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Hemoglobin microparticles (HbMP) produced with a three-step procedure including co-precipitation of hemoglobin with manganese carbonate, protein crosslinking and dissolution of the carbonate template were shown to be suitable for application as artificial oxygen carriers. First preclinical safety investigations delivered promising results. Bacterial safety plays a decisive role during the production of the HbMP. Therefore, bioburden and endotoxin content of the starting materials (especially hemoglobin) and the final particle suspension are intensively tested. However, some bacteria may not have been detected by the standard tests due to low concentration. The aim of this study was to investigate how these bacteria would behave in the fabrication process. Biocidal effects are known for glutaraldehyde and for ethylenediaminetetraacetic acid, chemicals that are used in the fabrication process of HbMP. It could be shown that both chemicals prevent bacterial growth at the concentrations used during the HbMP fabrication. In addition, the particle production was carried out with hemoglobin solutions spiked with Escherichia coli or Staphylococcus epidermidis. No living bacteria could be detected in the final particle suspensions. Therefore, we conclude that the HbMP fabrication procedure is safe in respect of a bacterial contamination.

Background: Although transmission of pathogenic viruses through human tissue grafts is rare, it is still one of the most serious dreaded risks of transplantation. Therefore, in addition to the detailed medical and social history, a comprehensive serologic and molecular screening of the tissue donors for relevant viral markers for human immunodeficiency virus (HIV), hepatitis B virus (HBV), and hepatitis C virus (HCV) is necessary. In the case of reactive results in particular, clear decisions regarding follow-up testing and the criteria for tissue release must be made. Methods: Based on the clinical relevance of the specific virus markers, the sensitivity of the serological and molecular biological methods used and the application of inactivation methods, algorithms for tissue release are suggested. Results: Compliance with the preanalytical requirements and assessment of a possible hemodilution are mandatory requirements before testing the blood samples. While HIV testing follows defined algorithms, the procedures for HBV and HCV diagnostics are under discussion. Screening and decisions for HBV are often not as simple, e.g., due to cases of occult HBV infection, false-positive anti-HBc results, or early window period positive HBV NAT results. In the case of HCV diagnostics, modern therapies with direct-acting antivirals, which are often associated with successful treatment of the infection, should be included in the decision. Conclusion: In HBV and HCV testing, a high-sensitivity virus genome test should play a central role in diagnostics, especially in the case of equivocal serology, and it should be the basis for the decision to release the tissue. The proposed test algorithms and decisions are also based on current European recommendations and standards for safety and quality assurance in tissue and cell banking.

Background Galactosylation of immunoglobulin G (IgG) is reduced in rheumatoid arthritis (RA) and assumed to correlate with inflammation and altered humoral immunity. IgG hypogalactosylation also increases with age. To investigate dependencies in more detail, we compared IgG hypogalactosylation between patients with RA, patients with axial spondyloarthritis (axSpA), and healthy control subjects (HC), and we studied it in RA on the background of HLA-DRB1 shared epitope (SE), anticitrullinated protein antibodies (ACPA), and/or rheumatoid factor (RF) status. Methods Patients with RA (n = 178), patients with axSpA (n = 126), and HC (n = 119) were characterized clinically, and serum IgG galactosylation was determined by capillary electrophoresis. Markers of disease activity, genetic susceptibility, and serologic response included C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), DAS28, SE, HLA-B27, ACPA, and RF. Expression of glycosylation enzymes, including beta 1–4 galactosyltransferase (B4GALT3) activity, were estimated from transcriptome data for B-cell development (GSE19599) and differentiation to plasma cells (GSE12366). Results IgG hypogalactosylation was restricted to RA and associated with increasing CRP levels (p < 0.0001). In axSpA, IgG hypogalactosylation was comparable to HC and only marginally increased upon elevated CRP. Restriction to RA was maintained after correction for CRP and age. Treatment with sulfasalazine resulted in significantly reduced IgG hypogalactosylation (p = 0.003) even after adjusting for age, sex, and CRP (p = 0.009). SE-negative/ACPA-negative RA exhibited significantly less IgG hypogalactosylation than all other strata (vs SE-negative/ACPA-positive, p = 0.009; vs SE-positive/ACPA-negative, p = 0.04; vs SE-positive/ACPA-positive, p < 0.02); however, this indicated a trend only after Bonferroni correction for multiple testing. In SE-positive/ACPA-negative RA IgG hypogalactosylation was comparable to ACPA-positive subsets. The relationship between IgG hypogalactosylation and disease activity was significantly different between strata defined by SE (CRP, p = 0.0003, pBonferroni = 0.0036) and RF (CRP, p < 0.0001, pBonferroni < 0.0012), whereas ACPA strata revealed only a nonsignificant trend (p = 0.15). Gene expression data indicated that the key enzyme for galactosylation of immunoglobulins, B4GALT3, is expressed at lower levels in B cells than in plasma cells. Conclusions Increased IgG hypogalactosylation in RA but not in axSpA points to humoral immune response as a precondition. Reduced B4GALT3 expression in B cells compared with plasma cells supports relatedness to early B-cell triggering. The differential influence of RA treatment on IgG hypogalactosylation renders it a potential diagnostic target for further studies. Electronic supplementary material The online version of this article (10.1186/s13075-018-1540-0) contains supplementary material, which is available to authorized users.

