Anion Transport or Nucleotide Binding by Ucp2 Is Indispensable for Ucp2-Mediated Efferocytosis

Rapid and efficient engulfment of apoptotic cells is an essential property of phagocytes for removal of the large number of apoptotic cells generated in multicellular organisms. To achieve this, phagocytes need to be able to continuously uptake apoptotic cells. It was recently reported that uncoupling protein 2 (Ucp2) promotes engulfment of apoptotic cells by increasing the phagocytic capacity, thereby allowing cells to continuously ingest apoptotic cells. However, the functions of Ucp2, beyond its possible role in dissipating the mitochondrial membrane potential, that contribute to elevation of the phagocytic capacity have not been determined. Here, we report that the anion transfer or nucleotide binding activity of Ucp2, as well as its dissipation of the mitochondrial membrane potential, is necessary for Ucp2-mediated engulfment of apoptotic cells. To study these properties, we generated Ucp2 mutations that affected three different functions of Ucp2, namely, dissipation of the mitochondrial membrane potential, transfer of anions, and binding of purine nucleotides. Mutations of Ucp2 that affected the proton leak did not enhance the engulfment of apoptotic cells. Although anion transfer and nucleotide binding mutations did not affect the mitochondrial membrane potential, they exerted a dominant-negative effect on Ucp2-mediated engulfment. Furthermore, none of our Ucp2 mutations increased the phagocytic capacity. We conclude that dissipation of the proton gradient by Ucp2 is not the only determinant of the phagocytic capacity and that anion transfer or nucleotide binding by Ucp2 is also essential for Ucp2-mediated engulfment of apoptotic cells.     

Determination of the time transferring cells for astaxanthin production considering two-stage process of Haematococcus pluvialis cultivation

The two-stage culture system consisting of green vegetative growth and reddish inductive production stages has been widely accepted for the production of astaxanthin using Haematococcuspluvialis. However, little has been known about the appropriate cellular phase of H.pluvialis for transferring into the astaxanthin inductive conditions. In this study, we determined the optimal growth phase of H.pluvialis for transferring into the second production stage. Astaxanthin productivities were correlated with growth phases, as senescent green phases could increase more than 10-fold greater than juvenile green phases. Our results clearly demonstrated the appropriateness of the senescent vegetable cells for transferring into the production stage, due to the increased capacity to accumulate astaxanthin.     

Response of nitrifying bacterial communities to the increased thiocyanate concentration in pre-denitrification process

Changes in process performance and the nitrifying bacterial community associated with an increase of thiocyanate (SCN⁻) loading were investigated in a pre-denitrification process treating industrial wastewater. The increased SCN⁻ loading led to the concentration of total nitrogen (TN) in the final effluent, but increasing the internal recycling ratio as an operation parameter from 2 to 5 resulted in a 21% increase in TN removal efficiency. In the aerobic reactor, we found that the Nitrosomonas europaea lineage was the predominant ammonia oxidizing bacteria (AOB) and the percentages of the AOB population within the total bacteria increased from about 4.0% to 17% with increased SCN⁻ concentration. The increase of nitrite loading seemed to change the balance between Nitrospira and Nitrobacter, resulting in the high dominance of Nitrospira over Nitrobacter. Meanwhile, a Thiobacillus thioparus was suggested to be the main microorganism responsible for the SCN⁻ biodegradation observed in the system.     

Synthesis and evaluation of cytotoxic effects of hanultarin and its derivatives

One of the known cytotoxic lignans is (-)-1-O-feruloyl-secoisolariciresinol designated as hanultarin, which was isolated from the seeds of Trichosanthes kirilowii. In this Letter, we described the first synthesis of 1-O-feruloyl-secoisolariciresinol, 1,4-O-diferuloyl-secoisolariceresinol and their analogues. The cytotoxicities of these compounds were evaluated against several cancer cell lines. Interestingly, we found that the feruloyl diester derivative of secoisolariciresinol was the most active cytotoxic compound against all the cancer cells tested in this experiment. The IC(50) values of the1,4-O-diferuloyl-secoisolariceresinol were in the range of 7.1-9.8μM except one cell line. In considering that both ferulic acid and secoisolariciresinol are commonly found in many plants and have no cytotoxicity, this finding is remarkable in that simple covalent bonds between the ferulic acid and secoisolariciresinol cause a cytotoxic effect.     

A single, low, oral dose of a 5-carbon-linked trioxane dimer orthoester plus mefloquine cures malaria-infected mice

Four 5-carbon-linked trioxane dimer orthoesters (6a-6d) have been prepared in 4 or 5 chemical steps from the natural trioxane artemisinin (1). When administered orally to malaria-infected mice using a single dose of only 6 mg/kg body weight along with 18 mg/kg of mefloquine hydrochloride, trioxane dimer orthoester sulfone 6d completely and safely cured the mice; after 30 days, the cured mice showed no detectable parasitemia, gained at least as much weight as the control mice (no infection), and behaved normally.     

Large-scale plantlets conversion from cotyledonary somatic embryos of Kalopanax septemlobus tree using bioreactor cultures

A suitable bioreactor system for large scale embryo-to-plantlets conversion of Kalopanax septemlobus was established. In temporary immersion with net (TIN) bioreactor, 85% of embryos successfully produced plantlets whereas in continuous immersion with net (CIN) bioreactor, only conversion rate of 29.3% was obtained. Embryos cultured in TIN bioreactor produced more vigorous plantlets in terms of fresh weight, height, root length, roots and leaves quantity. In CIN bioreactor, Kalopanax plantlets showed high malondialdehyde (MDA) content and increased activities of reactive oxygen species (ROS)-processing enzymes, such as ascorbate peroxidase (APX) and glutathione reductase (GR) indicating the occurrence of oxidative stress. However, superoxide dismutase (SOD) and catalase (CAT) showed similar activities in plantlets grown in different bioreactors. Kalopanax plantlets grown in both TIN and CIN bioreactors were harvested and transferred to greenhouse for their acclimatization. Plantlets grown in CIN bioreactor exhibited low survival rate (75.8%) compared to those grown in TIN bioreactor (100%). MDA content decreased with progression of acclimatization indicating a decrease in oxidative stress. However, MDA level in CIN derived plantlets was higher than TIN derived plantlets. In TIN derived plantlets, an increase in SOD and GR activities were observed after 1 week and thereafter decreased. CAT activity decreased while APX activity started to increase after 1 week of acclimatization. The results indicated that Kalopanax plantlets were able to overcome oxidative stress mainly through SOD activity. However, levels of antioxidant enzyme activities were higher in CIN derived plantlets than TIN derived plantlets. Kalopanax plantlets obtained from TIN bioreactor performed better during the acclimatization phase and showed higher survival rate than material obtained on CIN bioreactor or conventional culture systems.     

C<Subscript>6</Subscript>-ceramide enhances phagocytic activity of Kupffer cells through the production of endogenous ceramides

Ceramide has been suggested to be not only a tumorsuppressive lipid but also a regulator of phagocytosis. We examined whether exogenous cell-permeable C₆-ceramide enhances the phagocytic activity of Kupffer cells (KCs) and affects the level of cellular ceramides. Rat KCs were isolated by collagenase digestion and differential centrifugation, using Percoll system. Phagocytic activity was measured by FACS analysis after incubating KCs with fluorescence-conjugated latex beads, and the level of cellular ceramide was analyzed by liquid chromatography tandem-mass spectrometry (LC-MS/MS). In this study we found that permeable C₆-ceramide increases the cellular levels of endogenous ceramides via a sphingosine-recycling pathway leading to enhanced phagocytosis by KCs.