Objective To analyze the full length cDNA sequence of Schistosoma japonicum egg miracidia genes harboring signal sequence.Methods The gene specific primers were designed and synthesized according to S.japonicum egg miracidia cDNA fragment containing signal sequence identified by signal sequence trapping method previously. The 5′and 3′ end cDNA fragments of each egg miracidia cDNA fragment harboring signal sequence were amplified by nest PCR using the first strand cDNA of S.japonicum as the template. The specific PCR fragments were cloned by TA clone method and sequenced. The full length cDNA sequence of each gene with signal sequence was constructed by comparing the cDNA sequence identified with signal sequence trapping method and the 5′ end sequence, the 3′ end sequence and deleting the overlapping fragments. The splicing model between mRNA of signal sequence and one of mature portions of S.japonicum egg miracidia gene was checked by analyzing the genomic DNA sequence structure of some genes with signal sequence. Results The 5′ and 3′ end cDNA fragments of sixteen among thirty cDNA fragment with signal sequence were amplified successfully, and their DNA sequences were determined. The full length cDNA sequences of sixteen egg miracidia genes were obtained by sequence matching and splicing. The results of deduced amino acid analysis found that the signal peptide of gene SjP4001 was the same to the one of SjP1531 and the signal peptide of gene SjP1183 was similar to the one of gene SjP3742. It confirmed that different genes could share the same or similar signal peptide. The data of S.japonicum genomic DNA sequence analysis showed that the S.japonicum could obtain its signal sequence by alternative splicing model or trans-splicing model. Conclusions The full length cDNA sequences of sixteen S.japonicum egg miracidia genes with signal sequence have been defined, it indicated that the S.japonicum egg miracidia genes could get its signal sequence by alternative splicing model or trans-splicing model was found in this study.

Nineteen stocks of Trypanosoma cruzi originating from several endemic countries for Chagas‘ disease in Central and South America were subjected to two-dimensional protein electrophoresis analysis. The presence or absence of a total of492polypeptide spots among19gel profiles was determined. The stocks were classified into three major distinctive groups derived from (I) Central America and the northern part of South America; (IIa) Central America and the northern part of South America; and (IIb) central and southern parts of South America, which showed perfect concordance with the previously reported classification based on isozyme and DNA sequence analyses. Late log phase of each epimastigote was inoculated to human cell lines WI-38and Hs224.T originating from the lung and muscle, respectively, and the number of trypomastigotes released was counted. The number of trypomastigotes from T. cruzi in group I released from the two cell lines was significantly higher than that in group III (p&It;0.05). The findings suggested that the phenetic distance appearing within the T. cruzi may, to some extent, be associated with the intracellular growth of T. cruzi, one of the characteristic features of growth found in the species.

We perfomed an aortic valve replacement and intraoperative plasma exchange with Cell Saver 4^® made by Haemonetics for removal of bilirubin. Diluted blood after cardiopulmonary bypass and suctioned blood (total volume 11, 300ml) was washed, concentrated and transfused. Plasma bilirubin level was decreased to 5.4mg/dl from 9.9mg/dl during operation. It was concluded that intraoperative plasma excange with Cell Saver 4^® was safe and effective technique for removal of bilirubin during open heart surgery.

An internal felt-reinforced patch-plasty was performed in 11 patients with dissecting aortic aneurysm (DeBakey type I: 4 cases, type II: 1 case, type III: 5 cases, aortic arch dissection: 1 case). The aortic cross-clamp time was 84±19 min on the average. The initial tear of the aortic intima was closed on 10 patients. Minor leakage through a felt inserted in the false lumen was observed in one patient of type I. There was no operation-related death except one patient of type III who died from arrythmia encountered following termination of centrifugal pump bypass. Thrombotic occlusion of the false lumen developed in the ascending aorta in type I and II cases, and in the desceding aorta in type III one month after operation. The false lumen localized in the aortic arch was completely occluded by thromi. Postoperative course was excellent in all patients after 16 months on the average. Internal felt-reinforced patch-plasty is a simple and reliable procedure for closing the intial tear of dissecting aortic aneurysms.

The aim of this study is to collect the suggestions and recommendations of health care professionals involved in the regional palliative care program (OPTIM-study). A total of 101 multidisciplinary health care professionals who participated in the intervention program were interviewed, and 107 meaningful units were obtained from 89 valid interviews. The responses were categorized into “suggestions regardless of profession” (n=59), including “Participate in a multi-disciplinary conference to expand the network of people”, “Try to understand the situation of others”, “Seek support from others when you cannot solve the problem by yourself”, “Pursue all possibilities before giving up”, and “Do not try too hard”. As suggestions to those engaged in each profession, “Suggestions to community pharmacies” and “Suggestions to care managers” are frequently described. An examination of suggestions by the participants in the regional palliative care program could provide some insights to improve community palliative care.

