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Sera of 1046 patients undergoing haemodialysis for terminal renal failure or after renal transplantation were tested with the ELISA screening test for anti-LAV/HTLV-III and, if positive, the Western blot and ELAVIA tests. Four patients (0,38%) had "true" antibodies against LAV/HTLV-III, confirmed by the Western blot and ELAVIA tests. These patients had definite signs of cellular immune defects. They had received transplants from drug addict donors. Such kidneys should therefore no longer be used for transplantation. In addition, 29 patients (2.8%) had "false-positive" antibodies against LAV/HTLV-III in the ELISA test, unconfirmed in the Western blot and ELAVIA tests. The "false-positive" result was presumably due to cross-reaction with HLA antibodies. Sera of dialysis and transplantation patients who had received many blood transfusions should therefore be especially carefully tested before a diagnosis of infection with LAV/HTLV-III is made.

In a retrospective study the rate of HIV (LAV/HTLV-III)-infected recipients of blood units (excluding haemophiliacs) issued before HIV antibody testing became routine in 1985 was investigated. Screening of 200,000 donations yielded 24 seropositive blood donors. Of these 17 were homo- or bisexual, one subsequently known to take drugs by i.v. injection, one had received donor blood, and in five there was no information concerning sexual preference. Ten seropositive multiple blood donors (mean of 17.6 donations) and the 42 potentially infected units donated between 1982 and 1985 were examined in detail. Blood donations from infected first-time donors were discarded. Nine recipients were found to be seropositive (one case of transfusion-associated AIDS), five were seronegative. Both groups were evaluated clinically and immunologically. Ten recipients had died in the intervening time, and in 15 cases it proved impossible to determine whether the donated and incriminated blood had in fact been transfused. It is likely that there will be an increase in transfusion-associated AIDS over the next 3-5 years, before preventive measures of HIV antibody screening and voluntary exclusion of donors at risk will prove to be effective.

With the PALM MicroBeam system, precise laser microdissection of single cells from cell smears or tissue preparations is possible. Furthermore, this system uses a contact-free and therefore contamination-free laser pressure catapulting technique in which high energy generated by a focused laser pulse catapults single dissected cells into a collecting vessel. In this study, this technique was tested for forensic purposes with smear preparations from postcoital vaginal swabs, sperm swabs, and buccal cell swabs on different types of microscopic slides. Apart from super-frosted slides, cutting and catapulting of selected cells was possible in all cases. Subsequent polymerase chain reaction was performed using the genRES MPX-2 Amplification Kit. In the case of sperm cells stained with hematoxylin and eosin, fragments larger than approximately 200 bp could not be detected. Partial genetic profiles were obtained for DNA amounts originating from only two cell equivalents. Complete profiles, however, were observed with all preparations of a minimum of 10 epithelial cells, demonstrating a potential benefit of this technique for the contamination-free forensic analysis of extremely small specimens or mixed stains.

In this study, the effect of sample purification on total signal intensities of genRES® MPX-2 (nine-locus multiplex) amplified DNA prior to capillary electrophoretic analysis has been investigated. Sample purification with the QIAQUICK PCR purification kit led to an increase of the relative fluorescent signal intensity by a factor of 3.8±0.8. In addition, increase of injection time showed a linear increase of signal intensity between 3 and 10 s. Higher injection times may influence electrophoretic fragment separation and, therefore, lead to broad signals which may not be sized correctly.

D. J . Anstee, G. W. G. Bird E. Brodheim, J.-P. Cartron W. Dahr, C. P. Engelfriet P. D. Issitt, J . Jgrgensen L. Kornstad, M . Lewis C. Levene, A . Lubenko D. Mallory, P. Morel R . Nordhagen, Y. Okubo M . Reid, Ph. Rouger Ch. Salmon, S. Seidl, I? Sistonen S. Wendel, G. Woodfield 7: Zelinski International Society of Blood Transfusion Socihte lnternationale de Transfusion Sanguine Section Editor: B. Habibi, Paris

