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We have shown that activin A (activin), a TGF-ß superfamily member, has pro-metastatic effects in colorectal cancer (CRC). In lung cancer, activin activates pro-metastatic pathways to enhance tumor cell survival and migration while augmenting CD4+ to CD8+ communications to promote cytotoxicity. Here, we hypothesized that activin exerts cell-specific effects in the tumor microenvironment (TME) of CRC to promote anti-tumoral activity of immune cells and the pro-metastatic behavior of tumor cells in a cell-specific and context-dependent manner. We generated an Smad4 epithelial cell specific knockout (Smad4-/-) which was crossed with TS4-Cre mice to identify SMAD-specific changes in CRC. We also performed IHC and digital spatial profiling (DSP) of tissue microarrays (TMAs) obtained from 1055 stage II and III CRC patients in the QUASAR 2 clinical trial. We transfected the CRC cells to reduce their activin production and injected them into mice with intermittent tumor measurements to determine how cancer-derived activin alters tumor growth in vivo. In vivo, Smad4-/- mice displayed elevated colonic activin and pAKT expression and increased mortality. IHC analysis of the TMA samples revealed increased activin was required for TGF-ß-associated improved outcomes in CRC. DSP analysis identified that activin co-localization in the stroma was coupled with increases in T-cell exhaustion markers, activation markers of antigen presenting cells (APCs), and effectors of the PI3K/AKT pathway. Activin-stimulated PI3K-dependent CRC transwell migration, and the in vivo loss of activin lead to smaller CRC tumors. Taken together, activin is a targetable, highly context-dependent molecule with effects on CRC growth, migration, and TME immune plasticity.
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INTRODUCTION: Diet and decreased gut microbiome diversity has been associated with acute pancreatitis (AP) risk. However, differences in dietary intake, gut microbiome, and their impact on microbial end metabolites have not been studied in AP. We aimed to determine differences in (i) dietary intake (ii) gut microbiome diversity and sulfidogenic bacterial abundance, and (iii) serum short-chain fatty acid (SCFA) and hydrogen sulfide (H 2 S) concentrations in AP and control subjects. METHODS: This case-control study recruited 54 AP and 46 control subjects during hospitalization. Clinical and diet data and stool and blood samples were collected. 16S rDNA sequencing was used to determine gut microbiome alpha diversity and composition. Serum SCFA and H 2 S levels were measured. Machine learning (ML) model was used to identify microbial targets associated with AP. RESULTS: AP patients had a decreased intake of vitamin D 3 , whole grains, fish, and beneficial eicosapentaenoic, docosapentaenoic, and docosahexaenoic acids. AP patients also had lower gut microbiome diversity ( P = 0.021) and a higher abundance of sulfidogenic bacteria including Veillonella sp. and Haemophilus sp., which were associated with AP risk. Serum acetate and H 2 S concentrations were significantly higher in the AP group ( P < 0.001 and P = 0.043, respectively). ML model had 96% predictive ability to distinguish AP patients from controls. DISCUSSION: AP patients have decreased beneficial nutrient intake and gut microbiome diversity. An increased abundance of H 2 S-producing genera in the AP and SCFA-producing genera in the control group and predictive ability of ML model to distinguish AP patients indicates that diet, gut microbiota, and their end metabolites play a key role in AP.
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Microbioma Gastrointestinal , Pancreatite , Animais , Humanos , Pancreatite/etiologia , Estudos de Casos e Controles , Doença Aguda , Dieta , Ácidos Graxos VoláteisRESUMO
Those with mild cognitive impairment (MCI), a precursor to dementia, have a gut microbiome distinct from healthy individuals, but this has only been shown in healthy individuals, not in those exhibiting several risk factors for dementia. Using amplicon 16S rRNA gene sequencing in a case-control study of 60 older (ages 55-76), obese, predominately female, African American adults, those with MCI (cases) had different gut microbiota profiles than controls. While microbial community diversity was similar between cases and controls, the abundances of specific microbial taxa weren't, such as Parabacteroides distasonis (lower in cases) and Dialister invisus (higher in cases). These differences disappeared after adjusting for markers of oxidative stress and systemic inflammation. Cognitive scores were positively correlated with levels of Akkermansia muciniphila, a bacterium associated with reduced inflammation. Our study shows that gut microbial composition may be associated with inflammation, oxidative stress, and MCI in those at high risk for dementia.
