Introdução: Wistar Audiogenic Rats (WAR) é modelo experimental desenvolvido na Faculdade de Medicina de Ribeirão Preto, para estudo da epilepsia, entretanto, a seleção genética em resposta aos comportamentos de crises audiogênicas também trouxe à tona alterações no metabolismo energético nesses animais. Dois estudos são relevantes deste ponto de vista, no primeiro Botion e Doretto observaram que WAR após serem estimulados apresentam: (a) valores de glicemia maior em relação ao Wistar; (b) aumento no lactato circulante - o que pode indicar deficiência na fosforilação oxidativa (OXPHOS); (c) aumento da atividade adrenérgica, induzindo dessensibilização da via lipolítica ?-adrenérgica no tecido adiposo epididimal. Pereira e colaboradores investigando o metabolismo de carboidratos relataram: (a) aumento na via glicólica; (b) translocação de GLUT4 aumentada no músculo gastrocnêmico e (c) redução nos níveis de glicogênio muscular. Objetivo: Diante desses achados o presente estudo tem como objetivo elucidar o metabolismo energético mitocondrial e sua relação com as crises epiléticas induzidas por estímulos sonoros. Resultados: Através de analises comportamentais e calorimetria indireta, relatamos que WAR possui perfil ansiogênico e tem preferência em oxidar aminoácidos; utilizando biopsias de tecido hepático, musculo esquelético e cardíaco, observamos maior densidade mitocondrial, acompanhada de maior geração de H2O2 e como consequência maior estresse oxidativo; na busca para elucidar com maior destreza, realizamos isolamento da fração mitocondrial do tecido hepático, e concluímos que não há maior geração de H2O2 por mitocôndria, porém há um enriquecimento proteico (algumas proteínas funcionais - como as envolvidas na OXPHOS- e outras não-funcionais - como as UCPs), além de relatarmos maiores níveis de proteínas envolvidas na dinâmica mitocondrial, desse modo podemos concluir que WAR possuem maior densidade mitocondrial e mitocôndrias com maior eficiência; o mesmo perfil mitocondrial foi observado no tecido cerebral. Após tratamento com DNP e NAC, observamos redução do estresse oxidativo no tecido cerebral, porém apenas NAC reduziu a severidade da crise, assim concluímos que o suave desacoplamento induzido por DNP não é capaz de reduzir de forma significativa a severidade da crise, porém a inibição da glicólise pelo NAC, alterou a bioenergética cerebral é capaz a reduzir a severidade da crise, deixando evidenciado que o metabolismo energético tem papel essencial/relevante na crise epilética induzida por estímulos sonoros em WAR, enquanto o estresse oxidativo tem papel secundário. / Introduction: Wistar Audiogenic Rats (WAR) is an experimental model developed in the Ribeirão Preto Medical School, for the study of epilepsy, however, the genetic selection in response to the behaviors of audiogenic crisis also brought up changes in energy metabolism in these animals. Two studies are relevant from this point of view, in the first Botion and Doretto observed that WAR after being stimulated present: (a) higher blood glucose values in relation to the Wistar; (b) Increase in circulating lactatewhich may indicate deficiency in oxidative phosphorylation (OXPHOS); (c) Increased adrenergic activity. Pereira and collaborators investigating the metabolism of carbohydrates reported: (a) increase in glycolysis; (b) Translocation of GLUT4 increased in gastrocnemius muscle and (c) reduction in muscle glycogen levels. Objective: In the face of these findings, the present study aims to elucidate the mitochondrial energy metabolism and its relation to the epileptic crises induced by sound stimuli. Results: Through behavioral analysis and indirect calorimetry, we report that WAR has reduced exploratory activity and has preference to oxidize amino acids; Using biopsies of liver tissue, skeletal and cardiac muscles, we observe greater mitochondrial density, accompanied by greater generation of H2O2 and as a result of greater oxidative stress; in the search to elucidate with greater dexterity, we perform isolation of the mitochondrial fraction of the hepatic tissue, and we conclude that there is no greater generation of H2O2 by mitochondria, but there is a protein enrichment (some functional proteins - such as those involved in OXPHOS - and other nonfunctional ones - such as UCPs), in addition to reporting higher levels of proteins involved in mitochondrial dynamics, so we can conclude that WAR has greater mitochondrial density and mitochondria more efficiently; the same mitochondrial profile was observed in the brain tissue. After treatment with DNP and NAC, we observed reduction of oxidative stress in the brain tissue, but only NAC reduced the severity of the crisis, so we conclude that the smooth decoupling induced by DNP is not able to significantly reduce the severity of the crisis, however the inhibition of glycolysis by the NAC, altered the brain bioenergetics is able to reduce the severity of the crisis, leaving evidence that the energy metabolism has essential/relevant role in the epileptic crisis induced by sound stimuli in WAR, while oxidative stress has secondary role.

