Taming chlorophylls by early eukaryotes underpinned algal interactions and the diversification of the eukaryotes on the oxygenated Earth.

Extant eukaryote ecology is primarily sustained by oxygenic photosynthesis, in which chlorophylls play essential roles. The exceptional photosensitivity of chlorophylls allows them to harvest solar energy for photosynthesis, but on the other hand, they also generate cytotoxic reactive oxygen species. A risk of such phototoxicity of the chlorophyll must become particularly prominent upon dynamic cellular interactions that potentially disrupt the mechanisms that are designed to quench photoexcited chlorophylls in the phototrophic cells. Extensive examination of a wide variety of phagotrophic, parasitic, and phototrophic microeukaryotes demonstrates that a catabolic process that converts chlorophylls into nonphotosensitive 132,173-cyclopheophorbide enols (CPEs) is phylogenetically ubiquitous among extant eukaryotes. The accumulation of CPEs is identified in phagotrophic algivores belonging to virtually all major eukaryotic assemblages with the exception of Archaeplastida, in which no algivorous species have been reported. In addition, accumulation of CPEs is revealed to be common among phototrophic microeukaryotes (i.e., microalgae) along with dismantling of their secondary chloroplasts. Thus, we infer that CPE-accumulating chlorophyll catabolism (CACC) primarily evolved among algivorous microeukaryotes to detoxify chlorophylls in an early stage of their evolution. Subsequently, it also underpinned photosynthetic endosymbiosis by securing close interactions with photosynthetic machinery containing abundant chlorophylls, which led to the acquisition of secondary chloroplasts. Our results strongly suggest that CACC, which allowed the consumption of oxygenic primary producers, ultimately permitted the successful radiation of the eukaryotes throughout and after the late Proterozoic global oxygenation.

Profiling physicochemical and planktonic features from discretely/continuously sampled surface water.

There is an increasing need for assessing aquatic ecosystems that are globally endangered. Since aquatic ecosystems are complex, integrated consideration of multiple factors utilizing omics technologies can help us better understand aquatic ecosystems. An integrated strategy linking three analytical (machine learning, factor mapping, and forecast-error-variance decomposition) approaches for extracting the features of surface water from datasets comprising ions, metabolites, and microorganisms is proposed herein. The three developed approaches can be employed for diverse datasets of sample sizes and experimentally analyzed factors. The three approaches are applied to explore the features of bay water surrounding Odaiba, Tokyo, Japan, as a case study. Firstly, the machine learning approach separated 681 surface water samples within Japan into three clusters, categorizing Odaiba water into seawater with relatively low inorganic ions, including Mg, Ba, and B. Secondly, the factor mapping approach illustrated Odaiba water samples from the summer as rich in multiple amino acids and some other metabolites and poor in inorganic ions relative to other seasons based on their seasonal dynamics. Finally, forecast-error-variance decomposition using vector autoregressive models indicated that a type of microalgae (Raphidophyceae) grows in close correlation with alanine, succinic acid, and valine on filters and with isobutyric acid and 4-hydroxybenzoic acid in filtrate, Ba, and average wind speed. Our integrated strategy can be used to examine many biological, chemical, and environmental physical factors to analyze surface water.

Leaching of Metals and Metalloids from Hydrothermal Ore Particulates and Their Effects on Marine Phytoplankton.