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Disfunção Cognitiva , Microbioma Gastrointestinal , Obesidade , Idoso , Feminino , Humanos , Pessoa de Meia-Idade , Negro ou Afro-Americano , Estudos de Casos e Controles , Disfunção Cognitiva/microbiologia , Demência , Microbioma Gastrointestinal/genética , Inflamação , Obesidade/microbiologia , RNA Ribossômico 16S/genética , MasculinoRESUMO
Obesity is considered an independent risk factor for colorectal cancer (CRC). Altered nutrient metabolism, particularly changes to digestion and intestinal absorption, may play an important role in the development of CRC. Iron can promote the formation of tissue-damaging and immune-modulating reactive oxygen species. We conducted a crossover, controlled feeding study to examine the effect of three, 3-week diets varying in iron and saturated fat content on the colonic milieu and systemic markers among older females with obesity. Anthropometrics, fasting venous blood and stool were collected before and after each diet. There was a minimum 3-week washout period between diets. Eighteen participants consumed the three diets (72% Black; mean age 60.4 years; mean body mass index 35.7 kg/m2). Results showed no effect of the diets on intestinal inflammation (fecal calprotectin) or circulating iron, inflammation, and metabolic markers. Pairwise comparisons revealed less community diversity between samples (beta diversity, calculated from 16S rRNA amplicon sequences) among participants when consuming a diet low in iron and high in saturated fat vs. when consuming a diet high in iron and saturated fat. More studies are needed to investigate if dietary iron represents a salient target for CRC prevention among individuals with obesity.
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Dieta , Microbioma Gastrointestinal , Intestinos , Feminino , Humanos , Pessoa de Meia-Idade , Dieta Hiperlipídica/efeitos adversos , Ácidos Graxos , Inflamação/etiologia , Ferro , Obesidade/complicações , Obesidade/epidemiologia , Obesidade/microbiologia , RNA Ribossômico 16S/genética , Intestinos/microbiologia , Intestinos/fisiologiaRESUMO
Resumen: Introducción: estudios previos han mostrado una mayor incidencia de todas las causas de hemorragia postparto durante la pandemia de COVID-19. El acretismo placentario es una causa frecuente de hemorragia postparto. Objetivo: determinar las diferencias en la severidad del choque hemorrágico y la cantidad de sangrado en mujeres con diagnóstico del espectro placenta previa-acretismo, antes de la pandemia y durante la pandemia por COVID-19. Material y métodos: en un estudio con cohortes comparativas de mujeres con placenta previa-acretismo, atendidas en el Instituto Nacional de Perinatología, durante el periodo de enero de 2017 a diciembre de 2019, grupo I (prepandemia), y de enero de 2020 a marzo de 2022, grupo II (pandemia), se comparó entre los grupos la cantidad de hemorragia y la clase del choque hemorrágico de acuerdo a ATLS. Resultados: se estudió a 277 mujeres, 211 fueron del grupo I (pre-COVID-19) y 66 del grupo II (COVID-19), se observó una mayor cantidad de sangrado durante la pandemia de COVID versus la fase pre-COVID (2,150.56 ± 1,910.08 mL versus 1,246.34 ± 1,494.1 mL) p = 0.001, existieron diferencias en las proporciones de la gravedad del choque hemorrágico en la fase de pandemia para las clases III y IV. Conclusión: se encontró un incremento en la cantidad de sangrado y gravedad del choque durante la pandemia de COVID-19.
Abstract: Introduction: previous studies have shown an increased incidence of all-cause postpartum hemorrhage during the COVID-19 pandemic. Placental accreta is a frequent cause of hemorrhagic shock. Objective: determine the severity of shock and the amount of bleeding during the COVID-19 pandemic in women diagnosed with placenta previa and placental accreta. Material and methods: in a comparative cohort study of women with placenta previa and accreta treated at the National Institute of Perinatology during the period from January 2017 to December 2019 group I (pre-pandemic) and from January 2020 to March 2022 group II (pandemic) the amount of hemorrhage and the class of hemorrhagic shock according to ATLS were compared between the groups. Results: 277 women were studied, 211 were from group I (pre COVID-19) and 66 from group II (COVID-19). A greater amount of bleeding was observed during the COVID pandemic versus the pre COVID phase (2,150.56 ± 1,910.08 mL vs 1,246.34 ± 1,494.1 mL) p = 0.001, there were differences in the proportions of severity of hemorrhagic shock in the pandemic phase for classes III and IV. Conclusion: an increase in the amount of bleeding and severity of shock was found during the COVID-19 pandemic.
Resumo: Introdução: estudos anteriores mostraram uma maior incidência de hemorragia pós-parto de todas as causas durante a pandemia de COVID-19. O acretismo placentário é uma causa frequente de hemorragia pós-parto. Objetivo: determinar as diferenças na gravidade do choque hemorrágico e na quantidade de sangramento em mulheres diagnosticadas com o espectro de placenta prévia-acretismo antes da pandemia e durante a pandemia de COVID-19. Material e métodos: em um estudo de coorte comparativo de mulheres com placenta prévia e acreta tratadas no Instituto Nacional de Perinatologia durante o período de janeiro de 2017 a dezembro de 2019, grupo I (pré-pandemia) e de janeiro de 2020 a março de 2022, grupo I II (pandemia) a quantidade de sangramento e a classe de choque hemorrágico de acordo com o ATLS foram comparadas entre os grupos. Resultados: foram estudadas 277 mulheres, 211 eram do grupo I (pré-COVID-19) e 66 do grupo II (COVID-19). Observou-se uma maior quantidade de sangramento durante a pandemia de COVID versus a fase pré-COVID (2,150.56 ± 1,910.08 mL vs 1,246.34 ± 1,494.1 ml) p = 0.001, houve diferenças nas proporções da gravidade do choque hemorrágico na fase pandêmica para as classes III e IV. Conclusão: um aumento na quantidade de sangramento e gravidade do choque foi encontrado durante a pandemia de COVID-19.