INTRODUÇÃO: Metionina (Met), cisteína (Cys), homocisteína (Hcy) e taurina (Tau) são os quatro aminoácidos sulfurados (AAS), mas apenas a Met e Cys são incorporadas em proteínas. Os três principais produtos doS AAS, glutationa, (GSH), Hcy e Tau influenciam, principalmente, as respostas inflamatória e imune. A Tau e GSH diminuem a inflamação, enquanto que a Hcy apresenta efeito oposto. Os pacientes HIV+ apresentam baixos níveis de GSH e outros nutrientes antioxidantes, mostrando relação direta entre Cys (e GSH) com células CD4+. Não se conhece o mecanismo pelo qual as mudanças na ingestão dos AAS influenciam este fenômeno. Paralelamente, as relações entre Hcy, doenças inflamatórias e alterações in vitro no comportamento das células imunes levantou ressalvas sobre a suplementação de dietas com AAS. OBJETIVOS : investigar as vias dos AAS em pacientes HIV+ nas condições de jejum e pós-sobrecarga de Met frente à dieta habitual (OH) isolada ou acompanhada da suplementação de Cys (NAC) ou glutamina (Gln). MÉTODOS : 12 pacientes HIV+ (6 M e 6 F, de 25 a 36 anos), sob tratamento anti-retroviral pelo esquema tríplice, sem infecções secundárias e 20 controles saudáveis (10M e 10F, 23-28 anos) foram randomicamente distribuídos para suplementação com NAC (N-acetilcisteína, 1g/d) ou Gln (20 g/d) em estudo cruzado com 7 dias de dieta separados por uma semana de washout (Wo com DH). Amostras de sangue após jejum noturno de 10 a 12 horas foram coletadas antes (MO) e após (M1) cada regime dietético. A seguir, os indivíduos ingeriram metionina (100 mg/kg), com coletas de sangue após 2 e 4 horas para a determinação da área abaixo da curva (AAC). No MO, ambos os grupos foram avaliados quanto à antropometria (IMC, kg/m2), funções glomerular (uréia, creatinina) e hepatocelular (γ-GT), estados nutricional (albumina, cálcio, ácido fólico e vitamina 812) e antioxidante (ácido úrico, GSH, GSSG, Hcy), glicose, lipídios (triacilgliceróis e frações de colesterol) e AAS, serina (Ser), glicina (Gly), glutamato (Glu) e Gln. O grupo HIV também foi caracterizado pela carga viral e contagem de CD4+ e CD8+. As comparações estatísticas entre os grupos e entre as dietas mostraram homogeneidade para IMC, albumina, cálcio, vitamina 812, Hcy, HDL-colesterol, uréia e creatinina. Os pacientes apresentaram valores maiores de glicose, triacilgliceróis, γ-GT, LDL-colesterol e GSSG paralelalemente às menores concentrações de ácido úrico, GSH e todos os AAS, exceto Hcy. A sobrecarga de metionina igualou (pelos valores de delta) os grupos para Met, Hcy, Tau e Gln. As suplementações de NAC e Gln levaram o grupo HIV+ a concentrações maiores de GSH (NAC > Gln), atuando diferentemente em seus precursores: G/y (Gln > NAC) e Cys (NAC > Gln) e resultando em consumo similar de Ser e produção de Tau. Ambas as dietas reduziram GSSG/GSH (NAC > Gln) e apenas NAC aumentou (6 x) a Hcy. Esta última foi piorada pela sobrecarga de Mel. Assim, HIV+ resulta em deficiências múltiplas de vitaminas e aminoácidos levando a menores níveis de GSH e GSSG/GSH mais elevada. Os principais problemas de menor formação de Cys e menor incorporação de Cys em GSH foram resolvidos dando-se Met, NAC e Gln aos pacientes, ainda permanecendo