Calafiore cardioplegia offers better myocardial protection in adult and senescent rat hearts than Del Nido cardioplegia.

OBJECTIVES: We compared the cardioprotective capacity of Del Nido cardioplegia and warm Calafiore blood cardioplegia in an experimental setting during 90 min of ischaemia. METHODS: 20 adult and 20 senescent rat hearts were isolated and mounted on a blood-perfused, pressure-controlled Langendorff apparatus. After a stabilization period, cardiac arrest (90 min) was induced by the administration of either Calafiore (Cala) or Del Nido solution (DNS). While Cala was given warm and intermittently, DNS was given as a cold single shot. During 90 min of reperfusion, cardiac function and metabolism were evaluated and biomarker levels were measured. After the end of the experiment, hearts were prepared for electronmicroscopic investigation. RESULTS: Hearts exposed to Cala recovered faster during reperfusion compared with hearts administered DNS (Cala vs DNS at 30 min reperfusion: left ventricular developed pressure 72, SD: 22% of baseline (BL) versus 40, SD: 32% of BL, p < .001, and positive derived left ventricular pressure over time was better in both adult and senescent Cala groups (96, SD: 31% of BL) than in the DNS groups (39, SD: 27% of BL, p < .001). Ischaemic contractures were seen in the DNS groups starting after 30 min of ischaemia, whereas no rise in diastolic pressure was observed for the Cala groups. Accordingly, lactate production was higher after DNS (1.23 mg/dl (SD 0.87) than after Cala (0.33 mg/dl (SD 0.68), p = .015) at the beginning of reperfusion. Troponin I levels at the end of reperfusion were higher after DNS treatment in adult hearts (DNS: 287.9 SD: 147.7 ng/mL vs Cala 91.2: SD: 94.7 ng/mL, p = .02) and in senescent hearts (DNS: 376.5 (SD: 162.8) ng/ml versus Cala 104.7 (SD: 150.2) ng/ml, p = .025). Electron microscopy showed that the cellular oedema index was higher in adult DNS hearts (1.2 ± 0.2) than in adult Cala hearts (0.8 ± 0.1, p = .012), whereas the VS ratio was similar (0.18 ± 0.01 vs 0.17 ± 0.03). CONCLUSION: Calafiore cardioplegia offers better myocardial protection from ischaemia/reperfusion-related damage in isolated perfused adult and senescent rat hearts than Del Nido cardioplegia.

[Importance of psychosomatic medicine for people with rare diseases]. / Bedeutung der psychosomatischen Medizin für Menschen mit seltenen Erkrankungen.

People with rare diseases have a very high rate of mental and social stress. This results in specific tasks and problems in the psychosomatic care of patients. On the one hand, the physical and/or psychological symptoms of an undetected rare organic disease can be misdiagnosed as a psychosomatic disease, and the affected persons possibly receive psychotherapy that is not causally effective. On the other hand, mental diseases that require treatment can arise as a result of the effects of a rare disease. These should be diagnosed as such and treated with psychotherapy. If, in individual cases, both symptoms of a rare disease and symptoms of a psychosomatic disorder in the sense of comorbidity are present, neither one nor the other diagnosis should lead to a hasty termination of diagnostic efforts. Otherwise, misalignments can easily occur and the further diagnostic and therapeutic process can be permanently disturbed. Interdisciplinary team care interventions should therefore be developed further.

Controlled, cross-sectional MRI evaluation of joint status in severe haemophilia A patients treated with prophylaxis vs. on demand.

