Massive pulmonary hemorrhage before living donor liver transplantation in infants

A massive pulmonary hemorrhage in patients with liver cirrhosis is a life‐threatening complication that may result in a contraindication of a liver transplantation because of its high mortality rate. Herein, we present two infant biliary atresia cases that successfully underwent an LDLT that was followed by intensive respiratory care for the pretransplant massive pulmonary hemorrhage. Both cases exhibited severe respiratory failure (minimum PaO₂/FiO₂; 46 mmHg and 39 mmHg, respectively). To arrest the bleeding, we applied a very high positive pressure ventilation treatment (maximum PIP/PEEP; 38/14 cmH₂O and 55/15 cmH₂O, respectively), plasma exchange, several FFP transfusions, and recombinant factor VIIa via intrapulmonary administration. In addition, we used CHDF treatment, applied HFOV transiently, and treated the patient with inhalation of nitric oxide. Although we prepared ECMO for intra‐operative use, both cases were successfully managed with conventional mechanical ventilation without using ECMO, which may have worsened the pulmonary hemorrhage due to the use of an anticoagulant. Use of an excessive positive pressure management, although it poses a risk for barotrauma, could be acceptable to arrest the pulmonary bleeding in selected cases of liver failure patients who have no time remaining before LDLT.     

Regulation of FcepsilonRI-mediated degranulation by an adaptor protein 3BP2 in rat basophilic leukemia RBL-2H3 cells

Aggregation of high-affinity IgE receptor FcepsilonRI induces sequential activation of nonreceptor-type protein-tyrosine kinases and subsequent tyrosine phosphorylation of cellular proteins, leading to degranulation in mast cells. A hematopoietic cell-specific adaptor protein, 3BP2, that was originally identified as an Abl SH3-binding protein was rapidly tyrosine phosphorylated by the aggregation of FcepsilonRI on rat basophilic leukemia RBL-2H3 cells. Tyrosine phosphorylation of 3BP2 did not depend on calcium influx from external sources. To examine the role of 3BP2 in mast cells, we overexpressed the SH2 domain of 3BP2 in the RBL-2H3 cells. Overexpression of 3BP2-SH2 domain resulted in a suppression of antigen-induced degranulation as assessed by beta-hexosaminidase release. Even though overall tyrosine phosphorylation of cellular protein was not altered, antigen-mediated tyrosine phosphorylation of phospholipase C-gamma (PLC-gamma) and calcium mobilization were significantly suppressed in the cells overexpressing the 3BP2-SH2 domain. Furthermore, antigen stimulation induced the association of 3BP2-SH2 domain with LAT and other signaling molecule complexes in the RBL-2H3 cells. FcepsilonRI-mediated phosphorylation of JNK and ERK was not affected by the overexpression of 3BP2-SH2 domain. These data indicate that 3BP2 functions to positively regulate the FcepsilonRI-mediated tyrosine phosphorylation of PLC-gamma and thereby the signals leading to degranulation.     

Association of Gln222Arg polymorphism in the deoxyribonuclease I (DNase I) gene with myocardial infarction in Japanese patients.

AIMS: We have recently reported that serum deoxyribonuclease I (DNase I) activity, which may be involved in apoptosis, increases abruptly in the early phase of acute myocardial infarction (MI) [Kawai Y, Yoshida M, Arakawa K, Kumamoto T, Morikawa N, Masamura K, Tada H, Ito S, Hoshizaki H, Oshima S, Taniguchi K, Terasawa H, Miyamori I, Kishi K, Yasuda T. Diagnostic use of serum deoxyribonuclease I activity as a novel early-phase marker in acute myocardial infarction. Circulation 2004;109:2398-2400]. Death of vascular smooth muscle cells, in part because of apoptosis, is postulated to heighten susceptibility to disruption of vulnerable plaque, resulting in onset of MI. The present study evaluated the possibility that Gln222Arg polymorphism of the DNase I gene may be one of the factors involved in predisposition to MI. METHODS AND RESULTS: We assessed 611 Japanese patients: 311 with MI and 300 with stable angina pectoris (AP). Three common phenotypes determined by two common codominant alleles, DNASE1*1 and *2, whose corresponding gene products exhibit different properties, were found in these patient groups. The prevalence of DNASE1*2 was significantly higher in patients with MI than in those with AP (0.543 vs. 0.428, P < 0.001), being confirmed by phenotyping of the second study population. Multiple logistic regression analysis showed that the odds ratio of DNASE1*2 was 1.51 [95% confidence interval (CI) 1.04-2.18]. The association of the DNASE1*2 allele with MI was statistically significant, being independent of other conventional risk factors. CONCLUSION: Our data demonstrate that Gln222Arg polymorphism in the DNase I gene is associated with MI in the Japanese patients.     

Matrix metalloproteinase 9 (MMP-9) in patients with rheumatoid arthritis

Matrix metalloproteinase 9 (MMP-9) degrades type IV collagen, gelatin, type V collagen and type XI collagen. We measured proMMP-9 and proMMP-9-TIMP-1 complex in sera and joint fluids by sandwich ELISA, and immunohistochemically examined the expression of this enzyme in joint tissues from patients with rheumatoid arthritis (RA). ProMMP-9 was purified from the culture medium of HT 1080 cells by the three steps of chromatography. Purified proMMP-9 and activated MMP-9 by aminophenylmercuric acetate showed two bands of 92 and 67 kDa on gelatin zymography. We raised two monoclonal antibody clones, named 2G9 and 8G7, against proMMP-9. 2G9 and 8G7 reacted with proMMP-9 in western blotting and these clones reacted not only with proMMP-9, but also with proMMP-9-TIMP-1 complex in sandwich ELISA, respectively. The proMMP-9 concentration in 86 sera (749.4±940.2 ng/ml) and 54 joint fluids (4539.9±7681.5 ng/ml) from patients with RA was significantly higher than those of patients with osteoarthritis (15 sera: 139.0±149.6 ng/ml; 16 joint fluids: 655.0±1982.8 ng/ml) and control (37 sera: 266.7±120.4 ng/ml; three joint fluids: 0 ng/ml). The immunohistochemistry with 2G9 monoclonal antibody showed that proMMP-9 were expressed in the neutrophils and the monocytes-macrophages which diffusely infiltrated in the sublining layer of rheumatoid synovium. In addition, the osteoclasts along subchondral bone were also intensively stained. The proMMP-9 concentration in joint fluids from 39 RA patients was positively correlated to the count of proMMP-9 positive cells in RA synovium (r=0.607) and to the score of diffuse infiltrates of lymphocytes (r=0.720). However, it did not show correlation to the stage and the class defined by Steinbrocker and to the other clinical laboratory data. Our results suggest that proMMP-9 actively participates in joint destruction of RA through the expression of neutrophils and monocytes-macrophages and is regulated by lymphocytes.