Экономические перспективы повышения уровня использования попутного нефтяного газа

A novel method employing filter arrays of a cDNA expression library for the identification of substrates for protein kinases was developed. With this technique, we identified a new member of the cyclin family, cyclin L2, as a substrate of the nuclear protein kinase DYRK1A. Cyclin L2 contains an N-terminal cyclin domain and a C-terminal arginine/serine-rich domain (RS domain), which is a hallmark of many proteins involved in pre-mRNA processing. The gene for cyclin L2 encodes the full-length cyclin L2, which is predominantly expressed in testis, as well as a truncated splicing variant (cyclin L2S) that lacks the RS domain and is ubiquitously expressed in human tissues. Full-length cyclin L2, but not cyclin L2S, was associated with the cyclin-dependent kinase PITSLRE. Cyclin L2 interacted with splicing factor 2 in vitro and was co-localized with the splicing factor SC35 in the nuclear speckle compartment. Photobleaching experiments showed that a fusion protein of green fluorescent protein and cyclin L2 in nuclear speckles rapidly exchanged with unbleached molecules in the nucleus, similar to other RS domain-containing proteins. In striking contrast, the closely related green fluorescent protein-cyclin L1 was immobile in the speckle compartment. DYRK1A interacted with cyclin L2 in pull-down assays, and overexpression of DYRK1A stimulated phosphorylation of cyclin L2 in COS-7 cells. These data characterize cyclin L2 as a highly mobile component of nuclear speckles and suggest that DYRK1A may regulate splicing by phosphorylation of cyclin L2

Hyperoxia-Induced Protein Alterations in Renal Rat Tissue: A Quantitative Proteomic Approach to Identify Hyperoxia-Induced Effects in Cellular Signaling Pathways

Introduction. In renal tissue as well as in other organs, supranormal oxygen pressure may lead to deleterious consequences on a cellular level. Additionally, hyperoxia-induced effect in cells and related free radicals may potentially contribute to renal failure. The aim of this study was to analyze time-dependent alterations of rat kidney protein expression after short-term normobaric hyperoxia using proteomics and bioinformatic approaches. Material and Methods. N=36 Wistar rats were randomized into six different groups: three groups with normobaric hyperoxia (exposure to 100% oxygen for 3 h) and three groups with normobaric normoxia (NN; room air). After hyperoxia exposure, kidneys were removed immediately, after 3 days and after 7 days. Kidney lysates were analyzed by two-dimensional gel electrophoresis followed by peptide mass fingerprinting using tandem mass spectrometry. Statistical analysis was performed with DeCyder 2D software (p<0.01). Biological functions of differential regulated proteins were studied using functional network analysis (Ingenuity Pathways Analysis and PathwayStudio). Results. Expression of 14 proteins was significantly altered (p<0.01): eight proteins (MEP1A_RAT, RSSA_RAT, F16P1_RAT, STML2_RAT, BPNT1_RAT, LGMN_RAT, ATPA_RAT, and VDAC1_RAT) were downregulated and six proteins (MTUS1_RAT, F16P1_RAT, ACTG_RAT, ACTB_RAT, 2ABA_RAT, and RAB1A_RAT) were upregulated. Bioinformatic analyses revealed an association of regulated proteins with inflammation. Conclusions. Significant alterations in renal protein expression could be demonstrated for up to 7 days even after short-term hyperoxia. The identified proteins indicate an association with inflammation signaling cascades. MEP1A and VDAC1 could be promising candidates to identify hyperoxic injury in kidney cells