In patients with haemophilia A, factor VIII (FVIII) prophylaxis reduces bleeding frequency and joint damage compared with on-demand therapy. To assess the effect of prophylaxis initiation age, magnetic resonance imaging (MRI) was used to evaluate bone and cartilage damage in patients with severe haemophilia A. In this cross-sectional, multinational investigation, patients aged 12-35 years were assigned to 1 of 5 groups: primary prophylaxis started at age <2 years (group 1); secondary prophylaxis started at age 2 to <6 years (group 2), 6 to <12 years (group 3), or 12-18 years (group 4); or on-demand treatment (group 5). Joint status at ankles and knees was assessed using Compatible Additive MRI scoring (maximum and mean ankle; maximum and mean of all 4 joints) and Gilbert scores in the per-protocol population (n = 118). All prophylaxis groups had better MRI joint scores than the on-demand group. MRI scores generally increased with current patient age and later start of prophylaxis. Ankles were the most affected joints. In group 1 patients currently aged 27-35 years, the median of maximum ankle scores was 0.0; corresponding values in groups 4 and 5 were 17.0 and 18.0, respectively [medians of mean index joint scores: 0.0 (group 1), 8.1 (group 2) and 13.8 (group 4)]. Gilbert scores revealed outcomes less pronounced than MRI scores. MRI scores identified pathologic joint status with high sensitivity. Prophylaxis groups had lower annualized joint bleeds and MRI scores vs. the on-demand group. Primary prophylaxis demonstrated protective effects against joint deterioration compared with secondary prophylaxis.

[Anesthesia in a patient after endoscopic lung volume reduction. First anesthesiological experiences with implanted endobronchial valves]. / Narkose bei einem Patienten mit Zustand nach endoskopischer Lungenvolumenreduktion. Erste anästhesiologische Erfahrungen mit implantierten "Lungenventilen"

Chronic obstructive pulmonary disease (COPD) is a disease with a high incidence and extensive comorbidities that make COPD a key challenge for anesthesiologists. A new treatment strategy, such as endoscopic lung volume reduction (ELVR) with implantation of endobronchial valves is a rapidly developing area which is still unknown to many anesthesiologists. This article therefore describes first experiences in a patient with five endobronchial valves in the right upper lobe who needed urgent surgery due to lumbar disc herniation with neurological impairment. After preoperative evaluation of the patient's condition, the use of bronchodilating volatile anesthetics and adjusting the ventilatory settings with long expiration times and low peak pressure in a pressure controlled mode seems favorable in these patients. Intraoperatively, the patient should be assessed with modern physiological monitoring tools to titrate the administration of anesthetic agents, opioids and myorelaxant drugs. In conclusion the care of patients with implanted endobronchial valves after ELVR does not differ from COPD patients without ELVR. A close cooperation between surgeons, anesthesiologists and internists is mandatory in the care of these patients.

IABP before cardiac surgery: clinical benefit compared to intraoperative implantation.

BACKGROUND: The aim of this study was to determine whether preoperative compared to intraoperative intra-aortic balloon counterpulsation (IABP) is advantageous regarding 30-day and 2-year survival in high-risk patients (acute myocardial infarction, severely impaired left ventricular ejection fraction (LVEF), low output syndrome) undergoing coronary surgery. METHODS: In the years 2004 to 2008, 156 consecutive patients undergoing coronary surgery with IABP support (119 preoperative, 37 intraoperative IABP) were observed. Applying Fisher's exact test, as well as Wilcoxon and median tests, possible group differences were evaluated. After univariate analysis, models of logistic regression and Cox-regression were built. RESULTS: Preoperative hemodynamic state and risk profile of the two patient groups were comparable: patients with preoperative IABP showed a similar level of urgency (21.9% vs. 18.9% emergencies), cardiogenic shock (8.4% vs. 10.8%), inotropes (8.4% vs. 8.1%), impaired LVEF (30.3% vs. 29.7%) and ventilation (5.9% vs. 5.4%) compared to patients with intraoperative IABP. Nevertheless, patients with intraoperative IABP demonstrated a significantly higher 30-day mortality rate (37.8% vs. 5.9%) and 2-year mortality rate (54.0% vs. 18.1%) compared to patients with preoperative IABP. Logistic regression revealed that patients with intraoperative IABP have a 16-times higher 30-day mortality rate after coronary surgery (OR: 16.386, 95% CI: 4.858-55.266) than patients with preoperative IABP. Two-year mortality (OR: 9.317, 95% CI: 3.430 to 25.311) and survival time were significantly better in patients with preoperative IABP therapy. CONCLUSION: Considering the significant benefit for patients with preoperative compared to intraoperative IABP and the absence of vascular problems after IABP insertion, the results of this study indicate a more liberal indication for IABP in high-risk patients before coronary bypass surgery.

