Optimization of mRNA extraction from human nasal mucosa biopsies for gene expression profile analysis by qRT-PCR.

Quantitative real-time reverse transcriptase polymerase chain reaction (qRT-PCR) is the gold-standard method for analyzing modifications in gene expression in cells and tissues. However, large quantities of high-quality RNA samples are needed for analyzing the expression of multiple genes from one human tissue sample. Here, we provide an optimized protocol for extracting large amounts of RNA from human nasal mucosal biopsies. The quality and quantity of samples were sufficient for qRT-PCR analyses of the expressions of various genes, in duplicate. In contrast to other protocols, we optimized RNA isolation to increase the amount from nasal biopsy samples for analyses of multiple genes. In most previous publications, expressions of only one or a few genes, including housekeeping genes, were analyzed because the amount of biological material was small. We were able to improve our protocol with respect to the yield and quality of RNA. This is likely to produce better results from molecular analyses of very small biopsy samples of human nasal mucosa.

Palaeoecological data indicates land-use changes across Europe linked to spatial heterogeneity in mortality during the Black Death pandemic.

The Black Death (1347-1352 CE) is the most renowned pandemic in human history, believed by many to have killed half of Europe's population. However, despite advances in ancient DNA research that conclusively identified the pandemic's causative agent (bacterium Yersinia pestis), our knowledge of the Black Death remains limited, based primarily on qualitative remarks in medieval written sources available for some areas of Western Europe. Here, we remedy this situation by applying a pioneering new approach, 'big data palaeoecology', which, starting from palynological data, evaluates the scale of the Black Death's mortality on a regional scale across Europe. We collected pollen data on landscape change from 261 radiocarbon-dated coring sites (lakes and wetlands) located across 19 modern-day European countries. We used two independent methods of analysis to evaluate whether the changes we see in the landscape at the time of the Black Death agree with the hypothesis that a large portion of the population, upwards of half, died within a few years in the 21 historical regions we studied. While we can confirm that the Black Death had a devastating impact in some regions, we found that it had negligible or no impact in others. These inter-regional differences in the Black Death's mortality across Europe demonstrate the significance of cultural, ecological, economic, societal and climatic factors that mediated the dissemination and impact of the disease. The complex interplay of these factors, along with the historical ecology of plague, should be a focus of future research on historical pandemics.

Novel functions of S1P in chronic itchy and inflammatory skin diseases.

S1P is a pleotropic sphingolipid signalling molecule that acts through binding to five high-affinity G-protein coupled receptors. S1P-signaling affects cell fate in a multitude of ways, e.g. influencing cell differentiation, proliferation, and apoptosis, as well as playing an important role in immune cell trafficking. Though many effects of S1P-signaling in the human body have been discovered, the full range of functions is yet to be understood. For inflammatory skin diseases such as atopic dermatitis and psoriasis, evidence is emerging that dysfunction and imbalance of the S1P-axis is a contributing factor. Multiple studies investigating the efficacy of S1PR modulators in alleviating the severity and symptoms of skin conditions in various animal models and human clinical trials have shown promising results and validated the interest in the S1P-axis as a potential therapeutic target. Even though the involvement of S1P-signalling in inflammatory skin diseases still requires further clarification, the implications of the recent findings may prompt expansion of research to additional skin conditions and more S1P-axis modulatory pharmaceuticals.

Aluminium release and fluid warming: provocational setting and devices at risk.

BACKGROUND: Fluid warming, recommended for fluid rates of > 500 ml h-1, is an integral part of patient temperature management strategies. Fluid warming devices using an uncoated aluminium containing heating element have been reported to liberate aluminium resulting in critical aluminium concentrations in heated fluids. We investigated saline solution (0.9%), artificially spiked with organic acids to determine the influence of fluid composition on aluminium release using the uncoated enFlow® device. Additionally, the Level1® as a high volume fluid warming device and the ThermoSens® device were investigated with artificial spiked fluid at high risk for aluminum release and a clinically used crystalloid solution. RESULTS: Saline solution spiked with lactate more than acetate, especially at a non neutral pH, led to high aluminium release. Next to the enFlow® device, aluminium release was observed for the Level1® device, but not for the coated ThermoSens®-device. CONCLUSION: Uncoated aluminium containing fluid warming devices lead to potentially toxic levels of aluminium in heated fluids, especially in fluids with non-neutral pH containing organic acids and their salts like balanced electrolyte solutions.

