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Acinic cell carcinoma (AciCC) may pose a diagnostic challenge, particularly on small biopsies and fine needle aspiration (FNA) because of its variable histology including potential high-grade transformation and its mimickers. Immunoreactivity with circumferential membranous staining for DOG1 can support the diagnosis of AciCC but is not entirely specific. A novel rearrangement t(4;9)(q13;q31) leading to up-regulation of nuclear receptor subfamily 4 group A member 3 (NR4A3) has been described in AciCC, is potentially detectable by fluorescence in situ hybridization (FISH) and may be useful in the evaluation for AciCC. Using NR4A3 Dual Color Break Apart Probe (ZytoVision, Germany) FISH was performed on AciCCs from 3 large academic institutions. NR4A3 rearrangement was defined as positive signal patterns in 15% of tissue interphase nuclei. Fifty-two AciCCs including 47 resections and 5 FNAs (including 5 paired FNA/resections) were analyzed. Five non-AciCC salivary gland tumors and 2 sialadenitis cases were used as controls. Eight AciCCs (15%; 8/52) failed FISH testing. FISH was positive in 23 AciCCs (sensitivity 59%, 23/39) with 100% concordance between 5 matched resection/FNAs (3 were positive for FISH and 2 were negative). FISH was negative in all non-AciCCs (specificity: 100%, 0/7). NR4A3 FISH has a sensitivity of 59% and specificity of 100% in detecting AciCC, which suggests that NR4A3 rearrangement-driven up-regulation is a recurrent, specific oncogenic event in AciCC, consistent with prior results. Hundred percent concordance between matched FNA/resection samples validates its potential utility on cytology samples.

Hydrogen isotope separation is a challenging task due to their similar properties. Herein, based on the chemical affinity quantum sieve (CAQS) effect, the D2/H2 separation performance of M2(m-dobdc) (M = Co, Ni, Mg, Mn; m-dobdc4− = 4,6-dioxido-1,3-benzenedicarboxylate), a series of honeycomb-shaped MOFs with high stability and abundant open metal sites, are studied by gases sorption and breakthrough experiments, in which two critical factors, gas uptake and adsorption enthalpy, are taken into consideration. Among these MOFs, Co2(m-dobdc) exhibits the longest D2 retention time of 180 min/g (H2/D2/Ne: 1/1/98) at 77 K because of its second-highest adsorption enthalpy (10.7 kJ/mol for H2 and 11.8 kJ/mol for D2) and the best sorption capacity (5.22 mmol/g for H2 and 5.49 mmol/g for D2) under low pressure of 1 kPa and 77 K), which make it a promising material for industrial hydrogen isotope separation. Moreover, the results indicate that H2 and D2 capacities under low pressure (about 1 kPa) dominate the final D2/H2 separation property of MOFs.

In various cancers, microRNAs (miRNAs) are abnormally expressed, including thyroid cancer (TC). In recent years, the incidence of TC has increased annually around the world. Compared with female patients, male TC patients are more likely to have a postoperative recurrence and lymph node metastasis, and hence need second treatments. However, the molecular biological processes underlying this phenomenon are not understood. Therefore, we collected data on miRNA expression and clinical information of male TC patients from The Cancer Genome Atlas (TCGA) database. Differentially expressed miRNAs were identified between male TC tissues and matched normal tissues. The Kaplan–Meier method, univariate and multivariate Cox regressions, and receiver operating characteristic curve analyses were performed to assess the association between miRNAs and the disease-free survival of male TC patients. Gene Ontology (GO) and the Kyoto Encyclopaedia of Gene and Genome (KEGG) enrichment analyses were then used to explore the function of miRNA target genes. Furthermore, we evaluated the ability of the miRNA biomarker to predict survival in female TC patients. As a result, a total of 118 differentially expressed miRNAs were identified, including 25 upregulated and 93 downregulated miRNAs. Among them, miR-451a and miR-16-1-3p were confirmed to be independent prognostic factors for the disease-free survival rate. The target genes of miR-451a and miR-16-1-3p were identified, and functional analysis showed that these genes were enriched in 25 Go and KEGG accessions, including cell signal transduction, motor adhesion, phagocytosis, regulation of transcription, cell proliferation, angiogenesis, etc. Neither miR-451a and miR-16-1-3p, nor a prediction model based on both miRNAs effectively predicted survival in female TC patients. In conclusion, both miR-451a and miR-16-1-3p may play important roles in the processes of male TC. The two-miRNA signature involving miR-1258 and miR-193a may serve as a novel prognostic biomarker for male TC patients.

