CRISPR-on system for the activation of the endogenous human INS gene.

Advances in the field of epigenetics have allowed the design of new therapeutic strategies to address complex diseases such as type 1 diabetes (T1D). Clustered regularly interspaced short palindromic repeats (CRISPR)-on is a novel and powerful RNA-guided transcriptional activator system that can turn on specific gene expression; however, it remains unclear whether this system can be widely used or whether its use will be restricted depending on cell types, methylation promoter statuses or the capacity to modulate chromatin state. Our results revealed that the CRISPR-on system fused with transcriptional activators (dCas9-VP160) activated endogenous human INS, which is a silenced gene with a fully methylated promoter. Similarly, we observed a synergistic effect on gene activation when multiple single guide RNAs were used, and the transcriptional activation was maintained until day 21. Regarding the epigenetic profile, the targeted promoter gene did not exhibit alteration in its methylation status but rather exhibited altered levels of H3K9ac following treatment. Importantly, we showed that dCas9-VP160 acts on patients' cells in vitro, particularly the fibroblasts of patients with T1D.

[Nutritional assessment of patients candidates for waiting list simultaneous kidney pancreas transplantation]. / Evaluación nutricional de pacientes candidatos a lista de espera de trasplante renopancreático.

BACKGROUND: Simultaneous Kidney Pancreas transplantation is currently the treatment of choice for patients with type 1 diabetes ERD advanced. Scientific studies on the therapeutic benefits of this guarantee acceptance of it as the most suitable for patients. The independence of dialysis and insulin therapy after the transplant represents a significant increase in the quality of life for these patients. In recent years, the importance of nutritional assessment in patients with chronic diseases has increased. There is increasing evidence that malnutrition is associated with an increased risk of morbidity and mortality after surgery. Given the therapeutic choice for these patients and the importance of nutritional status as an independent risk factor for post-transplant complications objective of our work is to describe the nutritional status of patients who are candidates for transplant through objective and subjective parameters of evaluation nutritional. MATERIALS AND METHODS: We included 45 patients with type 1 diabetes ERD interned in the Hospital Italiano of Buenos Aires for assessing pre Transplantation during the period June 2007-June 2008. They collected data through a sheet produced by themselves. As parameter anthropometric was calculated body mass index (BMI) by the formula Quetelet (post dialysis Weight (kg) / height 2 (m)) and was ranked as the ranges proposed by the committee of experts from WHO. As a subjective argument took place the Subjective Global Valuation (VGS), which ranked patients in well-nourished (A), mild malnourished (B), moderately malnourished (C) and severely malnourished (D). As biochemical parameters of the data was recorded serum albumin (g/dl) to hospitalizations and this data was obtained from the patient's medical history. RESULTS: The value of the average BMI of the total population was 21.83+/-2.65. According to the sex values were: 21.83+/-2.39 and 21.82+/-2.95 for male and female respectively. The nutritional status according to this indicator was anthropometric Normal in 89% of cases. The 6.66% of the cases submitted 13.33% pre malnutrition and obesity. We found no cases with obesity. Taking into account the albumin 47.45% of patients presented moderate risk of morbidity and mortality. According to GSV 62.22% were classified as well-nourished (Class A). It was the equitable proportion of women and men 33.33% were classified as mild malnourished (Class B). Only 2 cases (8.88%) had moderate malnutrition (Class C). None presented malnutrition Graves (Class C). CONCLUSION: In our study we can see that patients are candidates for waiting list while normal BMI have presented evidence of nutritional risk when one takes into account the VGS and the average value of albumin.

