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1.  Plant diversity effects on grassland productivity are robust to both nutrient enrichment and drought 
Global change drivers are rapidly altering resource availability and biodiversity. While there is consensus that greater biodiversity increases the functioning of ecosystems, the extent to which biodiversity buffers ecosystem productivity in response to changes in resource availability remains unclear. We use data from 16 grassland experiments across North America and Europe that manipulated plant species richness and one of two essential resources—soil nutrients or water—to assess the direction and strength of the interaction between plant diversity and resource alteration on above-ground productivity and net biodiversity, complementarity, and selection effects. Despite strong increases in productivity with nutrient addition and decreases in productivity with drought, we found that resource alterations did not alter biodiversity–ecosystem functioning relationships. Our results suggest that these relationships are largely determined by increases in complementarity effects along plant species richness gradients. Although nutrient addition reduced complementarity effects at high diversity, this appears to be due to high biomass in monocultures under nutrient enrichment. Our results indicate that diversity and the complementarity of species are important regulators of grassland ecosystem productivity, regardless of changes in other drivers of ecosystem function.
PMCID: PMC4843698  PMID: 27114579
plant diversity; global change drivers; resource amendment; resource reduction; soil nutrients; drought
2.  Active foraging for toxic prey during gestation in a snake with maternal provisioning of sequestered chemical defences 
Many animals sequester dietary defensive compounds and incorporate them into the offspring, which protects the young against predation. One possible but poorly investigated question is whether females of such species actively prey upon toxic diets. The snake Rhabdophis tigrinus sequesters defensive steroids from toads consumed as prey; it also feeds on other amphibians. Females produce chemically armed offspring in direct proportion to their own level of toad-derived toxins by provisioning the toxins to their eggs. Our field observations of movements and stomach contents of radio-tracked R. tigrinus showed that gravid snakes preyed upon toads by actively foraging in the habitat of toads, even though toads were a scarce resource and toad-searching may incur potential costs. Our Y-maze experiments demonstrated that gravid females were more likely to trail the chemical cues of toads than were males or non-gravid females. These results showed behavioural switching in females and active foraging for scarce, toxic prey during gestation. Because exploitation of toads by gravid females results in their offspring being more richly endowed with prey-derived toxins, active foraging for toxic prey is expected to be an adaptive antipredator trait, which may enhance chemical defence in offspring.
PMCID: PMC4262175  PMID: 25392472
antipredator defence; maternal care; defensive sequestration; prey choice; foraging strategy; Rhabdophis tigrinus
3.  Academic inequality through the lens of community ecology: a meta-analysis 
PeerJ  2015;3:e1457.
Ecological assemblages are generally characterized by a few dominant species and numerous others. Such unequal distributions of dominance also emerge in human society, including in scientific communities. Here, based on formal community ecological analyses, we show the temporal trends in the number of scientific publication in the discipline of “ecology.” Based on this, we infer possible factors causing the imbalance of reputation and dominance among countries. We relied on 454 ecological meta-analysis papers published from 1998 to 2014, which sourced over 29,000 original publications. Formal meta-analyses are essential for synthesizing findings from individual studies and are critical for assessing issues and informing policy. We found that, despite the rapid expansion of outlets for ecology papers (analogous to an increase in carrying capacity, in ecological systems), country diversity as determined from first author affiliations (analogous to species diversity) did not increase. Furthermore, a country identity was more powerful than the popularity of the scientific topic and affected the chance of publication in high-profile journals, independent of the potential novelty of findings and arguments of the papers, suggesting possible academic injustice. Consequently, a rank order and hierarchy has been gradually formed among countries. Notably, this country-dominance rank is not only specific to this scientific domain but also universal across different societal situations including sports and economics, further emphasizing that inequality and hierarchical structure exist even in modern human society. Our study demonstrates a need for having robust frameworks to facilitate equality and diversity in the scientific domain in order to better inform society and policy.
