Kenji Masunaga

Clinical and laboratory features of fatal rapidly progressive interstitial lung disease associated with juvenile dermatomyositis

OBJECTIVE Rapidly progressive interstitial lung disease (RP-ILD) is a rare but potentially fatal complication of JDM. The aim of this study was to establish markers for the prediction and early diagnosis of RP-ILD associated with JDM. METHODS The clinical records of 54 patients with JDM were retrospectively reviewed: 10 had RP-ILD (7 died, 3 survived), 19 had chronic ILD and 24 were without ILD. Routine tests included a high-resolution CT (HRCT) scan of the chest and measurement of serum levels of creatine phosphokinase, ferritin and Krebs von den Lungen-6 (KL-6). Anti-melanoma differentiation-associated gene 5 (MDA5) antibodies and IL-18 levels were measured by ELISA. RESULTS No differences were found in the ratio of juvenile clinically amyopathic DM between the three groups. Initial chest HRCT scan findings were variable and could not distinguish between RP-ILD and chronic ILD. Anti-MDA5 antibodies were positive in all 8 patients with RP-ILD and 10 of 14 with chronic ILD, but none of the patients without ILD. Serum levels of anti-MDA5 antibody, ferritin, KL-6 and IL-18 were significantly higher in the RP-ILD group than in the chronic ILD and non-ILD groups. Serum levels of IL-18 positively correlated with serum KL-6 (R = 0.66, P < 0.001). CONCLUSION High serum levels of IL-18, KL-6, ferritin and anti-MDA5 antibodies (e.g. >200 units by ELISA) are associated with RP-ILD. These can be used as an indication for early intensive treatment. Both alveolar macrophages and autoimmunity to MDA5 are possibly involved in the development of RP-ILD associated with JDM.

Mapping of functional domains on the influenza A virus RNA polymerase PB2 molecule using monoclonal antibodies.

Summary. Monoclonal antibodies against the PB2 of A/Puerto Rico/8/34 (A/PR/ 8/34) (H1N1) were prepared in order to define the functional domains of the RNA polymerase of influenza virus. The fifteen monoclonal antibodies that were generated were divided into 4 groups on the basis of ELISA binding to PB2 or its peptide fragments. Six Group I antibodies that bound to the PB2 N-terminal region (amino acids 1–104) did not inhibit transcription by the viral ribonucleoprotein complex. A single Group II antibody recognizing the region of amino acids 206–259 inhibited ApG-primed transcription. Groups III and IV antibodies bound to the C-terminal region of amino acids 660–759. Of these, Group III antibodies inhibited transcription. The present results identify multiple monoclonal antibody binding domains in PB2, two of which, when bound by antibodies, negatively affect viral RNA transcription.

A pilot study on the changes in immunity after ACTH therapy in patients with West syndrome

Adrenocorticotropic hormone (ACTH) has been the first-line drug for the treatment of West syndrome, although the therapy has various adverse effects. ACTH depresses resistance to a variety of bacterial, viral, protozoal, and fungal agents. The timing of the various vaccinations is delayed after ACTH therapy in Japan, because the immune system is believed to be affected for approximately 6 months. However, the duration of the effect of ACTH on the immune system is not known. Therefore, we examined changes in the immunity levels before and after ACTH therapy. We measured white blood cell counts, lymphocyte counts, T/B cell counts, CD4(+) and CD8(+) T cell counts, CD 4/8 ratio, lymphocyte blastoid transformation by PHA or Con-A, and the levels of IgA, IgM, and IgG before, immediately after, and 1, 3, 6, and 12 months after ACTH therapy. The lymphocyte counts and CD4(+) T cell counts were significantly decreased immediately after and at 1 and 3 months after the therapy, and did not return to the previous levels even at 6 months and 12 months after ACTH treatment; however, these levels returned to within normal limits (within the 95% confidence interval). Immunoglobulin levels did not change after the ACTH therapy. Helper T cells were more depressed than cytotoxic T cells after ACTH therapy.

