A novel tool for stable nitrogen isotope analysis in aqueous samples.

RATIONALE: Bulk stable isotope analysis (BSIA) of dissolved matter (e.g. dissolved organic carbon, total nitrogen bound (TNb ), etc.) is of particular importance since this pool is a prime conduit in the cycling of N and C. Studying the two elemental pools is of importance, as transformation and transport processes of N and C are inextricably linked in all biologically mediated systems. No system able to analyze natural abundance stable carbon and nitrogen isotope composition in aqueous samples (without offline sample preparation) and simultaneously has been reported so far. Extension of the high-temperature combustion (HTC) system, to be capable of measuring TNb stable nitrogen isotope composition, is described in this study. METHODS: To extend the TOC analyzer to be capable of measuring TNb , modifications from the HTC high-performance liquid chromatography/isotope ratio mass spectrometry (HPLC/IRMS) interface were implemented and expanded. A reduction reactor for conversion of NOx into N2 was implemented into the new developed system. The extension addresses mainly the development of the focusing unit for nitrogen and a degassing device for online separation of TNb from molecular nitrogen (N2 ) prior to injection. RESULTS: The proof of principle of the system was demonstrated with different compound solutions. In this initial testing, the δ15 NAIR-N2 values of the tested compounds were determined with precision and trueness of typically ≤0.5‰. Good results (u ≤ 0.5‰) could be achieved down to a TNb concentration of 40 mgN/L and acceptable results (u ≤ 1.0‰) down to 5 mgN/L. In addition, the development resulted in the first system reported to be suitable for simultaneous and direct δ13 C and δ15 N BSIA of aqueous samples. CONCLUSIONS: The development resulted in the first system shown to be suitable for both δ13 C and δ15 N direct BSIA in aqueous samples. This system could open up new possibilities in SIA-based research fields. Copyright © 2016 John Wiley & Sons, Ltd.

Transplanted human cord blood-derived unrestricted somatic stem cells improve left-ventricular function and prevent left-ventricular dilation and scar formation after acute myocardial infarction.

OBJECTIVE: Functional improvement after acute myocardial ischaemia (MI) has been achieved by transplantation of different adult stem and progenitor cell types. It is controversial whether these cell types are able to form novel functional myocardium. Alternatively, graft-related or immune-related paracrine mechanisms may preserve existing myocardium, improve neovascularisation, affect tissue remodelling or induce endogenous de novo formation of functional myocardium. We have applied an alternative somatic cell type, human cord-blood-derived unrestricted somatic stem cells (USSCs) in a porcine model of acute MI. METHODS: USSCs were transplanted into the acutely ischaemic lateral wall of the left ventricle (LV). LV dimension and function were assessed by transoesophageal echocardiography (TEE) pre-MI, immediately post-MI, 48 hours and 8 weeks after USSC injection. Additionally, apoptosis, mitosis and recruitment of macrophages were examined 48 hours post-engraftment. RESULTS: Gender-specific and species-specific FISH/immunostaining failed to detect engrafted donor cells 8 weeks post-MI. Nevertheless, cell treatment effectively preserved natural myocardial architecture. Global left ventricular ejection fraction (LVEF) before MI was 60% (7%). Post-MI, LVEF decreased to 34% (8%). After 8 weeks, LVEF had further decreased to 27% (6%) in the control group and recovered to 52% (2%) in the USSC group (p<0.01). Left-ventricular end-diastolic volume (LVEDV) before MI was 28 (2) ml. 8 weeks post-MI, LVEDV had increased to 77 (4) ml in the control group. No LV dilation was detected in the USSC group (LVEDV: 26 (2) ml, p<0.01). Neither apoptosis nor recruitment of macrophages and mitosis were different in either groups. CONCLUSIONS: Transplantation of USSCs significantly improved LV function and prevented scar formation as well as LV dilation. Since differentiation, apoptosis and macrophage mobilisation at infarct site were excluded as underlying mechanisms, paracrine effects are most likely to account for the observed effects of USSC treatment.

Sheep and goats: separating cervix and corpus uteri from imprecisely coded uterine cancer deaths, for studies of geographical and temporal variations in mortality.

