Calnexin mediates the maturation of GPI-anchors through ER retention

The protein folding and lipid moiety status of glycosylphosphatidylinositol-anchored proteins (GPI-APs) are monitored in the endoplasmic reticulum (ER), with calnexin playing dual roles in the maturation of GPI-APs. In the present study, we investigated the functions of calnexin in the quality control and lipid remodeling of GPI-APs in the ER. By directly binding the N-glycan on proteins, calnexin was observed to efficiently retain GPI-APs in the ER until they were correctly folded. In addition, sufficient ER retention time was crucial for GPI-inositol deacylation, which is mediated by post-GPI attachment protein 1 (PGAP1). Once the calnexin/calreticulin cycle was disrupted, misfolded and inositol-acylated GPI-APs could not be retained in the ER and were exposed on the plasma membrane. In calnexin/calreticulin-deficient cells, endogenous GPI-anchored alkaline phosphatase was expressed on the cell surface, but its activity was significantly decreased. ER stress induced surface expression of misfolded GPI-APs, but proper GPI-inositol deacylation occurred due to the extended time that they were retained in the ER. Our results indicate that calnexin-mediated ER quality control systems for GPI-APs are necessary for both protein folding and GPI-inositol deacylation.     

On the 710 nm fluorescence emitted by the diatom Phaeodactylum tricornutum at room temperature

The fluorescence emitted at 710 nm by Phaeodactylum tricornutum (F(710)) was characterized. Development of F(710) was found to be regulated by the quality of light needed for algal growth: weak red light absorbed mainly by Chl a induced its development, and weak blue-green light absorbed mainly by fucoxanthin and Chl c suppressed it. The difference spectra between cells grown under the two light conditions revealed two Chl a forms, absorption peaks of which were located at 692 nm (Chl a(692)) and at 703 nm (Chl a(703)), respectively, in red-light-grown cells. During cell growth under red light, the appearance and intensification of the emission correlated well with development of Chl a(692) and Chl a(703) suggesting that the two forms of Chl a are involved in the energy flow to F(710). A clear induction phenomenon characteristic of the PSII fluorescence was observed not only with the emission at 680 nm but also with F(710), indicating that F(710) is emitted by PSII Chl a. Development of F(710) under red light was sensitive to cycloheximide, indicating that the development of the energy flow to F(710) requires protein synthesis and that the emitter is installed in a protein encoded in the nuclear genome like the light-harvesting complex (LHC). Centrifugal fractionation of pigment-protein complexes revealed F(710) to be located at fractions slightly heavier than the major LHC. Development of F(710) was also found in red-light-grown cells of the diatom Nitzschia closterium.     

Incidence of Prevotella intermedia and Prevotella nigrescens in Periodontal Health and Disease

The incidence of black-pigmented rods (BPRs), especially Prevotella intermedia and Prevotella nigrescens, in periodontal health and disease were examined. Furthermore, the degradative enzyme activities of P. intermedia were compared among the strains from periodontal health and disease. Microbiological specimens were collected from subgingival crevice or periodontal pocket by paper point. The BPRs were found in 71.1% of periodontally healthy subjects (n = 45), and in 47.1% of healthy sites (n = 34) and 87.8% of active sites (n = 41) among periodontally diseased patients. Porphyromonas gingivalis was detected only in active sites of periodontally diseased patients (17.8% of 180 strains). P. intermedia was the predominant BPR in both healthy and active sites (37.3 and 41.7%, respectively) of the patients. However, P. nigrescens was the predominant BPR (70.5% of 173 strains) in periodontally healthy subjects. The enzyme activities of esterase, esterase-lipase, acid-phosphatase and α-fucosidase of P. intermedia strains isolated from active sites in patients were significantly higher (P < 0.05) than those of healthy subjects. The results suggest that P. intermedia might increase the activity of degradative enzymes under a certain condition and support the progression of periodontitis.     

Temporal change in nutrient concentrations and phytoplankton biomass in short time scale local upwelling around the Izu Peninsula, Japan*

Temporal changes in nitrate plus nitrite and chlorophyll concentrationswere observed by continuous shipboard measurements in upwelledand coastal water masses covering an area 20 x 30 km² alongthe Izu Peninsula between May 22 and 26, 1982. Two localizedupwelling events were observed, one which had occurred beforeMay 22 and the other on May 26. Nutrient input into the surfacefrom upwelling and subsequent phytoplankton growth were clearlydocumented. Phytoplankton growth, estimated from the disappearanceof nutrients, agreed well with modelling estimates based uponsimulated culture experiments done previously in the same area.Phytoplankton growth terminated when nutrients were depleted.Similar stimulation of phytoplankton growth supported by localizedupwelling can be expected in other coastal regions where similarconditions exist.     

BMP-4 and retinoic acid synergistically induce activation of caspase-9 and cause apoptosis of P19 embryonal carcinoma cells cultured as a monolayer

In monolayer cultures of P19 EC cells treated with both all-trans retinoic acid (RA) and bone morphogenetic protein (BMP)-4 (RA/BMP-4 treatment), many non-adherent apoptotic cells and activated caspase-3-positive cells were observed, but they were not observed in cells treated with RA or BMP-4 alone. Consistent with the appearance of activated caspase-3-positive cells, BMP-4 and RA together induced processing of caspase-9, Ac-DEVD-MCA cleavage activity and DNA fragmentation. These three activities were observed infrequently or not at all when cells were treated with RA or BMP-4 alone. In the RA/BMP-4 treatment-induced apoptosis, caspase-9 was upstream of caspase-3 in the enzyme cascade, and the caspase-9 to -3 step was key in the apoptotic pathway. Bcl-xL inhibited processing of caspase-9, Ac-DEVD-MCA cleavage activity and DNA fragmentation induced by RA/BMP-4 treatment. However, unlike staurosporine-induced apoptosis, cytochrome c, which activates caspase-9, was not detected in the cytosol of RA/BMP-4-treated cells. RA and BMP-4 may activate caspase-9 through an apoptotic pathway other than the Apaf-1/cytochrome c pathway. The prominent decrease of X-chromosome-linked inhibitory apoptosis protein (XIAP) in the cytosol may explain the activation of caspase-9 induced by RA and BMP-4 treatment.     

Modeling and simulation of competition between two microorganisms for a single inhibitory substrate in a biofilm reactor

A simple biofilm model was developed to simulate the competition between two microorganisms for a common inhibitory substrate. The following assumptions were made for the simulations: (1) the biofilm has a uniform thickness and is composed of 5 segments, (2) growth of two microorganisms A and B which utilize the common substrate is expressed by the Haldane kinetics with a spatial limitation term and is independent of the other microorganism in the biofilm reactor, and (3) diffusion of the substrate, movement of the microorganisms, and continuous loss of the biomass by shearing are expressed by Fick's Law-type equations. The qualitative behavior of the biofilm reactor is characterized by five regions, I-V, depending on the operation conditions, the substrate concentration in feed, and the dilution rate. In region I, both microorganisms are washed out of the biofilm reactor. In region II, microorganism B is washed out, and in region III, microorganism A is washed out of the biofilm. In region IV, both microorganisms coexist with one another. In region V, both microorganisms coexist with a sustained oscillatory behavior. Convergence to regions I-V depends on the initial conditions. In regions II-V, washout of either or both microorganisms is also observed with initial conditions too far away.