Evaluación nutricional de pacientes candidatos a lista de espera de trasplante renopancreático / Nutritional assessment of patients candidates for waiting list simultaneous kidney pancreas transplantation

Introducción: El trasplante renopancreático es en la actualidad el tratamiento elegido para los pacientes diabéticos tipo 1 con ERD avanzada. Estudios científicos sobre los beneficios de esta terapéutica avalan la aceptación de la misma como lo más adecuado para los pacientes. La independencia de diálisis y de tratamiento insulínico luego del trasplante representa un importante incremento en la calidad de vida de esos pacientes. En los últimos años, la importancia de la evaluación nutricional en pacientes con enfermedades crónicas ha aumentado. Existe cada vez más evidencia científica que la malnutrición esta asociada a un riesgo mayor de morbilidad y mortalidad post intervención quirúrgica. Teniendo en cuenta la terapéutica elegida para estos pacientes y la importancia del estado nutricional como factor predictor independiente de riesgo de complicaciones post trasplante es objetivo de nuestro trabajo describir el estado nutricional de los pacientes candidatos a trasplante renopancreatico a través de parámetros objetivos y subjetivos de valoración nutricional. Materiales y métodos: Se incluyeron 45 pacientes Diabéticos tipo 1 con ERD internados en el Hospital Italiano de Buenos Aires para evaluación pre trasplante renopancreático durante el período Junio 2007-Junio 2008. Se recolectaron los datos mediante una ficha de elaboración propia. Como parámetro antropométrico se calculó el Índice de Masa corporal (IMC) mediante la fórmula de Quetelet (Peso post diálisis (kg)/talla 2 (mts)) y se clasificó según los rangos propuestos por el comité de Expertos de la OMS. Como parámetro subjetivo se realizo la Valoración Global Subjetiva (VGS) que clasificó a los pacientes en Bien nutrido (A), desnutrido leve (B), desnutrido moderado (C) y desnutrido severo (D). Como parámetros bioquímicos se registró el dato de albúmina sérica (g/dl) al ingreso hospitalario y este dato se obtuvo de la historia clínica del paciente. Resultados: El valor de la media para IMC del total de la población fue de 21,83 ± 2,65. Según el sexo los valores fueron: 21,83 ± 2,39 y de 21,82 ± 2,95 para el sexo mascureprelino y femenino respectivamente. El estado nutricional según este indicador antropométrico fue Normal en el 89% de los casos. El 6,66% de los casos presentó desnutrición y 13,33% pre obesidad. No se encontraron casos con Obesidad. Teniendo en cuenta la albúmina el 47,45% de los pacientes presenta riesgo moderado de morbimortalidad. Según la VGS el 62,22% se clasificaron como bien nutridos (Clase A). Fue equitativa la proporción de mujeres y de hombres. 33,33% se clasificaron como desnutridos leve (Clase B). Sólo 2 casos (8,88%) presentaron desnutrición moderada (Clase C). Ninguno presentó desnutrición Graves (Clase C). Conclusión: En nuestro estudio se puede observar que los pacientes candidatos a lista de espera si bien presentan valores de IMC dentro de la clasificación normal presentan indicios de riesgo nutricional cuando se tienen en cuenta la VGS y el valor medio de albúmina (AU)

Background: Simultaneous Kidney Pancreas transplantation is currently the treatment of choice for patients with type 1 diabetes ERD advanced. Scientific studies on the therapeutic benefits of this guarantee acceptance of it as the most suitable for patients. The independence of dialysis and insulin therapy after the transplant represents a significant increase in the quality of life for these patients. In recent years, the importance of nutritional assessment in patients with chronic diseases has increased. There is increasing evidence that malnutrition is associated with an increased risk of morbidity and mortality after surgery. Given the therapeutic choice for these patients and the importance of nutritional status as an independent risk factor for post-transplant complications objective of our work is to describe the nutritional status of patients who are candidates for transplant through objective and subjective parameters of evaluation nutritional. Materials and methods: We included 45 patients with type 1 diabetes ERD interned in the Hospital Italiano of Buenos Aires for assessing pre Transplantation during the period June 2007-June 2008.Se collected data through a sheet produced by themselves. As parameter anthropometric was calculated body mass index (BMI) by the formula Quetelet (post dialysis Weight (kg) / height 2 (m)) and was ranked as the ranges proposed by the committee of experts from WHO. As a subjective argument took place the Subjective Global Valuation (VGS), which ranked patients in well-nourished (A), mild malnourished (B), moderately malnourished (C) and severely malnourished (D). As biochemical parameters of the data was recorded serum albumin (g / dl) to hospitalizations and this data was obtained from the patient's medical history. Results: The value of the average BMI of the total population was 21.83 ± 2.65. According to the sex values were: 21.83 ± 2.39 and 21.82 ± 2.95 for male and female respectively. The nutritional status according to this indicator was anthropometric Normal in 89% of cases. The 6.66% of the cases submitted 13.33% pre malnutrition and obesity. We found no cases with obesity. Taking into account the albumin 47.45% of patients presented moderate risk of morbidity and mortality. According to GSV 62.22% were classified as well-nourished (Class A). It was the equitable proportion of women and hombres.33, 33% were classified as mild malnourished (Class B). Only 2 cases (8.88%) had moderate malnutrition (Class C). None presented malnutrition Graves (Class C). Conclussion: In our study we can see that patients are candidates for waiting list while normal BMI have presented evidence of nutritional risk when one takes into account the VGS and the average value of albumin (AU)

