Trace elements influenced by environmental changes in Lake Biwa: (II) Chemical variations in the hypolimnion over the last half-century

The history of the deep north basin of Lake Biwa extends over 430,000 years. Although it has probably been oxic and oligotrophic since its formation, human impacts have been changing lake conditions. In this paper, we discuss long-term changes in the chemistry of bottom water by compiling literature and through our own data over the last half-century. Long-term records show an increase in temperature, decrease in dissolved oxygen (DO), and increase in nutrients in bottom water. The stoichiometry among oxygen and nutrients indicates that changes are basically consistent with aerobic decomposition of organic matter. These changes are most likely the result of global warming and local eutrophication. Of particular note, yearly minimum DO concentrations <50 µmol kg^?1 have started to occur frequently at ~90 m depth since 1999. Manganese (Mn) concentrations in bottom water are at their minimum during the turnover period and at a maximum during the late stratification period each year. Yearly minimum Mn concentration has been within a narrow range over the last 30 years (0.25 ± 0.07 µmol kg^?1, n = 12). However, abnormally high Mn concentrations (up to 9.3 µmol kg^?1) were observed in 2007, caused by reductive release of a substantial amount of Mn from suboxic sediments and subsequent oxidation in bottom water. The concentration of arsenic (As) has gradually increased over the last 20 years in a similar manner, with a homologous element of phosphorus (P), resulting in an observed range of 17–29 nmol kg^?1 in 2010. The accumulation rate was ~0.8 nmol kg^?1 year^?1 for As and ~6 nmol kg^?1 year^?1 for P.     

Various Effects of Sandblasting of Dental Restorative Materials

Background

Sandblasting particles which remain on the surfaces of dental restorations are removed prior to cementation. It is probable that adhesive strength between luting material and sandblasting particle remnants might exceed that with restorative material. If that being the case, blasting particles adhere to sandblasted material surface could be instrumental to increasing adhesive strength like underlying bonding mechanism between luting material and silanized particles of tribochemical silica coating-treated surface. We hypothesize that ultrasonic cleaning of bonding surfaces, which were pretreated with sandblasting, may affect adhesive strength of a resin luting material to dental restorative materials.

Methods

We therefore observed adhesive strength of resin luting material to aluminum oxide was greater than those to zirconia ceramic and cobalt-chromium alloy beforehand. To measure the shear bond strengths of resin luting material to zirconia ceramic and cobalt-chromium alloy, forty specimens of each restorative material were prepared. Bonding surfaces were polished with silicon abrasive paper and then treated with sandblasting. For each restorative material, 40 sandblasted specimens were equally divided into two groups: ultrasonic cleaning (USC) group and non-ultrasonic cleaning (NUSC) group. After resin luting material was polymerized on bonding surface, shear test was performed to evaluate effect of ultrasonic cleaning of bonding surfaces pretreated with sandblasting on bond strength.

Results

For both zirconia ceramic and cobalt-chromium alloy, NUSC group showed significantly higher shear bond strength than USC group.

Conclusions

Ultrasonic cleaning of dental restorations after sandblasting should be avoided to retain improved bonding between these materials.     

Changing patterns of gene expression in dictyostelium prestalk cell subtypes recognized by in situ hybridization with genes from microarray analyses

We used microarrays carrying most of the genes that are developmentally regulated in Dictyostelium to discover those that are preferentially expressed in prestalk cells. Prestalk cells are localized at the front of slugs and play crucial roles in morphogenesis and slug migration. Using whole-mount in situ hybridization, we were able to verify 104 prestalk genes. Three of these were found to be expressed only in cells at the very front of slugs, the PstA cell type. Another 10 genes were found to be expressed in the small number of cells that form a central core at the anterior, the PstAB cell type. The rest of the prestalk-specific genes are expressed in PstO cells, which are found immediately posterior to PstA cells but anterior to 80% of the slug that consists of prespore cells. Half of these are also expressed in PstA cells. At later stages of development, the patterns of expression of a considerable number of these prestalk genes changes significantly, allowing us to further subdivide them. Some are expressed at much higher levels during culmination, while others are repressed. These results demonstrate the extremely dynamic nature of cell-type-specific expression in Dictyostelium and further define the changing physiology of the cell types. One of the signals that affect gene expression in PstO cells is the hexaphenone DIF-1. We found that expression of about half of the PstO-specific genes were affected in a mutant that is unable to synthesize DIF-1, while the rest appeared to be DIF independent. These results indicate that differentiation of some aspects of PstO cells can occur in the absence of DIF-1.     

