Transcriptomic Responses Of Corpuscle Of Stannius Gland Of Japanese Eels (anguilla Japonica) To Changes In Water Salinity

Physiological studies of a unique endocrine gland in fish, named corpuscles of Stannius (CS), described a Ca2+-regulatory function for this gland mediated by stanniocalcin-1, a hypocalcemic polypeptide hormone. However, to date, the endocrine functions of the glands have not been completely elucidated. We hypothesized that other unidentified active principles in the glands are involved in the regulation of plasma ion (Na+, Ca2+) and/or blood pressure. In this study, transcriptome sequencing of CS glands was performed using Japanese eels (Anguilla japonica) adapted to freshwater (FW) or seawater (SW) to reveal the presence and differential expression of genes encoding proteins related to the ion-osmoregulatory and pressor functions. We acquired a total of 14.1 Mb and 12.1 Mb quality-trimmed reads from the CS glands collected from FW and SW adapted eels, respectively. The de novo assembly resulted in 9254 annotated genes. Among them, 475 genes were differentially expressed with 357 up- and 118 down-regulated in the SW group. Gene ontology analysis further demonstrated the presence of natriuresis and pressor related genes. In summary, ours is the first study using high-throughput sequencing to identify gene targets that could explain the physiological importance of the CS glands.published_or_final_versio

A prospective interventional study to examine the effect of a silver alloy and hydrogel--coated catheter on the incidence of catheter-associated urinary tract infection

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Tissue-specific transcriptome assemblies of the marine medaka Oryzias melastigma and comparative analysis with the freshwater medaka Oryzias latipes

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Use of interferon gamma release assay to assess latent tuberculosis infection among healthcare workers in Hong Kong

Key Messages 1. Overall baseline interferon gamma release assay positivity was 20.7%. 2. The conversion to interferon gamma release assay positivity at 3 months was 8.85% in the exposed group and 4.54% in the non-exposed group using the conventional cut-off of 0.35 IU/mL. 3. When grey zone results (0.2I-0.7 IU/mL) were included, the proportion of non-specific conversions and reversions could be reduced. 4. Interferon gamma release assay can be an adjunct tool in contact investigation of latent tuberculosis infection in healthcare workers.published_or_final_versio

Identification and characterization of tropomyosin 3 associated with granulin-epithelin precursor in human hepatocellular carcinoma

Background and Aim: Granulin-epithelin precursor (GEP) has previously been reported to control cancer growth, invasion, chemo-resistance, and served as novel therapeutic target for cancer treatment. However, the nature and characteristics of GEP interacting partner remain unclear. The present study aims to identify and characterize the novel predominant interacting partner of GEP using co-immunoprecipitation and mass spectrometry. Methods and Results: Specific anti-GEP monoclonal antibody was used to capture GEP and its interacting partner from the protein extract of the liver cancer cells Hep3B. The precipitated proteins were analyzed by SDS-PAGE, followed by mass spectrometry and the protein identity was demonstrated to be tropomyosin 3 (TPM3). The interaction has been validated in additional cell models using anti-TPM3 antibody and immunoblot to confirm GEP as the interacting partner. GEP and TPM3 expressions were then examined by real-time quantitative RT-PCR in clinical samples, and their transcript levels were significantly correlated. Elevated TPM3 levels were observed in liver cancer compared with the adjacent non-tumorous liver, and patients with elevated TPM3 levels were shown to have poor recurrence-free survival. Protein expression of GEP and TPM3 was observed only in the cytoplasm of liver cancer cells by immunohistochemical staining. Conclusions: TPM3 is an interacting partner of GEP and may play an important role in hepatocarcinogenesis. © 2012 Lam et al.published_or_final_versio

Modulation of AhR-mediated CYP1A1 mRNA and EROD activities by 17β-estradiol and dexamethasone in TCDD-induced H411E cells

TCDD elicits a variety of species- and organ-specific pathological consequences. The differential toxicities are thought to relate to the de novo modulation of TCDD action by endogenous hormones. Previous studies from this laboratory demonstrated a dose-and time-dependent induction of CYP1A1 expression and 7-ethoxyresorufin-O-deethylase (EROD) activities in H4IIE cells by picomolar levels of TCDD treatment. In this study, we examined the hormonal modulation of TCDD-elicited AhR-mediated biochemical responses. Lipid-soluble hormones, 17β-estradiol (E2), diethylstilbestrol (DES), testosterone (T), 5α-dihydrotestosterone (DHT), dexamethasone (DEX), and T3 were studied for their possible interactions with the TCDD-mediated effects. Our results showed that CYP1A1 expression and EROD activities induced by TCDD were potentiated or suppressed, respectively, by DEX or E2/DES treatment. Other tested hormones, however, had no significant effect. Using a receptor antagonist (RU486), DEX-mediated potentiation of TCDD-elicited EROD activity was completely abolished. E2-mediated suppression, however, was not affected by cotreatment with the estrogen receptor antagonists, 4-hydroxytamoxifen or ICI 182780. Taking a step further to dissect the possible mechanisms involved, with the aid of cycloheximide (CHX), DEX-mediated potentiation was found to depend on the posttranscriptional process. The DEX pretreatment study indicated that the potentiation was a time-dependent process. In contrast, E2-mediated suppression did not rely on the synthesis of protein factors. Presumably it might hinder the formation of the activated TCDD/AhR complex and so the subsequent binding on DRE. © Society of Toxicology 2004; all rights reserved.link_to_subscribed_fulltex