Frequent loss of heterozygosity at chromosome 3p14.2-3p21 in human pancreatic islet cell tumours.

OBJECTIVE: Pancreatic islet betacell tumours occur either sporadically or as part of inherited neoplastic syndromes, most commonly multiple endocrine neoplasia (MEN) type 1. Recently, a transgenic mouse model has been established in which the expression of the SV40 large T antigen was targeted to betacells by the rat insulin promoter, leading to the development of multiple pancreatic betacell tumours. In the advanced stages of tumour evolution, these tumours exhibited a high prevalence of loss of heterozygosity (LOH) on mouse chromosomes 9 and 16, at regions syntenic with regions 3q, 3p21, 6q12, 15q24 and 22q of the human genome. DESIGN: Loss of heterozygosity in human islet cell tumours was analysed in a PCR based approach at regions of the human genome syntenic with the mouse loci linked to pancreatic betacell tumours as well as the MEN1 gene on chromosome 11q13. These included 35 microsatellite markers in the human chromosomal regions 3q, 3p21, 6q12, 11q13, 15q24 and 22q. PATIENTS: 21 patients diagnosed with insulinoma were analysed. Histologically, 16 tumours were benign, while 5 were malignant insulinomas. RESULTS: Thirteen of 21 (62%) tumours were found to have loss of genetic material on chromosome 3. The shortest region of overlap implicated a deletion at 3p14.2-3p21 region, corresponding to the marker D3S1295. We did not detect a substantial frequency of LOH in the other syntenic regions, except for the region of MEN 1 gene on 11q13 found to be deleted in 6 (29%) cases, including 3 of 4 tumours from MEN 1 families. Deletions of 3p14. 2-3p21 were observed in 8 of 15 (53%) benign tumours, and in 5 of 6 (83%) malignant neoplasms. CONCLUSIONS: These results indicate the high frequency of 3p14.2-3p21 deletions in human pancreatic betacell neoplasms. These finding suggest the presence of a tumour suppressor gene in this region, that may be important in the microevolution of these tumours towards malignancy.

[Kidney and pancreas transplantation: initial experience at a single transplant center in Argentina]. / Trasplante renopancrático. Experiencia inicial en un centro de trasplante en Argentina.

After more than 10,000 cases reported all over the world until 1998, simultaneous kidney and pancreas transplantation has become a safe clinical practice, and it may probably represent the best treatment available for diabetic patients in end-stage renal disease. Here we present our results after 12 cadaveric pancreas transplants (8 whole organ, and 4 islet transplants), performed on insulin-dependent diabetic patients. Eleven of these patients received a kidney simultaneously, and one of them required a kidney retransplantation. All vascularised pancreatic grafts were positioned intraperitoneally, anastomosed to the iliac vessels, and bladder drained. One year patient, whole pancreas, and kidney survival rates were 86%, 86% and 71%, respectively. All of these patients remain insulin and dialysis-free, the longest for 37 months. Islets for transplantation were obtained from single cadaveric donors. Fresh, unpurified cells were transplanted intraperitoneally by laparoscopy (equivalent islet yields: 3 x 10(5), 4 x 10(5), 1 x 10(6) and 5 x 10(5)). None of the islet recipients resulted insulin-independent but they all reduced daily requirements in about 40%, with better metabolic control (mean HbA1c pretransplant 9.4 +/- 1.8, vs 7.9 +/- 1.6 posttransplant). One kidney graft was lost due to venous thrombosis. Simultaneous kidney and pancreas transplantation offers the diabetic patient in end-stage renal disease a chance of independence both from dialysis and exogenous insulin. Whole pancreas transplantation has better functional outcome than islet transplantation. Nevertheless, for those diabetic patients who do not meet the criteria to receive a vascularised graft, pancreatic cells may still improve carbohydrate metabolism with minor surgical risk.