Potential Drug Interaction between Warfarin and Linezolid

OBJECTIVE Warfarin is known to interact with many drugs; however, there are currently no descriptions of an interaction with linezolid in the literature. It was recently brought to our attention, however, that several warfarin-medicated patients have experienced an increase in the prothrombin time international normalized ratio (PT-INR) following the administration of linezolid. We therefore performed a retrospective survey in order to investigate the possibility of an interaction between warfarin and linezolid. METHODS The survey items included age, gender, underlying disease, type of surgery, type of infectious disease, duration of linezolid administration, laboratory values and the dose of warfarin. The PT-INR was observed over time before treatment and at days 4 or 5 and 10, completion and one week after the end of concomitant therapy. Patients The subjects included six patients who were recovering from recent heart-related surgery. RESULTS The PT-INR increased from 1.62±0.32 before concomitant linezolid administration to 3.00±0.83 at day 4 or 5 after concomitant administration (p<0.01) and significantly decreased from 1.65±0.45 at the completion of the regimen to 1.26±0.1 one week later (p<0.05). With respect to the relationship between the dose of warfarin and the PT-INR in five cases, the PT-INR increased following concomitant linezolid treatment in all cases. CONCLUSION Although it has been reported that linezolid does not influence the metabolism or protein binding of warfarin, our data showed potential drug interactions between warfarin and linezolid. Our data suggest that PT-INR monitoring after the completion of concomitant warfarin and linezolid therapy is important.

Antibody-Scanning and Epitope-Tagging Methods Molecular Mapping of Proteins Using Antibodies

Because synthetic short peptides bearing critical binding residues, can chemically mimic the folded antigenic determinants on proteins, short synthetic peptides can generate antibodies that react with cognate sequences in intact folded proteins. According to this mimotope theory, we produced site-specific antibodies by immunization with short peptides which overlapped each other and covered the entire protein, and used them for domain mapping of influenza virus RNA polymerase (antibody-scanning method). We also used a tagged-epitope and its monoclonal antibodies for topology mapping of clathrin light chains in clathrin triskelions by electron microscopy. Both methods using specific epitopes in combination with their antibodies enable us to determine the domains of interesting proteins systematically without the need to generate monoclonal antibodies or mutant proteins.

Epitope mapping of the influenza A virus RNA polymerase PA using monoclonal antibodies.

Summary. To obtain reagents to functionally map the PA protein, we produced monoclonal antibodies specific to this protein. Twenty-two monoclonal antibodies reacting with PA protein in ELISA were divided into 10 groups on the basis of competitive binding patterns to this protein. Of these, seventeen monoclonal antibodies bound to PA polypeptide spanning amino acids 101–400 and three bound to that of amino acids 518–600, while the other two did not react with any PA polypeptides tested with the exception of the intact PA. Among these monoclonal antibodies, only five reacted with PA in A/PR/8/34 virus-infected cells in indirect immunofluorescence assay. Thus, we obtained monoclonal antibodies that recognize at least 10 distinct regions of the PA molecule. These monoclonal antibodies should be useful in dissecting functions of the PA protein.

Nucleic acid binding by zinc finger-like motif of human papillomavirus type 16 E7 oncoprotein

Human papillomavirus (HPV) types 16 and 6b E7 proteins and their chimeric or mutant proteins were analyzed for oligonucleotide-binding activity by surface plasmon resonance-based biomolecular interaction analysis. The results indicated that type 16 E7 protein has stronger nucleic acid-binding activity than that of type 6b E7 protein. In addition, the results also indicated that the zinc finger-like motif in the C-terminal region of the type 16 E7 protein plays an important role in this activity.

