Stem Cell-Like Differentiation Potentials of Endometrial Side Population Cells as Revealed by a Newly Developed In Vivo Endometrial Stem Cell Assay

Background

Endometrial stem/progenitor cells contribute to the cyclical regeneration of human endometrium throughout a woman''s reproductive life. Although the candidate cell populations have been extensively studied, no consensus exists regarding which endometrial population represents the stem/progenitor cell fraction in terms of in vivo stem cell activity. We have previously reported that human endometrial side population cells (ESP), but not endometrial main population cells (EMP), exhibit stem cell-like properties, including in vivo reconstitution of endometrium-like tissues when xenotransplanted into immunodeficient mice. The reconstitution efficiency, however, was low presumably because ESP cells alone could not provide a sufficient microenvironment (niche) to support their stem cell activity. The objective of this study was to establish a novel in vivo endometrial stem cell assay employing cell tracking and tissue reconstitution systems and to examine the stem cell properties of ESP through use of this assay.

Methodology/Principal Findings

ESP and EMP cells isolated from whole endometrial cells were infected with lentivirus to express tandem Tomato (TdTom), a red fluorescent protein. They were mixed with unlabeled whole endometrial cells and then transplanted under the kidney capsule of ovariectomized immunodeficient mice. These mice were treated with estradiol and progesterone for eight weeks and nephrectomized. All of the grafts reconstituted endometrium-like tissues under the kidney capsules. Immunofluorescence revealed that TdTom-positive cells were significantly more abundant in the glandular, stromal, and endothelial cells of the reconstituted endometrium in mice transplanted with TdTom-labeled ESP cells than those with TdTom-labeled EMP cells.

Conclusions/Significance

We have established a novel in vivo endometrial stem cell assay in which multi-potential differentiation can be identified through cell tracking during in vivo endometrial tissue reconstitution. Using this assay, we demonstrated that ESP cells differentiated into multiple endometrial lineages in the niche provided by whole endometrial cells, indicating that ESP cells are genuine endometrial stem/progenitor cells.     

A U-Box E3 Ubiquitin Ligase,PUB20, Interacts with the Arabidopsis G-Protein β Subunit,AGB1

An Arabidopsis U-box E3 ubiquitin ligase Plant U-box 20 (PUB20; alternatively called AtCMPG1) was identified as a possible interactor of the Arabidopsis G-protein β subunit, AGB1, by yeast two-hybrid screening. A bimolecular fluorescence complementation (BiFC) assay showed that PUB20 interacted with AGB1 in the nuclei and the cytosol. The expression levels of PUB20 and its closest homolog, PUB21 were stable under many conditions. GUS driven by the PUB20 promoter was active in anthers, pollen, premature seeds and receptacles and GUS driven by the PUB21 promoter was active in anthers and funiculi. PUB20 was found to have autoubiquitination activity in vitro.     

Physiological functions of the effects of the different bathing method on recovery from local muscle fatigue

Background

Recently, mist saunas have been used in the home as a new bathing style in Japan. However, there are still few reports on the effects of bathing methods on recovery from muscle fatigue. Furthermore, the effect of mist sauna bathing on human physiological function has not yet been revealed. Therefore, we measured the physiological effects of bathing methods including the mist sauna on recovery from muscle fatigue.

Methods

The bathing methods studied included four conditions: full immersion bath, shower, mist sauna, and no bathing as a control. Ten men participated in this study. The participants completed four consecutive sessions: a 30-min rest period, a 10-min all out elbow flexion task period, a 10-min bathing period, and a 10-min recovery period. We evaluated the mean power frequency (MNF) of the electromyogram (EMG), rectal temperature (Tre), skin temperature (Tsk), skin blood flow (SBF), concentration of oxygenated hemoglobin (O2Hb), and subjective evaluation.

Results

We found that the MNF under the full immersion bath condition was significantly higher than those under the other conditions. Furthermore, Tre, SBF, and O2Hb under the full immersion bath condition were significantly higher than under the other conditions.

Conclusions

Following the results for the full immersion bath condition, the SBF and O2Hb of the mist sauna condition were significantly higher than those for the shower and no bathing conditions. These results suggest that full immersion bath and mist sauna are effective in facilitating recovery from muscle fatigue.     

