5-HT4 receptor increased its expression (p < 0.05), and SERT in PLVI compared to controls (p < 0.05). CONCLUSIONS: These results suggest that the increases observed in the expression of TPH, SERT, and serotonergic receptors in hearts with DCM compared to controls could play an important role in the pathophysiology of MCD in humans.

ANTECEDENTES: Los cardiomiocitos sintetizan, utilizan y recapturan serotonina, la cual participa en la modulación parácrina y autócrina de la actividad del corazón y en la fisiopatología de algunas enfermedades cardiovasculares. OBJETIVO: Determinar la expresión de triptófano-5-hidroxilasa (TPH) 1 y 2, transportador de serotonina (SERT) y receptores serotoninérgicos en corazones con miocardiopatía dilatada (MCD) en comparación con controles. MÉTODO: Estudio comparativo en seis bloques de la pared libre del ventrículo izquierdo (PLVI) y del septum interventricular de pacientes fallecidos por MCD y seis que murieron por enfermedades no cardiovasculares. Se obtuvieron cinco cortes de cada bloque para determinar la expresión de TPH1 y TPH2, SERT y receptores serotoninérgicos con anticuerpos específicos por inmunofluorescencia. La inmunofluorescencia fue analizada por la t de Student, aceptando un nivel de significancia de p < 0.05. RESULTADOS: Se observó un aumento en la expresión de TPH1 y TPH2 y en los receptores 5-HT2A y 5-HT2B en las estructuras dilatadas en comparación con las controles (p < 0.05). El receptor 5-HT4 aumentó su expresión en el septum interventricular dilatado (p < 0.05) y el SERT en la PLVI en comparación con los controles (p < 0.05). CONCLUSIONES: Estos resultados sugieren que los aumentos observados en las expresiones de TPH, SERT y receptores serotoninérgicos en corazones con MCD en comparación con controles podrían desempeñar un papel importante en la fisiopatología de la MCD en los humanos.

La miocardiopatía hipertrófica induce cambios en la expresión de la triptófano-5-hidroxilasa, el transportador de serotonina y receptores serotoninérgicos / Hypertrophic cardiomyopathy induces changes in the tryptophan-5-hydroxylase, serotonin transporter and serotonergic receptors expressions

Resumen Introducción: Los cardiomiocitos poseen la maquinaria bioquímica capaz de sintetizar, utilizar y recapturar serotonina. Objetivo: Determinar si la miocardiopatía hipertrófica (MCH) induce cambios en la expresión de la triptófano-5-hidroxilasa (TPH) 1 y 2, el transportador de serotonina (SERT) y los receptores serotoninérgicos (RS). Métodos: Estudio transversal de cinco bloques de tejido de corazones con MCH y cinco bloques de corazones de control. Se obtuvieron cinco cortes de la pared libre del ventrículo izquierdo (PLVI) y del septum interventricular (SIV) de cada bloque, para determinar la expresión de TPH1 y TPH2, SERT y RS con anticuerpos por inmunofluorescencia. La inmunofluorescencia fue evaluada mediante t de WELCH, con nivel de significación de p < 0.05. Resultados: La PLVI y el SIV de los corazones con MCH mostraron aumento de la expresión de TPH1 y TPH2, así como de los receptores 5-HT2A y 5-HT2B en comparación con los controles (p < 0.01). El receptor 5-HT4 y SERT aumentaron en el SIV de los corazones con MCH (p < 0.01). Conclusiones: Se demostró aumento de las expresiones de TPH, SERT y RS en los cardiomiocitos de los corazones con MCH en comparación con los controles, lo cual podría participar en la fisiopatología de la MCH en los humanos.

Abstract Introduction: Cardiomyocytes have a biochemical machinery with the capacity to synthesize, utilize and reuptake serotonin. Objective: To determine whether hypertrophic cardiomyopathy (HCM) induces changes in the expression of tryptophan-5-hydroxylase (TPH) 1 and 2, serotonin transporter (SERT) and serotonergic receptors (SR). Methods: Cross-sectional study of five tissue blocks from hearts with HCM and five controls. Five sections of the left ventricular free wall (LVFW) and interventricular septum (IVS) were obtained from each block to determine the expression of TPH1 and TPH2, SERT and SRs by immunofluorescence with specific antibodies. Immunofluorescence was evaluated by WELCH t-test, with a level of significance of p < 0.05. Results: LVFW and IVS of hearts with HCM showed an increase in the expression of TPH1 and TPH 2 and 5-HT2A and 5-HT2B receptors in comparison with controls (p < 0.01). The 5-HT4 receptor and SERT showed an increase in the IVS of hearts with HCM (p < 0.01). Conclusions: This study demonstrated an increased expression of TPH, SERT and SRs in cardiomyocytes from hearts with HCM in comparison with controls, which could be involved in the pathophysiology of HCM in humans.

Diabetes mellitus causes changes in the activity and expression of brain tryptophan-5-hydroxylases and in the number of serotonergic neurons, which do not return to normal with insulin treatment.

INTRODUCTION: Diabetes mellitus (DM) inhibits brain serotonin biosynthesis through changes in tryptophan-5-hydroxylase (TPH) activity and expression. OBJECTIVES: To determine whether DM-induced changes in brain TPH1 or TPH2 expression and in the number of serotonergic neurons return to normal in diabetic rats treated with insulin. METHODS: Rats with streptozotocin-induced diabetes were divided in two groups: one treated with insulin and the other without treatment. On day 14, brain stems were obtained in order to quantify L-tryptophan and 5-hydroxytryptamine levels, as well as to determine TPH activity. The expression of TPH1 and TPH2 by West-ern blot, and the number of serotonergic neurons by immunohistochemistry. RESULTS: In diabetic rats, a decrease in the levels of L-tryptophan, 5-hydroxytryptamine, and TPH activity was confirmed, as well as lower TPH1 and TPH2 expression and lower numbers of serotonergic neurons. When diabetic rats were treated with insulin, L-tryptophan returned to normal, but not 5-hy-droxytryptamine, TPH expression, or the number of serotonergic neurons. CONCLUSIONS: DM chronically inhibits the synthesis of brain 5-hydroxytryptamine through changes in TPH1 and TPH2 expression and a decrease in the number of serotonergic neurons, which persist despite insulin treatment.

