bluuebunny/crossref_metadata_2025 huggingface dataset:
Genetic and family structure in a group of 165 common bottlenose dolphins caught off the Japanese coast
Authors: Kita Yuki F. , Hosomichi Kazuyoshi , Suzuki Shingo , Inoko Hidetoshi , Shiina Takashi , Watanabe Mai , Tanaka Ayumi , Horie Taku , Ohizumi Hiroshi , Tanaka Sho , Iwasaki Toshihide , Ota Masao , Kulski Jerzy K.
Published: July 2012

Abstract

The biological and genetic structure of common bottlenose dolphins (Tursiops truncatus) that migrate seasonally near Japan remains largely unknown. We investigated the genetic and family structure in a group of 165 common bottlenose dolphins caught off the coast of Japan using mitochondrial DNA (mtDNA) and 20 microsatellite DNA markers. Phylogenetic analysis of the mtDNA control region sequences suggested that the dolphins were related more closely to oceanic types from Chinese waters than other geographic regions. The information on sex, sexual maturation and age together with the genetic markers revealed a strong likelihood for 37 familial relationships related mostly to maternity and an under‐representation of juvenile female offspring. The maternal dolphins had a similar offspring‐birth interval as the coastal types from North Atlantic Ocean, but a slightly younger first‐progeny age. The sex bias in the captured group was particularly marked towards an over‐representation of males among the young and immature dolphins, whereas the mature adults had an equal number of males and females. These results should be useful for future comparative biological, genetic and evolutionary investigations of bottlenose dolphins from the North Pacific Ocean with those from other regions.
Barrier dysfunction and bacterial uptake in the follicle‐associated epithelium of ileal Crohn's disease
Authors: Keita Åsa V. , Söderholm Johan D.
Published: June 2012

The ability to control uptake across the mucosa and protect from harmful substances in the gut lumen is defined as intestinal barrier function. The etiology of Crohn's disease is unknown, but genetic, environmental, and immunological factors all contribute. The frontline between these factors lies in the intestinal barrier. The most important inflammation‐driving environmental factor in Crohn's disease is the microbiota, whereEsherichia colistrains have been assigned a key role. The first observable signs of Crohn's disease are small aphtoid ulcers over Peyer's patches and lymphoid follicles. The overlaying follicle‐associated epithelium (FAE) is specialized for luminal sampling and is an entry site for antigens and bacteria. We have demonstrated increasedE. coliuptake across the FAE in Crohn's disease, which may initiate inflammation. This short review will discuss barrier dysfunction and bacteria in the context of ileal Crohn's disease, and how the FAE might be the site of initial inflammation.
Proof of concept for claudin‐targeted drug development
Authors: Suzuki Hidehiko , Kondoh Masuo , Takahashi Azusa , Yagi Kiyohito
Published: June 2012

Claudins (CLs) are a family of tetra‐integral membrane proteins that are a key structural and functional component of tight junctions. CLs are overexpressed in some malignant tumors. Claudin‐4 is highly expressed in the epithelial cells covering mucosal immune tissues. CLs may therefore be a potential target for improving drug absorption, treating cancer, and developing mucosal vaccine. Research using Clostridium perfringens enterotoxin has resulted in proofs of concept for CL‐targeted drug development. A platform for the creation of CL binders, such as immunization of CL and preparation of CL proteins, is now beginning to be established.
HIV infection and the intestinal mucosal barrier
Authors: Epple Hans‐Jörg , Zeitz Martin
Published: June 2012

HIV infection induces a barrier defect of the intestinal mucosa, which is closely linked to immune activation and CD4 T cell depletion. The HIV‐induced barrier defect is initiated in early acute and maintained through chronic infection. In acute infection, increased epithelial permeability is associated with increased epithelial apoptosis possibly caused by perforin‐expressing cytotoxic T cells. In chronic infection, mucosal production of inflammatory cytokines is associated with increased epithelial permeability, epithelial apoptosis, and alterations of epithelial tight junctions. In addition to HIV‐induced immune‐mediated effects, viral proteins have the potential to directly affect epithelial barrier function. After prolonged viral suppression by antiretroviral therapy, there is, at least partial, restoration of the HIV‐associated intestinal mucosal barrier defect despite persisting alterations of the mucosal immune system.
TNF‐α–induced intestinal epithelial cell shedding: implications for intestinal barrier function
Authors: Watson Alastair J.M. , Hughes Kevin R.
Published: June 2012

