Strong correlation between the prevalence of cerebral infarction and the presence of anti-cardiolipin/beta2-glycoprotein I and anti-phosphatidylserine/prothrombin antibodies--Co-existence of these antibodies enhances ADP-induced platelet activation in vitro

Cerebral infarction is the most common arterial thromboembolic complication in the anti-phospholipid antibodies (aPL) syndrome. In an effort to clarify the roles of aPL in the pathogenesis of cerebral infarction in patients with SLE, we examined the levels of anti-cardiolipin/2-glycoprotein I antibodies (anti-CL/beta2-GPI) and anti-phosphatidylserine/prothrombin anti-bodies (anti-PS/PT) in addition to lupus anticoagulant (LA) activity in 126 patients with SLE (35 with cerebral infarction and 91 without thrombosis). Both anti-CL/beta2-GPI and anti-PS/PT strongly correlated with the presence of LA activity. The prevalence of cerebral infarction was obviously higher in the patients who had both anti-CL/beta2-GPI and anti-PS/PT (76.5% [26/34 cases], p<0.0001) than in the other patients having anti-CL/beta2-GPI or anti-PS/PT alone or neither of them (9.8% [9/92 cases]). Furthermore, we studied the in vitro effects of anti-CL/beta2-GPI and/or anti-PS/PT on the enhancement of platelet activation induced by stimulation with a low concentration of adenosine diphosphate (ADP).The purified IgG containing both anti-CL/beta2-GPI and anti-PS/PT caused significant enhancement of platelet activation caused by ADP. However, the purified IgG containing either anti-CL/beta2-GPI or anti-PS/PT had no enhancing effects on it. Furthermore, platelet activation was generated by the mixture of anti-CL/beta2-GPI-IgG and anti-PS/PT-IgG prepared from individual patients, but not by each fraction alone. These results indicate that anti-CL/beta2-GPI and anti-PS/PT may cooperate to promote platelet activation, which may contribute to the risk of cerebral infarction in patients with SLE.     

Enhanced binding affinity for FcgammaRIIIa of fucose-negative antibody is sufficient to induce maximal antibody-dependent cellular cytotoxicity

Antibody-dependent cellular cytotoxicity (ADCC) is considered to be an important therapeutic function for clinical efficacy of monoclonal antibodies. Recent studies have revealed two methods to increase binding affinity for FcgammaRIIIa and enhance ADCC more efficiently for antibodies: (i) fucose removal from antibody N-linked complex oligosaccharides and (ii) amino acid mutations in the antibody Fc region. In this study, we compare the biological activities of the methods of generating high ADCC antibodies. We used a fucose-negative antibody and two antibodies with sets of mutations, demonstrated previously to optimally enhance ADCC using the chimeric anti-CD20 antibody, rituximab, as the model. Both amino acid mutant antibodies showed a significantly higher affinity for recombinant FcgammaRIIIa than fucose-negative antibody when compared using biosensor analysis. The removal of fucose from the antibodies bearing amino acid mutations exhibited a further enhancement of binding to recombinant FcgammaRIIIa and significantly increased binding to natural killer (NK) cells. Despite the differences manifested in binding for the FcgammaR, ADCCs were indistinguishable between methods and even when the methods were combined. These results indicate that the affinity of binding to FcgammaRIIIa does not predict ADCC beyond a certain threshold and that each method alone is sufficient to induce maximal ADCC of the antibody.     

Multi-level,cross-sectional study of workplace social capital and smoking among Japanese employees

Background  

Social capital is hypothesized to be relevant to health promotion, and the association between community social capital and cigarette smoking has been examined. Individual-level social capital has been found to be associated with smoking cessation, but evidence remains sparse on the contextual effect of social capital and smoking. Further, evidence remains sparse on the association between smoking and social capital in the workplace, where people are spending an increasing portion of their daily lives. We examined the association between workplace social capital and smoking status among Japanese private sector employees.     