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Peripheral inflammation has been implicated in cognitive dysfunction and dementia. While studies outline the relationship between elevated inflammation and individual gray or white matter alterations, less work has examined inflammation as related to connectivity between gray and white matter or variability in these associations by race. We examined the relationship between peripheral inflammation and tract-based structural connectomics in 74 non-demented participants (age = 69.19 ± 6.80 years; 53% female; 45% Black) who underwent fasting venipuncture and MRI. Serum was assayed for C-reactive protein, interleukin-6, and interleukin-1ß. Graph theory analysis integrated T1-derived gray matter volumes and DTI-derived white matter tractography into connectivity matrices analyzed for local measures of nodal strength and efficiency in a priori regions of interest associated with cardiovascular disease risk factors and dementia. Linear regressions adjusting for relevant covariates showed associations between inflammatory markers and nodal strength in the isthmus, posterior and caudal anterior cingulate (p's ≤ .042). Adding an inflammatory marker*race term showed race-modified associations between C-reactive protein and efficiency in the thalamus and amygdala, and nodal strength in the putamen (p's ≤ .048), between interleukin-6 and efficiency in the pars triangularis and amygdala (p's ≤ .024), and between interleukin-1ß and nodal strength in the pars opercularis (p = .048). Higher levels of inflammation associated with lower efficiency and higher strength for White participants but higher efficiency and lower strength for Black participants. Results suggest inflammation is associated with tract-based structural connectomics in an older diverse cohort and that differential relationships may exist by race within prefrontal and limbic brain regions.
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Conectoma , Substância Branca , Idoso , Encéfalo/diagnóstico por imagem , Feminino , Substância Cinzenta/diagnóstico por imagem , Humanos , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Substância Branca/diagnóstico por imagemRESUMO
BACKGROUND: Iron is critical for fetal development. Neonates of obese women may be at risk for poor iron status at birth as a result of maternal inflammation-driven overexpression of hepcidin. OBJECTIVES: The objective of this study was to determine differences in placental transfer of oral iron (57Fe) and expression of placental transferrin receptor 1 (TFR1) and ferroportin (FPN) mRNA and protein and their association with maternal and neonatal iron-related parameters, including maternal hepcidin, among women with and without prepregnancy (PP) obesity. METHODS: 57Fe ingested during the third trimester of pregnancy was recovered in venous umbilical cord blood among 20 PP obese [BMI (in kg/m2): 30.5-43.9] and 22 nonobese (BMI: 18.5-29.0) women aged 17-39 y. Placental TFR1 and FPN mRNA and protein expression were quantified via qPCR and Western blot. Maternal and neonatal markers of iron status and regulation, as well as inflammation, were measured. Descriptive and inferential statistical tests (e.g., Student t test, Pearson correlation) were used for data analysis. RESULTS: There was no difference in cord blood enrichment of 57Fe or placental mRNA or protein expression of TFR1 or FPN among the women with and without PP obesity. Maternal hepcidin was not correlated with cord blood enrichment of 57Fe or placental FPN mRNA or protein expression. Maternal log ferritin (corrected for inflammation) was inversely correlated with log percent enrichment of 57Fe in cord blood (partial r = -0.50; P < 0.01, controlled for marital status) and protein expression of TFR1 (r = -0.43; P = 0.01). CONCLUSIONS: Placental iron trafficking did not differ among women with and without PP obesity. Findings reinforce the importance of maternal iron stores in regulating placental iron trafficking.
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Ferro , Placenta , Feminino , Ferritinas , Sangue Fetal/metabolismo , Hepcidinas/genética , Hepcidinas/metabolismo , Humanos , Recém-Nascido , Ferro/metabolismo , Obesidade , Placenta/metabolismo , Gravidez , Terceiro Trimestre da GravidezRESUMO
Pancreatic ductal adenocarcinoma (PDAC) has extensive stromal involvement and remains one of the cancers with the highest mortality rates. Activin A has been implicated in colon cancer and its stroma but its role in the stroma of PDAC has not been elucidated. Activin A expression in cancer and stroma was assessed in human PDAC tissue microarrays (TMA). Activin A expression in human TMA is significantly higher in cancer samples, with expression in stroma correlated with shorter survival. Cultured pancreatic stellate cells (PSC) were found to secrete high levels of activin A resulting in PDAC cell migration that is abolished by anti-activin A neutralizing antibody. KPC mice treated with anti-activin A neutralizing antibody were evaluated for tumors, lesions and metastases quantified by immunohistochemistry. KPC mice with increased tumor burden express high plasma activin A. Treating KPC mice with an activin A neutralizing antibody does not reduce primary tumor size but decreases tumor metastases. From these data we conclude that PDAC patients with high activin A expression in stroma have a worse prognosis. PSCs secrete activin A, promoting increased PDAC migration. Inhibition of activin A in mice decreased metastases. Hence, stroma-rich PDAC patients might benefit from activin A inhibition.