Charakterisierung von Cyclin L2 : ein neuer Vertreter der Cyclin-Familie

The dual-specifity tyrosin (Y) phosphorylation-regulated kinase-1A (DYRK1A) plays a prominent role in development of the central nervous system. The gene is located within the down-syndrome-critical-region on chromosome 21. The molecular mediation of the protein function of DYRK1A remains unclear. Cyclin L2, a recently identified substrates of DYRK1A, combines the unique combination of cyclin domain and arginine-serine-rich domain (rs-domain). These rs-domains comprise a hallmark for splicing factors. The intention of this thesis was to characterize cyclin L2 and to analyze the interaction between DYRK1A and cyclin L2. Application of a cyclin L2-specific antibody revealed in vivo expression in isolated nuclei of mouse-testes. Subcellular localization of cyclin L2 was assessed via live-cell-imaging. Cyclin L2, and the closely related cyclin L1, are organized in nuclear speckles, known to be the place of supply, modification and storage of splicing factors. The fact that DYRK1A is allocated in nuclear speckles as well indicates a physiological interaction between both proteins. Furthermore this thesis demonstrates that overexpressed DYRK1A but not the catalytic inactive form of DYRK1A generates an additional band of cyclin L2 with less electrophoretic mobility in western blot assays, which is typical for phosphorylation of proteins and refers to a phosphorylation of cyclin L2 by DYRK1A. Western blot assays also prove the interaction of cyclin L2 and the endogenous cyclin dependent kinase PITSLRE p110 (cdk11) in vivo. Compared to cyclin L1 and PITSLREp110 this interaction is weaker and so far no other catalytic active partner of cyclin L2 is known. In contrast to this analogy of both L-type-cyclins photobleaching-technique visualized that cyclin L1 and cyclin L2 act entirely opposed: cyclin L2 proved to be highly mobile within the nucleus whereas cyclin L1 seemed to be totally immobile, a unique and characteristic trait for proteins with an rs-domain. By using chimeric proteins the rs-domain was held to be responsible for this opposed mobility behavior, which was as yet an undescribed function of rs-domains. This thesis characterizes cyclin L2 as a new member of the cyclin family with specific properties different from cyclin L1. The results assume an interaction between cyclin L2 and DYRK1A in vivo and therefore DYRK1A might play a role concerning posttranscriptional modification by splicing

Charakterisierung von Cyclin L2 : ein neuer Vertreter der Cyclin-Familie

The dual-specifity tyrosin (Y) phosphorylation-regulated kinase-1A (DYRK1A) plays a prominent role in development of the central nervous system. The gene is located within the down-syndrome-critical-region on chromosome 21. The molecular mediation of the protein function of DYRK1A remains unclear. Cyclin L2, a recently identified substrates of DYRK1A, combines the unique combination of cyclin domain and arginine-serine-rich domain (rs-domain). These rs-domains comprise a hallmark for splicing factors. The intention of this thesis was to characterize cyclin L2 and to analyze the interaction between DYRK1A and cyclin L2. Application of a cyclin L2-specific antibody revealed in vivo expression in isolated nuclei of mouse-testes. Subcellular localization of cyclin L2 was assessed via live-cell-imaging. Cyclin L2, and the closely related cyclin L1, are organized in nuclear speckles, known to be the place of supply, modification and storage of splicing factors. The fact that DYRK1A is allocated in nuclear speckles as well indicates a physiological interaction between both proteins. Furthermore this thesis demonstrates that overexpressed DYRK1A but not the catalytic inactive form of DYRK1A generates an additional band of cyclin L2 with less electrophoretic mobility in western blot assays, which is typical for phosphorylation of proteins and refers to a phosphorylation of cyclin L2 by DYRK1A. Western blot assays also prove the interaction of cyclin L2 and the endogenous cyclin dependent kinase PITSLRE p110 (cdk11) in vivo. Compared to cyclin L1 and PITSLREp110 this interaction is weaker and so far no other catalytic active partner of cyclin L2 is known. In contrast to this analogy of both L-type-cyclins photobleaching-technique visualized that cyclin L1 and cyclin L2 act entirely opposed: cyclin L2 proved to be highly mobile within the nucleus whereas cyclin L1 seemed to be totally immobile, a unique and characteristic trait for proteins with an rs-domain. By using chimeric proteins the rs-domain was held to be responsible for this opposed mobility behavior, which was as yet an undescribed function of rs-domains. This thesis characterizes cyclin L2 as a new member of the cyclin family with specific properties different from cyclin L1. The results assume an interaction between cyclin L2 and DYRK1A in vivo and therefore DYRK1A might play a role concerning posttranscriptional modification by splicing