BACKGROUND: Hypertension is one of the leading global risks for cardiovascular events worldwide. There is preliminary evidence that regular blood donation may be beneficial. STUDY DESIGN AND METHODS: Unselected blood donors were included in this observational study. Blood pressure (BP) was measured before and after blood donation, with participants donating between one and four occasions in a 1-year study period. RESULTS: In this study, 292 donors were enrolled. At baseline, 146 had elevated BP (>140/90 mmHg). In hypertensives, after four blood donations, systolic and diastolic blood pressure (SBP and DBP, respectively) decreased from a mean of 155.9 613.0 to 143.7 615.0 mmHg and from 91.4 69.2 to 84.5 69.3 mmHg, respectively (each p

The poor healing potential of tendons is still a clinical problem, and the use of Platelet Rich Plasma (PRP) was hypothesized to stimulate healing. As the efficacy of PRPs remains unproven, platelet lysate (PL) could be an alternative with its main advantages of storage and characterization before use. Five different blood products were prepared from 16 male donors: human serum, two PRPs (Arthrex, (PRP-ACP); RegenLab (PRP-BCT)), platelet concentrate (apheresis, PC), and PL (freezing-thawing destruction of PC). Additionally, ten commercial allogenic PLs (AlloPL) from pooled donors were tested. The highest concentration of most growth factors was found in AlloPL, whereas the release of growth factors lasted longer in the other products. PRP-ACP, PRP- BCT, and PC significantly increased cell viability of human tenocyte-like cells, whereas PC and AlloPL increased Col1A1 expression and PRP-BCT increased Col3A1 expression. MMP-1, IL-1β, and HGF expression was significantly increased and Scleraxis expression decreased by most blood products. COX1 expression significantly decreased by PC and AlloPL. No clear positive effects on tendon cell biology could be shown, which might partially explain the weak outcome results in clinical practice. Pooled PL seemed to have the most beneficial effects and might be the future in using blood products for tendon tissue regeneration.

Acquired hemophilia A (AHA) is caused by autoantibodies against factor VIII (FVIII). Immunosuppressive treatment (IST) results in remission of disease in 60% to 80% of patients over a period of days to months. IST is associated with frequent adverse events, including infections as a leading cause of death. Predictors of time to remission could help guide IST intensity but have not been established. We analyzed prognostic factors in 102 prospectively enrolled patients treated with a uniform IST protocol. Partial remission (PR; defined as no active bleeding, FVIII restored >50 IU/dL, hemostatic treatment stopped >24 hours) was achieved by 83% of patients after a median of 31 days (range 7-362). Patients with baseline FVIII

The analytical performance of the Veris HIV-1 assay for use on the new, fully automated Beckman Coulter DxN Veris molecular diagnostics system was evaluated at 10 European virology laboratories. The precision, analytical sensitivity, performance with negative samples, linearity, and performance with HIV-1 groups/subtypes were evaluated. The precision for the 1-ml assay showed a standard deviation (SD) of 0.14 log 10 copies/ml or less and a coefficient of variation (CV) of ≤6.1% for each level tested. The 0.175-ml assay showed an SD of 0.17 log 10 copies/ml or less and a CV of ≤5.2% for each level tested. The analytical sensitivities determined by probit analysis were 19.3 copies/ml for the 1-ml assay and 126 copies/ml for the 0.175-ml assay. The performance with 1,357 negative samples demonstrated 99.2% with not detected results. Linearity using patient samples was shown from 1.54 to 6.93 log 10 copies/ml. The assay performed well, detecting and showing linearity with all HIV-1 genotypes tested. The Veris HIV-1 assay demonstrated analytical performance comparable to that of currently marketed HIV-1 assays. (DxN Veris products are Conformité Européenne [CE]-marked in vitro diagnostic products. The DxN Veris product line has not been submitted to the U.S. FDA and is not available in the U.S. market. The DxN Veris molecular diagnostics system is also known as the Veris MDx molecular diagnostics system and the Veris MDx system.)

SummaryBackground: Infectious disease marker testing is obligatory for the release of human tissue for transplantation. Most CE-marked tests are not validated for postmortem blood. In a previous study we have validated the testing for anti-HIV-1/2, anti-HCV, HBsAg, and anti-HBc. Here, we present the validation of testing for antibodies against T. pallidum, which is the last marker obligatory for tissue release for transplantation. Methods: 17 samples of postmortem sera and 10 samples of both pre- und postmortem sera were obtained from cornea donors and tested for anti-T. pallidum on the Siemens-BEP-III-System. These sera were spiked with anti-T. pallidum-positive standard sera in concentrations which give low- and high-positive results at the respective dilution. Results: Two of the unspiked postmortem sera were false-positive most likely due to intense hemolysis (free hemoglobin > 50 mg/dl). Of the 25 negative postmortem sera, none of the spiked samples was false-negative after 0, 24 and 60 h. Conclusion: There is no indication that postmortem samples give false-negative or false-positive results with the test system and test kits used in cases of low hemolysis. The procedure described might serve as a model for validating other test kits on postmortem samples.