INTRODUCCIÓN: La diabetes mellitus (DM) inhibe la biosíntesis de serotonina cerebral mediante cambios en la actividad y expresión de la triptófano-5-hidroxilasa (TPH). OBJETIVOS: Determinar si los cambios en la expresión de TPH1 o TPH2 cerebral y en el número de neuronas serotoninérgicas causados por la DM retornan a la normalidad en las ratas con diabetes tratadas con insulina. MÉTODOS: Ratas con diabetes inducida con estreptozotocina se dividieron en dos grupos: uno tratado con insulina y otro sin tratamiento. En el día 14, se obtuvieron tallos cerebrales para cuantificar niveles de L-triptófano, 5-hidroxitriptamina y la actividad de la TPH. La expresión de TPH1 y TPH2 fue mediante Western blot y el número de neuronas serotoninérgicas por inmu­nohistoquímica. RESULTADOS: En las ratas con diabetes se confirmó disminución de los niveles de L-triptófano, 5-hidroxitriptamina y la actividad de la TPH, así como una menor expresión de TPH1 y 2 y un menor número de neuronas serotoninérgicas. Cuando las ratas diabéticas fueron tratadas con insulina, el L-triptófano regreso a la normalidad, no así la 5-hidroxitriptamina, la expresión de TPH y el número de neuronas serotoninérgicas. CONCLUSIONES: La DM inhibe crónicamente la síntesis de 5-hidroxitriptamina cerebral mediante modificaciones en la expresión de TPH1 y TPH2 y disminución de las neuronas seroto­ninérgicas, que persisten a pesar del tratamiento con insulina.

Cambios en la actividad y expresión de las triptófano-5-hidroxilasas cerebrales y en el número de neuronas serotoninérgicas por la diabetes mellitus, que no retornan a la normalidad con la insulina / Diabetes mellitus causes changes in brain tryptophan-5-hydroxylases activity and expression and in the number of serotonergic neurons, which do not return to normal with insulin treatment

Resumen Introducción: La diabetes mellitus (DM) inhibe la biosíntesis de serotonina cerebral mediante cambios en la actividad y expresión de triptófano-5-hidroxilasa (TPH). Objetivos: Determinar si los cambios en la expresión de TPH1 y TPH2 cerebral y en el número de neuronas serotoninérgicas causados por la DM retornan a la normalidad en ratas con diabetes tratadas con insulina. Métodos: Ratas con diabetes inducida con estreptozotocina se dividieron en dos grupos uno tratado con insulina y otro sin tratamiento. En el día 14, se obtuvieron tallos cerebrales para cuantificar niveles de L-triptófano, 5-hidroxitriptamina y la actividad de la TPH. La expresión de TPH1 y TPH2 fue mediante Western blot y el número de neuronas serotoninérgicas por inmunohistoquímica. Resultados: En las ratas con diabetes se confirmó disminución de los niveles de L-triptófano, 5-hidroxitriptamina y la actividad de la TPH, así como menor expresión de TPH1 y TPH2 y menor número de neuronas serotoninérgicas. Cuando las ratas diabéticas fueron tratadas con insulina, el L-triptófano regresó a la normalidad, no así la 5-hidroxitriptamina, la expresión de TPH ni el número de neuronas serotoninérgicas. Conclusiones: La DM inhibe crónicamente la síntesis de 5-hidroxitriptamina cerebral mediante modificaciones en la expresión de TPH1 y TPH2 y disminución de las neuronas serotoninérgicas, que persisten a pesar del tratamiento con insulina.

Abstract Introduction: Diabetes mellitus (DM) inhibits brain serotonin biosynthesis through changes in tryptophan-5-hydroxylase (TPH) activity and expression. Objectives: To determine whether DM-induced changes in brain TPH1 and TPH2 expression and in the number of serotonergic neurons return to normal in diabetic rats treated with insulin. Methods: Rats with streptozotocin-induced diabetes were divided in two groups: one treated with insulin and the other without treatment. On day 14, brain stems were obtained in order to quantify L-tryptophan and 5-hydroxytryptamine levels, as well as to determine TPH activity. The expressión of TPH1 and THP2 by Western blot, and the number of serotonergic neurons by immunohistochemistry. Results: In diabetic rats, a decrease in the levels of L-tryptophan, 5-hydroxytryptamine and TPH activity was confirmed, as well as lower TPH1 and TPH2 expression and lower numbers of serotonergic neurons. When diabetic rats were treated with insulin, L-tryptophan returned to normal, but not 5-hydroxytryptamine, TPH expression, or the number of serotonergic neurons. Conclusions: DM chronically inhibits the synthesis of brain 5-hydroxytryptamine through changes in TPH1 and TPH2 expression and a decrease in the number of serotonergic neurons, which persist despite insulin treatment.

Hypertrophic cardiomyopathy induces changes in the tryptophan-5-hydroxylase, serotonin transporter and serotonergic receptors expressions.

INTRODUCTION: Cardiomyocytes have a biochemical machinery with the capacity to synthesize, utilize and reuptake serotonin. OBJECTIVE: To determine whether hypertrophic cardiomyopathy (HCM) induces changes in the expression of tryptophan-5-hydroxylase (TPH) 1 and 2, serotonin transporter (SERT) and serotonergic receptors (SR). METHODS: Cross-sectional study of five tissue blocks from hearts with HCM and five controls. Five sections of the left ventricular free wall (LVFW) and interventricular septum (IVS) were obtained from each block to determine the expression of TPH1 and TPH2, SERT and SRs by immunofluorescence with specific antibodies. Immunofluorescence was evaluated by WELCH t-test, with a level of significance of p < 0.05. RESULTS: LVFW and IVS of hearts with HCM showed an increase in the expression of TPH1 and TPH 2 and 5-HT2A and 5-HT2B receptors in comparison with controls (p < 0.01). The 5-HT4 receptor and SERT showed an increase in the IVS of hearts with HCM (p < 0.01). CONCLUSIONS: This study demonstrated an increased expression of TPH, SERT and SRs in cardiomyocytes from hearts with HCM in comparison with controls, which could be involved in the pathophysiology of HCM in humans.

INTRODUCCIÓN: Los cardiomiocitos poseen la maquinaria bioquímica capaz de sintetizar, utilizar y recapturar serotonina. OBJETIVO: Determinar si la miocardiopatía hipertrófica (MCH) induce cambios en la expresión de la triptófano-5-hidroxilasa (TPH) 1 y 2, el transportador de serotonina (SERT) y los receptores serotoninérgicos (RS). MÉTODOS: Estudio transversal de cinco bloques de tejido de corazones con MCH y cinco bloques de corazones de control. Se obtuvieron cinco cortes de la pared libre del ventrículo izquierdo (PLVI) y del septum interventricular (SIV) de cada bloque, para determinar la expresión de TPH1 y TPH2, SERT y RS con anticuerpos por inmunofluorescencia. La inmunofluorescencia fue evaluada mediante t de WELCH, con nivel de significación de p < 0.05. RESULTADOS: La PLVI y el SIV de los corazones con MCH mostraron aumento de la expresión de TPH1 y TPH2, así como de los receptores 5-HT2A y 5-HT2B en comparación con los controles (p < 0.01). El receptor 5-HT4 y SERT aumentaron en el SIV de los corazones con MCH (p < 0.01). CONCLUSIONES: Se demostró aumento de las expresiones de TPH, SERT y RS en los cardiomiocitos de los corazones con MCH en comparación con los controles, lo cual podría participar en la fisiopatología de la MCH en los humanos.

