Calmodulin and cyclic ADP-ribose interaction in Ca2+ signaling related to cardiac sarcoplasmic reticulum: superoxide anion radical-triggered Ca2+ release

Reactive oxygen species (ROS) are often shown to damage cellular functions. The targets of oxidative damage depend on the nature of ROS produced and the site of generation. In contrast, ROS can also regulate signal transduction. In this case, ROS may either induce or enhance events, which lead to forward directions of cellular signaling. The consequences of regulation of signal transduction can be observed in physiological processes such as muscle contraction. Here, we discuss the concentration-dependent effects of superoxide anion radical (*O2-) on Ca2+ release from the cardiac sarcoplasmic reticulum (SR). Recent studies suggest that the ADP-ribosyl cyclase pathway, through its production of cyclic adenosine 5'-diphosphoribose (cADPR), may control Ca2+ mobilization in cardiac muscle cells. *O2- has dual effects that are concentration dependent. At low concentrations (nearly nanomolar levels), *O2- induces Ca2+ release by stimulating synthesis of cADPR, which requires calmodulin for sensitization of ryanodine-sensitive Ca2+-release channels (RyRC). At these low concentrations, *O2- is responsible for regulation of cellular signal transduction. At higher concentrations (micromolar levels), *O2- produces a loss in the function of calmodulin that is to inhibit RyRC. This results in an increase in Ca2+ release, which is linked to cell injury. The difference in the functions of low and high concentrations of *O2- may result in two distinct physiological roles in cardiac muscle Ca2+ signaling.     

Evidence of the monoclonal composition of human endometrial epithelial glands and mosaic pattern of clonal distribution in luminal epithelium

The endometrium is a highly regenerative tissue that plays a crucial role in implantation. We examined the clonal constitution of glandular cells as well as the luminal epithelium of this unique tissue. Using collagenase-based digestion techniques with microscopic manipulation, we isolated individual human endometrial glands and examined their clonality using a polymerase chain reaction-based assay for nonrandom X chromosome inactivation with an X-linked androgen receptor gene. Most of the glands analyzed were composed of monoclonal populations of epithelial cells and one of the glands exhibited a loss of heterogeneity in the androgen receptor gene. In addition, adjacent glands within a 1-mm(2) area shared clonality, suggesting that clonality of the luminal epithelium is regionally defined. The clonality of endometrium was further confirmed in a study of female mice that harbor the green fluorescent protein gene on either the maternal or paternal X chromosome. Fluorescent microscopy of uterine sections revealed that individual endometrial glands consisted completely of either fluorescent or nonfluorescent cells and that the surface epithelium exhibited a clear boundary between these cell types. These findings suggest that single or multiple stem cells with uniform clonality exist on the bottom of each endometrial gland and genetic alterations occurring in such cells may play a critical role in endometrial carcinogenesis. The possible association between area-specific X inactivation of the endometrial surface and the endometrial receptivity of embryo implantation remains to be clarified.     

Histopathological explanation of the MRI target sign in extra-axial schwannomas

BackgroundTo better understand the nature of magnetic resonance imaging (MRI) findings in schwannomas, especially in the “target sign” of these findings, the histopathological investigation was performed.MethodsThe MRI findings were correlated with histopathological features in 22 samples of schwannomas, which were mostly resected from the extremities. The histopathological analyses included alcian blue staining and immunohistochemical staining for S-100 protein, proliferating cell nuclear antigen (PCNA) and epithelial membrane antigen (EMA).ResultsSeven of the 22 samples of schwannomas of the extremities exhibited target signs including a peripheral zone of homogeneously high signal intensity and a central zone of heterogeneous signal intensity in T2-weighted images. Gadolinium-enhanced T1–weighted images demonstrated a central heterogeneous enhancement and a peripheral ring of homogeneously low signal intensity. Histopathologically, S-100 and PCNA were positive only in the central heterogeneous signal area. In contrast, EMA was only stained on the degenerative epi/perineurium in the peripheral zone.ConclusionIn schwannomas of the extremities showing target sign in T2-weighted images, histopathologically, the peripheral areas were suggested to be mucinous degeneration of the epineurium or perineurium, while the central areas were composed of truly neoplastic cells.     

