Transgenerational acclimation to acidified seawater and gene expression patterns in a sea urchin.

Transgenerational responses of susceptible calcifying organisms to progressive ocean acidification are an important issue in reducing uncertainty of future predictions. In this study, a two-generation rearing experiment was conducted using mature Mesocentrotus nudus, a major edible sea urchin that occurs along the coasts of northern Japan. Morphological observations and comprehensive gene expression analysis (RNA-seq) of resulting larvae were performed to examine transgenerational acclimation to acidified seawater. Two generations of rearing experiments showed that larvae derived from parents acclimated to acidified seawater tended to have higher survival and show less reduction in body size when exposed to acidified seawater of the same pH, suggesting that a positive carry-over effect occurred. RNA-seq analysis showed that gene expression patterns of larvae originated from both acclimated and non-acclimated parents to acidified seawater tended to be different than control condition, and the gene expression pattern of larvae originated from acclimated parents was substantially different than that of larvae of non-acclimated and control parents.

Whole transcriptome analysis of demersal fish eggs reveals complex responses to ocean deoxygenation and acidification.

Ocean acidification and deoxygenation co-occur in marine environments, causing deterioration of marine ecosystems. However, effects of compound stresses on marine organisms and their physiological coping mechanisms are largely unknown. Here, we show how high pCO2 and low dissolved oxygen (DO) cause transcriptomic changes in eggs of a demersal fish (Sillago japonica), which are fully exposed to such stresses in natural environment. Overall gene expression was affected more strongly by low DO than by high pCO2. Enrichment analysis detected significant stress responses such as glycolytic processes in response to low DO. Increased expression of a group of glycolytic genes under low DO conditions is presumably because oxygen depletion disables the electron transfer pathway, complementing ATP production in the glycolytic pathway. Contrary to expectations, apparent mitigation of gene expression changes was dominant under combined stress conditions, and may represent an innate fish adaptive trait for severe environments.

Interactive effects of ocean deoxygenation and acidification on a coastal fish Sillago japonica in early life stages.

Acidification and deoxygenation are major threats to ocean environments. Despite the possibilities of their co-occurrence, little is known about their interactive effects on marine organisms. The effects of low pH and low dissolved oxygen (DO) on the early life stages of the coastal fish Sillago japonica were investigated. Twenty-five experimental treatments fully crossed in five levels of pH 7.6-8.1 and DO 50-230 µmol/kg (20-100 % saturation degree) were tested, and hatching rate of the embryos and survivability of the larvae after 24 h at 25 °C were investigated. Low DO treatment significantly affected the embryos and larvae compared to low pH treatment. The 50 % lethal concentration of DO showed the highest value at pH 7.6 and the lowest value at pH 7.7 and 7.9 for embryos and larvae, respectively. Therefore, effects of deoxygenation on fishes were alleviated under acidified condition around pH 7.7-7.9.

Orphan gene expressed in flame cone cells uniquely found in seahorse epithelium.

The seahorse is one of the most unique teleost fishes in its morphology. The body is surrounded by bony plates and spines, and the male fish possess a brooding organ, called the brood pouch, on their tail. The surfaces of the brood pouch and the spines are surrounded by characteristic so-called flame cone cells. Based on our histological observations, flame cone cells are present in the seahorse Hippocampus abdominalis, but not in the barbed pipefish Urocampus nanus or the seaweed pipefish Syngnathus schlegeli, both of which belong to the same family as the seahorse. In the flame cone cells, we observed expression of an "orphan gene" lacking homologs in other lineages. This gene, which we named the proline-glycine rich (pgrich) gene, codes for an amino acid sequence composed of repetitive units. In situ hybridization and immunohistochemical analyses detected pgrich-positive signals from the flame cone cells. Based on a survey of the genome sequences of 15 teleost species, the pgrich gene is only found from some species of Syngnathiformes (namely, the genera Syngnathus and Hippocampus). The amino acid sequence of the seahorse PGrich is somewhat similar to the sequence deduced from the antisense strand of elastin. Furthermore, there are many transposable elements around the pgrich gene. These results suggest that the pgrich gene may have originated from the elastin gene with the involvement of transposable elements and obtained its novel function in the flame cone cells during the evolution of the seahorse.

Brood pouch evolution in pipefish and seahorse based on histological observation.