Triggering of the vascular permeability reaction by activation of the Hageman factor-prekallikrein system by house dust mite proteinase

A 30-kilodalton (kDa) proteinase from the house dust mite Dermatophagoides farinae (Df-proteinase) was recently purified (Takahashi et al. (1990) Int. Arch. Allergy Appl. Immunol. 91, 80-85). In this paper we detailed the biological activities of the Df-proteinase. The activation of the kinin cascade by Df-proteinase was examined in vitro by using purified guinea pig Hageman factor (HF), prekallikrein (PK) and high-molecular-weight kininogen (HMWK) and the effect of this proteinase on endogenous human plasma proteinase inhibitors (serpins) and alpha 2-macroglobulin was tested. In addition, enhancement of the vascular permeability reaction in guinea pig skin by Df-proteinase was examined in vivo. These experiments showed that Df-proteinase could activate all the steps of the kinin-generating cascade, i.e., HF, PK and HMWK, and that Df-proteinase retained proteolytic activity even in the presence of an excess amount of endogenous proteinase inhibitors in plasma. We also found that the marked enhancement of the vascular permeability reaction was induced by Df-proteinase via the activation of the kinin-generating cascade without the release of histamine. From these results, we conclude that the proteinase of the house dust mite, Df-proteinase, has the potential to generate bradykinin and that the presence of this proteinase in biological systems would exacerbate inflammatory reactions in some pathological conditions.     

Phylogenetic analysis of diprotodontian marsupials based on complete mitochondrial genomes

Australidelphia is the cohort, originally named by Szalay, of all Australian marsupials and the South American Dromiciops. A lot of mitochondria and nuclear genome studies support the hypothesis of a monophyly of Australidelphia, but some familial relationships in Australidelphia are still unclear. In particular, the familial relationships among the order Diprotodontia (koala, wombat, kangaroos and possums) are ambiguous. These Diprotodontian families are largely grouped into two suborders, Vombatiformes, which contains Phascolarctidae (koala) and Vombatidae (wombat), and Phalangerida, which contains Macropodidae, Potoroidae, Phalangeridae, Petauridae, Pseudocheiridae, Acrobatidae, Tarsipedidae and Burramyidae. Morphological evidence and some molecular analyses strongly support monophyly of the two families in Vombatiformes. The monophyly of Phalangerida as well as the phylogenetic relationships of families in Phalangerida remains uncertain, however, despite searches for morphological synapomorphy and mitochondrial DNA sequence analyses. Moreover, phylogenetic relationships among possum families (Phalangeridae, Petauridae, Pseudocheiridae, Acrobatidae, Tarsipedidae and Burramyidae) as well as a sister group of Macropodoidea (Macropodidae and Potoroidae) remain unclear. To evaluate familial relationships among Dromiciops and Australian marsupials as well as the familial relationships in Diprotodontia, we determined the complete mitochondrial sequence of six Diprotodontian species. We used Maximum Likelihood analyses with concatenated amino acid and codon sequences of 12 mitochondrial protein genomes. Our analysis of mitochondria amino acid sequence supports monophyly of Australian marsupials+Dromiciops and monophyly of Phalangerida. The close relatedness between Macropodidae and Phalangeridae is also weakly supported by our analysis.     

