Comparative Proteomics on Deep-Sea Amphipods after in Situ Copper Exposure.

The interest in deep-sea mining increased along with the environmental concerns of these activities to the deep-sea fauna. The discovery of optimal biomarkers of deep-sea mining activities in deep-sea species is a crucial step toward the supply of important ecological information for environmental impact assessment. In this study, an in situ copper exposure experiment was performed on deep-sea scavenging amphipods. Abyssorchomene distinctus individuals were selected among all the exposed amphipods for molecular characterization. Copper concentration within the gut was assessed, followed by a tandem mass tag-based coupled with two-dimensional liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) applied to identify and quantify the protein expression changes after 48 h of exposure. 2937 proteins were identified and annotated, and 1918 proteins among all identified proteins were assigned by at least two nonambiguous peptides. The screening process was performed based on the differences in protein abundance and the specific correlation between the proteins and copper in previous studies. These differentially produced proteins include Na+/K+ ATPase, cuticle, chitinase, and proteins with unknown function. Their abundances showed correlation with copper and had high sensitivity to indicate the copper level, being here proposed as biomarker candidates for deep-sea mining activities in the future. This is a key step in the development of environmental impact assessment of deep-sea mining activities integrating ecotoxicological data.

Complete mitochondrial genome of Hydrolagus mitsukurii (Jordan & Snyder, 1904).

Holocephali has foreseeable value to help our understanding of vertebrate genome evolution due to its phylogenetic position. In this study, we reported a complete mitochondrial genome of Hydrolagus mitsukurii, a species of holocephalans. The mitochondrial genome is 20,486 bp in length and comprised 13 PCGs, 2 rRNA, 22 tRNA, and 1 control region (D-loop), as well as a long noncoding insertion between tRNAThr and tRNAPro. A phylogenetic tree based on 13 PCGs showed that Hydrolagus mitsukurii was grouped with the members of the family Chimaeridae. Furthermore, the phylogenetic tree further supported the paraphyletic clades of Hydrolagus and Chimera.

First record of a deep-sea tardigrade from the South China Sea, Halechiniscus janus sp. nov. (Arthrotardigrada: Halechiniscidae).

The knowledge of marine tardigrades in the South China Sea (Western Pacific Ocean) is very scarce, with only four species from shallow waters recorded until now. The first deep-sea (15301624 m bsl) tardigrade species from this sea, Halechiniscus janus sp. nov. (Arthrotardigrada: Halechiniscidae), is described here. Specimens of the new species have four wrinkled digits without peduncles on each leg, terminated by simple crescent-shaped claws, and primary clavae clearly shorter than cirri A, both inserted on a common cirrophore, as typical for the genus Halechiniscus Richters, 1908. The new species differs from all other Halechiniscus species by its cylindrical body and conical head; by the presence of semispherical secondary clavae, and by sensory organs on legs IV consisting of a short cirrophore followed by a short papilla terminated in a peculiar short bipartite tip. The discovery of this new bathyal species, with its peculiar morphological traits, brings new insights not only to the biogeography and ecology of tardigrades, but also to the understanding of the only partially resolved systematics of the diversified family Halechiniscidae.

The complete mitogenome of the blackgill catshark, Parmaturus melanobranchus (Chan, 1966).

A rare specimen of Parmaturus melanobranchus was collected from the South China Sea. The complete mitochondrial genome of the specimen was sequenced using Illumina Hiseq platform and assembled with Geneious and Trinity. The mitogenome is 16,687 bp long with a base composition of 30.4% A, 14.1% G, 23.5% C and 32.1% T, respectively. A total of 37 genes were predicted containing 13 protein-coding genes, 2 ribosomal RNA genes and 22 transfer RNA genes. This is the first complete mitochondrial genome published of this species.

Carbonic anhydrase II regulates differentiation of ameloblasts via intracellular pH-dependent JNK signaling pathway.

