Human gastric microbiota transplantation recapitulates premalignant lesions in germ-free mice.

OBJECTIVE: Gastric cancer (GC) is a leading cause of cancer-related mortality. Although microbes besides Helicobacter pylori may also contribute to gastric carcinogenesis, wild-type germ-free (GF) mouse models investigating the role of human gastric microbiota in the process are not yet available. We aimed to evaluate the histopathological features of GF mouse stomachs transplanted with gastric microbiota from patients with different gastric disease states and their relationships with the microbiota. DESIGN: Microbiota profiles in corpus and antrum tissues and gastric fluid from 12 patients with gastric dysplasia or GC were analysed. Thereafter, biopsied corpus and antrum tissues and gastric fluid from patients (n=15 and n=12, respectively) with chronic superficial gastritis, intestinal metaplasia or GC were inoculated into 42 GF C57BL/6 mice. The gastric microbiota was analysed by amplicon sequencing. Histopathological features of mouse stomachs were analysed immunohistochemically at 1 month after inoculation. An independent set of an additional 15 GF mice was also analysed at 1 year. RESULTS: The microbial community structures of patients with dysplasia or GC in the corpus and antrum were similar. The gastric microbiota from patients with intestinal metaplasia or GC selectively colonised the mouse stomachs and induced premalignant lesions: loss of parietal cells and increases in inflammation foci, in F4/80 and Ki-67 expression, and in CD44v9/GSII lectin expression. Marked dysplastic changes were noted at 1 year post inoculation. CONCLUSION: Major histopathological features of premalignant changes are reproducible in GF mice transplanted with gastric microbiota from patients with intestinal metaplasia or GC. Our results suggest that GF mice are useful for analysing the causality of associations reported in human gastric microbiome studies.

WFDC2 Promotes Spasmolytic Polypeptide-Expressing Metaplasia Through the Up-Regulation of IL33 in Response to Injury.

BACKGROUND & AIMS: WAP 4-disulfide core domain protein 2 (WFDC2), also known as human epididymis protein 4, is a small secretory protein that is highly expressed in fibrosis and human cancers, particularly in the ovaries, lungs, and stomach. However, the role of WFDC2 in carcinogenesis is not fully understood. The present study aimed to investigate the role of WFDC2 in gastric carcinogenesis with the use of preneoplastic metaplasia models. METHODS: Three spasmolytic polypeptide-expressing metaplasia (SPEM) models were established in both wild-type and Wfdc2-knockout mice with DMP-777, L635, and high-dose tamoxifen, respectively. To reveal the functional role of WFDC2, we performed transcriptomic analysis with DMP-777-treated gastric corpus specimens. RESULTS: Wfdc2-knockout mice exhibited remarkable resistance against oxyntic atrophy, SPEM emergence, and accumulation of M2-type macrophages in all 3 SPEM models. Transcriptomic analysis revealed that Wfdc2-knockout prevented the up-regulation of interleukin-33 (IL33) expression in the injured mucosal region of SPEM models. Notably, supplementation of recombinant WFDC2 induced IL33 production and M2 macrophage polarization, and ultimately promoted SPEM development. Moreover, long-term treatment with recombinant WFDC2 was able to induce SPEM development. CONCLUSIONS: WFDC2 expressed in response to gastric injury promotes SPEM through the up-regulation of IL33 expression. These findings provide novel insights into the role of WFDC2 in gastric carcinogenesis.

Effects of natural organic matter on the coprecipitation of arsenic with iron.

Natural organic matter (NOM) can affect arsenic speciation and mobility in the environment. In this study, the effects of NOM on the coprecipitation of arsenic with iron were investigated in order to better understand the fate and transport of arsenic in natural environments. The coprecipitation of arsenic with iron was studied in the presence and absence of NOM under various arsenic-to-iron molar ratios (As/Fe) and pH conditions. The addition of humic acid (HA) hindered the As-Fe coprecipitation under high pH and high As/Fe conditions by forming a soluble As-Fe-HA complex. The X-ray diffraction and Fourier transform infrared studies showed that the As-Fe-coprecipitated solid phase was highly affected by pH and As/Fe. The arsenic was coprecipitated with iron as an amorphous ferric arsenate phase at a low pH level or high As/Fe conditions, while the formation of ferrihydrite phase and the arsenic incorporation to the ferrihydrite by adsorption was predominant at high pH levels or low As/Fe conditions. The HA affected the As-Fe-coprecipitated solid phase depending on the As/Fe molar ratio under neutral and alkaline conditions.

Helicobacter pylori promotes epithelial-to-mesenchymal transition by downregulating CK2ß in gastric cancer cells.