PMCID: PMC4671160  PMID: 26644987
Publication bias; Dominance; Societal hierarchy; Meta-analysis; Species abundance distributions
4.  Potential envenomation by the aglyphous pseudoxyrhophiine snake Leioheterodon madagascariensis and description of its dentition 
We report on a case of potential envenomation caused by multiple bites by the aglyphous opisthodont snake Leioheterodon madagascariensis in the left thumb of a healthy adult man, which is among the most serious snakebites hitherto reported from Madagascar. The adult snake (total length > 1 meter) was unusually aggressive before and during capture. The symptoms included extensive bleeding, severe local pain, and substantial swelling of the hand and the distal part of the lower arm. The swelling disappeared entirely after five days, but pain in the thumb (when moved) was recognizable even longer. Although L. madagascariensis is widespread and common in anthropogenic habitats in eastern and western Madagascar, this case report seems to be the first description of long-lasting symptoms of its bite. Since aglyphous snakes are relatively rarely involved in “envenomation” and because hemolytic activity has been recorded in the secretions of the Duvernoy’s glands of Leioheterodon, we describe its dentition using microcomputed tomography and discuss the potential mode of envenomation in this case.
Electronic supplementary material
The online version of this article (doi:10.1186/s40409-015-0047-2) contains supplementary material, which is available to authorized users.
PMCID: PMC4654879  PMID: 26594226
Madagascar; Lamprophiidae; Pseudoxyrhophiinae; Leioheterodon madagascariensis; Envenomation; Dentition; Microcomputed tomography
5.  Changes in Surgical Site Infections after Living Donor Liver Transplantation 
PLoS ONE  2015;10(8):e0136559.
Surgical site infections (SSIs) are a major threat for liver transplant recipients. We prospectively studied SSIs after living donor liver transplantation (LDLT) at Kyoto University Hospital from April 2001 to March 2002 (1st period) and from January 2011 to June 2012 (2nd period). We investigated the epidemiology of SSIs after LDLT and determined the differences between the two periods. A total of 129 adult recipients (66 during the 1st period and 63 during the 2nd period) and 72 pediatric recipients (39 and 33) were included in this study. The SSI rates for each period were 30.3% (1st period) and 41.3% (2nd period) among the adult recipients and 25.6% and 30.3% among the pediatric recipients. The overall rates of 30-day mortality among adult transplant recipients with SSIs were 10.0% (1st period) and 3.9% (2nd period). No pediatric recipient died from SSIs after LDLT in either period. The incidence of Enterococcus faecium increased from 5.0% to 26.9% in the adults and from 10.0% to 40.0% in the pediatric patients. Extended-spectrum β-lactamase-producing Enterobacteriaceae were emerging important isolates during the 2nd period. For this period, a univariate analysis showed that ABO incompatibility (P = 0.02), total operation duration (P = 0.01), graft-to-recipient body weight ratio (GRWR [P = 0.04]), and Roux-en-Y biliary reconstruction (P<0.01) in the adults and age (P = 0.01) and NHSN risk index (P = 0.02) in the children were associated with SSI development. In a multivariate analysis, lower GRWR (P = 0.02) and Roux-en-Y biliary reconstruction (P<0.01) in the adults and older age (P = 0.01) in the children were independent risk factors for SSIs during the 2nd period. In conclusion, SSIs caused by antibiotic resistant bacteria may become a major concern. Lower GRWR and Roux-en-Y biliary reconstruction among adult LDLT recipients and older age among pediatric LDLT recipients increased the risk of developing SSIs after LDLT.
PMCID: PMC4556480  PMID: 26322891
6.  Long-Term Outcomes of Hepatic Resection versus Living Donor Liver Transplantation for Hepatocellular Carcinoma: A Propensity Score-Matching Study 
Disease Markers  2015;2015:425926.