Differentiation of ameloblasts from undifferentiated epithelial cells is controlled by diverse growth factors, as well as interactions between epithelium and mesenchyme. However, there is a considerable lack of knowledge regarding the precise mechanisms that control ameloblast differentiation and enamel biomineralization. We found that the expression level of carbonic anhydrase II (CAII) is strongly up-regulated in parallel with differentiation of enamel epithelium tissues, while the enzyme activity of CA was also increased along with differentiation in ameloblast primary cultures. The expression level of amelogenin, a marker of secretory-stage ameloblasts, was enhanced by ethoxzolamide (EZA), a CA inhibitor, as well as CAII antisense (CAIIAS), whereas the expression of enamel matrix serine proteinase-1 (EMSP-1), a marker for maturation-stage ameloblasts, was suppressed by both. These agents also promoted ameloblast proliferation. In addition, inhibition of ameloblast differentiation by EZA and CAIIAS was confirmed using tooth germ organ cultures. Furthermore, EZA and CAIIAS elevated intracellular pH in ameloblasts, while experimental decreases in intracellular pH abolished the effect of CAIIAS on ameloblasts and triggered the activation of c-Jun N-terminal kinase (JNK). SP600125, a JNK inhibitor, abrogated the response of ameloblasts to an experimental decrease in intracellular pH, while the inhibition of JNK also impaired ameloblast differentiation. These results suggest a novel role for CAII during amelogenesis, that is, controlling the differentiation of ameloblasts. Regulation of intracellular pH, followed by activation of the JNK signaling pathway, may be responsible for the effects of CAII on ameloblasts.

Effect of glypican-1 gene on the pulp cells during the reparative dentine process.

GPC-1 (glypican-1) is a cell surface heparan sulfate proteoglycan that acts as a co-receptor for heparin-binding growth factors and members of the TGF-ß (transforming growth factor beta-1) family. The function of cell-surface proteoglycans in the reparative dentine process has been under investigation. Gpc-1 was detected with similar frequency as tgf-ß1 in the cDNA library using mRNA from the odontoblast-like cell-enriched pulp of rat incisors. The aim of this study was to test our hypothesis that gpc-1 may be related to reparative dentine formation. We examined the expression of this gene during the reparative dentine process, as well as the effect of gpc-1 on odontoblast-like cell differentiation using siRNA (small interfering RNA) to down-regulate gpc-1 expression. Immunohistological examination showed that GPC-1 was expressed in pulp cells entrapped by fibrodentine and odontoblast-like cells as well as TGF-ß1. The mRNAs for gpc-1, -3 and -4, except for gpc-2, were expressed during odontoblast-like cell differentiation in pulp cells. The relative levels of gpc-1 mRNA were increased prior to the differentiation stages and were decreased during the secretory and maturation stages of pulp cells. Down-regulation of gpc-1 expression resulted in a 3.9-fold increase in tgf-ß1 expression in pulp cells and a 0.3-fold decrease in dspp (dentine sialophosphoprotein) expression compared with control. These results suggested that gpc-1 and tgfß-1 expression are necessary for the onset of differentiation, but should be down-regulated before other molecules are implicated in the formation of reparative dentine. In conclusion, gpc-1 expression in odontoblast-like cells is associated with the early differentiation but not with the formation of reparative dentine.

The mitochondrial genome of a deep-sea oplophoroid of the genus Acanthephyra (Crustacea: Caridea: Oplophoroidea).

The genus Acanthephyra mainly inhabits deep waters with the maximum depth exceeding 5000 m. It has a wide distribution, except in high latitude areas. Here, we report the mitochondrial genome of Acanthephyra sp. which was collected from the northeast of South China Sea. The genome is 16,205 bp in length with a 61.52% AT content. It contains 13 protein-coding genes (PCGs), 2 ribosomal RNA genes, and 22 transfer RNA genes. Phylogenetic analysis shows that the present species is closest to A. smithi and Oplophoroidea has a close relationship with Bresilioidea in Caridea.

Moebjergarctus clarionclippertonensis, a new abyssal tardigrade (Arthrotardigrada, Halechiniscidae, Euclavarctinae) from the Clarion-Clipperton Fracture Zone, North-East Pacific.