a desvantagem do aumento da Hcy com Met ou suplementação de NAC. / BACKGROUNO: Methionine (Met), cysteine (Cys), homocysteine (Hcy), and, taurine (Tau) are the 4 sulfur-containing amino acids (SAA), but only Met and Cys are incorporated into proteins. The 3 major products of SAA, glutathione (GSH), Hcy and Tau influence, mainly, inflammatory and of immune responses. Tau and GSH ameliorate inflammation whereas Hcy has the opposite effect. HIV+ patients present low levelis of GSH and other antioxidants nutrients, showing a direct relationship between Cys (and GSH) with CD4+/ cells. How changes in SAA intake influence this phenomenon is unknown and the relationships among Hcy, inflammatory diseases, and in vitro alterations in immune cell behavior create a cautionary note about supplementation of diets with SAA. OBJECTIVE: To investigate SAA pathways in HIV+ patients on fast and Met-overload (Met-DL) states after taken diet habitual without (HD) or with supplements of Cys (NAC) or glutamine (Gln). METHOOS: 12 HIV+ (6M and 6F, 25-36 yrs old) patients under HAART without secondary infections and 20 healthy (10M and 10F, 23-28 yrs old) controls were randomly assigned to either NAC (N-acetylcysteine, 1g/d) or Gln (20g/d) diets, in a 7-day diet crossover design, separated by a 7-day washout (with HD) period. Blood samples were drawn after overnight fast before (MO) and after each dietary treatments (M1) for the resting measurements. Immediately after blood sampling ali subjects started the Met-DL by ingesting at once 100 mg Met/kg BW and having the blood draw after 2 and 4 hours for the area under the curve (AUC) determination. At MO both groups were assessed for anthropometry (BMI, kg/m2), glomerular (plasma urea and creatinina) and hepatocellular (plasma γGT activity) funetions, nutritional (albumin, calcium, folic acid and vitamin B12) and antioxidant (uric acid, GSH, GSSG, Hey) states, glucose, lipids (triglycerides and cholesterol fractions) and SAA, serine (Ser), glyeine (Gly), glutamate (Glu) and Gln. The HIV+ group was characterized also by viral load, CD4+ and CD8+ counts. The statistical comparisons between groups and among diets showed group homogeneity for 8MI, albumin, calcium, vitamin B12, Hey, HDL-cholesterol, urea and creatinine. The patients presented higher values of glucose, triglycerides, γ-GT, LDL-cholesterol, and GSSG along with lower concentrations of uric acid, GSH and all but Hcy amino acids. The Met-OL equalized (Δ values) the groups for Met, Hcy, Tau and Gln. NAC and Gln diets led the HIV+ group to a higher concentrations of GSH (NAC > Gln) by acting differently on its precursors: Gly (Gln > NAC) and Cys (NAC > Gln), resulting similar consumption of Ser and production of Tau. Both diets reduced GSSG/GSH (NAC > Gln) and only NAC increased (6 x) Hey. The later was worsened by Met-OL. Thus HIV+ results in multiple deficiencies of vitamins and amino acids leading to lower levels of GSH and higher GSSG/GSH ration. The main problems of lower formation of Cys and low ineorporation of Cys and Gly into GSH were greatly solved by giving Met, NAC and Gln to the patients, hence remaining the drawback of increasing Hcy with Met or NAC supplements.

