Life after death? Exploring biochemical and molecular changes following organismal death in green turtles, Chelonia mydas (Linnaeus, 1758).

Green turtles, Chelonia mydas, have been included in biomonitoring efforts given its status as an endangered species. Many studies, however, rely on samples from stranded animals, raising the question of how death affects important biochemical and molecular biomarkers. The goal of this study was to investigate post mortem fluctuations in the antioxidant response and metabolism of carbohydrates in the liver of C. mydas. Liver samples were obtained from six green turtles which were submitted to rehabilitation and euthanized due to the impossibility of recovery. Samples were collected immediately after death (t = 0) and at various time intervals (1, 2, 3, 4, 5, 6, 12, 18 and 24 h post mortem), frozen in liquid nitrogen and stored at -80 °C. The activities of catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR) and glucose-6-phosphate dehydrogenase (G6PDH) were analyzed, as were the levels of lipid peroxidation, glycogen concentration, RNA integrity (RNA IQ) and transcript levels of carbonic anhydrase and pyruvate carboxylase genes. Comparison between post mortem intervals showed a temporal stability for all the biomarkers evaluated, suggesting that changes in biochemical and molecular parameters following green turtle death are not immediate, and metabolism may remain somewhat unaltered up to 24 h after death. Such stability may be associated with the overall lower metabolism of turtles, especially under an oxygen deprivation scenario such as organismal death. Overall, this study supports the use of biomarkers in sea turtles sampled within a period of 24 h post mortem for biomonitoring purposes, though it is recommended that post mortem fluctuations of particular biomarkers be evaluated prior to their application, given that proteins may show varying degrees of susceptibility to proteolysis.

Cyclosporine Versus Tacrolimus: Which Calcineurin Inhibitor Has Influence on Cytomegalovirus Infection in Cardiac Transplantation?

INTRODUCTION: Cytomegalovirus (CMV) infection is a relevant cause of morbidity and mortality in transplantation patients. Its major incidence is in the first year and viral replication is related to acute rejection, survival reduction, and graft vascular disease. OBJECTIVE: This study aims to evaluate retrospectively whether a high dose of calcineurin inhibitors correlates with CMV-positive polymerase chain reaction (PCR), need for treatment, and death in cardiac transplantation patients. METHODS: This is a case-control study including patients who underwent transplantation between 2014 and 2016. They were separated into two groups (positive or negative PCR) and evaluated for dosage serum levels of cyclosporine and tacrolimus. Patients were classified with adequate dose of immunosuppressant or high dose, and was analyzed that there was any association with those and positive CMV-PCR, need for treatment for CMV, and deaths. For statistical analysis, the Student t test was used for the quantitative variables and the Fisher's Exact Test for qualitative variables. To show CMV-free survival, the Kaplan-Meier curve was used. The level of significance was set at 5%. RESULTS: CMV-positive PCR in the sample was 72% for a total of 50 individuals. Positive PCR correlated with a high dose of calcineurin inhibitors in a statistically significant way (P = .002), as did a high dose of cyclosporine (P = .004); however, a high dose of tacrolimus had no such association (P = .17). When a high dose was assessed with a need for treatment, the chance of needing treatment increased more than eight times (P = .024; odds ratio = 8.25; 95% CI = 1.33 to 51.26), which was different from results found with high-dose tacrolimus (P = 1.0). However, no significant association was found in relation to deaths. CONCLUSIONS: Tacrolimus serum levels showed no association with CMV-PCR, which was different from serum cyclosporine, which showed association with CMV-PCR positivity, increasing the need for treatment approximately 8-fold, without association with death.

Cyclosporine Versus Tacrolimus: Which Calcineurin Inhibitor Has Influence on Cytomegalovirus Infection in Cardiac Transplantation?

INTRODUCTION: Cytomegalovirus (CMV) infection is a relevant cause of morbidity and mortality in transplantation patients. Its major incidence is in the first year and viral replication is related to acute rejection, survival reduction, and graft vascular disease. Objective. This study aims to evaluate retrospectively whether a high dose of calcineurin inhibitors correlates with CMV-positive polymerase chain reaction (PCR), need for treatment, and death in cardiac transplantation patients. Methods. This is a case-control study including patients who underwent transplantation between 2014 and 2016. They were separated into two groups (positive or negative PCR) and evaluated for dosage serum levels of cyclosporine and tacrolimus. Patients were classified with adequate dose of immunosuppressant or high dose, and was analyzed that there was any association with those and positive CMV-PCR, need for treatment for CMV, and deaths. For statistical analysis, the Student t test was used for the quantitative variables and the Fisher's Exact Test for qualitative variables. To show CMV-free survival, the Kaplan-Meier curve was used. The level of significance was set at 5%. Results. CMV-positive PCR in the sample was 72% for a total of 50 individuals. Positive PCR correlated with a high dose of calcineurin inhibitors in a statistically significant way (P » .002), as did a high dose of cyclosporine (P » .004); however, a high dose of tacrolimus had no such association (P » .17). When a high dose was assessed with a need for treatment, the chance of needing treatment increased more than eight times (P » .024; odds ratio » 8.25; 95% CI » 1.33 to 51.26), which was different from results found with high-dose tacrolimus (P » 1.0). However, no significant association was found in relation to deaths. Conclusions. Tacrolimus serum levels showed no association with CMV-PCR, which was different from serum cyclosporine, which showed association with CMV-PCR positivity, increasing the need for treatment approximately 8-fold, without association with death.

A segment of rbcL gene as a potential tool for forensic discrimination of Cannabis sativa seized at Rio de Janeiro, Brazil.

Cannabis sativa, known by the common name marijuana, is the psychoactive drug most widely distributed in the world. Identification of Cannabis cultivars may be useful for association to illegal crops, which may reveal trafficking routes and related criminal groups. This study provides evidence for the performance of a segment of the rbcL gene, through genetic signature, as a tool for identification for C. sativa samples apprehended by the Rio de Janeiro Police, Brazil. The PCR amplified and further sequenced the fragment of approximately 561 bp of 24 samples of C. sativa rbcL gene and showed the same nucleotide sequences, suggesting a possible genetic similarity or identical varieties. Comparing with other Cannabaceae family sequences, we have found 99% of similarity between the Rio de Janeiro sequence and three other C. sativa rbcL genes. These findings suggest that the fragment utilized at this study is efficient in identifying C. sativa samples, therefore, useful in genetic discrimination of samples seized in forensic cases.