The production-to-respiration ratio and its implication in Lake Biwa,Japan

Production-to-respiration (P:R) ratio was estimated at an offshore site of Lake Biwa in order to examine whether the plankton and benthic community is subsidized with allochthonous organic carbon, and to clarify the role of this lake as potential source or sink of carbon dioxide. The respiration rate of protozoan and metazoan plankton was calculated from their biomass and empirical equations of oxygen consumption rates, and that of bacterioplankton was derived from their production rate and growth efficiency. In addition, the carbon mineralization rate in the lake sediments was estimated from the accumulation rate of organic carbon, which was determined using a ²¹⁰Pb dating technique. On an annual basis, the sum of respiration rates of heterotrophic plankton was comparable to net primary production rate measured by the ¹³C method. However, when the mineralization rate in the lake sediments was included, the areal P:R ratio was 0.89, suggesting that Lake Biwa is net heterotrophic at the offshore site with the community being subsidized with allochthonous organic carbon. Such a view was supported by the surface water pCO₂ that was on average higher than that of the atmosphere. However, the estimate of net CO₂ release rate was close to that of carbon burial rate in the sediments. The result suggests that the role of Lake Biwa in relation to atmospheric carbon is almost null at the offshore site, although the community is supported partially by organic carbon released from the surrounding areas.     

Insulin regulates Na+/glucose cotransporter activity in rat small intestine

In order to examine the involvement of insulin in the activity of Na+/glucose cotransporter in rat small intestine, we compared Na(+)-dependent uptake of D-glucose by brush-border membrane vesicles prepared from control, streptozotocin-induced diabetic, insulin-treated diabetic and starved diabetic rats. In four groups, the uptake of D-glucose showed a transient overshoot in the presence of Na+ gradient between medium and vesicles (medium greater than vesicles). The overshoot magnitude was increased (1.8-fold of controls) in diabetic brush border membrane vesicles and recovered to the control level by the treatment of diabetic rats with insulin. In contrast, increased uptake of D-glucose in diabetic rats was not recovered by the starvation of diabetic rats although the blood glucose level was the same as that of controls. Furthermore, we attempted to examine phlorizin binding activities among four groups. Scatchard analysis indicated that phlorizin binding to diabetic brush border membrane vesicles was increased (1.6-fold of controls) without a change of the affinity for phlorizin as compared with controls. Increased binding of phlorizin to diabetic brush border membrane vesicles was also recovered to the control level by the treatment of diabetic rats with insulin, but not by starvation. These results suggested that the increased activity of Na+/glucose cotransporter in diabetic rats was due to the increase of the number of cotransporter and that intestinal cotransporter was physiologically controlled by insulin, but not by blood glucose levels.     

Determination of dissolved and particulate thiols in Lake Biwa water and extracted fulvic acids by solid phase extraction followed by HPLC with fluorescence detection

Thiols are important antioxidants that can modulate the bioavailability and biogeochemistry of many soft metals, although their detection remains challenging in both their reduced (R–S) and oxidized (R–S–S–R) forms. Here, a modified biochemical method was applied to determine the levels of dissolved and particulate thiols in Lake Biwa water and extracted Lake Biwa fulvic acids obtained at various depths. This method involves the use of the reducing agent tris(2-carboxyethyl)phosphine and the fluorescent label 7-fluorobenzofurazan-4-sulfonic acid ammonium salt (SBD-F), followed by solid-phase extraction and HPLC with fluorescence detection. Dissolved cysteine (Cys) (2.0–6.0 nM), glutathione (GSH) (2.8–5.1 nM), and N-acetyl-l-cysteine (NAC) (1.6–4.2 nM) were detected throughout the water column but were broadly consistent at depths of 5–20 m. In contrast, abundant levels of particulate cysteine (1.3–3.5?×?10² nM) and glutathione (1.6–3.1?×?10² nM) were detected down to depths of 15 m. The particulate cysteine and glutathione were significantly covariant, and the ratios between them reflected the differences in the plankton community composition and availability of these compounds. This work also studied the concentrations of Cys, GSH and NAC in Lake Biwa fulvic acids (LBFAs) for the first time (at 0 m: cysteine, 0.8 nM; glutathione, 1.6 nM; NAC, 2.5 nM; at 10 m: cysteine, 1.4 nM; glutathione, 0.6 nM; NAC, 1.6 nM). The nanomolar to sub-nanomolar concentrations of the particulate and dissolved Cys, GSH and NAC in the lake indicates that these are an important class of ligands for chalcophile metals and may influence the distribution of plankton communities from the epilimnion to the hypolimnion of the lake.