bluuebunny/crossref_metadata_2025 huggingface dataset:
The two C-terminal tyrosines stabilize occluded Na/K pump conformations containing Na or K ions
Authors: Vedovato Natascia , Gadsby David C.
Published: June 2010

Interactions of the three transported Na ions with the Na/K pump remain incompletely understood. Na/K pump crystal structures show that the extended C terminus of the Na,K–adenosine triphosphatase (ATPase) α subunit directly contacts transmembrane helices. Deletion of the last five residues (KETYY in almost all Na/K pumps) markedly lowered the apparent affinity for Na activation of pump phosphorylation from ATP, a reflection of cytoplasmic Na affinity for forming the occluded E1P(Na3) conformation. ATPase assays further suggested that C-terminal truncations also interfere with low affinity Na interactions, which are attributable to extracellular effects. Because extracellular Na ions traverse part of the membrane’s electric field to reach their binding sites in the Na/K pump, their movements generate currents that can be monitored with high resolution. We report here electrical measurements to examine how Na/K pump interactions with extracellular Na ions are influenced by C-terminal truncations. We deleted the last two (YY) or five (KESYY) residues in Xenopus laevis α1 Na/K pumps made ouabain resistant by either of two kinds of point mutations and measured their currents as 10-mM ouabain–sensitive currents in Xenopus oocytes after silencing endogenous Xenopus Na/K pumps with 1 µM ouabain. We found the low affinity inhibitory influence of extracellular Na on outward Na/K pump current at negative voltages to be impaired in all of the C-terminally truncated pumps. Correspondingly, voltage jump–induced transient charge movements that reflect pump interactions with extracellular Na ions were strongly shifted to more negative potentials; this signals a several-fold reduction of the apparent affinity for extracellular Na in the truncated pumps. Parallel lowering of Na affinity on both sides of the membrane argues that the C-terminal contacts provide important stabilization of the occluded E1P(Na3) conformation, regardless of the route of Na ion entry into the binding pocket. Gating measurements of palytoxin-opened Na/K pump channels additionally imply that the C-terminal contacts also help stabilize pump conformations with occluded K ions.
DARNED: a DAtabase of RNa EDiting in humans
Authors: Kiran Anmol , Baranov Pavel V.
Published: June 2010

Motivation: RNA editing is a phenomenon, which is responsible for the alteration of particular nucleotides in RNA sequences relative to their genomic templates. Recently, a large number of RNA editing instances in humans have been identified using bioinformatic screens and high-throughput experimental investigations utilizing next-generation sequencing technologies. However, the available data on RNA editing are not uniform and difficult to access.

Results: Here, we describe a new database DARNED (DAtabase of RNa EDiting) that provides centralized access to available published data related to RNA editing. RNA editing locations are mapped on the reference human genome. The current release of the database contains information on approximately 42 000 human genome coordinates corresponding to RNA locations that undergo RNA editing, mostly involving adenosine-to-inosine (A-to-I) substitutions. The data can be queried using a range of genomic coordinates, their corresponding functional localization in RNA molecules [Exons, Introns, CoDing Sequence (CDS) and UnTranslated Regions (UTRs)] and information regarding tissue/organ/cell sources where RNA editing has been observed. It is also possible to obtain RNA editing information for a specific gene or an RNA molecule using corresponding accession numbers. Search results provide information on the number of expressed sequence tags (ESTs) supporting edited and genomic bases, functional localization of RNA editing and existence of known single nucleotide polymorphisms (SNPs). Editing data can be explored in UCSC and Ensembl genome browsers, in conjunction with additional data provided by these popular genome browsers. DARNED has been designed for researchers seeking information on RNA editing and for the developers of novel algorithms for its prediction.

Availability: DARNED is accessible at http://darned.ucc.ie

Contact:  p.baranov@ucc.ie; brave.oval.pan@gmail.com
Electroacupuncture at the Zusanli (ST‐36) Acupoint Induces a Hypoglycemic Effect by Stimulating the Cholinergic Nerve in a Rat Model of Streptozotocine‐Induced Insulin‐Dependent Diabetes Mellitus
Authors: Lee Yu-Chen , Li Te-Mao , Tzeng Chung-Yuh , Chen Ying-I , Ho Wai-Jane , Lin Jaung-Geng , Chang Shih-Liang
Published: June 2010

Animal studies have shown that electroacupuncture (EA) at Zusanli (ST‐36) and Zhongwan (CV‐12) acupoints reduces plasma glucose concentrations in rats with type II diabetes. However, whether EA reduces plasma glucose levels in type I diabetes is still unknown. In this study, we explore the various non‐insulin‐dependent pathways involved in EA‐induced lowering of plasma glucose. Streptozotocin (STZ) (60 mg kg−1, i.v.) was administered via the femoral vein to induce insulin‐dependent diabetes in non‐adrenalectomized and in adrenalectomomized rats. EA (15 Hz) was applied for 30 min to bilateral ST‐36 acupoints after administration of Atropine (0.1 mg kg−1 i.p.), Eserine (0.01 mg kg−1 i.p.), or Hemicholinium‐3 (5 μg kg−1 i.p.) in non‐adrenalectomized rats. Rats administered acetylcholine (0.01 mg kg−1 i.v.) did not undergo EA. Adrenalectomized rats underwent EA at bilateral ST‐36 acupoints without further treatment. Blood samples were drawn from all rats before and after EA to measure changes in plasma glucose levels. Expression of insulin signaling proteins (IRS1, AKT2) in atropine‐exposed rats before and after EA was measured by western blot. Atropine and hemicholinium‐3 completely blocked the plasma glucose lowering effects of EA, whereas eserine led to a significant hypoglycemic response. In addition, plasma glucose levels after administration of acetylcholine were significantly lower than the fasting glucose levels. In STZ‐adrenalectomized rats, EA did not induce a hypoglycemic response. EA stimulated the expression of IRS1 and AKT2 and atropine treatment blocked the EA‐induced expression of those insulin signaling proteins. Taken together, EA at the ST‐36 acupoint reduces plasma glucose concentrations by stimulating the cholinergic nerves.
Genetic analysis of sleep
Authors: Crocker Amanda , Sehgal Amita
Published: June 2010