A new marine tardigrade, Moebjergarctus clarionclippertonensis sp. nov., is described based on specimens collected from a manganese nodule area in the Clarion-Clipperton Fracture Zone of the abyssal North-eastern Pacific. The new species is a member of the bathyal/abyssal subfamily Euclavarctinae Renaud-Mornant, 1983. Within the Euclavarctinae, the genus Moebjergarctus Bussau, 1992, with only one described species, M. manganis Bussau, 1992, is characterised by simple claws, club-shaped and anteriorly bent primary clavae, well-developed spherical secondary clavae and cephalic cirri separated into three parts: short cirrophore, long and annulated scapus, and a short flagellum. Moebjergarctus clarionclippertonensis sp. nov. can be distinguished from M. manganis by the morphology of cephalic cirri which have scapi annulated only in the proximal part and by the presence of a caudodorsal bulge covered by a crescent-shaped cuticular thickening.

Characterization of synovial cell clones isolated from rheumatoid arthritis patients: possible involvement of TNF-alpha in reduction of osteoprotegerin in synovium.

To elucidate the role of the synovium in bone destruction by osteoclasts in rheumatoid arthritis (RA), primary synovial cells isolated from RA patients were cultured and characterized. The cultured primary cells did not produce RANKL (TRANCE/ODF/OPGL/TNFSF11/CD254), an inducer of osteoclast differentiation, but constitutively produced its inhibitor, osteoprotegerin (OPG). Addition of TNF-alpha to the primary cultures of synovial cells reduced the cell viability and strongly suppressed OPG production. We then established nine synovial cell clones, including SYM-1, responsible for OPG production from primary synovial cell cultures. TNF-alpha induced apoptosis of SYM-1 cells within 24h and decreased OPG levels, while infliximab, a chimerical form of the anti-TNF-alpha antibody drug, suppressed the apoptosis and restored OPG levels. These results suggest the existence of fibroblastic cells producing OPG in the synovium, while TNF-alpha suppresses OPG production by inducing apoptosis in those cells. Further, infliximab is considered to inhibit bone destruction through restoration of OPG levels in RA.

The differences in epidemiological and psychological features of globus symptoms between urban and rural Guangzhou, China: A cross-sectional study.

To compare the epidemiological and psychological features of globus symptoms between individuals from urban and rural areas in Guangzhou.In total, 3360 individuals aged 18 years and over were selected to participate in our questionnaire investigation using random cluster sampling under the stratification of a urban area and a rural area. The questionnaire comprised questions on personal characteristics and globus symptomatology and psychological characteristic and sleep quality scales.Lifetime prevalence and Glasgow-Edinburgh throat scale scores of globus symptoms were greater in the urban area than in the rural area, but no significant differences in sex ratio or onset age between individuals with globus were found. The incidences and severity of anxiety, depression, and sleep disorders were significantly higher among patients who presented with globus in the urban area than among those in the rural area.The lifetime prevalence of globus symptoms and the psychological features of globus patients differ between urban and rural inhabitants. We should pay more attention to these differences.

Effect of a novel Er:YAG laser in caries removal and cavity preparation: a clinical observation.

OBJECTIVE: We evaluated the applicability of a novel Er:YAG laser under clinical conditions. BACKGROUND DATA: The Er:YAG laser has been demonstrated to be a safe and effective alternative to the conventional turbine bur, but the relatively low cutting speed prevented the wide application of Er:YAG laser in clinical cavity preparation. METHODS: A Smart 2940 D laser developed by Deka Corporation was used for cavity preparation in 95 teeth of 45 patients. Parameters were as follows: wavelength 2.94 microm, pulse energy 700 mJ, repetition rate 8 Hz. Pain, discomfort, assessment during cavity preparation, prognosis factor, and overall clinical evaluation were assessed during or after treatment. RESULTS: No adverse reaction was observed in any tooth. No intraoperative pain or only slight intraoperative pain was described in 85 teeth (89.5%). Cavity preparation was completed with the laser system alone in 90 teeth (94.7%). Overall clinical evaluation showed no safety problems, with a very good or good rating in 86 teeth (90.5%). The overall operation time was 49 sec on average. CONCLUSION: The Smart 2940 D is an efficient, effective, safe, and suitable instrument for caries removal and for cavity preparation. It greatly shortens operation time.

Effects of KTP laser irradiation, diode laser, and LED on tooth bleaching: a comparative study.