Seafloor massive sulfide deposits have attracted much interest as mineral resources. Therefore, the potential environmental impacts of full-scale mining should be considered. In this study, we focused on metal and metalloid contamination that could be triggered by accidental leakage and dispersion of hydrothermal ore particulates from mining vessels into surface seawater. We determined the leaching potential of metals and metalloids from four hydrothermal ores collected from the Okinawa Trough into aerobic seawater and then evaluated the toxic effects of ore leachates on a phytoplankton species, Skeletonema marinoi-dohrnii complex, which is present ubiquitously in the ocean. Large amounts of metals and metalloids were released from the ground hydrothermal ores into seawater within 5 min under aerobic conditions. The main components of leachates were Zn + Pb, As + Sb, and Zn + Cu, which were obtained from the Fe-Zn-Pb-rich and Zn-Pb-rich zero-age, Ba-rich, and Fe-rich ores, respectively. The leachates had different chemical compositions from those of the ore. The rapid release and difference in chemical compositions between the leachates and the ores indicated that substances were not directly dissolved from the sulfide-binding mineral phase but from labile phases mainly on the adsorption-desorption interface of the ores under these conditions. All ore leachates inhibited the growth of S. marinoi-dohrnii complex but with different magnitudes of toxic effects. These results indicate that the fine particulate matter of hydrothermal ores is a potential source of toxic contamination that may damage primary production in the ocean. Therefore, we insist on the necessity for the prior evaluation of toxic element leachability from mineral ores into seawater to minimize mining impacts on the surface environment.

L-positioned Perforator Propeller Flap for Partial Breast Reconstruction with Axillary Dead Space.

Partial breast reconstruction using perforator flaps harvested from the lateral chest wall has become a well-established surgical technique recently. In the case of a partial mastectomy with an axillary lymph node dissection, there are 2 main defects; one is a partial breast defect and the other is an axillary dead space. To reconstruct the 2 separate defects with local flaps, basically 2 different flaps are needed, and usually, it is rather difficult to harvest 2 different local flaps in the adjacent area. To resolve this problem, we introduce the L-positioned perforator propeller flap (PPF). We used an L-positioned PPF on 2 female patients, aged 46 and 47 years old, who were suffering from breast cancer in the upper outer quadrant. The concept of this flap design is as follows: the partial breast defect is reconstructed with the longer lobe of the L-positioned PPF and the axillary defect is filled with the smaller lobe of the L-positioned PPF at the same time. The reconstruction time was 2 hours and 0 minutes and 1 hour and 46 minutes in each case. The patients were successfully provided with aesthetically acceptable breast reconstruction without postoperative complications. Moreover, both patients had consecutive postoperative radiotherapy on the reconstructed area without complications. With this flap design, it is possible for patients to have safe and aesthetic reconstruction with only 1 local flap and fewer invasive procedures.

Carotenogenesis diversification in phylogenetic lineages of Rhodophyta.

Carotenoid composition is very diverse in Rhodophyta. In this study, we investigated whether this variation is related to the phylogeny of this group. Rhodophyta consists of seven classes, and they can be divided into two groups on the basis of their morphology. The unicellular group (Cyanidiophyceae, Porphyridiophyceae, Rhodellophyceae, and Stylonematophyceae) contained only ß-carotene and zeaxanthin, "ZEA-type carotenoids." In contrast, within the macrophytic group (Bangiophyceae, Compsopogonophyceae, and Florideophyceae), Compsopogonophyceae contained antheraxanthin in addition to ZEA-type carotenoids, "ANT-type carotenoids," whereas Bangiophyceae contained α-carotene and lutein along with ZEA-type carotenoids, "LUT-type carotenoids." Florideophyceae is divided into five subclasses. Ahnfeltiophycidae, Hildenbrandiophycidae, and Nemaliophycidae contained LUT-type carotenoids. In Corallinophycidae, Hapalidiales and Lithophylloideae in Corallinales contained LUT-type carotenoids, whereas Corallinoideae in Corallinales contained ANT-type carotenoids. In Rhodymeniophycidae, most orders contained LUT-type carotenoids; however, only Gracilariales contained ANT-type carotenoids. There is a clear relationship between carotenoid composition and phylogenetics in Rhodophyta. Furthermore, we searched open genome databases of several red algae for references to the synthetic enzymes of the carotenoid types detected in this study. ß-Carotene and zeaxanthin might be synthesized from lycopene, as in land plants. Antheraxanthin might require zeaxanthin epoxydase, whereas α-carotene and lutein might require two additional enzymes, as in land plants. Furthermore, Glaucophyta contained ZEA-type carotenoids, and Cryptophyta contained ß-carotene, α-carotene, and alloxanthin, whose acetylenic group might be synthesized from zeaxanthin by an unknown enzyme. Therefore, we conclude that the presence or absence of the four enzymes is related to diversification of carotenoid composition in these three phyla.

