Sex differences in interactions between nucleus accumbens and visual cortex by explicit visual erotic stimuli: an fMRI study.

Men tend to have greater positive responses than women to explicit visual erotic stimuli (EVES). However, it remains unclear, which brain network makes men more sensitive to EVES and which factors contribute to the brain network activity. In this study, we aimed to assess the effect of sex difference on brain connectivity patterns by EVES. We also investigated the association of testosterone with brain connection that showed the effects of sex difference. During functional magnetic resonance imaging scans, 14 males and 14 females were asked to see alternating blocks of pictures that were either erotic or non-erotic. Psychophysiological interaction analysis was performed to investigate the functional connectivity of the nucleus accumbens (NA) as it related to EVES. Men showed significantly greater EVES-specific functional connection between the right NA and the right lateral occipital cortex (LOC). In addition, the right NA and the right LOC network activity was positively correlated with the plasma testosterone level in men. Our results suggest that the reason men are sensitive to EVES is the increased interaction in the visual reward networks, which is modulated by their plasma testosterone level.

Epigenetic characteristics of bovine donor cells for nuclear transfer: levels of histone acetylation.

Donor cell type, cell-cycle stage, and passage number of cultured cells all affect the developmental potential of cloned embryos. Because acetylation of the histones on nuclear chromatin is an important aspect of gene activation, the present study investigated the differences in histone acetylation of bovine fibroblast and cumulus cells at various passages and cell-cycle stages. The acetylation was qualitatively analyzed by epifluorescent confocal microscopy and quantitatively by immunofluorescent flow cytometry. Specifically, we studied levels of histone H4 acetylated at lysine 8 and histone H3 acetylated at lysine 18; acetylation at these lysine residues is among the most common for these histone molecules. We also studied levels of linker histone H1 in donor cells. Our results show that stage of cell cycle, cell type, and number of cell passages all had an effect on histone content. Histone H1 and acetyl histone H3 increased with cell passage (passages 5-15) in G0/G1- and G2/M-stage cumulus and fibroblast cells. We also found that acetyl histone H4 was lower in early versus late cell passages (passage 5 vs. 15) for G0/G1-stage cumulus cells. In both cell types examined, acetyl histones increased with cell-cycle progression from G0/G1 into the S and G2/M phases. These results indicate that histone acetylation status is remodeled by in vitro cell culture, and this may have implications for nuclear transfer.

Cysteine, glutathione, and Percoll treatments improve porcine oocyte maturation and fertilization in vitro.

Several factors have been examined to improve in vitro fertilization and development of porcine oocytes. Cysteine is known to be beneficial for oocyte maturation and male pronuclear formation in pigs and glutathione is known to help prevent membrane disruption of sperm in other species, including human. It has also been reported that the presence of cumulus cells influences the outcome of in vitro fertilization in cattle. The objectives of the present study were to investigate the effects of several factors involved in porcine in vitro maturation (IVM) and fertilization (IVF) procedures on oocyte embryogenic competence. The following factors were examined: the effects of different concentrations (0, 0.285, 0.57, 1.14, 2.28 microM) and exposure duration (22 and 44 hr) of cysteine during IVM, glutathione inclusion and of cumulus presence during IVF, and the use of gradient Percoll (45%/90%) during sperm preparation. The presence of cysteine in maturation medium improved blastocyst development significantly regardless of the duration of exposure when compared to the control (11--16% vs. 4%, P < 0.01). However, no dose-responsive effect was observed at the concentrations tested. The use of gradient Percoll during sperm preparation significantly improved cleavage (85% vs. 57%, P < 0.01) and blastocyst development (24% vs. 6%, P < 0.01) over conventional sperm preparation. Significant improvement was also achieved by the addition of glutathione to Percoll gradient (30% vs. 20%, P < 0.05). In conclusion, cysteine and glutathione as well as Percoll and cumulus were beneficial to embryogenic competence of porcine oocytes in this study. Mol. Reprod. Dev. 59:330-335, 2001.

Dynamic changes of cumulus-oocyte cell communication during in vitro maturation of porcine oocytes.

Oocyte maturation is a key issue of current animal biotechnology. This study was designed to examine the morphodynamics of the cumulus-oocyte association during oocyte maturation. Porcine cumulus-oocyte complexes were recovered from slaughterhouse ovaries; matured in vitro for 0, 24, 36, and 44 h; and evaluated by scanning electron microscopy either combined or not combined with the osmium-dimethyl sulfoxide-osmium maceration (ODO) method. The cytoskeleton distribution was also observed by fluorescence staining. Prior to maturation culture (0 h), the spherical cumulus cells were tightly clustered around the oocyte, with narrow intercellular spaces. They showed active secretion at 36 h and were fully expanded at 44 h of culture. The ODO methods revealed that the cumulus cells projected numerous long and thin transzonal projections at 0 h, but these were largely disconnected at 44 h. The outer surface of the zona pellucida showed a meshwork surface regardless of time of incubation, whereas the inner surface changed from a fine fibrous surface to a spongy surface that was coated with mucin. The vitelline surface changed from a sparse distribution of short microvilli (MV) to a dense distribution of well-developed MV. Fluorescence staining showed that the cumulus cell projections consisted mainly of microfilaments, which were abundant at the germinal vesicle and metaphase-I (M-I) stages (0-24 h) but which were decreased in number at the M-II stage (36-44 h). We conclude that the cumulus-oocyte transzonal projections became disconnected between the M-I and M-II stages as a result of cumulus expansion. The cumulus-cumulus communications, however, remained intact at these stages, although the biological functions of these communications were not clear.