This study investigated the effects of dicyandiamide, phosphogypsum and superphosphate on greenhouse gas emissions and compost maturity during pig manure composting. The results indicated that the addition of dicyandiamide and phosphorus additives had no negative effect on organic matter degradation, and could improve the compost maturity. Adding dicyandiamide alone reduced the emissions of ammonia (NH

The high antibiotic resistance gene (ARGs) contents in livestock manure pose a potential risk to environment and human health. The heap composting with an ambient temperature and thermophilic composting are two methods for converting livestock manure into fertilizer. This study investigated the variations in ARGs and mobile genetic elements (MGEs) and revealed potential mechanisms for ARGs removal using the two composting methods. The ARGs abundance were enriched by 44-fold in heap composting, among them, the macrolide-resistance genes increased significantly. On the contrary, the ARGs were removed by 92% in thermophilic composting, among them, tetracycline-resistance genes decreased by 97%. The bacterial hosts of ARGs were associated with the variations of ARGs and MGEs. The tetO was correlated with the most diverse bacteria in heap composting, and Bacteroidetes was the major host bacteria. While tetT was correlated with the most diverse bacteria in thermophilic composting, and Proteobacteria was the major host bacteria. Structural equation models showed that the enrichment of ARGs in heap composting was mainly correlated with bacterial communities, whereas, the removal of ARGs in thermophilic composting was directly affect by MGEs. Composting temperature directly affected the variations in ARGs. Higher and lower temperatures significantly decreased and increased, respectively, ARGs and MGEs abundance levels.

BackgroundThyroid carcinoma (TC) is the most common malignant endocrine tumor worldwide. Several studies have documented that male patients with TC have a higher rate of metastasis and disease recurrence than female patients. However, the mechanism underlying this observation is not completely clear. The goal of our research was to investigate the potential key candidate genes and pathways related to TC progression in male patients at the molecular level.MethodsA total of 320 samples were obtained from The Cancer Genome Atlas (TCGA) and Genotype-Tissue Expression (GTEx) databases. Hub genes were screened out using weighted gene coexpression network analysis (WGCNA) and a protein–protein interaction (PPI) network analysis. Survival analysis was used to identify hub genes associated with disease-free survival (DFS) rates. Estimation of STromal and Immune cells in MAlignant Tumor tissues using Expression (ESTIMATE) data were used to assess the relationship between hub genes and immune cell infiltration. The molecular mechanism and biological functions of hub genes were explored using RT-qPCR, Western blot, Cell Counting Kit-8 Assay, flow cytometry, Transwell assays, and scratch assays.ResultsForty-seven hub genes were identified, and the survival analysis demonstrated that anti-silencing function 1B (ASF1B) was the sole independent risk factor for poor DFS in male TC patients. Possible associations between the results from the ESTIMATE analysis showed that the ASF1B expression level was related to the ESTIMATE score, immune score, and T-cell regulatory (Treg) infiltration level. Through in vitro cell function experiments, we verified that knockdown of ASF1B inhibited KTC-1 cell proliferation, promoted cell apoptosis, and blocked cell cycle. The silencing of ASF1B reduced protein kinase B (AKT), phospho-AKT (p-AKT), and forkhead box p3 (FOXP3) in KTC-1 cells. Moreover, FOXP3 overexpression markedly restored the cell migration, invasion, and proliferation abilities repressed by ASF1B knockdown.ConclusionsOur results indicate that ASF1B can be considered a prognostic marker, therapeutic target, and predictor of immunotherapy response in male thyroid cancer patients. However, further in-depth studies are required to validate this finding.