Hepatic resection (HR) and liver transplantation (LT) are surgical treatment options for hepatocellular carcinoma (HCC). However, it is clinically impossible to perform a randomized, controlled study to determine the usefulness of these treatments. The present study compared survival rates and recurrence rates of HR versus living donor LT (LDLT) for HCC by using the propensity score method. Between January 1999 and August 2012, 936 patients (732 HR, 204 LDLT) underwent surgical therapy for HCC in our center. Using the propensity score matching, 80 well-balanced patients were defined. The 1- and 5-year overall survival rates were 90% and 53% in the HR group and 82% and 63% in the LT group, respectively. They were not significantly different between the two groups. The odds ratio estimated using the propensity score matching analysis was 0.842 (P = 0.613). The 1- and 5-year recurrence rates were significantly lower in the LT group (9% and 21%) than in the HR group (43% and 74%) (P < 0.001), and the odds ratio was 0.214 (P = 0.001). In conclusion, HR should be considered a valid alternative to LDLT taking into consideration the risk for the living donor based on the results of this propensity score-matching study.
PMCID: PMC4397476  PMID: 25922554
7.  Topographic patterns in the phylogenetic structure of temperate forests on steep mountainous terrain 
AoB Plants  2015;7:plv134.
In this study, we tested the topographic trends of measurements of phylogenetic community structure in a catchment covered by temperate forests with complex relief in Japan. We found that phylogenetic structure changed with increasing slope inclination, change of slope aspect from south to north, and decreasing soil depth. Our results suggested that environmental filtering caused by steep topography tended to restrict phylogenetic community composition at relatively stressful sites. On the other hand, species interaction functioned more strongly at relatively stress-free sites.
In rugged terrain subject to active geomorphological processes, the species composition of forest communities changes along topographic gradients over short distances. However, the phylogenetic structure of forests on rugged terrain has rarely been examined. Understanding such structures provides insight into community assembly rules dependent on local environmental conditions. To this end, we tested the topographic trends of measurements of phylogenetic community structure [net relatedness index (NRI) and nearest taxon index] in a catchment covered by temperate forests with complex relief in Japan. We found that phylogenetic structure changed from over-dispersion to clustering with increasing slope inclination, change of slope aspect from south to north and decreasing soil depth. This result suggested that environmental filtering tended to restrict community composition at relatively stressful sites, whereas species interaction functioned more strongly at relatively stress-free sites. Relatively stressful sites were characterized by early-successional species that tended to assemble in certain phylogenetic clades, whereas highly competitive late-successional species associated with lower NRI at relatively stress-free sites. However, despite this significant topographic tendency, phylogenetic community structures were not statistically different from random assumptions in most plots. This obscuration of the phylogenetic structures at the plot level could be interpreted as species adapting to disturbances because they were abundant in this catchment. Accordingly, we suggest that a stochastic process was also important for this community at the plot level, although biotic and abiotic environmental filtering controlled the vegetation structure in the catchment, where disturbances occur frequently because of geomorphological mountain denudation processes owing to active crustal movements and abundant rainfall.
PMCID: PMC4683994  PMID: 26602986
Community assembly rule; heterogeneous environmental condition; phylogenetic diversity; temperate forest in East Asia; topographic gradients
8.  Sequestered defensive toxins in tetrapod vertebrates: principles, patterns, and prospects for future studies 
Chemoecology  2012;22(3):141-158.
