There is an urgency to find new treatments for the devastating epidemic of diabetes. Pancreatic β-cells viability and function are impaired in the two most common forms of diabetes, type 1 and type ...2. Regeneration of pancreatic β-cells has been proposed as a potential therapy for diabetes. In a preliminary study, we screened a collection of marine products for β-cell proliferation. One unique compound (epoxypukalide) showed capability to induce β-cell replication in the cell line INS1 832/13 and in primary rat cell cultures. Epoxypukalide was used to study β-cell proliferation by (3)Hthymidine incorporation and BrdU incorporation followed by BrdU/insulin staining in primary cultures of rat islets. AKT and ERK1/2 signalling pathways were analyzed. Cell cycle activators, cyclin D2 and cyclin E, were detected by western-blot. Apoptosis was studied by TUNEL and cleaved caspase 3. β-cell function was measured by glucose-stimulated insulin secretion. Epoxypukalide induced 2.5-fold increase in β-cell proliferation; this effect was mediated by activation of ERK1/2 signalling pathway and upregulation of the cell cycle activators, cyclin D2 and cyclin E. Interestingly, epoxypukalide showed protection from basal (40% lower versus control) and cytokine-induced apoptosis (80% lower versus control). Finally, epoxypukalide did not impair β-cell function when measured by glucose-stimulated insulin secretion. In conclusion, epoxypukalide induces β-cell proliferation and protects against basal and cytokine-mediated β-cell death in primary cultures of rat islets. These findings may be translated into new treatments for diabetes.
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Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
There is an urgency to find new treatments for the devastating epidemic of diabetes. Pancreatic beta-cells viability and function are impaired in the two most common forms of diabetes, type 1 and ...type 2. Regeneration of pancreatic beta-cells has been proposed as a potential therapy for diabetes. In a preliminary study, we screened a collection of marine products for beta-cell proliferation. One unique compound (epoxypukalide) showed capability to induce beta-cell replication in the cell line INS1 832/13 and in primary rat cell cultures. Epoxypukalide was used to study beta-cell proliferation by .sup.3 Hthymidine incorporation and BrdU incorporation followed by BrdU/insulin staining in primary cultures of rat islets. AKT and ERK1/2 signalling pathways were analyzed. Cell cycle activators, cyclin D2 and cyclin E, were detected by western-blot. Apoptosis was studied by TUNEL and cleaved caspase 3. beta-cell function was measured by glucose-stimulated insulin secretion. Epoxypukalide induced 2.5-fold increase in beta-cell proliferation; this effect was mediated by activation of ERK1/2 signalling pathway and upregulation of the cell cycle activators, cyclin D2 and cyclin E. Interestingly, epoxypukalide showed protection from basal (40% lower versus control) and cytokine-induced apoptosis (80% lower versus control). Finally, epoxypukalide did not impair beta-cell function when measured by glucose-stimulated insulin secretion. In conclusion, epoxypukalide induces beta-cell proliferation and protects against basal and cytokine-mediated beta-cell death in primary cultures of rat islets. These findings may be translated into new treatments for diabetes.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
AIM To analyze the diagnostic yield(DY), therapeutic impact(TI) and safety of capsule endoscopy(CE).METHODS This is a multi-centre, observational, analytical, retrospective study. A total of 163 ...patients with suspicion of celiac disease(CD)(mean age = 46.4 ± 17.3 years, 68.1% women) who underwent CE from 2003 to 2015 were included. Patients were divided into four groups: seronegative CD with atrophy(Group-Ⅰ, n = 19), seropositive CD without atrophy(Group-Ⅱ, n = 39), contraindication to gastroscopy(Group-Ⅲ, n = 6), seronegative CD without atrophy, but with a compatible context(Group-Ⅳ, n = 99). DY, TI and the safety of CE were analysed.RESULTS The overall DY was 54% and the final diagnosis was villous atrophy(n = 65, 39.9%), complicated CD(n = 12, 7.4%) and other enteropathies(n = 11, 6.8%; 8 Crohn’s). DY for groups Ⅰ to Ⅳ was 73.7%, 69.2%, 50% and 44.4%, respectively. Atrophy was located in duodenum in 24 cases(36.9%), diffuse in 19(29.2%), jejunal in 11(16.9%), and patchy in 10 cases(15.4%). Factors associated with a greater DY were positive serology(68.3% vs 49.2%, P = 0.034) and older age(P = 0.008). On the other hand, neither sex nor clinical presentation, family background, positive histology or HLA status were associated with DY. CE resultschanged the therapeutic approach in 71.8% of the cases. Atrophy was associated with a greater TI(92.3% vs 45.3%, P < 0.001) and 81.9% of the patients responded to diet. There was one case of capsule retention(0.6%). Agreement between CE findings and subsequent histology was 100% for diagnosing normal/other conditions, 70% for suspected CD and 50% for complicated CD.CONCLUSION CE has a high DY in cases of suspicion of CD and it leads to changes in the clinical course of the disease. CE is safe procedure with a high degree of concordance with histology and it helps in the differential diagnosis of CD.