INTRODUCTION: Fishes of the Syngnathidae family are rare in having male pregnancy: males receive eggs from females and egg development occurs in the male brood pouch that diverged during evolution. The family is divided into two subfamilies: Nerophinae and Syngnathinae. METHODS: We compared histologically five types of the brood pouch in Syngnathinae: an open pouch without skinfolds (alligator pipefish); an open pouch with skinfolds (messmate pipefish); a closed pouch with skinfolds (seaweed pipefish); and closed pouches with a sac-like pouch on the tail (pot-bellied seahorse) or within a body cavity (Japanese pygmy seahorse). RESULTS: Histological observations revealed that all the examined species possess vascular egg compartments during the brooding period. The present immunohistochemical study revealed that the pregnant egg compartment epithelium grows thin in both open and closed pouches. The placenta of open and closed pouches is composed of dermis and reticulin fibers, respectively. The closed pouch placenta is a flexible and moist tissue, suitable for substance transport between the father and embryos through the epithelium and blood vessels and responsible for supplying nutrition and removing waste. DISCUSSION: These results suggest that the basic egg incubation structures were established at an early stage of Syngnathinae evolution. On the other hand, it is likely that the innovation of tissue structure, where dermis was replaced with reticular fibers, occurred in closed brood pouches to regulate the pregnant pouch environment. The present study presents the morphological evolutionary pathway of the brood pouch in Syngnathinae, providing a basis for further molecular-level evolutionary studies.

Unique environmental Symbiodiniaceae diversity at an isolated island in the northwestern Pacific.

Dinoflagellates in the family Symbiodiniaceae are intensively investigated as algal symbionts of corals and other invertebrates, underpinning coral reef ecosystems as primary producers. Diversity, including regional diversification, of free-living communities is less studied. In this study, an environmental Symbiodiniaceae community at an isolated island, Okinotori Island, Japan, was investigated to determine whether the community is endemic or common with other locations near continents and major ocean currents. Symbiotic algae in common corals at the island were the same type as those of the corals from other Japanese waters. In the environmental samples, genera Symbiodinium (formerly clade A), Cladocopium (clade C), Durusdinium (clade D), and clades F (including Freudenthalidium), G, and I, were identified through analysis of internal transcribed spacer region 2 of nuclear ribosomal RNA gene (ITS2) sequences. Interestingly, some sequences found were genetically different from those of previously reported genera/clades. These unknown sequences were genetically included in the Symbiodiniaceae linage, but they were differentiated from the previously known nine clades. The sequences formed a cluster in the phylogenetic tree based on 28S nrDNA. These sequences were thus considered members of a novel clade in the family (clade J). In total, 120 kinds of ITS2 sequences were produced; while 10 were identical to previously reported sequences, the majority were highly divergent. These genetically unique Symbiodiniaceae types, including novel clade J, may have evolved in isolation and reflect the environmental characteristics of the Okinotori Island.

Wear Simulation of Ceramic-on-Crosslinked Polyethylene Hip Prostheses: A New Non-Oxide Silicon Nitride versus the Gold Standard Composite Oxide Ceramic Femoral Heads.

The purpose of the present study was to compare the wear behavior of ceramic-on-vitamin-E-diffused crosslinked polyethylene (Vit-E XLPE) hip bearings employing the gold standard oxide ceramic, zirconia (ZrO2)-toughened alumina (Al2O3) (ZTA, BIOLOX®delta) and a new non-oxide ceramic, silicon nitride (Si3N4, MC2®). In vitro wear test was performed using a 12-station hip joint simulator. The test was carried out by applying the kinematic inputs and outputs as recommended by ISO 14242-1:2012. Vitamin-E-diffused crosslinked polyethylene (Vit-E XLPE) acetabular liners (E1®) were coupled with Ø28-mm ZTA and Si3N4 femoral heads. XLPE liner weight loss over 5 million cycles (Mc) of testing was compared between the two different bearing couples. Surface topography, phase contents, and residual stresses were analyzed by contact profilometer and Raman microspectroscopy. Vit-E XLPE liners coupled with Si3N4 heads produced slightly lower wear rates than identical liners with ZTA heads. The mean wear rates (corrected for fluid absorption) of liners coupled with ZTA and Si3N4 heads were 0.53 ± 0.24 and 0.49 ± 0.23 mg/Mc after 5 Mc of simulated gait, respectively. However, after wear testing, the ZTA heads retained a smoother topography and showed fewer surface stresses than the Si3N4 ones. Note that no statistically significant differences were found in the above comparisons. This study suggests that the tribochemically formed soft silica layer on the Si3N4 heads may have reduced friction and slightly lowered the wear of the Vit-E XLPE liners. Considering also that the toughness of Si3N4 is superior to ZTA, the present wear data represent positive news in the future development of long-lasting hip components.