Background Granulomatous inflammation is considered an immune mechanism against infection or certain non-neoplastic conditions. Sarcoidosis-like granuloma reactions (SGRs) in primary tumors or in regional lymph nodes are occasionally observed; however, they are rare in lung cancer. Case review In this study, we reported on a case of squamous lung cancer with swollen mediastinal lymph nodes, similar to sarcoidosis. He was misdiagnosed as benign lymph node SGRs or tuberculosis for 2 years. Reexamination of the lymph node by immunohistochemistry confirmed the malignant disease. The lung cancer appeared to remain in the stasis phase for 2 years and then burst to stage IV with the amplification of the fibroblast growth factor receptor 1 gene. Conclusions Although tumor-induced draining lymph node granulomatous reactions are rare, they did exist in some of the patients. In this case, differential diagnoses with malignant granulomas should be performed carefully to avoid the misdiagnosis of a benign disease. The biological significance of such a granulomatous response in inducing tumor remission or in shielding tumor cells from host lymphocytes remains obscure.

Three peritrichous ciliates, Zoothamnium arcuatum n. sp., Z. grossi n. sp., and Z. parahentscheli Sun et al., 2009, were collected from an estuary of the Taehwagang River, Korea. All these species were investigated based on live observations and silver staining, and their small subunit (SSU) rRNA gene was also sequenced. Zoothamnium arcuatum can be identified by a goblet-shaped colony, double-layered peristomial lip, and abstomally shortened row 3 of infundibular polykinety 3 (P3). Zoothamnium grossi is morphologically characterized by an alternately branched stalk with the lowest secondary stalk diverging from the main part of colony, asymmetrically bell-shaped zooids, and three short, parallel ciliary rows in P3. Phylogenetic analysis revealed that the three Zoothamnium species described in this paper clustered with other members of the family Zoothamniidae, as expected.

A hypotrichous ciliate, Paracladotricha salina n. g., n. sp., was discovered in hypersaline waters (salinity about 80‰) from Qingdao, China. Its morphology and some major ontogenetic stages were studied and the phylogenetic position was estimated using standard methods. Paracladotricha salina is characterized by a flexible, more or less slender body (size 50–120 × 20–35 μm), a gonostomatid oral apparatus, one short and two long frontoventral rows, four macronuclear nodules, almost completely reduced dorsal kineties 1–3, and a loss of several parts of the ciliature, namely, the slightly shortened ciliary row of the adoral membranelles, the paroral, and the buccal, the postoral and pretransverse ventral, the transverse, and the caudal cirri. The ontogenesis is rather simple: anlage II of both filial products and anlage III of the opisthe originate de novo, while anlagen IV and V are formed within the parental rows. This combination of features requires the establishment of a new genus, Paracladotricha, which is, according to the morphological data, closely related to Schmidingerothrix and Cladotricha. The small-subunit rRNA gene was sequenced, indicating that P. salina is, as also demonstrated by the oral apparatus, a member of the gonostomatids. We provide a first, vague hypothesis about the phylogenetic relationships of the Gonostomatidae, Cladotrichidae, and Schmidingerotrichidae. However, since molecular data of the type species of these higher taxa are lacking, their validity and relationships remain obscure.

A Gram-negative, aerobic, copper-resistant bacterium, designated strain CU4T, was isolated from copper mine soil in Daye, China. Phylogenetic analysis based on 16S rRNA gene sequences showed highest similarity to Sphingobium rhizovicinum CC-FH12-1T (98.4 %), followed by Sphingobium francense Sp+T (97.2 %), Sphingobium japonicum UT26T (97.1 %), Sphingobium abikonense NBRC 16140T (97.0 %), Sphingobium xenophagum DSM 6383T (96.9 %) and Sphingobium yanoikuyae DSM 7462T (95.5 %). The major fatty acids (>5 %) were summed feature 7 (C18 : 1ω7c, C18 : 1ω9t and/or C18 : 1ω12t), summed feature 4 (C16 : 1ω7c and/or iso-C15 : 0 2-OH), C16 : 0 and C14 : 0 2-OH, and the predominant quinone was ubiquinone Q-10. Spermidine was the major polyamine component. The major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, sphingoglycolipid, phosphatidyldimethylethanolamine and phosphatidylcholine. The genomic DNA G+C content of strain CU4T was 64.9 mol%. Comparison of DNA–DNA hybridization, phenotypic and chemotaxonomic characteristics between strain CU4T and phylogenetically related strains revealed that the new isolate represents a novel species of the genus Sphingobium , for which the name Sphingobium cupriresistens sp. nov. is proposed. The type strain is CU4T ( = KCTC 23865T = CCTCC AB 2011146T). An emended description of the genus Sphingobium is also proposed.

