Diversity of genome size and chromosome number in homothallic and heterothallic strains of the Closterium peracerosum-strigosum-littorale complex (Desmidiales, Zygnematophyceae, Streptophyta).

The evolutionary transitions of mating systems between outcrossing and self-fertilization are often suggested to associate with the cytological and genomic changes, but the empirical reports are limited in multicellular organisms. Here we used the unicellular zygnematophycean algae, the Closterium peracerosum-strigosum-littorale (C. psl.) complex, to address whether genomic properties such as genome sizes and chromosome numbers are associated with mating system transitions between homothallism (self-fertility) and heterothallism (self-sterility). Phylogenetic analysis revealed the polyphyly of homothallic strains, suggesting multiple transitions between homothallism and heterothallism in the C. psl. complex. Flow cytometry analysis identified a more than 2-fold genome size variation, ranging from 0.53 to 1.42 Gbp, which was positively correlated with chromosome number variation between strains. Although we did not find consistent trends in genome size change and mating system transitions, the mean chromosome sizes tend to be smaller in homothallic strains than in their relative heterothallic strains. This result suggests that homothallic strains possibly have more fragmented chromosomes, which is consistent with the argument that self-fertilizing populations may tolerate more chromosomal rearrangements.

Two cases of bilateral reverse shoulder arthroplasty performed in patients with rheumatoid arthritis.

Bilateral shoulder joint disorders caused by rheumatoid arthritis significantly impair daily functioning owing to a lack of contralateral compensation. In Japan, reverse shoulder joint prostheses were approved in 2014. This was expected to improve the surgical outcomes of rheumatoid shoulder arthroplasty. We report two patients with rheumatoid arthritis who underwent bilateral reverse shoulder arthroplasty. This study aims to evaluate their postoperative clinical outcomes and activities of daily living. The patients were women in their 70s with stage III class 2 rheumatoid arthritis. Their treatment and postoperative activities of daily living were retrospectively reviewed. The first patient underwent the inlay type and experienced a residual limitation of external rotation postoperatively; therefore, she was restricted to dress with front-open clothes. However, she was able to undress after the lining of the garment was changed to a slippery material. The second patient underwent the onlay type and showed almost no limitations in postoperative activities of daily living. She was able to undress with an external rotation of 40-50°. Bilateral reverse shoulder arthroplasty improved range of motion, the Japanese Orthopaedic Association shoulder score, and functional outcomes. Only a few difficulties were encountered in the activities of daily living.

Methoxyflavones isolated from the whole plant of Scutellaria rubropunctata Hayata var. rubropunctata promote osteoblast differentiation in MC3T3-E1 cells.

In this study, we isolated two new methoxyflavones (1 and 2) and eight known methoxyflavones (3-10) from the whole plant of Scutellaria rubropunctata Hayata var. rubropunctata (SR). Based on spectroscopic analyses, the methoxyflavones were identified as 5,8,2',6'-tetramethoxy-6,7-methylenedioxyflavone (1) and 5,2',6'-trimethoxy-6,7-methylenedioxyflavone (2). We reported SR might have effects on promoting osteoblast differentiation and stimulating estrogen receptor (ER) in the previous study. Then, the effects of 1-10 on pre-osteoblast MC3T3-E1 cells were investigated, and 1, 2, and 9 were observed to promote alkaline phosphatase activity. To evaluate their effect on osteogenesis-related genes, we performed gene expression analysis using quantitative real-time PCR after treatment of MC3T3-E1 cells with these compounds. Although 2 was only effective at lower concentrations, 1 and 9 upregulated the mRNA levels of Runx2, Osterix, Osteopontin, Osteocalcin, Smad1, and Smad4. These results indicate that 1 and 9 may induce osteoblast differentiation by activating Runx2 via the BMP/Smad pathway and may play a central role in the promotion of osteoblast differentiation by SR. The ER agonist activity of 1-10 were tested using a luciferase reporter assay in HEK293 cells. However, none of the compounds exhibited remarkable activity. Thus, SR may contain other compounds that contribute to its ER agonist activity.

Effects of a Whole Plant Extract of Scutellaria rubropunctata var. rubropunctata on Bone Metabolism with Estrogen Receptor Activation.

We screened natural resources for estrogen receptor (ER)-activating and bone metabolism-promoting activities with the aim of finding potential treatments for osteoporosis. A screen of 1531 extracts from Ryukyu Arc plants using a luciferase reporter assay identified an 80% MeOH extract of Scutellaria rubropunctata var. rubropunctata (SRE) with dose-dependent ER transcription-promoting activity. Importantly, SRE had no proliferative effect on human breast cancer cells. SRE enhanced the ALP activity of pre-osteoblast MC3T3-E1 cells after 72 h in culture and slightly enhanced mineralization at 14 and 21 d. SRE did not significantly affect the TRAP activity of RAW264.7 cells. Gene expression analysis in MC3T3-E1 cells by quantitative real-time PCR revealed that SRE upregulated the mRNA levels of Runx2, Osterix (Osx), Osteopontin (Opn), Osteocalcin (Ocn), Smad1, Smad4, and Smad5 at 72 h, and those of Runx2, Osx, Smad1, Smad4, and Smad5 at 21 d of osteogenic induction. Analysis of the expression levels of osteogenic markers suggested that SRE may promote osteogenic differentiation by acting at the early stage of differentiation rather than at the late stage of differentiation. These results indicate that SRE activates ER and induces osteoblast differentiation by activating Runx2 and Osx through the BMP/Smad pathway, suggesting that SRE may be useful for the prevention and treatment of postmenopausal osteoporosis.

