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H. W. Reesink, J. Lee, A. Keller, P. Dennington, J. Pink, R. Holdsworth, H. Schennach, M. Goldman, T. Petraszko, J. Sun, Y. Meng, K. Qian, V. Rehacek, P. Turek, T. Krusius, E. Juvonen, P. Tiberghien, D. Legrand, G. Semana, J. Y. Muller, J. Bux, A. Reil, C. K. Lin, H. Daly, E. McSweeney, L. Porretti, N. Greppi, P. Rebulla, H. Okazaki, S. A. Sanchez-Guerrero, H. A. Baptista-Gonzalez, C. Martinez-Murillo, A. Guerra-Marquez, H. Rodriguez-Moyado, R. A. Middelburg, J. C. Wiersum-Osselton, A. Brand, C. van Tilburg, D. Dinesh, J. Dagger, P. Dunn, E. Brojer, M. Letowska, K. Maslanka, E. Lachert, M. Uhrynowska, E. Zhiburt, M. Palfi, G. Berlin, B. M. Frey, L. Puig Rovira, E. Muniz-Diaz, E. Castro, C. Chapman, A. Green, E. Massey, N. Win, L. Williamson, C. C. Silliman, D. J. Chaffin, D. R. Ambruso, N. Blumberg, P. Tomasulo, K. J. Land, P. J. Norris, O. C. Illoh, R. J. Davey, R. J. Benjamin, A. F. Eder, L. McLaughlin, S. Kleinman & S. Panzer

Disuse atrophy of swallowing-related organs is suspected when decreased swallowing frequency is seen in the elderly. However, swallowing frequency has not been examined in elderly people during daily life. We developed a swallowing frequency meter containing a laryngeal microphone that does not restrict the subject's ability to perform daily activities. In this study, the utility of the meter was assessed. Experiment 1: The ability of the meter to detect swallowing was examined. The subject was instructed to swallow saliva or foods at a voluntarily pace. During these procedures, swallowing events were simultaneously recorded by the meter, self-enumeration and videofluorography. As a result, all of the swallowing events identified by the meter coincided with the swallowing events identified by self-enumeration and videofluorography. Experiment 2: Swallowing sounds display various patterns both between and within individuals. Therefore, we examined the concordance rate between the number of swallowing events counted by the meter and that counted by self-enumeration in 15 subjects over a longer period than in experiment 1. The concordance rates calculated by two examiners between the meter and self-enumeration were 96·8 ± 4·5% and 98·9 ± 3·3% at rest and 95·2 ± 4·5% and 96·1 ± 4·1% during meals, respectively. Our findings indicate that this meter is useful for measuring the frequency of swallowing during daily situations.

Transfusion-related acute lung injury (TRALI) is one of the most serious complications of blood transfusion. We have been gathering information about TRALl in Japan since 1997 through our national haemovigilance system. Recently, we have 20–40 TRALI cases per year (five million bags issued per year). Although TRALI is a rare complication of transfusion, it may become severe and may sometimes be fatal. A substantial number of TRALI cases have been caused by donor HLA or HNA antibodies, which were generated in alloimmunized, i.e. parous females. Considering this mechanism, TRALI mitigation strategies such as the use of male-only-plasma have been implemented in many blood centres or countries. Because of the availability of male plasma and the time limit for the production of fresh frozen plasma (FFP), we first tried to prepare FFP-LR2 derived from 400 ml whole blood only from male donors. FFP-LR2 accounts for 75% of FFP in Japan. (Six per cent of FFP products are from 200 ml whole blood and 19% from apheresis plasma.) A preliminary project on male-predominant plasma was started at several core blood centres. To minimize the burden on nurses regarding marking bags collected from males or females, which might lead to serious errors in collection sites, we updated the computer system that allows the staff members in the production department to sort male and female blood automatically. In a month after the system implementation nationwide, we successfully increased the production rate of male plasma for FFP-LR2 product by up to more than 98%. Regarding FFP-LR1 derived from 200 ml whole blood, the percentage of female donors is approximately 75%. One of the core blood centres has successfully implemented the same strategy for producing FFP-LR1, but others have not done so yet. Although male-only-plasma strategies achieved good results without any expensive screening tests to reduce TRALI incidence in some countries, the standard measures for other plasma rich blood products such as platelet concentrates or apheresis plasma have not been established yet. There are some measures to consider reducing the risk of TRALI caused by other plasma-rich products – recruiting only males as apheresis platelet donors, testing a certain number of donors for HLA antibodies, replacing the supernatant of a platelet pool with male plasma and excluding female donors from apheresis donations. Some blood centres adopted some of these measures, but so far there is little evidence for the efficacy of these measures for reducing the risk of TRALI. We have recently conducted research about the relationship between strength of HLA antibodies and TRALI development. On the basis of the results of the research, we can set the cut-off level in HLA antibody tests, which can reduce the risk of TRALI by eliminating strong HLA antibodies that presumably cause TRALI development if the specificity of an antibody matches the patient’s antigen. Preliminary screening for donor HLA antibodies is ongoing in three core blood centres in Japan, results of which enable us to establish effective and feasible screening strategies for HLA antibodies in the near future.