Physiological responses at short distances from a parametric speaker

In recent years, parametric speakers have been used in various circumstances. In our previous studies, we verified that the physiological burden of the sound of parametric speaker set at 2.6 m from the subjects was lower than that of the general speaker. However, nothing has yet been demonstrated about the effects of the sound of a parametric speaker at the shorter distance between parametric speakers the human body. Therefore, we studied this effect on physiological functions and task performance. Nine male subjects participated in this study. They completed three consecutive sessions: a 20-minute quiet period as a baseline, a 30-minute mental task period with general speakers or parametric speakers, and a 20-minute recovery period. We measured electrocardiogram (ECG) photoplethysmogram (PTG), electroencephalogram (EEG), systolic and diastolic blood pressure. Four experiments, one with a speaker condition (general speaker and parametric speaker), the other with a distance condition (0.3 m and 1.0 m), were conducted respectively at the same time of day on separate days. To examine the effects of the speaker and distance, three-way repeated measures ANOVA (speaker factor x distance factor x time factor) were conducted. In conclusion, we found that the physiological responses were not significantly different between the speaker condition and the distance condition. Meanwhile, it was shown that the physiological burdens increased with progress in time independently of speaker condition and distance condition. In summary, the effects of the parametric speaker at the 2.6 m distance were not obtained at the distance of 1 m or less.     

Effects of blue pulsed light on human physiological functions and subjective evaluation

Background

It has been assumed that light with a higher irradiance of pulsed blue light has a much greater influence than that of light with a lower irradiance of steady blue light, although they have the same multiplication value of irradiance and duration. We examined the non-visual physiological effects of blue pulsed light, and determined whether it is sensed visually as being blue.

Findings

Seven young male volunteers participated in the study. We placed a circular screen (diameter 500 mm) in front of the participants and irradiated it using blue and/or white light-emitting diodes (LEDs), and we used halogen lamps as a standard illuminant. We applied three steady light conditions of white LED (F0), blue LED + white LED (F10), and blue LED (F100), and a blue pulsed light condition of a 100-μs pulse width with a 10% duty ratio (P10). The irradiance of all four conditions at the participant''s eye level was almost the same, at around 12 μW/cm². We measured their pupil diameter, recorded electroencephalogram readings and Kwansei Gakuin Sleepiness Scale score, and collected subjective evaluations. The subjective bluish score under the F100 condition was significantly higher than those under other conditions. Even under the P10 condition with a 10% duty ratio of blue pulsed light and the F10 condition, the participant did not perceive the light as bluish. Pupillary light response under the P10 pulsed light condition was significantly greater than under the F10 condition, even though the two conditions had equal blue light components.

Conclusions

The pupil constricted under the blue pulsed light condition, indicating a non-visual effect of the lighting, even though the participants did not perceive the light as bluish.     

Can breakfast tryptophan and vitamin B6 intake and morning exposure to sunlight promote morning-typology in young children aged 2 to 6 years?

ABSTRACT: This study tried to examine, from epidemiological and physiologic anthropological (Japanese culture on breakfast) points of view, the integrated effects of the amount of tryptophan and vitamin B6 intake and the following exposure to sunlight on the circadian typology and sleep habits in young Japanese children aged 2 to 6 years, using the newly-evaluated calculating system of tryptophan (Tryptophan Index 2009) and vitamin B6 intake (VitaminB6 Index 2009) at breakfast. The positive and significant correlation was shown between the Morningness-Eveningness (M-E) score and the Tryptophan Index and also the Vitamin B6 Index. This positive correlation between M-E score and amount of tryptophan intake was shown only by children who were exposed to sunlight for longer than 10min after breakfast. These results might support the following hypothesis: higher tryptophan and vitamin B6 intake at breakfast could promote the synthesis of serotonin via light stimulation in the morning in children.     

Evaluation of a synthetic C34 trimer of HIV-1 gp41 as AIDS vaccines

An artificial antigen forming the C34 trimeric structure targeting membrane-fusion mechanism of HIV-1 has been evaluated as an HIV vaccine. The C34 trimeric molecule was previously designed and synthesized using a novel template with C3-symmetric linkers by us. The antiserum produced by immunization of the C34 trimeric form antigen showed 23-fold higher binding affinity for the C34 trimer than for the C34 monomer and showed significant neutralizing activity. The present results suggest effective strategies of the design of HIV vaccines and anti-HIV agents based on the native structure mimic of proteins targeting dynamic supramolecular mechanisms in HIV fusion.     

Synthetic studies of bi-fluorescence-labeled maltooligosaccharides as substrates for α-amylase on the basis of fluorescence resonance energy transfer (FRET)

A series of bi-fluorescence-labeled maltooligosaccharides that lead to fluorescence resonance energy transfer (FRET) was systematically synthesized. Effective FRETs were observed with all of the synthesized probes. Digestion of probes having tetra-, quintet-, hexa- or hepta-saccharidic chain lengths with human saliva α-amylase resulted in disappearance of FRET when an excitation wavelength of at 290nm was used followed by detection at ca. 520nm due to emission from the dansyl moiety. However, continuous FRET was observed when probes having di- or trisaccharidic chain lengths were used as substrates. In addition to the substrate characteristics based on saccharidic chain length, the reaction rates of digestion for the substrates by amylase were different and also depended on their saccharidic chain length.