[Adsorption of Airbone Organophosphorus Flame Retardants on Polished Rice Stored in a House].

In order to clarify the actual situation of indoor pollution to polished rice with organophosphorus flame retardants (PFRs) which are ubiquitous in the indoor environment, the pollution of PFRs to polished rice stored in a house for a week was investigated. The survey covered 64 ordinary families in the Osaka region. We analyzed six typical PFRs in 37 rice samples in 2015, and ten typical PFRs in 27 rice samples in 2016. Polished rice was homogenized with acetone-hexane and defatted by hexane-acetonitrile partition. Quantitative analysis for PFRs was performed by GC-FPD. The detection frequency of PFRs in the rice samples was 35/37 in 2015 and 27/27 in 2016. The highest values were 160 ng/g for TCEP, 500 ng/g for TCIPP and 430 ng/g for TBEP. The concentration ratio of each detected PFR in the polished rice samples was different in each house. In the analysis of 16 polished rice samples which were stored in the home, PFRs were detected in ten samples regardless of the storage methods. PFRs were detected from 12 out of 16 commercial brown rice samples. This result suggested that they were polluted during distribution and storage process of them.

[Chlorpyrifos-Methyl, Pirimiphos-Methyl and Fenitrothion Residues in Commercial Wheat Products].

We measured the concentrations of organophosphorus pesticide （chlorpyrifos-methyl, pirimiphos-methyl and fenitrothion） residues in domestic and imported commercial wheat products （flour, noodles, cookies） collected from 2008 to 2016. Chlorpyrifos-methyl and fenitrothion in domestic flour samples were detected in 16 out of 34 samples at levels of ＜0.001 to 0.016 ppm, and in 14 out of 34 samples at levels of ＜0.001 to 0.004 ppm. Chlorpyrifos-methyl was detected in 22 out of 38 domestic cookies at levels of ＜0.001 to 0.054 ppm （median: 0.001 ppm）. Relatively high concentrations of chlorpyrifos-methyl （0.005 to 0.054 ppm） were found in six domestic cookies containing wheat bran. Pirimiphos-methyl was detected in 32 out of 68 cookies from foreign countries at levels of ＜0.001 to 0.11 ppm. Pirimiphos-methyl was detected frequently in products imported from Europe.

[Determination of S-421 in Commercial Fish and Shellfish].

S-421 is a synergist of pyrethroid and organophosphorus pesticides, which are used as termiticides or household insecticides. S-421 is stable and ubiquitous in the environment. Here we describe the concentrations of S-421 in domestic and imported commercial fish collected from 2009 to 2016. Samples were extracted with acetone/hexane and S-421 was purified on a silica gel column. Quantitative analysis was performed by GC-ECD. S-421 was detected in 78 of 116 samples of domestic fish and shellfish at levels of <0.2 to 2.6 ng/g (mean: 0.4 ng/g), and in 69 of 102 imported samples at a level of <0.2 to 1.5 ng/g (mean: 0.4 ng/g). The concentrations of S-421 in fish were lower than those of p,p'-DDE and similar to those of ß-HCH.

Identification and characterization of natural microbial products that alter the free d-aspartate content of mammalian cells.

Mammalian cells possess the molecular apparatus necessary to take up, degrade, synthesize, and release free d-aspartate, which plays an important role in physiological functions within the body. Here, biologically active microbial compounds and pre-existing drugs were screened for their ability to alter the intracellular d-aspartate level in mammalian cells, and several candidate compounds were identified. Detailed analytical studies suggested that two of these compounds, mithramycin A and geldanamycin, suppress the biosynthesis of d-aspartate in cells. Further studies suggested that these compounds act at distinct sites within the cell. These compounds may advance our current understanding of biosynthesis of d-aspartate in mammals, a whole picture of which remains to be disclosed.

Development of cerebral sulci and gyri in ferrets (Mustela putorius).

The present study aimed to clarify sulcation and gyration patterns in the developing cerebrum of ferrets. While the brain weight and fronto-occipital length of the cerebral hemisphere reached a plateau by postnatal day (PD) 42, the cerebral width reached a plateau at the rostral region by PD 21, and subsequently at the caudal region by PD 42. The ferret cerebrum already showed a convoluted surface with indentations of coronal and rostral suprasylvian sulci on PD 4. The presylvian and cruciate sulci emerged by PD 10, resulting in convolutions of gyri in the rostral half of the cerebrum. The caudal half of the cerebrum was infolded by the emergence of the pseudosylvian sulcus and the rhinal fissure by PD 10, and the caudal suprasylvian and lateral sulci by PD 21. The emergence of those sulci allowed a gyration in the caudal half of the cerebrum. Sexual differences in sulcation were detected by a more distinct convolution of the visual cortex in males than in females on PD 90. Those results, therefore, suggest that the ferret cerebrum experiences cortical maturation with sulcation and gyration in a rostrocaudal gradient manner. The present paper provides neuroanatomic references for normal development of cerebral sulci and gyri in both sexes of ferrets.