Effects of TCDD in modulating the expression of Sertoli cell secretory products and markers for cell-cell interaction

Among different mammalian tissues, testis is found to be one of the most sensitive organs to TCDD exposure. In this study, primary Sertoli cell culture was established. The purity of the cultured cells was verified using 3β-hydroxysteroid dehydrogenase, alkaline phosphatase as well as testosterone induction assays. Effects of TCDD in modulating the expression of CYP1A1, aromatase, secretory products (i.e. Müllerian inhibiting substance (MIS), 17β-estradiol (E 2) and lactate) and markers for cell-cell interaction (i.e. sertolin and testin) were then examined. Our data demonstrated that Sertoli cells exposed to 0.2-2000 pg/ml of TCDD showed a dose dependent induction of CYP1A1 mRNA. The minimal dose of activation was 2 pg/ml, which indicated that the cell was very sensitive to TCDD exposure. However, there was little or no detectable level CYP1A1 protein and EROD activities found. Dose-dependent inductions of aromatase transcript (200%) and E 2 (20%) secretion were measured. In addition there was a significant reduction (40%) of MIS mRNA. No detectable change in the level of secreted lactate was observed. Sertolin and testin, the gene makers for cell-cell interactions were differentially modulated upon TCDD treatment. Taken together, the results implicated that TCDD exposure might interfere with the normal Sertoli cell functions. © 2004 Elsevier Ireland Ltd. All rights reserved.link_to_subscribed_fulltex

Inhibition of CYP450scc expression in dioxin-exposed rat Leydig cells

Polychlorinated dibenzo-p-dioxins, such as 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) have been recognized as highly potent developmental and reproductive toxins. We have previously demonstrated effects of TCDD in modulating the expression of rat Sertoli cell secretory products and markers for cell-cell interaction. In this study, we examined the direct biological effects of TCDD in rat Leydig cell primary cultures. Mature rat Leydig cells were purified by Percoll gradient centrifugation and the cell purity was determined by 3β-hydroxysteroid dehydrogenase (3β-HSD) staining and a testosterone induction assay. To examine TCDD-induced biological consequences, we measured the changes in the secretion of progesterone and testosterone, as well as transcript levels of some selected steroidogenic enzymes (i.e. StAR, P450scc, 3β-HSD and CYP17α), in TCDD/human chorionic gonadotropin (hCG) co-treated cells. Our results indicated that TCDD (0.2 or 2 ng/ml) treatment significantly suppressed hCG (5 or 10 ng/ml)-induced testosterone secretion. The suppressive effect aligned with a reduction of progesterone secretion (P<0.05), as well as a decrease of P450scc mRNA and protein expression (P<0.05). The mechanistic action of TCDD was found to be via the reduction of cellular cAMP levels in the hCG-treated cells. This observation was further confirmed, as the TCDD-mediated suppressive effect could be reversed by dibutyryl cAMP co-treatment. The data indicate that TCDD can modulate cAMP signaling in rat Leydig cells to affect the process of steroidogenesis. © 2005 Society for Endocrinology.link_to_subscribed_fulltex

Changes in endogenous Zn and Cu distribution in different cytosolic protein fractions in mouse liver after administration of a single sublethal dose of CdCl2

The time course of change in tissue Cd, Cu and Zn contents, their distribution in cellular protein fractions as well as the profile of MT gene expression in mouse liver was described over a 7 days period following a single intraperitoneal injection of 2 mg/kg of CdCl2. The result showed that Cd accumulated rapidly in mouse liver. Between 1 h and 7 days after administration, over 18% of the total Cd administered were found in the liver. Cd administration was also associated with the overexpression of the MT-mRNA. However, the time course of induction was not parallel to the change in tissue Cd content. When separated on a Sephadex G-75 column, majority of Cd was found to bind to the fractions known to contain the metal-binding protein, metallothionein (MT). From day 2 after Cd administration, a small amount of the metal was also found associated with the high molecular weight (HMW) proteins. In addition to Cd, tissue Zn content was affected most during the entire study. There was a significant decrease in tissue Zn content during the initial 8 h but tissue Zn content increased significantly throughout the following 6 days. At 1-7 days, majority of Zn was associated with the HMW protein fraction. Although there was no significant change in total tissue Cu content, distribution of Cu in different protein fractions was detected. While in control aniamls, Cu was mainly associated with the HMW proteins, some was found in the MT fraction on the second day. On the 7th day, Cu distribution had deteriorated. Together with changes seen in Cd, the results might suggest that injury had occurred in the tissue at this time. The results of the present study showed that Cd caused a change in subcellular distribution of tissue endogenous metals, which might reflect alteration of cellular functional activities. (C) 2000 Elsevier Science Ireland Ltd.link_to_subscribed_fulltex