Phosphoinositide-specific phospholipase C(PLC) is known as a key enzyme which produces two major second messengers: diacylglycerol and inositol 1,4,5 trisphosphate. Although it has been suggested that PLC beta isozymes have important roles in nervous system, less is known about the function of PLC beta in development of nervous system. We have localized the mRNA expressions of PLC beta isozymes in the postnatal rat brains by id firm hybridization histochemistry. In the postnatal rat brains, each isozyme of PLC beta showed differential expression pattern. The expression of PLC beta1 mRNA was found in various areas including olfactory bulb, cerebral cortex, caudate putamen, hippocampus, dentate gyrus, and cerebellum. In general, the expression in these areas was gradually increased after birth (PO) until postnatal day 21 (P2l) and slightly decreased to adult level. The expression of PLC beta2 mRNA was not found in postnatal rat brains. The expression of PLC beta3 mRNA was found from P0, peaked at Pl4, and decreased to adult level in the purkinje cells of cerebellum. PLC beta4 mRNA was strongly expressed in the thalamus, cerebellum, cerebral cortex, and olfactory bulb. In these areas, the expression was gradually increased after birth, peaked at P2l, and decreased to adult level. In whole body parasagittal sections of 18 day old rat embryo, PLC betal mRNA was exclusively expressed in nervous tissue, PLC beta3 and PLC beta4 were expressed in various tissues, and the expression of PLC beta2 was not found in any kind of rat tissues. From the different spatiotemporal mRNA expression patterns of PLC beta isozymes in the postnatal rat brains, it is suspected that each PLC beta isozyme may have specific role in signal transduction for postnatal development of rat brain.
Adult ; Animals ; Brain* ; Cerebellum ; Cerebral Cortex ; Dentate Gyrus ; Embryonic Structures ; Hippocampus ; Humans ; Inositol ; Isoenzymes* ; Nervous System ; Olfactory Bulb ; Parturition ; Phospholipase C beta* ; Phospholipases* ; Purkinje Cells ; Putamen ; Rats* ; RNA, Messenger* ; Second Messenger Systems ; Signal Transduction ; Thalamus

During carcinogenesis, NF-kappaB mediates processes associated with deregulation of the normal control of proliferation, angiogenesis, and metastasis. Thus, suppression of NF-kappaB has been linked with chemoprevention of cancer. Accumulating findings reveal that heat shock protein 90 (HSP90) is a molecular chaperone and a component of the IkappaB kinase (IKK) complex that plays a central role in NF-kappaB activation. HSP90 also stabilizes key proteins involved in cell cycle control and apoptosis signaling. We have determined whether the exogenous administration of isoflavone-deprived soy peptide prevents 7,12-dimethylbenz[alpha]anthracene (DMBA)-induced rat mammary tumorigenesis and investigated the mechanism of action. Dietary administration of soy peptide (3.3 g/rat/day) significantly reduced the incidence of ductal carcinomas (50%), the number of tumors per multiple tumor-bearing rats (49%; P < 0.05), and extended the latency period of tumor development (8.07 +/- 0.92 weeks) compared to control diet animals (10.80 +/- 1.30; P < 0.05). Our results have further demonstrated that soy peptide (1) dramatically inhibits the expression of HSP90, thereby suppressing signaling pathway leading to NF-kappaB activation; (2) induces expression of p21, p53, and caspase-3 proteins; and (3) inhibits expression of VEGF. In agreement with our in vivo data, soy peptide treatment inhibited the growth of human breast MCF-7 tumor cells in a dose-dependent manner and induced apoptosis. Taken together, our in vivo and in vitro results suggest chemopreventive and tumor suppressive functions of isoflavone-deprived soy peptide by inducing growth arrest and apoptosis.
9,10-Dimethyl-1,2-benzanthracene ; Adenocarcinoma/*prevention & control ; Animals ; Apoptosis/*drug effects ; Breast Neoplasms/chemically induced/pathology/*prevention & control ; Cell Line, Tumor ; Chemoprevention ; Female ; Gene Expression Regulation, Neoplastic ; HSP90 Heat-Shock Proteins/genetics/metabolism ; Humans ; Isoflavones/chemistry ; NF-kappa B/genetics/metabolism ; Peptides/chemistry/isolation & purification/therapeutic use ; Rats ; Rats, Sprague-Dawley ; Soybean Proteins/chemistry/*isolation & purification/*therapeutic use ; Soybeans/chemistry