Taxonomic study of a new eustigmatophycean alga, Vacuoliviride crystalliferum gen. et sp. nov.

This study investigated the taxonomic affiliation of the algal strain nak-9, which has been reported to absorb radioactive cesium with high efficiency, using light and electron microscopy, and molecular phylogenetic analysis based on 18S ribosomal RNA gene (rDNA) sequences. This alga is spherical and coccoid, with a smooth cell wall, large vacuole, crystalline structure, reddish globule, and refractile granules (lamellate vesicles). The cells possess one to several greenish parietal chloroplasts with a bulging pyrenoid surrounded by lamellate vesicles. The chloroplasts include orderly thylakoid lamellae but no girdle lamella. Molecular phylogenetic analysis suggests that strain nak-9 is a member of the eustigmatophycean clade, which includes Goniochloris, Pseudostaurastrum, and Trachydiscus. On the basis of these results, we propose that strain nak-9 (NIES-2860) comprises a new species and new genus of the Eustigmatophyceae, Vacuoliviride crystalliferum gen. et sp. nov.

Phylogeny, ultrastructure, and flagellar apparatus of a new Marimonad flagellate Abollifer globosa sp. nov. (Imbricatea, Cercozoa).

Abollifer is a little-known genus of marine heterotrophic flagellates with no ultrastructural and molecular information, and its taxonomic position remains uncertain. In this study, we report a new species of Abollifer, Abollifer globosa sp. nov., isolated from a seawater sample collected at Tokyo Bay. To reveal the taxonomic position and morphological characteristics of A. globosa, we performed light and electron microscopic observations and a phylogenetic analysis using small subunit ribosomal DNA sequences. A. globosa cells were 29.5 µm in length and 22.4 µm in width, oval or ovoid in shape with an apical projection. Two unequal flagella emerged from a deep subapical flagellar pit. The rim of the flagellar pit except for the ventral side was swollen. Electron microscopic observations showed that A. globosa possessed mitochondria with tubular cristae, a Golgi apparatus, microbodies, extrusomes, and many endosymbiotic bacteria. Basal bodies were arranged in parallel. The flagellar apparatus of A. globosa showed affinity with common gliding cercozoan flagellates. Our phylogenetic tree showed that A. globosa branched as the sister position of order Marimonadida (Imbricatea, Cercozoa). On the basis of the morphological and molecular phylogenetic analysis, we conclude that A. globosa is a new member of the order Marimonadida.

Global searches for microalgae and aquatic plants that can eliminate radioactive cesium, iodine and strontium from the radio-polluted aquatic environment: a bioremediation strategy.

The Fukushima 1 Nuclear Power Plant accident in March 2011 released an enormously high level of radionuclides into the environment, a total estimation of 6.3 × 10¹7 Bq represented by mainly radioactive Cs, Sr, and I. Because these radionuclides are biophilic, an urgent risk has arisen due to biological intake and subsequent food web contamination in the ecosystem. Thus, urgent elimination of radionuclides from the environment is necessary to prevent substantial radiopollution of organisms. In this study, we selected microalgae and aquatic plants that can efficiently eliminate these radionuclides from the environment. The ability of aquatic plants and algae was assessed by determining the elimination rate of radioactive Cs, Sr and I from culture medium and the accumulation capacity of radionuclides into single cells or whole bodies. Among 188 strains examined from microalgae, aquatic plants and unidentified algal species, we identified six, three and eight strains that can accumulate high levels of radioactive Cs, Sr and I from the medium, respectively. Notably, a novel eustigmatophycean unicellular algal strain, nak 9, showed the highest ability to eliminate radioactive Cs from the medium by cellular accumulation. Our results provide an important strategy for decreasing radiopollution in Fukushima area.