This study investigates the effectiveness and mechanism of decreasing the bioavailability of Pb in bacterial culture and in pot experiments of Pb-contaminated paddy soil by Alishewanella sp. WH16-1. The WH16-1 strain was isolated from mine soil and exhibited high resistances to many heavy metals, especially to Pb2+ (2070 mg/L) and Cr (VI) (2340 mg/L). During cultivation of the WH16-1 strain with the addition of 100 mg/L Pb2+, Pb2+ was precipitated, and 84.13 % of Pb2+ was removed in 72 h. The precipitant was observed by transmission electron microscopy (TEM) and further confirmed to be PbS by X-ray diffraction (XRD) and X-ray photoelectron spectroscopy (XPS). The WH16-1 strain was incubated in Pb2+-added paddy soil pot experiments for 60 days and compared with the uninoculated Pb2+-added paddy soil. Comparison showed that the exchangeable and carbonate-bound Pb in the paddy soil decreased by 14.04 and 10.69 % (P

Alishewanella sp. WH16-1 (= CCTCC M201507) is a facultative anaerobic, motile, Gram-negative, rod-shaped bacterium isolated from soil of a copper and iron mine. This strain efficiently reduces chromate (Cr6+) to the much less toxic Cr3+. In addition, it reduces sulfate (SO42−) to S2−. The S2− could react with Cd2+ to generate precipitated CdS. Thus, strain WH16-1 shows a great potential to bioremediate Cr and Cd contaimination. Here we describe the features of this organism, together with the draft genome and comparative genomic results among strain WH16-1 and other Alishewanella strains. The genome comprises 3,488,867 bp, 50.4 % G + C content, 3,132 protein-coding genes and 80 RNA genes. Both putative chromate- and sulfate-reducing genes are identified. Electronic supplementary material The online version of this article (doi:10.1186/s40793-016-0169-3) contains supplementary material, which is available to authorized users.

isolated from mussel-farming waters near Qingdao, China, are described based on living and protargol-impregnated specimens. Morphologically, the new species can be distinguished from its known congeners by itselongate body shape, narrow oral field, having fewer dorsal kineties and caudal cirri, more marginal cirri, anddifferentiated pretransverse cirri. The identification as a new species is firmly supported by the sequences of thesmall subunit ribosomal rRNA (SSU rRNA) gene, compared with other known

The systematic position of the scuticociliate, Philaster, has been highly ambiguous owing to a lack of either morphogenetic or molecular information. The small subunit rRNA gene of a new species, Philaster apodigitiformis sp. nov. was isolated from eggs of the puffer fish (Takifugu rubripes), was sequenced, and phylo‐genetic trees were constructed with different methods to assess its position and relationship among the scuticociliates. Results indicate that (1) Philaster apodigitiformis clearly falls into the order Philasterida and it is most closely related to the Uronema‐Paranophrys assemblage, which groups with clusters consisting of families Uronematidae, Parauronematidae, Thyrophilacidae and Entorhipidiidae; (2) the two genera Philaster and Philasterides presently assigned to the family Philaster‐idae are in widely separated clades, indicating that the family is polyphyletic; and (3) the morphological similarity of Philaster and Philasterides is probably an example of evolutionary convergenc...