Defined serological blood tests of deceased cornea donors are required to minimize the risk of viral infections of a transplant recipient as much as possible. Haemolysis, autolysis and bacterial contamination, may produce significant changes of post-mortem blood samples, which may lead to false serological test results. Pre- and post-mortem findings from the same cornea donors of the University Tissue Bank of the Charite in the years 2004–2009 (n = 487) were retrospectively analyzed and compared. The test results from pre-mortem blood samples were defined as the reference for the post-mortem blood test. Of 487 cornea donors, there were a total of 21 cases (4.3 %) with discrepancies between serological test results from pre- and post-mortem blood samples. Of these, 7 values referred to the HBsAg-testing, 3 to the anti-HBs-, 1 to the anti-HBcIgG + IgM-, 1 to the anti-HCV-, 4 to the anti-HIV 1/2- and 5 to the TPLA-findings. False negative results within post-mortem serology occurred in 4 of 487 cases (0.8 %). False positive results within the post-mortem blood samples occurred at a much more frequent rate, with 17 of 487 cases (3.5 %). Discrepancies between serological pre- and post-mortem blood tests occur mainly due to the use of non-validated test systems. Therefore, it seems reasonable to test pre- and post-mortem blood samples serologically, whenever possible, at the same time, regardless of the sample age. Positive results, regardless of the sample type, should always be retested with validated confirmation tests (e.g. NAT), in order to differentiate between false and true positive results.

Objective According to EU regulations (EU directive 2006/17/EC), blood specimens for virologic testing in the context of post-mortal tissue donation must be taken not later than 24 h post mortem.

The impact of epigenetics on the differentiation of memory T (Tmem) cells is poorly defined. We generated deep epigenomes comprising genome-wide profiles of DNA methylation, histone modifications, DNA accessibility, and coding and non-coding RNA expression in naive, central-, effector-, and terminally differentiated CD45RA

Red vine leaf extract (RVLE) is a herbal medicine containing several flavonoids, with quercetin-3-O-s-glucuronide and isoquercitrin (quercetin-3-O-s-glycoside) as the main components. Objective – To assess the efficacy and safety of once-daily doses of 360 and 720 mg RVLE (pharmaceutical extract code AS 195; Antistax® Venenkapseln) compared to placebo in patients with stage I and incipient stage II chronic venous insufficiency (CVI). Design – A 12-week, randomized, double-blind, placebo-controlled, parallel-group, multi-center study. Patients – Male and female outpatients aged 25 to 75 years with stage I to stage II CVI (i.e. without extensive trophic changes), not having any other significant medical conditions and not treated with compression stockings, diuretics or other drugs affecting fluid balance. Intervention – Patients were randomly assigned to a double-blind treatment with placebo, 360 mg AS 195 or 720 mg AS 195 once daily for 12 weeks, preceded and followed by a single-blind 2-week placebo treatment for baseline run-in and end-of-trial washout, respectively. Study criteria were evaluated at baseline, after 6 and 12 weeks of treatment and 2 weeks after discontinuation of treatment. Results – Of the 260 patients enrolled and randomized, 219 completed the study in accordance with the protocol. In the intention-to-treat analysis (N = 257), the mean (± SD) lower leg volume (measured by water displacement plethysmography) of the patients treated with placebo (N = 87) increased by 15.2 ± 90.1 g (displaced water mass) and by 33.7 ± 96.1 g after 6 and 12 weeks compared to baseline. In contrast, for patients treated with AS 195, lower leg volume decreased, and after 12 weeks of treatment, the difference in mean lower leg volume between the active treatment groups and the placebo group was −75.9 g (95% CI: −106.1 to −45.8 g) and −99.9 g (95% CI: −130.3 to −69.6 g) for the group treated with 360-mg AS 195 (N = 86) and 720-mg AS 195 (N = 84), respectively. The changes in calf circumference showed a similar pattern: in patients treated with AS 195, both the higher dose (720 mg) and, albeit to a lesser extent, the lower dose (360 mg) resulted in a clear reduction in circumference over time, whereas, circumferences remained largely unchanged in patients treated with the placebo (95% CI of the estimated treatment effects vs. placebo after 12 weeks: -1.40 to -0.56 cm and -1.73 to -0.88 cm for 360 and 720 mg AS 195, respectively). These differences were statistically significant (p Conclusion – Once-daily doses of 360 and 720 mg AS 195 were confirmed to be safe and effective in the treatment of mild CVI, reducing significantly lower leg edema and circumference whilst improving key CVI-related symptoms to a clinically relevant extent. The edema reduction is at least equivalent to that reported for compression stockings and/or other edema-reducing agents. The higher dose was as well tolerated as the lower dose but resulted in a slightly greater and more sustained improvement.

Background: Diagnosis of giant cell arteritis (GCA) is based on criteria of the American College of Rheumatology. However, not all GCA patients meet these criteria and treatment may be delayed in individual patients, leading to an increased risk of complications. Methods: In an observational study, we investigated acute phase response markers in GCA and non-GCA patients matched for erythrocyte sedimentation rate and CRP levels. Results: Plasma viscosity (PV) was significantly elevated in all GCA patients, but normal in non-GCA patients. Conclusions: Our data suggest that PV may reflect a more specific component of the acute inflammatory response in patients with GCA. Analysis of PV may significantly contribute to a reliable diagnosis early in the course of the disease, particularly in patients with suspected GCA that do not meet current diagnostic criteria.