Introdução: Wistar Audiogenic Rats (WAR) é modelo experimental desenvolvido na Faculdade de Medicina de Ribeirão Preto, para estudo da epilepsia, entretanto, a seleção genética em resposta aos comportamentos de crises audiogênicas também trouxe à tona alterações no metabolismo energético nesses animais. Dois estudos são relevantes deste ponto de vista, no primeiro Botion e Doretto observaram que WAR após serem estimulados apresentam: (a) valores de glicemia maior em relação ao Wistar; (b) aumento no lactato circulante - o que pode indicar deficiência na fosforilação oxidativa (OXPHOS); (c) aumento da atividade adrenérgica, induzindo dessensibilização da via lipolítica ?-adrenérgica no tecido adiposo epididimal. Pereira e colaboradores investigando o metabolismo de carboidratos relataram: (a) aumento na via glicólica; (b) translocação de GLUT4 aumentada no músculo gastrocnêmico e (c) redução nos níveis de glicogênio muscular. Objetivo: Diante desses achados o presente estudo tem como objetivo elucidar o metabolismo energético mitocondrial e sua relação com as crises epiléticas induzidas por estímulos sonoros. Resultados: Através de analises comportamentais e calorimetria indireta, relatamos que WAR possui perfil ansiogênico e tem preferência em oxidar aminoácidos; utilizando biopsias de tecido hepático, musculo esquelético e cardíaco, observamos maior densidade mitocondrial, acompanhada de maior geração de H2O2 e como consequência maior estresse oxidativo; na busca para elucidar com maior destreza, realizamos isolamento da fração mitocondrial do tecido hepático, e concluímos que não há maior geração de H2O2 por mitocôndria, porém há um enriquecimento proteico (algumas proteínas funcionais - como as envolvidas na OXPHOS- e outras não-funcionais - como as UCPs), além de relatarmos maiores níveis de proteínas envolvidas na dinâmica mitocondrial, desse modo podemos concluir que WAR possuem maior densidade mitocondrial e mitocôndrias com maior eficiência; o mesmo perfil mitocondrial foi observado no tecido cerebral. Após tratamento com DNP e NAC, observamos redução do estresse oxidativo no tecido cerebral, porém apenas NAC reduziu a severidade da crise, assim concluímos que o suave desacoplamento induzido por DNP não é capaz de reduzir de forma significativa a severidade da crise, porém a inibição da glicólise pelo NAC, alterou a bioenergética cerebral é capaz a reduzir a severidade da crise, deixando evidenciado que o metabolismo energético tem papel essencial/relevante na crise epilética induzida por estímulos sonoros em WAR, enquanto o estresse oxidativo tem papel secundário. / Introduction: Wistar Audiogenic Rats (WAR) is an experimental model developed in the Ribeirão Preto Medical School, for the study of epilepsy, however, the genetic selection in response to the behaviors of audiogenic crisis also brought up changes in energy metabolism in these animals. Two studies are relevant from this point of view, in the first Botion and Doretto observed that WAR after being stimulated present: (a) higher blood glucose values in relation to the Wistar; (b) Increase in circulating lactatewhich may indicate deficiency in oxidative phosphorylation (OXPHOS); (c) Increased adrenergic activity. Pereira and collaborators investigating the metabolism of carbohydrates reported: (a) increase in glycolysis; (b) Translocation of GLUT4 increased in gastrocnemius muscle and (c) reduction in muscle glycogen levels. Objective: In the face of these findings, the present study aims to elucidate the mitochondrial energy metabolism and its relation to the epileptic crises induced by sound stimuli. Results: Through behavioral analysis and indirect calorimetry, we report that WAR has reduced exploratory activity and has preference to oxidize amino acids; Using biopsies of liver tissue, skeletal and cardiac muscles, we observe greater mitochondrial density, accompanied by greater generation of H2O2 and as a result of greater oxidative stress; in the search to elucidate with greater dexterity, we perform isolation of the mitochondrial fraction of the hepatic tissue, and we conclude that there is no greater generation of H2O2 by mitochondria, but there is a protein enrichment (some functional proteins - such as those involved in OXPHOS - and other nonfunctional ones - such as UCPs), in addition to reporting higher levels of proteins involved in mitochondrial dynamics, so we can conclude that WAR has greater mitochondrial density and mitochondria more efficiently; the same mitochondrial profile was observed in the brain tissue. After treatment with DNP and NAC, we observed reduction of oxidative stress in the brain tissue, but only NAC reduced the severity of the crisis, so we conclude that the smooth decoupling induced by DNP is not able to significantly reduce the severity of the crisis, however the inhibition of glycolysis by the NAC, altered the brain bioenergetics is able to reduce the severity of the crisis, leaving evidence that the energy metabolism has essential/relevant role in the epileptic crisis induced by sound stimuli in WAR, while oxidative stress has secondary role.