Breakthrough in purification of fossil pollen for dating of sediments by a new large-particle on-chip sorter.

Particle sorting is a fundamental method in various fields of medical and biological research. However, existing sorting applications are not capable for high-throughput sorting of large-size (>100 micrometers) particles. Here, we present a novel on-chip sorting method using traveling vortices generated by on-demand microjet flows, which locally exceed laminar flow condition, allowing for high-throughput sorting (5 kilohertz) with a record-wide sorting area of 520 micrometers. Using an activation system based on fluorescence detection, the method successfully sorted 160-micrometer microbeads and purified fossil pollen (maximum dimension around 170 micrometers) from lake sediments. Radiocarbon dates of sorting-derived fossil pollen concentrates proved accurate, demonstrating the method's ability to enhance building chronologies for paleoenvironmental records from sedimentary archives. The method is capable to cover urgent needs for high-throughput large-particle sorting in genomics, metabolomics, and regenerative medicine and opens up new opportunities for the use of pollen and other microfossils in geochronology, paleoecology, and paleoclimatology.

Conductive heating mattress leads to ECG changes that mimic pacemaker spikes.

Hypothermia is a common perioperative complication. To prevent perioperative hypothermia amongst other things electrical heating mattresses are used. We have made an observation with the use of an electrical heating mattress that may confuse users. In this case the ECG monitoring suddenly showed spikes that looked like spikes from an implanted pacemaker. When turning off the heating mattress the spikes disappeared and returned after turning on the heating mattress again.

[Intraoperative vascular air embolism : Evidence for risks, diagnostics and treatment]. / Intraoperative vaskuläre Luftembolie : Evidenz bei Risiko, Diagnostik und Therapie.

The entry of gases into the vascular system is called vascular air embolism (VAE). The blocking of the pulmonary circulation by VAE can lead to fulminant right-sided heart failure and cardiocirculatory arrest. A VAE can occur at any time if there is an open connection between the environment and a venous vessel with subatmospheric pressure. This situation occurs during ear nose throat surgery, hip surgery, surgery of the lesser pelvis or breast surgery, if the surgical field is above the level of the heart; however, a VAE can also occur during routine tasks, such as insertion or removal of a central venous catheter or during endoscopic procedures with the insufflation of gas.Because during these procedures VAE is not the main focus of the anesthesia or surgery personnel, in such situations its sudden unexpected occurrence can have severe consequences. In contrast, in cardiac surgery or neurosurgery the risk of intraoperative VAE is much better known. In procedures with a higher risk of a clinically relevant VAE, a patent foramen ovale should be ruled out by preoperative transesophageal echocardiography (TEE). Intraoperatively TEE is the most sensitive procedure not only to detect a VAE but also to visualize the clinical expression, e.g. acute right heart overload.The avoidance of an initial and repeated air embolism is the primary measure to minimize the incidence and severity of VAE.Intraoperatively the following measures should be undertaken: excellent communication between anesthesia and surgery personnel with predetermined actions, maintenance of normal volume, patient positioning with minimal difference in height between heart and head, state of the art surgical technique with closure of potential air entry sites, sufficient detection of air by TEE, repeated jugular vein compression during neurosurgery, intraoperative Trendelenburg positioning of the patient during persisting or clinically evident VAE, differentiated adjustment of ventilatory settings and catecholamine treatment, aspiration of the blood-air mixture (air lock) at the junction of the superior vena cava and right atrium through a large bore central venous line and keeping check of the coagulation status.

Aluminium release by coated and uncoated fluid-warming devices.