El dengue es una de las enfermedades virales más frecuente en el mundo, que causa cifras importantes de morbilidad y mortalidad en la región de América, afectando a varios países entre ellos ...Venezuela. Por tal motivo nos planteamos diagnosticar los conocimientos, actitudes y prácticas sobre dengue, aplicando una encuesta, en 4 comunidades, de 4 municipios del estado Nueva Esparta: Achípano (Mariño); Guacuco (Arismendi); El Valle (García) y Pampatar (Maneiro). Los resultados destacan la presencia de menores de 20 años y de sexo femenino; la mayoría de los jefes de familia tiene educación básica completa; un alto porcentaje ha escuchado sobre dengue. En lo que respecta al nivel de conocimiento, se encontró bajo en Guacuco, medio en Achípano y Pampatar y alto en El valle. Los síntomas más relacionados fueron: dolor de cabeza, muscular abdominal y vómitos; la mayoría reconoce que es una enfermedad producida por un virus y transmitida por mosquitos; existe desconocimiento sobre la intervención del consumo de agua y alimentos, así como la creencia de que todos los mosquitos son vectores; la mayoría de los encuestados reconoce que el vector es el mosquito patas blancas y que pica tanto de día como de noche; la mayoría reconoce que los mosquitos se reproducen en agua limpia. Con respecto a las actitudes, un alto porcentaje estuvo muy de acuerdo y de acuerdo con que el dengue es una enfermedad grave y corre el riesgo de enfermarse; consideran que eliminar los criaderos ayudará a prevenir la enfermedad y la comunidad puede participar. En lo que se refiere a las prácticas la mayoría almacena agua y la tapa adecuada mente; limpian semanalmente los recipientes y retiran la basura; muy bajo porcentaje se cuida personalmente, como usar mosquitero, repelente y ropa larga; la mayoría acudiría a un centro de salud para su tratamiento.
Abstract
Background: We hypothesized that the biphasic relapse pattern of TNBC could be explained by a limited number of activation patterns of signaling nodes. In addition, we sought to determine ...whether the hyperactive signaling nodes, distinguishing the cases with favorable vs adverse outcome, could be potential targets.
Methods: Training set of 34 frozen tumor samples divided in two sets, (A) 13 patients, relapsed in <4 years; (B) 21 patients relapse-free >12 years, (mimicking the percentage and relapse patterns of unselected TNBC, but paired for T, N, G and Ki67). TNBC cell lines: 7 indolent (no metastases in 60 weeks) and 3 aggressive, develop metastases and kill recipient mice in <4 months. Shotgun phosphoproteomics and TiO2-IMAC phosphopeptide enrichment coupled with mass spectrometry runs in a Orbitrap Elite Mass Spectrometer was performed. Spectra were processed with MaxQuant software. Differentially expressed phosphopeptides were obtained by applying linear models R limma package. Differential kinase activation driving the profiles segregating cured vs. relapsing cases was done using linear sequence motif analysis. The hyperactivated kinases were validated in an independent set of 113 consecutive TNBC cases with 12+ years of follow-up spotted in TMAs by using an in-house algorithm for immunohistochemistry coupled with computer-aided quantitation using an Ariol scanning, we took the kinases in the upper quartile (high activity). Survival analysis was performed with KM curves and log rank test; and Cox proportionate hazards model was used for multivariate models.