Transfusion-related acute lung injury (TRALI) is a life-threatening adverse reaction to transfusion. It is characterized by non-cardiogenic pulmonary edema that develops during or within 6 h after transfusion. Transfusion of blood products or massive transfusion has long been known as a rare cause of acute lung injury (ALI)/acute respiratory distress syndrome (ARDS). Because the main causes of ALI/ARDS are sepsis and/or pneumonia, little attention has been paid to transfusion as a cause of lung injury. Owing to indifference to and underrecognition of this complication, ALI after transfusion may be attributed to other causes. Moreover, physicians tend to attribute the cause of respiratory failure to more common complications of transfusion such as allergic reaction and circulatory overload. Even if an appropriate diagnosis of non-cardiogenic pulmonary edema is made, transfusion may not be identified as its cause. Although anti-leukocyte antibodies have been proved to be one of the main causes of TRALI, the precise mechanism of this syndrome is poorly understood. Anti-HLA and -HNA antibodies in labile blood products are reported to cause TRALI, and animal models of TRALI show that these antibodies induce lung injury under certain experimental conditions. We have recently showed that anti-CD36 antibody caused TRALI in two patients. This finding suggests that as-yet-unknown antibodies other than anti-HLA or -HNA antibodies might cause TRALI. According to current haemovigilance reports and previous look-back studies, the incidence of TRALI is not very high. This finding supports the concept of the two-hit theory of ALI, where certain patients’ clinical conditions are prerequisite in addition to transfusion for developing TRALI. A recent study in the ICU of a tertiary care medical centre has revealed a high incidence of suspected TRALI cases. This study revealed that female plasma, number of pregnancies in donors, number of donor units positive for anti-granulocyte antibodies and anti-HLA class II antibodies and concentration of lysophophatidylcholine in donor products are associated with developing TRALI. Our recent prospective study has revealed the predominance of male Fresh frozen plasma (FFP) FFP in post-transfusion respiratory function when comparing male only FFP and mixed FFP in surgical patients. The incidence of TRALI in the UK was successfully reduced by using preferentially male plasma. Although TRALI has a relatively better prognosis than ALI with other causes, TRALI has still been the leading cause of transfusion-related death in the United States in recent years. The prevention of TRALI caused by plasma products has been successful, but TRALI caused by apheresis platelets is still problematic. A recent study has suggested that the prevalence of anti-HLA antibodies in female donors increases with the number of pregnancies. Thus, screening for anti-HLA antibodies has emerged as an effective measure to prevent TRALI caused by apheresis platelets. At present, no single measure can thoroughly eliminate TRALI; however, early diagnosis and intervention may improve the outcome for patients with TRALI. Further study is required to elucidate the mechanisms of TRALI, which may lead to the development of preventive measures and treatment.

Recently, evidence implicating the roles of the anti-human leucocyte antigen (HLA) class II antibody in the development of transfusion-related acute lung injury (TRALI), which is one of the most serious possible side effects of transfusion, has been accumulating. The aim of this study is to clarify the roles of the anti-HLA DR alloantibody in TRALI development. Cultured human lung microvascular endothelial (LME) cells were incubated with either HLA-DR15-positive or HLA-DR15-negative monocytes together with serum from a single multiparous donor previously implicated in a clinical case of TRALI and known to contain anti-HLA DR15 antibody. Production of soluble leukotriene B(4) (LTB(4)) was measured in the supernatant and found to be markedly increased in the presence of HLA-DR15-positive monocytes but not with the HLA-DR15-negative monocytes or in the absence of LME cells. The vascular cell adhesion molecule-1 expression in LME cells and leucocyte-function-associated molecule-1 (LFA-1) expression in HLA-DR15-positive monocytes were notably enhanced after combined culture of LME cells, HLA-DR15-positive monocytes and TRALI-inducing anti-HLA DR15 antibody-positive serum. In conclusion, anti-HLA DR alloantibodies may be implicated in LME dysfunction that leads to TRALI, in a monocyte-dependent manner.