Aims: Leptin is a cytokine-type peptide hormone, recently implicated as a putative state marker of alcohol use and in craving. Our goal was to evaluate the potential of leptin as a state and trait marker and to rule out the role of current alcohol intoxication on leptin levels. Methods: Eighteen alcohol withdrawal patients (16 males, 2 females) whose blood contained 202 mg/dl (median) of ethanol at hospitalization, who had a median age of 43.5 years and had consumed 1075 g of ethanol (median) in the last 7 days were included in the study. Leptin was determined in samples at day 1 (when still intoxicated) and day 7 of withdrawal. Expected leptin levels were calculated with a formula. For comparison, 27 blood samples of 18 abstinent persons, matched for gender, age and body mass index were used. Furthermore, mean cell volume, γ-glutamyl transferase (GGT), blood glucose, cholesterol, triglycerides and body composition (bioimpedance device) were determined. For statistical analysis, SPSS 11 was used. Results: Expected leptin levels were 1.71 ng/ml (median), leptin measured at day 1 was 2.65 ng/ml (median) and 2.85 ng/ml on day 7 for the alcohol withdrawal patients and 2.2 ng/ml (median) for the abstainers. These concentrations were not significantly different. Significant correlations were found between leptin day 1 and expected leptin levels, percentage fat body mass, cigarettes smoked per day, GGT and blood alcohol concentration. Conclusions: Our preliminary data do not support the hypothesis of leptin as a state or trait marker and suggest only a minor influence of acute intoxication on leptin levels in alcohol detoxification patients.

During the occasional testing of Escherichia coli from faecal samples of young calves we observed multi-resistant isolates. Because of the significance of E. coli as an indicator bacterium for resistance trends we tested E. coli populations of young calves over a longer period. Here we present the results of a retrospective study comparing isolates from 1998 to 2000. Moreover, we compared, in a clinical study, the resistance rates of E. coli populations from 67 hospitalized calves both before and after hospitalization (with or without anti-microbial therapy), and with their anamnestic data of antibiotic usage. The highest resistance rates were found to be more than 80% for tetracyclines, ampicillin, sulfonamide/trimethoprim combinations, and chloramphenicol. A significant increase or decrease over the years was not observed. In analysing the data of hospitalized calves, an increase of resistance to some anti-microbials had to be registered that seemed to be connected with the selective pressure due to agents used in the clinic. In comparing anamnestic data and resistance rates it became obvious that reliable data are not easily available and that a number of potential anti-microbial influence factors have to be taken into account.

Ethyl glucuronide (EtG) is a non-volatile, water-soluble, stable-upon-storage, direct metabolite of ethanol and can be detected in body fluids and tissues (and also in post-mortem material) for an extended time period after the complete elimination of alcohol from the body (Alt et al. , 1997; Schmitt et al. , 1997; Seidl et al. , 1998; Wurst et al. , 1999 a , b ). The aim of the present Letter is to emphasize through our preliminary results that the detection of alcohol consumption that has taken place several weeks or months earlier, via the determination of EtG in hair, could be a useful forensic tool as well as for monitoring alcohol consumption of those in treatment for alcohol dependence. Thirty-one hair samples from four groups of subjects were analysed: (1) hair was taken post mortem from 16 subjects with a known history of alcoholism; the diagnosis was established by police investigations including …

HLA-DR specificities in 72 Addison's (AD) patients and 808 local controls were compared. We confirmed earlier reports that the HLA-DR3 specificity is significantly increased in AD patients. In our study a relative risk of 3.4 chi 2 = 22.5; pc = 0.01) for the disease was calculated. Analysis of HLA-DQB1 alleles in DR4+ Addison's patients with diabetes mellitus (N = 6) and without IDDM (14 of 18 individuals tested) revealed that the HLA-DQw8 allele (DQB1*0302) was significantly increased in AD patients with IDDM (chi 2 = 13.5; p = 0.001); conversely, a clustering of the HLA-DQw7 allele was detected in DR4+ Addison's patients without IDDM. We thus conclude that particular polymorphic alleles corresponding to non-charged amino acids at position 57 of the HLA-DQ beta-chain [non-Asp-57 alleles] are associated with IDDM also in Addison's patients.