The use of fluid-warming systems is recommended for infusion rates > 500 ml.h-1 to avoid peri-operative hypothermia. Some fluid-warming devices use disposable aluminium-heated plates for heat transfer, but there is no protective coating to separate the fluid from the heated aluminium surface. It is unknown if this could promote release of aluminium into infusion fluids. We investigated a coated (Fluido compact) and an uncoated (enFlow) fluid-warming device using normal saline or balanced electrolyte solution as infusion fluids, pumped through the heated disposables at flow rates of 2, 4 and 8 ml.min-1 for 60 min each. Aluminium concentrations in the fluid samples were analysed using graphite furnace atomic absorption spectrometry. With saline the coated and uncoated devices yielded aluminium concentrations below the level of quantification (< 128 µg.l-1 ). Similarly, balanced electrolyte solution in the coated device yielded aluminium concentrations < 128 µg.l-1 . However, balanced electrolyte solution in the uncoated device yielded aluminium concentrations of up to 6794 (3465-8002 [1868-7421]) µg.l-1 . Repeating this last study at a flow rate of 2 ml.min-1 resulted in quite high aluminium concentrations when the uncoated device was not heated (~1000 µg.l-1 ) and higher concentrations after the device was heated. We conclude that using uncoated aluminium plates in fluid-warming systems can lead to a risk of administering potentially harmful concentrations of aluminium when balanced crystalloid solutions are used. The mechanism is unclear, but heat is in part involved. Coating for aluminium within medical devices in direct contact with infusion fluids should be recommended.

[Prewarming according to the AWMF S3 guidelines on preventing inadvertant perioperative hypothermia 2014 : Retrospective analysis of 7786 patients]. / Vorwärmung entsprechend der "S3 Leitlinie Vermeidung von unbeabsichtigter perioperativer Hypothermie 2014" : Retrospektive Analyse von 7786 Patienten.

BACKGROUND: Inadvertent perioperative hypothermia, which is defined as a core body temperature of less than 36.0 °C, can have serious consequences in surgery patients. These include cardiac complications, increased blood loss, wound infections and postoperative shivering; therefore, the scientific evidence that inadvertent perioperative hypothermia should be avoided is undisputed and several national guidelines have been published summarizing the scientific evidence and recommending specific procedures. The German AWMF guidelines were the first to emphasize the importance of prewarming for surgery patients to avoid inadvertant perioperative hypothermia; however, in contrast to intraoperative warming, prewarming is so far not sufficiently implemented in clinical practice in many hospitals. Furthermore, a recent study has questioned the effectiveness of prewarming. OBJECTIVE: The aim of this retrospective investigation was to evaluate the hypothermia rates that can be achieved when prewarming in the anesthesia induction room is introduced into the clinical practice and performed in addition to intraoperative warming. MATERIAL AND METHODS: The ethics committee of the Medical Faculty of the Martin Luther University Halle Wittenberg gave approval for data storage and retrospective data analysis from the anesthesia database. According to the existing local standard operating procedure, prewarming with forced air was performed in addition to intraoperative warming in the anesthesia induction room in 3899 patients receiving general anesthesia with a duration of 30 min or longer from January 2015 to December 2016. The results were compared with a control group of 3887 patients from July 2012 to August 2014 who received intraoperative warming but were not subjected to prewarming. Tracheal intubation was carried out in all patients and temperature measurements after the induction of anesthesia were performed using esophageal, urinary catheter or intra-arterial temperature probes. RESULTS: The mean duration of prewarming was 25 min in the treatment group. Patients subjected to prewarming showed an intraoperative hypothermia rate of 15.8% and a postoperative hypothermia rate of 5.1%. Patients without prewarming showed an intraoperative hypothermia rate of 30.4% and a postoperative hypothermia rate of 12.4%. This means a 52% reduction of the intraoperative hypothermia rate and a 41% reduction of the postoperative hypothermia rate for patients who received prewarmimg (p < 0.0001). Multivariate logistic regression revealed that the lack of prewarming was independently associated with intraoperative hypothermia with an odds ratio of 2.5 (95% confidence interval CI 2.250-2.841; p < 0.0001) and postoperative hypothermia with an odds ratio of 2.8 (95% CI 2.316-3.277; p < 0.0001). CONCLUSION: Prewarming, as recommended in the AWMF guidelines, resulted in a significant and clinically relevant reduction in the incidence of inadvertent perioperative hypothermia; therefore, prewarming can still be regarded as an effective method to avoid perioperative hypothermia. Hypothermia rates of 15.8% intraoperatively and 5.1% postoperatively can be achieved in clinical practice, when prewarming is performed in addition to intraoperative warming in the anesthesia induction room directly before the start of surgical procedures.