Although epithelial cells are continuously shed from the surface of the intestine, the intestinal epithelium maintains the integrity of the epithelial barrier. This is achieved through a highly dynamic process involving reorganization of tight junction and adherens junction proteins. This process both ejects the cell from the epithelial monolayer and plugs the gap left after the cell is shed. Inflammatory insults can trigger a disturbance of these barrier functions by increasing rates of cell shedding. Epithelial cell shedding and loss of barrier can be visualized by confocal laser endomicroscopy in humans. A simple grading system of confocal laser endomicroscopic images can stratify inflammatory bowel disease (IBD) patients in remission into those who will relapse over the subsequent six months and those who will not. Here, we review the mechanisms governing maintenance of these barrier functions and the implications of inflammation‐induced barrier dysfunction in IBD.
Myosin light chain kinase: pulling the strings of epithelial tight junction function
Authors: Cunningham Kevin E. , Turner Jerrold R.
Published: June 2012

Dynamic regulation of paracellular permeability is essential for physiological epithelial function, while dysregulated permeability is common in disease. The recent elucidation of the molecular composition of the epithelial tight junction complex has been accompanied by characterization of diverse intracellular mediators of paracellular permeabiltiy. Myosin light chain kinase (MLCK), which induces contraction of the perijunctional actomyosin ring through myosin II regulatory light chain phosphorylation, has emerged as a key regulator of tight junction permeability. Examination of the regulation and role of MLCK in tight junction dysfunction has helped to define pathological processes and characterize the role of barrier loss in disease pathogenesis, and may provide future therapeutic targets to treat intestinal disease.
A dynamic paracellular pathway serves diuresis in mosquito Malpighian tubules
Authors: Beyenbach Klaus W.
Published: June 2012

Female mosquitoes gorge on vertebrate blood, a rich nutrient source for developing eggs, but gorging meals increase the risk of predation. Mosquitoes are quick to reduce the flight payload with a potent diuresis. Diuretic peptides of the insect kinin family induce a tenfold reduction in the paracellular resistance of Malpighian tubules and increase the paracellular permeation of Cl−, the counterion of the transepithelial secretion of Na+ and K+. As a result, the transepithelial secretion of NaCl and KCl and water increases. Insect kinins signal the opening of the paracellular pathway via G protein–coupled receptors and the elevation of intracellular [Ca2+], which leads to the reorganization of the cytoskeleton associated with the septate junction (SJ). The reorganization may affect the septate junctional proteins that control the barrier and permselectivity properties of the paracellular pathway. The proteins involved in the embryonic formation of the SJ and in epithelial polarization are largely known for ectodermal epithelia, but the proteins that form and mediate the dynamic functions of the SJ in Malpighian tubules remain to be determined.
The yin and yang of claudin‐14 function in human diseases
Authors: Hou Jianghui
Published: June 2012

Claudins are tight junction integral membrane proteins that are key regulators of the paracellular pathway. The paracellular pathways in the inner ear and in the kidney are predominant routes for transepithelial cation transport. Mutations in claudin‐14 cause nonsyndromic recessive deafness DFNB29. A recent genome‐wide association study has identified claudin‐14 as a major risk gene of hypercalciuric nephrolithiasis. In vitro analyses show that claudin‐14 functions as a cation barrier in epithelial cells. The barrier function of claudin‐14 is crucial for generating the K+ gradient between perilymph and endolymph in the inner ear. However, neither homozygous individuals with DFNB29 mutations nor claudin‐14 knockout mice show any renal dysfunction. In this short review, I discuss several possible mechanisms to integrate the physiological function of claudin‐14 in the inner ear and the kidney.
The protozoan pathogen Toxoplasma gondii targets the paracellular pathway to invade the intestinal epithelium
Authors: Weight Caroline M. , Carding Simon R.
Published: June 2012