Ethyl glucuronide (EtG) is a non-volatile, water-soluble, direct metabolite of ethanol that can be detected in body fluids and hair. We investigated urine and serum samples from three patient groups: (1) 33 in-patients in acute alcohol withdrawal; (2) 30 detoxified in-patients (treated for at least 4 weeks) from a 'motivation station'; and (3) 43 neuro-rehabilitation patients (non-alcoholics; most of them suffering from stroke, traumatic brain injury, Parkinson's disease etc.) using gas chromatography/mass spectrometry (GC/MS) with deuterium-labelled EtG as the internal standard and additionally in the second group of patients using liquid chromatography (LC/MS-MS). We found no correlation between the concentration of EtG in urine at hospitalization and the blood-ethanol concentration (r = 0.17), the time frame of detection (r = 0.5) or the total amount of clomethiazole required for the treatment of withdrawal symptoms (r = 0.28). In four out of 30 in-patients from the 'motivation station'--where neither clinical impression nor routine laboratory findings gave indications of relapse--concentrations of EtG in urine ranged between 4.2 and 196.6 mg/l. EtG concentrations in urine of between 2.89 and 23.49 mg/l were found in seven out of 43 neuro-rehabilitation patients using GC/MS. The GC/MS and the LC/MS-MS results showed a correlation of 0.98 with Pearson's correlation test and 1.0 with Spearman's correlation test. We suggest that EtG is a marker of alcohol consumption that can be detected for an extended time period after the complete elimination of alcohol from the body. When used as a relapse marker with a specific time frame of detection intermediate between short- and long-term markers, EtG fills a clinically as well as forensically important gap. Its specificity and sensitivity exceed those of all other known ethanol markers.

The prevalence of circulating autoantibodies against thyroperoxidase (mic-TPO) was determined in 3,000 healthy blood donors (age range: 23 to 60 years) from the Hamburg area. Of the blood donors, 153 (5.1 %) were found to have high titer of mic-TPO (> 350 IU/ml). Only two auto-antibody positive subjects (0.06%) were chemically hyper- and hypothyroid, respectively. Analysis of HLA-DR specificities revealed that HLA-DR specificities DR3 and DR5 were significantly increased when compared to controls (n=1,863). Comparison of the autoantibody-positive probands with a group of disease controls, i.e., Graves' patients and patients with lymphocytic thyroiditis, revealed a higher prevalence of HLA-DR3-positive HLA haplotypes in the disease controls when compared to autoantibody positives. Individuals with a mic-TPO level greater than 2,000 IU/ml were almost exclusively found to have one HLA-DR3 or HLA-DR5 positive HLA haplotype. We conclude that a high prevalence of hightiter mic-TPO can be found in healthy blood donors. Circulating signs of thyroid autoimmunity were associated with HLA specificities also found to be associated with autoimmune thyroid diseases.

Graves' disease (GD) is a human leukocyte antigen (HLA) linked organ-specific autoimmune disease. In German GD patients the disease is associated with HLA specificities of the HLA-DRw52 family (HLA-DR3, -DR5, and DR6; HLA-DRB3 positive HLA haplotypes). Recently, a strong association with a HLA-DRB3 restriction fragment length polymorphism gene has been described. To study HLA-DRB3 alleles and their association with the disease, a large cohort of controls (n = 3724) and GD patients (n = 304) was analyzed. HLA-DR allelic combinations revealed an increase in HLA-DR3/DR5 heterozygous patients (relative risk 2.9; P

A new polyvinyl chloride (PVC) blood storage container, PL 2209, plasticized with butyryl-n-trihexyl-citrate (BTHC) instead of diethylhexyl phthalate (DEHP) was used for in vitro and in vivo studies. Whole blood and red cell concentrates drawn on CPDA-1 were stored 42 days at 1-6 degrees C. Various biochemical parameters were tested and compared to those values seen in the literature for products stored in DEHP-plasticized containers. Measurements of lactate production, glucose consumption, sodium and potassium ions for both whole blood and red cells were compatible with published data. Adenosine triphosphate (ATP) values were on the average 73% for the red cell units and 80% for the whole blood units after 35 days storage. Hemolysis was 0.43% for the red cell concentrate and 0.38% for the whole blood units after the same storage period. Autologous recovery studies after 35 days of storage for both the whole blood units and red cell concentrate gave 80% recovery 24 h after reinjection. From these data it can be concluded that PL 2209 BTHC-plasticized PVC blood storage containers allow 35 days storage of blood draw on CPDA-1 and can be considered as an alternative to DEHP-plasticized PVC containers.