Almost 20 years ago, the gene underlying fatal familial insomnia was discovered, and first suggested the concept that a single gene can regulate sleep. In the two decades since, there have been many advances in the field of behavioral genetics, but it is only in the past 10 years that the genetic analysis of sleep has emerged as an important discipline. Major findings include the discovery of a single gene underlying the sleep disorder narcolepsy, and identification of loci that make quantitative contributions to sleep characteristics. The sleep field has also expanded its focus from mammalian model organisms to Drosophila, zebrafish, and worms, which is allowing the application of novel genetic approaches. Researchers have undertaken large-scale screens to identify new genes that regulate sleep, and are also probing questions of sleep circuitry and sleep function on a molecular level. As genetic tools continue to be refined in each model organism, the genes that support a specific function in sleep will become more apparent. Thus, while our understanding of sleep still remains rudimentary, rapid progress is expected from these recently initiated studies.
A novel phosphatase cascade regulates differentiation in Trypanosoma brucei via a glycosomal signaling pathway
Authors: Szöőr Balázs , Ruberto Irene , Burchmore Richard , Matthews Keith R.
Published: June 2010

In the mammalian bloodstream, the sleeping sickness parasite Trypanosoma brucei is held poised for transmission by the activity of a tyrosine phosphatase, TbPTP1. This prevents differentiation of the transmissible “stumpy forms” until entry into the tsetse fly, whereupon TbPTP1 is inactivated and major changes in parasite physiology are initiated to allow colonization of the arthropod vector. Using a substrate-trapping approach, we identified the downstream step in this developmental signaling pathway as a DxDxT phosphatase, TbPIP39, which is activated upon tyrosine phosphorylation, and hence is negatively regulated by TbPTP1. In vitro, TbPIP39 promotes the activity of TbPTP1, thereby reinforcing its own repression, this being alleviated by the trypanosome differentiation triggers citrate and cis-aconitate, generating a potentially bistable regulatory switch. Supporting a role in signal transduction, TbPIP39 becomes rapidly tyrosine-phosphorylated during differentiation, and RNAi-mediated transcript ablation in stumpy forms inhibits parasite development. Interestingly, TbPIP39 localizes in glycosomes, peroxisome-like organelles that compartmentalize the trypanosome glycolytic reactions among other enzymatic activities. Our results invoke a phosphatase signaling cascade in which the developmental signal is trafficked to a unique metabolic organelle in the parasite: the glycosome. This is the first characterized environmental signaling pathway targeted directly to a peroxisome-like organelle in any eukaryotic cell.
Functional conservation of Mei4 for meiotic DNA double-strand break formation from yeasts to mice
Authors: Kumar Rajeev , Bourbon Henri-Marc , de Massy Bernard
Published: June 2010

Meiotic recombination is initiated by the programmed induction of DNA double-strand breaks (DSBs) catalyzed by the evolutionarily conserved Spo11 protein. Studies in yeast have shown that DSB formation requires several other proteins, the role and conservation of which remain unknown. Here we show that two of these Saccharomyces cerevisiae proteins, Mei4 and Rec114, are evolutionarily conserved in most eukaryotes. Mei4−/− mice are deficient in meiotic DSB formation, thus showing the functional conservation of Mei4 in mice. Cytological analyses reveal that, in mice, MEI4 is localized in discrete foci on the axes of meiotic chromosomes that do not overlap with DMC1 and RPA foci. We thus propose that MEI4 acts as a structural component of the DSB machinery that ensures meiotic DSB formation on chromosome axes. We show that mouse MEI4 and REC114 proteins interact directly, and we identify conserved motifs as required for this interaction. Finally, the unexpected, concomitant absence of Mei4 and Rec114, as well as of Mnd1, Hop2, and Dmc1, in some eukaryotic species (particularly Neurospora crassa, Drosophila melanogaster, and Caenorhabditis elegans) suggests the existence of Mei4–Rec114-dependent and Mei4–Rec114-independent mechanisms for DSB formation, and a functional relationship between the chromosome axis and DSB formation.
The interaction between Myc and Miz1 is required to antagonize TGFβ-dependent autocrine signaling during lymphoma formation and maintenance
Authors: van Riggelen Jan , Müller Judith , Otto Tobias , Beuger Vincent , Yetil Alper , Choi Peter S. , Kosan Christian , Möröy Tarik , Felsher Dean W. , Eilers Martin
Published: June 2010

The Myc protein suppresses the transcription of several cyclin-dependent kinase inhibitors (CKIs) via binding to Miz1; whether this interaction is important for Myc's ability to induce or maintain tumorigenesis is not known. Here we show that the oncogenic potential of a point mutant of Myc (MycV394D) that is selectively deficient in binding to Miz1 is greatly attenuated. Binding of Myc to Miz1 is continuously required to repress CKI expression and inhibit accumulation of trimethylated histone H3 at Lys 9 (H3K9triMe), a hallmark of cellular senescence, in T-cell lymphomas. Lymphomas that arise express high amounts of transforming growth factor β-2 (TGFβ-2) and TGFβ-3. Upon Myc suppression, TGFβ signaling is required to induce CKI expression and cellular senescence and suppress tumor recurrence. Binding of Myc to Miz1 is required to antagonize growth suppression and induction of senescence by TGFβ. We demonstrate that, since lymphomas express high levels of TGFβ, they are poised to elicit an autocrine program of senescence upon Myc inactivation, demonstrating that TGFβ is a key factor that establishes oncogene addiction of T-cell lymphomas.
Gamete compatibility between marine and estuarine Acanthopagrus spp. (Sparidae) and their hybrids
Authors: Roberts D. G. , Gray C. A. , West R. J. , Ayre D. J.
Published: June 2010