Desenvolvimento de bebidas lácteas probióticas adicionadas de polpa de morango e suplementadas com L-triptofano elaboradas a partir de leite bubalino e bovino / Development of probiotic dairy beverages with strawberry pulp and supplemented with L-tryptophan from buffalo and bovine milk

O objetivo deste estudo foi desenvolver uma formulação de bebida láctea bubalina probiótica adicionada de polpa de morango, comparando os efeitos do uso do leite de búfala e de vaca na elaboração dos produtos e verificando a possibilidade de suplementação com triptofano nos produtos lácteos probióticos. Como primeira etapa do trabalho, bebidas lácteas probióticas foram elaboradas a partir de leite bubalino e bovino, fermentadas com Streptococcus thermophilus TA040, Lactobacillus bulgaricus LB340 e Lactobacillus acidophilus La5, e formuladas com 0, 25 e 50% de soro em sua formulação. As bebidas foram avaliadas quanto à cinética de fermentação das culturas láticas utilizadas, ao teor de proteína, gordura e sólidos totais não gordurosos, pós-acidificação, viabilidade das culturas fermentadoras e sua capacidade de sobrevivência ao estresse gastrointestinal in vitro. As bebidas lácteas bubalinas apresentaram resultados superiores as bebidas bovinas. O uso do leite de búfala na elaboração das bebidas lácteas promoveu benefícios quanto as culturas láticas presentes nos produtos, exercendo efeito protetivo e influindo na preservação da viabilidade das bactérias ao longo do armazenamento refrigerado e durante a simulação do estresse gastrointestinal in vitro. As bebidas lácteas elaboradas com 25% apresentaram os resultados mais próximos aos obtidos pelos produtos controle, sem adição de soro, sendo selecionadas para a segunda parte do estudo. Nesta etapa, as formulações de bebida láctea com 25% de soro, foram acrescidas de um preparado com polpa de morango e bebidas sem adição da fruta, utilizadas como controle. As bebidas lácteas bubalinas frutadas, apresentaram menor teor de gordura e melhores características reológicas, com maior viscosidade e consistência do que os produtos controle, sem afetar a pós-acidificação, o perfil de ácido graxo, assim como, a viabilidade e a resistência às condições de estresse gastrointestinal in vitro das culturas fermentadoras. A avaliação da possibilidade de suplementar lácteos probióticos com triptofano foi realizada em conjunto com a Universidade de Milão. Para isso, iogurtes probióticos receberam adição de triptofano antes ou após a fermentação, sendo avaliados com relação ao perfil de pós-acidificação, quantidade de triptofano nos produtos, número de células viáveis por plaqueamento e citometria de fluxo ao longo do armazenamento a 25° e 4°C. Complementarmente, a influência da presença do triptofano no crescimento e produção de compostos antimicrobianos pelas culturas láticas, também foi avaliada. A adição de triptofano após a fermentação dos iogurtes, que foram armazenados sob refrigeração (4°C), além de não afetar a pós-acidificação dos produtos, apresentou benefícios quanto a viabilidade L. acidophilus, redução do dano e aumento do número de células vivas, promovendo teor maior do aminoácido nos iogurtes. A presença do triptofano nos meios de cultivo, também influenciou de forma positiva o crescimento de S. thermophilus e L. acidophilus, melhorando o desenvolvimento das bactérias durante a fermentação e influindo em uma maior atividade antilistérica por parte do S. thermophilus. Diante da influência positiva da aplicação do leite de búfala na elaboração das bebidas lácteas, assim como, a adição do triptofano em iogurtes probióticos, a suplementação do aminoácido em bebidas lácteas bubalinas frutadas permitiria a obtenção de um produto funcional, onde seus benefícios estariam relacionados tanto ao consumo do probiótico presente no produto quanto a complementação de triptofano na dieta do consumidor

The aim of this study was to develop a formulation of probiotic buffalo dairy beverage added with strawberry pulp, comparing the effects of using buffalo and cow's milk in the preparation of products and verifying the possibility of tryptophan supplementation in probiotic dairy products. As a first stage of the work, probiotic dairy beverages were made from buffalo and bovine milk, fermented with Streptococcus thermophiles TA040, Lactobacillus bulgaricus LB340 and Lactobacillus acidophilus La5, and formulated with 0, 25 and 50% whey in their formulation. The beverages were evaluated for the fermentation kinetics of the used lactic cultures, the levels of protein, fat and total no fat solids, post-acidification, fermenting cultures viability and their ability to survive gastrointestinal stress in vitro. Buffalo milk use in dairy beverages production promoted benefits regarding the lactic cultures present in the products, exerting a protective effect and influencing the viability preservation of bacteria during the cold storage and simulation of gastrointestinal stress in vitro. Dairy beverages made with 25% whey addition showed results similar to those obtained by the control products, without whey addition, being selected for the second part of the study. In this part, the dairy beverages formulations with 25% whey, were added with a preparation were added with a strawberry pulp preparation and dairy beverages without added fruit, used as a control. Fruity bubaline dairy beverages had lower fat content and better rheological characteristics, with higher viscosity and consistency than control products, without affecting post-acidification, fatty acid profile, as well as viability and resistance to in vitro gastrointestinal condition of fermented cultures. The possibility of supplementing probiotic dairy products with tryptophan was evaluated in partnership with the University of Milan. For this, probiotic yogurts received the addition of tryptophan before or after fermentation, being evaluated in relation to the post-acidification profile, tryptophan amount in the products, viable cell number per plating and flow cytometry during storage at 25°C and 4°C. In addition, the influence of the tryptophan presence on the growth and production of antimicrobial compounds by lactic cultures was also evaluated. The addition of tryptophan after the yogurt fermentation, which were stored under refrigeration (4°C), in addition to not affecting the post-acidification of the products, showed benefits to the viability of L. acidophilus, reduced the damage and increased the number of cells promoting higher amino acid content in yogurts. Tryptophan presence in the culture media also positively influenced the growth of S. thermophiles and L. acidophilus, improving the development of bacteria during fermentation and influencing better antilisteric activity in the part of S. thermophiles. In view of the buffalo milk positive influence observed after the application in dairy beverage preparation, as well as the addition of tryptophan in probiotic yoghurts, amino acid supplementation in fruity buffalo dairy beverages would allow to obtain a functional product, where its benefits would be related both to the consumption of the probiotic present in the product as to the supplementation of tryptophan in the consumer's diet

Desenvolvimento de bebidas lácteas probióticas adicionadas de polpa de morango e suplementadas com L-triptofano elaboradas a partir de leite bubalino e bovino / Development of probiotic dairy beverages with strawberry pulp and supplemented with L-tryptophan from buffalo and bovine milk