The purpose of this study was to identify the greatest impact of the regional palliative care program on community health care professionals. Interviews were conducted involving 101 people who became involved in the intervention program implemented in 4 areas across Japan, and 96 valid responses were collected. The following were cited as the greatest impact: [I developed a network of people, and realized the importance of collaboration] (n＝61; “I was able to develop an interpersonal relationship” and “Now I understand the significance of collaboration”), [My knowledge and skills regarding palliative care were improved] (n＝18; “Knowledge and support helped me respond to patients with confidence” and “I have come to think that there is more to palliative care than terminal care”), [I rediscovered my role through a wide variety of experiences] (n＝10), [Both collaboration and palliative knowledge/skills meant a lot to me] (n＝4), [What I experienced in this program will help me play my role] (n＝2), and [Patients and their families became satisfied] (n＝1). The community palliative care program was most effective in facilitating collaboration, and helped participants develop knowledge and skills concerning palliative care.

In malaria endemic areas, people naturally acquire an age-related immunity to malaria. Part of this immunity involves anti-malarial specific antibodies. Acquisition of these malaria-specific antibodies depends not only on exposure to malaria parasites but also on the human genetic predisposition. CTLA-4 is a costimulatory molecule that delivers an inhibitory signal to suppress T-cell as well as B-cell responses. We investigated associations between malaria-specific antibody levels and CTLA-4 polymorphisms in 189 subjects living in a hyper-endemic area of Papua New Guinea (PNG), where both P. falciparum and P. vivax are prevalent. We determined P. falciparum⁄ P. vivax specific IgG⁄IgE levels (Pf-IgG, Pv-IgG, Pf-IgE, Pv-IgE) and polymorphisms in the CTLA-4 gene at position -1661 promoter region (A⁄G), the +49 exon 1 non-synonymous mutation (A⁄G), and the +6230 3‘-UTR (A⁄G). All quantified antibody levels were significantly higher in subjects > 5 years (n = 150) than in subjects ≤ 5 years of age (n = 39). In children ≤ 5 years old, significant associations were detected between CTLA-4 +49 (GG⁄AG vs. AA) and Pv-IgG (median 18.7 vs. 13.7 Μg⁄ml, P = 0.017) and Pv-IgE (266.6 vs. 146.5 pg⁄ml, P = 0.046). No significant difference was observed in subjects > 5 years old. These results suggest that the CTLA-4+49 polymorphism influenced Pv-IgG and Pv-IgE levels among children less than five years old in the studied population, which may regulate the age- and species-specific clinical outcomes of malaria infection.

Immunoepidemiological studies from endemic areas have revealed age-dependent resistance correlation with increased level of IgE and decreased level of IgG4 antibodies in responses to schistosomes’ soluble worm antigen. However, there have been limited studies on analyses of major antigens that provoke IgE and IgG4 immune response during chronic stage of schistosomiasis. In this study, for the first time, immunoproteomics approach has been applied to identify S. japonicum worm antigens in liquid fractions that are recognized by IgE and IgG4 antibody using plasma from chronically infected population. ProteomeLabPF 2D fractionated 1-D and 2-D fractions of SWA antigens were screened using pooled high IgE/IgG4 reactive plasma samples by dot-blot technique. In 1-D fractions, IgE isotype was detected by fewer antigenic fractions (43.2%). The most recognized isotype was IgG3 (79.5%) followed by IgG1 (75.0%) and IgG4 (61.4%). Liquid chromatography MS/MS protein sequencing of reactive 2-D fractions revealed 18 proteins that were identified, characterized and gene ontology categories determined. 2-D fractions containing proteins such as zinc finger, RanBP2-type, domain-containing protein were strongly recognized by IgE and moderately by IgG4 whereas fractions containing proteins such as ubiquitin-conjugating enzyme and cytosolic II 5'-nucleotidase strongly recognizing by IgG subclasses (IgG1, IgG3 and IgG4) but not IgE. By this study, a simple and reproducible proteomic method has been established to identify major immunoreactive S. japonicum antigens. It is anticipated that this will stimulate further research on the immunogenicity and protective potential of proteins identified as well as discovery of novel compounds that have therapeutic importance.