A Bombay blood group was detected in a 12-year-old Turkish girl with the rare Lewis type Le (a-b-). When using several phytohemagglutinins and Helix agglutinins (extracts from albumin glands of different snails), no ABH substances were detectable. However, after RDE treatment cryptantigens were demonstrable with anti-HLa (Laburnum alpinum), anti-BHEe (Evonymus europaeus), anti-AHp (Helix pomatia), and anti-TAh (Arachis hypogaea). Similarly, by application of the indirect fluorescence antibody method, only cryptantigens were detectable by anti-BHEe, anti-AHP, and anti-TAh. Therefore, it was assumed that in Bombay cells only the surface antigenic sites are absent, whereas the cryptantigens are present. Survival studies with autologous transfused 51Cr-labelled cells showed a normal lifespan (51Cr T/2 26d).

In einer Vergleichsuntersuchung wurden die Seren von 12080 unbezahlten Blutspendern mit dem Enzym-Immunoassay (EIA) und dem Radioimmunoassay (RIA) auf HBs-Antigen untersucht. Im EIA-Screeningtest wurde ein relativ hoher Prozentsatz nicht reproduzierbarer positiver Reaktionen festgestellt, der im wesentlichen durch ungenugendes Waschen bedingt ist. Wurde der EIA mit einer automatischen Waschapparatur durchgefuhrt, so konnte dieser Prozentsatz deutlich gesenkt werden. Nach Neutralisation wurden 0,31% positive Proben im EIA und im RIA festgestellt. Daraus kann geschlossen werden, das praktisch keine Unterschiede in der Empfindlichkeit zwischen diesen beiden Tests bestehen. Der EIA benotigt eine langere Inkubationszeit als der RIA. Der EIA hat jedoch den Vorteil, das er nicht mit radioaktiven Substanzen arbeitet, keine kostspielige Ausrustung erfordert und das die Resultate sofort ablesbar sind. Der EIA ist ohne Schwierigkeiten durchzufuhren, zwei med.-techn. Assistentinnen konnen etwa 1000 Seren pro Tag untersuchen.

Anti-ce (f) as a cause of haernolytic disease of the newborn was detected in the serum of a mother having the genotype CDe/cD-.

In a survey on 82,383 blood donor specimens from Hessen, West Germany, the observed percentage of repeatedly positive HTLV-III/LAV antibody ELISA results was 0.2%, but only 0.018% were confirmed by Western blot, immunofluorescence assay or radioimmunoprecipitation assay. These data are consistent with larger epidemiological data compiled from other German blood banks and suggest a high degree of nonspecific false-positive results; one of the causes for false-positives are HLA-DR4 antibodies, which react in the HTLV-III/LAV test. Epidemiological data on 127 dialysis patients and 1,153 prison inmates are presented.

Resume Il est generalement admis que pour la detection de l'antigene HBs, seules les techniques ayant une sensibilite de 3 e generation devraient etre employees. Celles-ci comprennent les tests radioimmunologiques (DRI) et l'hemagglutination passive inverse (RPHA). Recemment, deux techniques sensibles ont ete developpees : il s'agit des techniques enzymatiques (EIA) et d'agglutination de particules de latex. Ces deux techniques sont considerees comme appartenant aux tests de 3 e generation. Une etude comparative a ete entreprise afin de determiner quelle technique de 3 e generation etait la mieux adaptee au depistage chez les donneurs et laquelle etait la mieux adaptee aux Equipements Groupamatic. Les techniques RPHA sont legerement moins sensibles que les techniques DRI. En ce qui concerne la sensibilite, aucune difference n'a ete observee entre les techniques DRI et EIA. Cependant, aucune de ces methodes ne peut etre adaptee aux Equipements Groupamatic. Il etait donc particulierement interessant d'etudier dans le detail un test latex (Latex TG Pfizer). Plus de 3 000 serums provenant de donneurs volontaires ont ete testes simultanement en DRI et en Latex TG. Tous les positifs DRI ont ete detectes par le Latex TG. De plus, un panel constitue de 50 serums positifs en DRI (dont le taux d'antigene HBs etait variable d'un echantillon a l'autre), et de 20 serums negatifs, a egalement ete teste par ces deux techniques. Ici encore, aucune difference n'a ete observee a partir de trois lectures independantes. Cependant, avec un serum contenant un taux d'antigene HBs assez eleve, un phenomene de prozone fut observe ; il serait necessaire d'effectuer une etude approfondie de ce phenomene. En depit de ce fait, le reactif utilisant le latex peut etre, en ce qui concerne la detection de l'antigene HBs, la solution pour une technique tres sensible et adaptee aux Equipements Groupamatic.