[Neurophysiological monitoring during surgical procedures]. / Neurophysiologisches Monitoring bei operativen Eingriffen.

The application of intraoperative neurophysiological monitoring (IONM) is gaining more and more importance in daily clinical practice. The use of IONM allows the localization of neural structures and to control functioning of the peripheral and central nervous systems in anesthetized patients. This enables surgeons to identify and to protect neural structures and cerebral areas. The use of IONM also enables anesthesiologists to adjust anesthesia and cardiopulmonary therapy to the individual needs of the patient. Thereby, it may be possible to reduce the incidence of postoperative delirium and the incidence of postoperative cognitive deficits. To exploit the full potential anesthesiologists and surgeons must be able to use the methods of IONM safely and understand the results; therefore, basic knowledge of the technology, options and limitations of IONM is necessary. It is also important to be aware of the influence of anesthetics on the methods of IONM. Total intravenous anesthesia (TIVA) is the anesthetic method of choice, because it has only minimal influence on IONM methods. It is important to avoid bolus injections of hypnotics to achieve stable blood concentrations. Long- acting neuromuscular blocking agents should be avoided, because they disturb the signals of electromyography and motor-evoked potentials. By using IONM anesthesiologists and surgeons can identify changes in the function of the peripheral and central nervous system prior to irreversible damage.

Plasmonic Purcell effect reveals obliquely ordered phosphorescent emitters in Organic LEDs.

The non-isotropic alignment of molecules can increase the interaction efficiency with propagating light fields. This applies to both emissive and absorptive systems and can be exploited for achieving unprecedented efficiencies of organic opto-electronic devices such as organic light-emitting diodes. Optical analysis has revealed certain phosphorescent emitters to align spontaneously in an advantageous orientation. Unfortunately, established approaches only determine an average orientation because emission patterns solely depend on the second moments of the transition dipole vector distribution. In order to resolve further details of such a distribution, additional differences in the emission characteristics of parallel and perpendicularly oriented emitters need to be introduced. A thin metal layer near the emitters introduces plasmon mediated losses mostly for perpendicular emitters. Then, analyzing the emission at different polarizations allows one to measure emission lifetimes of mostly parallel or mostly perpendicular oriented emitters. This should alter the transient emission when observing the temporal phosphorescence decay under different directions and/or polarizations. The angular width of the orientation distribution can be derived from the degree of such lifetime splitting. Our results suggest a narrow but obliquely oriented molecular ensemble of Ir(MDQ)2(acac) doped into the α-NPD host inside an Organic LED stack.

Haptoglobin and free haemoglobin during cardiac surgery-is there a link to acute kidney injury?

Acute kidney injury (AKI) is frequently observed after cardiac surgery (CS) with cardiopulmonary bypass (CPB). Multiple mechanisms underlie this phenomenon, including CPB-dependent haemolysis. Haemoglobin is released during haemolysis, and free haemoglobin (frHb) causes tubular cell injury after exceeding the binding capacity of haptoglobin (Hp). The objective of this study was to investigate the influence of perioperative changes in frHb and Hp levels on the incidence of CS-associated (CSA) AKI. After receiving local ethics committee approval and obtaining informed consent from our patients, we analysed the data pertaining to 154 patients undergoing CPB surgery. We recorded frHb and Hp concentrations pre-, intra- and postoperatively and defined AKI using the Kidney Disease Improving Global Outcomes (KDIGO) classification. We observed that frHb levels increased significantly during surgery and then decreased at ten hours thereafter and that Hp levels decreased during surgery and remained at low levels until the first postoperative day. We noted a moderate negative correlation between frHb and Hp levels. AKI was identified in 45.5% of patients; however, there was no significant difference in frHb or Hp levels between patients with and without AKI. We did not observe a relationship between frHb or Hp levels and CSA AKI and thus could not confirm the hypothesis that patients with higher baseline Hp concentrations experience a lower incidence of AKI than patients with lower baseline Hp concentrations.

[Prevention of perioperative hypothermia : Implementation of the S3 guideline]. / Vermeidung perioperativer Hypothermie : Umsetzung der S3-Leitlinie.