Summary Background: Some properties of blood are modified post mortem. These modifications might give false-negative or false-positive results in infectious disease testing. Most CE-marked test equipment for infectious serology testing is not validated for testing post-mortal blood. Validation, however, is obligatory, if the results are used for the release of tissues for transplantation. Methods: Samples of pre- and post-mortem sera were obtained from 20 cornea donors, and the results were compared for anti-HIV-1/2, anti-HCV, HBsAg, and anti-HBc on the Siemens-BEP-III Automatic System. Negative post-mortem sera were spiked with standard sera (PEI anti-HCV IgG, PEI HBsAg ad 1000 standard, anti-HBc IgG (WHO) NIBSC 95/522, PEI anti-HIVIV) in concentrations which give low- and high-positive results for the respective marker. Results: All pre-mortem sera were negative for all markers. None of the post-mortem samples was false-positive. None of the spiked postmortem samples was false-negative. Technical errors occurred during the validation process but could be detected and eliminated. Serum samples should be centrifuged immediately after collection, and it must be taken into account that post-mortem serum could rarely lead to blockage of pipetting systems due to clotting phenomena. Conclusion: There is no indication that post-mortem samples give false-negative or false-positve results with the test system and test kits used. The procedure described might serve as a model for validating other test kits on post-mortem samples.

BACKGROUND: The FREELYS Nano system (Diagast) is a manual workstation for ABO/D grouping, Rh phenotyping, K typing, and antibody screening (ABS) for immunoglobulin G (IgG) antibodies only and works with the erythrocyte-magnetized technology (EMT). The principle of EMT is based on magnetization of red blood cells and avoids centrifugation and washing steps. STUDY DESIGN AND METHODS: A total of 304 samples were tested with our routine blood bank methods, 100 samples for ABO/D grouping, 196 samples (100 at first evaluation, 96 at second evaluation) for Rh phenotyping and K typing (PK7200, Olympus), and 108 samples for ABS (DiaMed). All samples were tested in parallel with the FREELYS Nano. RESULTS: We found a 100% concordance between the observed (FREELYS Nano) and the expected (Olympus PK7200) results for ABO/D grouping in all 100 samples. For Rh phenotyping and K tests, in 24 of 100 samples false-positive reactions were observed in the first evaluation by the FREELYS Nano. After changing the test kit batch for Rh phenotyping by the manufacturer, a complete concordance in Rh phenotyping and K tests was observed in a second evaluation. For ABS, the FREELYS Nano showed in 4 of 108 samples (3.7%) false-negative reactions for IgG antibodies (two anti-K, one anti-E, one anti-Cw), and one (0.9%) false-positive reaction. CONCLUSIONS: The FREELYS Nano is reliably suited to ABO/D grouping, Rh phenotyping, and K testing. The rate of false-negative reactions for IgG antibodies should be reduced.

In this prospective, randomized, placebo-controlled clinical study we aimed to investigate the clinical effect of a Cistus extract (CYSTUS052) in 160 patients with infections of the upper respiratory tract. The extract contains a high percentage of highly polymeric polyphenols. In cell culture and in a mouse model it exerts antiviral and antimicrobial activities. Principal active constituents of the genus Cistus are polyphenolic compounds. Plant-derived polyphenols have been shown to be strong antioxidants with potential health benefits. Various reports have appeared on the antiviral and antibacterial potential, including several reports describing the antiviral activity of polyphenols against influenza virus. Clinical studies on the effectiveness of Cistus incanus are scarce. Only one controlled application observation study demonstrated the effectiveness of a Cistus extract. The present randomised, placebo-controlled clinical study was designed to compare the symptom scores in patients with common cold treated either with CYSTUS052 or with placebo. A score of subjective symptoms decreased significantly over the course of treatment with Cistus, whereas treatment with placebo resulted in a less distinct decrease of symptoms. Among the inflammatory markers investigated, the C-reactive protein was mostly affected by Cistus and decreased significantly in the treatment group.

BACKGROUND: Human immunodeficiency virus Type 1 (HIV-1) assays applying nucleic acid testing (NAT) rely on HIV-1 sequence-specific primers and probes. Their hybridization can be limited or abolished by genetic polymorphisms occurring in the target sequence. STUDY DESIGN AND METHODS: Blood donations are routinely tested for HIV-1/2 antibodies and for HIV-1 RNA in our blood transfusion unit. Recently, HIV-1 RNA was undetectable with an established in-house real-time long terminal repeat (LTR) reverse transcriptase–polymerase chain reaction (RT-PCR) in two cases, whereas serologic assays were positive. The reason for this discrepancy was elucidated by sequencing of the NAT target region in the respective single donations. An improved primer was designed and tested on HIV-1 reference panels and blood donations to ensure reliable detection of HIV-1 RNA. RESULTS: Direct sequencing of the target region, isolated from samples of two unrelated HIV-positive blood donors, revealed one and four mismatches in the hybridization domain of the forward primer, respectively. Both viruses belong to HIV-1 Subtype B. LTR RT-PCR with an additional forward primer was suitable for all strains of HIV-1 tested with high sensitivity. CONCLUSIONS: Surveillance of HIV-1 genetic diversity is essentially required to continually evaluate its impact on performance of diagnostic and patient monitoring assays.

