Immunogold electron microscopic evidence of in situ formation of homo- and heteromeric purinergic adenosine A1 and P2Y2 receptors in rat brain

<p>Abstract</p> <p>Background</p> <p>Purines such as adenosine and ATP are now generally recognized as the regulators of many physiological functions, such as neurotransmission, pain, cardiac function, and immune responses. Purines exert their functions via purinergic receptors, which are divided into adenosine and P2 receptors. Recently, we demonstrated that the G_i/o-coupled adenosine A₁receptor (A₁R) and G_q/11-coupled P2Y₂receptor (P2Y₂R) form a heteromeric complex with unique pharmacology in co-transfected human embryonic kidney cells (HEK293T). However, the heteromeric interaction of A₁R and P2Y₂R <it>in situ </it>in brain is still largely unknown.</p> <p>Findings</p> <p>In the present study, we visualized the surface expression and co-localization of A₁R and P2Y₂R in both transfected HEK293T cells and in rat brain by confocal microscopy and more precisely by immunogold electron microscopy. Immunogold electron microscopy showed the evidence for the existence of homo- and hetero-dimers among A₁R and P2Y₂R at the neurons in cortex, cerebellum, and particularly cerebellar Purkinje cells, also supported by co-immunoprecipitation study.</p> <p>Conclusion</p> <p>The results suggest that evidence for the existence of homo- and hetero-dimers of A₁R and P2Y₂R, not only in co-transfected cultured cells, but also <it>in situ </it>on the surface of neurons in various brain regions. While the homo-dimerization ratios displayed similar patterns in all three regions, the rates of hetero-dimerization were prominent in hippocampal pyramidal cells among the three regions.</p

Effectiveness of Cross‐Linked Gelatin Glue in Canine Lung Surgery Models.

Background. Air leakage is a common postoperative complication in pulmonary surgery, and surgical sealants have been developed to prevent or reduce the incidence of air leaks. In this study, we evaluated the efficacy of cross-linked gelatin glue (gelatin plus glutaraldehyde) in canine lung surgery models. Methods. Pulmonary fistulas and injuries were created in dogs and sealed with gelatin glue, fibrin glue, or fibrin glue with a polyglycolic acid (PGA) sheet. Seal-breaking pressures were measured in the fistula model, and pleural adhesions were assessed 28 days postoperatively in the lung injury model. Results. The seal-breaking pressures for canine cadaver and living lung surgeries (; the maximum pressures were 80 and 40 cm H₂O) were respectively: gelatin glue, 77 ± 6 and 32.3 ± 8.9cm H₂O; fibrin glue using spray, 39.2 ± 9.3 and 32 ± 6cm H₂O; fibrin glue using the rub-and-soak method, 35 ± 13.4 and 40 ± 0 cm H₂O; and fibrin glue with a PGA sheet, 55.5 ± 18.2 and 39 ± 2cm H₂O. In the lung injury model, there were no chest wall adhesions in the gelatin and fibrin glue alone groups, while strong adhesions were observed when treated with fibrin glue with a PGA sheet. Conclusions. Gelatin glue's sealing effect was superior to that of fibrin glue while preventing postoperative pleural adhesions. These findings suggest that gelatin glue may be effective as a surgical sealant or anti-adhesion materialin lung surgery

Evaluation of Radiation Exposure Literacy among Mammography Examinees Using Radiation Dose Distribution in Mammography Examinations

東京都立大学Tokyo Metropolitan University博士（放射線学）doctoral thesi

Clinical Usefulness of Multiplex PCR Lateral Flow in MRSA Detection: A Novel, Rapid Genetic Testing Method

Methicillin-resistant Staphylococcus aureus (MRSA) with exogenous cassette DNA containing the methicillin-resistant gene mecA (SCCmec) poses a problem as a drug-resistant bacterium responsible for hospital- and community-acquired infections. The frequency of MRSA detection has recently been increasing rapidly in Japan, and SCCmec has also been classified more diversely into types I–V. A rapid test is essential for early diagnosis and treatment of MRSA infections, but detection by conventional methods requires at least two days. The newly developed multiplex PCR lateral flow method allows specific amplification of femA to detect S. aureus, mecA to detect SCCmec, and kdpC to detect SCCmec type II; moreover, PCR products can be evaluated visually in about 3 h. In the present study, we developed a PCR lateral flow method for MRSA using this method and investigated its clinical usefulness in the detection of MRSA. The results showed a diagnostic concordance rate of 91.7% for MRSA and methicillin-susceptible S. aureus between bacteriological examination and PCR lateral flow, and a high level of specificity in PCR lateral flow. In addition, a higher detection rate for S. aureus using the same sample was observed for PCR lateral flow (70.2%) than for bacteriological tests (48.6%). The above results show that PCR lateral flow for MRSA detection has high sensitivity, specificity, and speed, and its clinical application as a method for early diagnosis of MRSA infections appears to be feasible