OBJECTIVE: This in vitro study examines the whitening efficacy of a light-emitting diode (LED), a diode laser, and a KTP laser irradiation in dental bleaching by analyzing the change in color achieved from the treatment, the temperature increase induced in the pulpal cavity, as well as enamel microhardness measurement after treatment. BACKGROUND DATA: Bleaching techniques achieved significant advances with the use of coherent or incoherent radiation sources to activate the bleaching agents. METHODS: A hydrogen peroxide bleaching agent, Hi-Lite, was stimulated with an LED, a 980-nm diode laser at 0.8 W, or a 532-nm KTP laser at 1.0 W for 30 sec on 64 extracted human incisors. During irradiation, the temperature in the pulpal cavity was monitored. The color change was evaluated using the CIE L*a*b* color space measurement system, and Vikers enamel microhardness was tested after treatment. RESULTS: A mean total color difference value (DeltaE*) greater than 5.0 was obtained in each group. KTP-laser-induced bleaching gave a significantly higher DeltaL* (8.35) after treatment (p < 0.01). Neither LED nor the two lasers produced significant differences in the enamel microhardness after treatment (p > 0.01). Mean maximal pulpal temperature rise was 2.95 degrees C for LED, 3.76 degrees C for KTP laser, and 7.72 degrees C for diode laser, respectively. CONCLUSION: The results of this study suggest that KTP laser is effective at providing brighter teeth. According to the conditions used in this study, the LED and KTP laser induced a safer pulpal temperature increase when assisted with Hi-Lite bleaching gel.

Healing process of skin after CO2 laser ablation at low irradiance: a comparison of continuous-wave and pulsed mode.

OBJECTIVE: The purpose of the present study was to compare the healing process of rabbit's skin after the ablation of continuous wave CO(2) laser and pulsed CO(2) laser at low irradiances. BACKGROUND DATA: Because of its advantages, the CO(2) laser has been viewed as an alternative to the traditional scalpel. Simple thermal models suggest that suitably short pulses of laser irradiation can leave a smaller thermal damaged zone to ablated tissue, which may lead to faster healing. METHODS: One continuous-wave (cw, 2 watt) and two pulsed CO(2) lasers (sp(1), 100 Hz, 2 watt; and sp(2), 25 Hz, 2 watt) were used to ablate twelve rabbits' ear skin for 2 min, respectively. After sacrifice at 0, 3, 7, 14, 28, and 56 days postsurgery, gross observation and histological examination were performed. RESULTS: sp(1) and sp(2) resulted in similar ablation rates to cw laser. At 3, 7, 14, and 28 days postsurgery, the scores of the group cw were significantly lower than both the sp(1) and sp(2) scores (p < 0.01). From 7 to 28 days, wounds of sp1 showed the best histological outcomes. CONCLUSION: At a relatively low irradiance, pulsed CO(2) laser is capable of bloodless skin ablation with improved wound healing. Increasing repetition rate of the CO(2) laser may help to achieve a better surgical outcome. Pulsed CO(2) laser may be a valuable instrument for ablation skin and skin lesions.

Effects of diode laser irradiation on smear layer removal from root canal walls and apical leakage after obturation.

OBJECTIVE: The objective of this study was to investigate the rise in temperature in root surfaces during and immediately after diode laser irradiation, to observe morphological changes of root canal wall after irradiation, and to evaluate the apical leakage after irradiation and obturation in vitro. BACKGROUND DATA: There have been very few reports on root canal treatment by 980-nm wavelength diode laser. METHODS: Sixty-six extracted human single-rooted teeth were instrumented up to size 60 K-file, and then randomly divided into three groups of 22 teeth each. Groups 1 and 2 were irradiated with a diode laser at 5 W for 7 sec using fibers of diameters 550 and 365 microm, respectively. Group 3 was not irradiated, and served as a control. The rise in temperature on root surfaces of the teeth in groups 1 and 2 were measured by thermography. Six teeth in each group were bisected longitudinally and observed morphologically. Other teeth were obturated and immersed in rhodamine B solution, and the degree of apical leakage was evaluated longitudinally and transversally. RESULTS: A maximum temperature rise of 8.1( degrees )C was recorded in group 1. The smear layer in the laser-treated groups was evaporated and removed, resulting in clean root canal walls, which was significantly superior to the control group (p < 0.05). After obturation, the laser-treated groups showed significantly less apical leakage than the control group (p < 0.05). CONCLUSIONS: These results indicate that the diode laser is useful for removing smear layer and debris from root canal walls, and reducing apical leakage after obturation in vitro, and suggest that it would be useful for root canal treatment in clinic.