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Ativinas/metabolismo , Neoplasias Pancreáticas/metabolismo , Neoplasias Pancreáticas/patologia , Ativinas/sangue , Adenocarcinoma/sangue , Adenocarcinoma/genética , Adenocarcinoma/patologia , Animais , Carcinoma Ductal Pancreático/sangue , Carcinoma Ductal Pancreático/genética , Carcinoma Ductal Pancreático/patologia , Linhagem Celular Tumoral , Movimento Celular , Modelos Animais de Doenças , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Epitélio/metabolismo , Regulação Neoplásica da Expressão Gênica , Humanos , Camundongos , Metástase Neoplásica , Neoplasias Pancreáticas/sangue , Neoplasias Pancreáticas/genética , Células Estreladas do Pâncreas/metabolismo , Células Estreladas do Pâncreas/patologia , Prognóstico , Células Estromais/metabolismo , Análise de Sobrevida , Carga Tumoral , Regulação para Cima/genéticaRESUMO
OBJECTIVES: Non-Latino Black adults have greater risk for Alzheimer's dementia compared to non-Latino White adults, possibly due to factors disproportionally affecting Black adults including cardiovascular disease (CVD). Chronic peripheral inflammation is implicated in both Alzheimer's dementia and CVD and is known to impact cognition and cerebral white matter, yet little work has examined these associations by race. This study examined associations between inflammation, cognition, and cerebral white matter generally, and by race. METHODS: Eighty-six non-demented older Black and White participants (age = 69.03; 50% female; 45% Black participants) underwent fasting venipuncture, cognitive testing, and MRI. Serum was assayed for interleukin-6 (IL-6), C-reactive protein (CRP), and interleukin 1-beta. Cognitive domains included memory, executive function, and attention/information processing. MRI measures included white matter hyperintensity volumes (WMH) and quantification of white matter integrity in areas outside WMHs via DTI-derived fractional anisotropy (FA) and mean diffusivity, as well as multi-component relaxometry derived myelin water fraction (MWF). RESULTS: Black and White participants did not differ on age, sex, or CVD risk. Separate linear regression models adjusting for relevant confounders revealed that higher IL-6 associated with lower executive function and higher CRP levels associated with lower FA and MWF. Stratified analyses revealed that these association were significant for Black participants only. DISCUSSION: These findings suggest that peripheral inflammation is inversely associated with select cognitive domains and white matter integrity (but not WMHs), particularly in older Black adults. It is important to consider race when investigating inflammatory associates of brain and behavior.
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(1) Background: There are currently very few interventions performed within a community setting that compare the effects of physical activity (PA) versus PA plus weight loss on cancer and chronic disease risk in older African Americans. Therefore, we investigated the impact of an 8 week (24 session) PA intervention compared to a PA plus weight loss intervention on fat mass, glucose metabolism, and markers of inflammation in older, overweight and obese African Americans. (2) Methods: Subjects were randomized to a PA (n = 83) or PA plus weight loss (n = 72) intervention that met three times weekly for 8 weeks. At baseline and post-intervention, anthropometrics, body composition, systemic inflammation (high-sensitivity C-reactive protein, tumor necrosis factor-α, and interleukin 6), fasting glucose, insulin and homeostasis model assessment-insulin resistance (HOMA-IR) were determined. (3) Results: Subjects had a mean age of 67 years (SD = 5.3) and were mostly women (88%). The PA plus weight loss group lost more total and visceral fat than the PA group (-4.0% vs. +0.6% and -4.1% vs. +3.7%, respectively, p < 0.01 for both). Changes in inflammation and glucose metabolism were similar between groups post-intervention. Within the PA plus weight loss group only, serum insulin and HOMA-IR decreased significantly. (4) Conclusions: PA combined with weight loss can decrease total and visceral fat mass and improve insulin sensitivity, confirming that these cancer- and chronic disease-related risk factors are influenced by relatively modest lifestyle changes in the short term.