A population of Kiitricha minuta Xu et al., 2008, a small kiitrichid ciliate, was isolated from a brackish water sample in Jiaozhou Bay, Qingdao, northern China. After comparison of its morphology and infraciliature, it is believed that this morphotype should be assigned to the genus Caryotricha; hence, a new combination is suggested, Caryotricha minuta (Xu et al., 2008) nov. comb. The small-subunit (SSU) rRNA gene sequence was determined in order to elucidate the phylogenetic position of this poorly known, ambiguous genus. The organism can be clearly separated from its congener, Caryotricha convexa Kahl, 1932, by the extremely shortened ventral cirral rows in the posterior ends. Based on the data available, an improved diagnosis is given for the genus: marine Kiitrichidae with prominent buccal field; two highly developed undulating membranes; non-grouped, uniform cirral rows on both ventral and dorsal sides; enlarged transverse cirri present, which are the only differentiated cirri; marginal cirri not present; one short migratory row located posterior to buccal field; structure of dorsal kineties generally in Kiitricha pattern. The sequence of the SSU rRNA gene of C. minuta differs by 13 % from that of Kiitricha marina. Molecular phylogenetic analyses (Bayesian inference, least squares, neighbour joining, maximum parsimony) indicate that Caryotricha, together with Kiitricha, diverges at a deep level from all other spirotrichs. Its branching position is between Phacodiniidia and Licnophoridia. The results strongly support the distinct separation of the Kiitricha-Caryotricha clade, which always branches basal to the Stichotrichia-Hypotrichia-Oligotrichia-Choreotrichia assemblage. These results also confirm the previous hypothesis that the Kiitricha-Caryotricha group, long assumed to be a close relation to the euplotids, represents a taxon at subclass level within the spirotrichs.

The morphology, morphogenesis and small subunit rRNA (SSrRNA) gene sequence of a new marine euplotid ciliate, Diophrys apoligothrix spec. nov., isolated from a sandy beach near Qingdao, north China, were investigated. This organism is characterized as follows: rigid body with a sculptured surface; size in vivo about 60-110 x 40-70microm; adoral zone comprising about 28 membranelles; five frontal, two ventral, five transverse, one left marginal and three caudal cirri; five dorsal kineties with sparsely distributed dikinetids; two ellipsoid macronuclear nodules; marine habitat. Although the main morphogenetic events are similar to those reported in other congeners, three features are noteworthy: (1) the parental adoral zone is retained almost unchanged by the proter except for the outer margin of membranelles in the proximal portion, where the de-differentiation and then reorganization take place in situ in the mid-to-late phases of division; (2) the dorsal kinety anlagen are formed via a two-step mode; (3) the marginal cirrus anlage is formed de novo near the old structure and does not divide. Based on the morphological, morphogenetic and SSrRNA gene sequence data, it was concluded that D. apoligothrix is a well-outlined and distinctive member of the genus Diophrys.

Figure S1. Phylogenetic relationships of Alishewanella sp. WH16-1 based on gyrB sequences. The analysis was performed by MEGA 6.0 [45] with NJ algorithm and 1,000 bootstrap repetitions were computed to estimate the reliability of the tree. The gyrB gene of strain WH16-1 is the gene sequence coding for AAY72_13600. Figure S2. The putative sulfate transport and reduction pathway in Alishewanella sp. WH16-1. APS stands for adenylylsulphate, PAPS stands for phosphoadenylylsulphate. The locus tag numbers of the predicted proteins (CysP, CysU, CysW, CysA, CysN, CysD, CysC, CysH CysJ and CysI) are AAY72_14890, AAY72_14885, AAY72_14880, AAY72_14875, AAY72_03865, AAY72_03870, AAY72_07290, AAY72_07265, AAY72_07255 and AAY72_07260, respectively. (DOCX 355Â kb)

Two insufficiently described marine euplotids, Diophrys irmgard Mansfeld (1923) and Diophrys hystrix Buddenbrock (1920), found in coastal waters near Qingdao, China, were reinvestigated with emphasis on their living morphology and infraciliature. Improved diagnoses and redescriptions are supplied for both organisms. Critical redefinitions of nine morphotypes are presented and the validities of the nominal species are discussed and briefly revised. Following Curds and Wu (1983), and based on data obtained during the present study and a critical review of the literature, five additional nominal species are regarded as junior synonyms of known forms: Diophrys quadricaudatus Agamaliev (1967), D. scutoides Agamaliev (1967), and D. tetramacronucleata Kattar (1970) are synonymized with D. scutum (Dujardin 1841), while D. pentacirratus Alekperov (1984) and D. multicirratus Alekperov (1984) are junior synonyms of D. oligothrix Borror (1965).