To improve perioperative quality and patient safety, the German S3 guideline should be consistently implemented to avoid perioperative hypothermia. Perioperative normothermia is a quality indicator and should be achieved by anesthesiologists and surgeons. To detect hypothermia early during the perioperative process, measuring body temperature should be started 1-2 h preoperatively. Patients should be actively warmed for 20-30 min before starting anesthesia. Prewarming is most effective and should be included in the preoperative process. Patients should be informed about the risks of perioperative hypothermia and members of the perioperative team should be educated. A standard operating procedure (SOP) to avoid hypothermia should be introduced in every operative unit. The incidence of postoperative hypothermia should be evaluated in operative patients every 3-6 months. The goals should be to measure body temperature in >80% of patients undergoing surgery and for >70% to exhibit a core temperature >36 °C at the end of surgery.

Three millennia of heavy rainfalls in Western Mediterranean: frequency, seasonality and atmospheric drivers.

Documenting subdecadal-scale heavy rainfall (HR) variability over several millennia can rarely be accomplished due to the paucity of high resolution, homogeneous and continuous proxy records. Here, using a unique, seasonally resolved lake record from southern Europe, we quantify temporal changes in extreme HR events for the last 2,800 years in this region and their correlation with negative phases of the Mediterranean Oscillation (MO). Notably, scarce HR dominated by a persistent positive MO mode characterizes the so-called Migration period (CE 370-670). Large hydroclimatic variability, particularly between CE 1012 and 1164, singles out the Medieval Climatic Anomaly, whereas more stationary HR conditions occurred between CE 1537 and 1805 coinciding with the Little Ice Age. This exceptional paleohydrological record highlights that the present-day trend towards strengthened hydrological deficit and less HR in the western Mediterranean is neither acute nor unusual in the context of Late Holocene hydrometeorological variability at centennial to decadal time scales.

The effects of residual pump blood on patient plasma free haemoglobin levels post cardiac surgery.

At the end of cardiopulmonary bypass, there are invariably several hundred millilitres of residual pump blood in the reservoir, which can either be re-transfused or discarded. The objective of this prospective observational study was to investigate the quality of the residual pump blood, focusing on plasma free haemoglobin (pfHb) and blood cell counts. Fifty-one consecutive patients were included in the study. Forty-nine units of residual pump blood and 58 units of transfused red blood cell (RBC) concentrates were analysed. The mean preoperative pfHb of the patients was 0.057 ± 0.062 g/l, which increased gradually to 0.55 ± 0.36 g/l on arrival in the intensive care unit postoperatively. On the first postoperative day, the mean pfHb had returned to within the normal range. Our data showed that haemoglobin, haematocrit, and erythrocyte counts of residual pump blood were approximately 40% of the values in standardised RBC concentrates. Plasma free haemoglobin was significantly higher in residual pump blood compared to RBC concentrates, and nearly twice as high as the pfHb in patient blood samples taken contemporaneously. Our findings indicate that residual pump blood pfHb levels are markedly higher compared to patients' blood and RBC concentrates, but that its administration does not significantly increase patients' pfHb levels.

[Tracheal bronchus and contralateral pneumonectomy]. / Trachealer Bronchusabgang und kontralaterale Pneumonektomie.

One-lung ventilation is a standard procedure for many types of lung surgery. The anesthesiologist can be challenged if unknown anomalies of the bronchial tree occur. We report a patient with a tracheal bronchus on the right side presenting for left pneumonectomy, and present one possible solution to airway management.

[Implementation of a thermal management concept to prevent perioperative hypothermia : Results of a 6­month period in clinical practice]. / Umsetzung eines Wärmemanagementkonzeptes zur Vermeidung von perioperativer Hypothermie : Ergebnisse eines Sechsmonatszeitraums in der klinischen Praxis.