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Exercício Físico/fisiologia , Osteoartrite/terapia , Sobrepeso/terapia , Redução de Peso/fisiologia , Programas de Redução de Peso/métodos , Negro ou Afro-Americano , Idoso , Biomarcadores/sangue , Glicemia/análise , Composição Corporal , Índice de Massa Corporal , Doença Crônica/prevenção & controle , Dieta Redutora/métodos , Terapia por Exercício/métodos , Jejum/sangue , Feminino , Humanos , Inflamação , Mediadores da Inflamação/sangue , Insulina/sangue , Resistência à Insulina , Gordura Intra-Abdominal/fisiopatologia , Estilo de Vida , Masculino , Neoplasias/etiologia , Neoplasias/prevenção & controle , Obesidade/sangue , Obesidade/complicações , Obesidade/terapia , Osteoartrite/sangue , Osteoartrite/complicações , Sobrepeso/sangue , Sobrepeso/complicações , Fatores de Risco , Resultado do TratamentoRESUMO
INTRODUCTION: Acute pancreatitis (AP) is a healthcare challenge with considerable mortality. Treatment is limited to supportive care, highlighting the need to investigate disease drivers and prognostic markers. Activin A is an established mediator of inflammatory responses, and its serum levels correlate with AP severity. We hypothesized that activin A is independent of body mass index (BMI) and is a targetable promoter of the AP inflammatory response. METHODS: We assessed whether BMI and serum activin A levels are independent markers to determine disease severity in a cohort of patients with AP. To evaluate activin A inhibition as a therapeutic, we used a cerulein-induced murine model of AP and treated mice with activin A-specific neutralizing antibody or immunoglobulin G control, both before and during the development of AP. We measured the production and release of activin A by pancreas and macrophage cell lines and observed the activation of macrophages after activin A treatment. RESULTS: BMI and activin A independently predicted severe AP in patients. Inhibiting activin A in AP mice reduced disease severity and local immune cell infiltration. Inflammatory stimulation led to activin A production and release by pancreas cells but not by macrophages. Macrophages were activated by activin A, suggesting activin A might promote inflammation in the pancreas in response to injury. DISCUSSION: Activin A provides a promising therapeutic target to interrupt the cycle of inflammation and tissue damage in AP progression. Moreover, assessing activin A and BMI in patients on hospital admission could provide important predictive measures for screening patients likely to develop severe disease.
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Ativinas/metabolismo , Anti-Inflamatórios/farmacologia , Pâncreas/patologia , Pancreatite/diagnóstico , Índice de Gravidade de Doença , Ativinas/antagonistas & inibidores , Ativinas/sangue , Ativinas/imunologia , Animais , Anti-Inflamatórios/uso terapêutico , Biomarcadores/sangue , Biomarcadores/metabolismo , Índice de Massa Corporal , Linhagem Celular , Ceruletídeo/administração & dosagem , Ceruletídeo/toxicidade , Estudos de Coortes , Modelos Animais de Doenças , Avaliação Pré-Clínica de Medicamentos , Feminino , Humanos , Ativação de Macrófagos/imunologia , Macrófagos , Camundongos , Pâncreas/efeitos dos fármacos , Pâncreas/imunologia , Pancreatite/sangue , Pancreatite/tratamento farmacológico , Pancreatite/imunologia , Admissão do Paciente , Valor Preditivo dos TestesRESUMO
Pancreatic ductal adenocarcinoma (PDAC) is notorious for its poor survival and resistance to conventional therapies. PI3K signaling is implicated in both disease initiation and progression, and specific inhibitors of selected PI3K p110 isoforms for managing solid tumors are emerging. We demonstrate that increased activation of PI3K signals cooperates with oncogenic Kras to promote aggressive PDAC in vivo. The p110γ isoform is overexpressed in tumor tissue and promotes carcinogenesis via canonical AKT signaling. Its selective blockade sensitizes tumor cells to gemcitabine in vitro, and genetic ablation of p110γ protects against Kras-induced tumorigenesis. Diet/obesity was identified as a crucial means of p110 subunit up-regulation, and in the setting of a high-fat diet, p110γ ablation failed to protect against tumor development, showing increased activation of pAKT and hepatic damage. These observations suggest that a careful and judicious approach should be considered when targeting p110γ for therapy, particularly in obese patients.