Amplification/duplication of the BCR-ABL gene has been found to be one of the key factors leading to drug resistance to imatinib mesylate (IM). In the present study, we used G-banding to identify the presence of de novo identical isodicentric chromosomes in IM-resistant patients. Fluorescence in situ hybridization (FISH) analysis on interphase nuclei confirmed the heterogeneity and amplification of the fused BCR-ABL gene. FISH analysis superimposed on G-banding confirmed the presence of isodicentric Philadelphia chromosomes. The impact of the isodicentric Philadelphia chromosomes on genomic instability, heterogeneity, and amplification of the Philadelphia chromosomes in IM-resistant patients is discussed.

The morphology and taxonomy of two marine Amphisiella species, isolated from mariculture waters in northern China, were investigated using standard techniques. One species corresponds well with the original description of the poorly known Amphisiella milnei (Kahl, 1932) Horvath, 1950 by remarkable characteristics, inter alia, (i) the additional cirri between the left frontal cirrus and the buccal cirrus, and (ii) ring-shaped structure in the anterior and posterior body portion (posterior one sometimes lacking). The detailed description of the cirral pattern and an informative ontogenetic stage first reveal that the additional cirri, which are uniformly absent in all congeners, originate from the frontal-ventral transverse cirral anlage I. The other species represents a new species of Amphisiella, A. sinica sp. nov. It is distinguished from its most closely related congener A. annulata (Kahl, 1928) Borror, 1972 by having numerous ring-shaped structures with a shallow brim more or less densely centralised at both ends of the body. Brief notes on the cell division and phylogenetic analyses based on small subunit (SSU) rRNA gene sequence for both organisms are also supplied in order to get further understanding of their systematic positions. The molecular information indicates that both organisms belong to two separate clades and confirms that the genus Amphisiella might be polyphyletic.

Eosinophilic/T-cell chorionic vasculitis (ETCV) is an inflammatory lesion of placental fetal vessels. In contrast to acute chorionic vasculitis, inflammation in ETCV is seen in chorionic vessel walls opposite the amnionic surface. It is not known whether inflammation in ETCV consists of maternal cells from the intervillous space or fetal cells migrating from the vessel. We used fluorescent in situ hybridization (FISH) to differentiate fetal versus maternal cells in ETCV. Placentas with ETCV, previously identified for a published study, were used. Infant sex in each case was identified using the electronic medical record. For male infants, 3-μm sections were cut from archived tissue blocks from placentas involving ETCV and stained with fluorescent X- and Y-chromosome centromeric probes. A consecutive hematoxylin/eosin-stained section was used for correlation. FISH analysis was performed on 400 interphase nuclei at the site of ETCV to determine the proportion of XX, XY, X, and Y cells. Of 31 ETCV cases, 20 were female and 10 were male (1 sex not recorded). Six of 10 cases with male infants had recuts with visible ETCV. In these 6 cases the average percentages (ranges) of XY cells, X-only cells, and Y-only cells in the region of inflammation were 81 (70–90), 11 (6–17), and 8 (2–14), respectively. There was a 2:1 female:male infant ratio in ETCV. Similar to acute chorionic vasculitis, the inflammation in ETCV is of fetal origin. It is still unknown, however, whether the stimulus for ETCV is of fetal or maternal origin.

Granulomatous inflammation is considered an immune mechanism against infection or certain non-neoplastic conditions. Sarcoidosis-like granuloma reactions (SGRs) in primary tumors or in regional lymph nodes are occasionally observed; however, they are rare in lung cancer.In this study, we reported on a case of squamous lung cancer with swollen mediastinal lymph nodes, similar to sarcoidosis. He was misdiagnosed as benign lymph node SGRs or tuberculosis for 2 years. Reexamination of the lymph node by immunohistochemistry confirmed the malignant disease. The lung cancer appeared to remain in the stasis phase for 2 years and then burst to stage IV with the amplification of the fibroblast growth factor receptor 1 gene.Although tumor-induced draining lymph node granulomatous reactions are rare, they did exist in some of the patients. In this case, differential diagnoses with malignant granulomas should be performed carefully to avoid the misdiagnosis of a benign disease. The biological significance of such a granulomatous response in inducing tumor remission or in shielding tumor cells from host lymphocytes remains obscure.