Examples of medicinal herbs that have been perpetuated along several generations based simply on a folk tradition are Cistus and green tea. The principal active constituents of the genus Cistus and green tea are polyphenolic compounds. Polyphenols exhibit a wide range of antibacterial, antifungal and antiinflammatory effects. The present work aimed to investigate the clinical effect of a Cistus extract (CYSTUS052) in comparison with green tea on 300 patients with infections of the upper respiratory tract. Due to the lack of clinical study data on their efficacy in patients, this is a report of the findings of our study on the clinical efficacy of CYSTUS052 in patients with the upper respiratory tract infections (URTIs). This study observed a total of 300 patients (277 completers) treated with CYSTUS052 given in lozenges compared with treatment with an extract of green tea. The patients scored the subjective severity of target symptoms using a predefined scale. The score of subjective symptoms decreased over the course of treatment with CYSTUS052, whereas treatment with green tea resulted in a less significant decrease of symptoms. CYSTUS052 therefore proved to be an effective adjuvant for the treatment of respiratory infections.

BACKGROUND: It is unknown whether human immunodeficiency virus (HIV) can be transmitted via blood products donated after primary HIV-1 infection before the detection of viral RNA in plasma. CASE REPORT: From a 39-year-old repeat donor, double plateletpheresis donations were collected on Days 4 and 18 after the presumptive date of primary HIV-1 infection. The former apheresis donations tested negative for the presence of HIV and were transfused to two patients, whereas the latter donation tested positive by reverse transcription–polymerase chain reaction (RT-PCR) but not in antibody screening and was not released for transfusion. RESULTS: One of the recipients of the Day 4 apheresis donation died of unrelated reasons and could therefore not be tested. The second recipient did not develop HIV-1 infection and has remained negative for the presence of all HIV markers over a period of 7.5 months after the receipt of the apheresis unit. In the donor, qualitative and quantitative RT-PCR as well as an antibody-antigen combination assay were observed to be positive on Day 18. In contrast, the HIV antibody screening test became positive for the first time on Day 25. CONCLUSION: Transmission of HIV-1 may not occur during the very early stage of infection.

Mit der Umsetzung der EU-Richtlinien 2004/23/EG und 2006/17/EG im Gewebegesetz sowie den begleitenden Verordnungen (Arzneimittel- und Wirkstoffherstellungsverordnung, Transplantationsgesetz-Gewebeverordnung) wurden die grundlegenden Anforderungen an die Virussicherheit der Gewebespenden allgemein definiert. Wahrend infektionsserologische Testungen (Anti-HIV 1/2, Anti-HCV, Hepatitis-B-Oberflachenantigen, Anti-Hepatitis-B-Core-Antigen, Treponema-pallidum-Hamagglutinationsassay) vorgeschrieben sind, wird der Nukleinsaure-Nachweis fur HIV, HBV und HCV nicht explizit gefordert. Anhand in der Literatur berichteter Virusubertragungen, gewebespezifischer Besonderheiten sowie Herstellungs- und gegebenenfalls Inaktivierungsverfahren wird eine Bewertung des Stellenwertes des HIV/HCV/HBV-Nachweises mittels Nukleinsaure-Amplifikationstechniken (NAT) bei Spendern unterschiedlicher Gewebe vorgenommen und mit den Erfahrungen des Blutspendewesens verglichen. Muskuloskelettale Gewebe besitzen infolge des zumeist hohen Blutgehalts der Gewebe, des umfangreichen Entnahmespektrums, der bisher beschriebenen Ubertragungen, der unterschiedlichen Herstellungsverfahren sowie der hohen Spender-Empfanger-Ratio ein signifikantes Risiko, HIV/HCV/HBV zu ubertragen. Daher sollte bei Spendern muskuloskelettaler Gewebe, die keinem effektiven Virusinaktivierungsverfahren unterzogen werden, eine HIV-, HBV- und HCV-NAT-Testung erfolgen. Kardiovaskulares Gewebe hat bei Durchfuhrung der serologischen Testung ein sehr geringes Restrisiko der HIV/HCV/HBV-Ubertragung. Aufgrund der fehlenden Moglichkeit einer effektiven Virusinaktivierung (Erhalt der Gewebemorphologie) und der Spender-Empfanger-Ratio von bis zu 1:10 sollte die HIV-, HCV- und HBV-NAT jedoch als zusatzliche Sicherheitsmasnahme erfolgen. Augenhornhaute besitzen aus physiologisch-morphologischer sowie epidemiologischer Sicht das geringste HIV/HCV/HBV-Ubertragungsrisiko, jedoch sollte die HCV-NAT durchgefuhrt werden. Eine Quarantanelagerung der Gewebe eines Spenders kann bei negativem Ergebnis der HIV/HCV/HBV-NAT grundsatzlich entfallen. Aufgrund der vielfaltigen Synergieeffekte mit der Transfusionsmedizin bietet es sich fur Gewebebanken an, die Testung der infektionsserologischen bzw. molekularbiologischen Laborparameter in Kooperation mit Blutspendediensten durchzufuhren.