O objetivo deste estudo foi desenvolver uma formulação de bebida láctea bubalina probiótica adicionada de polpa de morango, comparando os efeitos do uso do leite de búfala e de vaca na elaboração dos produtos e verificando a possibilidade de suplementação com triptofano nos produtos lácteos probióticos. Como primeira etapa do trabalho, bebidas lácteas probióticas foram elaboradas a partir de leite bubalino e bovino, fermentadas com Streptococcus thermophilus TA040, Lactobacillus bulgaricus LB340 e Lactobacillus acidophilus La5, e formuladas com 0, 25 e 50% de soro em sua formulação. As bebidas foram avaliadas quanto à cinética de fermentação das culturas láticas utilizadas, ao teor de proteína, gordura e sólidos totais não gordurosos, pós-acidificação, viabilidade das culturas fermentadoras e sua capacidade de sobrevivência ao estresse gastrointestinal in vitro. As bebidas lácteas bubalinas apresentaram resultados superiores as bebidas bovinas. O uso do leite de búfala na elaboração das bebidas lácteas promoveu benefícios quanto as culturas láticas presentes nos produtos, exercendo efeito protetivo e influindo na preservação da viabilidade das bactérias ao longo do armazenamento refrigerado e durante a simulação do estresse gastrointestinal in vitro. As bebidas lácteas elaboradas com 25% apresentaram os resultados mais próximos aos obtidos pelos produtos controle, sem adição de soro, sendo selecionadas para a segunda parte do estudo. Nesta etapa, as formulações de bebida láctea com 25% de soro, foram acrescidas de um preparado com polpa de morango e bebidas sem adição da fruta, utilizadas como controle. As bebidas lácteas bubalinas frutadas, apresentaram menor teor de gordura e melhores características reológicas, com maior viscosidade e consistência do que os produtos controle, sem afetar a pós-acidificação, o perfil de ácido graxo, assim como, a viabilidade e a resistência às condições de estresse gastrointestinal in vitro das culturas fermentadoras. A avaliação da possibilidade de suplementar lácteos probióticos com triptofano foi realizada em conjunto com a Universidade de Milão. Para isso, iogurtes probióticos receberam adição de triptofano antes ou após a fermentação, sendo avaliados com relação ao perfil de pós-acidificação, quantidade de triptofano nos produtos, número de células viáveis por plaqueamento e citometria de fluxo ao longo do armazenamento a 25° e 4°C. Complementarmente, a influência da presença do triptofano no crescimento e produção de compostos antimicrobianos pelas culturas láticas, também foi avaliada. A adição de triptofano após a fermentação dos iogurtes, que foram armazenados sob refrigeração (4°C), além de não afetar a pós-acidificação dos produtos, apresentou benefícios quanto a viabilidade L. acidophilus, redução do dano e aumento do número de células vivas, promovendo teor maior do aminoácido nos iogurtes. A presença do triptofano nos meios de cultivo, também influenciou de forma positiva o crescimento de S. thermophilus e L. acidophilus, melhorando o desenvolvimento das bactérias durante a fermentação e influindo em uma maior atividade antilistérica por parte do S. thermophilus. Diante da influência positiva da aplicação do leite de búfala na elaboração das bebidas lácteas, assim como, a adição do triptofano em iogurtes probióticos, a suplementação do aminoácido em bebidas lácteas bubalinas frutadas permitiria a obtenção de um produto funcional, onde seus benefícios estariam relacionados tanto ao consumo do probiótico presente no produto quanto a complementação de triptofano na dieta do consumidor

The aim of this study was to develop a formulation of probiotic buffalo dairy beverage added with strawberry pulp, comparing the effects of using buffalo and cow's milk in the preparation of products and verifying the possibility of tryptophan supplementation in probiotic dairy products. As a first stage of the work, probiotic dairy beverages were made from buffalo and bovine milk, fermented with Streptococcus thermophiles TA040, Lactobacillus bulgaricus LB340 and Lactobacillus acidophilus La5, and formulated with 0, 25 and 50% whey in their formulation. The beverages were evaluated for the fermentation kinetics of the used lactic cultures, the levels of protein, fat and total no fat solids, post-acidification, fermenting cultures viability and their ability to survive gastrointestinal stress in vitro. Buffalo milk use in dairy beverages production promoted benefits regarding the lactic cultures present in the products, exerting a protective effect and influencing the viability preservation of bacteria during the cold storage and simulation of gastrointestinal stress in vitro. Dairy beverages made with 25% whey addition showed results similar to those obtained by the control products, without whey addition, being selected for the second part of the study. In this part, the dairy beverages formulations with 25% whey, were added with a preparation were added with a strawberry pulp preparation and dairy beverages without added fruit, used as a control. Fruity bubaline dairy beverages had lower fat content and better rheological characteristics, with higher viscosity and consistency than control products, without affecting post-acidification, fatty acid profile, as well as viability and resistance to in vitro gastrointestinal condition of fermented cultures. The possibility of supplementing probiotic dairy products with tryptophan was evaluated in partnership with the University of Milan. For this, probiotic yogurts received the addition of tryptophan before or after fermentation, being evaluated in relation to the post-acidification profile, tryptophan amount in the products, viable cell number per plating and flow cytometry during storage at 25°C and 4°C. In addition, the influence of the tryptophan presence on the growth and production of antimicrobial compounds by lactic cultures was also evaluated. The addition of tryptophan after the yogurt fermentation, which were stored under refrigeration (4°C), in addition to not affecting the post-acidification of the products, showed benefits to the viability of L. acidophilus, reduced the damage and increased the number of cells promoting higher amino acid content in yogurts. Tryptophan presence in the culture media also positively influenced the growth of S. thermophiles and L. acidophilus, improving the development of bacteria during fermentation and influencing better antilisteric activity in the part of S. thermophiles. In view of the buffalo milk positive influence observed after the application in dairy beverage preparation, as well as the addition of tryptophan in probiotic yoghurts, amino acid supplementation in fruity buffalo dairy beverages would allow to obtain a functional product, where its benefits would be related both to the consumption of the probiotic present in the product as to the supplementation of tryptophan in the consumer's diet

How intrauterine growth restriction due to nutritional stress changes the function of key proteins in brain serotonin metabolism during development.

This review aimed to describe and comment on how experimental intrauterine nutritional stress in animals produced some changes in tryptophan-5-hydroxylases (TPH) 1 and 2 in the brain and other key proteins such as plasma albumin, and how the intrauterine nutritional stress could produce long-lasting alterations in serotonin function in the brain of human infants.

El objetivo de esta revisión es describir y comentar cómo el estrés nutricional intrauterino experimental en animales produjo algunos cambios en las triptófano-5-hidroxilasas 1 y 2 en el cerebro y en otras proteínas clave, como la albúmina plasmática, y de qué manera el estrés nutricional intrauterino podría producir alteraciones duraderas en la función de la serotonina en el cerebro de lactantes.