Uptake of organic solutes and xenobiotics by mammalian cells is mediated by ATP-independent transporters, and four families of transporters have now been identified. To search for novel organic solute transporters, a liver cDNA library from an evolutionarily primitive marine vertebrate, the little skate Raja erinacea , was screened for taurocholate transport activity by using Xenopus laevis oocytes. In contrast to the organic anion transporters identified to date, a transport activity was identified in this library that required the coexpression of two distinct gene products, termed organic solute transporter α and β (Ostα, Ostβ). Ostα cDNA encodes for a protein of 352 aa and seven putative transmembrane (TM) domains. Ostβ contains 182 aa and has at least one and perhaps two TM domains. There is no significant sequence identity between Ostα and Ostβ, and only low identity with sequences in the databases; however, Ostα bears a resemblance to some G protein-coupled receptors, and Ostβ exhibits 22% amino acid identity with the C-terminal TM and intracellular domains of protocadherin-γ, a cell surface glycoprotein. Xenopus oocytes injected with the cRNA for both Ostα and Ostβ, but not each separately, were able to take up taurocholate, estrone sulfate, digoxin, and prostaglandin E 2 , but not p -aminohippurate or S -dinitrophenyl glutathione. Transport was sodium-independent, saturable, and inhibited by organic anions and steroids, including the major skate bile salt, scymnol sulfate. These results identify an organic anion transporter composed of a putative seven-helix TM protein and an ancillary membrane polypeptide.

One member of the OATP family of transporters, rat Oatp1, functions as an anion exchanger that is driven in part by the glutathione (GSH) electrochemical gradient, indicating that other OATP-related transporters may also be energized by this mechanism. The present study examined whether rat Oatp2 is also an anion exchanger, and, if so, whether it is energized by the GSH electrochemical gradient. As with Oatp1, uptake of 10 microM [(3)H]taurocholate in Oatp2-expressing Xenopus laevis oocytes was trans-stimulated by intracellular 0.2 mM unlabeled taurocholate, indicating bidirectional transport. Interestingly, [(3)H]taurocholate uptake in Oatp2-expressing oocytes was also trans-stimulated when oocytes were preloaded with GSH, S-methylglutathione, S-sulfobromophthalein-glutathione, S-dinitrophenyl glutathione, or ophthalmic acid (a GSH analog) but not by glutarate or N-acetylcysteine, suggesting that GSH derivatives and conjugates may function as intracellular substrates for Oatp2. Support for this hypothesis was provided by the demonstration of enhanced [(3)H]GSH and [(3)H]S-(2,4-dinitrophenyl)-glutathione efflux in Oatp2-expressing oocytes. However, in contrast to Oatp1, extracellular GSH failed to cis-inhibit uptake of [(3)H]taurocholate or [(3)H]digoxin in Oatp2-expressing oocytes, indicating that the stimulatory effect of high intracellular GSH concentrations is not due to a coupled exchange mechanism. Taken together, the results indicate that Oatp2 mediates bidirectional transport of organic anions by a GSH-sensitive facilitative diffusion mechanism and suggest that this transporter may play a role in cellular export of specific organic molecules.

A hypotrichous ciliate, Paracladotricha salina n. g., n. sp., was discovered in hypersaline waters (salinity about 80‰) from Qingdao, China. Its morphology and some major ontogenetic stages were studied and the phylogenetic position was estimated using standard methods. Paracladotricha salina is characterized by a flexible, more or less slender body (size 50-120 × 20-35 μm), a gonostomatid oral apparatus, one short and two long frontoventral rows, four macronuclear nodules, almost completely reduced dorsal kineties 1-3, and a loss of several parts of the ciliature, namely, the slightly shortened ciliary row of the adoral membranelles, the paroral, and the buccal, the postoral and pretransverse ventral, the transverse, and the caudal cirri. The ontogenesis is rather simple: anlage II of both filial products and anlage III of the opisthe originate de novo, while anlagen IV and V are formed within the parental rows. This combination of features requires the establishment of a new genus, Paracladotricha, which is, according to the morphological data, closely related to Schmidingerothrix and Cladotricha. The small-subunit rRNA gene was sequenced, indicating that P. salina is, as also demonstrated by the oral apparatus, a member of the gonostomatids. We provide a first, vague hypothesis about the phylogenetic relationships of the Gonostomatidae, Cladotrichidae, and Schmidingerotrichidae. However, since molecular data of the type species of these higher taxa are lacking, their validity and relationships remain obscure.