13²,17³-Cyclopheophorbide b enol as a catabolite of chlorophyll b in phycophagy by protists.

Both 13(2),17(3)-cyclopheophorbide a and b enols were produced along with ingestion of green microalgae containing chlorophylls a and b by a centrohelid protist (phycophagy). The results suggest that chlorophyll b as well as chlorophyll a were directly degraded to colored yet non-phototoxic catabolites in the protistan phycophagic process. Such a simple process by the predators makes a contrast to the much sophisticated chlorophyll degradation process of land plants and some algae, where phototoxicity of chlorophylls was cancelled through the multiple enzymatic steps resulting in colorless and non-phototoxic catabolites.

[Experiences of caring for patients with lymphedema and dementia].

Recently, the importance of prevention and care of lymphedema patients has been recognized. To improve edema reduction and skin condition, continued care is necessary. However, everyday care requires time and effort, and patients may neglect the required routine. Therefore, it is necessary for patients to recognize the importance of lymphedema and acquire the knowledge and skills to care for their condition. In the case of patients with dementia, it is important to ensure 1) comfortable care for the patients, 2) flexibility of care according to the physical and mental state of the patients, and 3) education of visiting home care staff.

Ubiquity and quantitative significance of detoxification catabolism of chlorophyll associated with protistan herbivory.

Chlorophylls are essential components of the photosynthetic apparati that sustain all of the life forms that ultimately depend on solar energy. However, a drawback of the extraordinary photosensitizing efficiency of certain chlorophyll species is their ability to generate harmful singlet oxygen. Recent studies have clarified the catabolic processes involved in the detoxification of chlorophylls in land plants, but little is understood about these strategies in aquatic ecosystem. Here, we report that a variety of heterotrophic protists accumulate the chlorophyll a catabolite 13(2),17(3)-cyclopheophorbide a enol (cPPB-aE) after their ingestion of algae. This chlorophyll derivative is nonfluorescent in solution, and its inability to generate singlet oxygen in vitro qualifies it as a detoxified catabolite of chlorophyll a. Using a modified analytical method, we show that cPPB-aE is ubiquitous in aquatic environments, and it is often the major chlorophyll a derivative. Our findings suggest that cPPB-aE metabolism is one of the most important, widely distributed processes in aquatic ecosystems. Therefore, the herbivorous protists that convert chlorophyll a to cPPB-aE are suggested to play more significant roles in the modern oceanic carbon flux than was previously recognized, critically linking microscopic primary producers to the macroscopic food web and carbon sequestration in the ocean.

α-Carotene and its derivatives have a sole chirality in phototrophic organisms?

Carotenoids in eukaryotic phototrophic organisms can be classified into two groups; ß-carotene and its derivatives, and α-carotene and its derivatives. We re-examined distribution of α-carotene and its derivatives among various taxa of aquatic algae (17 classes) and land plants. α-carotene and its derivatives were found from Rhodophyceae (macrophytic type), Cryptophyceae, Euglenophyceae, Chlorarachniophyceae, Prasinophyceae, Chlorophyceae, Ulvophyceae, Charophyceae, and land plants, while they could not be detected from Glaucophyceae, Rhodophyceae (unicellular type), Chryosophyceae, Raphidophyceae, Bacillariophyceae, Phaeophyceae, Xanthophyceae, Eustigmatophyceae, Haptophyceae, and Dinophyceae. We also analyzed the chirality of α-carotene and/or its derivatives, such as lutein and siphonaxanthin, and found all of them had only (6'R)-type, not (6'S)-type.

Transforming growth factor ß and Ras/MEK/ERK signaling regulate the expression level of a novel tumor suppressor Lefty.