A via das quinureninas e suas implicações na esquizofrenia / Kynurenine pathway and its implications in schizophrenia

Introdução: a esquizofrenia é uma doença mental grave heterogênea, de extrema complexidade, manifestada por vários distúrbios de cognição, pensamento, memória, comportamento e afeto. Sua fisiopatologia, apesar dos avanços, continua de difícil compreensão. Objetivo: realizar uma revisão da literatura a respeito da relação entre a via das quinureninas e a fisiopatologia da esquizofrenia. Metodologia: foi conduzida a busca de artigos através das bases de dados Medline e Lilacs, utilizando os descritores schizophrenia/ esquizofrenia e as palavras-chave kynurenine pathways/via das quinureninas. Por meio desta busca, foram selecionados 38 artigos publicados entre os anos de 1990 e 2018. Resultados: estudos mais recentes vêm demonstrando que anormalidades no metabolismo do triptofano por meio da via das quinureninas poderiam estar relacionadas com os mecanismos neurofisiopatológicos da esquizofrenia. Os metabólitos gerados pela via das quinureninas possuem propriedades neuroativas. Dentre eles, destaca-se o ácido quinurênico, antagonista endógeno do receptor N-metil-D-aspartato. Seus níveis cerebrais estão aumentados em pacientes esquizofrênicos, provavelmente devido a uma condição pró-inflamatória, resultando dessa forma, em possíveis alterações neurológicas responsáveis pelo desenvolvimento da esquizofrenia. Conclusão: não somente uma melhor compreensão da fisiopatologia da esquizofrenia, como também o desenvolvimento de novos alvos terapêuticos, poderão ser obtidos a partir de um melhor entendimento da relação entre anormalidades na via das quinureninas e a gênese da esquizofrenia.

Introduction: Schizophrenia is a severe heterogeneous mental illness of extreme complexity, manifested by various disorders of cognition, thought, memory, behavior and affection. Its pathophysiology, despite advances, remains difficult to understand. Objective: to review the literature on the relationship between the kynurenine pathway and the pathophysiology of schizophrenia. Methodology: a literature review was carried out using the descriptors kynurenine pathways/via das quinureninas and the keywords kynurenine pathways/via das quinureninas using Medline and Lilacs databases. Through this search, were selected 38 articles published between the years of 1990 and 2018. Results: recent studies have shown that abnormalities in tryptophan metabolism through the kynurenine pathway may be related to the neuropathophysiological mechanisms of schizophrenia. The metabolites generated by the kynurenine pathway have neuroactive properties. Among them, kynurenic acid, an endogenous antagonist of the N-methyl-D-aspartate receptor, stands out. Their brain levels are increased in schizophrenic patients, probably due to a pro-inflammatory condition, resulting in possible neurological changes responsible for the development of schizophrenia. Conclusion: not only a better knowledge of pathophysiology, but also the development of new therapeutic targets will be obtained with a better understanding of the relationship between abnormalities in the kynurenine pathway and the genesis of schizophrenia.

Sinalização e metabolismo celulares durante a diferenciação de linfócitos B humanos em secretores de anticorpos na presença do vírus da Dengue in vitro e em pacientes com Dengue / B cell signaling and metabolism during antibody-secreting cell differentiation in the presence of Dengue virus in vitro and in Dengue patients

A Dengue é uma doença viral sistêmica, transmitida por mosquitos, que afeta anualmente cerca de 100 milhões de pessoas em todo o mundo. Causada por quatro sorotipos do vírus da Dengue (DENV), suas manifestações clínicas podem variar de assintomáticas à formas que podem levar a óbito. Curiosamente, os pacientes com Dengue apresentam uma resposta exacerbada das células secretoras de anticorpos (ASCs) no sangue cerca de sete dias após o início dos sintomas. A frequência dessas ASCs induzidas pelo DENV representa mais de 50% de todas as células B circulantes no sangue. Essa quantificação é maior que aquelas encontradas em outras infecções virais, contextos de imunização e até mesmo em pacientes com neoplasias de ASCs. Além disso, a magnitude dessa resposta transitória se correlaciona com a gravidade da doença. Então, como a infecção pelo DENV induz essa resposta enorme? Para responder à essa pergunta, combinamos abordagens in vitro e in silico. Células mononucleares do sangue periférico (PBMCs) obtidas de indivíduos saudáveis foram cultivadas in vitro durante sete dias na presença do DENV ou mitógenos. Após a estimulação pelo DENV, as células B presentes nas PBMCs foram capazes de se diferenciarem em ASCs, tanto fenotipicamente quanto funcionalmente, em magnitude similar àquelas estimuladas com mitógenos. Essa diferenciação demonstrou ser dependente da presença de outras células contidas nas PBMCs, assim como do contato célula-célula. Embora ambos os estímulos tenham sido capazes de induzir a diferenciação de ASCs, eles diferiram metabolicamente e transcricionalmente. PBMCs estimuladas pelo DENV apresentaram um maior consumo de triptofano, associado à maior expressão de IDO1 e IDO2 e maior síntese de quinurenina, bem como maiores expressões de IL-10, BAFF e SYK. Ainda, as concentrações de quinurenina foram positivamente correlacionadas com a enumeração de ASCs nessas culturas. Dados de transcriptoma públicos de pacientes com Dengue também suportam esses achados. Outros flavivírus, como o vírus Zika e a cepa vacinal da Febre Amarela não foram capazes de induzir a mesma magnitude de diferenciação das células B em ASCs in vitro. Tão pouco apresentaram correlação entre a enumeração de ASCs e a síntese de quinurenina. Por fim, através da construção de uma hipotética via de diferenciação de células B em ASCs durante infecção pelo DENV, através da combinação de dados da literatura e transcriptomas públicos, demonstramos que moléculas relacionadas à via do STAT3 (IL-10, IL-6, IRF4 e BLIMP1) estão mais expressas nos pacientes infectados e moléculas que respondem aos sinais de cálcio (Calcineurina, NFATC1, DOK3 e GRB2) estão menos expressas nos pacientes infectados. Esses dados proporcionam um melhor entendimento da resposta de células B durante a infeção pelo DENV, particularmente sobre como o metabolismo e a sinalização das células B estão conectados nesse processo