Trasplante renopancrático. Experiencia inicial en un centro de trasplante en Argentina. / [Kidney and pancreas transplantation: initial experience at a single transplant center in Argentina]

After more than 10,000 cases reported all over the world until 1998, simultaneous kidney and pancreas transplantation has become a safe clinical practice, and it may probably represent the best treatment available for diabetic patients in end-stage renal disease. Here we present our results after 12 cadaveric pancreas transplants (8 whole organ, and 4 islet transplants), performed on insulin-dependent diabetic patients. Eleven of these patients received a kidney simultaneously, and one of them required a kidney retransplantation. All vascularised pancreatic grafts were positioned intraperitoneally, anastomosed to the iliac vessels, and bladder drained. One year patient, whole pancreas, and kidney survival rates were 86

, 86

and 71

, respectively. All of these patients remain insulin and dialysis-free, the longest for 37 months. Islets for transplantation were obtained from single cadaveric donors. Fresh, unpurified cells were transplanted intraperitoneally by laparoscopy (equivalent islet yields: 3 x 10(5), 4 x 10(5), 1 x 10(6) and 5 x 10(5)). None of the islet recipients resulted insulin-independent but they all reduced daily requirements in about 40

, with better metabolic control (mean HbA1c pretransplant 9.4 +/- 1.8, vs 7.9 +/- 1.6 posttransplant). One kidney graft was lost due to venous thrombosis. Simultaneous kidney and pancreas transplantation offers the diabetic patient in end-stage renal disease a chance of independence both from dialysis and exogenous insulin. Whole pancreas transplantation has better functional outcome than islet transplantation. Nevertheless, for those diabetic patients who do not meet the criteria to receive a vascularised graft, pancreatic cells may still improve carbohydrate metabolism with minor surgical risk.

[A new methodology for the preoperative localization of occult insulinomas]. / Nueva metodología para la localizacion preoperatoria de insulinomas ocultos.

UNLABELLED: In patients with hypoglycemic syndrome, preoperative localization of the insulinoma highly contributes to surgical removal. When the ultrasonography, computed tomography, magnetic resonance and pancreatic angiography fail to visualize the tumor, they are called occult insulinomas (OI). In this paper we describe the results of a new diagnostic method to localize OI, performed in 5 patients with hypoglycemic syndrome secondary to endogenous hyperinsulinism. In four out of five patients, computed tomography, magnetic resonance and angiography failed to show any tumor. In just one single case, these imaging methods showed the pancreatic tumor. All patients were studied by selective intraarterial pancreatic stimulation (SIPS): a) infusion of calcium gluconate (0.025 mEq/kg) in each artery that supplies the pancreas: gastroduodenal, superior mesenteric and splenic arteries as well as the hepatic artery; b) insulin venous sampling in the right supra-hepatic vein at 30 and 60 seconds after arterial stimulation (in one patient an additional sample at 90 seconds was obtained). The study was considered pathologic when the gradient (basal vs post-stimulus) increased at least 100%. RESULTS: In all five patients a pathological gradient was found. The suspected preoperative localization of the tumor was confirmed at surgery in four cases. The anatomopathologic examination revealed insulinoma in four cases and malignant insulinoma in the remaining. It is concluded that the results of this preliminary experience show the usefulness of SIPS in the preoperative localization of occult insulinomas.