SummaryPatients with refractory autoimmune thrombocytopenia (ITP) may develop life-threatening bleeding that cannot be immediately controlled by drug administration. To date, there have been no studies conducted to evaluate the efficacy of platelet transfusion alone in such cases. Ten patients with refractory ITP and bleeding or a high bleeding risk were consecutively transfused (one unit/30 min) with apheresis platelet concentrates (APC) without the administration of new drugs. The used APCs (average 3 – 7 units) contained 2.7 × 1011 (medium) platelets and were leukodepleted (≤1 × 106 leukocytes/unit).Platelet serology was performed using standard techniques. Platelet transfusion resulted in an increase in the platelet count to 84 – 157 × 103/µl, and the cessation of bleeding in all patients without any serious adverse effects. Although platelet counts gradually decreased within a few days post-transfusion, bleeding was stopped in all cases. These findings indicate that consecutive platelet transfusion using APCs is a rapidly effective emergency treatment in patients with refractory ITP.

Hintergrund: Die Anwendung allogener Gewebetransplantate wie Augenhornhaut, Herzklappen und Gefase sowie muskuloskelettale Gewebe ist fester Bestandteil der Therapie von Gewebedefekten. Die Gewinnung dieser Gewebe kann nach standardisierten Verfahren im Rahmen einer Organspende erfolgen, bei denen auch die Einwilligung zur Gewebeentnahme vorliegt. Methoden: In der Region Nord-Ost (Berlin, Brandenburg, Mecklenburg-Vorpommern) der Deutschen Stiftung Organtransplantation (DSO) wurde bereits in den 1990er Jahren ein Gewebespendeprogramm bei Organspendern entwickelt. Ergebnisse: Durch die Kooperation von Spendekrankenhausern, der DSO/DSO-G (Gemeinnutzige Gesellschaft fur Gewebetransplantation) und ortsansassigen Gewebebanken war es im Zeitraum der Jahre 2003–2006 moglich, bei 71,4% (433/609) der effektiven Organspender Augenhornhaut und bei 70,3% (428/609) muskuloskelettale Gewebe fur Transplantationszwecke zu entnehmen. Die Einwilligung zur Entnahme von Herzen zur Transplantation und zur Herzklappengewinnung lag bei 91,4%. Diskussion: Durch das neue deutsche Gewebe gesetz wurde die Gewebespende und -entnahme dem Transplantations- und dem Arzneimittelgesetz sowie den entsprechenden Verordnungen untergeordnet. Deshalb mussen bisher geltende Verfahrensweisen dem Gesetz angepasst werden, um auch weiterhin zu gewahrleisten, dass die Entnahme und Ubertragung vermittlungspflich tiger Organe stets Vorrang hat, aber auch dem Willen des Verstorbenen bzw. seiner Angehorigen bezuglich der Gewebespende entsprochen wird.

The risk of transmitting human pathogenic viruses via allogeneic musculoskeletal tissue transplants is a problem requiring effective inactivation procedures. Virus safety of bone transplants was achieved using peracetic acid (PAA)-ethanol sterilisation. Proteins are known to have an adverse effect on the virus-inactivating capacity of PAA. Therefore we investigated virus inactivation by PAA in collagenous tissues. Achilles tendon, skin and cartilage were cut into small pieces, lyophilised and contaminated with pseudorabies virus (PRV) or porcine parvovirus (PPV). The inactivating capacity of PAA-ethanol was investigated by determining virus titres in the supernatant or the tissue pellet at different time-points. In all virus-contaminated tissue samples treatment for 10 min with PAA-ethanol resulted in titre reductions by a factor of10(3). PRV was rapidly inactivated below the detection limit (or =2.8 x 10(1) TCID(50)/ml). After 240 min a reduction by a factor of10(4) was obtained for PPV in all samples, but a residual infectivity remained. Collagenous proteins of Achilles tendon, skin and cartilage had no adverse effect on the virus-inactivating capacity of PAA. PAA-ethanol used in the production process at the Charité tissue bank can therefore be recommended for treatment of non-osseous musculoskeletal tissues.

BACKGROUND: In this study, we evaluated the effects of the thawing process of 2 commercially available devices on the activity of clotting factors, inhibitors and activation markers of the hemostatic system in fresh-frozen plasma (FFP). In an experimental procedure, FFP was thawed under running warm water at 42 degrees C. METHODS: Plasma of 20 healthy donors was sampled, separated, and distributed in 3 plasma bags. Within 2 h after sampling plasma bags was frozen at a temperature of -30 degrees C to -40 degrees C and stored for at least 8 wk. After sampling (baseline) as well as immediately and 1, 2, 4, and 6 h after thawing, the activity of FV, FVII, FVIII, fibrinogen, fibrin monomers (FM), d-dimers (DD), alpha2-antiplasmin (alpha2-AP), and protein S (PS) was determined from each plasma bag. RESULTS: From 1 h to 6 h after thawing, no significant differences in the activity of the investigated coagulation markers dependent on the thawing procedure were found. However, immediately after thawing and independent of the thawing procedure, the activity of FVII was significantly decreased (P < 0.01), whereas FM were significantly increased (P = 0.001). CONCLUSION: The thawing procedures studied exhibited no significant influence on activity and stability of the investigated markers of coagulation over the study period. The decreased activity of FVII and the clinical significance of the increase in FM require further research.