INTRODUÇÃO: Metionina (Met), cisteína (Cys), homocisteína (Hcy) e taurina (Tau) são os quatro aminoácidos sulfurados (AAS), mas apenas a Met e Cys são incorporadas em proteínas. Os três principais produtos doS AAS, glutationa, (GSH), Hcy e Tau influenciam, principalmente, as respostas inflamatória e imune. A Tau e GSH diminuem a inflamação, enquanto que a Hcy apresenta efeito oposto. Os pacientes HIV+ apresentam baixos níveis de GSH e outros nutrientes antioxidantes, mostrando relação direta entre Cys (e GSH) com células CD4+. Não se conhece o mecanismo pelo qual as mudanças na ingestão dos AAS influenciam este fenômeno. Paralelamente, as relações entre Hcy, doenças inflamatórias e alterações in vitro no comportamento das células imunes levantou ressalvas sobre a suplementação de dietas com AAS. OBJETIVOS : investigar as vias dos AAS em pacientes HIV+ nas condições de jejum e pós-sobrecarga de Met frente à dieta habitual (OH) isolada ou acompanhada da suplementação de Cys (NAC) ou glutamina (Gln). MÉTODOS : 12 pacientes HIV+ (6 M e 6 F, de 25 a 36 anos), sob tratamento anti-retroviral pelo esquema tríplice, sem infecções secundárias e 20 controles saudáveis (10M e 10F, 23-28 anos) foram randomicamente distribuídos para suplementação com NAC (N-acetilcisteína, 1g/d) ou Gln (20 g/d) em estudo cruzado com 7 dias de dieta separados por uma semana de washout (Wo com DH). Amostras de sangue após jejum noturno de 10 a 12 horas foram coletadas antes (MO) e após (M1) cada regime dietético. A seguir, os indivíduos ingeriram metionina (100 mg/kg), com coletas de sangue após 2 e 4 horas para a determinação da área abaixo da curva (AAC). No MO, ambos os grupos foram avaliados quanto à antropometria (IMC, kg/m2), funções glomerular (uréia, creatinina) e hepatocelular (γ-GT), estados nutricional (albumina, cálcio, ácido fólico e vitamina 812) e antioxidante (ácido úrico, GSH, GSSG, Hcy), glicose, lipídios (triacilgliceróis e frações de colesterol) e AAS, serina (Ser), glicina (Gly), glutamato (Glu) e Gln. O grupo HIV também foi caracterizado pela carga viral e contagem de CD4+ e CD8+. As comparações estatísticas entre os grupos e entre as dietas mostraram homogeneidade para IMC, albumina, cálcio, vitamina 812, Hcy, HDL-colesterol, uréia e creatinina. Os pacientes apresentaram valores maiores de glicose, triacilgliceróis, γ-GT, LDL-colesterol e GSSG paralelalemente às menores concentrações de ácido úrico, GSH e todos os AAS, exceto Hcy. A sobrecarga de metionina igualou (pelos valores de delta) os grupos para Met, Hcy, Tau e Gln. As suplementações de NAC e Gln levaram o grupo HIV+ a concentrações maiores de GSH (NAC > Gln), atuando diferentemente em seus precursores: G/y (Gln > NAC) e Cys (NAC > Gln) e resultando em consumo similar de Ser e produção de Tau. Ambas as dietas reduziram GSSG/GSH (NAC > Gln) e apenas NAC aumentou (6 x) a Hcy. Esta última foi piorada pela sobrecarga de Mel. Assim, HIV+ resulta em deficiências múltiplas de vitaminas e aminoácidos levando a menores níveis de GSH e GSSG/GSH mais elevada. Os principais problemas de menor formação de Cys e menor incorporação de Cys em GSH foram resolvidos dando-se Met, NAC e Gln aos pacientes, ainda permanecendo a desvantagem do aumento da Hcy com Met ou suplementação de NAC. / BACKGROUNO: Methionine (Met), cysteine (Cys), homocysteine (Hcy), and, taurine (Tau) are the 4 sulfur-containing amino acids (SAA), but only Met and Cys are incorporated into proteins. The 3 major products of SAA, glutathione (GSH), Hcy and Tau influence, mainly, inflammatory and of immune responses. Tau and GSH ameliorate inflammation whereas Hcy has the opposite effect. HIV+ patients present low levelis of GSH and other antioxidants nutrients, showing a direct relationship between Cys (and GSH) with CD4+/ cells. How changes in SAA intake influence this phenomenon is unknown and the relationships among Hcy, inflammatory diseases, and in vitro alterations in immune cell behavior create a cautionary note about supplementation of diets with SAA. OBJECTIVE: To investigate SAA pathways in HIV+ patients on fast and Met-overload (Met-DL) states after taken diet habitual without (HD) or with supplements of Cys (NAC) or glutamine (Gln). METHOOS: 12 HIV+ (6M and 6F, 25-36 yrs old) patients under HAART without secondary infections and 20 healthy (10M and 10F, 23-28 yrs old) controls were randomly assigned to either NAC (N-acetylcysteine, 1g/d) or Gln (20g/d) diets, in a 7-day diet crossover design, separated by a 7-day washout (with HD) period. Blood samples were drawn after overnight fast before (MO) and after each dietary treatments (M1) for the resting measurements. Immediately after blood sampling ali subjects started the Met-DL by ingesting at once 100 mg Met/kg BW and having the blood draw after 2 and 4 hours for the area under the curve (AUC) determination. At MO both groups were assessed for anthropometry (BMI, kg/m2), glomerular (plasma urea and creatinina) and hepatocellular (plasma γGT activity) funetions, nutritional (albumin, calcium, folic acid and vitamin B12) and antioxidant (uric acid, GSH, GSSG, Hey) states, glucose, lipids (triglycerides and cholesterol fractions) and SAA, serine (Ser), glyeine (Gly), glutamate (Glu) and Gln. The HIV+ group was characterized also by viral load, CD4+ and CD8+ counts. The statistical comparisons between groups and among diets showed group homogeneity for 8MI, albumin, calcium, vitamin B12, Hey, HDL-cholesterol, urea and creatinine. The patients presented higher values of glucose, triglycerides, γ-GT, LDL-cholesterol, and GSSG along with lower concentrations of uric acid, GSH and all but Hcy amino acids. The Met-OL equalized (Δ values) the groups for Met, Hcy, Tau and Gln. NAC and Gln diets led the HIV+ group to a higher concentrations of GSH (NAC > Gln) by acting differently on its precursors: Gly (Gln > NAC) and Cys (NAC > Gln), resulting similar consumption of Ser and production of Tau. Both diets reduced GSSG/GSH (NAC > Gln) and only NAC increased (6 x) Hey. The later was worsened by Met-OL. Thus HIV+ results in multiple deficiencies of vitamins and amino acids leading to lower levels of GSH and higher GSSG/GSH ration. The main problems of lower formation of Cys and low ineorporation of Cys and Gly into GSH were greatly solved by giving Met, NAC and Gln to the patients, hence remaining the drawback of increasing Hcy with Met or NAC supplements.

Introduction. While opioid medications are commonly prescribed for management of neuropathic pain (NP), long-term use has been associated with increased risk for overdose, drug interactions and addiction. New strategies are necessary to better manage chronic pain, thereby reducing need for opioid medications and their associated adverse consequences. N-acetyl-L-cysteine (NAC), an over-the-counter supplement, has shown promise in the treatment of psychiatric and addictive disorders. In addition, NAC has shown promise for reducing physiological signs of NP in laboratory rat models, prompting this study. Purpose. The present study was an open-label clinical trial of NAC as an adjuvant to opioid treatment for poorly controlled, chronic NP. It examined whether 1200 mg NAC twice daily for 4 weeks was associated with: lower ratings of patient-reported pain; reductions in PRN opioid medication for breakthrough pain; and improvements in physical and mental health quality of life (QoL). The study also examined whether appraisal of pain impacts response to medication. Method. Participants were N=28 chronic NP patients who consented to study participation. This consisted of 2 baseline assessments, 4 weeks of NAC and 1 post-trial follow-up visit. The majority (N=17) dropped out or were excluded during baseline. Of the remaining participants, N = 11 started the study medication and N=10 completed the study, with daily recordings of pain severity ratings and use of PRN opioid medication. Small sample size limited analyses to qualitative case reviews and effect sizes. Results. Over 90% of participants receiving NAC completed the study. Case review found varied results. While 4 of 10 participants showed decrease in average pain ratings during NAC, estimated effect sizes for the whole sample were small, bordering on negligible (ω² from .003 to .027) as were those for PRN opioids (Partial Eta-Squared=.0003). Effect size for mental health QoL was medium (Cohen's d=.421). Conclusions. With N=10, findings must be interpreted with caution. Nonetheless, the study found some albeit small evidence supporting NAC for improving mental health QoL and pain ratings. Several participants reported improvements in pain and mental health domains while taking NAC. NAC was well tolerated with minimal side effects. Lessons from this study will inform design and implementation of future NAC studies.