On Australia's south‐east coast, hybridization between estuary‐restricted black bream Acanthopagrus butcheri Munro and its migratory coastal congener yellowfin bream Acanthopagrus australis (Günther) has led to estuarine populations largely composed of hybrids that are most genetically similar to A. butcheri. The fertilization success achieved when ova of estuary‐caught A. butcheri were fertilized with the cryogenically preserved sperm of either ocean‐caught A. australis or estuary‐caught A. butcheri‐like was compared. The experimental crosses, which by chance included both pure parental and hybrid bream, revealed no evidence that gametic incompatibility provides a barrier to fertilization among both pure species and their hybrids.
Phylogeographical analysis reveals multiple conservation units in brook lampreys Lampetra planeri of Portuguese streams
Authors: Pereira A. M. , Robalo J. I. , Freyhof J. , Maia C. , Fonseca J. P. , Valente A. , Almada V. C.
Published: June 2010

The populations of brook lamprey Lampetra planeri of Portuguese Rivers were analysed phylogeographically using a fragment of 644 bp of the mitochondrial control region of 158 individuals from six populations. Samples representing L. planeri and migratory lampreys Lampetra fluviatilis of rivers draining to the North Sea and the Baltic Sea were also included to assess the relationships of Portuguese samples. The data support a clear differentiation of all the populations studied. Several populations, which are isolated among themselves and also from the migratory lampreys, proved to be entirely composed of private haplotypes, a finding that supports some time of independent evolutionary history for these populations. This, combined with the geographic confinement to small water bodies, justifies the recognition of at least four conservation units in the Portuguese rivers Sado, São Pedro, Nabão and Inha.
Evidence for a common genetic aetiology in high‐risk families with multiple haematological malignancy subtypes
Authors: Tegg Elizabeth M. , Thomson Russell J. , Stankovich Jim , Banks Annette , Flowers Ceri , McWhirter Rebekah , Panton Jean , Piaszczyk Anne , Bahlo Melanie , Marsden Katherine A. , Lowenthal Ray M. , Foote Simon J. , Dickinson Joanne L.
Published: June 2010

A family history of a haematological malignancy (HM) is known to be a risk factor for HMs. However, collections of large families with multiple cases of varied disease types are relatively rare. We describe a collection of 12 families with dense aggregations of multiple HM subtypes. Cases were ascertained from a population based study conducted between 1972 and 1980 in Tasmania, Australia. Diagnoses were confirmed through review and re‐examination of stored tissue, pathology reports, Tasmanian Cancer Registry and flow cytometry records. Family trees were generated and kinship coefficients were calculated for all pairs of affected individuals. 120 cases were found in these families. Cases diagnosed with chronic lymphocytic leukaemia (CLL) demonstrated the most significantly increased aggregation (P < 0·0001). There was also significant evidence that those individuals diagnosed at an older age (>53 years), did not aggregate together in families with disease that presented at an earlier age (<20 years) (P = 0·009).
Degradation of coeliac disease-inducing rye secalin by germinating cereal enzymes: diminishing toxic effects in intestinal epithelial cells
Authors: Stenman S M , Lindfors K , Venäläinen J I , Hautala A , Männistö P T , Garcia-Horsman J A , Kaukovirta-Norja A , Auriola S , Mauriala T , Mäki M , Kaukinen K
Published: June 2010

Currently the only treatment for coeliac disease is a lifelong gluten-free diet excluding food products containing wheat, rye and barley. There is, however, only scarce evidence as to harmful effects of rye in coeliac disease. To confirm the assumption that rye should be excluded from the coeliac patient's diet, we now sought to establish whether rye secalin activates toxic reactions in vitro in intestinal epithelial cell models as extensively as wheat gliadin. Further, we investigated the efficacy of germinating cereal enzymes from oat, wheat and barley to hydrolyse secalin into short fragments and whether secalin-induced harmful effects can be reduced by such pretreatment. In the current study, secalin elicited toxic reactions in intestinal Caco-2 epithelial cells similarly to gliadin: it induced epithelial cell layer permeability, tight junctional protein occludin and ZO-1 distortion and actin reorganization. In high-performance liquid chromatography and mass spectroscopy (HPLC-MS), germinating barley enzymes provided the most efficient degradation of secalin and gliadin peptides and was thus selected for further in vitro analysis. After germinating barley enzyme pretreatment, all toxic reactions induced by secalin were ameliorated. We conclude that germinating enzymes from barley are particularly efficient in the degradation of rye secalin. In future, these enzymes might be utilized as a novel medical treatment for coeliac disease or in food processing in order to develop high-quality coeliac-safe food products.
The effect of Longan seed polyphenols on colorectal carcinoma cells
Authors: Chung Yuan‐Chiang , Lin Chih‐Cheng , Chou Chih‐Chung , Hsu Chih‐Ping
Published: June 2010

Eur J Clin Invest 2010; 40 (8): 713–721

Background  Polyphenol‐rich longan seed extract (LSP) is a free radical scavenger and antioxidant. However, the effect of LSP on the growth of human colorectal carcinoma cells (CRC) has not yet been evaluated.

Materials and methods  Polyphenols of longan seeds were extracted and measured by colorimetry. Four CRC cell lines (Colo 320DM, SW480, HT‐29 and LoVo) were treated with LSP and assessed for viability by trypan blue exclusion, for cell cycle distribution by flow cytometry, for apoptosis by annexin V labelling and for changes in the levels of proteins involved in cell cycle control or apoptosis by immunoblotting.