Strains of Helicobacter pylori that are positive for the oncoprotein CagA (cytotoxin-associated gene A) are associated with gastric cancer and might be related to the epithelial-to-mesenchymal transition (EMT). Casein kinase 2 (CK2) is a serine/threonine protein kinase that plays a major role in tumorigenesis through signaling pathways related to the EMT. However, the role played by the interaction between CagA and CK2 in gastric carcinogenesis is poorly understood. Although CK2α protein expression remained unchanged during H. pylori infection, we found that CK2α kinase activity was increased in gastric epithelial cells. We also found that the CK2ß protein level decreased in H. pylori-infected gastric cancer cells in CagA-dependent manner and demonstrated that CagA induced CK2ß degradation via HDM2 (human double minute 2; its murine equivalent is MDM2). We observed that CagA induced HDM2 protein phosphorylation and that p53 levels were decreased in H. pylori-infected gastric cancer cells. In addition, downregulation of CK2ß induced AKT Ser473 phosphorylation and decreased the AKT Ser129 phosphorylation level in gastric cancer cells. We also found that the downregulation of CK2ß triggered the upregulation of Snail levels in gastric cancer cells. Furthermore, our in vivo experiments and functional assays of migration and colony formation suggest that CK2ß downregulation is a major factor responsible for the EMT in gastric cancer. Therefore, CK2 could be a key mediator of the EMT in H. pylori-infected gastric cancer and could serve as a molecular target for gastric cancer treatment.

Evaluation of infertility efficacy of the E. coli expressed STF2-GnRH vaccine in male cats.

Gonadotropin-releasing hormone (GnRH) is secreted from the hypothalamus and anti-GnRH antibodies are not formed under normal conditions. However, administration an excess of recombinant GnRH protein results in the formation of anti-GnRH. We evaluated the efficacy of the recombinant Salmonella typhimurium flagellin fljB (STF2)-GnRH vaccine in inducing infertility in 17 intact male cats. The first vaccination and a boosting vaccine was injected for examination. Serum was obtained from blood collected at monthly intervals and anti-GnRH antibodies and testosterone concentrations were determined. Six months after the vaccination, testicular samples are obtained and used for histological examination. Compared with sham control group, the injection groups showed an increase in anti-GnRH antibody titers and testosterone concentrations tended to be reduced in the injection groups and increased in the control group. Histological evaluations and Johnsen's testicular biopsy scores revealed testicular hypoplasia in the 2 injection groups. Consequently, normal sexual maturation with sperm production was observed in the control group. In contrast, the cats that received the GnRH vaccine showed weak (2 of 7 cats) or moderate (4 out of 7 cats) dose-dependent infertility effects. On the basis of the results, the STF2-GnRH vaccine was identified to be effective in inducing infertility in male cats. The results of this study thus indicate the possibility of immunological castration targeting feral cats.

Protective effect of pterostilbene on testicular ischemia/reperfusion injury in rats.

PURPOSE: To evaluate protective effect of pterostilbene against testicular ischemia/reperfusion (I/R) injury, which results in increased formation of oxidative stress, leading to testicular apoptosis and impaired spermatogenesis. METHODS: Thirty two pubertal male Sprague-Dawley rats weighing 180-220g were selected and randomly divided into the following four groups: group A (normal control group), group B (sham-operated group), group C (induced I/R injury group), group D (induced I/R injury group receiving pterostilbene treatment). Johnsen's scores and mean seminiferous tubule diameters were evaluated for histopathologic assessment; germinal cell apoptosis was evaluated by the transferase dUTP nick end labeling (TUNEL) assay and immunohistochemistry for caspases. Malondialdehyde (MDA) levels were assessed as an indicator of oxidative stress and total antioxidant capacity (TAC) was measured. RESULTS: Germ cell apoptosis and MDA level significantly increased whereas TAC significantly decreased in group C; moreover, abnormal morphology and impaired spermatogenesis were observed in group C. In contrast, treatment with pterostilbene inhibited lipid peroxidation and apoptosis induced by ROS and restored the antioxidant capacity in group D. CONCLUSIONS: These results show that treatment with pterostilbene may be a promising therapy for testicular I/R injury.

Extractive and oxidative removal of copper bound to humic acid in soil.

Copper (Cu) is often found strongly bound to natural organic matter (NOM) in soil through the formation of strong Cu-NOM complexes. Therefore, in order to successfully remediate Cu-contaminated soils, effective removal of Cu bound to soil organic matter should be considered. In this study, we investigated soil washing methods for Cu removal from a synthetic Cu-contaminated model silica soil coated with humic acid (HA) and from field contaminated soil. Various reagents were studied to extract Cu bound to NOM, which included oxidant (H2O2), base (NaOH), and chelating agents (citric acid and ethylenediaminetetraacetic acid (EDTA)). Among the wash reagents, EDTA extracted Cu most effectively since EDTA formed very strong complexes with Cu, and Cu-HA complexes were transformed into Cu-EDTA complexes. NaOH extracted slightly less Cu compared to EDTA. HA was effectively extracted from the model soil under strongly alkaline conditions with NaOH, which seemed to concurrently release Cu bound to HA. However, chemical oxidation with H2O2 was not effective at destroying Cu-HA complexes. Fourier transform infrared spectroscopy and elemental analysis revealed that chelating agents such as citrate and EDTA were adsorbed onto the model soil via possible complexation between HA and extraction agents. The extraction of Cu from a field contaminated soil sample was effective with chelating agents, while oxidative removal with H2O2 and extractive removal with NaOH separated negligible amounts of Cu from the soil. Based on these results, Cu bound to organic matter in soil could be effectively removed by chelating agents, although remnant agents may remain in the soil.