Dengue is a mosquito-borne viral disease that affects annually about 100 million people worldwide. Caused by four Dengue virus (DENV) serotypes, it ranges from asymptomatic to life threatening forms. Curiously, Dengue patients present an exacerbated blood antibody-secreting cell (ASCs) response around seven days after the symptoms onset. The frequency of those DENV-induced ASCs represents more than 50% of all circulating blood B cells. This is greater than found in others viral infections, immunization contexts and even in ASCs related-leukemia patients. Moreover, the magnitude of that transitory response correlates with the disease severity. So, how does the DENV infection induce this enormous response? In order to answer this question we have combined in vitro and in silico approaches. Peripheral blood mononuclear cells (PBMC) obtained from healthy individuals were cultured in vitro during seven days in the presence of DENV or mitogens. Upon the DENV stimulation, PBMC-contained B cells were able to differentiate phenotypically and functionally into ASCs, both phenotypically and functionally, in a similar magnitude than mitogen-stimulated cells. This differentiation was demonstrated to be dependent of the presence of the remaining PBMCs, as well as of the cell-cell contact. Although both stimuli were able to induce the ASCs differentiation, they differed metabolically and transcriptionally. DENV-stimulated PBMCs showed higher tryptophan consumption, associated with higher IDO1 and IDO2 expression and higher kynurenine synthesis, as well as higher IL-10, BAFF and SYK expressions compared to mitogen-exposed counterparts. Additionally, the kynurenine concentrations were positively correlated with the ASCs-enumeration in those cultures. Public transcriptome data supports these findings as well. Other flaviviruses, such as Zika virus and the attenuated vaccine Yellow Fever were not able to induce the same magnitude of ASCs differentiation in vitro. Hence, they did not present a correlation between the number of generated ASCs and the supernatant kynurenine levels. Based on the combination of the literature and public transcriptome data, we have constructed a hypothetical B cell differentiation pathway that might be occurring during DENV infection. It displays that STAT3 pathway-related molecules (IL-10, IL-6, IRF4 and BLIMP1) are more expressed in Dengue patients and molecules that respond to calcium signals (Calcineurin, NFATC1, DOK3 and GRB2) are less expressed in Dengue patients than in control. These data provide a better understanding of the B cell response elicited by DENV infection, particularly about how the B cell metabolism and signaling can be connected into this process

Acute and chronic bovine pulmonary edema and emphysema in Uruguay / Edema e enfisema pulmonar agudo e crônico em bovinos do Uruguai

An outbreak of pulmonary edema and emphysema with acute and chronic cases is reported in a farm in Uruguay. In a herd of 40 Hereford steers, 20 died. The deaths began four days after a change of paddock, from an old pasture of Avena sativa to a lush growing pasture of the same grass. Acutely affected animals showed severe dyspnea, sialorrhea, cough, and subcutaneous edema, and died within 72 hours. Chronically affected steers showed dyspnea, respiratory noises, weight loss, and intolerance to exercise. The deaths began four days after the change of paddock. Ten days after the first death, the steers were withdrawn from the pasture, but continued dying throughout the following 40 days. Twenty animals died and six were necropsied. Grossly, the lungs were diffusely armed and glistening, with reddish and crepitant cut surface, and presented alveolar septae sharply distended by edema and emphysema. There was subpleural emphysema with air blebs distributed across the pleural surface. Presence of Dictyocaulus viviparus was observed in three steers. In some animals, the trachea was diffusely reddish with presence of pink foam; in some others, there was bloody liquid in the tracheal lumen. Histologic examination showed severe diffuse alveolar and interstitial emphysema, hyaline membranes adhered to the alveolar wall, thickening of the interlobular septae with proliferation of type II pneumocytes, and moderate-to-severe multifocal histiocytic, neutrophilic and eosinophilic infiltrate. In the trachea, there was submucosal hemorrhage and moderate multifocal eosinophilic and lymphocytic infiltrate. The steers with chronic signs presented similar lung lesions, but multifocal pulmonary fibrosis and cardiac dilatation were also observed. The diagnosis of acute bovine pulmonary emphysema and edema (ABPE) was based on the occurrence of the disease after introduction of the herd in a lush green pasture, on the characteristic gross and histologic lesions, and on the absence of other toxic or infectious agents causing similar lesions. Cattle raisers should be alert to the risks of occurrence of this disease after the introduction of the herds into paddocks with green and lush pastures.(AU)

Descreve-se um surto de edema e enfisema pulmonar com casos agudos e crônicos em bovinos em uma criação semi-intensiva no Uruguai. De um lote de 40 novilhos da raça Hereford morreram 20. As mortes começaram quatro dias após uma mudança de alimentação, de uma pastagem mais velha de Avena sativa, para uma pastagem recentemente plantada de aveia que estava em brotação. Os animais afetados apresentaram sinais clínicos agudos de dispneia, sialorreia, tosse e alguns desenvolveram edema subcutâneo, morrendo em até 72 horas. Outros novilhos mais cronicamente afetados apresentaram dispneia, ruídos respiratórios, perda de peso e intolerância ao exercício. As mortes começaram quatro dias após a mudança de pastagens. Dez dias após a primeira morte, os novilhos foram retirados do pasto, mas morreram ainda durante 40 dias mais. Ao total, morreram vinte animais e seis foram necropsiados. Nas necropsias dos animais mortos na fase aguda os pulmões estavam difusamente armados e brilhosos e ao corte de coloração avermelhada e crepitante, com os septos alveolares acentuadamente distendidos por edema e enfisema. Havia enfisema subpleural caracterizado por bolhas de ar distribuídas pela superfície pleural. Em três bovinos havia ainda presença de Dictyocaulus viviparus. Alguns animais apresentaram a traqueia difusamente avermelhada com espuma de coloração rósea ou liquido sanguinolento livre na luz traqueal. Histologicamente havia edema e enfisema alveolar e intersticial difuso severo, membranas hialinas espessas aderidas à parede alveolar, espessamento dos septos interlobulares com proliferação de pneumócitos tipo II e infiltrado inflamatório histiocítico, neutrofílico e eosinofílico multifocal moderado a severo. Na traqueia havia hemorragias na submucosa e infiltrado eosinofílico e linfocítico multifocal. Os novilhos com sinais crônicos apresentaram lesões pulmonares semelhantes, entretanto, foram observadas também, fibrose pulmonar multifocal e dilatação cardíaca. O diagnóstico de EEPAB baseou-se na ocorrência da doença após a introdução do rebanho em uma pastagem viçosa em brotação, nas características macroscópicas e histológicas e na ausência de outros agentes tóxicos ou infecciosos que causam lesões semelhantes. Se alerta para os riscos da ocorrência desta enfermidade, quando houver mudanças de pastagens.(AU)

Expressão de indoleamina 2,3-dioxigenase (IDO) e triptofano 2,3-dioxigenase(TDO) no ambiente cervicovaginal normal, na vaginose bacteriana e nas lesões cervicais associadas ao HPV / Expression of indoleamine 2,3-dioxygenase (IDO) and tryptophan 2,3-dioxygenase (TDO) in normal cervicovaginal environment, bacterial vaginosis and cervical lesions associated with HPV

