Establishment of killer whale (Orcinus orca) primary fibroblast cell cultures and their transcriptomic responses to pollutant exposure.

Populations of killer whale (Orcinus orca) contain some of the most polluted animals on Earth. Yet, the knowledge on effects of chemical pollutants is limited in this species. Cell cultures and in vitro exposure experiments are pertinent tools to study effects of pollutants in free-ranging marine mammals. To investigate transcriptional responses to pollutants in killer whale cells, we collected skin biopsies of killer whales from the Northern Norwegian fjords and successfully established primary fibroblast cell cultures from the dermis of 4 out of 5 of them. Cells from the individual with the highest cell yield were exposed to three different concentrations of a mixture of persistent organic pollutants (POPs) that reflects the composition of the 10 most abundant POPs found in Norwegian killer whales (p,p'-DDE, trans-nonachlor, PCB52, 99, 101, 118, 138, 153, 180, 187). Transcriptional responses of 13 selected target genes were studied using digital droplet PCR, and whole transcriptome responses were investigated utilizing RNA sequencing. Among the target genes analysed, CYP1A1 was significantly downregulated in the cells exposed to medium (11.6 µM) and high (116 µM) concentrations of the pollutant mixture, while seven genes involved in endocrine functions showed a non-significant tendency to be upregulated at the highest exposure concentration. Bioinformatic analyses of RNA-seq data indicated that 13 and 43 genes were differentially expressed in the cells exposed to low and high concentrations of the mixture, respectively, in comparison to solvent control. Subsequent pathway and functional analyses of the differentially expressed genes indicated that the enriched pathways were mainly related to lipid metabolism, myogenesis and glucocorticoid receptor regulation. The current study results support previous correlative studies and provide cause-effect relationships, which is highly relevant for chemical and environmental management.

The clinical utility of D-dimer/platelet count ratio in pregnant women.

OBJECTIVE: We performed a retrospective study to assess the clinical utility of a new index, D-dimer/platelet count (DD/PLT) ratio, in discriminating preeclampsia from normal pregnancy and gestational hypertension during third trimester, compared to the biomarkers currently used, such as D-dimer (DD), platelet (PLT), lymphocyte (LIN) and neutrophil (NEU) counts, fibrinogen (FIB), PLT/NEU, NEU/LIN and PLT/LIN ratios. STUDY DESIGN: We retrospectively included 213 subjects. Of them, 163 and 50 were singleton pregnant and healthy non-pregnant women, respectively. Among pregnant women, 105 had normal pregnancy, 33 had gestational hypertension, and 25 had preeclampsia. RESULTS: Using Receiver Operating Curve (ROC) analysis, DD/PLT ratio showed significant higher area under the curve (AUC) (0.90; 95% confidence interval (CI) 0.84-0.95) in discriminating preeclampsia from normal pregnancy compared to those of DD, NEU, FIB, LIN, PLT/NEU, NEU/LIN and PLT/LIN ratios (p < .03). In discriminating preeclampsia from gestational hypertension, the DD/PLT AUC (0.90; 95% CI 0.79-0.96) was significantly higher than those of DD, NEU, FIB, LIN, NEU/LIN and PLT/LIN ratios (p < .03), and not statistically different from those of PLT (p = .22) and PLT/NEU ratio (p = .46). CONCLUSIONS: This study shows that DD/PLT ratio helps to discriminate preeclampsia from normal pregnancy and gestational hypertension. Large-scale studies are needed to verify its clinical usefulness, and to suggest more appropriate cutoff values for a widespread use.

The natural history of Peyronie's disease.

OBJECTIVE: Many physical or medical therapeutic approaches, systemic or local, have been suggested for treatment of Peyronie's disease. These approaches claim a discrete percentage of success in terms of clinical stabilization or improvement. The aim of our work was to evaluate the "natural history" of this disease. METHOD: 110 patients affected by Peyronie's disease have been observed for at least five years. At the first visit all patients maintained sexual activity not requiring surgical approach. No medical or physical treatments have been performed on these patients. All patients underwent twelve months follow up for at least 5 years evaluating the natural progression of the disease by means of ultrasound and clinical examination. We made statistical analysis (Odds ratio, P for trend) to check if there is association between clinical worsening requiring surgical treatment and: 1- age of patients, 2- diabetes, 3- presence of close relative pattern for diabetes and 4-Dupuytren contracture. RESULTS: Regarding curvature, number and size of fibrous plaques, a consistent tendency to stabilization has been observed in the group of patients above 50 years of age. 68% of the patients belonging to the group under 50 years showed a progressive worsening of the disease requiring surgical therapy, while in the other group only 31,5% of the patients required surgery. The statistical analysis confirmed that PD worsening is more probable in the group under 50 years of age (OR=3.5, CI:2-8) and in the patients affected by diabetes (age adjusted OR=6, CI:2-19). Statistical analysis has not shown significant differences in the evolution of Peyronie's disease regarding the presence of close relative pattern for diabetes and Dupuytren contracture. CONCLUSION: The patients presented a discrete tendency to spontaneous stabilization. The patients who are diagnosed under the fifties have a greater probability that the disease will worsen, requiring a surgical approach. The data regarding the natural history of Peyronie's disease in not treated patients should induce caution in awarding efficacy to new pharmacological and physical treatments.

