Background: The 18 kDa translocator protein (TSPO), expressed at a low level in the healthy human central nervous system (CNS), is upregulated in inflammatory brain diseases by activated microglia and other immune cells. Using positron emission tomography (PET) radioligands targeting TSPO, it is possible to localise this signal and map the course of microglial activation and its effects on disease progression. Here, a newly developed second generation TSPO PET ligand, [18F]GE-180, was evaluated in different models of preclinical and clinical neuroinflammatory disease. Methods: A preclinical model of low-level inflammation with lipopolysaccharide (LPS) was designed. Rats were scanned with the first generation TSPO ligand [11C]- (R)-PK11195 and either [18F]GE-180 or [18F]DPA-714, with dual scanning enabling the direct comparison of second generation tracers with [11C]-(R)-PK11195. An arterial blood sampling system for rodent imaging with [18F]DPA-714 was set up and characterised. The performance of [18F]GE-180 was assessed in a clinical study in nine relapsing-remitting multiple sclerosis patients (RRMS) and ten healthy volunteers (HV). A comparison of kinetic modelling approaches for [18F]GE-180 human brain PET data was performed, as well as a longitudinal analysis with intervention using the disease-modifying treatment, natalizumab to evaluate the potential of [18F]GE-180 as a biomarker for therapy monitoring in MS subjects. Finally, the plasma-protein binding behaviour of [18F]GE-180 was evaluated in vitro using ultrafiltration. Results: In LPS animals, [18F]GE-180 produced a significantly higher ipsi- to contralateral uptake ratio and binding potential () than [11C]-(R)-PK11195 (p = 0.03), but [18F]DPA-714 did not. There was no significant difference between animals scanned with [18F]GE-180 and [18F]DPA-714, suggesting no overall superiority of the former. Characterisation of an arterial sampling system for rodent studies with [18F]DPA-714 allowed correction for dispersion effects. A comparison of reference regions showed that a novel externally derived tissue estimated with lower bias than a contralateral reference region. In human [18F]GE-180 brain PET data, the unconstrained two-tissue compartment model (2TCM) best described tracer behaviour in RRMS and HV subjects. Normal appearing white matter (NAWM) in patients was elevated over that of HVs. Standardised uptake values (SUVs) for the tracer in rodents were 0.28±0.12 and 0.84±0.31 in healthy tissue and LPS lesions respectively, and in humans were 0.36±0.04 (HV) and 0.58 (in a gadolinium- enhancing MS lesion). [18F]GE-180 uptake was also significantly reduced in the brains of RRMS subjects treated with natalizumab, correlating with clinically-identified improvement. [18F]GE-180 has a free fraction of between 1 and 8%.Conclusions: [18F]GE-180 shows good brain uptake in the rodent brain and produces superior signal to [11C]-(R)-PK11195, but not to [18F]DPA-714. The 2TCM fits human [18F]GE-180 PET data well, and the tracer is able to identify an elevated signal in RRMS patients compared to healthy subjects. [18F]GE-180 shows a large fraction of non-displaceable binding in human blood, thus further optimisation of kinetic modelling approaches is suggested.

