Characterization of 1-methyladenine binding in starfish oocyte cortices

1-Methyladenine (1MeAde) is the naturally occurring maturation-inducing hormone of starfish oocytes. We have prepared a biologically active [3H]1MeAde of high purity and relatively high specific radioactivity. This ligand binds to cortices isolated from full-grown prophase-arrested oocytes of the starfish Asterina pectinifera. The binding of [3H]1MeAde to cortices was rapid and reached equilibrium in approximately 15 min. This is in excellent agreement with the hormone-dependent period required for the induction of oocyte maturation. Binding was maximal between pH 6.4 and 8.0 and diminished sharply above and below this range. Analysis of Scatchard plots of the equilibrium binding of [3H]1MeAde to cortices indicates the existence of a single class of binding site with a dissociation constant of 0.3 microM and a binding capacity of 0.02 fmol per cortex. Whereas biologically active analogs (1-benzyladenine, 1-ethyladenine) inhibited the specific binding of [3H]1MeAde by cortices, biologically inactive analogs (2-methyladenine, 3-methyladenine, 1,9-dimethyladenine, and 1-methylhypoxanthine) did not. These results suggest that the 1MeAde binding characterized herein is necessary for the maturational action of 1MeAde on starfish oocytes.     

Involvement of gonadotrophin and steroid hormones in spermiation in the amago salmon, Oncorhynchus rhodurus, and goldfish, Carassius auratus

Effects of intraperitoneal injections of chum salmon gonadotrophin (SGA) and various steroid hormones (17α,20β-dihydroxy-4-pregnen-3-one; 17α,20β-diOHprog, 17α-hydroxy-progesterone, testosterone, 11-ketotestosterone) on the induction of in vivo spermiation were examined in nonspermiating amago salmon (Oncorhynchus rhodurus) and goldfish (Carassius auratus). A single injection of SGA to amago salmon and two successive injections to goldfish induced strong spermiation responses. Similarly two successive injections of 17α, 20β-diOHprog caused precocious spermiation in both species; however, the response to 17α,20β-diOHprog was of lesser magnitude than that to SGA. The spermiation response of goldfish to 17α-hydroxyprogesterone was similar to that of 17α,20β-diOHprog. Neither testosterone nor 11-ketotestosterone were effective in inducing spermiation in amago salmon, but these steroids were found to be slightly effective in goldfish. Effects of a single injection of SGA on changes in serum levels of 17α,20β-diOHprog and 11-ketotestosterone were also examined in nonspermiating amago salmon. Serum levels of 17α,20β-diOHprog dramatically increased after treatment and peaked on Day 2 and thereafter declined quickly. Similarly, 11-ketotestosterone peaked on Day 2, but the levels remained high throughout the experimental period. Considered together, these findings are consistent with the suggestion that 17α,20β-diOHprog is involved in the process of spermiation in teleosts. It is further suggested that this hormone is a testicular steroidal mediator of gonadotrophin-induced spermiation in amago salmon.     

A case of sarcoidosis exacerbated after acute hepatitis C]

We describe a case of sarcoidosis exacerbated after acute hepatitis C, with particular reference to past case reports. A 30-year-old woman was admitted to our hospital in June 1997 because of cough and chest discomfort. Sarcoidosis was diagnosed as a result of chest radiography findings, transbronchial lung biopsy and bronchoalveolar lavage. A month later she was re-admitted because of fatigue. Acute hepatitis C was diagnosed from the findings of liver histology and HCV RNA. On the 42 nd day of hospitalization she complained of a severe cough, and a chest radiograph showed aggravation of bilateral reticulonodular shadows. This case suggested that acute hepatitis C may lead to an exacerbation of sarcoidosis. On August 4, 1999, the bilateral reticulonodular shadows had disappeared from the chest radiographs. Acute hepatitis C became chronic, but subsequently resolved spontaneously.     