BACKGROUND: Perioperative hypothermia is defined as a core temperature below 36 °C. The literature shows that perioperative hypothermia is a frequent but potentially preventable complication of the surgical process. The risk of experiencing perioperative hypothermia is inherent for all anesthetized patients, independent of the type of surgery. Unless preventative measures are taken, perioperative hypothermia occurs in 50 to 70 % of all surgical patients. In Germany and Austria the guideline "Preventing inadvertent perioperative hypothermia" has been published. In Wolfsburg we started already in 2012 with a standard operating procedure to prevent perioperative hypothermia in all surgical patients. In two clinical departments we established an additional prewarming-protocol starting prior to induction of anaesthesia on the normal ward on the day of surgery. MATERIAL AND METHODS: For a period of 6 months we analyzed all temperature data of patients having undergone surgery, beginning before the start of general anaesthesia until the end of the operation. RESULTS: In total 3228 patients were enrolled into the study. Prewarming was performed in 1329 patients. In 1902 patients active warming was limited to the intraoperative period. The total rate of hypothermia in all patients was 32.6 %, whereas the rate of hypothermia at the end of the operation was 19.3 %. In the group of patients without prewarming the overall rate was 39.1 vs. 25 % at the end of the operation. In the groups of patients with prewarming the total rates of hypothermia were 25.2 and 24.7 % overall and 14.4 and 12.5 % at the end of the operation. In multifactorial regression it could be shown that patients without prewarming had a 1.8-fold increased risk of perioperative hypothermia compared to patients with intraoperative warming only. CONCLUSION: We conclude that temperature management is a challenge in the clinical situation, and that it is difficult to achieve rates of hypothermia close to zero. The addition of prewarming was very effective in improving the results in our patients.

Osteogenic differentiation of immature osteoblasts: Interplay of cell culture media and supplements.

Differentiation of immature osteoblasts to mature osteoblasts in vitro initially was induced by supplementing the medium with ß-gylcerophosphate and dexamethasone. Later, ascorbic acid, vitamin D3, vitamin K3 and TGFß1 were used in varying concentrations as supplements to generate a mature osteoblast phenotype. We tested the effects of several combinations of cell culture media, seeding protocols and osteogenic supplements on osteogenic differentiation of human primary osteoblasts. Osteogenic differentiation was analyzed by staining alkaline phosphatase (ALP) with 5-bromo-4-chloro-3-indolyl-phosphate/nitro blue tetrazolium (BCIP/NBT) and by von Kossa staining of deposited calcium phosphate. The combinations of culture media and supplements significantly influenced osteogenic differentiation, but the seeding protocol did not. Staining of ALP and calcium phosphate could be achieved only if our own mix of osteogenic supplements was used in combination with Dulbecco's modified Eagle medium or if a commercial mix of osteogenic supplements was used in combination with osteoblast growth medium. Especially for von Kossa, we observed great variations in the staining intensity. Because osteogenic differentiation is a complex process, the origin of the osteoblasts, cell culture media and osteogenic supplements should be established by preliminary experiments to achieve optimal differentiation. Staining of ALP or deposited calcium phosphate should be supplemented with qRT-PCR studies to learn more about the influence of specific supplements on osteogenic markers.

[Perioperative thermal management in Germany varies depending on the hospital size]. / Perioperatives Wärmemanagement in Abhängigkeit von der Krankenhausgröße in Deutschland.

BACKGROUND: Improvement of quality of care and patient safety while decreasing costs are major challenges in healthcare systems. This challenge includes the avoidance of perioperative hypothermia to reduce the associated adverse effects, length of stay and treatment costs. Due to the medical and economic relevance the national S3 guidelines for the prevention of perioperative hypothermia were recently published. AIM: This study presents and analyses the reality of utilization of thermal management in German hospitals depending on the size of the hospital, which is based on the number of beds. MATERIAL AND METHODS: Based on the data of an online survey among all members of the German Society of Anesthesiology and Intensive Care Medicine about perioperative thermal management, a subgroup analysis differentiating between the size of hospitals was performed. The survey included questions about the structural and organizational conditions, the practical implementation of temperature measurement and warming therapy and the developmental status of clinical standard operating procedures (SOP) and educational training. RESULTS: Comparing the structural quality, major differences were found with respect to the availability of core body temperature measurement and the provision of warming devices especially at different peripheral anesthesia workplaces as well as the existence of SOPs and educational training. The availability increased with hospital size. With respect to process quality, the frequency of prewarming increased with hospital size as well as the frequency of intraoperative temperature measurements during different anesthesia procedures. CONCLUSION: Major differences were found in several aspects of perioperative thermal management depending on the hospital size. The main potential for improvement was found in smaller hospitals. Developmental needs primarily exist in the configuration of peripheral anesthesia workplaces, educational training, implementation of SOPs and prewarming of patients.