Text in Afrikaans / Hierdie studie handel oor die ontwerp van 'n post traumatiese terapie deur die opvoedkundige sielkundige waar troeteldiere as fasiliteerder ingespan kan word. Dit fokus op getraumatiseerdes wat na afloop van 'n trauma in 'n post traumatiese situasie verkeer, en nie klinies aan die diagnose van post traumatiese stres versteuring voldoen nie. Die ontwerp van die terapie val in vier fases uiteen wat nie noodwendig streng chronologies in terapie hoef te verloop nie. Eerstens is daar die affektiewe fase wat handel oor die bantering van emosies. Tweedens fokus die kognitiewe fase op kognitiewe herstrukturering by die getraumatiseerde en veral die internalisering daarvan. Die fase van voorraadopname maak voorsiening vir die identifisering van psigologiese gestremdhede en sterker modaliteite van die getraumatiseerde. Die kompensatoriese fase handel oor die ontwerp van 'n aksieplan vir die sinvolle voortsetting van die getraumatiseerde se lewe asook inoefeningsaspekte daarvan. Die terapeutiese ontwerp het beslag gekry na 'n uitgebreide literatuurstudie van verskeie terapeutiese skole se post traumatiese terapiee en tegnieke. Oorhoofs word bogenoemde ontwerp in die relasieterapie ingebed. Hierdie terapie le besondere klem op die kompensatoriese fase met die oog op adekwate aktualisering van die getraumatiseerde in sy toekomstige gesitueerdheid. Alhoewel daar 'n uitgebreide beskrywing van troeteldier gefasiliteerde terapie volg, het die navorser slegs sekere getraumatiseerde kliente ge'identifiseer wat sat baat by troeteldier gefasiliteerde terapie. Daar is tydens die studie aandag gegee aan die ontwerp van 'n vraelys wat as operasionele metingsinstrument dien om die omvang van die trauma ten opsigte van die getraumatiseerde se filnksionering, selfgesprekke, belewenis, betekenisgewing, betrokkenheid, seltkonsep, relasies en selfaktualisering te bepaal. Fasiliteerders van die affektiewe fase sluit onder andere soos reeds genoem hierbo, troeteldiere in. Ondersoek is ingestel na die riglyne vir troeteldier gefasiliteerde terapie, en verskeie aspekte soos determinante by die getraumatiseerde vir troeteldier gefasiliteerde terapie, die aard van mens-dier interaksie, risiko's en menslike voorwaardes verbonde aan troeteldier gefasiliteerde terapie, is beskryf Tydens die empiriese ondersoek is gevind dat alhoewel die ontwerp beperkinge het, daar besliste ruimte vir troeteldier gefasiliteerde post traumatiese terapie op Sielkundige Opvoedkunde terrein bestaan / This study deals with the design of a post traumatic therapy that can be administered by the educational psychologist. Pets were used for the first time as facilitators in such a therapy. This therapy was primarily designed for persons in a post traumatic situation after an experienced trauma, who _do not ~fy foc!h<L~!i_ni~_ctl _ cri!~!"ia gf post traumatic _stress disorder. Four phases can be distinguished in the course of the therapy. This doesn't however imply a hierargical order. First of all the affective phase deals with the emotional side of the traumatised person. Secondly cognitive restructuring is taken up in the cognitive phase as well as internalization thereof The third phase of stock taking makes provision for the identification of psychological handicaps and stronger modalities of the traumatised person. A plan of action is developed in the final compensatory phase and it deals with optimal future actualization of the traumatised person. If necessary, opportunity is provided for excercising the newly required skills. This design was developed after extensive literature studies of post traumatic therapies and techniques, and it is embedded in the overall approach of relationship therapy. Strong emphasis is placed on the compensatory phase in view of the traumatised person's adequate future actualization. Although an extensive description of pet fasilitated therapy is given, only certain traumatised clients were identified who will benefit from this therapy. A questionnaire as operational measuring instrument was developed for the purpose of this study. Information as to the traumatised person's functioning, selftalk, experiences, allocation of meaning, involvement, selfconcept, relationship formation and selfactualization can be ascertained via this instrument. One of the facilitators of the affective phase, already mentioned, includes pets. A thorough study into guidelines for pet facilitated therapy, as well as aspects such as determinants of traumatised people as indication for this kind of therapy, the nature of human-animal interaction, risks and human conditions for pet facilitated therapy, was undertaken. The empirical study proved that in spite of limitations of pet facilitated post traumatic therapy, definite opportunity exists in the field of Psychology of Education for this kind of therapy / D.Ed. (Sielkundige Opvoedkunde)

O uso de tetracloreto de carbono (CCl4) em ratos é um modelo experimental de dano ao tecido hepático, desencadeando fibrose e, a longo prazo, cirrose. A cirrose hepática é uma doença crônica progressiva que representa um estado de disfunção hepática irreversível ou lentamente reversível, caracterizado pela formação de nódulos fibróticos. Este estudo possui como objetivo avaliar as alterações hepáticas e pulmonares causadas pelo modelo de cirrose hepática através da utilização de CCl4 intraperitoneal. Foram utilizados 18 ratos Wistar machos divididos em 3 grupos: grupo controle (CO) e outros 2 grupos divididos pelo tempo de indução da cirrose por CCl4. G1 (11 semanas), G2 (16 semanas). Verificamos a elevação significativa no nível das transaminases hepáticas, na lipoperoxidação do tecido hepático e pulmonar (TBARS) e nas enzimas antioxidantes SOD e CAT, além de um aumento da expressão de TNF-M e IL-1N no pulmão dos animais cirróticos. Observamos alteração nas trocas gasosas de ambos os grupos cirróticos. Podemos concluir que nosso modelo reproduziu a cirrose hepática, além de causar alterações no sistema pulmonar, provocando alteração nas trocas gasosas e alterando o tamanho dos vasos pulmonares. / The use of carbon tetrachloride (CCl4) in rats is an experimental model of hepatic tissue damage, which leads to fibrosis, and at the long term, cirrhosis. Cirrhosis is a chronic progressive disease that represents a state of irreversible or slowly reversible hepatic disfunction, characterized by fibrotic nodules formation. The aim of this study is to evaluate the changes caused by cirrhosis in lung and liver, through the experimental model of intraperitoneal CCl4 administration. We used 18 male Wistar rats divided into three groups: control (CO) and two groups divided by the time of cirrhosis induction by CCl4. G1 (11semanas), G2 (16semanas). We found significant increase of transaminase levels and lipid peroxidation (TBARS) in liver and lung tissue. Also, increased antioxidant enzymes SOD and CAT, as well as the expression of TNF-M and IL-1N in the lung of cirrhotic animals. We observed changes in gas exchange in both cirrhotic groups. We can conclude that our model reproduces the liver cirrhosis, that causes alterations in the pulmonary system that leads to changes in gas exchange and size of pulmonary vessels.