P1 prophage is stably maintained in E. coli as a low-copy-number plasmid. Stable maintenance of P1 is dependent on the function of the plasmid encoded partition system, parABS. ParA is the partition ATPase, ParB is the partition-site binding protein, and parS is the partition site. The concerted action of these proteins results in dynamic movement of the plasmid over the bacterial nucleoid, which results in its stable maintenance. Plasmid movement has been proposed to be caused by interactions between parS bound ParB and nucleoid bound ParA. In this thesis, I have identified a complex of ParA, ParB, and DNA that is capable of promoting plasmid stability. ParA, ParB, DNA interactions required the ATP bound conformation of ParA. The ParA-ParB-DNA complex was dynamically regulated by nucleotide hydrolysis, which promoted complex disassembly. Complex formation resulted from the cooperative binding of ParA and ParB to DNA. ParA-ParB and ParB-DNA interactions were both necessary for complex formation. ParA-ParB-DNA complex size was regulated by ParB stimulation of ParA-ATP hydrolysis. Microscopy demonstrated that complexes resulted in the association of multiple DNA molecules due to protein binding. The properties of complex assembly, dynamics, and DNA grouping lead me to propose a model where associations between ParA bound to the bacterial nucleoid and the partition complex mediated plasmid movement and localization.

P1 prophage is stably maintained in E. coli as a low-copy-number plasmid. Stable maintenance of P1 is dependent on the function of the plasmid encoded partition system, parABS. ParA is the partition ATPase, ParB is the partition-site binding protein, and parS is the partition site. The concerted action of these proteins results in dynamic movement of the plasmid over the bacterial nucleoid, which results in its stable maintenance. Plasmid movement has been proposed to be caused by interactions between parS bound ParB and nucleoid bound ParA. In this thesis, I have identified a complex of ParA, ParB, and DNA that is capable of promoting plasmid stability. ParA, ParB, DNA interactions required the ATP bound conformation of ParA. The ParA-ParB-DNA complex was dynamically regulated by nucleotide hydrolysis, which promoted complex disassembly. Complex formation resulted from the cooperative binding of ParA and ParB to DNA. ParA-ParB and ParB-DNA interactions were both necessary for complex formation. ParA-ParB-DNA complex size was regulated by ParB stimulation of ParA-ATP hydrolysis. Microscopy demonstrated that complexes resulted in the association of multiple DNA molecules due to protein binding. The properties of complex assembly, dynamics, and DNA grouping lead me to propose a model where associations between ParA bound to the bacterial nucleoid and the partition complex mediated plasmid movement and localization.

Made available in DSpace on 2015-04-16T15:23:30Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 11567929 bytes, checksum: 6da47c4eb2763296a1acb028a37478bb (MD5) Previous issue date: 2012-04-28 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / This survey seeks to establish relationships with different knowledge areas, like the Information Science, Visual Arts and the Social Memory, through technical and methodological tools, as well as, from organization models, aimed at treatment of the acquis of the Contemporary Art s Core of Paraíba NAC/UFPB, composed of a variety of informational sources, related to contemporary art. Taking as a basis the discussion about the relationship between information and memory and seeking to understand these phenomena from the study and of the classification and representation of the acquis of NAC. In order to constitute the representation of information with a view to their recovery, through informational signs, aiming a re-meaning of the incorporated memories of the Core throughout its existence. Noting the specific problems and issues brought by the organization of a collection of contemporary art, based on Becker (1997), in instruments of documentary hypothesis analysis, through the informational structures of objects, and still in the goals and perspectives of the Information Science today, the methodological procedures adopted provided structuring a model organization itself, who systematized and presented more dynamically the information contained in the acquis, retrieving them and constituting new meanings to the memory of the NAC/UFPB. / Esta pesquisa busca estabelecer relações com áreas de conhecimentos distintas, como a Ciência da Informação, Artes Visuais e Memória Social, por meio de instrumentos metodológicos e técnicos, bem como, a partir de modelos de organização, visando o tratamento do acervo do Núcleo de Arte Contemporânea da Paraíba NAC/UFPB, composto por uma diversidade de fontes informacionais, ligadas à arte contemporânea. Tendo como base a discussão sobre a relação entre informação e memória e buscando compreender estes fenômenos a partir do estudo e da classificação e representação do acervo do NAC. Com o intuito de constituir a representação das informações com vistas a sua recuperação, através de signos informacionais, visando uma re-significação das memórias constituídas pelo Núcleo, ao longo de sua existência. Observando as especificidades e questões trazidas pela organização de um acervo de arte contemporânea, com base em Becker (1997), nos instrumentos de análise documentária, por meio das estruturas informativas dos objetos, e ainda nos objetivos e perspectivas da Ciência da Informação hoje, os procedimentos metodológicos adotados proporcionaram a estruturação de um modelo próprio de organização, que sistematizou e apresentou de forma mais dinâmica as informações contidas no acervo, recuperando-as e constituindo novos significados à memória do NAC/UFPB.

