Obesity is associated with deficits in cognitive function, particularly within the domain of executive function (EF). EF refers to higher order cognitive processes that regulate our ability to sustain attention, inhibit subconscious tendencies, remember and manipulate information for immediate use, and remain cognitively flexible. Deficits in EF in overweight and obese individuals may impact the success of weight loss and maintenance efforts. Therefore, understanding the biological links between obesity and EF, as well as the ability to reverse EF deficits with weight loss, is imperative. The first study aimed to determine the effect of weight loss in overweight and obese, middle-aged and older adults on serum brain-derived neurotrophic fact (BDNF), S100 calcium binding protein B (S100B), and glial fibrillary acidic protein (GFAP). Serum samples (n=21; 50-75 years, BMI 25-40 kg/m2) were pooled from two prior weight loss studies. Fasting blood measurements were taken before and after 8- or 12-weeks of hypocaloric diet-induced weight loss (1200 or 1500 kcal/d). Body Mass Index (BMI), body weight, waist circumference, and percent body fat (All p<0.001) decreased with weight loss. Serum BDNF (p=0.871), S100B (p=0.898), and GFAP (p=0.506) did not change following weight loss. The second study aimed to determine the correlation between the magnitude of change in serum BDNF, S100B, and GFAP and the magnitude of improvement in EF performance on three computer-based tests. Participants (n=8; 50-75 years, BMI 25-40 kg/m2) completed 4-weeks of hypocaloric diet-induced weight loss (1200 or 1500 kcal/d), followed by 4-weeks of weight maintenance (hypocaloric diet + steps/d goal). Fasting blood and EF measurements were completed at baseline, and weeks 4 and 8. BMI (p=0.001), body weight (p=0.001), waist circumference (p=0.002), and percent body fat (p=0.001) decreased from baseline to week 8. Serum BDNF (p=0.359), S100B (p=0.277), and GFAP (p=0.585) did not change following weight loss. Go/No-Go (GNG) errors of commission (p=0.009) and AX-Continuous Performance Test (AX-CPT) correct response time (p=0.041) decreased following the weight loss. The change in serum GFAP was inversely correlated with GNG errors of omission (r=-0.716, p=0.046) and AX-CPT correct hits (r=-0.737, p=0.037), and positively correlated with AX-CPT correct response time (r=0.859, p=0.006). In conclusion, although weight loss does not influence serum BDNF, S100B, or GFAP levels, it may have a positive effect on inhibitory control in overweight and obese, middle-aged and older adults. Further research is needed to understand the relationship between serum BDNF, S100B, and GFAP and executive function. / Master of Science / Obesity is associated with lower brain function, particularly in executive function (EF). EF refers to advanced thought processes that help to maintain focus, practice self-control, solve problems, and easily switch between tasks. Lower EF in individuals with overweight and obesity may impact the success of weight loss and maintenance efforts. Because of this, understanding body processes that may link obesity and lower EF, as well as the ability to improve EF with weight loss, is very important. The first study aimed to determine the effect of weight loss on blood proteins responsible for brain health: brain-derived neurotrophic fact (BDNF), S100 calcium binding protein B (S100B), and glial fibrillary acidic protein (GFAP). Twenty-one blood samples from overweight and obese, middle-aged and older adults were combined from two completed weight loss studies. In these studies, blood was measured before and after 8- or 12-weeks of a weight loss (low calorie diet;1200 or 1500 Calories per day). Body Mass Index (BMI), body weight, waist circumference, and percent body fat all decreased with weight loss; however, levels of BDNF, S100B, and GFAP in the blood did not change. The second study aimed to determine the relationship between blood BDNF, S100B, and GFAP and performance on three computer-based tests of EF before and after weight loss. Eight overweight and obese, middle-aged and older adults completed 4-weeks of weight loss (low-calorie diet; 1200 or 1500 Calories per day), followed by 4-weeks of weight maintenance. BMI, body weight, waist circumference, and percent body fat all decreased following the weight loss and maintenance intervention (week 8). Blood BDNF, S100B, and GFAP levels did not change, but performance on two EF measures improved: participants made less errors of commission (doing something when not supposed to) and had faster reaction time following the intervention, indicating better self-control. Additionally, greater increases in GFAP were associated with less errors of omission (not doing something when supposed to), fewer correct responses, and slower reaction time. In conclusion, although weight loss did not affect blood BDNF, S100B, or GFAP levels, it may improve self-control in overweight and obese, middle-aged and older adults. Further research is needed to understand the relationship between weight loss, blood proteins of brain health, and EF.