Response of rat lung tissue to short-term hyperoxia: a proteomic approach

An inspiratory oxygen fraction of 1.0 is often required to avoid hypoxia both in many pre- and in-hospital situations. On the other hand, hyperoxia may lead to deleterious consequences (cell growth inhibition, inflammation, and apoptosis) for numerous tissues including the lung. Whereas clinical effects of hyperoxic lung injury are well known, its impact on the expression of lung proteins has not yet been evaluated sufficiently. The aim of this study was to analyze time-dependent alterations of protein expression in rat lung tissue after short-term normobaric hyperoxia (NH). After approval of the local ethics committee for animal research, N = 36 Wistar rats were randomized into six different groups: three groups with NH with exposure to 100 % oxygen for 3 h and three groups with normobaric normoxia (NN) with exposure to room air (21 % oxygen). After the end of the experiments, lungs were removed immediately (NH0 and NN0), after 3 days (NH3 and NN3) and after 7 days (NH7 and NN7). Lung lysates were analyzed by two-dimensional gel electrophoresis (2D-GE) followed by peptide mass fingerprinting using mass spectrometry. Statistical analysis was performed with Delta 2D (DECODON GmbH, Greifswald, Germany; ANOVA, Bonferroni correction, p < 0.01). Biological functions of differential regulated proteins were studied using functional network analysis (Ingenuity Pathways Analysis, IPA). pO(2) was significantly higher in NH-groups compared to NN-groups (581 +/- A 28 vs. 98 +/- A 12 mmHg; p < 0.01), all other physiological parameters did not differ. Expression of 14 proteins were significantly altered: two proteins were up-regulated and 12 proteins were down-regulated. Even though NH was comparatively short termed, significant alterations in lung protein expression could be demonstrated up to 7 days after hyperoxia. The identified proteins indicate an association with cell growth inhibition, regulation of apoptosis, and approval of structural cell integrity

APPLICATION OF THE FIA SCORE TO GERMAN RESCUE HELICOPTER ACCIDENTS TO PREDICT FATALITIES IN HELICOPTER EMERGENCY MEDICAL SYSTEMS (HEMS) CRASHES

Background: In the past several decades, multiple studies have examined factors influencing occupant survival in aviation crashes, but only a few have addressed this question in Helicopter Emergency Medical Systems (HEMS) accidents. The four-point FIA Score is a valid tool to measure fatality risk in aviation crashes. Objective: The aim of the present study was to analyze the performance of the FIA Score when applied to German HEMS accidents, and to determine the prognostic value for fatalities and for survival. Methods: The FIA Score uses three parameters (F = fire; I = Instrument meteorological conditions; A = Away from airport) to determine the fatality risk after crashes. Data for German HEMS accidents between October 1970 and December 2009 were gathered retrospectively from the Federal Agency for Flight Accident Investigation. Accidents were graded according to the FIA Score by two emergency physician-pilots, and crash fatality rates (CFR) were calculated. Results: A total of 99 HEMS accidents were analyzed that occurred from September 1970 to December 2009. In 19.2% of these, at least one occupant was fatally injured. There were 63 accidents (63.6%) that took place with no injuries; 8 occurred with minor injuries (8.1%); and 9 resulted in major injuries (9.1%). A total of 72 data sets were complete and were used for analysis. Depending on the FIA Score (FIA0, FIA1, FIA2, and FIA3), CFRs of 0.0%, 8.1%, 53.3%, and 100.0%, respectively, were calculated. Conclusions: The FIA Score is a valuable tool in German HEMS accident analysis. It can predict fatalities and is easy to use. Thus, it may also be a valuable tool in EMS call centers to predict survival after a crash. (C) 2012 Elsevier Inc