Synthetic octacalcium phosphate-enhanced reparative dentine formation via induction of odontoblast differentiation.

Synthetic octacalcium phosphate (OCP) has been suggested to be a useful biomaterial for the regeneration of hard tissues, including bone. However, it remains unknown whether OCP induces dentine formation by dental pulp. We investigated biomineralization of dental pulp exposed to synthetic OCP in vitro and in vivo. When dental pulp was exposed directly to OCP, rapid formation of reparative dentine (RD) was induced and expression of dentine sialoprotein synthesis was observed in dental pulp adjacent to newly synthesized RD. OCP inhibited the proliferation of rat pulp cells and also promoted their odontoblastic differentiation in vitro, as alkaline phosphatase activity, mineralization of pulp cells and the expression level of dentine sialophosphoprotein were enhanced. Direct contact between OCP and pulp cells is required for OCP to exhibit its effects in vitro. The expression level of Runx2, a transcription factor whose downregulation is closely related to odontoblast differentiation, was downregulated in pulp cells cultured with OCP. Structural changes of OCP during culture were determined by Fourier transform infrared spectroscopy. OCP tended to be converted to carbonate hydroxyapatite after incubation with or without pulp cells, which may be analogous to biological apatite crystals. Taken together, our data suggest that synthetic OCP supports RD formation by dental pulp and downregulation of Runx2 may be involved in that stimulatory activity. Furthermore, OCP-apatite conversion is involved in this stimulatory capacity of OCP.

Histopathological study of dental pulp tissue capped with enamel matrix derivative.

The purpose of this study was to examine the histopathological response of dental pulp tissue to enamel matrix derivative (EMD) used as a pulp capping material. Thirty-two teeth from two mongrel dogs were divided into four equal groups. One group served as controls, and the others were used for deep Class V cavity preparation followed by direct pulp capping with enamel matrix derivative. The treated teeth were extracted after 1, 4, and 8 weeks and prepared for histopathological examination by light microscopy. All teeth prepared after 4 and 8 weeks demonstrated an increase in tertiary dentin, suggesting that enamel matrix derivative exerts a considerable influence on odontoblasts and endothelial cells of capillaries in dental pulp tissue. These results imply that enamel matrix derivative used as a pulp capping material may play a role in the calcification of dental pulp tissue.

Thermographical and morphological studies of Er,Cr:YSGG laser irradiation on root canal walls.

OBJECTIVE: This study was performed to investigate the temperature rises on root surfaces and morphological changes on root canal walls of extracted human teeth after irradiation by erbium, chromium: yttrium, scandium, gallium, garnet (Er, Cr:YSGG) laser and to evaluate the efficiency in removing smear layer and debris from the prepared root canal walls in vitro. BACKGROUND DATA: No report on Er, Cr:YSGG laser effects under various conditions and specific tips for endodontic treatments has been reported. METHODS: Sixty extracted human teeth with single and straight root were used in this study. The teeth were prepared at 1 mm short of the apical foramen by a conventional technique using K-files. Fifty-four specimens were irradiated by Er, Cr:YSGG laser at the output powers of 2 W, 3 W, and 5 W for 7 sec with three fiber tips (200, 320, and 400 microm diameters). Temperature changes were recorded thermographically. All teeth were examined by light microscopy or scanning electron microscopy (SEM). RESULTS: Thermographic study showed that the average temperature rises on root surfaces were less than 8 degrees C. Observation by light microscopy revealed the ablation at the apical stop, and that by SEM indicated that laser irradiation at 5 W using a fiber tip with 400 microm diameter was efficient for removing smear layer and debris without carbonization or melting. CONCLUSIONS: The results of this study suggested that the temperature rises during Er, Cr:YSGG laser irradiation at the parameters used in this study are minimal to cause the damage on periodontal and bone tissues. Moreover, it was suggested that it is efficient to remove smear layer and debris without causing any carbonization and melting.

Pseudoalteromonas lipolytica sp. nov., isolated from the Yangtze River estuary.