Results  Total phenol content of LSP was 695 mg g−1 and total flavonoids were 150 mg g−1. LSP inhibited the proliferation (25 μg mL−1–200 μg mL−1) of Colo 320DM, SW480 and HT‐29, but not LoVo. LSP inhibited the proliferation by blocking cell cycle progression during the DNA synthesis phase and inducing apoptotic death. Western blotting indicated that LSP blocks the S phase, reducing the expression of cyclin A and cyclin D1. Colo 320DM and SW480 treated with LSP also showed the activation of caspase 3 and increased Bax : Bcl‐2 ratio.

Conclusion  LSP induces S phase arrest of the cell cycle and apoptotic death in three CRC cell lines. The results indicate that LSP is a potential novel chemoprevention and treatment agent for colorectal cancer.
Potential functional redundancy and resource facilitation between tadpoles and insect grazers in tropical headwater streams
Authors: COLÓN‐GAUD CHECO , WHILES M. R. , BRENES R. , KILHAM S. S. , LIPS K. R. , PRINGLE C. M. , CONNELLY S. , PETERSON S. D.
Published: June 2010

1. We quantified production and consumption of stream‐dwelling tadpoles and insect grazers in a headwater stream in the Panamanian uplands for 2 years to assess their effects on basal resources and energy fluxes. At the onset of our study, this region had healthy, diverse amphibian populations, but a catastrophic disease‐driven decline began in September 2004, which greatly reduced amphibian populations.

2. Insect grazer production was 348 mg ash‐free dry mass (AFDM) m−2 year−1 during the first year of the study and increased slightly to 402 mg AFDM m−2 year−1 during the second year.

3. Prior to amphibian declines, resource consumption by grazers (tadpoles and insects) was estimated at 2.9 g AFDM m−2 year−1 of algal primary production, which was nearly twice the estimated amount available. Insect grazers alone accounted for c. 81% of total primary consumption. During the initial stages of the declines, consumption remained at c. 2.9 g AFDM m−2 year−1, but only 35% of the available resource was being consumed and insect grazers accounted for c. 94% of total consumption.

4. Production and resource consumption of some insect grazers increased during the second year, as tadpoles declined, indicating a potential for functional redundancy in this system. However, other insect grazer taxa declined or did not respond to tadpole losses, suggesting a potential for facilitation between tadpoles and some insects; differential responses among taxa resulted in the lack of a response by insect grazers as a whole.

5. Our results suggest that before massive population declines, tadpoles exerted strong top‐down control on algal production and interacted in a variety of ways with other primary consumers.

6. As amphibian populations continue to decline around the globe, changes in the structure and function of freshwater habitats should be expected. Although our study was focused on tropical headwater streams, our results suggest that these losses of consumer diversity could influence other aquatic systems as well and may even reach to adjacent terrestrial environments.
Are more productive varieties of Paspalum dilatatum less tolerant to drought?
Authors: Couso L. L. , Gatti M. L. , Cornaglia P. S. , Schrauf G. E. , Fernández R. J.
Published: June 2010

Paspalum dilatatum Poir., is a perennial C4 grass widely distributed in the Argentinean Pampas. The response to water availability for materials developed with forage‐production purposes is unknown. We hypothesized that genetic differences between commercial varieties are reflected in their regrowth capacity under water stress. The effect of five levels of constant water supply on three plant varieties (two derived from apomictic materials: ‘Relincho’ and ‘Alonso’ and one from sexually‐derived material: ‘Primo’) were examined in the greenhouse. Leaf‐ and plant‐response traits were followed during 38 d after a single defoliation event. Seven response variables were measured: three of them were morphogenetic (leaf elongation rate, leaf appearance rate and leaf elongation duration) and four were structural (number of live leaves, lamina length, tiller biomass and tiller production). The sexual material showed higher values for growth variables than the apomictic varieties (leaf elongation rate, leaf length and tiller biomass) across the environmental range. Apomictic varieties showed a proportionally similar drought response to the sexual material for the seven variables. No intra‐specific trade‐off (statistical interaction) was found between growth under high water availability conditions and drought tolerance.
Nutritive value of two meadows and relationships with some vegetation traits
Authors: Andueza D. , Cruz P. , Farruggia A. , Baumont R. , Picard F. , Michalet‐Doreau B.
Published: June 2010

This study was designed to compare the evolution in nutritive value between two permanent grasslands that had different botanical composition, during 2 years (2003 and 2004), and to study the relationships between a set of vegetation and digestibility characteristics. Botanical composition, lamina proportions (LP), and in vivo organic matter digestibility (OMD) of grasslands were measured at three different times during the first growth cycle over 2 years: last week in May, third week in June and second week of July. Foliar traits [specific leaf area (SLA) and leaf dry matter content (LDMC)] were measured on grasses in the first period of both years. The LDMC and botanical composition values were used to calculate plant functional types (PFT) of the grasslands at each period. Relationships between OMD and SLA, LDMC, LP and forbs proportions (FP) were tested. Grassland rich in forbs, which was classified in PFT B, showed higher OMD in the first stage of maturity and lower OMD at the end of the cycle in 2004 than grassland rich in grasses, which was classified as PFT C. There were significant relationships between LDMC and OMD across the two first periods. There were also significant relationships between LP and OMD in the first period and between FP and OMD in the third period. The botanical composition of the grassland and consequently its PFT is an important factor explaining the evolution of the nutritive value of permanent grasslands during the first growth cycle.
Decreased production of interleukin‐10 and transforming growth factor‐β in Toll‐like receptor‐activated intestinal B cells in SAMP1/Yit mice
Authors: Mishima Yoshiyuki , Ishihara Shunji , Aziz Md. Monowar , Oka Akihiko , Kusunoki Ryusaku , Otani Aya , Tada Yasumasa , Li Yong‐Yu , Moriyama Ichiro , Oshima Naoki , Yuki Takafumi , Amano Yuji , Matsumoto Satoshi , Kinoshita Yoshikazu
Published: June 2010