Portable photometers are increasingly used for hemoglobin screening in blood centers. In a total of 500 unselected prospective blood donors, venous hemoglobin concentrations were measured using a hematology analyzer, and fingerstick hemoglobin concentrations were measured on both a HemoCue B and a Biotest hemoglobin photometer. In 100 of the donors, earstick hemoglobin levels were also measured on the HemoCue system. The mean hemoglobin levels in the fingerstick samples were slightly higher than those in the venous samples. The deviation exceeded the limit of +/-10g l(-1) in 9% of samples tested by HemoCue and in 20% of those investigated by Biotest. The use of earstick samples led to considerable overestimation of the actual hemoglobin concentration.

The removal of fat from cancellous bone tissue promotes the clinical healing of the transplant and improves the penetration of chemical sterilisation media into the tissue. Treatment using chloroform/methanol (2:1, 2 h) is frequently used as a defatting procedure. Eight rinses with methanol followed by two rinses with aqua ad iniectabilia (20 min each, with ultrasonic effect) ensure depletion in the level of chloroform from defatted cancellous bone to a concentration below 25 ppm (limit value). For the necessary routine quality checks on the production process, a gas chromatography method has been developed that determines the level of chloroform in cancellous bone, for which the detection limit is 0.003 ppm.

Background . Universal leukodepletion of blood components to prevent acute non-hemolytic transfusion reactions (NHTRs) is still a subject of debate. Patients and methods . Transfusion-associated NHTRs observed at our hospital in the last 6 years were retrospectively analyzed. Buffy-coat depleted red blood cells (bc-RBCs), and if indicated, leucodepleted post-storage (ld-RBCs) were initially used. In April 1997, universal leukodepletion was implemented at our hospital, and thereafter only prestorage ld-RBCs were used. All platelet concentrates transfused during this time were prestorage filtered single-donor apheresis platelets (SDAPs). Results . A total of 163,090 blood products were transfused from April 1995 to April 2001 (bc-RBC: n =34,040 units; ld-RBC: n =66,967; SDAP: n =14,516; FFP: n =47,567). The number of post-transfusion febrile NHTRs occurring with each blood product was 65 (0.19%) for bc-RBCs, 8 (0.16%) for post-storage ld-RBCs, 16 (0.03%) for prestorage ld-RBCs, 16 (0.11%) for SDAPs, and 10 (0.02%) for FFP. Allergic reactions ( n =116) were most frequently observed after SDAP transfusion (0.32%) and occurred at a similarly low rate after transfusion of all other blood components (0.03–0.08%). Conclusion . In conclusion, acute NHTRs rarely occur after the use of leukodepleted blood components. Prestorage appears to be more effective than post-storage leukodepletion in preventing febrile reactions following a blood transfusion.

To investigate the effect of the red vine leaf extract AS 195 on cutaneous microvascular blood flow, transcutaneous oxygen pressure (tcpO2), and leg oedema in patients with chronic venous insufficiency (CVI).The study was a randomised, double-blind, placebo-controlled, crossover trial for which 129 men and women, agedor =18 years, with CVI stage I or II were screened. Seventy-one fulfilled the inclusion criteria and were randomised.A total of 71 patients were divided into two groups. The first group (n = 36) received AS 195 360mg once daily during a first 6-week treatment period, followed by a 4-week placebo washout period and then placebo during the second 6-week treatment period. The second group (n = 35) started with placebo and received AS 195 360mg after the placebo washout. The cutaneous microvascular blood flow in the malleolar region was measured using a newly developed laser Doppler device. TcpO2 was measured using a solid-state electrode.After 6 weeks, patients in the AS 195 group had increased microvascular blood flow values (+241.8 +/- 18.7 arbitrary units [AU] versus a decrease of -41.0 +/- 18.7AU in the placebo group; p0.0001). Oxygen increased to 1.35 +/- 0.97mm Hg (placebo: decrease of -7.27 +/- 0.97mm Hg; p0.0001). After 6 weeks of treatment the leg circumference was decreased (ankle level: by -0.39 +/- 0.09cm versus +0.29 +/- 0.09cm; p0.0001; calf level: by -0.54 +/- 0.05cm versus +0.14 +/- 0.05cm; p0.0001).The administration of AS 195 improved objective symptoms of CVI and may prevent CVI deterioration.