Mild traumatic brain injury (mTBI), often referred to as concussion, has become increasingly recognized as a serious health issue in the general population. The prevalence of mTBI in athletes, particularly repeated injuries in young athletes, is of great concern as injuries to the developing brain can have long-term detrimental effects. In this study we used a novel awake closed-head injury (ACHI) model in rodents to examine repeated mTBI (rmTBI), to determine if repeated injuries produced the neurological and molecular changes evident with human concussion. Animals were administered 4, 8, and 16 rmTBIs and acute neurological assessments were performed after the injuries. Changes in glial fibrillary acidic protein (GFAP) and ionized calcium-binding adapter molecule 1 (Iba-1) levels were assessed using Western blot analysis at one day following rmTBI in the ipsilateral dentate gyrus (DG) and the cornu ammonis (CA) regions of the hippocampus and the cortex (CX) indicative of astrocyte and microglial cell reactivity. Results indicated that the ACHI model produces neurological deficits immediately after the injuries, with the most deficits arising in the rmTBI16 group. Despite deficits in all injury groups, histological staining with cresyl violet revealed no significant morphological tissue damage to the brain. Western blot analysis, however, showed a significant increase in DG and CX GFAP expression in the rmTBI16 group with no changes in Iba-1 levels. This suggests an acute activation of astrocytes in response to injury, with a delay or absence of microglial activation. Our findings show that with repetitive concussions, we are able to detect acute neurological and molecular changes in the juvenile female brain. However, further investigation is necessary to determine if these are transient changes. / Graduate

No description available.

No description available.

Objective: Activated astroglia is involved in the pathophysiology of neurodegenerative diseases and has also been described in animal models of spinal muscular atrophy (SMA). Given the urgent need of biomarkers for treatment monitoring of new RNA-modifying and gene replacement therapies in SMA, we examined glial fibrillary acidic protein concentrations in cerebrospinal fluid (cGFAP) as a marker of astrogliosis in SMA. - Methods: 58 adult patients and 21 children with genetically confirmed 5q-associated SMA from four German motor neuron disease specialist care centers and 30 age- and sex-matched controls were prospectively included in this study. cGFAP was measured and correlated to motor performance and disease severity. Additionally, we compared fL). - Results: cGFAP concentrations did not differ from controls but showed higher levels in more severely affected patients after adjustment for patients’ age. Normalized cNfL values were associated with disease severity. Within 14 months of nusinersen treatment, cGFAP concentrations did not change, while cNfL decreased significantly. - Interpretation: cGFAP is not an outstanding biomarker in SMA, but might support the hypothesis that glial activation is involved in SMA pathology. Unlike previously suggested, cNfL may be a promising biomarker also in adult patients with SMA, which should be subject to further investigations.

INTRODUÇÃO: As doenças retinianas associadas à neovascularização, tais como a degeneração macular relacionada à idade e as retinopatias diabética e da prematuridade são as principais e mais importantes causas da cegueira em todo o mundo. Nos últimos anos, injeções intravítreas de fármacos com ação antiangiogênica, como o bevacizumabe (BVZ), têm sido de grande valia tanto em pacientes na fase adulta quanto nos recém-natos. Todavia, estudos experimentais in vitro e in vivo sugerem que essas drogas promovam efeitos adversos sobre alguns processos celulares, interferindo diretamente em mecanismos fisiológicos que mantém a homeostase do tecido retiniano, incluindo os mecanismos de proliferação, diferenciação e morte celular. OBJETIVO: investigar o efeito do BVZ nos processos de transcrição e tradução de marcadores da gliose: GFAP e vimentina, de morte celular, caspase-3 e beclina-1, e dos proteoglicanos relacionados à manutenção e desenvolvimento de tecido retiniano: neurocam, fosfacam e sindecam-3. MÉTODOS: Dois modelos experimentais foram usados nesse estudo: 1) linhagem celular de Müller de Glia humana adulta (MIO-M1), cultivada em meio de cultura D-MEM na presença e ausência de BVZ por 12 e 24 horas nas concentrações de 0,25 mg/mL e 0,50 mg/mL e 2) explantes de retinas de ratos 2 dias