Environmental assessment on electrokinetic remediation of multimetal-contaminated site: a case study.

In this study, an environmental assessment on an electrokinetic (EK) system for the remediation of a multimetal-contaminated real site was conducted using a green and sustainable remediation (GSR) tool. The entire EK process was classified into major four phases consisting of remedial investigations (RIs), remedial action construction (RAC), remedial action operation (RAO), and long-term monitoring (LTM) for environmental assessment. The environmental footprints, including greenhouse gas (GHG) emissions, total energy used, air emissions of criteria pollutants, such as NOx, SOx, and PM10, and water consumption, were calculated, and the relative contribution in each phase was analyzed in the environmental assessment. In the RAC phase, the relative contribution of the GHG emissions, total energy used, and PM10 emissions were 77.3, 67.6, and 70.4%, respectively, which were higher than those of the other phases because the material consumption and equipment used for system construction were high. In the RAO phase, the relative contributions of water consumption and NOx and SOx emissions were 94.7, 85.2, and 91.0%, respectively, which were higher than those of the other phases, because the water and electricity consumption required for system operation was high. In the RIs and LTM phases, the environmental footprints were negligible because the material and energy consumption was less. In conclusion, the consumable materials and electrical energy consumption might be very important for GSR in the EK remediation process, because the production of consumable materials and electrical energy consumption highly affects the GHG emissions, total energy used, and air emissions such as NOx and SOx.

Environmental assessment on a soil washing process of a Pb-contaminated shooting range site: a case study.

In this study, an environmental assessment on a soil washing process for the remediation of a Pb-contaminated shooting range site was conducted, using a green and sustainable remediation tool, i.e., SiteWise ver. 2, based on data relating specifically to the actual remediation project. The entire soil washing process was classified into four major stages, consisting of soil excavation (stage I), physical separation (stage II), acid-based (0.2 N HCl) chemical extraction (stage III), and wastewater treatment (stage IV). Environmental footprints, including greenhouse gas (GHG) emissions, energy consumption, water consumption, and critical air pollutant productions such as PM10, NO x , and SO x , were calculated, and the relative contribution of each stage was analyzed in the environmental assessment. In stage I, the relative contribution of the PM10 emissions was 55.3 % because the soil excavation emitted the fine particles. In stage II, the relative contribution of NO x and SO x emissions was 42.5 and 52.5 %, respectively, which resulted from electricity consumption for the operation of the separator. Stage III was the main contributing factor to 63.1 % of the GHG emissions, 67.5 % of total energy used, and 37.4 % of water consumptions. The relatively high contribution of stage III comes from use of consumable chemicals such as HCl and water-based extraction processes. In stage IV, the relative contributions of GHG emissions, total energy used, and NO x and SO x emissions were 23.2, 19.4, 19.5, and 25.3 %, respectively, which were caused by chemical and electricity demands for system operation. In conclusion, consumable chemicals such as HCl and NaOH, electric energy consumption for system operation, and equipment use for soil excavation were determined to be the major sources of environmental pollution to occur during the soil washing process. Especially, the acid-based chemical extraction process should be avoided in order to improve the sustainability of soil washing processes.

Sexual activity counteracts the suppressive effects of chronic stress on adult hippocampal neurogenesis and recognition memory.

Adult neurogenesis can be influenced by a variety of factors. Stress is one of the most potent inhibitors of hippocampal neurogenesis. Stress effects on adult hippocampal neurogenesis are affected differently by environmental factors, including social interaction. Sexual behavior between males and females in a social context has been suggested to influence neurogenesis and enhance hippocampal cell proliferation. However, the mechanisms of action of sexual interaction, the possible changes relative to stress state, and its effects on learning and memory remain uncertain. The current study examined the influence of sexual interaction on neurological responses in adult male mice and the function of sexual interaction relative to recognition memory in stress states. Changes in the expression of neurotrophic and transcription factors were assessed in reference to stress and/or sexual behaviors. The survival of newly generated cells and their rate of differentiation into neurons were determined in the hippocampus of chronically stressed and/or sexually experienced mice. Finally, to evaluate whether sexual experience alters adult hippocampal function, we tested learning and memory in a recognition memory task. The results demonstrated that sexual activity increased the expression of brain-derived neurotrophic factor, tyrosine kinase B, and cAMP response element-binding factor. Furthermore, the results supported the view that sexual interaction could be helpful for buffering adult hippocampal neurogenesis and recognition memory function against the suppressive actions of chronic stress.