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Carcinogênese/genética , Carcinoma Ductal Pancreático/genética , Classe Ib de Fosfatidilinositol 3-Quinase/genética , Hepatopatia Gordurosa não Alcoólica/patologia , Obesidade/complicações , Neoplasias Pancreáticas/genética , Animais , Antimetabólitos Antineoplásicos/farmacologia , Antimetabólitos Antineoplásicos/uso terapêutico , Carcinogênese/efeitos dos fármacos , Carcinoma Ductal Pancreático/tratamento farmacológico , Carcinoma Ductal Pancreático/patologia , Classe Ia de Fosfatidilinositol 3-Quinase/genética , Classe Ib de Fosfatidilinositol 3-Quinase/metabolismo , Desoxicitidina/análogos & derivados , Desoxicitidina/farmacologia , Desoxicitidina/uso terapêutico , Dieta Hiperlipídica/efeitos adversos , Modelos Animais de Doenças , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Resistencia a Medicamentos Antineoplásicos/genética , Ácidos Graxos Ômega-6/efeitos adversos , Feminino , Glucose/metabolismo , Humanos , Metabolismo dos Lipídeos , Fígado/patologia , Masculino , Camundongos Knockout , Hepatopatia Gordurosa não Alcoólica/etiologia , Hepatopatia Gordurosa não Alcoólica/metabolismo , Obesidade/etiologia , Obesidade/metabolismo , Pâncreas/patologia , Neoplasias Pancreáticas/tratamento farmacológico , Neoplasias Pancreáticas/patologia , Inibidores de Fosfoinositídeo-3 Quinase/farmacologia , Inibidores de Fosfoinositídeo-3 Quinase/uso terapêutico , Proteínas Proto-Oncogênicas c-akt/metabolismo , Proteínas Proto-Oncogênicas p21(ras)/genética , Proteínas Proto-Oncogênicas p21(ras)/metabolismo , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética , Regulação para Cima , GencitabinaRESUMO
Although survival has improved in recent years, the prognosis of patients with advanced pancreatic ductal adenocarcinoma (PDAC) remains poor. Despite substantial differences in anatomy, physiology, genetics, and metabolism, the overwhelming majority of preclinical testing relies on transgenic mice. Hence, while mice have allowed for tremendous advances in cancer biology, they have been a poor predictor of drug performance/toxicity in the clinic. Given the greater similarity of sus scrofa pigs to humans, we engineered transgenic sus scrofa expressing a LSL-KRASG12D-TP53R167H cassette. By applying Adeno-Cre to pancreatic duct cells in vitro, cells self-immortalized and established tumors in immunocompromised mice. When Adeno-Cre was administered to the main pancreatic duct in vivo, pigs developed extensive PDAC at the injection site hallmarked by excessive proliferation and desmoplastic stroma. This serves as the first large animal model of pancreatic carcinogenesis, and may allow for insight into new avenues of translational research not before possible in rodents.
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Carcinoma Ductal Pancreático/genética , Neoplasias Pancreáticas/genética , Proteínas Proto-Oncogênicas p21(ras)/genética , Sus scrofa/genética , Proteína Supressora de Tumor p53/genética , Adenocarcinoma/genética , Adenocarcinoma/patologia , Animais , Animais Geneticamente Modificados , Carcinoma Ductal Pancreático/patologia , Feminino , Humanos , Integrases , Camundongos SCID , Mutação , Neoplasias Experimentais , Ductos Pancreáticos/patologia , Neoplasias Pancreáticas/patologia , Microambiente Tumoral , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
OBJECTIVE: In this study, the associations between vitamin D, insulin sensitivity, and inflammation and their relationships with adipose tissue expression of vitamin D receptor (VDR) and inflammatory markers in women with morbid obesity were determined. METHODS: An oral glucose tolerance test prior to surgery was completed by healthy premenopausal women (n = 76) seeking bariatric surgery. Abdominal subcutaneous adipose tissue (SAT) and visceral adipose tissue (VAT) were collected during surgery. RESULTS: Approximately, 70% of our subjects were vitamin D sufficient or optimal, and 80% had normal glucose tolerance. No significant association between serum 25-hydroxyvitamin D [25(OH)D] with circulating inflammatory markers or insulin sensitivity was identified. In subjects with waist circumference of <139 cm (n = 42), log25(OH)D positively predicted VAT logIL-6 mRNA expression (P = 0.003). LogVDR expression was positively correlated with the expression of inflammatory markers in both SAT (logIL-1ß mRNA: r = 0.95, P < 0.0001; logTNF mRNA: r = 0.82, P < 0.0001) and VAT (logIL-1ß mRNA: r = 0.89, P < 0.0001; logTNF mRNA: r = 0.75, P < 0.0001). VAT logVDR expression positively predicted logHOMA-IR in non-African American subjects (P = 0.05). CONCLUSIONS: The beneficial effects of vitamin D on inflammation and insulin sensitivity were not supported by our findings. VDR does not appear to possess a protective effect in adipose tissue.
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Resistência à Insulina , Obesidade Mórbida/metabolismo , Vitamina D/análogos & derivados , Adulto , Feminino , Teste de Tolerância a Glucose , Humanos , Inflamação/metabolismo , Gordura Intra-Abdominal/metabolismo , Pessoa de Meia-Idade , Pré-Menopausa , Vitamina D/metabolismoRESUMO
Acute pancreatitis (AP), although most often a mild and self-limiting inflammatory disease, worsens to a characteristically necrotic severe acute pancreatitis (SAP) in about 20% of cases. Obesity, affecting more than one-third of American adults, is a risk factor for the development of SAP, but the exact mechanism of this association has not been identified. Coincidental with chronic low-grade inflammation, activation of the nucleotide-binding domain, leucine-rich containing family, pyrin-domain containing 3 (NLRP3) inflammasome increases with obesity. Lean mice genetically deficient in specific components of the NLRP3 inflammasome are protected from experimentally induced AP, indicating a direct involvement of this pathway in AP pathophysiology. We hypothesized that inhibition of the NLRP3 inflammasome with the sulfonylurea drug glyburide would reduce disease severity in obese mice with cerulein-induced SAP. Treatment with glyburide led to significantly reduced relative pancreatic mass and water content and less pancreatic damage and cell death in genetically obese ob/ob mice with SAP compared with vehicle-treated obese SAP mice. Glyburide administration in ob/ob mice with cerulein-induced SAP also resulted in significantly reduced serum levels of interleukin 6, lipase, and amylase and led to lower production of lipopolysaccharide-stimulated interleukin 1ß release in cultured peritoneal cells, compared with vehicle-treated ob/ob mice with SAP. Together, these data indicate involvement of the NLRP3 inflammasome in obesity-associated SAP and expose the possible utility of its inhibition in prevention or treatment of SAP in obese individuals.