Alcohol represents the third leading cause of preventable death in the United States. Yet, despite its prevalent role in impeding human health, there is much to understand about how it elicits its effects on the body and how the body and brain change when an individual becomes physiologically dependent upon alcohol. The work presented herein represents an effort to elucidate the acute and chronic effects of alcohol on the nervous system. We investigate two specific protein pathways and their role in alcohol's effects on the body. The first begins with brain-derived neurotrophic factor (BDNF), which acts on TrkB, and ends with KCC2. We demonstrate that BDNF expression is increased in the VTA during withdrawal from chronic but not acute alcohol exposure and that this increase persists for at least seven days. Concomitantly, we demonstrate that the activation of GABAA channels on produces less inhibition of VTA GABA neurons in mice treated with chronic intermittent ethanol exposure than in alcohol naïve mice. This effect likewise persisted for at least seven days. We illustrate that BDNF has no apparent direct effect on VTA GABA neuron firing rate. The second pathway begins with the T cell marker CD5 and ends with the anti-inflammatory cytokine, IL-10. We demonstrate that in a genetic CD5 knockout (CD5 KO) mouse model both alcohol consumption as well as the sedative properties of alcohol are reduced. Since CD+ B cells secrete more IL-10 than CD5- B cells, we also demonstrate the effects of IL-10 on VTA neurons. We show that IL-10 has direct effects on VTA dopamine (DA) neurons by increasing their firing activity. We relatedly illustrate that IL-10 produces an increase in DA release in the nucleus accumbens (NAc). However, contrary to our hypotheses, we show that IL-10 produces conditioned place aversion rather than conditioned place preference in a place conditioning paradigm, suggesting that IL-10 might mediate pain-induced secretions of DA. Collectively, these results suggest two potential therapeutic targets to reduce alcohol consumption that need further validation. They also suggest a novel mechanism for the sedative effects of alcohol at moderate and high doses.

Alcohol represents the third leading cause of preventable death in the United States. Yet, despite its prevalent role in impeding human health, there is much to understand about how it elicits its effects on the body and how the body and brain change when an individual becomes physiologically dependent upon alcohol. The work presented herein represents an effort to elucidate the acute and chronic effects of alcohol on the nervous system. We investigate two specific protein pathways and their role in alcohol’s effects on the body. The first begins with brain-derived neurotrophic factor (BDNF), which acts on TrkB, and ends with KCC2. We demonstrate that BDNF expression is increased in the VTA during withdrawal from chronic but not acute alcohol exposure and that this increase persists for at least seven days. Concomitantly, we demonstrate that the activation of GABAA channels on produces less inhibition of VTA GABA neurons in mice treated with chronic intermittent ethanol exposure than in alcohol naïve mice. This effect likewise persisted for at least seven days. We illustrate that BDNF has no apparent direct effect on VTA GABA neuron firing rate. The second pathway begins with the T cell marker CD5 and ends with the anti-inflammatory cytokine, IL-10. We demonstrate that in a genetic CD5 knockout (CD5 KO) mouse model both alcohol consumption as well as the sedative properties of alcohol are reduced. Since CD+ B cells secrete more IL-10 than CD5- B cells, we also demonstrate the effects of IL-10 on VTA neurons. We show that IL-10 has direct effects on VTA dopamine (DA) neurons by increasing their firing activity. We relatedly illustrate that IL-10 produces an increase in DA release in the nucleus accumbens (NAc). However, contrary to our hypotheses, we show that IL-10 produces conditioned place aversion rather than conditioned place preference in a place conditioning paradigm, suggesting that IL-10 might mediate pain-induced secretions of DA. Collectively, these results suggest two potential therapeutic targets to reduce alcohol consumption that need further validation. They also suggest a novel mechanism for the sedative effects of alcohol at moderate and high doses.