pós-nascidos submetidos à 0,50 mg/mL da droga por 48 horas. Durante este período foram mantidos a 5% de dióxido de carbono à temperatura de 37°C. A análise de proteínas foi realizada por imunocitohistoquímica e Western Blotting e a expressão de RNAm, pela reação em cadeia da polimerase em tempo real (PCR Real Time). Foi utilizado o Teste de ANOVA - fator único para a comparação entre os grupos controle e tratados com BVZ de um mesmo período (12h ou 24h) e o teste t de Student para a comparação entre as mesmas concentrações de 12h e 24h, e para a comparação entre os grupos controle e tratado com BVZ dos explantes (p < 0,05). RESULTADOS: Nas células MIO-M1, o BVZ, aumentou a expressão gênica e diminui a tradução de VEGF na concentração de 0,50 mg/mL em 24h comparado a 12h. Para o GFAP, houve um aumento da transcrição em 0,50 mg/mL em 24h comparado a 12h e aos outros grupos em 24h. Entretanto, houve diminuição da tradução para estes mesmos períodos e condições. Para a vimentina, houve aumento na transcrição em 0,50 mg/mL após 24h. Os achados de beclina-1 revelaram uma diminuição da transcrição e tradução em 0,25 mg/mL em 24h comparado a 12h. A transcrição entre os grupos do mesmo período aumentou nos grupos tratados com BVZ tanto em 12h quanto em 24h. A tradução da beclina-1 diminuiu em 0,25 mg/mL, mas aumentou em 0,50 mg/mL em 24h em relação à 12h. A comparação entre os grupos de 24h revelou aumento da tradução em 0,50 mg/mL. Para a caspase-3, houve diminuição da transcrição em 0,25 mg/mL e 0,50 mg/mL em 24h em relação a 12h e entre nos grupos tratados com BVZ em 24h. A tradução revelou um aumento em 0,50 mg/mL em 24h em relação a 12h. No fosfacam, houve diminuição da transcrição em 0,50 mg/mL em 24h comparado a 12h e entre os grupos tratados com BVZ e controles para 12h e 24h. A transcrição de neurocam diminuiu em 0,25 mg/mL e 0,50 mg/mL em 24h comparado a 12h e entre os grupos tratados com BVZ e controles em 12h e 24h. A tradução aumentou em 0,50 mg/mL em 24h em relação a 12h, mas diminuiu entre os grupos em 24h. Nos explantes, a transcrição e tradução de VEGF diminuiram no grupo tratado com BVZ após 48h. CONCLUSÃO: Nossos resultados relacionados às células MIO-M1 e ao explante de ratos, in vitro, nos permitem aventar o possível comprometimento ocasionado pela depleção do VEGF pelo BVZ na homeostase do tecido retiniano, in vivo, interferindo nas moléculas envolvidas na morte e diferenciação celular e na neuroproteção em indivíduos em fase adulta e recém-nato / Backgraound: Vasoproliferative retinal disorders such as age-related macular, degeneration, diabetic retinopathy and retinopathy of prematurity are major causes of blindness in the world. In recent years, intravitreal injections of drugs with antiangiogenic action, as bevacizumab, have been very useful for both patients in adulthood and in newborns. However, experimental studies, in vivo and in vitro, suggest that antiangiogenic drugs may promote side effects in cellular proceedings, interfering directly in physiological mechanisms of cellular proliferation, differentiation and death. POURPOSE: Investigate the bevacizumab effects in transcription and translation processes of gliosis, GFAP and vimentin, cellular death markers, caspase-3 and beclin-1, and proteoglycans involved in retinal tissue maintenance and development, neurocan, phosphacan and syndecan-3. METHODS: Two experimental models were used on this research: cellular lineage of adult and human Müller glial cell(MIO-M1) were cultivated on D-MEM medium with 0,25 and 0,50 mg/mL bevacizumab for 12 and 24 hours, and two days old rat retinal explants submitted to 0,50 mg/mL for 48 hours. During this period were stored in laboratory ovens at 5% carbon dioxide pressure and 37 °C average temperature. Molecular techniques were used to evaluate gene expression and protein content. Protein assessments were performed by immunocytochemistry and western blotting analysis, while Real Time PCR was used to measure mRNA content. ANOVA tests one factor were applied to compare the control and BVZ groups of the same period (12h or 24h) and t test from Student to compare the same conditions of 12h and 24h, and to compare the control and BVZ retinal explants groups (p<0.05). RESULTS: At MIO-M1 cells, BVZ increased the gene expression and reduced the translation of VEGF at concentration of 0.50 mg / mL in 24 hours compared to 12 hours. For GFAP, there was an increase of transcription at 0.50 mg / mL in 24 hours compared to 12 hours and to the other groups at 24 hours. However, there was a decrease in translation for