A unique subset of B cells expressing interleukin‐10 (IL‐10) and transforming growth factor‐β (TGF‐β) plays an essential role in preventing inflammation and autoimmunity. We investigated the presence of this cell subset in intestines and its role in the pathogenesis of ileitis using SAMP1/Yit and age‐matched control AKR/J mice. Mononuclear cells were isolated from mesenteric lymph nodes (MLNs) and the expressions of B220, CD1d, CD5, Toll‐like receptor 4 (TLR4) and TLR9 in isolated cells were analysed. Purified B cells were stimulated with lipopolysaccharide (LPS) or CpG‐DNA, then IL‐10 and TGF‐β1 expressions were examined by enzyme immunoassay and flow cytometry. Production of IL‐1β by TLR‐mediated macrophages co‐cultured with or without purified MLN B cells from SAMP1/Yit and AKR/J mice was evaluated. In addition, interferon‐γ (IFN‐γ) production in intestinal T cells co‐cultured with MLN B cells were also assessed in SAMP1/Yit and AKR/J strains. The production levels of IL‐10 and TGF‐β1 stimulated by LPS and CpG‐DNA were significantly lower in B cells separated from MLNs from the SAMP1/Yit strain. B cells expressing IL‐10 and TGF‐β1 were mainly located in a population characterized by the cell surface marker CD1d+. Interleukin‐1β production by TLR‐activated macrophages co‐cultured with MLN B cells from SAMP1/Yit mice was significantly higher than that of those from AKR/J mice. Interestingly, IFN‐γ production by T cells was noted only when they were co‐cultured with SAMP1/Yit but not the AKR/J B cells. These results are the first to show that disorders of regulatory B‐cell function under innate immune activation may cause disease pathogenesis in a murine model of Crohn’s disease.
Kinetic analysis of oxytetracycline residues in Chinese mitten crab,Eriocheir sinensis,muscle following intramuscular administration
Authors: Feng Q , Gao T , Ji H , Jiang X , Liang T , Gu W , Wu G , Gao G , Wang W
Published: June 2010

Crab culture is a very important economic industry in China. An epidemic of tremor disease of Chinese mitten crabs,Eriocheir sinensis,has become a serious problem in recent years. A spiroplasm has been proved to be the causative agent of this disease. Oxytetracycline (OTC) is used widely in aquaculture and was confirmed to be very effective against this pathogen. In this study, the distribution and depletion patterns of OTC in crab muscle were evaluated following single intramuscular doses of 2, 8 and 40 mg kg−1body weight. OTC was detected with a validated HPLC method. Concentration–time profiles were well described by a three‐compartment model with first‐order absorption after a single dose of 8 and 40 mg kg−1. For comparison, a non‐compartment model was employed. A withdrawal time of 48.29 and 55.92 days was suggested prior to consumption after receiving 8 and 40 mg kg−1. A recommended therapeutic dose of OTC in theory was calculated to be 36.37 mg kg−1. OTC was distributed well throughout the body. The elimination of OTC in muscle was slower compared with fish and other crustaceans. A dose of 40 mg kg−1is suggested for practical use.
The ABC‐transporter hutCD genes of Photobacterium damselae subsp. piscicida are essential for haem utilization as iron source and are expressed during infection in fish
Authors: Osorio C R , Juiz‐Río S , Lemos M L
Published: June 2010

The marine fish pathogen Photobacterium damselae subsp. piscicida utilizes haem compounds as the sole iron source. In a previous work, we characterized a gene cluster with ten potential haem uptake and utilization genes. Two of these genes, hutC and hutD, which are iron‐regulated, conform a putative inner membrane haem ABC transporter. In this study, we constructed an insertional mutant, leading to the inactivation of hutCD genes. Reverse transcriptase‐PCR analyses demonstrated that an insertion between the hutB and hutC genes abolished transcription of the downstream hutC and hutD genes. The hutCD mutant was unable to utilize haem as the sole iron source, demonstrating that the putative ABC‐transporter proteins HutC and HutD are essential for haem utilization as an iron source in P. damselae subsp. piscicida. In addition, reverse transcriptase‐PCR assays conducted with RNA samples isolated from experimentally infected fish revealed the presence of hutCD transcripts. The results demonstrate for the first time that haem uptake genes of a fish pathogen are expressed during the infective process in fish.
Antiparasitic activity of the antimicrobial peptide HbβP‐1, a member of the β‐haemoglobin peptide family
Authors: Ullal A J , Noga E J
Published: June 2010

A family of antimicrobial peptides (AMPs) derived from the β‐subunit of haemoglobin was recently isolated from channel catfish, Ictalurus punctatus, infected with Ichthyophthirius multifiliis (ich), an important freshwater fish parasite that causes ichthyophthiriosis. We previously discovered that one of these AMPs, HbβP‐1, had strong cidal activity against ich as well as another ectoparasite, Tetrahymena pyriformis. HbβP‐1 toxicity was specific, primarily affecting the trophozoite (trophont) stage of ich. Here, we show that HbβP‐1 acts more rapidly to kill smaller (presumably less mature) trophonts of ich, taking almost twice as long to kill larger trophonts (P < 0.0001). It acts more rapidly than an unrelated AMP, piscidin 1, which is haemolytic and also lethal to ich trophonts. HbβP‐1 is potently and selectively lethal to the trophont stage of the dinoflagellate ectoparasite, Amyloodinium ocellatum, one of the most important pathogens of warmwater marine fish. HbβP‐1 has no effect on the fish gill cell line feeder layer (G1B cells) used to propagate Amyloodinium, further suggesting a highly selective action. These findings suggest that HbβP‐1 or related AMPs might function in protecting marine as well as freshwater fish and that HbβP‐1 has highly selective activity against specific life stages of important fish ectoparasites.
A new distensibility technique to measure sphincter of Oddi function
Authors: Kunwald P. , Drewes A. M. , Kjær D. , Gravesen F. H. , Mcmahon B. P. , Madácsy L. , Funch‐jensen P. , Gregersen H.
Published: June 2010

