SummaryAfter approximately 50% of K from the 1–2 μm fraction of a naturally occurring microcrystalline muscovite was extracted with BaCl2 solution at 98°C electron-diffraction patterns showed a honeycomb-shaped diffuse scattering as well as intense diffuse streaks. The diffuse scattering became pronounced with increasing K-depletion. It may be interpreted in terms of a short-range order distribution of interlayer Ba ions, using an analogy to the work by Alcover et al. (1972, 1976). Since the diffuse streaks still remained after re-entry of K to the K-depleted specimen they might be related to structural disturbance, particularly in the basal oxygen arrangements, resulting from the K-extraction. Subsequent conversions of Ba-saturated vermiculitized muscovite into Na-, Mg-, and Ca-forms revealed distinct extra diffraction spots midway between two ordinary diffraction spots. They were broader than the ordinary spots and elongated along the three directions of [01]*, [11]*, and [11]*. The diffraction pattern containing the extra spots was, therefore, interpreted as a composite one made up by superimposing three independent patterns, each of which was composed of ordinary Bragg diffraction spots and extra diffraction maxima elongated in one of the three directions. This situation does not require a superlattice but modified the original face-centred lattice symmetry to a primitive lattice symmetry. However, the presence of a real superlattice should also be possible. The elongation and broadness of the extra diffraction maxima may be related to the orientation and size of coherent domains defined by a certain ordering of interlayer cationic distribution, which took place during the hydration processes including extraction and conversion.

The prognosis of HCC patients with extrahepatic metastases is poor. With regard to the cause of death, many patients would die of intrahepatic HCC and few of extrahepatic metastases. Although most of HCC patients with extrahepatic metastases should undergo treatment for the primary HCC mainly, treatment of extrahepatic metastases in selected HCC patients who have good hepatic reserve, intrahepatic tumor stage (T0-T2), and are free of portal venous invasion may improve survival.

Although high concentrations of reactive oxygen species (ROS) cause sperm pathology (ATP depletion leading to insufficient axonemal phosphorylation, lipid peroxidation and loss of motility and viability), recent evidence demonstrates that low and controlled concentrations of these ROS play an important role in sperm physiology. Reactive oxygen species, such as the superoxide anion, hydrogen peroxide and nitric oxide, induce sperm hyperactivation, capacitation or the acrosome reaction in vitro. The ROS involved in these processes may vary depending on experimental conditions, but all the evidence converges to describe these events as 'oxidative' or 'redox regulated'. Human sperm capacitation and acrosome reaction are associated with extracellular production of a superoxide anion that is thought to originate from a membrane 'oxidase'. The enzymes responsible for tyrosine phosphorylation-dephosphorylation of sperm proteins are possible targets for ROS since mild oxidative conditions cause increases in protein tyrosine phosphorylation and acrosome reaction. The lipid peroxidation resulting from low concentrations of ROS promotes binding to the zona pellucida and may trigger the release of unesterified fatty acids from the sperm plasma membrane. The fine balance between ROS production and scavenging, as well as the right timing and site for ROS production are of paramount importance for acquisition of fertilizing ability.

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The present study was designed to clarify the role of radical oxygen species in testicular germ cell apoptosis induced by heat stress. Testicular cells isolated from immature rats were cultured with or without elevated temperature, and occurrence of apoptosis in these cells was defined by the appearance of DNA fragmentation following agarose gel electrophoresis and by flow cytometric quantification of apoptotic cells. At 32.5 degrees C, < 1% of cells showed signs of apoptosis throughout the culture period, whereas under heat stress, the proportion of apoptotic cells increased to 5% at 37 degrees C after 24 h of culture, or to 14% after 1-h exposure at 43 degrees C followed by 23-h culture at 32.5 degrees C. Similar to the effect of heat stress, exogenously supplied oxygen free radicals also induced apoptosis. In contrast, treatment with catalase significantly attenuated heat stress-induced apoptosis. Furthermore, heat stress of testicular cells was associated with an increased intracellular peroxide level as measured by a fluorescent probe, 2', 7'-dichlorofluorescin diacetate. In conclusion, our data indicate the involvement of radical oxygen species during testicular germ cell apoptosis induced by heat stress. This study provides a useful in vitro model for the study of testicular germ cell apoptosis.

Although ghrelin acts as a modulator of feeding behavior and energy metabolism in the central nervous system, recent studies have implicated the peripheral actions of ghrelin in reproductive tissues. Here, we investigated the expression of ghrelin and its receptor (GHS-R) in mouse oocyte and preimplantation embryos, and we examined the role of ghrelin in the regulation of early embryo development. Both ghrelin and GHS-R mRNAs were detected in morula or more advanced embryo stages. As for the origin of ghrelin, both ghrelin mRNA and protein were identified in the uterine endometrium. The levels of ghrelin in uterine fluid as well as plasma were significantly increased in fasting mice compared with animals with free access to foods. Addition of ghrelin to culture media inhibited the development of two-cell embryos to the hatched blastocysts, and the inhibitory effects of ghrelin were abolished by an antagonist for the GHS-R. In addition, ghrelin significantly decreased the number of total cells, inner cell mass, and trophectoderm cells in blastocysts. These observations suggest that ghrelin could inhibit the development of preimplantation embryos during fasting. Thus, ghrelin may act as a peripheral factor to avoid the excess metabolic demands imposed by pregnancy during malnutritional states.

Leptin acts as a modulator of diverse reproductive functions, and recent studies have implicated involvement of leptin in the early embryo development in mammal. The aim of this study was to investigate the expression of leptin and its receptor (OB-R) in mouse oocyte and preimplantation embryo, and to examine whether leptin influenced the early embryo development. Leptin mRNA was detected in blastocyst and hatched blastocyst, and two splice variants of OB-R (OB-Ra and OB-Rb) mRNAs were detected in oocytes, 1-cell, 2-cell, morula, blastocyst, and hatched blastocyst. As for the origin of leptin, both leptin mRNA and protein were identified in the oviduct epithelium and endometrium of pregnant mouse. In the pregnant mouse, the levels of leptin in uterine fluid were higher than those in nonpregnant mouse. Addition of leptin to embryo culture media promotes the development from 2-cell stage embryos to the blastocysts, fully expanded blastocysts and hatched blastocysts. This effect was neutralized by an antibody against the extracellular domain of OB-R. Leptin significantly increased the total cell number of blastocysts, and the effect was preferentially observed in the trophectoderm. These findings raise the possibility of a paracrine/autocrine leptin signaling system regulating the development of mouse preimplantation embryo.

B-RTO is useful for treatment of bleeding gastric varices, achieving high eradication of gastric varices, a low rebleeding rate, and a fairly good prognosis with improved hepatic function.