Long-term results of mechanical and biological heart valves in dialysis and non-dialysis patients.

INTRODUCTION: We wanted to answer the question whether biological heart valves are inferior compared to mechanical heart valves in end-stage renal disease (ESRD) patients. METHODS: Between 01/1996 und 12/2006, 44 of 3293 patients undergoing aortic valve replacement (AVR) in a single institution suffered from dialysis-dependent ESRD and underwent a follow-up investigation after 1.9 years (median). Twelve (28.9 %) of these patients received a biological, 32 (71.1 %) of these patients a mechanical aortic valve prosthesis. To evaluate a possible influence of the valve type (biological/mechanical) on survival, uni- and multivariate logistic regression was used. RESULTS: ESRD patients after AVR had a relatively poor short-term (30-day mortality: 22.7 %) and long-term survival (median survival time: 24.7 months; 95 % CI: 0.2-47.7 months), irrespective of the type of heart valve prosthesis (hazard ratio for mortality depending on heart valve type in dialysis patients: 1.31, P = 0.400). Dialysis-dependent patients were not reoperated due to valve-related reasons. CONCLUSIONS: The long-term survival of dialysis-dependent patients after AVR is low (5-year survival: 29.5 %) irrespective of the type of heart valve prosthesis. Therefore, the use of biological AVR is not contraindicated in this group of patients.

Sodium/hydrogen exchange inhibition with cariporide reduces leukocyte adhesion via P-selectin suppression during inflammation.

BACKGROUND AND PURPOSE: The Na(+)/H(+) exchange (NHE) inhibitor cariporide is known to ameliorate ischaemia/reperfusion (I/R) injury by reduction of cytosolic Ca(2+) overload. Leukocyte activation and infiltration also mediates I/R injury but whether cariporide reduces I/R injury by affecting leukocyte activation is unknown. We studied the effect of cariporide on thrombin and I/R induced leukocyte activation and infiltration models and examined P-selectin expression as a potential mechanism for any identified effects. EXPERIMENTAL APPROACH: An in vivo rat mesenteric microcirculation microscopy model was used with stimulation by thrombin (0.5 micro ml(-1)) superfusion or ischaemia (by haemorrhagic shock for 60 min) and reperfusion (90 min). KEY RESULTS: Treatment with cariporide (10 mg kg(-1) i.v.) significantly reduced leukocyte rolling, adhesion and extravasation after thrombin exposure. Similarly, cariporide reduced leukocyte rolling (54+/-6.2 to 2.4+/-1.0 cells min(-1), P<0.01), adherence (6.3+/-1.9 to 1.2+/-0.4 cells 100 microm(-1), P<0.01) and extravasation (9.1+/-2.1 to 2.4+/-1.1 cells per 20 x 100 microm perivascular space, P<0.05), following haemorrhagic shock induced systemic ischaemia and reperfusion. The cell adhesion molecule P-selectin showed a profound decrease in endothelial expression following cariporide administration in both thrombin and I/R stimulated groups (35.4+/-3.2 vs 14.2+/-4.1% P-selectin positive cells per tissue section, P<0.01). CONCLUSIONS AND IMPLICATIONS: The NHE inhibitor cariporide is known to limit reperfusion injury by controlling Ca(2+) overload but these data are novel evidence for a vasculoprotective effect of NHE inhibition at all levels of leukocyte activation, an effect which is likely to be mediated at least in part by a reduction of P-selectin expression.

Increased frequency of the CTLA-4 49 A/G polymorphism in patients with acquired haemophilia A compared to healthy controls.