Analysing time trends in mortality from cancers of the cervix and corpus uteri using routine data sources (such as the World Health Organisation mortality database) involves two major problems: deaths certified as "uterus, unspecified site", and the presence of a combined category comprising unspecified and corpus uteri cancer deaths. To avoid misleading interpretations, the unspecified and the misclassified data must be incorporated into the analysis to produce rates that allow meaningful comparisons between populations and over time. Reallocation methods based on age- and time-specific distributions of cervix and corpus uteri cancer are applied to the unspecified deaths, while for those in the combined category, the age- and time-specific distributions of unspecified and corpus uteri cancer are considered. Adjustments of the general strategies for reallocation were developed to take into account the different quality of the data. Results from eight European countries with different degrees of coding precision are presented. The reallocation methods bring the cervix and corpus uteri mortality trends more in line with the trends for countries with more precise data while keeping the country-specific characteristics. In addition, the methods ensured the availability of time trends for corpus uteri cancer in women age 50 years and older, which were completely missing without reallocation. We propose generally applicable reallocation methods that allow valid time trend analysis of cervix and corpus uteri cancer mortality using datasets of varying precision. Our results show that any sensible analysis of time trends must involve procedures for correcting for unspecified and misclassified uterine cancer deaths. The modified data are available at .

[Reconstruction of the extensor pollicis longus tendon by transposition of the extensor indicis tendon]. / Rekonstruktion der Extensor pollicis longus-Sehne mittels Extensor indicis-Transposition.

Rupture of the extensor pollicis longus-tendon (EPL) is a frequent complication after distal radius fractures. Other traumatic and non-traumatic reasons for this tendon lesion are known, including a theory about a disorder in the blood supply to the tendon itself. We examined 40 patients after reconstruction of the EPL-tendon in a mean follow-up time of 30 months. All patients were clinically examined and a DASH questionnaire was answered by all patients. The method to reconstruct the EPL-tendon was the transposition of the extensor indicis-tendon. After the operations the thumb was put in a splint for four weeks in a "hitch-hiker's-position". 31 ruptures of the tendon (77.5 %) were a result of trauma. In 20 of them (50 %) a distal radius fracture had occurred. Clinical examination included measurements of the movement of the thumb- and index-finger joints, the grip strength and the maximal span of the hand. Significant differences were not found. The isolated extension of the index finger was possible in all patients. But it was reduced in ten cases which represent 25 %. Our results were evaluated by the Geldmacher score to evaluate the reconstruction of the EPL-tendon. 20 % excellent, 65 % good, 12.5 % fair and 2.5 % poor results were reached. The Geldmacher score was used critically. We suggest its modification for the evaluation of thumb abduction. The DASH score reached a functional value of ten points which represents a very good result. In conclusion the extensor indicis-transposition is a safe method to reconstruct the EPL-tendon. Its substantial advantage is taking a healthy muscle as the motor, thereby avoiding the risk of using a degenerated muscle in late tendon reconstruction. A powerful extension of the index finger will be maintained by physical education. Generally, the loss of the extension of the index finger is negligible. It does not disturb the patients. But it has to be discussed with the patient before the operation.

Risk factors influencing the outcome after surgical treatment of complicated deep sternal wound complications.

BACKGROUND: Median sternotomy is the most frequently used incision for cardiac procedures but carries a substantial risk for deep sternal wound infections and/or sternal dehiscence. In contrast to previous studies that examined risk factors for sternal infections this study evaluates factors that lead to poor outcome after surgical revision of the non healing sternum. METHODS: Between 1985 and 1999, 193 adults (mean age 64 +/- 9 years, m/f = 3/1) necessitated sternal revisions (incidence 1.93%). Pre-, intra- and post-operative risk factors were evaluated for their influence on the outcome after sternal revision. RESULTS: 65 of the 193 patients had a complicated course: ten (5.2%) died due to sepsis/multi organ failure (n = 6) or cardiac causes (n = 4). 32 patients (16.6%) needed several revisions, 17 (9%) were discharged with sternal instability, 5 (3%) with chronic fistula and one with persistent osteomyelitis. Univariate and multivariate analysis identified cardiopulmonary resuscitation (odds ratio (OR) = 11.188, p = 0.010), corticoid treatment (OR = 7.043, p = 0.0055), diabetes (OR = 4.130, p = 0.0128), smoking history (OR = 2.996, p = 0.0041), renal insufficiency (hazard ratio (HR) = 1.884), old age (OR = 1.108, p = 0.0266), high body mass (HR = 1.06), ECC time (p = 0.023), cross clamp time (p = 0.028), systemic hypothermia (p = 0.016), non-use of IMA (p = 0.042) or prolonged ventilation as risk factors for mortality or poor outcome. No correlation between sternal closure technique, mediastinal irrigation or antibiotic therapy and outcome after mediastinal revision could be found. CONCLUSIONS: To avoid disappointing results after sternal revision one should aim to preoperatively identify high-risk patients and aggressively address risk factors. This rather than modifications of the surgical and medical approach might improve the outcome of patients with mediastinal complications.

