Homogeneity and persistence of transgene expression by omitting antibiotic selection in cell line isolation

Nonuniform, mosaic expression patterns of transgenes are often linked to transcriptional silencing, triggered by epigenetic modifications of the exogenous DNA. Such phenotypes are common phenomena in genetically engineered cells and organisms. They are widely attributed to features of transgenic transcription units distinct from endogenous genes, rendering them particularly susceptible to epigenetic downregulation. Contrary to this assumption we show that the method used for the isolation of stably transfected cells has the most profound impact on transgene expression patterns. Standard antibiotic selection was directly compared to cell sorting for the establishment of stable cells. Only the latter procedure could warrant a high degree of uniformity and stability in gene expression. Marker genes useful for the essential cell sorting step encode mostly fluorescent proteins. However, by combining this approach with site-specific recombination, it can be applied to isolate stable cell lines with the desired expression characteristics for any gene of interest

Therapeutic Hypothermia after Peri-Interventional In-Hospital Cardiac Arrest

Background: Therapeutic hypothermia is recommended by international guidelines for patients after out-of-hospital cardiac arrest (OHCA) due to ventricular fibrillation. However, data for patients after in-hospital cardiac arrest (IHCA) are still scarce. Guidelines leave it to the attending physician to decide on the use of hypothermia in IHCA patients.Objective: To determine the use of therapeutic hypothermia in-hospital cardiac arrest.Design: Retrospective case series.Setting: University Hospital of colgne, intensive care units.Subjects: Seven patients admitted to the intensive care unit after peri-interventional IHCA between January and December 2009.Interventions: Therapeutic hypothermia was initiated in all patients with a median delay of five hours.Results: Four out of seven patients (57 %) survived cardiac arrest, but one of these later died due to her primary cause of hospitalisation. The other three patients were discharged without neurological sequelae. There were no serious adverse effects of therapeutic hypothermia.Conclusion: Therapeutic hypothermia after peri-interventional IHCA IHCA is safe and might benefit the patient. This treatment strategy should be taken into consideration until further data are available

Anesthesiological management of Caesarean sections. Nationwide survey in Germany

Background. The rate of Caesarean sections in Germany continues to rise. The change in anesthetic technique of choice from general to spinal anesthesia began later than in other countries and at the last survey in 2002 was not widely established. The literature on the anesthetic management of Caesarean sections contains many controversies, for example fluid preload before performing spinal anesthesia and the vasopressor of choice. Other issues have received relatively little attention, such as the level of experience of anesthesiologists working autonomously on the labour ward or the timing of antibiotic prophylaxis. The aim of the current survey was to provide an updated overview of anesthetic management of Caesarean sections in Germany. Material and Methods. A questionnaire was sent out to 709 departments of anesthesiology serving obstetric units in Germany. The questionnaire concerned various aspects of anesthetic management of Caesarean sections. Results. A total of 360 questionnaires (50.8%) were returned of which 346 were complete and could be analyzed, accounting for 330,000 births and 90,000 Caesarean sections per year. The predominant anesthetic method used for Caesarean sections was spinal anesthesia (90.8%) using hyperbaric bupivacaine and in approximately one third of the hospitals surveyed without administering intrathecal opioids. Approximately 12% of the departments surveyed used traumatic Quincke needles. In 86.2% the vasopressor of choice was caffedrine/theodrenaline. Nitrous oxide was used in only 19.2% of departments surveyed when general anesthesia is performed. An antibiotic drug was administered in only 11% of hospitals before cord clamping. In 43.1% no neonatologist was available to treat unexpected critically ill newborns. In 32.1% of departments surveyed residents with less than 2 years experience worked autonomously on the labour ward. Conclusions. Currently the predominant anesthetic technique of choice in Germany is spinal anaesthesia and at a much higher rate than in 2002. In addition 12% of departments use traumatic Quincke needles which are associated with a higher incidence of postpuncture headache. Nitrous oxide is no longer frequently used in Germany. Finally, the administration of an antibiotic before cord clamping has been shown to lead to lower rates of endometritis and postoperative wound infection without detrimental effects on the newborn. This is practiced in only a small minority of departments across Germany