Acquired haemophilia (AH) is an autoimmune disorder characterized by autoantibodies against endogenous factor VIII (FVIII). Half of the patients present with an underlying disease known to cause the FVIII autoantibodies whereas in the other half the disease is of idiopathic nature. Recently, it has been shown that variants of the polymorphic cytotoxic T lymphocyte antigen-4 (CTLA-4) gene are associated with autoimmune diseases and also represent a risk factor for inhibitor formation in inherited haemophilia A. In the present study, we investigated whether CTLA-4 variants also play a role in the pathogenesis of AH. Therefore, we analyzed three single nucleotide polymorphisms (SNPs) of the CTLA-4 gene (-318 C/T, +49 A/G and CT60 A/G) in 57 AH patients and 98 controls. The CTLA-4 + 49 G allele occurred with a significantly higher frequency in patients with AH compared with controls [odds ratio (OR) = 2.17, 95% confidence interval (CI): 1.36-3.48, P = 0.001]. This effect was mainly caused by a higher frequency of the 49 G allele in female patients (OR = 5.1, 95% CI: 1.76-15.02, P = 0.002), whereas in males the frequencies were not significantly different (OR = 1.4, P = 0.5). A higher frequency of the G allele was also observed in the subcohort with AH and underlying autoimmune disease (OR = 3.1, P = 0.04). Our observations of a higher frequency of the CTLA-4 + 49 A/G SNP in AH patients are in concordance with findings in other autoimmune disorders. In conclusion, on the background of the CTLA-4 gene polymorphism, further genetic and/or environmental factors might contribute to and finally trigger the clinical manifestation of AH.

[Backpain and haemophilia]. / Rückenschmerzen und Hämophilie.

A literature research for back pain in hemophilia (1990-2007) revealed only five papers! They all had lumbar or sciatic pain due to hematoma. All symptoms responded to factor VIII replacement. A similar research for a normal population showed hundreds of papers with a lifetime prevalence of 80% for back pain. A survey of 49 patient with hemophilia showed similar results. 70% had experienced back pain before. The reported pain of 40 to 70 on a visual analog scale was significant. 40% reported that the back pain would be more limiting than the pain associated with hemophilia. The hemophilic patient has learned to cope! The treatment of back pain will be of growing importance for hemophilia centers while the typical complaints of hemophilic symptoms will decrease due to better treatment protocols.

Dietary intake and sources of long-chain n-3 PUFAs in German adults.

Dietary intake of long-chain polyunsaturated fatty acids (EPA and DHA) was analysed, stratified by sex and age groups, using data from the German Nutrition Survey 1998. The median intake of both fatty acids combined (EPA and DHA) was 141 mg per day among women and 186 mg among men. In all age groups, women consumed less EPA and DHA than men, partly because of lower total intake. The lowest median intake was observed among women aged 18-24 years (84 mg) and the highest median intake among men aged 45-54 years (217 mg). The main sources of these fatty acids are fish (68%), eggs (12%), poultry (7%), meat and sausages (7%). The remaining 6% of EPA and DHA is supplied by bakery products.

[Telemedical monitoring of electrocardiograms after cardiac surgery--results from a pilot study among 208 patients]. / Telemedizinisches EKG-Monitoring nach herzchirurgischen Operationen: Ergebnisse einer Pilotstudie mit 208 Patienten.

BACKGROUND: An increasing number of older patients undergo cardiac surgery. Complications after cardiac surgery such as rhythm disorders, myocardial infarction and atypical symptoms frequently lead to ambulatory visits and hospitalisations. Telemonitoring might be one method for rapid and efficient detection and classification of symptoms. We examined in this pilot study if ECG-telemonitoring after cardiac surgery proves to be an useful, reliable and accepted procedure with respect to cost and risk reduction. METHODS & RESULTS: Two hundred eight patients (46 female, 162 male) received after surgery an individually adjusted portable 12-lead ECG-monitor. Within three months all incoming ECG-recordings were analysed. In total, 1387 calls from 165 patients (80% use) with ECG-recording (8,4 calls per patient) were collected. There were 235 calls (17%) because of symptoms, 51% of them were registered between 6 PM and 8 AM. Fourteen (6%) out of those 235 emergency calls with ECG-registration led to hospitalisation (n=5) or ambulatory visits next day. The remaining 221 ECG showed no pathological ECG-signs and the patients could be managed with telephonic advice, reassurance and telemedical follow up. Readmissions were due to angina pectoris, severe but unspecific chest pain and cardiac decompensation (n=3) as well as rhythm disturbances (n=2). Almost 75% of all emergency calls were recorded within the first 60 min after the onset of symptoms. CONCLUSION: Older patients reproducibly are able to telemetrically transmit electrocardiograms after a short training before discharged home. Although there is a low incidence of complications among our study population, telemedical ECG-monitoring rapidly helps to differentiate between the symptoms leading to increased patient safety and prevented further damage. The reduction of ambulatory visits and hospitalisations only for treatment of objectified symptoms may lead to a overall cost reduction in the health care system. The reduction of unnecessary hospitalisations and ambulatory visits might also contribute to an optimised time management.