CoordenaÃÃo de AperfeiÃoamento de NÃvel Superior / A depressÃo à uma doenÃa recorrente e incapacitante cujo tratamento clÃnico està relacionado com modulaÃÃes nos sistemas monoaminÃrgicos em diversas Ãreas cerebrais. A riparina II (ripII), alcamida isolada do fruto verde de Aniba riparia, apresentou previamente efeito antidepressivo em modelos comportamentais. Dessa forma, objetivando investigar o potencial antidepressivo da ripII, foram realizados experimentos comportamentais como o teste do nado forÃado, suspensÃo da cauda e campo aberto. Para avaliar o envolvimento do sistema monoaminÃrgico, os animais foram prÃ-tratados com antagonistas especÃficos para receptores 5-HT1A, 5-HT2A/2C e 5-HT3 de serotonina (5-HT), D1 e D2 de dopamina (DA) e a1 e a2 de noradrenalina (NA) no teste do nado forÃado. AlÃm disso, os animais previamente tratados com ripII e submetidos ao teste do nado forÃado tiveram as Ãreas cerebrais hipocampo, corpo estriado e cÃrtex prÃ-frontal retiradas para detecÃÃo dos nÃveis de monoaminas em HPLC eletroquÃmico ou para realizaÃÃo dos experimentos de estresse oxidativo, para o qual foram investigadas a atividade enzimÃtica da superÃxido dismutase, quantificaÃÃo dos nÃveis de glutationa reduzida (GSH) e nitrito/nitrato, alÃm do grau de lipoperoxidaÃÃo. A ripII foi administrada agudamente, por via oral, na dose de 50 mg/kg, em todos os testes realizados. Os resultados mostraram que a ripII apresentou efeito antidepressivo nos modelos de nado forÃado e suspensÃo da cauda sugerindo que este efeito seja especÃfico, uma vez que os animais nÃo apresentaram alteraÃÃes na atividade locomotora no teste do campo aberto. Na avaliaÃÃo dos sistemas monoaminÃrgicos, os resultados mostraram que os antagonistas SCH23390 (D1), sulpirida (D2), prazosina (a1), NAN-190 (5-HT1A) e ondansentron (5-HT3) reverteram o tempo de imobilidade da ripII no nado forÃado sugerindo a participaÃÃo desses receptores para o efeito antidepressivo da substÃncia, enquanto nÃo houve alteraÃÃo deste efeito na presenÃa dos antagonistas ioimbina (a2) e ritanserina (5-HT2A/2C) sugerindo a nÃo participaÃÃo desses receptores no efeito da droga. A administraÃÃo prÃvia de ripII, antes do nado forÃado, reverteu o aumento nos nÃveis de peroxidaÃÃo lipÃdica, mas aumentou os nÃveis de GSH em hipocampo, corpo estriado e cÃrtex prÃ-frontal. Em conclusÃo, o estudo sugere uma aÃÃo moduladora, exercida por ripII, sobre o funcionamento dos sistemas noradrenÃrgico, dopaminÃrgico e serotonÃrgico, em nÃvel central, como mecanismo para o efeito antidepressivo no teste do nado forÃado, bem como a participaÃÃo de propriedades antioxidantes diretas ou indiretas dessa droga, atravÃs da capacidade de modificar a resposta ao estresse oxidativo neuronal. / Depression is a disabling and recurrent disease whose clinical treatment is related to modulations in monoaminergic systems in various brain areas. Riparina II (ripII), alkamide isolated from unripe fruit of Aniba riparia, has shown previously antidepressant-like effects in animal behavioral models. Thus, in order to investigate the potential antidepressant ripII, behavioral experiments were performed as the forced swim, tail suspension and open field tests. To assess the involvement of monoaminergic system, animals were pretreated with specific antagonists to 5-HT1A-, 5-HT2A/2C-, and 5-HT3-serotonin (5-HT) receptors, to D1- and D2-dopamine (DA) receptors and to &#61537;1- and &#61537;2-noradrenaline (NA) receptors in the forced swimming test. Furthermore, animals pretreated with ripII and submitted to the forced swim test had their brain areas such as hippocampus, striatum and prefrontal cortex removed for detection of monoamine levels in HPLC electrochemical or to carry out the experiments of oxidative stress. In the oxidative stress assays we investigated enzymatic activities of superoxide dismutase, measured the levels of reduced glutathione (GSH) and nitrite/nitrate, and lipid peroxidation degree. RipII was acutely administered orally at a dose of 50 mg/kg in all tests. The results showed that ripII presented antidepressant effect on the forced swim and tail suspension tests suggesting that this effect is specific, since the animals showed no changes in locomotor activity in open field test. In the evaluation of monoaminergic systems, the results showed that the antagonists SCH23390 (D1), sulpiride (D2), prazosin (&#61537;1), NAN-190 (5-HT1A) and ondansentron (5-HT3) reversed the immobility time of ripII on the forced swim test suggesting the involvement of these receptors for the antidepressant effect of ripII, while no change of this effect in the presence of the antagonists yohimbine (&#61537;2) and ritanserin (5-HT2A/2C) was observed, suggesting non-participation of these receptors on the drug effect. The prior administration of ripII before the forced swimming, reversed the increased levels of lipid peroxidation and increased levels of GSH in hippocampus, striatum and prefrontal cortex. In conclusion, the study suggests a modulating action exerted by ripII on the functioning of the noradrenergic, dopaminergic and serotonergic levels in the brain, as a mechanism for the antidepressant effect in the forced swimming test, as well as the participation of direct or indirect antioxidant properties of this drug through the ability to modify the neuronal response to oxidative stress.