Background  Evaluation of the biliary tract is important in physiological, pathophysiological, and clinical studies. Although the sphincter of Oddi (SO) can be evaluated with manometry, this technique has several limitations. This may explain the difficulties in identifying pathophysiological mechanisms for dysfunction of the SO and in identifying patients who may benefit from certain therapies. To encompass problems with manometry, methods such as the functional lumen imaging probe (FLIP) technique have been developed to study GI sphincters. This study set about miniaturising the FLIP probe and validating it for measurements in the SO. In order to get a better physiological understanding of the SO the aims were to show the sphincter profile in vivo and motility patterns of SO in pilot studies using volunteers that were experiencing biliary type pain but had normal SO manometry.

Methods  The SO probe was constructed to measure eight cross‐sectional areas (CSA) along a length of 25 mm inside a saline‐filled bag. To validate the technique for profiling the SO, six perspex cylinders with different CSAs were measured nine times to assess reproducibility and accuracy.

Key Results  Reproducibility and accuracy for these measurements were good. The probe performed well in bench tests and was therefore tested in four humans. The data indicated that it was possible to make distensions in the human SO and that a geometric sphincter profile could be obtained.

Conclusions & Inferences  The probe will in future studies be tested for diagnostic purposes related to sphincter of Oddi diseases.
Long‐term outcome of patients with lamivudine after early cessation of hepatitis B immunoglobulin for prevention of recurrent hepatitis B following liver transplantation
Authors: Yuefeng M. , Weili F. , Wenxiang T. , Ligang X. , Guiling L. , Hongwei G. , Wencai L. , Xiaoguang W. , Wei M. , Zhongyi F.
Published: June 2010

Yuefeng M, Weili F, Wengxiang T, Ligang X, Guiling L, Hongwei G, Wencai L, Xiaoguang W, Wei M, Zhongyi F. Long‐term outcome of patients with lamivudine after early cessation of hepatitis B immunoglobulin for prevention of recurrent hepatitis B following liver transplantation. Clin Transplant 2011: 25: 517–522. © 2010 John Wiley & Sons A/S.

Abstract: Background: The aim of this study is to examine the efficacy of long‐term prophylaxis with lamivudine (LAM) after a course of post‐operative hepatitis B immunoglobulin (HBIG) in patients who underwent liver transplantation (LT) for hepatitis B virus (HBV)‐related disease.

Result: The medical records of HBV‐infected patients who underwent a LT in our institution between July 2001 and May 2005 were reviewed. There were 15 liver transplant recipients who were administered HBIG for <18 months and used LAM as a maintenance prophylaxis regime enrolled in this study. At enrollment, all patients were hepatitis B surface antigen (HBsAg) positive and three patients were HBeAg positive. There were 13 patients who were HBV DNA positive with a mean viral load of 5.4 log copies/mL, and among them, 12 recipients were on antiviral therapy with LAM (100 mg/d orally) for 12–168 d, resulting in HBV DNA negative levels in nine patients prior to their transplant. HBV recurrence post‐LT was noted in two patients who had very high‐HBV DNA levels pre‐LT. Both of these patients showed LAM‐resistant mutation at the time of recurrence. The 11 patients who were HBV DNA negative before LT (low‐risk patients) had no HBV recurrence during a follow‐up at a median of 58 months post‐LT. This included five patients who had intermittent low‐level HBV DNA post‐LT (HBsAg negative), of whom two had YMDD mutation and these two were given adefovir in addition to LAM.

Conclusion: Our retrospective study demonstrated excellent long‐term outcomes in the low‐risk patients treated with LAM after a short course of HBIG.
Home air‐conditioning, traffic exposure, and asthma and allergic symptoms among preschool children
Authors: Zuraimi Mohamed Sultan , Tham Kwok‐Wai , Chew Fook‐Tim , Ooi Peng‐Lim , Koh David
Published: June 2010

To cite this article: Zuraimi MS, Tham K‐W, Chew F‐T, Ooi P‐L, Koh D. Home airconditioning, traffic exposure, and asthma and allergic symptoms among preschool children. Pediatr Allergy Immunol 2011; 22: e112–e118.

Epidemiological data suggest that traffic exposures can influence asthma and allergic symptoms among preschool children; however, there is no information on risk reduction via home air‐conditioning (AC). The aim of this study is to evaluate the associations of self‐reported traffic densities with asthma and allergic symptoms among preschool children and determine whether AC is an effect modifier. A cross‐sectional study adopting an expanded and modified ISAAC – International Study of Asthma and Allergies in Childhood conducted on randomly selected 2994 children living in homes without any indoor risk factors. Specific information on demographics, indoor home risk factors, and traffic variables were obtained. Adjusted prevalence ratios (PR) and 95% confidence interval (CI) were determined by Cox proportional hazard regression model with assumption of a constant risk period controlled for covariates. We found dose–response significant relationships between validated self‐reported traffic densities and asthma and rhinitis symptoms. Among children sleeping in non‐air‐conditioned homes, there were stronger associations between asthma and rhinitis symptoms studied. PRs for heavy traffic density were 2.06 for wheeze (95% CI 0.97–4.38), 2.89 for asthma (1.14–7.32), 1.73 for rhinitis (1.00–2.99), and 3.39 for rhinoconjunctivitis (1.24–9.27). There were no associations found for children sleeping in air‐conditioned homes. Our results suggest that AC in the bedroom modifies the health effects of traffic among preschool children. This finding suggests that attention should also be paid to ventilation characteristics of the homes to remediate health‐related traffic pollution problems.
Mitigation of the allergenic activity of beta‐lactoglobulin by electrolysis
Authors: Matsumoto Tomoaki
Published: June 2010

To cite this article: Matsumoto T. Mitigation of the allergenic activity of beta‐lactoglobulin by electrolysis. Pediatr Allergy Immunol 2011; 22: 235–242.