Chemical defenses are widespread among animals, and the compounds involved may be either synthesized from nontoxic precursors or sequestered from an environmental source. Defensive sequestration has been studied extensively among invertebrates, but relatively few examples have been documented among vertebrates. Nonetheless, the number of described cases of defensive sequestration in tetrapod vertebrates has increased recently and includes diverse lineages of amphibians and reptiles (including birds). The best-known examples involve poison frogs, but other examples include natricine snakes that sequester toxins from amphibians and two genera of insectivorous birds. Commonalities among these diverse taxa include the combination of consuming toxic prey and exhibiting some form of passive defense, such as aposematism, mimicry, or presumptive death-feigning. Some species exhibit passive sequestration, in which dietary toxins simply require an extended period of time to clear from the tissues, whereas other taxa exhibit morphological or physiological specializations that enhance the uptake, storage, and/or delivery of exogenous toxins. It remains uncertain whether any sequestered toxins of tetrapods bioaccumulate across multiple trophic levels, but multitrophic accumulation seems especially likely in cases involving consumption of phytophagous or mycophagous invertebrates and perhaps consumption of poison frogs by snakes. We predict that additional examples of defensive toxin sequestration in amphibians and reptiles will be revealed by collaborations between field biologists and natural product chemists. Candidates for future investigation include specialized predators on mites, social insects, slugs, and toxic amphibians. Comprehensive studies of the ecological, evolutionary, behavioral, and regulatory aspects of sequestration will require teams of ecologists, systematists, ethologists, physiologists, molecular biologists, and chemists. The widespread occurrence of sequestered defenses has important implications for the ecology, evolution, and conservation of amphibians and reptiles.
PMCID: PMC3418492  PMID: 22904605
Sequestration; Dietary toxins; Antipredator defense; Aposematism; Amphibians; Reptiles
9.  Vigilance against predators induced by eavesdropping on heterospecific alarm calls in a non-vocal lizard Oplurus cuvieri cuvieri (Reptilia: Iguania) 
Prey animals can reduce their risk of predation by detecting potential predators before encounters occur. Some animals gain information about nearby predators by eavesdropping on heterospecific alarm calls. Despite having well-developed ears, most lizards do not use vocal information for intraspecific communication, and few studies have shown practical use of the ears in wild lizards. Here, we show that the Madagascan spiny-tailed iguana (Oplurus cuvieri cuvieri) obtains auditory signals for predator detection. The Madagascan spiny-tailed iguana and the Madagascar paradise flycatcher (Terpsiphone mutata) are syntopic inhabitants of the Ampijoroa dry deciduous forest of Madagascar. The iguana and the flycatcher have neither a predator–prey relationship nor resource competition, but they have shared predators such as raptors and snakes. Using playback experiments, we demonstrated that the iguana discriminates mobbing alarm calls of the flycatcher from its songs and then enhances its vigilance behaviour. Our results demonstrate the occurrence of an asymmetrical ecological relationship between the Madagascan spiny-tailed iguana and the paradise flycatcher through eavesdropping on information about the presence of predators. This implies that indirect interspecific interactions through information recognition may be more common than generally thought in an animal community.
PMCID: PMC2842824  PMID: 20031993
eavesdropping; heterospecific signal; mobbing alarm call; asymmetrical relationship; Madagascan spiny-tailed iguana
10.  Thrombotic microangiopathy-like disorder after living-donor liver transplantation: A single-center experience in Japan 
AIM: To investigate thrombotic microangiopathy (TMA) in liver transplantion, because TMA is an infrequent but life-threatening complication in the transplantation field.
METHODS: A total of 206 patients who underwent living-donor liver transplantation (LDLT) were evaluated, and the TMA-like disorder (TMALD) occurred in seven recipients.
RESULTS: These TMALD recipients showed poor outcomes in comparison with other 199 recipients. Although two TMALD recipients successfully recovered, the other five recipients finally died despite intensive treatments including repeated plasma exchange (PE) and re-transplantation. Histopathological analysis of liver biopsies after LDLT revealed obvious differences according to the outcomes. Qualitative analysis of antibodies against a disintegrin-like domain and metalloproteinase with thrombospondin type 1 motifs (ADAMTS-13) were negative in all patients. The fragmentation of red cells, the microhemorrhagic macules and the platelet counts were early markers for the suspicion of TMALD after LDLT. Although the absolute values of von Willebrand factor (vWF) and ADAMTS-13 did not necessarily reflect TMALD, the vWF/ADAMTS-13 ratio had a clear diagnostic value in all cases. The establishment of adequate treatments for TMALD, such as PE for ADAMTS-13 replenishment or treatments against inhibitory antibodies, must be decided according to each case.
