Analysis of the regional anatomy of the retroesophageal right subclavian artery and surrounding structures

The retroesophageal right subclavian artery (RRSA) is a congenital anomalous branching of the arch of the aorta. Because its incidence is very low, it has not been fully understood how the RRSA develops during embryogenesis, and thus accumulation of observed findings in newly found cases is important to elucidate the etiology of the RRSA. We encountered a case of the RRSA during the course of gross anatomy dissection for medical students. The main findings in the present observations are that (a) the RRSA arose from the right side wall of the arch of the aorta as its last branch; (b) the detected RRSA was directed to the right and upward between the esophagus and vertebral column; (c) the right vertebral artery branched from the RRSA and entered the sixth cervical foramen transversarium; (d) the suprema intercostal artery branched from the costocervical trunk on both sides and its distal branches were distributed to the first and second intercostal spaces; and (e) both sides of bronchial arteries originated from the thoracic aorta. The present study gives further information about the morphological details of the RRSA leading to better understanding of its developmental process.

Generation and characterization of an antagonistic monoclonal antibody against an extracellular domain of mouse DP2 (CRTH2/GPR44) receptors for prostaglandin D2

Prostaglandin D2 (PGD2) is a lipid mediator involved in sleep regulation and inflammation. PGD2 interacts with 2 types of G protein-coupled receptors, DP1 and DP2/CRTH2 (chemoattractant receptor homologous molecule expressed on T helper type 2 cells)/GPR44 to show a variety of biological effects. DP1 activation leads to Gs-mediated elevation of the intracellular cAMP level, whereas activation of DP2 decreases this level via the Gi pathway; and it also induces G protein-independent, arrestin-mediated cellular responses. Activation of DP2 by PGD2 causes the progression of inflammation via the recruitment of lymphocytes by enhancing the production of Th2-cytokines. Here we developed monoclonal antibodies (MAbs) against the extracellular domain of mouse DP2 by immunization of DP2-null mutant mice with DP2-overexpressing BAF3, murine interleukin-3 dependent pro-B cells, to reduce the generation of antibodies against the host cells by immunization of mice. Moreover, we immunized DP2-KO mice to prevent immunological tolerance to mDP2 protein. After cell ELISA, immunocytochemical, and Western blot analyses, we successfully obtained a novel monoclonal antibody, MAb-1D8, that specifically recognized native mouse DP2, but neither human DP2 nor denatured mouse DP2, by binding to a particular 3D receptor conformation formed by the N-terminus and extracellular loop 1, 2, and 3 of DP2. This antibody inhibited the binding of 0.5 nM [3H]PGD2 to mouse DP2 (IC50 = 46.3 ± 18.6 nM), showed antagonistic activity toward 15(R)-15-methyl PGD2-induced inhibition of 300 nM forskolin-activated cAMP production (IC50 = 16.9 ± 2.6 nM), and gave positive results for immunohistochemical staining of DP2-expressing CD4+ Th2 lymphocytes that had accumulated in the kidney of unilateral ureteral obstruction model mice. This monoclonal antibody will be very useful for in vitro and in vivo studies on DP2-mediated diseases

Non-Welding Joint in Steel Plate Reinforced Concrete Structure : Part 1: Detail of Steel Plate Joint

日本建築学会大会学術講演梗概集（九州）2007年8

Novel method for immunofluorescence staining of mammalian eggs using non-contact alternating-current electric-field mixing of microdroplets

Recently, a new technique was developed for non-catalytically mixing microdroplets. In this method, an alternating-current (AC) electric field is used to promote the antigen-antibody reaction within the microdroplet. Previously, this technique has only been applied to histological examinations of flat structures, such as surgical specimens. In this study, we applied this technique for the first time to immunofluorescence staining of three-dimensional structures, specifically, mammalian eggs. We diluted an antibody against microtubules from 1:1,000 to 1:16,000, and compared the chromatic degree and extent of fading across dilutions. In addition, we varied the frequency of AC electricfield mixing from 5 Hz to 46 Hz and evaluated the effect on microtubule staining. Microtubules were more strongly stained after AC electric-field mixing for only 5 minutes, even when the concentration of primary antibody was 10 times lower than in conventional methods. AC electric-field mixing also alleviated microtubule fading. At all frequencies tested, AC electric-field mixing resulted in stronger microtubule staining than in controls. There was no clear difference in a microtubule staining between frequencies. These results suggest that the novel method could reduce antibody consumption and shorten immunofluorescence staining time

Axillary schwannoma mimicking lymph node metastasis-associated breast cancer: a case report

Abstract Background Axillary schwannoma associated with breast cancer is an extremely rare disease, and previous reports have been limited. In this setting, there is great concern about whether a tumor in the axillary region is lymph node metastasis. Herein, we report a unique case of axillary schwannoma that mimicked lymph node metastasis associated with breast cancer. Case presentation A 68-year-old woman who underwent mastectomy and axillary lymph node dissection for right breast cancer over 20 years ago presented to our hospital with numbness and weakness in the right arm for 6 months. Ultrasonography, computed tomography, and magnetic resonance imaging showed a 20-mm well-circumscribed round tumor in the right axillary region. Initially, she was suspected of having lymph node metastasis-associated breast cancer, but the result of the core needle biopsy was a schwannoma. The patient underwent tumor enucleation. The patient has had no recurrence 1 year after the operation. Conclusion Axillary schwannomas often mimic lymph node metastasis in patients with a history of malignancy, particularly breast cancer. To select the optimal treatment, the clinicians should make as accurately as possible a diagnosis, with histopathological examinations, when examining patients with cancer who develop tumors in the axillary region

Cyclic di-GMP Signaling Regulates Invasion by Ehrlichia chaffeensis of Human Monocytes ▿ †

Cyclic di-GMP (c-di-GMP) is a bacterial second messenger produced by GGDEF domain-containing proteins. The genome of Ehrlichia chaffeensis, an obligatory intracellular bacterium that causes human monocytic ehrlichiosis, encodes a single protein that contains a GGDEF domain, called PleD. In this study, we investigated the effects of c-di-GMP signaling on E. chaffeensis infection of the human monocytic cell line THP-1. Recombinant E. chaffeensis PleD showed diguanylate cyclase activity as it generated c-di-GMP in vitro. Because c-di-GMP is not cell permeable, the c-di-GMP hydrophobic analog 2′-O-di(tert-butyldimethylsilyl)-c-di-GMP (CDGA) was used to examine intracellular c-di-GMP signaling. CDGA activity was first tested with Salmonella enterica serovar Typhimurium. CDGA inhibited well-defined c-di-GMP-regulated phenomena, including cellulose synthesis, clumping, and upregulation of csgD and adrA mRNA, indicating that CDGA acts as an antagonist in c-di-GMP signaling. [32P]c-di-GMP bound several E. chaffeensis native proteins and two E. chaffeensis recombinant I-site proteins, and this binding was blocked by CDGA. Although pretreatment of E. chaffeensis with CDGA did not reduce bacterial binding to THP-1 cells, bacterial internalization was reduced. CDGA facilitated protease-dependent degradation of particular, but not all, bacterial surface-exposed proteins, including TRP120, which is associated with bacterial internalization. Indeed, the serine protease HtrA was detected on the surface of E. chaffeensis, and TRP120 was degraded by treatment of E. chaffeensis with recombinant E. chaffeensis HtrA. Furthermore, anti-HtrA inhibited CDGA-induced TRP120 degradation. Our results suggest that E. chaffeensis invasion is regulated by c-di-GMP signaling, which stabilizes some bacterial surface-exposed proteins against proteases