Pediatric Rheumatology Association of Japan recommendation for vaccination in pediatric rheumatic diseases

Abstract Pediatric Rheumatology Association of Japan has developed evidence-based guideline of vaccination in pediatric rheumatic diseases (PRDs) as a part of Guideline of Vaccination for Pediatric Immunocompromised Hosts. Available articles on vaccination in both adult rheumatic diseases and PRDs were analyzed. Non-live vaccines are generally safe and effective in patients with PRDs on corticosteroid, immunosuppressant, and/or biologics, although efficacy may be attenuated under high dose of the drugs. On the other hand, efficacy and safety of live-attenuated vaccine for the patients on such medication have not been established. Thus, live-attenuated vaccines should be withheld and, if indicated, may be considered as a clinical trial under the approval by Institutional Review Board. All patients with PRDs anticipating treatment with immunosuppressants or biologics should be screened for infection of hepatitis B and C and tuberculosis before the commencement of medication. Varicella vaccine should be considered in sensitive patients ideally 3 weeks or longer before the commencement of immunosuppressants, corticosteroids, or biologics. Bacille Calmette-Guérin should be withheld at least for 6 months after birth, if their mothers have received anti-tumor necrosis factor-α antibodies during the second or third trimester of pregnancy.

Characteristics of Multidrug-Resistant Corynebacterium spp. Isolated from Blood Cultures of Hospitalized Patients in Japan

Corynebacterium is a genus consisting of Gram-positive, rod-shaped bacteria, that is wildly distributed in nature. We report the epidemiological characterization of Corynebacterium spp. isolated from blood specimens at the Kurume University Hospital, between June 2008 and November 2011. Twenty-two strains that were likely Corynebacterium spp. were isolated from 22 hospitalized patients, of which 12 (54.5%) were identified as Corynebacterium striatum. Minimum inhibitory concentration tests were performed after biochemical and genotypic identifications. Biofilm production was detected using a 96-well microplate assay. The dissemination of C. striatum was investigated using pulsed-field gel electrophoresis (PFGE). All strains showed the tendency to be resistant to multiple drugs except vancomycin. Fourteen (82.4%) strains, including 9 C. striatum strains were capable of producing biofilms. Four distinct PFGE patterns were detected among C. striatum strains; 6 of which were identified as dominant pattern A (defined in this study) and had high biofilm production ability. During the 3-year monitoring period, these strains might have repeatedly infected the patients or could have readily colonized the hospital environments. C. striatum appeared to be a potential risk factor for bloodstream infections in hospitalized patients. More surveillance and enhanced control strategies are necessary to decrease Corynebacterium spp. infections in hospitals.

Post-Travel Consultations in a Regional Hub City Hospital, Japan

OBJECTIVE To clarify the characteristics of post-travel consultation services in Japan, particularly in the provinces, we analyzed our post-travel patients in the travel clinic of Kurume University Hospital located in Kurume City (a regional hub City in southwestern Japan). METHODS Sixty post-travel patients visited our clinic between April 2008 and October 2014 and participated in this study: 55 were Japanese and five were foreign. We summarized and compared the characteristics of the patients after dividing the Japanese participants into long-term travelers (>14 days) and short-term travelers (≤14 days). The foreign travelers were described in a separate analysis. RESULTS Of the 55 Japanese travelers, the mean age (± standard deviation) was 37.3 ± 16.3 years, and 36 patients (65%) were men. Southeast Asia was the major destination (30/55, 55%), and business was stated as the major reason for travel (16/55, 29%). Post-exposure rabies prophylaxis (16/55, 29%) was the most common purpose for the consultations. There were 34 participants (62%) who were classified as short-term travelers. Fewer of the short-term travelers stated receiving pre-travel consultations compared with long-term travelers (11% vs. 79%, p=0.0002). The five foreign travelers included one dengue fever patient and two malaria patients. CONCLUSION Most post-travel Japanese patients visited our clinic were short-term travelers who had not received any pre-travel consultation. One of the most common complaints, post-exposure rabies prophylaxis, could have been avoided to some extent by appropriate pre-travel consultations. The results of this study suggest that pre-travel consultations should therefore be encouraged for both long- and short-term travelers.