Assuntos
Proteínas de Transporte/metabolismo , Glibureto/farmacologia , Hipoglicemiantes/farmacologia , Pancreatite/induzido quimicamente , Amilases/sangue , Amilases/metabolismo , Animais , Proteínas de Transporte/antagonistas & inibidores , Proteínas de Transporte/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Interleucina-1beta/metabolismo , Interleucina-6/sangue , Interleucina-6/metabolismo , Lipase/sangue , Lipase/metabolismo , Macrófagos Peritoneais/efeitos dos fármacos , Macrófagos Peritoneais/metabolismo , Camundongos , Camundongos Obesos , Proteína 3 que Contém Domínio de Pirina da Família NLR , Pancreatite/metabolismo , Pancreatite/patologiaRESUMO
Serum samples from 74 obese women were assayed for 25-hydroxyvitamin D [25(OH)D] concentrations using an automated immunoassay [Architect (Abbott)] and ELISA (Alpco Diagnostics), and results were compared with the LC/MS/MS reference method (Quest Diagnostics). The Architect values were significantly lower (mean difference: -13 nmol/L; 95% limits: -54; 28 nmol/L) and the ELISA values were significantly higher (mean difference: 24 nmol/L; 95% limits: -36; 84 nmol/L) than the LC/MSIMS values. The slope of the Passing-Bablok regression line relative to LC/MSIMS was 0.5 [95% confidence interval (CI): 0.41; 0.60] and 1.17 (95% CI: 0.87; 1.56) for Architect and ELISA, respectively, with an intercept of approximately 16 for both assays. Using 50 nmol/L as the cut-point for deficiency, Architect and ELISA misclassified 20 and 27% of the subjects, respectively. In subjects with low circulating 25-hydroxylated ergocalciferol [25(OH)D2] (<10 nmol/L), a Bland-Altman plot and Kappa statistics (Kappa = 0.73; 95% CI: 0.49-0.97) showed good agreement between Architect and LC/MS/MS. However, in subjects with high circulating 25(OH)D2 (>or=10 nmol/L), Architect demonstrated poor agreement (Kappa = 0.40; 95% CI: 0.16-0.65). ELISA demonstrated a higher degree of overestimation in women with minimal 25(OH)D2 than those with high 25(OH)D2, suggesting that ELISA overestimates 25-hydroxylated cholecalciferol [25(OH)D3] but underestimates 25(OH)D2.
Assuntos
Vitamina D/sangue , Adulto , Cromatografia Líquida , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Espectrometria de Massas em Tandem , Vitamina D/análogos & derivadosRESUMO
OBJECTIVE: Currently, there is no therapy for severe acute pancreatitis (AP) except for supportive care. The lipase inhibitor orlistat, the peroxisome proliferator-activated receptor γ agonist rosiglitazone, and the chemokine receptor 2 antagonists attenuate the severity of AP in rodents if administered before or at the time of induction of pancreatitis. However, it is unknown whether these treatments are effective if administered therapeutically after induction of pancreatitis. METHODS: Male C57BL6 mice with diet-induced obesity received 2 injections of mrIL-12 (150 ng per mouse) and mrIL-18 (750 ng per mouse) intraperitoneally at 24-hour intervals. The mice were injected 2, 24, and 48 hours after the second injection of IL-12 + IL-18 with orlistat (2 mg per mouse), rosiglitazone (0.4 mg per mouse), RS102895 (0.3 mg per mouse), or vehicle (20 µL of DMSO and 80 µL of canola oil) and euthanized after 72 hours. RESULTS: Orlistat decreased intra-abdominal fat necrosis compared with vehicle (P < 0.05). However, none of the drug treatments produced significant decreases in pancreatic edema, acinar necrosis, or intrapancreatic fat necrosis. CONCLUSIONS: Drugs previously shown to improve the severity of AP when given before or at the time of induction of pancreatitis failed to do so when administered therapeutically in the IL-12 + IL-18 model.