Estimating medical expenditures spent on rule-out diagnoses in Japan

Background According to the regulations concerning reimbursement rules for the uniform coverage scheme in Japan's health insurance system, rule‐out diagnoses must be included in a health insurance claim (HIC) to ensure reimbursement for clinical procedures whose results show that a suspected disease is not present. However, estimations of disease‐specific medical expenditure by conventional methods have not considered the information on rule‐out diagnoses. Objectives To estimate disease‐specific medical expenditure for rule‐out diagnoses. Methods Data were obtained from 169 622 outpatient HICs in May 2006 from corporate health insurance societies. We used the proportional distribution method to estimate medical expenditure for each of the major disease categories defined by the Classification of Diseases for the use of Social Insurance, which is based on the International Statistical Classification of Diseases and Related Health Problems, 10th Revision. Results There were 442 010 diagnoses on the HICs, of which 20 330 (4.60%) were rule‐out diagnoses. Rule‐out diagnoses accounted for 8.5% of total medical expenditure. The proportion of medical expenditure spent on rule‐out diagnoses varied across the major diseases categories, and it was estimated that more than one‐third (36.9%) of the medical expenditure on neoplasm is spent on rule‐out diagnoses. Conclusions The existence of rule‐out diagnoses affects the estimation of disease‐specific medical expenditure. Therefore, the estimation of disease‐specific medical expenditure and evaluation of prevention and treatment programmes should be improved by utilizing information on rule‐out diagnoses.     

Outbreak of Salmonella Braenderup infection originating in boxed lunches in Japan in 2008

There have been only 2 reports of a large-scale foodborne outbreak arising from Salmonella enterica serotype Braenderup infection worldwide. On August 9, 2008, an outbreak originating in boxed lunches occurred in Okayama, Japan. We conducted a cohort study of 786 people who received boxed lunches from a particular catering company and collected 644 questionnaires (response rate:82%). Cases were defined as those presenting with diarrhea (≧4 times in 24h) or fever (≧38℃) between 12 am on August 8 and 12 am on August 14. We identified 176 cases (women/men:39/137);younger children (aged＜10 years) appeared to more frequently suffer severe symptoms. Three food items were significantly associated with higher risk of illness;tamagotoji (soft egg with mixed vegetables and meat) (relative risk (RR):11.74, 95% confidence interval (CI):2.98-46.24), pork cooked in soy sauce (RR:3.17, 95% CI:1.24-8.10), and vinegared food (RR:4.13, 95% CI:1.60-10.63). Among them, only the RR of tamagotoji was higher when we employed a stricter case definition. Salmonella Braenderup was isolated from 5 of 9 sampled cases and 6 food handlers. It is likely that unpasteurized liquid eggs contaminated by Salmonella Braenderup and used in tamagotoji caused this outbreak.     

Optimizing Therapeutic Antibody Function

Since the establishment of monoclonal antibody production using hybridoma technology in the mid-1970s, there has been expanding progress and continuous technological improvement in the development of therapeutic antibodies. The initial technological breakthroughs involved reduction of immunogenicity and thus enabled repeated administration. The establishment of chimeric, humanized, and fully human antibodies has led to the great success of several ‘second-generation’ therapeutic antibodies, such as rituximab, trastuzumab, cetuximab, and bevacizumab. However, there still exists an urgent demand for improvement in the efficacy of the current antibody therapeutics, which is not yet fully satisfactory for patients. Based on the current understanding of the clinical mechanisms of several therapeutic antibodies, many now believe that Fc-mediated functions (e.g. antibody-dependent cellular cytotoxicity, complement-dependent cytotoxicity, and neonatal Fc receptor [FcRn]-mediated storage) will improve the clinical outcomes of therapeutic antibodies. The present review focuses on the recent progress in the development of ‘Fc engineering,’ which dramatically improves (and sometimes silences) Fc-mediated functions. These achievements can be classified into two technological approaches: (i) introducing amino acid mutations and (ii) modifying Fc-linked oligosaccharide structures. The effectiveness of multiple third-generation therapeutic antibodies armed with various engineered Fcs is now ready to be tested in clinical trials.     

Effects of anticoagulants and storage temperature on immature platelet fraction % (IPF%) values in stored samples measured by the automated hematology analyzer, XE-5000--utility of CTAD-anticoagulation and room temperature storage