Anaplastic large cell Ki-1 lymphoma with bone involvement: Report of two cases

Two cases of anaplastic large cell Ki-1 lymphoma involving bone as the most prominent and initial manifestation are reported. The first patient was a 20-year-old male who had back pain and incomplete paraparesis due to vertebral involvement. The second was a 14-year-old girl, whose first clinical signs were fever of unknown origin and sternal bone pain. Radiologically, skeletal lesions were lytic and destructive. Histopathologically, the tumour cells had pleomorphic bizarre nuclei and abundant basophilic cytoplasm. Immunohistochemically, Ki-1 (CD30) reactivity was strongly positive in both cases. Tumour cells were also CD3, CD4, epithelial membrane antigen and interleukin-2 receptor positive in the first case, and CD10, HLA-DR positive in the second case. The former tumour was considered to be of T-cell lineage and the latter of lymphoid progenitor cell origin. Radiation and chemotherapy were temporarily effective. However, both patients died 14 and 7 months after diagnosis, respectively, due to systemic lymph node involvement. These observations suggest that the prognosis for Ki-1 lymphoma involving bone is poorer than indicated in previous reports.     

Dynamic mechanical properties of dental amalgams

The dynamic mechanical properties of two high-copper amalgams (Tytin and Dispersalloy) and two traditional amalgams (Aristalloy and Aristalloy with Zn) were measured over a temperature range of 0-70 degrees C and at frequencies of 0.1, 1, and 10 Hz by means of a DuPont DMA. Values of storage modulus (E') for the amalgams were equivalent to the Young's modulus (E) measured from static mechanical test methods, with Dispersalloy demonstrating the highest moduli. Values of E' decreased with increased temperature. E' of traditional amalgams decreased more rapidly than did those of the Cu-rich amalgams. Values of loss modulus (E") for Tytin were smaller than those of Dispersalloy and the two types of Aristalloy. High values of E" for the traditional amalgams correspond to a greater viscous behavior. Marked differences between the magnitude of tan delta and its temperature coefficients for traditional and high-copper amalgams were observed, which is indicative of differences in visco-elastic behavior between these two amalgam systems.     

Preparation of the antibodies against recombinant human thymidylate synthase for the detection of its intratumoral levels and the application to sensitivity-study of 5-fluorouracil

Thymidylate synthase (TS) is a rate-limiting enzyme in de novo DNA biosynthesis and also a primary target for 5-fluorouracil (5-FU)-based chemotherapy. To estimate the con-elation between the expression of TS in the tumor, clinical response and prognosis in cancer patients treated with 5-FU, we have prepared recombinant human TS (rhTS) protein by culturing of E. coli transfected with the plasmid (pGEX-2TH) encoding the nucleotide sequence of hTScDNA and then obtained a highly purified polyclonal antibody against rhTS protein. Using this anti-TS antibody, it was revealed that the content of TS protein, as determined by Western blot analysis, correlated with the enzyme activity (gamma=0.973) and cytotoxicity of 5-FU, expressed as IC50 value (gamma=0.954) against human colon tumor cells, sensitive to and with acquired-resistance to 5-fluoropyrimidines and other cancer cells. On immunochemical evaluation, it was also confirmed that the tumor cells overexpressing TS proteins were strongly stained by the polyclonal antibody when compared to the cell lines expressing TS to lower extent both in vitro and in vivo conditions. These results indicate that this purified polyclonal antibody to rhTS is applicable to prospective and retrospective clinical studies on immunochemical TS expression in various tumors as a prognostic factor and 5-FU response-predicting parameter.     

Accumulation of L-[2-(F-18)]fluorophenylalanine in peri-infarct area in a patient with acute cerebral infarction

We studied the brain uptake of amino acid in a patient with acute cerebral infarction with L-[2-(F-18)] fluorophenylalanine and positron emission tomography. The increased accumulation of the ligand was specifically found in the peri-infarct area where oxygen metabolism was still maintained but decreased later in the 72-day follow-up period. The kinetic analysis revealed that increased accumulation was not due to increased transport from the blood to the brain but to delayed washout from the brain to the blood. Although the mechanism is still unknown, abnormally high accumulation of L-[F-18]fluorophenylalanine may predict delayed neuronal changes after ischemic insults of the brain.