Disparity between changes in mRNA abundance and enzyme activity in Corynebacterium glutamicum: implications for DNA microarray analysis.

The relationship between changes in mRNA abundance and enzyme activity was determined for three genes over a span of nearly 3 h during amino acid production in Corynebacterium glutamicum. Gene expression changes during C. glutamicum fermentations were examined by complementary DNA (cDNA) microarrays and by a second method for quantitating RNA levels, competitive reverse transcriptase-PCR (RT-PCR). The results obtained independently by both methods were compared and found to be in agreement, thus validating the quantitative potential of DNA microarrays for gene expression profiling. Evidence of a disparity between mRNA abundance and enzyme activity is presented and supports our belief that it is difficult to generally predict protein activity from quantitative transcriptome data. Homoserine dehydrogenase, threonine dehydratase, and homoserine kinase are enzymes involved in the biosynthesis of l-isoleucine and other aspartate-derived amino acids in C. glutamicum. Our data suggest that different underlying regulatory mechanisms may be connected with the expression of the genes encoding each of these three enzymes. Indeed, whereas in one case the increases in enzyme activity exceeded those in the corresponding mRNA abundance, in another case large increases in the levels of gene expression were not congruent with changes in enzyme activity.

[Coronary revascularization: off-pump versus on-pump--a comparison of behavior of biochemical cardiac ischemia markers]. / Koronarrevaskularisation: Off-pump versus On-pump--Verhalten der biochemischen kardialen Ischämiemarker im Vergleich.

BACKGROUND: Recently, coronary artery bypass grafting (CABG) on the beating heart with avoidance of extracorporeal circulation (off-pump CABG technique) has been gaining increasing importance in modern cardiac surgery. The object of this prospective study was to compare postoperative kinetic and patterns of cardiac troponin I (cTnI), T (cTnT), and creatine kinase MB (CKMB) activities after off-pump CABG versus conventional on-pump CABG. METHODS: We studied 106 patients who underwent first-time elective on-pump (group I, n = 69, 56 male, 13 female, mean age: 64.3 +/- 9.9 years, mean ejection fraction: 56 +/- 15%) or off-pump (group II, n = 37, 24 male, 13 female, mean age: 68.4 +/- 9.1 years, mean ejection fraction: 57 +/- 13%) CABG surgery via median sternotomy. CTn I and cTnT levels, total creatine kinase (CK) and CK-MB activities in the serum were measured before operation, up on arrival at the ICU and 6, 12, 24, 48 and 120 hours later. Serial 12-lead ECGs were recorded preoperatively and on days 1, 2 and 5. RESULTS: Serum concentrations of cardiac troponins in all patients were preoperatively either not detectable or in the normal range and significantly increased after surgery. In group I, one patient developed a Q wave myocardial infarction, one patient a non-Q wave infarction and two patients a new left bundle branch block on the ECG. One patient of group II developed a new Q-wave myocardial infarction and another patient permanent atrial fibrillation associated with a continuous arrhythmia. All patients with a myocardial infarction in the ECG showed significant elevation of concentrations or activities of these biochemical markers. The median postoperative peak values for cTnI were measured at 24 h in both groups (2.7 micrograms/l, 95%-CI: [2.2, 3.2] in group I and 1.1 micrograms/l, 95%-CI: [0.5, 1.3] in group II). CTnT postoperatively presented an earlier median peak of 0.128 microgram/l at 12 h in group II (95%-CI: [0.041, 0.146]) than in group I at 48 h (0.298 microgram/l, 95%-CI: [0.254, 0.335]). CONCLUSIONS: All patients undergoing CABG surgery with or without extracorporeal circulation postoperatively showed an increase of cardiac troponin levels. After uncomplicated coronary revascularization, patients with the off-pump CABG technique continuously presented lower serum cardiac troponin concentrations than those with the on-pump CABG technique. CTnI showed the same patterns of release in both groups with different median postoperative peak values at 24 h. The patterns off cTnT release following CABC surgery with or without extracorporal circulation were different: CTnT reaches its postoperative peak value in patients with the off-pump CABG technique earlier than those with the on-pump CABG technique (12 h postoperatively versus 48 h).