these same periods and conditions. For vimentin, there was an increase in transcription at 0.50 mg / mL after 24 hours. The beclin-1 findings revealed a decrease of transcription and translation at 0.25 mg / ml compared at 24 h compared to 12h. Transcription among groups increased in BVZ treated groups at 12h and 24h. The translation of beclin-1 decreased at 0.25 mg / ml, but increased at 0.50 mg / mL at 24 hours compared to 12 hours. The comparison between the groups at 24h revealed an increased in translation at 0.50 mg / mL. For caspase-3, there was a decrease in transcription at 0.25 mg / ml and 0.50 mg / ml at 24 compared to 12 hours and among BVZ treated groups at 24h. Translation revealed an increase at 0.50 mg / mL at 24 hours compared to 12 hours. For fosfacam, there was a decreased in transcription at 0.50 mg / mL in 24 hours compared to 12 hours and among BVZ treated groups and controls at 12h and 24h. The transcription of neurocam decreased at 0.25 mg / ml and 0.50 mg / ml at 24 hours compared to 12 hours and among BVZ treated groups and controls at 12h and 24h. Translation increased at 0.50 mg / mL at 24 compared to 12 hours, but decreased among the groups at 24 hours. For explants, transcription and translation of VEGF decreased in the BVZ group treated after 48h. CONCLUSION: Our results related to the MIO-M1 cells and explants of rats,in vitro, allow us to suggest the possible impairment caused by depletion of VEGF by BVZ in the homeostasis of retinal tissue, in vivo, interfering in the molecules involved in cell death and cell differentiation and neuroprotection in individuals in adulthood and newborns

INTRODUÇÃO: As doenças retinianas associadas à neovascularização, tais como a degeneração macular relacionada à idade e as retinopatias diabética e da prematuridade são as principais e mais importantes causas da cegueira em todo o mundo. Nos últimos anos, injeções intravítreas de fármacos com ação antiangiogênica, como o bevacizumabe (BVZ), têm sido de grande valia tanto em pacientes na fase adulta quanto nos recém-natos. Todavia, estudos experimentais in vitro e in vivo sugerem que essas drogas promovam efeitos adversos sobre alguns processos celulares, interferindo diretamente em mecanismos fisiológicos que mantém a homeostase do tecido retiniano, incluindo os mecanismos de proliferação, diferenciação e morte celular. OBJETIVO: investigar o efeito do BVZ nos processos de transcrição e tradução de marcadores da gliose: GFAP e vimentina, de morte celular, caspase-3 e beclina-1, e dos proteoglicanos relacionados à manutenção e desenvolvimento de tecido retiniano: neurocam, fosfacam e sindecam-3. MÉTODOS: Dois modelos experimentais foram usados nesse estudo: 1) linhagem celular de Müller de Glia humana adulta (MIO-M1), cultivada em meio de cultura D-MEM na presença e ausência de BVZ por 12 e 24 horas nas concentrações de 0,25 mg/mL e 0,50 mg/mL e 2) explantes de retinas de ratos 2 dias pós-nascidos submetidos à 0,50 mg/mL da droga por 48 horas. Durante este período foram mantidos a 5% de dióxido de carbono à temperatura de 37°C. A análise de proteínas foi realizada por imunocitohistoquímica e Western Blotting e a expressão de RNAm, pela reação em cadeia da polimerase em tempo real (PCR Real Time). Foi utilizado o Teste de ANOVA - fator único para a comparação entre os grupos controle e tratados com BVZ de um mesmo período (12h ou 24h) e o teste t de Student para a comparação entre as mesmas concentrações de 12h e 24h, e para a comparação entre os grupos controle e tratado com BVZ dos explantes (p < 0,05). RESULTADOS: Nas células MIO-M1, o BVZ, aumentou a expressão gênica e diminui a tradução de VEGF na concentração de 0,50 mg/mL em 24h comparado a 12h. Para o GFAP, houve um aumento da transcrição em 0,50 mg/mL em 24h comparado a 12h e aos outros grupos em 24h. Entretanto, houve diminuição da tradução para estes mesmos períodos e condições. Para a vimentina, houve aumento na transcrição em 0,50 mg/mL após 24h. Os achados de beclina-1 revelaram uma diminuição da transcrição e tradução em 0,25 mg/mL em 24h comparado a 12h. A transcrição entre os grupos do mesmo período aumentou nos grupos tratados com BVZ tanto em 12h quanto em 24h. A tradução da beclina-1 diminuiu em 0,25 mg/mL, mas aumentou em 0,50 mg/mL em 24h em relação à 12h. A comparação entre os grupos de 24h revelou aumento da tradução em 0,50 mg/mL. Para a caspase-3, houve diminuição da transcrição em 0,25 mg/mL e 0,50 mg/mL em 24h em relação a 12h e entre nos grupos tratados com BVZ em 24h. A tradução revelou um aumento em 0,50 mg/mL em 24h