Carolina Uribe Cruz

Platelet increases survival in a model of 90% hepatectomy in rats.

Ninety per cent hepatectomy in rodents is a model for acute liver failure. It has been reported that platelets have a strong effect enhancing liver regeneration, because of the production of several growth factors such as serotonin. The aim of this study was to investigate the role of microencapsulated platelets on 90% hepatectomy in rats.

Bone Marrow-Derived Mononuclear Cells Differentiate into Hepatocyte-Like Cells within Few Hrs without Fusion

Background: Cell therapy using Bone Marrow Mononuclear Cells (BMMC) has been shown as a potential treatment for liver diseases. BMMC can act by fusion, differentiation into hepatocyte-like cells and/or secretion of paracrine factors. Here, we used encapsulated BMMC in a model of Carbon Tetrachloride (CCl4)-induced acute liver injury to study in vivo and in vitro differentiation of BMMC. Methods: Both in vitro and in vivo studies were conducted in Wistar rats submitted to CCl4-induced acute liver injury. BMMC were isolated from Wistar rats and encapsulated in sodium alginate microcapsules. For in vivo experiments, animals received encapsulated BMMC 24 hrs after CCl4 administration and capsules were collected within 6, 24 and 48 hrs (tCCl4 group). For in vitro experiments, isolated hepatocytes from animals with CCl4-induced liver injury were co-cultured with encapsulated BMMC for 6 h (cCCl4 group). Control groups were not submitted to CCl4 administration. The content of intracellular lipid droplets in hepatocytes was used to evaluate liver injury. BMMC differentiation was assessed by RT-PCR for hepatic genes and ability to produce and secrete urea. Results: Liver damage was confirmed in CCl4 treated animals by the presence of intracellular lipid droplets in hepatocytes and the characteristic nutmeg aspect of the liver. Retrieved encapsulated BMMC from tCCl4 group expressed hepatocyte markers, such as Cytokeratin 18 and Albumin 48 hrs after treatment. On the other hand, BMMC from cCCl4 group showed Albumin expression 6 hrs after co-culture. Urea production was increased in BMMC from cCCl4 group but not in cControl. BMMC from tControl or cControl groups did not express hepatocyte markers at any time point. Conclusions: In this study we show that BMMC differentiate into hepatocyte-like cells in a short period of time both in vivo and in vitro. This differentiation is triggered by paracrine factors present only in injured liver.

The effects of anesthetic regimen in 90% hepatectomy in rats

PURPOSE To evaluate the influence of the anesthetic regimen on anesthetic recovery, survival, and blood glucose levels following a 90% partial hepatectomy in rats. METHODS Thirty adult male Wistar rats were divided into two groups according to their anesthetic regimens: intraperitoneal ketamine and xylazine or inhaled isoflurane. In order to prevent hypoglycemia, glucose was administered intraperitoneally and glucose (20%) was added to the drinking water. RESULTS Anesthetic recovery time was longer in the ketamine and xylazine group. The survival rate after 72 hours was lower (log rank=0.0001) in the ketamine and xylazine group (0.0%) than in the isoflurane group (26.7%). The blood glucose after six hours was lower (p=0.017) in the ketamine and xylazine group (63 ± 31.7 mg/dL) than in the isoflurane group (98 ± 21.2 mg/dL). The prolonged anesthesia recovery time associated with ketamine and xylazine decreased the survival rate and blood glucose levels after 90% hepatectomy. CONCLUSION Isoflurane anesthesia reduced the recovery time and incidence of hypoglycemia and increased the survival rate in the early hours, providing a therapeutic window that is suitable for experimental studies.

Salivary 17-α-Hydroxyprogesterone in Management of Congenital Adrenal Hyperplasia

The objective of this study was to standardize a radioimmunoassay for salivary 17-hydroxyprogesterone (17-OHP) and to determine its usefulness for monitoring therapy in 23 patients with congenital adrenal hyperplasia (CAH) due to 21-hydroxylase deficiency. The assay uses 0.5 ml of saliva incubated overnight with tritiated 17-OHP, at 4°C. The solvent utilised was ethyl ether. The assay is specific, precise and has a lower limit of sensitivity, 86 pmol/1. We found a positive correlation (r=0.77, pcO.OOOl) between 17-OHP concentrations in paired saliva and plasma samples from patients with CAH. The adequacy of steroid therapy was assessed on the basis of clinical findings, bone age, plasma testosterone and dehydroepiandrosterone sulfate over a 4-year period, when salivary and plasma levels were measured at 9.00 a.m. and 4.00 p.m. The 145 patient-periods were divided into 3 groups: adequately controlled (61.4%), overtreated (31%) and poorly controlled (7.6%). Individual salivary and plasma 17-OHP values were also matched to normal values for age and sex. When salivary 17-OHP values obtained from CAH groups at 9.00 a.m. were analysed an upper limit of 750 pmol/1 was established. Values above this limit were associated with

Gastrointestinal disorders and miglustat therapy: A case report

CO RR EC TE D P RO OF the juvenile forms by age of onset, rate of disease progression and age of death. There are no approved treatments for gangliosidoses. Biomarkers for evaluating disease phenotype, disease progression, and response to therapies would serve to facilitate the design and development of potential treatment of these diseases, but such biomarkers have not been identified. Increasing evidence from animal models and human cadavers suggests inflammatory mediators in the CNS play a role in disease pathology and progression. Hypothesis: The more rapid disease progression and more severe clinical phenotype of the infantile GM1and GM2-gangliosidosis relative to juvenile forms will coincide with levels of disease specific inflammatory mediators in the CNS. Methods: CSF and serum inflammatory markers were quantified by immunoassay in 8 children with infantile forms of gangliosidoses (including patients with Tay–Sachs disease, Sandhoff disease and GM1 gangliosidosis). Findings were compared to values from 4 patients with more slowly progressing forms of gangliosidoses (i.e., late-infantile, juvenile forms) and to values of 9 children with mucopolysaccharidosis (MPS) diseases. Results: Of 188 analytes assayed, elevated inflammatory markers occurredmore often in the CSF of infantile gangliosidosis patients when compared to the more slowly progressing forms of juvenile gangliosidosis and MPS disease. Five candidate biomarkers were distinguished by persistent elevation in the CSF of patients with the severe infantile phenotype: ENA-78, MCP-1, MIP-1α, MIP-1β, TNFR2. Additional candidates for CNS and serum inflammatory markers for infantile and juvenile phenotypes were found. Conclusion: This study identified candidate biomarkers of CNS inflammatory disease in infantile and juvenile gangliosidoses that are promising as markers for evaluating CNS disease progression, distinguishing infantile and juvenile phenotypes and monitoring response to future therapeutic interventions in the gangliosidoses. (Supported by Lysosomal Disease Network, NIH U54NS065768 and Pharmacotherapy for InheritedMetabolic Diseases Fellowship training program, GenzymeSanofi.)

The challenges of organizing an international course in Latin America

The Latin American School of Human and Medical Genetics (ELAG) is the main course of its kind in the genetics field in Latin America. Here we describe the main challenges regarding the organization of such event, including how we obtain funding and how we proceed with student selection. Thus, we aim to share our experience with other groups that intend to follow this format to create similar events in other areas in this region of the world.