Correlation of platelet concentration in platelet-rich plasma to the extraction method, age, sex, and platelet count of the donor.

An important reason to improve methods of isolating platelet-rich plasma (PRP) is the potential use of autologous platelet growth factors. In addition to discontinuous plasma separation, a second method for extraction of PRP has now become available, which can be performed directly by the surgeon. In this study, the suitability of the 2 methods of producing PRP was compared. Whole blood was drawn from 158 healthy donors (112 men, 46 women) aged 20 to 62 years (mean 34, SD 10). The PRP was separated by the discontinuous plasma separation method (by the blood bank) or by the so-called "buffy coat" method (the "self-concentration" method, analogous to the PRP Kit, Curasan, Kleinostheim, Germany). Platelet counts differed significantly according to donor blood (median men 237,500/microL, women 272,000/microL), blood bank PRP preparation (median men 1,302,000/microL, women 1,548,500/microL), and self-concentrated PRP (median men 944,000/microL, women 1,026,000/microL). The platelet concentration of the blood bank PRP correlated with the platelet count in the donor whole blood (Spearman's correlation coefficient r(s) = 0.73). However, there was no significant correlation between the platelet count of self-concentrated PRP and donor whole blood (r(s) = 0.22). Significant but irrelevant influences of sex on platelet concentration were found, but no influence of age was detected.

Patterns and diagnostic value of cardiac troponin I vs. troponin T and CKMB after OPCAB surgery.

BACKGROUND: Cardiac troponin I (cTnI) has been shown to be a specific marker for myocardial injury in cardiac surgery. The object of this prospective study was to determine the patterns and kinetic and diagnostic value of cTnI, cardiac troponin T (cTnT), and creatine kinase MB (CKMB) activity after minimally invasive coronary revascularization using an octopus device on the beating heart (OPCAB). METHODS: 48 patients (33 male/15 female, mean age 68.3 +/- 8.7 years) underwent their first elective OPCAB surgery with median sternotomy without mortality. The mean number of grafts was 2.0 +/- 0.8 per patient. Preoperative mean ejection fraction was 56.6 % +/- 14.9%. CTnI and T levels, total creatine kinase (CK) and CK-MB activity in the serum were measured before operation, at arrival at the ICU, and 6, 12, 24, 48 and 120 hours afterward. Serial 12-lead ECGs were recorded preoperatively and at days 1, 2 and 5. The relationship between perioperative data and postoperative cTnI and cTnT levels and CKMB were statistically identified for all variables. RESULTS: The best cutoff value for cTnI was 8.35 micrograms/l. The patients were grouped by the ECG findings and maximal slopes of cTnI postoperatively (group I: unchanged ECG and cTnI < 8.35 micrograms/l, n = 38; group II: unchanged ECG and cTnI > 8.35 micrograms/l n = 6; group III: Q-wave in ECG and cTnI > 8.35 micrograms/l, n = 4). Baseline serum concentrations of cTnI were in the normal range, and significantly increased after surgery with a peak 24h after the operation. Maximal slopes of cTnI ranged in group II between 9.1 and 18.0 micrograms/l, and in group III between 35.9 and 88.8 micrograms/l. There was strong concordance between maximum cTnI, cTnT (p < 0.0001) and CK-MB levels (p = 0.003). First cTnI levels immediately post-op correlated with the maximum cTnI levels during the postoperative course (p = 0.009). CONCLUSIONS: CTnI after minimal invasive surgery shows a characteristic pattern with a maximum at 24h after the operation. The measurement of postoperative biochemical marker concentrations, specially cTnI, reflects myocardial injury incurred during the procedure. It is an accurate method for confirming or excluding a perioperative myocardial injury diagnosis after OPCAB surgery.

[IMZ-TwinPlus bone implant system--4 years clinical experience]. / Das IMZ-TwinPlus-Schraubenimplantat-System--4 Jahre klinische Erfahrung.