Highly Porous Titanium Cups versus Hydroxyapatite-Coated Sockets: Midterm Results in Metachronous Bilateral Total Hip Arthroplasty.

OBJECTIVE: Highly porous titanium cups have been recently introduced, with contradictory outcomes. A retrospective consecutive case series involving bilateral metachronous total hip arthroplasties (THA) performed with 2 different cups, i.e., Fixa (F) and Fixa Ti-Por (T) (Adler Ortho, Milan, Italy), and the same stem, was evaluated. T sockets, manufactured using electron beam melting, were supposed to prove superior in terms of clinical results, survival rates, and radiographic parameters in comparison to hydroxyapatite-coated F cups with conventional porosity. SUBJECTS AND METHODS: Twenty-four bilateral metachronous THAs with an F cup on one side and a T socket on the other side were evaluated. Preoperative and postoperative Harris hip scores (HHS) were collected for every patient. Radiographic signs of loosening were assessed. The radiographic signs of osseointegration (radiolucent lines, superolateral buttress, inferomedial buttress, radial trabeculae, and stress shielding) were evaluated. RESULTS: No intraoperative complications occurred. The mean HHS score was excellent and comparable in both groups. At the mean follow-up of 134 months (F) and 79 months (T), no cup or liner revisions were performed. No radiographic signs of loosening were reported. All of the patients revealed 3 parameters of good bony ingrowth at least. Both groups showed similar radiographic parameters regarding osseointegration, which were stable over the time. Stress shielding was more evident in the T cohort (p =0.07). CONCLUSION: Highly porous titanium cups produced using an additive manufacturing and electron beam melting technology achieved reliable midterm clinical and radiographic results not inferior to those of second-generation cups.

Shift of symbiont communities in Acropora tenuis juveniles under heat stress.

Ocean warming is a major threat to coral reefs, leading to an increasing frequency and amplitude of coral bleaching events, where the coral and its algal symbiont associations breakdown. Long-term change and resilience of a symbiont community in coral juveniles is thought to be one of the most important aspects for determining thermal tolerance of the coral holobionts; however, despite its importance, they are not well documented in both under elevated temperature and even under natural condition. Here we investigated changes in symbiont communities in juveniles of the coral Acropora tenuis under controlled heat stress conditions (30 °C, 31/32 °C) and natural variations in seawater temperatures (19-30 °C) for up to four months. Compared with the ambient temperature conditions, coral survival rates were higher when exposed to 30 °C, but survival rates decreased when exposed to 31/32 °C. Symbiodinium types A3, C1, and D1-4 were detected in the juveniles under all thermal conditions; however, in higher water temperatures (31/32 °C), both the prevalence of D1-4 Symbiodinium and the number of juveniles harboring only this type of symbiont increased after two to four months later. In contrast, colonies at lower temperatures (30 °C and ambient) harbored multiple clades of symbionts over the same experimental period. These results highlight the flexibility of the coral-Symbiodinium symbiosis for juvenile A. tenuis under variable thermal conditions. In particular, the benefit of the preferential association with type D1-4 can be considered as a response when under heat-stress conditions, and that could help corals to cope with ocean warming.

In Vitro versus In Vivo Phase Instability of Zirconia-Toughened Alumina Femoral Heads: A Critical Comparative Assessment.

A clear discrepancy between predicted in vitro and actual in vivo surface phase stability of BIOLOX®delta zirconia-toughened alumina (ZTA) femoral heads has been demonstrated by several independent research groups. Data from retrievals challenge the validity of the standard method currently utilized in evaluating surface stability and raise a series of important questions: (1) Why do in vitro hydrothermal aging treatments conspicuously fail to model actual results from the in vivo environment? (2) What is the preponderant microscopic phenomenon triggering the accelerated transformation in vivo? (3) Ultimately, what revisions of the current in vitro standard are needed in order to obtain consistent predictions of ZTA transformation kinetics in vivo? Reported in this paper is a new in toto method for visualizing the surface stability of femoral heads. It is based on CAD-assisted Raman spectroscopy to quantitatively assess the phase transformation observed in ZTA retrievals. Using a series of independent analytical probes, an evaluation of the microscopic mechanisms responsible for the polymorphic transformation is also provided. An outline is given of the possible ways in which the current hydrothermal simulation standard for artificial joints can be improved in an attempt to reduce the gap between in vitro simulation and reality.