The structure of α-AgI which has a so-called average structure is studied by X-ray diffraction method. The obtained X-ray profile is composed of both, Debye peaks and diffuse haloes. The Debye peaks are explained qualitatively by “small divided cell model”. This model is also applied to the explanation of the absolute intensities of Debye peaks measured by Hoshino. The diffuse haloes are like those which are seen in the X-ray profile of a typical liquid metal so that they are analysed in a similar way as that of liquid metals. This analysis is consistent with the “small divided cell model”. The correlation between silver ions is relatively strong between the nearest neighbours, and the resultant coordination number of silver ions around a silver ion is 5.0. Die Struktur des α-AgJ, das eine sogenannte durchschnittliche Struktur hat, wird mit der Rontgenbeugungsmethode untersucht. Das Rontgen-Strahlenprofil von α-AgJ besteht sowohl aus Debye-Maxima als auch diffusen Haloes. Die Debye-Maxima werden vom Modell der „kleinen geteilten Zelle” qualitativ erklart. Dieses Modell wird auf die Erklarung der absoluten Intensitaten der Debye-Maxima, die von Hoshino gemessen wurden, angewendet. Die diffusen Haloes sind denen ahnlich, die im Profil eines typischen flussigen Metall analysiert werden. Diese Analyse steht nicht im Widerspruch zum Modell der „kleinen geteilten Zelle”. Die Korrelation zwischen Silberionen ist verhaltnismasig stark zwischen den nachsten Nachbarn, und die sich ergebende Koordinationszahl der Silberionen um ein Silberion ist funf.

The physicochemical properties of the purified calf thymus poly(ADP-ribose) polymerase were investigated. The enzyme purified to homogeneity was shown to contain about 10% DNA on a weight basis and its activity to be DNA independent. After removing this fragment of DNA, called the sDNA fraction, the enzyme becomes DNA dependent. The activity of this enzyme preparation was entirely dependent on, and completely restored by, added calf thymus DNA or sDNA. However, the calf thymus DNA concentration needed was a hundred times higher than that of sDNA. The properties of the two enzyme preparations, DNA independent and DNA dependent, were essentially the same. They both reacted against the specific antibody obtained with the DNA-independent poly(ADP-ribose) polymerase. The pH optimum was around 8; the activity was stimulated by Mg2+, Mn2+ and Ca2+, and inhibited by high ionic strength, p-chloromercuribenzoate, ADP-ribose, AMP and polylysine. Nicotinamide, thymidine and NADP were shown to be competitive inhibitors. The enzymatic activity was stimulated by histone H1 when the ratio of DNA to histone H1 was 2. Histones H2A, H2B, H3 and H4 had little effect on the DNA-independent enzyme activity, but were strongly inhibitory for the DNA-dependent enzyme. This inhibitory effect could be reversed by allowing the DNA-dependent enzyme to react with the sDNA fraction before adding histone subfractions. The apparent Km for NAD of the DNA-dependent poly(ADP-ribose) polymerase was shown to vary with the DNA concentration. It was minimum when the amount of sDNA was 10% of that of the enzyme. The ratio of the apparent Km for sDNA to the enzyme concentration was constant at any enzyme concentration. The minimum estimation of the number of base pairs of sDNA required for maximal activation of one enzyme molecule was 16. For calf thymus DNA, this estimation was of 640. These results suggest that the activation of the enzyme needs the formation of some complex between the protein and a specific part of the DNA. This complex was preserved in the DNA-independent enzyme preparation.

A processing protease has been purified from the matrix fraction of rat liver mitochondria. The purified protease contained two protein subunits of 55 kd (P-55) and 52 kd (P-52) as determined by SDS-PAGE. The processing protease was estimated to be 105 kd in gel filtration, indicating that the two protein subunits form a heterodimeric complex. At high ionic conditions, the two subunits dissociated. The purified processing protease cleaved several mitochondrial protein precursors destined to different mitochondrial compartments, including adrenodoxin, malate dehydrogenase, P-450(SCC) and P-450(11 beta), but the processing efficiencies were different each other. The endoprotease nature of the processing protease was confirmed with the purified enzyme using adrenodoxin precursor as the substrate; both the mature form and the extension peptide were detected after the processing. The processing activity of the protease was inhibited by metal chelators, and reactivated by Mn2+, indicating that the protease is a metalloprotease.