AIM OF THE STUDY: For more than twenty years the IMZ-system has been in clinical use, since 1995 modified as IMZ-TwinPlus. The aim of this prospective clinical trial was to analyse the success of the latter implant system, which is an automatically threading cylindrical titanium screw with a deep structured surface (Fa. FRIATEC AG, Mannheim, BRD). Investigated parameters were the peri-implant situation of the soft tissues, the bone loss and the rate of implant failure after a maximum period of 4.5 years of clinical use. MATERIAL AND METHODS: From 1995-2000 sixty-eight patients were provided with a total of 278 IMZ TwinPlus screw implants for various indications (mainly alveolar ridge atrophy). 191 implants were inserted without any, 35 implants with a loco-regional and another 52 implants after comprehensive reconstructive osteoplastic surgery. 38 patients with 155 implants were re-examined using a standardised protocol to evaluate peri-implant hard- and soft tissue situation as well as the patient's subjective assessment of the treatment. RESULTS: With a maximum of 54 months the mean observation period was 30 months. The implant failure rate has risen to 6% so far (n = 18 in 12 patients). Two patients bearing 7 implants have passed away. One patient with 4 implants was lost to follow up. 249 implants were still under function at examination, thus the in situ rate was 91%. The Kaplan-Meier survival rate after 54 months proved 91%. DISCUSSION: To our knowledge there are at present no other data published on the survival rate of the IMZ TwinPlus implant system. The results of this study evaluate a survival rate similar to the classical IMZ cylinder implant and other implant systems for the analysed observation period. CONCLUSIONS: After a maximum observation period of 4.5 years the IMZ TwinPlus implant system showed results in the range of other well-established implant systems. Designed to resist rotation of the superstructure the IMZ TwinPlus screw implant widens the range of indications in comparison to the classical IMZ cylinder implant system.

Development and validation of corynebacterium DNA microarrays.

We have developed DNA microarray techniques for studying Corynebacterium glutamicum. A set of 52 C. glutamicum genes encoding enzymes from primary metabolism was amplified by PCR and printed in triplicate onto glass slides. Total RNA was extracted from cells harvested during the exponential-growth and lysine production phases of a C. glutamicum fermentation. Fluorescently labeled cDNAs were prepared by reverse transcription using random hexamer primers and hybridized to the microarrays. To establish a set of benchmark metrics for this technique, we compared the variability between replicate spots on the same slide, between slides hybridized with cDNAs from the same labeling reaction, and between slides hybridized with cDNAs prepared in separate labeling reactions. We found that the results were both robust and statistically reproducible. Spot-to-spot variability was 3.8% between replicate spots on a given slide, 5.0% between spots on separate slides (though hybridized with identical, labeled cDNA), and 8.1% between spots from separate slides hybridized with samples from separate reverse transcription reactions yielding an average spot to spot variability of 7.1% across all conditions. Furthermore, when we examined the changes in gene expression that occurred between the two phases of the fermentation, we found that results for the majority of the genes agreed with observations made using other methods. These procedures will be a valuable addition to the metabolic engineering toolbox for the improvement of C. glutamicum amino acid-producing strains.

Postoperative patterns and kinetics of cTnI, cTnT, CK-MB-activity and CK-activity after elective aortic valve replacement.

