Multiplex Polymerase Chain Reaction Assay for Early Diagnosis of Viral Infection

Viral reactivation is one of the most serious complications for immunocompromised patients. Under immunosuppressive conditions, some viruses can be reactivated solely or simultaneously and may thus cause life-threatening infection. Therefore, the prompt and proper diagnosis of viral reactivation is important for the initiation of preemptive therapy. For this purpose, we recently developed a multiplex-virus polymerase chain reaction (PCR) assay. The multiplex PCR assay is designed to qualitatively measure the genomic DNA of 12 viruses at once: cytomegalovirus (CMV), human herpesvirus type 6 (HHV-6), HHV-7, HHV-8, Epstein-Barr virus (EBV), varicella-zoster virus (VZV), BK virus (BKV), JC virus (JCV), parvovirus B19 (ParvoB19), herpes simplex virus type 1 (HSV-1), HSV-2, and hepatitis B virus (HBV). When a specific PCR signal is obtained, the viral load is determined by a quantitative real-time PCR. The qualitative multiplex and quantitative real-time PCR procedures take only 3 hours to complete. With this assay system, we can identify viremia at the early stage and thereby prevent it from progressing to overt and symptomatic viral infection in immunocompromised patients, such as those receiving hematopoietic stem cell transplantation

トクシマシ シロヤマ ノ ホルトノキ ノ スイジャク コシ ノ ゲンイン ニツイテ : ホルトノキ イオウビョウ オ ヒキオコス ファイトプラズマ ノ シンコク ナ カンセン ジョウキョウ

Elaeocarpus sylvestris var. ellipticus was a dominant tree species in Mt. Shiroyama (castle mountain) in the Tokushima Central Park until 1970s, however, most of the E. sylvestris trees have died. In this study we investigated whether the death has been caused by Elaeocarpus yellows. Results of nested PCR revealed that DNA of phytoplasma, the pathogen of Elaeocarpus yellows, was found in all E. sylvestris trees in Mt. Shiroyama and about 80% E. sylvestris trees in the Tokushima Central Park. Results of PCR-RFLP showed that the all the trees had the same DNA type of phytoplasma, and it was identical to the one that had been found in Japan. These results indicated that the recent decrease of E. sylvestris in Mt. Shiroyama is due to Elaeocarpus yellows

トクシマシ シロヤマ ノ ホルトノキ ノ スイジャク コシ ノ ゲンイン ニツイテ : ホルトノキ イオウビョウ オ ヒキオコス ファイトプラズマ ノ シンコク ナ カンセン ジョウキョウ

Elaeocarpus sylvestris var. ellipticus was a dominant tree species in Mt. Shiroyama (castle mountain) in the Tokushima Central Park until 1970s, however, most of the E. sylvestris trees have died. In this study we investigated whether the death has been caused by Elaeocarpus yellows. Results of nested PCR revealed that DNA of phytoplasma, the pathogen of Elaeocarpus yellows, was found in all E. sylvestris trees in Mt. Shiroyama and about 80% E. sylvestris trees in the Tokushima Central Park. Results of PCR-RFLP showed that the all the trees had the same DNA type of phytoplasma, and it was identical to the one that had been found in Japan. These results indicated that the recent decrease of E. sylvestris in Mt. Shiroyama is due to Elaeocarpus yellows

Detection and management of cardiomyopathy in female dystrophinopathy carriers

Regular health checkups for mothers of patients with Duchenne muscular dystrophy have been performed at National Hospital Organization Tokushima Hospital since 1994. Among 43 mothers participated in this study, 28 dystrophinopathy carriers were identified. Skeletal and cardiac muscle functions of these subjects were examined. High serum creatine kinase was found in 23 subjects (82.1%). Obvious muscle weakness was present in 5 (17.8%) and had progressed from 1994 to 2015. Cardiomyopathy was observed in 15 subjects (60.0%), including dilated cardiomyopathy-like damage that was more common in the left ventricular (LV) posterior wall. Late gadolinium enhancement on cardiac MRI was found in 5 of 6 subjects, suggesting fibrotic cardiac muscle. In speckle tracking echocardiography performed seven years later, global longitudinal strain was decreased in these subjects, indicating LV myocardial contractile abnormality. These results suggest that female dystrophinopathy carriers should receive regular checkups for detection and treatment of cardiomyopathy, even if they have no cardiac symptoms

Cross-talk between TLR3 and TNF-α or IFN-γ Signaling in Induction of CXCL8/IL-8 and CXCL10/IP-10 Expression in Airway Epithelial Cells

CXCL8/IL-8 is a chemoattractant for neutrophils and mast cells, and regulates inflammatory cell recruitment in allergy, infection, and other neutrophil-related diseases. Interferon (IFN) -γ-inducible protein 10 (CXCL10/IP-10) is a chemokine that attracts mononuclear cells, Th1 cells, and natural killer cells. We investigated the levels of CXCL8/IL-8 and CXCL10/IP-10 expression by airway epithelial cells after exposure to the inflammatory cytokines tumor necrosis factor (TNF) -α and IFN-γ, and to poly I:C, a synthetic analog of double-stranded RNA that is a ligand of Toll-like receptor 3 (TLR3). Poly I:C, TNF-α, IFN-γ, and combinations of poly I:C with TNF-α or IFN-γ were used to stimulate the airway epithelial cell line BEAS-2B. Following stimulation, we determined CXCL8/IL-8 and CXCL10/IP-10 mRNA levels by real-time PCR and protein levels by ELISA. Poly I:C treatment upregulated mRNA and protein expression for both CXCL8/IL-8 and CXCL10/IP-10. The addition of TNF-α, but not IFN-γ, to poly I:C further increased the expression of CXCL8/IL-8 mRNA and protein. The addition of either TNF-α or IFN-γ to the poly I:C treatment further increased CXCL10/IP-10 mRNA and protein expression. Cross-talk between TLR3 signaling and inflammatory cytokines regulates the expression of CXCL8/IL-8 and CXCL10/IP-10 in airway epithelial cells. From our results, TNF-α and IFN-γ produce different effects on TLR3 signaling