Assuntos
Benzoxazinas/farmacologia , Lactonas/farmacologia , Obesidade/tratamento farmacológico , Pancreatite/tratamento farmacológico , Piperidinas/farmacologia , Tiazolidinedionas/farmacologia , Doença Aguda , Animais , Fármacos Antiobesidade/administração & dosagem , Fármacos Antiobesidade/farmacologia , Benzoxazinas/administração & dosagem , Interleucina-12 , Interleucina-18 , Lactonas/administração & dosagem , Masculino , Camundongos Endogâmicos C57BL , Obesidade/induzido quimicamente , Orlistate , Pancreatite/induzido quimicamente , Piperidinas/administração & dosagem , Receptores CCR2/antagonistas & inibidores , Rosiglitazona , Índice de Gravidade de Doença , Tiazolidinedionas/administração & dosagem , Falha de Tratamento , Vasodilatadores/administração & dosagem , Vasodilatadores/farmacologiaRESUMO
Obesity and type 2 diabetes are associated with increased production of Galectin-3 (Gal-3), a protein that modulates inflammation and clearance of glucose adducts. We used Lean and Diet-induced Obese (DIO) WT and Gal-3 KO mice to investigate the role of Gal-3 in modulation of adiposity, glucose metabolism and inflammation. Deficiency of Gal-3 lead to age-dependent development of excess adiposity and systemic inflammation, as indicated by elevated production of acute-phase proteins, number of circulating pro-inflammatory Ly6C(high) monocytes and development of neutrophilia, microcytic anemia and thrombocytosis in 20-week-old Lean and DIO male Gal-3 KO mice. This was associated with impaired fasting glucose, heightened response to a glucose tolerance test and reduced adipose tissue expression of adiponectin, Gal-12, ATGL and PPARγ, in the presence of maintained insulin sensitivity and hepatic expression of gluconeogenic enzymes in 20-week-old Gal-3 KO mice compared to their diet-matched WT controls. Expression of PGC-1α and FGF-21 in the liver of Lean Gal-3 KO mice was comparable to that observed in DIO animals. Impaired fasting glucose and altered responsiveness to a glucose load preceded development of excess adiposity and systemic inflammation, as demonstrated in 12-week-old Gal-3 KO mice. Finally, a role for the microflora in mediating the fasting hyperglycemia, but not the excessive response to a glucose load, of 12-week-old Gal-3 KO mice was demonstrated by administration of antibiotics. In conclusion, Gal-3 is an important modulator of glucose metabolism, adiposity and inflammation.
Assuntos
Adiposidade/fisiologia , Galectina 3/deficiência , Glucose/metabolismo , Inflamação/metabolismo , Adiponectina/genética , Adiponectina/metabolismo , Adiposidade/genética , Animais , Dieta , Fatores de Crescimento de Fibroblastos/genética , Fatores de Crescimento de Fibroblastos/metabolismo , Galectina 3/genética , Galectina 3/metabolismo , Glucose/genética , Teste de Tolerância a Glucose/métodos , Inflamação/genética , Insulina/genética , Insulina/metabolismo , Resistência à Insulina/genética , Lipase/genética , Lipase/metabolismo , Fígado/metabolismo , Camundongos , Camundongos Knockout , Camundongos Obesos , Monócitos/metabolismo , Obesidade/genética , Obesidade/metabolismo , PPAR gama/genética , PPAR gama/metabolismo , Coativador 1-alfa do Receptor gama Ativado por Proliferador de Peroxissomo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismoRESUMO
OBJECTIVE: Galectins (Gal) exert many activities, including regulation of inflammation and adipogenesis. We evaluated modulation of Gal-1, -3, -9 and -12 in visceral (VAT) and subcutaneous (SAT) adipose tissue in mice. DESIGN AND METHODS: We used two mouse models of obesity, high-fat diet induced obesity (DIO) and ob/ob mice. We also evaluated the response of Gal-1 KO mice to DIO. RESULTS: Both age and diet modulated expression of galectins, with DIO mice having higher serum Gal-1 and Gal-3 versus lean mice after 13-17 weeks of high-fat diet. In DIO mice there was a progressive increase in expression of Gal-1 and Gal-9 in SAT, whereas Gal-3 increased in both VAT and SAT. Expression of Gal-12 declined over time in VAT of DIO mice, similar to adiponectin. Obesity lead to increased production of Gal-1 in adipocytes, whereas the increased Gal-3 and Gal-9 of obesity mostly derived from the stromovascular fraction. Expression of Gal-12 was restricted to adipocytes. There was increased production of Gal-3 and Gal-9, but not Gal-1, in CD11c(-) and CD11c(+) macrophages from VAT of DIO versus lean mice. Expression of Gal-1, -3 and -12 in VAT and SAT of ob/ob mice followed a trend comparable to DIO mice. Rosiglitazone reduced serum Gal-1, but not Gal-3 and modulated expression of Gal-3 in VAT and Gal-9 and Gal-12 in SAT of DIO mice. High-fat feeding lead to increased adiposity in Gal-1 KO versus WT mice, with loss of correlation between leptin and adiposity and no alterations in glucose and insulin levels. CONCLUSIONS: Obesity leads to differential modulation of Gal-1, 3, 9 and 12 in VAT and SAT, with Gal-1 acting as a modulator of adiposity.