OBJECTIVES: The aim of this prospective study was to evaluate postoperative kinetics of four different biochemical ischaemic markers after elective aortic valve replacement (AVR). Additionally, pre-, peri- and postoperative data were analysed in order to identify factors with possible impact on the postoperative release of the selected enzymes. DESIGN: Forty patients (14 males, 26 females, aged 70 +/- 11 years; EF = 54 +/- 18% [mean +/- SD]) undergoing elective AVR were prospectively included in this study. For all patients, serum concentrations of cTnI, cTnT, and serum activities of CK-MB and CK were measured preoperatively as well as 0, 6, 12, 24, 48 and 120 hours after removal of the aortic cross-clamp. Clinical data were assessed in all patients and correlated with postoperative enzyme patterns. RESULTS: There were no major complications. Preoperatively, all patients showed enzyme values in the normal range whereas the four ischaemic markers reached higher values postoperatively. cTnI reached its maximum values 24 hours (XMed = 2.35 micrograms/L, 95%-CI [2.0, 3.3]) and cTnT 48 hours after the operation (XMed = 0.239 microgram/L, 95%-CI [0.174, 0.283]). Typical biphasic release kinetics could be demonstrated for cTnT. There was a high linear correlation between cTnI and cTnT at all sampling times. In contrast, a high linear correlation between cTnI, cTnT, and CK-MB-activity was only found 48 hours after aortic unclamping. cTnI nearly was in normal range 120 h postoperatively (XMed = 0.5 microgram/L, 95%-CI [0.2, 0.6]), whereas cTnT still remained pathologically elevated (XMed = 0.223 microgram/L, 95%-CI [0.137, 0.299]). No linear correlation was found between maximum values of the ischaemic markers postoperatively and age, gender, body surface area, ejection fraction, LV-hypertrophy, operating time, ECC time, time of cardiac arrest, lowest body temperature, perfusion pressure, cardioplegia volume, reperfusion time, postoperative septiformic circulatory instability, or ventilation time. CONCLUSIONS: All four ischaemic markers showed individual peak characteristics and kinetics after uncomplicated AVR. In contrast to previous findings, aortic cross-clamping time had no detectable impact on postoperative peak patterns of any ischaemic marker.

Changes in cellular proliferation rate of lymphocytes after long-distance flights as a possible risk for patients with HIV-infection.

OBJECTIVES: Several studies showed that long-distance flights can influence cellular immunity. This might be due to a cortisol- and catecholamin- induced change in immunity with an impairment of T-lymphocyte dependent cellular immunity and an enhancement of B-lymphocyte dependent humoral immunity. Similar results can be found in patients with HIV. It is also known that progress of this disease and affection of T-helper-cells by the virus are induced by stimulation of the immune system, a phenomenon that also occurs during long distance flights. Therefore, a possible interaction between long-distance flights and the progression of HIV-infection should be discussed. METHODS: Cell cultures of 22 subjects after long-distance flights with and without rapid time zone shifts and of 16 patients with HIV (stage 2 3) were investigated. Mononuclear blood cells were stimulated with different lectins in culture and proliferation was measured by incorporation of bromodesoxyuridine. Moreover, all cultures were titrated with chromate concentrations between 0 to 700 ng/ml to measure the tolerance of the cells against chromate (VI) in vitro as a marker of the functional efficiency of the cellular part of the immune system. Maximal proliferation rate and tolerance against chromate were compared in both groups. RESULTS: After long distance flights tolerance against chromate decreased significantly during the first 24 h after flight. After 48 h levels were similar to those 1 week after flights. The decrease was similar to the results found in the stage 2 group of HIV-patients, but by far less to the decrease in stage 3 patients. Maximal proliferation rate dropped significantly during the second day after arrival compared to 1-week control values. CONCLUSION: Changes in the cellular immune system in healthy subjects after long-distance flights have been similar to the results of patients with stage 2 of HIV-infection. Mechanisms of changes in both groups are comparable in influencing T-cell-induced immunity. This could point to an additive effect on cellular immunity of HIV-patients by long distance-flights. Rosen neopterin concentrations and increases of apoptotic T-cells in both groups support this assumption. Therefore, further studies are urgently needed to investigate the interactions between HIV-infection and long-distance flights.

VEGF(165) mediates glomerular endothelial repair.

VEGF(165), the most abundant isoform in man, is an angiogenic cytokine that also regulates vascular permeability. Its function in the renal glomerulus, where it is expressed in visceral epithelial and mesangial cells, is unknown. To assess the role of VEGF(165) in glomerular disease, we administered a novel antagonist - a high-affinity, nuclease-resistant RNA aptamer coupled to 40-kDa polyethylene glycol (PEG) - to normal rats and to rats with mesangioproliferative nephritis, passive Heymann nephritis (PHN), or puromycin aminonucleoside nephrosis (PAN). In normal rats, antagonism of VEGF(165) for 21 days failed to induce glomerular pathology or proteinuria. In rats with mesangioproliferative nephritis, the VEGF(165) aptamer (but not a sequence-scrambled control RNA or PEG alone) led to a reduction of glomerular endothelial regeneration and an increase in endothelial cell death, provoking an 8-fold increase in the frequency of glomerular microaneurysms by day 6. In contrast, early leukocyte influx and the proliferation, activation, and matrix accumulation of mesangial cells were not affected in these rats. In rats with PHN or PAN, administration of the VEGF(165) aptamer did not influence the course of proteinuria using various dosages and administration routes. These data identify VEGF(165) as a factor of central importance for endothelial cell survival and repair in glomerular disease, and point to a potentially novel way to influence the course of glomerular diseases characterized by endothelial cell damage, such as various glomerulonephritides, thrombotic microangiopathies, or renal transplant rejection.