O uso de tetracloreto de carbono (CCl4) em ratos é um modelo experimental de dano ao tecido hepático, desencadeando fibrose e, a longo prazo, cirrose. A cirrose hepática é uma doença crônica progressiva que representa um estado de disfunção hepática irreversível ou lentamente reversível, caracterizado pela formação de nódulos fibróticos. Este estudo possui como objetivo avaliar as alterações hepáticas e pulmonares causadas pelo modelo de cirrose hepática através da utilização de CCl4 intraperitoneal. Foram utilizados 18 ratos Wistar machos divididos em 3 grupos: grupo controle (CO) e outros 2 grupos divididos pelo tempo de indução da cirrose por CCl4. G1 (11 semanas), G2 (16 semanas). Verificamos a elevação significativa no nível das transaminases hepáticas, na lipoperoxidação do tecido hepático e pulmonar (TBARS) e nas enzimas antioxidantes SOD e CAT, além de um aumento da expressão de TNF-M e IL-1N no pulmão dos animais cirróticos. Observamos alteração nas trocas gasosas de ambos os grupos cirróticos. Podemos concluir que nosso modelo reproduziu a cirrose hepática, além de causar alterações no sistema pulmonar, provocando alteração nas trocas gasosas e alterando o tamanho dos vasos pulmonares. / The use of carbon tetrachloride (CCl4) in rats is an experimental model of hepatic tissue damage, which leads to fibrosis, and at the long term, cirrhosis. Cirrhosis is a chronic progressive disease that represents a state of irreversible or slowly reversible hepatic disfunction, characterized by fibrotic nodules formation. The aim of this study is to evaluate the changes caused by cirrhosis in lung and liver, through the experimental model of intraperitoneal CCl4 administration. We used 18 male Wistar rats divided into three groups: control (CO) and two groups divided by the time of cirrhosis induction by CCl4. G1 (11semanas), G2 (16semanas). We found significant increase of transaminase levels and lipid peroxidation (TBARS) in liver and lung tissue. Also, increased antioxidant enzymes SOD and CAT, as well as the expression of TNF-M and IL-1N in the lung of cirrhotic animals. We observed changes in gas exchange in both cirrhotic groups. We can conclude that our model reproduces the liver cirrhosis, that causes alterations in the pulmonary system that leads to changes in gas exchange and size of pulmonary vessels.

No description available.

No description available.

No description available.