Neste estudo avaliamos o papel do metabolismo do triptofano (Trp) na homeostasia, na vaginose bacteriana e nas lesões cervicais associadas ao HPV. A importância do metabolismo do Trp se deve a sua ação na proliferação de microrganismos e de células do sistema imune. O consumo de triptofano tem sido identificado como uma forma de controlar o crescimento bacteriano limitando a infecção. Por outro lado, a oxidação de Trp produz quinurenina (QUIN), que tem papel chave na tolerância imunológica. A formação de QUIN se dá através das enzimas indoleamina 2,3-dioxigenase (IDO) e triptofano 2,3- dioxigenase (TDO). A mais estudada delas no âmbito das infecções/ imuno escape é a enzima IDO. Mais recentemente, tem-se dado ênfase ao papel da TDO no câncer. Nesta dissertação, o interesse foi avaliar a expressão da IDO no epitélio cervicovaginal de mulheres com vaginose bacteriana e de IDO e TDO em amostras cervicais de mulheres com diferentes graus de lesão cervical associada ao HPV. Foram incluídas 165 mulheres atendidas no CAISM/UNICAMP, as quais foram divididas em dois grupos: grupo caso composto por mulheres com lesão de baixo ou alto grau e carcinoma invasor (n=42) e grupo controle composto por mulheres com citologia oncológica normal, independente de apresentar infecção genital (n=123). IDO foi avaliada por imunocitoquímica em citologia em base líquida e IDO e TDO em biópsias cervicais. Mulheres com vaginose bacteriana apresentaram expressão aumentada de IDO em células escamosas em comparação às mulheres sem vaginose bacteriana (OR=7.41; IC 95%= 2.50 a 21.4; p <0.0001). No epitélio vaginal normal com ou sem infecção por HPV houve uma expressão leve de IDO em células escamosas. Na presença de lesões ou carcinoma, houve um aumento no número de células escamosas displásicas e de leucócitos IDO-positivos; aumento de IDO também pôde ser observada em culturas de pele organotípicas transduzidas com as oncoproteínas E6/ E7 do HPV16. Nas lesões cervicais, assim como visto para a IDO, a TDO esteve expressa em leucócitos, especialmente os infiltrados na região estromal e na parede dos vasos sanguíneos. A expressão basal de IDO no epitélio cervical normal e sua regulação positiva na infecção por HPV e lesões associadas sugerem a participação do metabolismo do Trp nos mecanismos imunossupressores envolvidos na doença. Embora o papel do IDO já tenha sido abordada anteriormente, até onde sabemos esta é a primeira evidência da expressão de TDO no epitélio vaginal, na neoplasia intraepitelial cervical e carcinoma de células escamosas. Ainda, em leucócitos, especialmente aqueles com morfologia típica de polimorfonucleares, parecem ser importantes fontes de IDO na cérvix uterina

In this study we evaluated the role of tryptophan (Trp) metabolism in cervix homeostasis, bacterial vaginosis and HPV-associated lesions. The importance of Trp metabolism is due to its action on microorganisms and immune cells. Tryptophan consumption has been identified as a way to controlling bacterial growth limiting infection. On the other hand, the oxidation of Trp produces kynurenine (Kyn) which plays a key role in immunological tolerance. The formation of Kyn occurs through the enzymes indoleamine 2,3-dioxygenase (IDO) and tryptophan 2,3-dioxygenase (TDO). IDO is the most studied of them within the context of infections / immune escape. More recently, TDO has also been considered in studies of cancer progression. In this thesis, we were interested in cervicovaginal epithelium IDO expression in women with bacterial vaginosis and of IDO and TDO in cervical samples of women with different degrees of cervical lesion associated with HPV. A total of 165 women attended at CAISM/UNICAMP were divided into two groups: a case group composed of women with low or high grade lesions and invasive carcinoma (n = 42) and a control group composed of women with normal cytology, independent to present genital infection (n =123). IDO was evaluated by immunocytochemistry in liquid-based cytology and IDO and TDO in cervical biopsies. Women with bacterial vaginosis had increased IDO expression in squamous cells compared to women without bacterial vaginosis (OR = 7.41, 95% CI = 2.50- 21.74; p<0.0001). In normal vaginal epithelium with or without HPV infection there was a mild IDO expression in squamous cells. In the presence of cervical intraepithelial lesions or squamous cell carcinoma, there was an increase in the number of IDO-positive dysplastic squamous cells and leukocytes; increase in IDO can also be observed in organotypic skin cultures transduced with HPV-16 E6/E7 oncoproteins. In cervical lesions, as observed for IDO, TDO was expressed in leukocytes, especially infiltrates in the stromal region and in the wall of blood vessels. The basal expression of IDO in the normal cervical epithelium and its positive regulation in HPV infection and associated lesions suggests the participation of Trp metabolism in the immunosuppressive mechanisms involved in the disease. Although some previous data have already considered the role of IDO, as far as we know this is the first evidence of the participation of TDO in the vaginal epithelium, cervical intraepithelial neoplasia and squamous cell carcinoma. In addition, in leukocytes, especially those with a typical polymorphonuclear morphology, appear to be important sources of IDO in the uterine cervix

Characterisation of three polymorphisms of the tryptophan hydroxylase 2 gene in a sample of Colombian population with major depressive disorder / Caracterización de tres polimorfismos del gen de la triptófano hidroxilasa 2 en una muestra de población colombiana con trastorno depresivo mayor

ABSTRACT Objective: Identify whether rs11179000, rs136494 and rs4570625 polymorphisms of the tryptophan hydroxylase 2 gene, are associated with a major depressive disorder in a sample of the Colombian population. Methods: Case-control study was conducted in which a comparison was made between subjects diagnosed with major depressive disorder at some point in adulthood or active symptoms at the time of evaluation, and subjects with no psychiatric disease. Subjects were studied in the Department of Psychiatry, Faculty of Medicine and the Institute of Genetics at the National University of Colombia. Polymorphisms were genotyped using Taqman probes in real time PCR. As well as studying the association between major depressive disorder and these single nucleotide polymorphisms (SNPs), the association with other factors previously associated with depression were also analysed. Results: No statistically significant association between genotypic and allelic frequencies of each polymorphism and major depressive disorder was found. Association between sex and complication during pregnancy/childbirth and major depressive disorder was observed. Association between sex and complication during pregnancy/childbirth and major depres sive disorder was observed. Conclusions: There was no association between any polymorphism and major depressive disorder.

RESUMEN Objetivo: Identificar si los polimorfismos rs11179000, rs136494 y rs4570625 del gen de la triptófano hidroxilasa 2 están asociados a trastorno depresivo mayor en una muestra de población colombiana. Métodos: Estudio de casos y controles en el que se comparó a sujetos con trastorno depresivo mayor diagnosticado en algún momento de la vida adulta o con síntomas activos en el momento de la valoración y sujetos sin enfermedad psiquiátrica. Se estudió a los sujetos en el Departamento de Psiquiatría de la Facultad de Medicina y en el Instituto de Genética de la Universidad Nacional de Colombia. Se genotipificaron los polimorfismos usando reacción en cadena de la polimerasa en tiempo real y sondas Taqman. Además de buscar asociación entre trastorno depresivo mayor y estos polimorfismos de un solo nucleótido, se exploró asociación con otros factores relacionados previamente con depresión. Resultados: No se encontró asociación estadísticamente significativa entre las frecuencias genotípicas o alélicas de cada polimorfismo y el trastorno depresivo mayor. Se observó asociación entre sexo y complicaciones durante el embarazo/parto y trastorno depresivo mayor. Conclusiones: No se halló asociación entre polimorfismo alguno y el trastorno depresivo mayor.