Protein binding regions of the mRNAs for the 55 kDa tumor necrosis factor receptor and the glucose transporter 1: sequence homology and competition for cellular proteins.

Gene expression is influenced by mechanisms regulating mRNA degradation. Knowledge on regulatory RNA elements involved and on proteins interacting with them is still limited. A 33 nucleotide (nt) region of the 55 kDa tumor necrosis factor receptor (TNFR-55) mRNA, previously reported by us to engage in such interaction with proteins from U-937 cells, exhibits homology to a 38 nt regulatory region of the glucose transporter GLUT-1 mRNA. Labeled RNA fragments comprising these two regions bind similar sets of proteins. Upon phorbol ester-induced differentiation into macrophage-like cells, protein binding to both fragments is changed similarly. Furthermore, both compete with each other for protein binding. This suggests that GLUT-1 and TNFR-55 RNA share a novel protein binding RNA motif involved in regulation of their half life.

Detection of 100% of the CFTR mutations in 63 CF families from Tyrol.

We identified 100% of the CFTR gene mutations, including three novel mutations, in 126 unrelated cystic fibrosis chromosomes from Tyrol, Austria. The frequency of the major mutation deltaF508 (74.6%) was not significantly different in Tyrolian CF-patients than in patients from Bavaria (71.0%) and Middle- and Northern Germany (71.9%), but was significantly higher than in patients from Styria (58.1%) or Northern Italy (47.6%). Interestingly, the distribution of the next most frequent mutations, R1162X (8.7%) 2183AA-->G, 2789+5G-->A and G542X (2.4% each), was more similar to the distribution of these mutations among CF-patients from Northern Italy than to those from Styria, Bavaria or Middle- and Northern Germany. Nine further mutations occurred once or twice. One of these, the missense mutation M1101K, is rare worldwide but very frequent in the Hutterite brethren, a small founder population which came from Southern Austria to Northern America. Three other different mutations (deltaL453, 1874insT and 4108delT) were present in single Tyrolian families and have not been described before. The identification of 100% of CFTR gene mutations in a particular CF population demonstrates the power of genetic analysis for the diagnosis and counselling of CF families in this restricted geographical area of Austria. Our study provides evidence for a closer genetic relation between CF patients from Tyrol and those from Bavaria or Middle- and Northern Germany as well as Northern Italy, than between CF patients from the two Austrian states Tyrol and Styria.

[Intramuscular application of midazolam. Its effect upon CNS and respiration (author's transl)]. / Intramuskuläre Applikation von Midazolam. Wirkung auf ZNS und Atmung.

8-Chloro-6-(2-fluorophenyl)-1-methyl-4H-imidazo[1,5-a] [1,4]benzodiazepine (midazolam, Ro 21-3981, Dormicum) is a benzodiazepine with a pronounced sleep-inducing effect. Deep-sleep stages and respiratory depressions were observed after 5 mg i.v. Whether and in what dosage midazolam can be safely, i.e., without continuous monitoring administered i.m. must be shown by the analysis of larger study populations. After i.m. injection, there is rapid onset of action leading to sleep stages which are comparable only with those hitherto observed after i.v. injection.

[Maternal serum levels of cystine-aminopeptidase (C.A.P.) in normal and pathological pregnancies (author's transl)]. / Etude des taux sériques de cystine-aminopeptidase (C.A.P.) ou ocytocinase dans la grossesse normale et pathologique.

The variations of the maternal serum levels of cystine-aminopeptidase or oxytocinase (C.A.P.) have been studied in 399 measurements from 326 pregnant patients, 226 normal pregnancies were used as control and allowed us to set up a range of normal values for the age of pregnancy; the mean levels increase exponentially. It has not been possible from this study to establish conclusively a direct relationship between the C.A.P. levels, or variations of these levels and the onset of a fetal and/or placental pathology. In twin pregnancies only, the levels were constantly elevated.