OBJECTIVES: The objectives of the present study were (i) to identify a novel tumor suppressor gene whose expression level was regulated by transforming growth factor (TGF-ß) and (ii) to evaluate the effect of Ras/MEK/ERK signaling on TGF-ß-dependent Lefty up-regulation. METHODS: Human pancreatic cancer cell lines were used. The effect of Ras/MEK/ERK pathway on TGF-ß-mediated Lefty up-regulation was tested by adding K-ras small interfering RNA, MEK inhibitor U0126, or extracellular signal-regulated kinase (ERK) inhibitor LY294002. RESULTS: Transforming growth factor ß upregulated Lefty messenger RNA levels within 6 of the 7 cell lines. Lefty exerts an antagonistic effect against the tumor-promoting molecule, Nodal, as recombinant Lefty suppressed Nodal-mediated proliferation. Interestingly, inhibition of the Ras/MEK/ERK pathway dramatically enhanced TGF-mediated Lefty up-regulation, suggesting that Ras/MEK/ERK signaling suppresses TGF-ß-Lefty pathway. CONCLUSIONS: Our data suggest that Lefty is a novel TGF-ß target molecule that mediates growth inhibition of pancreatic cancer cells. In addition, activation of the Ras/MEK/ERK pathway serves as a mechanism by which pancreatic cancer escapes from growth inhibition by the TGF-ß-Lefty axis. The results imply a novel therapeutic strategy for pancreatic cancer, that is, combination treatment with Ras/MEK/ERK inhibitors and TGF-ß.

[A case of an end-of-life cancer patient in which lymphedema care improved the QOL enabling home hospice care].

Recently, the importance of prevention and care of lymphedema has been recognized. It appears that the complex physical therapy is relatively taboo for an end-of-life cancer patient with lymphedema. Therefore, an intervention of care is often difficult. In our case, the patient strongly desired a hospice care at home, so we introduced a home hospice care gradually taking a step by step approach with anticipated harmful symptoms. As a result, the home hospice care was doable with ADL and QOL improvements. For lymphedema care of an end-of-life cancer patient, a goal setting, sharing the patient's will based on anticipated symptoms, and an introduction of a gradual care were all important.

EVOLUTIONARY ANALYSES OF THE NUCLEAR-ENCODED PHOTOSYNTHETIC GENE psbO FROM TERTIARY PLASTID-CONTAINING ALGAE IN DINOPHYTA(1).

Although the dinophytes generally possess red-algal-derived secondary plastids, tertiary plastids originating from haptophyte and diatom ancestors are recognized in some lineages within the Dinophyta. However, little is known about the nuclear-encoded genes of plastid-targeted proteins from the dinophytes with diatom-derived tertiary plastids. We analyzed the sequences of the nuclear psbO gene encoding oxygen-evolving enhancer protein from various algae with red-algal-derived secondary and tertiary plastids. Based on our sequencing of 10 new genes and phylogenetic analysis of PsbO amino acid sequences from a wide taxon sampling of red algae and organisms with red-algal-derived plastids, dinophytes form three separate lineages: one composed of peridinin-containing species with secondary plastids, and the other two having haptophyte- or diatom-derived tertiary plastids and forming a robust monophyletic group with haptophytes and diatoms, respectively. Comparison of the N-terminal sequences of PsbO proteins suggests that psbO genes from a dinophyte with diatom-derived tertiary plastids (Kryptoperidinium) encode proteins that are targeted to the diatom plastid from the endosymbiotic diatom nucleus as in the secondary phototrophs, whereas the fucoxanthin-containing dinophytes (Karenia and Karlodinium) have evolved an additional system of psbO genes for targeting the PsbO proteins to their haptophyte-derived tertiary plastids from the host dinophyte nuclei.

[Two cases of Stewart-Treves syndrome observed at Sanshu hospital].

Stewart-Treves syndrome is angiosarcoma, which occurs in the chronic lymphedema of the upper or lower limbs. Presently, an effective therapy is not established. The survival period is only several months to one year. There are some reports that angiosarcoma will occur in the period of 5~15 years after a continuation of lymphedema of the extremities. Therefore, it is important to prevent lymphedema after surgical operations of primary disease and the lymphedema.

Anticancer effects of a tertiary sulfonium compound, dimethylsulfoniopropionate, in green sea algae on Ehrlich ascites carcinoma-bearing mice.