Beta‐lactoglobulin (BLG) is the most prominent allergen causing milk allergy and contains disulfide (S‐S) bonds that may be responsible for its allergic action. As S‐S bonds may be reduced during electrolysis, this study was undertaken to evaluate modulation of the allergic action of BLG after electrolysis. A current of 85 mA/cm2 was applied, and the allergic action was evaluated by means of competitive CAP‐FEIA inhibition tests and skin prick tests (SPT). Modification of BLG was examined by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐ToF‐MS). Although the number of total free sulfhydryl (SH) groups of BLG did not differ between the cathode and anode sides, BLG on the cathode side showed 54% lower inhibition than untreated BLG in the competitive CAP‐FEIA inhibition tests, and reduced wheal reactions, by 71%, in the SPT compared with those with untreated BLG. The SPT results with BLG on the anode side did not differ significantly from those with untreated BLG. The MALDI‐ToF‐MS results for the dimer of untreated BLG or BLG on the anode side showed two distinct peptide fragments (residues 41V‐60K and 149L‐162I) but, on the contrary, the dimer of BLG on the cathode side did not give these fragments, this being similar to in the case of the monomer of BLG. The allergic action of BLG was markedly mitigated during electrolysis on the cathode side, a dimer of BLG with a different mass spectrometric pattern from that of the dimer of untreated BLG being simultaneously formed.
Cuffed endotracheal tubes in children reduce sevoflurane and medical gas consumption and related costs
Authors: ESCHERTZHUBER S. , SALGO B. , SCHMITZ A. , ROTH W. , FROTZLER A. , KELLER C. H. , GERBER A. C. , WEISS M.
Published: June 2010

Background: This study aims to evaluate sevoflurane and anaesthetic gas consumption using uncuffed vs. cuffed endotracheal tubes (ETT) in paediatric surgical patients.

Methods: Uncuffed or cuffed ETT were used in paediatric patients (newborn to 5 years) undergoing elective surgery in a randomized order. Duration of assessment, lowest possible fresh gas flow (minimal allowed FGF: 0.5 l/min) and sevoflurane concentrations used were recorded. Consumption and costs for sevoflurane and medical gases were calculated.

Results: Seventy children (35 uncuffed ETT/35 cuffed ETT), aged 1.73 (0.01–4.80) years, were enrolled. No significant differences in patient characteristics, study period and sevoflurane concentrations used were found between the two groups. Lowest possible FGF was significantly lower in the cuffed ETT group [1.0 (0.5–1.0) l/min] than in the uncuffed ETT group [2.0 (0.5–4.3) l/min], P<0.001. Sevoflurane consumption per patient was 16.1 (6.4–82.8) ml in the uncuffed ETT group and 6.2 (1.1–14.9) ml in the cuffed ETT group, P=0.003. Medical gas consumption was 129 (53–552) l in the uncuffed ETT group vs. 46 (9–149) l in the cuffed ETT group, P<0.001. The total costs for sevoflurane and medical gases were 13.4 (6.0–67.3)€/patient in the uncuffed ETT group and 5.2 (1.0–12.5)€/patient in the cuffed ETT group, P<0.001.

Conclusions: The use of cuffed ETT in children significantly reduced the costs of sevoflurane and medical gas consumption during anaesthesia. Increased costs for cuffed compared with uncuffed ETT were completely compensated by a reduction in sevoflurane and medical gas consumption.
Root Growth Inhibition and Lignification Induced by Salt Stress in Soybean
Authors: Neves G. Y. S. , Marchiosi R. , Ferrarese M. L. L. , Siqueira‐Soares R. C. , Ferrarese‐Filho O.
Published: June 2010

Salt stress was evaluated on root growth, enzyme activities (phenylalanine ammonia‐lyase or PAL and soluble plus cell wall‐bound peroxidase or POD), hydrogen peroxide (H2O2) production, total phenolic content and lignin content and composition in soybean (Glycine max L. Merrill) roots. Three‐day‐old seedlings were cultivated in half‐strength Hoagland’s solution (pH 6.0), with or without addition of 50–200 mm of NaCl, into a growth chamber (25 °C, 12/12 h light/dark photoperiod, irradiance of 280 μmol m−2 s−1) for 24 h. In general, root length and fresh and dry weights decreased after NaCl treatment. PAL activity decreased, soluble and cell wall‐bound POD activities increased, and H2O2 content significantly decreased after NaCl exposure. Consequently, total phenolic and lignin contents and p‐hydroxyphenyl (H) and syringyl (S) monomers of lignin increased in NaCl‐treated roots. Altogether, these results suggest that the effects caused by NaCl may be owing to the enhanced lignin production that solidifies the cell wall and restricts root growth.
Initial evaluation of the forward‐viewing echoendoscope prototype for performing fine‐needle aspiration, Tru‐cut biopsy, and celiac plexus neurolysis
Authors: Eloubeidi Mohamad A
Published: June 2010

Background and Aims:  A forward‐viewing echoendoscope (FV‐CLA) has been recently developed for performing interventional endoscopic ultrasound (EUS). The role of FV‐CLA in performing standard EUS‐guided fine‐needle aspiration (FNA), Tru‐cut biopsy (TCB), and celiac plexus neurolysis (CPN) is unknown. Our aims were to evaluate the feasibility of the FV‐CLA for performing EUS‐guided FNA/TCB and CPN.