Results: 11405 phosphopeptides were identified and quantified in the training set. Supervised clustering of relapsed vs. cured cases showed that 161 and 541 peptides were significantly up-regulated in the A and B groups, respectively (FDR<0.15). After kinase-to-kinase co-linearity was ruled-out , gathering the high activity (upper quartile) of six kinases (a combined variable, herein Var1) showed statistically significant association with relapse, being these: PRKCE, pERK, c-KIT, CDK6, pP70S6K and pPNKP. Cox proportional hazards model of any of the six probes high (var1) vs rest: 9.9 vs. not reached years (P<0.001). Patients that had any of the 6 kinases high have 47% of chance to relapse (only 2 out of 42 relapsed patients have 0/6 active kinases) vs patients with Var1 negative, 7% of chance (29 patients out of 72 have 0/6 active kinases) we also observed constants patterns of activations in the different sets expressions of kinases. We considered the kinases at Var1 as a potentially targets and we developed a pharmacological in vitro assay, testing pairs inhibitors on 10 TNBC cell lines; 99.3% of the combinations were supra-additive.
Conclusion: High throughput p-proteomics allows a parsimonious segregation of early TNBC cases, easily detecting the cases with long-term cure vs the remaining while identifying potential therapeutic solutions for the patients falling in the adverse prognostic subgroups.
Citation Format: Sara Fernandez Gaitero, Ivana Zagorac, Jose Francisco Lopez-Acosta, Gonzalo Gomez-Lopez, David González Pisano, Javier Muñoz Peralta, Luis Manso, Soledad Alonso, Renske Penning, Maarten Altelaar, Albert JR Heck, Miguel Quintela-Fandino. Triple-negative breast cancer (TNBC) phosphoproteomics abstract. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1221. doi:10.1158/1538-7445.AM2017-1221
The correct image displays the red and green insulin staining, which is viewable at the following link: thumbnail Download: * PPT PowerPoint slide * PNG larger image * TIFF original image . (2013) ...Correction: Epoxypukalide Induces Proliferation and Protects against Cytokine-Mediated Apoptosis in Primary Cultures of Pancreatic β-Cells.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Activation of pancreatic β-cell proliferation has been proposed as an approach to replace reduced functional β-cell mass in diabetes. Quiescent fibroblasts exit from G0 (quiescence) to G1 through pRb ...phosphorylation mediated by cyclin C/cdk3 complexes. Overexpression of cyclin D1, D2, D3, or cyclin E induces pancreatic β-cell proliferation. We hypothesized that cyclin C overexpression would induce β-cell proliferation through G0 exit, thus being a potential therapeutic target to recover functional β-cell mass. We used isolated rat and human islets transduced with adenovirus expressing cyclin C. We measured multiple markers of proliferation: (3)Hthymidine incorporation, BrdU incorporation and staining, and Ki67 staining. Furthermore, we detected β-cell death by TUNEL, β-cell differentiation by RT-PCR, and β-cell function by glucose-stimulated insulin secretion. Interestingly, we have found that cyclin C increases rat and human β-cell proliferation. This augmented proliferation did not induce β-cell death, dedifferentiation, or dysfunction in rat or human islets. Our results indicate that cyclin C is a potential target for inducing β-cell regeneration.
Inflammation is a key event that is closely associated with the pathophysiology of frailty. The relationship of genetic polymorphisms into inflammatory cytokines with frailty remains poorly ...understood. The aim of this study was to investigate the association between VNTR polymorphisms of the
IL
-
4
and
IL
-
1RN
genes with the risk of frailty. We included a sample of 630 community-dwelling elderly aged 70 and older. Both
IL
-
4
and
IL
-
1RN
VNTR polymorphisms were genotyped by the polymerase chain reaction (PCR) method. Mean age was 77.7 years (SD = 6.0) and 52.5 % were women. The participants classified as frail were more likely to be older, had lower MMSE score (
p
< 0.001), and had more disability for IADL (
p
< 0.001) and ADL (
p
< 0.001). Genotypic and allelic frequencies for the
IL
-
4
VNTR polymorphism did not show significant differences between study groups (
p
> 0.05). However, we just observed a significant difference in the allelic frequencies for the A2 allele of the
IL
-
1RN
VNTR polymorphism between frail and nonfrail groups (OR 1.84, 95 % CI 1.08–3.12,
p
= 0.02). In addition, we analyzed the combined effect of the
IL
-
4
and
IL
-
1RN
VNTR polymorphisms and their possible association with frailty, where the combined
IL
-
4
low
–
IL
-
1Ra
high
genotype was identified as a marker of risk to frailty syndrome (OR 7.86, 95 % CI 1.83–33.69,
p
= 0.006). Our results suggest that both A2 allele and the combined
IL
-
4
low
–
IL
-
1Ra
high
genotype might be genetic markers of susceptibility to frailty in Mexican elderly.
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