Angiotensin II induces LOX-1, the human endothelial receptor for oxidized low-density lipoprotein.

BACKGROUND: Oxidatively modified LDL (oxLDL) plays an important role in the development of atherosclerosis. OxLDL effects, eg, foam cell formation, are mediated in part by the classic scavenger receptor, whereas other effects may involve the recently cloned endothelial oxLDL receptor, LOX-1 (lectinlike oxLDL receptor-1), which is distinct from macrophage scavenger receptors. Because the regulation of LOX-1 must still be defined, we investigated whether LOX-1 is regulated by the potentially proatherosclerotic stimulant angiotensin II (Ang II). METHODS AND RESULTS: Using competitive reverse transcription-polymerase chain reaction (RT-PCR), we quantified mRNA expression of LOX-1 in primary cultures of human umbilical vein endothelial cells (HUVECs). After treatment with Ang II for 3 hours (1 nmol/L to 1 micromol/L), LOX-1 mRNA was concentration-dependently induced (from 6.9+/-1.4 to 23.1+/-5.5 relative units [RU] by 1 micromol/L Ang II; P<0.05). The angiotensin II type 1 (AT(1)) receptor antagonist losartan prevented this induction. Incubation of HUVECs with Ang II (100 nmol/L, 3 hours) induced LOX-1 protein expression (212+/-21% of control level; P<0. 01) and uptake of 1,1'-dioctadecyl-3,3,3', 3'-tetramethylindocarbocyanine perchlorate (DiI)-labeled oxLDL (209+/-17% of control level; P<0.05) by an AT(1)-dependent pathway, reaching its maximum after 24 hours (680+/-89%; P<0.05). In internal mammary artery biopsy samples from patients with or without ACE inhibitor treatment before coronary artery bypass surgery, LOX-1 mRNA was downregulated by ACE inhibition (6.4+/-2.0 versus 19.3+/-5. 9 RU; n=12 each; P<0.05). CONCLUSIONS: We conclude that LOX-1 is regulated by Ang II in vitro and in vivo, that induction of LOX-1 is mediated by the AT(1) receptor, and that repression of LOX-1 by long-term ACE inhibitor treatment may contribute to the antiatherosclerotic potential of this therapy.

Immunoglobulin V regions and epitope mapping of a murine monoclonal antibody against p24 core protein of HIV-1.

The nucleotide sequence of a murine monoclonal antibody (CB-mab-p24/13-5) against p24 core protein of the human immunodeficiency virus (HIV-1) was determined for variable regions of the heavy and light chain, respectively. Genetic elements encoding the VDJH- and VJL-regions of the antibody were generated from RNA by the polymerase chain reaction, cloned into the vector pICEM 19R and sequenced. Synthetic peptides, 10 amino acids overlapping served for the localization of the epitope. The residues 152-156 within the p24 sequence contain the epitope.

Introduction of the octanucleotide restriction site SwaI into the bicistronic vector pTiSDT for high level synthesis of proteins.

The octanucleotide recognition site for the endonuclease SwaI was introduced into the Escherichia coli bicistronic expression vector pTiSDT by mutating a single position in the coupling SD sequence between a truncated form of the cro-gene and the multicloning site. This mutation does not influence the expression rate. The introduction of this restriction site allows high level production of proteins, that are modified only by an N-terminal methionine incorporated as the start codon.

Transport and fate of gaseous pollutants associated with the National Energy Program.