Methods:  In this prospective study conducted over a 3‐month period, 30 patients were evaluated with the FV‐CLA. Procedures performed were FNA in 28 lesions, TCB in one, and CPN in five patients.

Results:  EUS‐guided FNA was undertaken at the following sites: mediastinum (n = 3), liver (n = 2), retroperitoneal mass (n = 2), pancreas head/uncinate (n = 9), pancreas body (n = 6), pancreas tail (n = 4), and perigastric lymph node (n = 2). The median size of the lesions was 37 × 34 mm. A median of two passes was performed (range: 1–7). Final cytopathology diagnosed malignancies in 21 patients, with adenocarcinoma suspected for one.TCB of a mediastinal lymph node revealed lymphoma. FNA was benign in six patients. The sensitivity, specificity, positive predictive value, and negative predictive value for a malignancy diagnosis was 96% (95% confidence interval [CI], 87–96%), 100% (95% CI, 70–100%), 100% (92–100), and 86% (60–86%), respectively. CPN was successful in all five patients. It was easier to deploy the needle from the echoendoscope at all locations, including the duodenum, and irrespective of the site of the lesion.

Conclusions:  The initial evaluation and safety profile of the FV‐CLA echoendoscope for performing standard FNA/TCB and CPN appear to be favorable. The narrow image does not preclude basic therapeutic maneuvers. A major advantage appears to be easy needle deployment at any site within reach of the echoendoscope.
Clinical clues to suspicion of IgG4‐associated sclerosing cholangitis disguised as primary sclerosing cholangitis or hilar cholangiocarcinoma
Authors: Oh Hyoung‐Chul , Kim Myung‐Hwan , Lee Kyu Taek , Lee Jong Kyun , Moon Sung‐Hoon , Song Tae Jun , Eum Junbum , Park Do Hyun , Lee Sang Soo , Seo Dong‐Wan , Lee Sung Koo
Published: June 2010

Background and Aim:  This study aimed to determine the clinical characteristics of immunoglobulin G4 (IgG4)‐associated sclerosing cholangitis (ISC) and provide clinical clues differentiating ISC from primary sclerosing cholangitis (PSC) or hilar cholangiocarcinoma (CCC).

Methods:  Sixteen patients with ISC manifesting as hilar/intrahepatic strictures were analyzed for clinical characteristics and compared with patients with PSC and hilar CCC as disease controls for histology and serum IgG4 levels.

Results:  Distinguished biliary imaging findings of ISC included multifocal biliary tree involvement (n = 14), concentric bile duct thickening with preserved luminal patency (n = 13), and relatively mild proximal dilatation, despite prominent bile duct thickening (n = 11). Serum IgG4 levels were elevated in 12 patients (75%), but not in any of the 25 patients with hilar CCC. Ten patients (63%) had a past or concurrent history of autoimmune pancreatitis (AIP). The significant infiltration of IgG4‐positive cells was observed with endobiliary or liver biopsy in 11 of 16 patients (69%) with ISC, but not in any patients with PSC or hilar CCC. Extrabiliary organ involvement, including sialadenitis, inflammatory pseudotumor of the liver and kidney, and retroperitoneal fibrosis, was present in seven patients. Marked improvement of biliary strictures and/or extrabiliary involvement was observed in all ISC patients after steroid therapy.

Conclusions:  ISC should be considered in the differential diagnosis of hilar/intrahepatic biliary strictures. Past or concurrent AIP or extrabiliary organ involvement strongly suggests the possibility of ISC. Significant infiltration of IgG4‐positive cells on endobiliary or liver biopsy specimens, and/or elevated serum IgG4 levels, highly support the diagnosis of ISC and provide the rationale for steroid therapy.
Pathological bolus exposure plays a significant role in eliciting non‐cardiac chest pain
Authors: Kim Beom Jin , Choi Sung Chul , Kim Jae J , Rhee Jong Chul , Rhee Poong‐Lyul
Published: June 2010

Background and Aim:  Pathological bolus exposure is defined in the present study as cases in which all reflux percentage times are above 1.4% of the total reflux number, as revealed by impedance–pH monitoring. The role of pathological bolus exposure in the pathogenesis of non‐cardiac chest pain (NCCP) is poorly known. We aimed to classify and characterize NCCP using combined impedance–pH monitoring.

Methods:  Seventy‐five consecutive patients with NCCP were prospectively enrolled from January 2006 to October 2008. All the patients underwent upper endoscopy, esophageal manometry, and 24‐h multichannel intraluminal impedance (MII)–pH metering.

Results:  Sixteen patients (21.3%) had esophageal erosion upon endoscopy. Upon esophageal manometry, 37 patients (49.3%) had esophageal dysmotility. When the patients were classified based on MII–pH metering, 16 (21.3%) showed pathological acid exposure, and 40 (53.3%) showed pathological bolus exposure. The DeMeester score of patients with pathological acid exposure was higher than that of patients with pathological bolus exposure (P = 0.002). There was no significant difference in age, sex, typical esophageal symptoms, presence of esophageal erosion, esophageal dysmotility, improvement with proton pump inhibitor medication, symptom index ≥50%, percentage of time clearance pH below 4 ≥4%, and all reflux time ≥1.4% in the fasting period between the two groups. When the patients were divided into gastroesophageal reflux disease (GERD)‐related NCCP and non‐GERD‐related NCCP groups based on MII–pH metering and upper endoscopy, there was no difference between the two groups.

Conclusions:  Combined impedance–pH monitoring improves the detection and characterization of NCCP. This study suggests that pathological bolus exposure plays a major role in eliciting NCCP.