[Characterisation of three polymorphisms of the tryptophan hydroxylase 2 gene in a sample of Colombian population with major depressive disorder]. / Caracterización de tres polimorfismos del gen de la triptófano hidroxilasa 2 en una muestra de población colombiana con trastorno depresivo mayor.

OBJECTIVE: Identify whether rs11179000, rs136494 and rs4570625 polymorphisms of the tryptophan hydroxylase 2 gene, are associated with a major depressive disorder in a sample of the Colombian population. METHODS: Case-control study was conducted in which a comparison was made between subjects diagnosed with major depressive disorder at some point in adulthood or active symptoms at the time of evaluation, and subjects with no psychiatric disease. Subjects were studied in the Department of Psychiatry, Faculty of Medicine and the Institute of Genetics at the National University of Colombia. Polymorphisms were genotyped using Taqman probes in real time PCR. As well as studying the association between major depressive disorder and these (single nucleotide polymorphisms (SNPs), the association with other factors previously associated with depression were also analysed. RESULTS: No statistically significant association between genotypic and allelic frequencies of each polymorphism and major depressive disorder was found. Association between sex and complication during pregnancy / childbirth and major depressive disorder was observed. Association between sex and complication during pregnancy / childbirth and major depressive disorder was observed. CONCLUSIONS: There was no association between any polymorphism and major depressive disorder.

Study of l-tryptophan in an experimental model of depression caused by alzheimer's disease / Estudo do l-triptofano na depressão ocorrida pela doença de alzheimer em modelo experimental

ABSTRACT Alzheimer's is a neurodegenerative disease characterized by cognitive impairment commonly associated with mood disorders, which trigger depressive reactions and compromise mental performance and functionality. The objective of the present study was to verify the effects of L-tryptophan and to analyze motor behavior in an experimental model of depression caused by the Alzheimer's disease process. The sample consisted of 40 Wistar rats divided equally into two groups, 20 animals treated with L-tryptophan and 20 control animals. Both groups received spatial memory training in the water maze task and were submitted to stereotaxic surgery to induce dementia. The treated group achieved better spatial memory activity than the control group in the Morris water maze. Treatment with L-tryptophan had beneficial effects on reactive memory.

RESUMO O Alzheimer é uma doença neurodegenerativa caracterizado pelo comprometimento cognitivo comumente associado a transtornos do humor, os quais desencadeiam reações depressivas, comprometem o desempenho mental e a funcionalidade. O objetivo do presente estudo foi verificar os efeitos do L-triptofano e analisar o comportamento motor em modelo experimental com depressão decorrente do processo de Alzheimer. A amostra foi composta por 40 ratos da linhagem wistar divididos igualmente em dois grupos, 20 animais tratados com L-triptofano e 20 animais pertencentes ao grupo controle. Ambos os grupos receberam treinamento da memória espacial no later water maze e foram submetidos à cirurgia estereotáxica para indução demencial. Verificou-se através do labirinto aquático de Morris que o grupo tratado obteve atividade para memória espacial melhor do que o grupo controle. O tratamento com L-triptofano demonstrou melhor benefício na memória reativa.

Is there a relationship between tryptophan dietary intake and plasma levels of indoxyl sulfate in chronic kidney disease patients on hemodialysis? / Há relação entre ingestão alimentar de triptofano e níveis plasmáticos de indoxil sulfato em pacientes renais crônicos em hemodiálise?

Abstract Introduction: Gut microbiota is involved in generation of uremic toxins in chronic kidney disease (CKD) patients on hemodialysis (HD), like indoxyl sulfate (IS) that is originated from tryptophan amino acid fermentation. Objective: To evaluate the tryptophan intake by chronic renal failure patients on HD and its possible relationship with IS plasma levels. Methods: Participated of the study 46 patients with CKD on HD regular program (56.5% men; 52.7 ± 10.3 years; 63 (32.2-118.2) months on HD; BMI 25.6 ± 4.9 kg/m2). The tryptophan intake was evaluated by a 24-hours dietary recall (R-24h) performed on 3 different days. Routine biochemical tests and anthropometric measurements were evaluated. IS plasma levels were determined by High Performance Liquid Chromatography (HPLC) with fluorescent detection and the interleukin-6 (IL-6) plasma levels by immunoenzymatic method (ELISA, Enzyme Linked Immunosorbent Assay). Results: The average of tryptophan intake was according to recommendation, but IS plasma levels (35.0 ± 11.9 mg/L) were elevated, however according to the EUTox values for uremic individuals. There was no correlation between the tryptophan intake and IS plasma levels. However, there was positive correlation between protein intake and tryptophan and variables used to evaluate lean body mass, and moreover, IS levels were positively associated with IL-6 (r = 0.6: p = 0.01). Conclusion: The present study suggests that tryptophan dietary intake may not be a determinant factor to IS levels. However, it suggests that gut microbiota may play an important role in systemic inflammation in patients with CKD.

Resumo Introdução: A microbiota intestinal está envolvida na geração de toxinas urêmicas presentes nos pacientes com doença renal crônica (DRC) em hemodiálise (HD) como indoxil sulfato (IS), formado a partir da fermentação do aminoácido triptofano. Objetivo: Avaliar a ingestão de triptofano alimentar pelos pacientes renais crônicos em HD e sua possível relação com os níveis plasmáticos de IS. Métodos: Participaram do estudo 46 pacientes com DRC em programa regular de HD (56,5% homens; 52,7 ± 10,3 anos; 63 (32,2-118,2) meses em HD; IMC 25,6 ± 4,9kg/m2. A ingestão de triptofano foi avaliada por meio do recordatório alimentar de 24 (R-24h) realizado em três diferentes dias. Exames bioquímicos de rotina, bem como a avaliação antropométrica foram avaliados. Os níveis plasmáticos de IS foram determinados por cromatografia líquida de alto desempenho (HPLC) com detecção fluorescente e as concentrações plasmáticas de interleucina-6 (IL-6) pelo método imunoenzimático (ELISA, Enzyme Linked Immunosorbent Assay). Resultados: A ingestão média de triptofano estava dentro do recomendado, já os níveis plasmáticos de IS (35,0 ± 11,9mg/L) estavam elevados, porém de acordo com os valores da EUTox para indivíduos urêmicos. Não houve correlação entre a ingestão de triptofano e os níveis plasmáticos de IS. Contudo, houve correlação positiva entre ingestão de proteína e triptofano e variáveis que avaliam massa magra e, além disso, os níveis IS foram positivamente associados com os de IL-6 (r = 0,6: p = 0,01). Conclusão: O presente estudo sugere que a ingestão alimentar de triptofano pode não ser um fator determinante dos níveis de IS. No entanto, sugere que o intestino pode ter importante papel na inflamação sistêmica presente nos pacientes com DRC.