Karin Janetzko

Therapeutic efficacy and safety of photochemically treated apheresis platelets processed with an optimized integrated set.

BACKGROUND: This multicenter, randomized, controlled, double‐blind Phase III clinical study evaluated the therapeutic efficacy and safety of apheresis platelets (PLTs) photochemically treated (PCT) with amotosalen and ultraviolet A light (INTERCEPT Blood System, Baxter Healthcare Corp.) compared with conventional apheresis PLTs (reference).

Fully automated processing of buffy-coat–derived pooled platelet concentrates

BACKGROUND:  The OrbiSac device, which was develo‐ped to automate the manufacture of buffy‐coat PLT con‐centrates (BC‐PCs), was evaluated.

Polymerase chain reaction inhibition assay documenting the amotosalen‐based photochemical pathogen inactivation process of platelet concentrates

BACKGROUND: The INTERCEPT Blood System (Baxter Healthcare Corp.) for platelets (PLTs) uses amotosalen‐HCl (S‐59) in conjunction with ultraviolet A (UVA) light to inactivate contaminating pathogens by modifying the nucleic acids of pathogens. The success of this photochemical treatment (PCT) process can be documented indirectly with a high‐performance liquid chromatography assay measuring the photodegradation of amotosalen and measurement of the UVA light dose delivered by the illumination system.

Effects of Blood Donation on the Physical Fitness and Hemorheology of Healthy Elderly Donors

Background: International regulations for blood donation recommend a maximum donor age of 65 years. As the average population age is steadily rising in western societies, a considerable group of volunteers is lost to the donor base. Study Design and Methods: In a prospective study we investigated the effect of a 450-ml whole blood donation on the physical fitness and hemorheology of regular elderly allogeneic blood donors (n = 24, aged 63–69 years, mean = 65). Results were compared with a younger group of regular donors (n = 23, aged 55–62 years, mean = 58) and a group of elderly subjects (n = 7, aged 63–66 years, mean = 65), who did not donate blood for this study. Assessing the physical fitness, we determined the submaximal physical working capacity at a heart rate of 130 min–1 (PWC 130) and the maximal working capacity (MWC) by treadmill exercise testing the day before (day –1) and after donation (day +1). The impact of the blood loss on hemorheology was examined by analyzing the plasma viscosity before, during and after donation. Results: We found an increase of mean values of PWC 130 and MWC on day +1 in all study groups, but increases were only significant in the younger group (PWC 130 p = 0.03; MWC p = 0.04). Values did not differ significantly between the three groups. Plasma viscosity decreased significantly directly after donation in both groups of donors. Conclusion: A single blood donation did not alter the physical fitness of otherwise healthy elderly people. The older blood donors and the younger controls showed a similar compensation mechanism to blood loss. We found no general reason for disqualifying blood donors aged 65 years from donating.

Coxiella burnetii - Pathogenic Agent of Q (Query) Fever

1.1.1 Structure C. burnetii is a member of the family of the Coxiellaceae bacteria and replicates intracellularly in cells of different species. Phylogenetically related bacteria include Legionellaceae, Francisellaceae, Pseudomonaceae, and other Gammaproteobacteria. Coxiella are small Gram-negative, pleomorphic, coccoid bacteria with a size of 0.2–1.0 m. They occur in 3 different forms: small cells (small cell variant, SCV) which are highly infectious, large cells (large cell variant, LCV) which develop also in cell culture, as well as spore-like particles (SLP) which are infectious and very robust to environmental conditions. Dependent on the host system, Coxiella undergoes a phase variation during growth [12]. In mammalian cells, bacteria grow as LCV, and form spore-like particles and 2 different antigenic forms described as Phase I and II.

Generation of neutrophil priming activity by cell-containing blood components treated with pathogen reduction technology and stored in platelet additive solutions

BACKGROUND: Storage of cell‐containing blood components such as platelet concentrates (PCs) and red blood cells (RBCs) results in generation of biologically active compounds, many of which may be associated with adverse transfusion events. Priming of the neutrophil oxidase activity is a common characteristic of many of the biologically active compounds found in stored blood. We evaluated the priming activity of pathogen reduction technology (PRT)‐treated PCs stored in plasma or platelet additive solution (PAS) and PRT‐treated RBCs.

A single-tube real-time PCR assay for Mycoplasma detection as a routine quality control of cell therapeutics.

Background: Contamination of cell culture and biological material by mollicute species is an important safety issue and requires testing. We have developed a singletube real-time polymerase chain reaction (PCR) assay for rapid detection of Mollicutes species stipulated by the European Pharmacopeia. Methods: Primers and TaqMan probes (FAM-labeled) were deduced from 16S rDNA sequence alignment of 18 mollicutes species. A synthetic internal control (IC) DNA and an IC-specific TaqMan probe (VIC-labeled) were included. The analytical sensitivity of the assay was determined on DNA dilutions from 12 mollicute strains. Specificity was proven by the use of DNA from other bacteria. Results: Analytical sensitivities of the PCR assay were in the range of 405-2,431 genomes/ml for 11 of the 12 tested mollicute DNA samples. The lowest sensitivity was found for Ureaplasma urealyticum (19,239 genomes/ml). Negative results for DNA samples from 3 different ubiquitous bacteria demonstrated the specificity of the PCR assay for Mollicutes. Direct testing of cell culture supernatants spiked with Mycoplasma orale revealed similar sensitivity compared to isolated DNA. Conclusions: Our single-tube real-time PCR assay with internal reaction control enables rapid and specific detection of mollicute contaminants. The test protocol is suitable for routine quality control of cell therapeutics.