In a comparative study 5,569 sera from voluntary donors were tested simultaneously for hepatitis B antigen by three reverse passive haemagglutination (RPHA) techniques (Auscell®, Hepanosticon®, Hepatest®) and the radioimmunoassay (RIA) (Ausria II). Among the three RPHA methods, no significant differences could be found as far as sensitivity is concerned. The percentage of positive sera detected by these RPHA methods varies from 0.43 to 0.56%. Eleven sera were detectable only by Ausria II. Except for two, all these sera had low counts per minute in the RIA.

Summary Several « third generationtests were compared: 5 reverse passive haemagglutination methods (RPHA), 2 enzymeimmunoassays (EIA), 1 radioimmunoassay (RIA) and 1 latex test. As far as the sensitivity is concerned RPHA techniques are less sensitive than the RIA. However, differences exist among the different RPHA methods. RPHA methods which utilize human red cells demonstrated a better sensitivity than those which utilize animal red cells. The RIA is widely accepted for blood donor screening. The test is well standardized, highly sensitive and specific. Also the EIA has been found to be highly sensitive but showed a greater degree of non-specificity. Rapid HBsAg screening in blood donors can be performed by RPHA methods or by a latex test. For the latter Antigex TG was used. This test has a sensitivity which is similar to that of some of the RPHA methods. The most striking results was the observation that Antigex TG can cause a prozone phenomenon. This phenomenon can be avoided when the serum specimen was spread out before the latex reagent was added.

In the sera of 68 (20.9%) of 325 haemodialysis patients, a specific cold agglutinin, cross-reacting with anti-N, was detected. Each patient showing this anti-N-like antibody used a formaldehyde-sterilized dialyser. By contrast, the antibody was not found in any of the 73 patients using a non-formaldehyde-sterilized dialyser. The results of transfusion experiments indicate an in vivo haemolytic activity of the antibody. In vitro, formaldehyde pretreatment of erythrocytes increased the agglutinability of the cells by the anti-N-like antibody. This effect could be prevented when the MN receptor of the red cells was exposed to neuraminidase. Trace quantities of formaldehyde inevitably enter the patient during dialysis when a formaldehyde-sterilized dialyser is used. It is postulated that this in vivo formaldehyde exposure might interact with the MN receptor of the red cells, rendering it immunogenic, and thus induce the formation of the haemodialysis-associated anti-N-like antibody.

Two siblings with the rare phenotype Rhnull are described; the parents are related to each other. Serologic investigation of the family gives indirect support for the action of an amorphous gene. As demonstrated in the propositus, the splenectomy resulted in health improvement. Titration tests in 1,803 unrelated blood donors revealed 4 heterozygous — types. On the basis of this calculation one can assume one Rhnull patient among 6 million inhabitants.

Es wird uber einen Anti-Vel-Antikorper berichtet, der bei einer Patientin im Rahmen der serologischen Vertraglichkeitsprobe entdeckt wurde. Die Familienuntersuchung ergab, das eine Schwester der Patientin ebenfalls Velnegativ war. Auch bei dieser Schwester wurde eine Anti-Vel-Antikorper im Serum gefunden. In beiden Fallen waren Transfusionen und Schwangerschaften vorausgegangen. Die Antikorper waren hamolysierend, allerdings konnte die Hamolysinwirkung des schwacheren Serums nur mit enzymbehandelten Erythrozyten nachgewiesen werden. Etwa 10% der untersuchten Blutproben reagierten im NaCl-Milieu schwach positiv, bei Neugeborenen war dieser Prozentsatz um etwa das Doppelte erhoht. Bei der Untersuchung von 5371 unausgelesenen Blutproben fanden wir zwei Vel-negative.