Antimutagenic property of an herbal medicine, Polygonum multiftorum Thunb. detected by the Tradescantia micronucleus assay.

The root extracts of a Chinese herb, Polygonum multiflorum Thunb., have been used for centuries as an internal medicine to improve liver and kidney functions. In this study, we evaluated the antimutagenic property of this drug with the Tradescantia micronucleus (Trad-MCN) assay. The Trad-MCN bioassay is a well-established test for chromosome damage induced by physical or chemical agents in terms of micronuclei (MCN) frequency. Inflorescences of the Tradescantia plant cuttings were first exposed to 0.35 Gy soft X-rays (80 kV, 5 mA, 1 mm Al filter, dose rate around 0.50 Gy/min), followed by drug treatments at 1, 3, and 6% concentrations of the aqueous solution for a total recovery period of 24 hours. The positive (X-rays), negative (nutrient solution), and drug control (3% drug solution) groups were maintained in each of the three series of repeated experiments. Flower buds of the treated and control groups were fixed in aceto-alcohol (1:3 ratio) in preparation for slides to score MCN frequencies in the early tetrads of the meiotic pollen mother cells. The mean MCN frequencies (MCN/100 tetrads +/- SE) of the positive control (irradiated) was 26.68 +/- 2.49; the negative control was 2.93 +/- 0.50; the PM solution control was 2.06 +/- 0.39, and the 0.35 Gy X-ray plus 6% PM drug treated was 18.76 +/- 1.69. A 45% reduction in chromosome damage was observed. Antimutagenic effects were relatively decreased at lower concentrations of PM. This antimutagenic effect could be attributed to the antioxidant action of PM, enhancement of DNA repair, or the radical elimination from the irradiated plant cells.

In situ monitoring with the Tradescantia bioassays on the genotoxicity of gaseous emissions from a closed landfill site and an incinerator.

A dual monitoring system composed of the Tradescantia-Micronucleus (Trad-MCN) and Tradescantia-Stamen-Hair-Mutation (Trad-SHM) bioassays was utilized to monitor directly the genotoxicity of the gaseous emission at a closed landfill site and around an incinerator. Four of the commonly emitted gaseous agents from the landfill flare pipes, i.e. toluene, ethylbenzene, trichloroethylene and ethyltoluene were also evaluated for their genotoxicity in the laboratory. The in situ monitoring trips (360 km one way) were carried out by transporting the plant cuttings in a clean air box or in an air-tight plastic bag to the site and exposing these test cuttings for 5-7 h. The exposed plant samples were examined for micronuclei frequencies or the pink mutation rate after the appropriate recovery periods (24 h for MCN, 7-11 days for SHM). A total of 20 monitoring trips were made to the landfill, and 8 to the nearby surroundings (100-500 m from the chimney) of the incinerator site in a two year period. The major findings of the Trad-MCN test on the clastogenicity of the gaseous emission from the flare pipe of the landfill site showed positive responses or toxic effects in 6 out of 20 trips, and that from the incinerator showed positive responses in 5 out of the 8 trips. These positive responses were closely associated with the weather, i.e. low wind velocity, high temperature and relative humidity, and especially the distance from the chimney of the incinerator. The MCN frequencies and mutation rates of the Elementary School site (E. Sch) which is about 200 m from the fence of the landfill site were mostly negative, except the test results of three trips. Trad-SHM tests on the mutagenicity of gaseous emissions from the flare pipe of the landfill showed 12 positive responses out of 20 trials and 2 positives out of 4 trials from the incinerator gaseous emissions. The average mutation rate from 20 Trad-SHM monitoring trips is positive when the ANOVA and Dunnett's t-statistic were applied to the consolidated data. There is a significant (0.01) difference between the lab control and the gas exposed groups, and between the field control and gas exposed groups. Results of the Trad-SHM test at the E. Sch. site were mostly negative except for one trip. In general, micronuclei frequencies and mutation rates of the field control groups were relatively higher than those of the lab controls. The Trad-MCN test on pure gases showed positive responses in all 3 repeated tests on toluene (50-892 ppm). The test results of ethylbenzene yielded positive responses at 172 ppm/min and 1549 ppm/min dosages and exhibited toxicity at higher concentrations. Trad-MCN tests on trichloroethylene and ethyltoluene yielded positive responses at around 100-200 ppm/min level. Three repeated Trad-SHM tests on toluene yielded no positive response at low concentrations (4.3-12.9 ppm).