Transcutaneous PtcCO(2) measurement in combination with arterial blood gas analysis provides superior accuracy and reliability in ICU patients

Hyper or hypoventilation may have serious clinical consequences in critically ill patients and should be generally avoided, especially in neurosurgical patients. Therefore, monitoring of carbon dioxide partial pressure by intermittent arterial blood gas analysis (PaCO2) has become standard in intensive care units (ICUs). However, several additional methods are available to determine PCO2 including end-tidal (PetCO(2)) and transcutaneous (PtcCO(2)) measurements. The aim of this study was to compare the accuracy and reliability of different methods to determine PCO2 in mechanically ventilated patients on ICU. After approval of the local ethics committee PCO2 was determined in n = 32 ICU consecutive patients requiring mechanical ventilation: (1) arterial PaCO2 blood gas analysis with Radiometer ABL 625 (ABL; gold standard), (2) arterial PaCO2 analysis with Immediate Response Mobile Analyzer (IRMA), (3) end-tidal PetCO(2) by a Propaq 106 EL monitor and (4) transcutaneous PtcCO(2) determination by a Tina TCM4. Bland-Altman method was used for statistical analysis; p < 0.05 was considered statistically significant. Statistical analysis revealed good correlation between PaCO2 by IRMA and ABL (R-2 = 0.766; p < 0.01) as well as between PtcCO(2) and ABL (R-2 = 0.619; p < 0.01), whereas correlation between PetCO(2) and ABL was weaker (R-2 = 0.405; p < 0.01). Bland-Altman analysis revealed a bias and precision of 2.0 +/- 3.7 mmHg for the IRMA, 2.2 +/- 5.7 mmHg for transcutaneous, and -5.5 +/- 5.6 mmHg for end-tidal measurement. Arterial CO2 partial pressure by IRMA (PaCO2) and PtcCO(2) provided greater accuracy compared to the reference measurement (ABL) than the end-tidal CO2 measurements in critically ill in mechanically ventilated patients patients

Helicopter Type and Accident Severity in Helicopter Emergency Medical Services Missions

HINKELBEIN J, SCHWALBE M, WETSCH WA, SPELTEN O, NEUHAUS C. Helicopter type and accident severity in Helicopter Emergency Medical Services missions. Aviat Space Environ Med 2011; 82:1148-52. Objective: Whereas accident rates and fatal accident rates for Helicopter Emergency Medical Services (HEMS) were investigated sufficiently, resulting consequences for the occupants remain largely unknown. The present study aimed to classify HEMS accidents in Germany to prognosticate accident severity with regard to the helicopter model used. Methods: German HEMS accidents (1 Sept. 1970-31 Dec. 2009) were gathered as previously reported. Accidents were categorized in relation to the most severe injury, i.e., 1) no; 2) slight; 3) severe; and 4) fatal injuries. Only helicopter models with at least five accidents were analyzed to retrieve representative data. Prognostication was estimated by the relative percentage of each injury type compared to the total number of accidents. Results: The model BO105 was most often involved in accidents (38 of 99), followed by BK117 and UH-1D. Of N = 99 accidents analyzed, N = 63 were without any injuries (63.6%), N = 8 resulted in minor injuries of the occupants (8.1%), and N = 9 in major injuries (9.1%). Additionally, N = 19 fatal accidents (19.2%) were registered. EC135 and BK117 had the highest incidence of uninjured occupants (100% vs. 88.2%) and the lowest percentage of fatal injuries (0% vs. 5.9%; all P > 0.05). Most fatal accidents occurred with the models UH-1D, Bell 212, and Bell 412. Discussion: Use of the helicopter models EC135 and BK117 resulted in a high percentage of uninjured occupants. In contrast, the fatality rate was highest for the models Bell UH-1D, Bell 222, and Bell 412. Data from the present study allow for estimating accident risk in HEMS missions and prognosticating resulting fatalities, respectively