Introduction: Disturbance of the uteroplacental circulation (UPC) and the renin-angiotensin system are involved in the pathogenesis of preeclampsia. In women with history of preeclampsia persistently elevated C-reactive protein (CRP) levels have been described. The angiotensin-converting enzyme (ACE) intron 16 insertion/deletion (I/D) genotype is associated with ACE activity and assumed to be a risk factor for preeclampsia. As ACE generates proinflammatory angiotensin II, we analysed, whether ACE intron 16 I/D genotype is associated with CRP and whether this association exhibited a relation to uteroplacental dysfunction. Materials and methods: A total of 639 women have been followed during pregnancy with repeated measurements of CRP levels (observations: n =2333). ACE intron 16 I/D genotype was determined, and its association with CRP was assessed with adjustment for non-independent observations. Results: CRP levels of ACE D allele carriers were significantly higher than those of the ACE II (wild-type) genotype ( p =0.0003, p adj =0.04). This relation was allele-dose dependent ( p

Die Differentialdiagnostik und Therapie von perioperativen Blutungen besitzt in der Gefas- und Kardiochirurgie bei Eingriffen mit hohen Blutverlusten einen besonderen Stellenwert. So ist es sinnvoll, Patienten mit einem erhohten Blutungsrisiko bereits praoperativ zu erfassen und adaquat vorzubereiten. Insbesondere die praoperative Erfassung von Thrombopathien [medikamenteninduzierte Thrombopathien (insbesondere durch ASS), von-Willebrand-Jurgens-Syndrom, andere angeborene Thrombopathien] ist bei der Minimierung von Blutverlusten effektiv. Nicht elektive Situationen (Blutung bei Notoperationen, Akutpatienten) mussen oft ohne gezielte hamostaseologische Diagnostik rational therapiert werden. Einfache Schemata sind fur ein schnelles Handeln unabdingbar. In dieser Arbeit soll versucht werden, Hinweise zu Diagnostik und Therapie der hamorrhagischen Diathese mit den Schwerpunkten: Diagnostik und Therapie der thrombozytar bedingten hamorrhagischen Diathese und Diagnostik und Therapie der plasmatisch bedingten hamorrhagischen Diathese aus hamostaseologisch-transfusionsmedizinischer Sicht zu vermitteln. Ein fur den Routinebetrieb auf chirurgischen Stationen anwendbares Schema zur praoperativen Erfassung von Patienten mit erhohtem Blutungsrisiko sowie erste Erfahrungen damit werden vorgestellt.

Infectious disease marker testing is obligatory for the release of human tissue for transplantation. Most CE-marked tests are not validated for postmortem blood. In a previous study we have validated the testing for anti-HIV-1/2, anti-HCV, HBsAg, and anti-HBc. Here, we present the validation of testing for antibodies against T. pallidum, which is the last marker obligatory for tissue release for transplantation.17 samples of postmortem sera and 10 samples of both pre-und postmortem sera were obtained from cornea donors and tested for anti-T. pallidum on the Siemens-BEP-III-System. These sera were spiked with anti-T. pallidum-positive standard sera in concentrations which give low- and high-positive results at the respective dilution.Two of the unspiked postmortem sera were false-positive most likely due to intense hemolysis (free hemoglobin50 mg/dl). Of the 25 negative postmortem sera, none of the spiked samples was false-negative after 0, 24 and 60 h.There is no indication that postmortem samples give false-negative or false-positive results with the test system and test kits used in cases of low hemolysis. The procedure described might serve as a model for validating other test kits on postmortem samples.

BACKGROUND: Management of patients with clinically significant anti-IgA is difficult and unsatisfactory in many aspects. PATIENTS AND METHOD: A 40-year-old man with common variable immunodeficiency had a previous history of anaphylaxis after an intramuscular immuno- globulin administration. His serum contained anti-IgA, and he required immunoglobulins for recurrent infections. RESULTS: The administration of intravenous immuno- globulins (IVIgG) containing less than 0.1 mg per mL IgA led to an anaphylactic reaction after the transfusion of only 2 to 3 mL. The same IVIgG charge was subsequently pretreated with freshly separated autologous plasma and given to the patient on three consecutive days without any reaction (1.25, 10, and 10 g each in 400 mL plasma). Anti-IgA activity did not increase, and the patient was treated again without complications. DISCUSSION: Ex vivo pretreatment of IVIgG preparations with autologous plasma appears to be safe and useful in the management of patients with clinically significant anti-IgA. To achieve a significant IgA blockage, the preparation to be used should not contain large amounts of IgA. CONCLUSION: The strategy described here appears to be safe and may help prevent anaphylaxis in many instances.

Tissue and cell therapy is a field in which intense worldwide exchange is taking place. The European Community has therefore guaranteed to achieve the highest possible level of protection to safeguard public health regarding quality and safety of tissues and cells. The relevant Directive 2004/23/EC of the European Parliament and of The Council on Setting Standards of ‘Quality and Safety for the Donation, Procurement, Testing, Processing, Preservation, Storage and Distribution of Human Tissues and Cells’ and their complementary Directives 2006/17/EC and 2006/86/EC are now implemented into national law in the countries of the European Union. Alongside harmonized documentation of anamnesis and medical histories, testing for infection markers has always been of particular interest to physicians and regulators. Production and examination of tissue preparations have been adapted to the guidelines issued by authorities and specialist associations in the respective fields (musculoskeletal tissues, corneal tissue, heart valves), and evidence-based indication for transplantation has been discussed. In the present issue of Transfusion Medicine and Hemotherapy the reader is introduced to current facets of tissue banking seen from scientific points of view.