Measurement of reticulated platelet percentage (RP%) is thought to be a useful marker for differential diagnosis and analysis of platelet kinetics in patients with thrombocytopenic disorders. Two methods are used to detect RP; flow cytometric method and immature platelet fraction (IPF) method using automated hematology analyzers. Although IPF% measured by the automated hematology analyzers is simple and convenient, we already reported that IPF% values were highly fluctuated in stored whole blood sample with EDTA-2K at 4 degrees C day by day. In this study we investigated the stability of IPF% in blood samples obtained from 11 patients with chronic immune thrombocytopenic purpura (ITP) and 19 healthy volunteers using the automated hematology analyzer, XE-5000 (Sysmex) under various storage conditions. EDTA-2K, 3.13% sodium citrate, acid-citrate dextrose solution (ACD), citrate-theophylline-adenosine-dipyridamole solution (CTAD), or sodium fluoride was used as an anticoagulant. When blood samples obtained from healthy subjects were stored at 4 degrees C, IPF% values markedly increased in a time-dependent manner by any anticoagulant examined. On the other hand, there was no significant or only slight difference in IPF% values at room temperature (RT) storage except sodium fluoride. However, in patients with ITP the elevated IPF% values fluctuated widely in EDTA-2K, sodium citrate and ACD-anticoagulated samples even at RT storage. In contrast, IPF% values in CTAD samples stored at RT were highly stable in all patients with ITP up to 4 day storage. These results suggest that the measurement of IPF% by XE-5000 provides quite stable data up to 4 day-storage in ITP patients as well as healthy subjects under CTAD-anticoagulation and RT storage conditions.     

Prevalence of genetic mutations in protein S, protein C and antithrombin genes in Japanese patients with deep vein thrombosis

Introduction

Genetic deficiencies of PROS1, PROC, and SERPINC1 (antithrombin) are risk factors for deep vein thrombosis (DVT). Diagnosis of the inherited deficiencies of these three genes is sometimes difficult because of the phenotypic variability. This study was undertaken to reveal the frequency of nonsynonymous mutations of these three genes in Japanese DVT patients.

Patients/Methods

One hundred seventy-three DVT patients were registered by the Sub-group of Blood Coagulation Abnormality, from the Study Group of Research on Measures for Intractable Diseases. We sequenced the entire coding regions of the three genes in all DNA samples and identified the nonsynonymous mutations.

Results and Conclusions

For PROS1 we identified 15 nonsynonymous mutations in 28 DVT patients; for PROC, 10 nonsynonymous mutations in 17 patients; and for SERPINC1, 13 nonsynonymous mutations in 14 patients. Five patients had two mutations in PROS1 and PROC, and all of them had PROS1 K196E mutation. We previously identified one patient with a large PROS1 gene deletion. Thus, 55 out of 173 patients (32%) carried at least one genetic defect in the three genes. The PROS1 K196E mutation found in 15 Japanese DVT patients was the most prevalent. Mutations of PROC K193del and V339M were the second, each found in four patients. Our data suggested that the PROC K193del mutation caused the loss of the anticoagulant activity but not the amidolytic activity. Our effort is the first DNA resequencing study to identify the genetic variations in DVT patients without any consideration of their plasma activities and antigens. To minimize selection bias in a future evaluation of the contribution of genetic deficiency to DVT, we must recruit patients consecutively.     

Contrast-enhanced ultrasonography depicts small tumor vessels for the evaluation of pancreatic tumors

OBJECTIVE: The aim of this study is to evaluate the efficacy of contrast-enhanced ultrasonography for the diagnosis of pancreatic tumors. MATERIALS AND METHODS: Contrast-enhanced ultrasonography with Levovist was performed on 62 consecutive patients (53 with pancreatic cancer, 4 with islet cell tumor, 3 with inflammatory pancreatic tumor, and 2 with metastatic tumor). The vascular and perfusion image phases of the tumors were evaluated and compared with the findings of contrast-enhanced computed tomography. RESULTS: Contrast-enhanced ultrasonography showed tumor vessels around and/or in the tumor at the vascular image phase in 79% of pancreatic cancer patients (42/53). At the perfusion image phase, 96% of pancreatic cancers (51/53) were classified as hypo-enhancement type. However, tiny spotty or irregular heterogeneous enhanced lesions were found in 84% of hypo-enhanced pancreatic cancer patients (43/51). The presence of small vessels at the vascular image phase was closely correlated with the presence of these intratumor regional enhanced lesions at the perfusion image phase (kappa coefficient=0.42). The sensitivity of contrast-enhanced ultrasonography (100%) for pancreatic cancer was superior to that of contrast-enhanced computed tomography (91%), but no significant difference was observed between the two (McNemar test: p=0.063). CONCLUSION: Contrast-enhanced ultrasonography with Levovist successfully visualizes fine vessels and enhancement in pancreatic tumors, and is useful for evaluating pancreatic tumors.