The natural history of peyronie’s disease / Historia natural de la enfermedad de La Peyronie

Objetive: Many physical or medical therapeutic approaches, systemic or local, have been suggested for treatment of Peyronie's disease. These approaches claim a discrete percentage of success in terms of clinical stabilization or improvement. The aim of our work was to evaluate the «natural history» of this disease. Method: 110 patients affected by Peyronie's disease have been observed for at least five years. At the first visit all patients maintained sexual activity not requiring surgical approach. No medical or physical treatments have been performed on these patients. All patients underwent twelve months follow up for at least 5 years evaluating the natural progression of the disease by means of ultrasound and clinical examination. We made statistical analysis (Odds ratio, P for trend) to check if there is association between clinical worsening requiring surgical treatment and: 1- age of patients, 2- diabetes, 3- presence of close relative pattern for diabetes and 4-Dupuytren contracture. Results: Regarding curvature, number and size of fibrous plaques, a consistent tendency to stabilization has been observed in the group of patients above 50 years of age. 68% of the patients belonging to the group under 50 years showed a progressive worsening of the disease requiring surgical therapy, while in the other group only 31,5% of the patients required surgery. The statistical analysis confirmed that PD worsening is more probable in the group under 50 years of age (OR=3.5, CI:2-8) and in the patients affected by diabetes (age adjusted OR=6, CI:2-19). Statistical analysis has not shown significant differences in the evolution of Peyronie's disease regarding the presence of close relative pattern for diabetes and Dupuytren contracture. Conclusion: The patients presented a discrete tendency to spontaneous stabilization. The patients who are diagnosed under the fifties have a greater probability that the disease will worsen, requiring a surgical approach. The data regarding the natural history of Peyronie's disease in not treated patients should induce caution in awarding efficacy to new pharmacological and physical treatments (AU)

Objetivo: Para el tratamiento de la enfermedad de La Peyronie se han sugerido muchos abordajes terapéuticos físicos y médicos, sistémicos y locales. Estos tratamientos suponen un discreto porcentaje de éxitos en términos de estabilización clínica o mejoría. El objetivo de nuestro trabajo es evaluar la historia natural de esta enfermedad. Métodos: 110 pacientes con enfermedad de La Peyronie fueron observados durante al menos cinco años. En la primera visita todos los pacientes mantenían actividad sexual y no tenían indicación para un abordaje quirúrgico. No se hizo tratamiento médico físico en estos pacientes. Todos los pacientes fueron seguidos de anualmente durante al menos cinco años, evaluando la progresión natural de la enfermedad por medio de ultrasonidos y exploración física. Realizamos un estudio estadístico (Odds ratio, P ) para comprobar si existía asociación entre el empeoramiento clínico que requiriera cirugía y: 1-la edad del paciente; 2-diabetes; 3- un patrón de diabetes en la familia próxima; y 4-enfermedad de Dupuytren. Resultados: En relación con la curvatura y el número y tamaño de las placas fibrosas se ha observado una tendencia hacia la estabilización del grupo de pacientes de más de 50 años de edad. El 68% de los pacientes del grupo de edad menor de 50 años mostraron un empeoramiento progresivo de la enfermedad requiriendo cirugía, mientras que en el otro grupo sólo el 31,5% de los pacientes necesitaron cirugía. El análisis estadístico confirmó que el empeoramiento de la enfermedad es más probable en el grupo menor de 50 años de edad (OR= 3,5, IC 2-8) y en los pacientes diabéticos (ajustando por edad OR = 6, IC 2-19). El análisis estadístico no ha mostrado diferencias significativas en la evolución de la enfermedad de Peyronie en cuanto a la presencia de un patrón de diabetes en los familiares cercanos o de enfermedad de Dupuytren. Conclusiones: Los pacientes presentaron una discreta tendencia hacia la estabilización espontánea. Los pacientes diagnosticados antes de los 50 tiene una mayor probabilidad de empeoramiento de la enfermedad requiriendo un abordaje quirúrgico. Los datos sobre la historia natural de la enfermedad de Peyronie en pacientes no tratados deben transmitir cautela a la hora de considerar la eficacia de los nuevos tratamientos farmacológicos y físicos (AU)