em relação a 12h. No fosfacam, houve diminuição da transcrição em 0,50 mg/mL em 24h comparado a 12h e entre os grupos tratados com BVZ e controles para 12h e 24h. A transcrição de neurocam diminuiu em 0,25 mg/mL e 0,50 mg/mL em 24h comparado a 12h e entre os grupos tratados com BVZ e controles em 12h e 24h. A tradução aumentou em 0,50 mg/mL em 24h em relação a 12h, mas diminuiu entre os grupos em 24h. Nos explantes, a transcrição e tradução de VEGF diminuiram no grupo tratado com BVZ após 48h. CONCLUSÃO: Nossos resultados relacionados às células MIO-M1 e ao explante de ratos, in vitro, nos permitem aventar o possível comprometimento ocasionado pela depleção do VEGF pelo BVZ na homeostase do tecido retiniano, in vivo, interferindo nas moléculas envolvidas na morte e diferenciação celular e na neuroproteção em indivíduos em fase adulta e recém-nato / Backgraound: Vasoproliferative retinal disorders such as age-related macular, degeneration, diabetic retinopathy and retinopathy of prematurity are major causes of blindness in the world. In recent years, intravitreal injections of drugs with antiangiogenic action, as bevacizumab, have been very useful for both patients in adulthood and in newborns. However, experimental studies, in vivo and in vitro, suggest that antiangiogenic drugs may promote side effects in cellular proceedings, interfering directly in physiological mechanisms of cellular proliferation, differentiation and death. POURPOSE: Investigate the bevacizumab effects in transcription and translation processes of gliosis, GFAP and vimentin, cellular death markers, caspase-3 and beclin-1, and proteoglycans involved in retinal tissue maintenance and development, neurocan, phosphacan and syndecan-3. METHODS: Two experimental models were used on this research: cellular lineage of adult and human Müller glial cell(MIO-M1) were cultivated on D-MEM medium with 0,25 and 0,50 mg/mL bevacizumab for 12 and 24 hours, and two days old rat retinal explants submitted to 0,50 mg/mL for 48 hours. During this period were stored in laboratory ovens at 5% carbon dioxide pressure and 37 °C average temperature. Molecular techniques were used to evaluate gene expression and protein content. Protein assessments were performed by immunocytochemistry and western blotting analysis, while Real Time PCR was used to measure mRNA content. ANOVA tests one factor were applied to compare the control and BVZ groups of the same period (12h or 24h) and t test from Student to compare the same conditions of 12h and 24h, and to compare the control and BVZ retinal explants groups (p<0.05). RESULTS: At MIO-M1 cells, BVZ increased the gene expression and reduced the translation of VEGF at concentration of 0.50 mg / mL in 24 hours compared to 12 hours. For GFAP, there was an increase of transcription at 0.50 mg / mL in 24 hours compared to 12 hours and to the other groups at 24 hours. However, there was a decrease in translation for these same periods and conditions. For vimentin, there was an increase in transcription at 0.50 mg / mL after 24 hours. The beclin-1 findings revealed a decrease of transcription and translation at 0.25 mg / ml compared at 24 h compared to 12h. Transcription among groups increased in BVZ treated groups at 12h and 24h. The translation of beclin-1 decreased at 0.25 mg / ml, but increased at 0.50 mg / mL at 24 hours compared to 12 hours. The comparison between the groups at 24h revealed an increased in translation at 0.50 mg / mL. For caspase-3, there was a decrease in transcription at 0.25 mg / ml and 0.50 mg / ml at 24 compared to 12 hours and among BVZ treated groups at 24h. Translation revealed an increase at 0.50 mg / mL at 24 hours compared to 12 hours. For fosfacam, there was a decreased in transcription at 0.50 mg / mL in 24 hours compared to 12 hours and among BVZ treated groups and controls at 12h and 24h. The transcription of neurocam decreased at 0.25 mg / ml and 0.50 mg / ml at 24 hours compared to 12 hours and among BVZ treated groups and controls at 12h and 24h. Translation increased at 0.50 mg / mL at 24 compared to 12 hours, but decreased among the groups at 24 hours. For explants, transcription and translation of VEGF decreased in the BVZ group treated after 48h. CONCLUSION: Our results related to the MIO-M1 cells and explants of rats,in vitro, allow us to suggest the possible impairment caused by depletion of VEGF by BVZ in the homeostasis of retinal tissue, in vivo, interfering in the molecules involved in cell death and cell differentiation and neuroprotection in individuals in adulthood and newborns