The global number of obese people has reached pandemic proportions. High caloric diets and reduced physical exercise are to blame for this growing epidemic. Obesity has a very complex association with several other metabolic disorders, such as insulin resistance (IR), diabetes mellitus type 2 (DMT2) and cardiovascular disease. This puts a huge burden on health care systems world wide and claims many lives. Sutherlandia frutescens is a traditionally used herb, which is known to have anti-diabetic properties. However, the direct mode of action of S. frutescens still remains to be elucidated. The aim of this study was to investigate the developmental stages of high fat diet (HFD)-induced IR, to illuminate the pathogenesis of IR with a focal point on modifications in the lipid metabolism. Furthermore, the effects of S. frutescens as a treatment or prevention drug for IR and associated metabolic changes were examined. Two sets of experiments were conducted on male Wistar rats. In the first experiment rats, one week post weaning received a low fat diet (LFD), high fat diet (HFD) or HFD supplemented with S. frutescens (50mg/kg BW/d). Rats were sacrificed at week 0, 1, 2, 4, 8 and 12 in the feeding regime. In a second experiment rats were fed with a LFD or a HFD for 12 weeks and treated thereafter with S. frutescens (50mg/kg BW/d), metformin (13mg/kg BW/d) or water (control) for 28 days. Rats in the second experiment were sacrificed at week 12 to confirm IR while concurrently run rats were sacrificed after 28 days of treatment. For all the experiments rats were anaesthetized, blood was removed and rats were dissected. Plasma samples were analyzed for insulin, glucose, blood lipid parameters and cytokines. Liver, muscle and adipose tissue were analyzed for glucose uptake, total lipid content, lipid profile and fatty acid profile. It was shown that the intake of HFD caused IR and hyperinsulinaemia. The developmental stages in experiment one confirmed that an increase in plasma free fatty acids preceeded the onset of IR. Plasma and tissue lipid parameters (free fatty acid-, triglyceride- and cholesterol concentrations) showed pathological modifications in the HFD group. An ectopic accumulation of fat was observed in muscle and liver, as well as a change in membrane fatty acid profile. The results for circulating cytokines were somewhat inconclusive. Rats supplemented with S. frutescens did not develop HFD-induced IR (study one) or IR was reversed (study two). S. frutescens treatment also resulted in positive changes in plasma and tissue lipid parameters. In summary, an animal model for HFD-induced IR was established and the detrimental effect of elevated plasma FFA on glucose and lipid metabolism was observed. A novel discovery suggests that the anti-diabetic mode of action of S. frutescens is through modulation of lipid metabolism. It was also established that S. frutescens has the potential to prevent IR in vivo.

Obesity can lead to Type 2 Diabetes, both conditions increase in association with physical inactivity and high-energy diets, resulting in elevated blood glucose, decreased insulin sensitivity and increased insulin resistance. Sutherlandia frutescens (S.frutescens), an anti-diabetic plant, reverses and prevents insulin resistance in a rat model and human cell culture model. Gene expression analysis in hepatocyte cultures, identified genes down regulated in insulin resistance and up regulated by S.frutescens. These included genes encoding vesicle transporter proteins, hypothesised to be linked to hepatic lipid accumulation and lipid droplet formation during insulin resistance. The aim of this study was to investigate critical genes involved in lipid droplet formation, vesicle assembly and transport in high fat diet (HFD)-induced insulin resistant rat liver tissue during the development of insulin resistance and the reversal of these changes by S.frutescens. Rats were fed a low fat diet (LFD) or HFD supplemented with S.frutescens for 2, 4 and 8 weeks. Rats fed a HFD for 12 weeks developed insulin resistance, confirmed by plasma glucose and insulin levels (compared to normal controls). Groups of these rats were gavaged with S. frutescens (50mg/kg BW), Metformin (13mg/kg BW) or water for a further 4 weeks and starved for 12 hours, anaesthetized and blood removed by heart puncture. Liver was stored in RNA-Later™ for qRT-PCR and snap-frozen in liquid nitrogen for western blotting and confocal microscopy analysis. Changes in expression of vesicle transporter genes VAMP3 and NSF were analysed by qRT-PCR and changes in the protein expression by western blotting analysis. Proteins were localised within the liver by confocal immunohistochemistry using ZEN lite™ software. Statistical analysis was performed using One-Way ANOVA and unpaired t-test. mRNA gene expression of vesicle transport components VAMP3, NSF and SNAP25 showed relatively moderate changes with considerable individual variation within control or experimental groups. Uncorrelated changes in mRNA and protein products were found and may be due to differential regulation by siRNA. Proteins also showed altered staining patterns in high fat diet rats that reverted towards normal on S. frutescens treatment, potentially reflecting functional changes associated with transport of lipid-filled vesicles.