The experimental evidence related to the long-range transport and transformations of sulfates has been critically reviewed. It has been concluded that sulfate emissions from various sources can be superimposed on each other during long-range transport to create episodes far from the contributing sources. Sulfates are important contributors to degrading visibility through the United States including relatively pristine areas in the western United States. Acid sulfate components, along with nitric acid, may increase the problem of acidity in precipitation on soils, vegetation, and lakes in certain areas of the eastern United States. Improvements in monitoring of these pollutants are required to follow changes in air quality caused by long-range transport.

Expression of monovalent fragments derived from a human IgM autoantibody in E. coli. The input of the somatically mutated CDR1/CDR2 and of the CDR3 into antigen binding specificity.

A hybridoma producing a polyspecific human monoclonal IgM antibody (named CB03) has been derived from a fusion of mouse myeloma cells with human spleen lymphocytes obtained from an autoimmune patient suffering from chronic idiopathic thrombocytopenia. The antibody was found to be encoded by somatically mutated VHI and VlambdaIII genes. To study the input of mutated complementarity regions (CDRs) into antibody specificity, the antigen binding features of the purified complete IgM antibody were compared with (i) a Fab fragment by hot tryptic digestion and (ii) recombinant monovalent fragments expressed in E. coli. In detail, vectors were constructed encoding for (i) rFab03 and single chain Fv03 fragments containing the VH and VL genes connected by a linker sequence, (ii) scFc1.1. fragments containing the VH germline equivalent and the CB03 wild-type CDR3 region, and (iii) scFv fragments containing the CDR1 and CDR2 in germline configuration and the CDR3 expressed in the CB253 human fetal B cell hybridoma producing a polyspecific IgM antibody. The expression vectors contained at the 3' end either a (His)6 motif allowing purification on Ni(2+)-agarose or a c-myc tag for specifically detecting the expression products by a murine monoclonal antibody. Western blotting and ELISA analyses of the expression products indicate: (i) recombinant Fab fragments were found in the bacterial periplasm in extremely low amounts (1-10 micrograms from 1 litre bacterial culture), (ii) scFv fragments were obtained in suitable amounts from bacterial periplasm (800-1000 micrograms/l), (iii) the monovalent recombinant fragments as well as the Fab obtained by tryptic digestion reflected the polyspecific antigen binding features of the complete IgM antibody, but did bind to the antigens with much lower affinity, and (iv) the CDR3 was found to be of critical importance for the antigen binding pattern of this particular IgM. We discuss the expression of recombinant scFv fragments in E. coli as a suitable method in studying the role of the somatic mutation in autoantibody generation.

X ray microscopy--state of the art and expected developments.

The Göttingen X ray microscope at the electron storage ring BESSY in Berlin and X ray microscopy experiments with biological specimens are described. A look ahead to future developments--optics for higher resolution and with better efficiency, detector devices with higher detective quantum efficiency, and the development of a laboratory X ray microscope with a plasma X ray source--is taken.

Computed tomography of cryogenic biological specimens based on X-ray microscopic images.

Soft X-ray microscopy employs the photoelectric absorption contrast between water and protein in the 2.34-4.38 nm wavelength region to visualize protein structures down to 30 nm size without any staining methods. Due to the large depth of focus of the Fresnel zone plates used as X-ray objectives, computed tomography based on the X-ray microscopic images can be used to reconstruct the local linear absorption coefficient inside the three-dimensional specimen volume. High-resolution X-ray images require a high specimen radiation dose, and a series of images taken at different viewing angles is needed for computed tomography. Therefore, cryo microscopy is necessary to preserve the structural integrity of hydrated biological specimens during image acquisition. The cryo transmission X-ray microscope at the electron storage ring BESSY I (Berlin) was used to obtain a tilt series of images of the frozen-hydrated green alga Chlamydomonas reinhardtii. The living specimens were inserted into borosilicate glass capillaries and, in this first experiment, rapidly cooled by plunging into liquid nitrogen. The capillary specimen holders allow image acquisition over the full angular range of 180 degrees. The reconstruction shows for the first time details down to 60 nm size inside a frozen-hydrated biological specimen and conveys a clear impression of the internal structures. This technique is expected to be applicable to a wide range of biological specimens, such as the cell nucleus. It offers the possibility of imaging the three-dimensional structure of hydrated biological specimens close to their natural living state.