The saline and dimethylsulfoniopropionate (DMSP) solutions at 5, 10 and 20 mM were preliminarily injected intraperitoneally every other day into two control and three DMSP groups of mice (n=8) for 2 wk and thereafter Ehrlich ascites-carcinoma (EAC) cells were peritoneally injected to one control and three DMSP groups of mice, leaving one control group without the EAC injection. Then, the body weight and survival time of all mice were examined over a long rearing time up to 300 d. All EAC-bearing mice, especially the carcinoma control and 5 mM DMSP-carcinoma group mice, rapidly increased their body weights early and then died by day 50 and day 90, respectively. In contrast, the administration of 10 and 20 mM DMSP solutions prolonged the lives of EAC-bearing mice at the survival rate of 50 and 63% respectively up to 300 d without any side effects. Furthermore, the administration of 10 mM DMSP solution proved to activate the delayed-type hypersensitivity of EAC bearing-mice, and the DMSP solutions over the concentrations of 5 to 30 mM to slightly reduce the dead cells in EAC cells on the synthetic medium. Accordingly, the preliminary supplementation of 10 and 20 mM DMSP solutions to EAC-bearing mice was proven to maintain their lives at high survival rates without direct damage to EAC cells for a long time, probably due to the activation of the immune system without any side effects.

[Dysfunction of multiple cranial nerves in cephalic tetanus--case report].

A 77-year-old man non-immunized to tetanus suffered head trauma on the right side when he tumbled from a height of approximately 2m. Five days later, he experienced difficulty in opening his mouth and developed right ptosis. He was referred to our hospital 2 days post-ictus. The patient suffered trismus, and developed right Horner's syndrome with in a week. Symptoms due to multiple cranial nerve palsies were observed: right inferior oblique muscle weakness, reduced right corneal reflex, right facial palsy, dysphagia, and abnormal tongue movements. Neuroimages (computed tomography, magnetic resonance imaging, and angiography) of the basal skull and internal carotid arteries revealed no abnormalities. From the symptoms associated with his infected head wound and clinical follow-up, we suggested that he had cephalic tetanus. We subsequently conducted the following treatments: debridement of the wound, intravenous infusion of antitetanus human immunoglobulin (AHI), intrathecal AHI infusion, and systemic administration of benzylpenicillin. His condition improved with these treatments, and without any complications such as autonomic nervous system dysfunction or classical tetanic spasms. This case suggests that we should consider the possibility of cephalic tetanus when we observe a patient with cranial nerve palsy associated with injury.

New mechanism of transforming growth factor-beta signaling in hepatoma: Dramatic up-regulation of tumor initiating cells and epidermal growth factor receptor expression.

AIM: Transforming growth factor-beta (TGF-beta) has dual activity in tumor cells. We studied the effect of TGF-beta on tumor-initiating cells (TICs), which are similar in self-renewal and differentiation features to normal adult stem cells. METHODS: We used side population (SP) cells that exclude DNA binding dye Hoechst 33342 to obtain TICs, studied the differences in the kinetics of the SP cell response to TGF-beta treatment between hepatic tumor cell lines, and performed gene analysis. RESULTS: SP cells from all cell lines have higher proliferative ability compared to non-SP cells and they are drug resistant. TGF-beta treatment increased the percentage of SP cells (%SP) and the survival rate; chemotherapeutic drug resistance developed only in K-251 SP cells. Gene analysis showed that TGF-beta up-regulated epidermal growth factor receptor (EGFR) only in K-251 cells. There were no EGFR mutations in K-251, which had been reported in lung cancer. Knockdown of Smad4 using the small-interfering RNA technique in K-251 cells inhibited EGFR overexpression and significantly decreased the %SP. In contrast, the JNK inhibitor had little effect on EGFR expression or the %SP. CONCLUSION: TGF-beta treatment of K-251 cells causes tumor progression and the anti-cancer drug resistant phenotype by increasing SP.