Mayaro virus (MAYV) is a mosquito-transmitted alphavirus that produces an acute, usually non-fatal, febrile illness including Mayaro fever. Like other alphaviruses, the MAYV E1 and E2 envelope glycoproteins are major viral surface antigens that play a key role in host recognition and infection. Here, we report expression and purification methods for recombinant MAYV E1 (rE1) and rE2 using a baculovirus system. Enzyme-linked immunosorbent assays (ELISA) revealed that rE1 and rE2 were antigenic and reacted with human anti-MAYV IgG and IgM. Cross-reactivity was also confirmed with human anti-Chikungunya virus (CHIKV) IgG and IgM. Furthermore, we developed an immunochromatographic strip test (IST) with rE2 to diagnose MAYV infection. Thus, purified rE2 may be valuable tool for rapidly diagnosing MAYV infection.

Ovarian cancer is a leading cause of death in women. Early detection of ovarian cancer is essential to decrease mortality. However, the early diagnosis of ovarian cancer is difficult due to a lack of clinical symptoms and suitable molecular diagnostic markers. Thus, identification of meaningful tumor biomarkers with potential clinical application is clearly needed. To search for a biomarker for the early detection of ovarian cancer, we identified human anterior gradient 2 (AGR2) from our systematic analysis of paired normal and ovarian tumor tissue cDNA microarray. We noted a marked overexpression of AGR2 mRNA and protein in early stage mucinous ovarian tumors compared to normal ovarian tissues and serous type ovarian tumors by Western blot analysis and immunohistochemistry. To further elucidate the role of AGR2 in ovarian tumorigenesis, stable 2774 human ovarian cancer cell lines overexpressing AGR2 were established. Forced expression of AGR2 in 2774 cells enhanced the growth and migration of ovarian cancer cells. AGR2 protein was detected in the serum of mucinous ovarian cancer patients by Western blot and ELISA analysis. Thus, AGR2 is a potential biomarker for the diagnosis of mucinous ovarian cancer and an ELISA assay may facilitate the early detection of mucinous ovarian cancer using patient serum.
Cell Line, Tumor ; Cell Movement/physiology ; Cell Proliferation ; Female ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; HEK293 Cells ; Humans ; Ovarian Neoplasms/genetics/*metabolism/pathology ; Proteins/genetics/*metabolism ; Tumor Markers, Biological/blood/*metabolism

OBJECTIVE: To prospectively investigate the feasibility of using 2 small intravenous catheters for high-rate computed tomography (CT) contrast injection in patients lacking superficial veins capable of accommodating ≤ 20-gauge catheters. MATERIALS AND METHODS: Sixty-eight consecutive eligible adults referred for dynamic liver CT were enrolled; 58 had previously undergone liver CT, including 8 that experienced extravasation. Two 22- or 24-gauge catheters were placed in all patients after 2–5 venipunctures, and 2 mL/kg of contrast agent (370 mg I/mL) was split-administered through both catheters to achieve total flow rate of 4 mL/s. Patients' experience and examination success rate, defined as uneventful scans completed at 4 mL/s or at < 4 mL/s achieving standard image quality in all phases, were analyzed. Quantitative hepatic signal-to-noise and hepatic vascular contrast-to-noise ratios (CNRs) were compared with 30 control examinations scanned at 4 mL/s using an 18-gauge catheter. RESULTS: One case each of extravasation and severe injection pain caused the examination to be aborted. Success rate was 88.2% (60/68; 54 patients scanned at 4 mL/s, 6 at 3.5–3.9 mL/s). Fifty-five of 58 patients (94.8%) that had past CT regarded the venipuncture as more tolerable than (n = 36) or similar to (n = 19) past experiences; 45 of 58 patients (77.6%) found contrast injection less painful than (n = 35) or similar to (n = 10) past experiences. When compared with control examinations, signal-to-noise ratio was similar in all phases (p ≥ 0.502), but the hepatic arterial CNR in arterial phase was slightly inferior (p ≤ 0.047). CONCLUSION: Using 2 small intravenous catheters can effectively achieve high-rate CT contrast injection in patients lacking adequate superficial veins.
Adult ; Catheters* ; Contrast Media ; Humans ; Liver ; Multidetector Computed Tomography ; Patient Satisfaction ; Phlebotomy ; Prospective Studies ; Quality Assurance, Health Care ; Signal-To-Noise Ratio ; Veins*