CONCLUSION: The optimal induction of adequate therapies based on early recognition of TMALD by the reliable markers may confer a large advantage for TMALD after LDLT.
PMCID: PMC3080720  PMID: 21528059
Thrombotic microangiopathy; Liver transplantation; von Willebrand factor; A disintegrin-like domain and metalloproteinase with thrombospondin type 1 motifs; Complication
11.  Activating transcription factor-3 (ATF3) functions as a tumor suppressor in colon cancer and is up-regulated upon heat-shock protein 90 (Hsp90) inhibition 
BMC Cancer  2010;10:668.
Activating transcription factor-3 (ATF3) is involved in the complex process of cellular stress response. However, its exact role in cancer is discussed controversially because both tumor suppressive and oncogenic effects have been described. Here we followed-up on our previous observation that inhibition of Hsp90 may increase ATF3 expression and sought to determine the role of ATF3 in colon cancer.
Regulation of ATF3 was determined in cancer cells using signaling inhibitors and a heat-shock protein-90 (Hsp90) antagonist. Human HCT116 cancer cells were stably transfected with an ATF3-shRNA or a luciferase-shRNA expression plasmid and alterations in cell motility were assessed in migration assays. The impact of ATF3 down-regulation on cancer growth and metastasis were investigated in a subcutaneous tumor model, a model of hepatic tumor growth and in a model of peritoneal carcinomatosis. Human colon cancer tissues were analyzed for ATF3 expression.
The results show that therapeutic Hsp90 inhibition substantially up-regulates the expression of ATF3 in various cancer cells, including colon, gastric and pancreatic cancer. This effect was evident both in vitro and in vivo. RNAi mediated knock-down of ATF3 in HCT116 colon cancer cells significantly increased cancer cell migration in vitro. Moreover, in xenogenic mouse models, ATF3 knock-down promoted subcutaneous tumor growth and hepatic metastasis, as well as peritoneal carcinomatosis. Importantly, ATF3 expression was lower in human colon cancer specimens, as compared to corresponding normal surrounding tissues, suggesting that ATF3 may represent a down-regulated tumor suppressor in colon cancer.
In conclusion, ATF3 down-regulation in colon cancer promotes tumor growth and metastasis. Considering that blocking Hsp90 induces ATF3 expression, Hsp90 inhibition may represent a valid strategy to treat metastatic colon cancer by up-regulating this anti-metastatic transcription factor.
PMCID: PMC3003660  PMID: 21129190
12.  ENMD-1198, a novel tubulin-binding agent reduces HIF-1alpha and STAT3 activity in human hepatocellular carcinoma(HCC) cells, and inhibits growth and vascularization in vivo 
BMC Cancer  2008;8:206.
Hepatocellular carcinoma (HCC) represents a highly vascularized tumor entity and the process of angiogenesis is essential for the growth of HCC. Importantly, the pro-angiogenic transcription factors HIF-1α and STAT3 have been implicated in HCC progression, thus representing interesting targets for molecular targeted therapy. We hypothesized that therapeutic inhibition of HIF-1α could be achieved by using a novel tubulin-binding agent (ENMD-1198). ENMD-1198 is an analog of 2-methoxyestradiol (2ME2) with antiproliferative and antiangiogenic activity.
The human HCC cell lines HUH-7 and HepG2 were used for experiments. Effects of ENMD-1198 on constitutive and inducible (hypoxia, growth factors) activation of signaling cascades, including HIF-1α and STAT3, were investigated by Western blotting. Changes in VEGF expression were determined by real-time PCR. Effects of ENMD-1198 on cancer cell migration and invasion were evaluated in in vitro-assays. The growth-inhibitory effects of ENMD-1198 (200 mg/kg/day) were determined in a subcutaneous tumor model (HUH-7).