A strictly aerobic, Gram-negative, non-pigmented bacterial strain, designated LMEB 39(T), was isolated from a seawater sample collected from the Yangtze River estuary near the East China Sea and was examined physiologically, chemotaxonomically and phylogenetically. The novel isolate was motile by a single polar flagellum and positive for nitrate reduction and decomposition of casein, gelatin, Tween 20 and Tween 80, but negative for indole production. Chemotaxonomic analysis revealed ubiquinone-8 as the predominant respiratory quinone and phosphatidylethanolamine and phosphatidylglycerol as major polar lipids. The major fatty acids were C(16 : 1) ω 7c/iso-C(15 : 0) 2-OH, C(16 : 0), C(18 : 1) ω 7c, C(12 : 0) 3-OH, C(17 : 1)ω 8c and C(17 : 0). The genomic DNA G+C content was 42.3 mol%. Comparative 16S rRNA gene sequence analysis revealed that the isolate belongs to the genus Pseudoalteromonas. Strain LMEB 39(T) exhibited the closest phylogenetic affinity to Pseudoalteromonas byunsanensis JCM 12483(T) (97.4 % sequence similarity). The DNA-DNA reassociation values between strain LMEB 39(T) and P. byunsanensis JCM 12483(T) and Pseudoalteromonas undina DSM 6065(T) (97.2 % sequence similarity) were 31.7 and 30.3 %, respectively. On the basis of phenotypic and genotypic data, strain LMEB 39(T) represents a novel species of the genus Pseudoalteromonas, for which the name Pseudoalteromonas lipolytica sp. nov. is proposed; the type strain is LMEB 39(T) (=CGMCC 1.8499(T)=JCM 15903(T)).

Effects of mineral trioxide aggregate on the differentiation of rat dental pulp cells.

The effect of mineral trioxide aggregate (MTA) on the odontoblast-like differentiation of pulp cells was evaluated using heat-shock protein 25 (hsp25) as a marker for odontoblast differentiation. The cells were cultured with tooth-colored MTA or calcium hydroxide-containing cement (Dycal). The effects of the materials on the pulp cells were observed using a confocal laser scanning microscope. The cells were labelled immunocytochemically using polyclonal antibodies against hsp25 and actin. The mRNA expression of hsp25 and dspp in the pulp cells at 2 days were examined by quantitative reverse transcription-polymerase chain reaction (RT-PCR). Most of the cells cultured with MTA showed an intense immunolabelling for hsp25 and the mRNA expressions of hsp25 and dspp at 2 days were higher than those cultured with Dycal. These findings indicate that MTA is an effective pulp capping material and is able to induce the differentiation of odontoblast-like cells and the formation of reparative tertiary dentin with minimum apoptosis.

Croceicoccus marinus gen. nov., sp. nov., a yellow-pigmented bacterium from deep-sea sediment, and emended description of the family Erythrobacteraceae.

A Gram-negative, aerobic, neutrophilic, coccoid bacterium, strain E4A9T, was isolated from a deep-sea sediment sample collected from the East Pacific polymetallic nodule region. 16S rRNA gene sequence analysis showed that the isolate was related to the type strain of Altererythrobacter epoxidivorans (96.0% sequence similarity). Lower 16S rRNA gene sequence similarities were observed with other members of the genera Altererythrobacter (94.7%), Erythrobacter (94.0-95.4%), Erythromicrobium (94.8%) and Porphyrobacter (94.6-95.1%) of the family Erythrobacteraceae. Phylogenetic analysis including all described species of the family Erythrobacteraceae and several members of the family Sphingomonadaceae revealed that the isolate formed a distinct phylogenetic lineage with the family Erythrobacteraceae. Chemotaxonomic analysis revealed ubiquinone-10 as the predominant respiratory quinone, anteiso-C15:0, iso-C14:0 and iso-C15:0 as major fatty acids, and phosphatidylglycerol as the major polar lipid. The DNA G+C content was 71.5 mol%. The isolate contained carotenoids, but no bacteriochlorophyll a. On the basis of phenotypic and genotypic data presented in this study, strain E4A9T represents a novel species in a new genus in the family Erythrobacteraceae for which the name Croceicoccus marinus gen. nov., sp. nov. is proposed; the type strain is E4A9T (=CGMCC 1.6776T=JCM 14846T).