Adenovirus-mediated gene transfer of triple human complement regulating proteins (DAF, MCP and CD59) in the xenogeneic porcine-to-human transplantation model

In this study, the adenovirus-mediated gene transfer of triple human complement regulating proteins was investigated in xenogeneic pig liver perfusion. The porcine liver was perfused in situ at 4 degrees C under a pump-driven veno-venous shunt of the portal vein and inferior vena cava, with 5 to 15x10(11) plaque-forming units (pfu) of adenovirus vector (group 1: AxCALacZ; 2: AxCACD59; 3: AxCACD59 + AxCADAF; 4: AxCACD59 + AxCADAF + AxCAMCP) for 1 h (for each, n=3). The livers were harvested 24 h after gene transfer and then were reperfused ex-vivo with fresh human blood for 2 h. In immunohistochemical staining, each complement regulating protein (CRP) showed a distribution similar to that of the LacZ expression. The C3 levels in the perfusate were also maintained at higher levels in group 4 from 60 to 120 min after reperfusion (C3: 85% to 95% of the initial level) than in groups 1 to 3 (C3: 80% to 90% of the initial level) from 60 to 120 min after reperfusion. The complement deposition on the porcine liver [C3, membrane attack component (MAC)] decreased significantly more in group 4 than in groups 1 to 3. In conclusion, the adenovirus-mediated multiple gene transfer of human CRPs (hCRPs) was found to effectively suppress the complement activation in xenogeneic pig liver perfusion.     

Visibility of gallstone fragments at US and fluoroscopy: implications for monitoring gallstone lithotripsy

To assess the value of ultrasound (US), fluoroscopy, and spot radiography in the detection, counting, and measurement of gallstone fragments during lithotripsy, in vitro visibility studies were conducted on fragments from 20 stones. Fluoroscopic visibility was evaluated during and after lithotripsy on 185 fragments placed in an anthropomorphic phantom. Three US experiments were performed on the fragments to study the visibility of fragments as a function of size, the accuracy of the count with large numbers of fragments, and the ability of observers to detect and count fragments larger than both 4 mm and 5 mm. With fluoroscopy, fragment detection rates ranged from 20% (fragments larger than 2.5 mm) to 80% (fragments larger than 4.5 mm). With US, all fragments larger than 1.5 mm were detected, and US was significantly better than fluoroscopy and spot radiography for detection of fragments 2.5 mm or smaller. US was also more accurate than fluoroscopy (11% vs 59% error) in the assessment of the number of fragments. When fragments larger than 4 mm or 5 mm were being counted with US, 92% of the fragments were visualized. The results suggest that US is more accurate for monitoring gallstone lithotripsy than fluoroscopy or spot radiography.     

Susceptibility of brain cells to murine cytomegalovirus infection in the developing mouse brain

summary Mouse embryos were infected with murine cytomegalovirus (MCMV) by injecting the virus into the cerebral ventricles in the late stage of gestation; the brains of the offspring were then analyzed using the histological and immunohistochemical methods. Brains of the offspring, which were injected with relatively high titers of MCMV [1×10⁴ plaque-forming units (pfu)] on day 13 of gestation exo utero or on day 15 of gestation in utero, showed massiv necrosis of the cerebral cortex with gliomesodermal proliferation around 9 to 10 days after birth. In these brains, viral antigen-positive cells were observed in zonal arrangement in the lesion-free cortex and in the hippocampus. Immunohistochemical double staining showed that some of the viral antigen-positive cells had also reacted with antibody to neuron-specific enolase at the same time, but had hardly reacted with antibodies to braintype creatine kinase or glial fibrillary acidic protein. Brains of the offspring, which were injected with relatively low titers of virus (1×10³ pfu) on day 15 of gestation, showed zonal arrangement of viral antigenpositive cells mainly in the cerebral cortex and in the hippocampus 7 days after birth, although the numbers of the positive cells were low. Fourteen days after birth, some of these offspring showed atrophy of the cerebral cortex and the hippocampus. These results suggest that some of the neuronal cells in the cerebral cortex and the hippocampus have special susceptibility to MCMV infection.Supported in part by a Grant (87-05) from the National Center of Neurology and Psychiatry (NCNP) of the Ministry of Health and Welfare of Japan