ENMD-1198 inhibited the phosphorylation of MAPK/Erk, PI-3K/Akt and FAK. Moreover, activation of HIF-1α and STAT3 was dramatically reduced by ENMD-1198, which resulted in lower VEGF mRNA expression (P < 0.05). In addition, tumor cell migratory and invasive properties were significantly inhibited (P < 0.05, for both). In vivo, treatment with ENMD-1198 led to a significant reduction in tumor growth, tumor vascularization, and numbers of proliferating tumor cells (P < 0.05 for all).
The novel microtubule destabilizing agent ENMD-1198 is suitable for inhibiting HIF-1α and STAT3 in human HCC cells and leads to reduced tumor growth and vascularization in vivo. Hence, inhibition of HIF-1α and STAT3 could prove valuable for therapy of hepatocellular carcinoma.
PMCID: PMC2496914  PMID: 18651980
13.  Shoot Development and Extension of Quercus serrata Saplings in Response to Insect Damage and Nutrient Conditions 
Annals of Botany  2006;98(1):219-226.
• Background and Aims Plants have the ability to compensate for damage caused by herbivores. This is important to plant growth, because a plant cannot always avoid damage, even if it has developed defence mechanisms against herbivores. In previous work, we elucidated the herbivory-induced compensatory response of Quercus (at both the individual shoot and whole sapling levels) in both low- and high-nutrient conditions throughout one growing season. In this study, we determine how the compensatory growth of Quercus serrata saplings is achieved at different nutrient levels.
• Methods Quercus serrata saplings were grown under controlled conditions. Length, number of leaves and percentage of leaf area lost on all extension units (EUs) were measured.
• Key Results Both the probability of flushing and the length of subsequent EUs significantly increased with an increase in the length of the parent EU. The probability of flushing increased with an increase in leaf damage of the parent EU, but the length of subsequent EUs decreased. This indicates that EU growth is fundamentally regulated at the individual EU level. The probabilities of a second and third flush were significantly higher in plants in high-nutrient soil than those in low-nutrient soil. The subsequent EUs of damaged saplings were also significantly longer at high-nutrient conditions.
• Conclusions An increase in the probability of flushes in response to herbivore damage is important for damaged saplings to produce new EUs; further, shortening the length of EUs helps to effectively reproduce foliage lost by herbivory. The probability of flushing also varied according to soil nutrient levels, suggesting that the compensatory growth of individual EUs in response to local damage levels is affected by the nutrients available to the whole sapling.
PMCID: PMC2803554  PMID: 16709576
Quercus serrata; extension units; compensation; soil fertility; herbivore; sapling; rhythmic growth; flush; leaf damage
14.  New Model for Studying the Migration of Immune Cells into Intestinal Epithelial Cell Monolayers 
Cytotechnology  2003;43(1-3):57-64.
A novel cell culture system was constructed to analyze the direct interaction between intestinal epithelial cells and immune cells. Human intestinal epithelial Caco-2 cells were monolayer-cultured on the under side of a permeable membrane (12 μm pore size) in a Millicell insert. Integrated monolayers of Caco-2 cells had formed after 12 days of culture. Human monocyte/macrophage-like THP-1 cells were then added to the upper chamber of the insert, and their migration into the Caco-2 cell monolayers was observed by confocal laser scanning microscopy, after staining the cells with specific antibodies. When MCP-1, a β-chemokine, was added to the apical side of the monolayer, a greater number of THP-1 cells migrated into the Caco-2 cell monolayers. This cell culture system will be useful for studying the behavior of macrophages in the intestinal epithelial cell monolayers at the initial stage of an intestinal immune reaction.
PMCID: PMC3449588  PMID: 19003208
Caco-2; cell migration; chemokine; intestinal epithelium; THP-1

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