Pathogen Reduction in Blood Products: What’s Behind These Techniques?

The collection, separation and transfusion of red blood cells, platelets, whole plasma and fractionated plasma components are mainstays of our health care system. Each of these elements is essential for the preservation of life and the treatment of disease. With the rise in blood transfusion in the middle of the last century it became clear to the medical community that these life-sustaining and essential therapeutic substances were transmitting life-threatening diseases. Beside immunological risks like febrile and nonfebrile reactions, acute and prolonged hemolytic reactions, transfusion-related acute lung injury (TRALI) or graft-versus-host disease (GvHD) suspicious of transfusion-associated infection raise up first in 1943 when transfusion-associated hepatitis was observed [1]. The introduction of a routine test took about 20 years so that the screening for HBV antigen could be integrated obligatory into the blood donation testing program in 1971. Up to now the implementation of different screening tests, especially NAT testing, for syphilis, HBV, HCV and HIV as well as partly HAV and parvo virus B19minimized the risk of viral transfusion-associated infection considerable. Thus currently the estimated risk for transfusion-associated HBV infection is 1:360,000, for transfusion-associated HCV 1:10.9 million and for transfusion-associated HIV 1:4.3 million [2]. Though the viral problem seems to be managed, one has to take into account that there is still a residual viral transfusion-associated risk due to pathogens for which actually no detection systems exist, e.g. arbovirus, the agent for chikun-gunya. Moreover, as we learned from the past, viruses still can switch their host in regular periods and move from animal to human pathogenicity such as Ebola (1977), HIV (1981), SARS (2003) or H5N1 virus (2006) [3, 4, 5]. It can be assumed that such pathogens can cause fatal diseases due to the transmission via blood. The challenging problem lies in the delayed development of suitable detection systems so that potentially contaminating agents passed undetected into the blood supply. The resulting concerns raised by these events triggered the development of techniques to reduce or even eliminate infectious agents from blood products. While in the middle of the 1990s the transmission of virus with blood products could be stemmed, the introduction of hemovigilance systems in different countries focused on bacterial transfusion-associated infections, a problem that attracted less attention in the years before. Meanwhile this has become the most frequent infection risk in transfusion medicine and is considered to be associated with a high rate of death from transfusion. Due to the storage condition the highest risk results for platelet products, and a prevalence for bacterial contamination of 1:1,500 per platelet unit was reported [6]. A frequency of 1 in 50,000 platelet transfusions was described for significant clinical events. The mortality rates for platelet-related sepsis range from 1:230,000 to 1:625,000 donor exposures [7, 8]. Based on these findings two major strategies are followed: i) the development of fast screening tests and ii) the development of techniques for inactivation or reduction of these pathogens. Screening tests are based on NAT targeting the uniform bacterial region 16S DNA [9], on FACS technique procedure detecting thiazol orange-stained bacteria [10] or on culturing methods [11]. Pathogen inactivation or reduction techniques came up first in the 1960s, and the methylene blue/light and the solvent-detergent treatment for plasma or plasma components were established. These techniques were not applicable for cellular blood products because cells are sensitive to such chemical or physical treatment. However, at the beginning of the 1990s new promising techniques were developed that use the circumstance that contaminating agents like bacteria, viruses, protozoa and also leukocytes need their DNA or RNA genome for cell function while red blood cell and platelet function is independent from a genome. This issue of Transfusion Medicine and Hemotherapy will give an overview on all pathogen reduction techniques which are still used in admitted blood therapeutics or which are actually on their way of development. The review articles written by expert authors summarize comprehensive information on the treatment using riboflavin and UV light [12], amotosalen and UVA light [13], S-303 [14], UVC light [15], methylene blue/light [16] and the solvent-detergent technique [17]. Each article is focused on the mechanism of action, toxicity, side effects, clinical efficiency of the treated blood component and the inactivation efficacy on different kinds of pathogens.

Development of Anti-G, Anti-C and Anti-Jk(b) in a 22-Year-Old Mother during Her Fourth Pregnancy

Background: Anti-G antibodies are rarely found since anti-D, in combination with anti-C, are difficult to discriminate from anti-G antibodies in routine testing. Case Report: A 22-year-old, gravida-3, para-1, woman with blood group A Rh D neg ccddee and known antibody anti-Jk(b), gave birth to her second child. While anti-Jk(b) could not be detected at birth, a new anti-C was found. Antibody screening tests (IAT) were performed using gel cards and rare G positive rGr erythrocytes. Genotyping for RHD and RHCE was performed using PCR-SSP. Results: The childs blood group was A Rh D neg Ccddee. Genotyping revealed Cde/cde haplotypes. The erythrocytes of the new-born showed a positive direct antiglobulin test with IgG; anti-D and anti-C could be eluted. Erythrocytes with the rare phenotype rGr were reactive with the serum of the mother. Conclusion: The presence of anti-D and anti-C in the eluate from then new-borns Ccddee erythrocytes proved anti-G or anti-G in combination with anti-D. When anti-C and anti-D are seen during a pregnancy, possibly anti-G is present. This observation is of relevance since women with anti-G can still develop anti-D and require rhesus prophylaxis.

Blutverlust bei älteren Probanden und die Folgen für Rheologie und kardiale Leistungsfähigkeit

Ziel: Die Auswirkung einer einmaligen hypovolamischen Blutspende auf die Rheologie und die ergometrisch bestimm-bare Leistungsfahigkeit bei klinisch gesunden alteren Blut-spendern w