Abstract  Toxoplasma gondii is a ubiquitous parasite found within all mammals and birds worldwide that can cause fatal infections in immunocompromised persons and fetuses. The parasite causes chronic infections by residing in long‐living tissues of the muscle and brain. T. gondii infects the host through contaminated meat and water consumption with the gastrointestinal tract (GI tract) being the first point of contact with the host. The mechanisms by which the parasite invades the host through the GI tract are unknown, although it has been suggested that the paracellular pathway is important for parasite dissemination. Studies indicate that epithelial tight junction–associated proteins are affected by T. gondii, although which junctional proteins are affected and the nature of host protein–parasite interactions have not been established. We have uncovered evidence that T. gondii influences the cellular distribution of occludin to transmigrate the intestinal epithelium and suggest how candidate binding partners can be identified.
Zonulin, regulation of tight junctions, and autoimmune diseases
Authors: Fasano Alessio
Published: June 2012

Recent studies indicate that besides digestion and absorption of nutrients and water and electrolytes homeostasis, another key function of the intestine is to regulate the trafficking of environmental antigens across the host mucosal barrier. Intestinal tight junctions (TJs) create gradients for the optimal absorption and transport of nutrients and control the balance between tolerance and immunity to nonself antigens. To meet diverse physiological challenges, intestinal epithelial TJs must be modified rapidly and in a coordinated fashion by regulatory systems that orchestrate the state of assembly of the TJ multiprotein network. While considerable knowledge exists about TJ ultrastructure, relatively little is known about their physiological and pathophysiological regulation. Our discovery of zonulin, the only known physiologic modulator of intercellular TJs described so far, has increased our understanding of the intricate mechanisms that regulate the intestinal epithelial paracellular pathway and has led us to appreciate that its upregulation in genetically susceptible individuals leads to autoimmune diseases.
Calcium regulation of tight junction permeability
Authors: Bleich Markus , Shan Qixian , Himmerkus Nina
Published: June 2012

Calcium transport in the kidney is a key element in Ca2+ homeostasis. Ca2+ concentration, or more precisely the activity of freely dissociated Ca2+ ions, is a prerequisite for the appropriate function of virtually every cell. Along the renal tubule, about 85% of the filtered Ca2+ is transported across tight junctions at the paracellular route of reabsorption. Therefore, claudins, which form the conductive and selective part of the tight junctions, have moved into the focus of interest with respect to regulatory events in the control of Ca2+ transport. This control is of particular interest for the kidney since it has to defend itself against nephrocalcinosis and kidney stones. Tight junction proteins provide pathways, driving forces, and regulatory targets for Ca2+ transport. Direct regulation of tight junctions by changing Ca2+ concentrations allows fast and efficient feedback loops to adapt Ca2+ transport to the requirements of kidney function and plasma Ca2+ concentration.
SUMOylation of claudin‐2
Authors: Van Itallie Christina M. , Mitic Laura L. , Anderson James M.
Published: June 2012

The C‐terminal cytoplasmic tails of claudins are likely sites for interaction with proteins that regulate their function. We performed a yeast two‐hybrid screen with the tail of human claudin‐2 against a human kidney cDNA library and identified interactions with the PDZ3 domain of ZO‐2 as well as ubiquitin‐conjugating enzyme E2I (SUMO ligase‐1) and E3 SUMO‐protein ligase PIAS; the first is a predicted interaction, while the latter two are novel and suggest that claudin‐2 is a substrate for SUMOylation. Using an in vitro SUMOylation assay, we identified K218 as a conjugation site on claudin‐2; mutation of that lysine to arginine blocked SUMOylation. Stable expression of inducible GFP‐SUMO‐1 in MDCK cells resulted in decreased levels of claudin‐2 protein by immunoblot and decreased claudin‐2 membrane expression by immunofluorescence microscopy. We conclude that the cellular levels of claudin‐2 may be modulated by SUMOylation, warranting further investigation of cellular pathways that regulate this modification in vivo.
Impaired paracellular ion transport in the loop of Henle causes familial hypomagnesemia with hypercalciuria and nephrocalcinosis
Authors: Haisch Lea , Konrad Martin
Published: June 2012

Familial hypomagnesemia with hypercalciuria and nephrocalcinosis (FHHNC) is a rare tubular disorder caused by mutations in genes coding for tight junction (TJ) proteins. TJs define the paracellular path between adjacent cells and thereby play a pivotal role for the regulation of the paracellular ion permeability of epithelia. The family of TJ proteins comprise a variety of transmembrane proteins, including the claudins. Multiple distinct mutations in the genes for claudin‐16 and ‐19 have been described to be responsible for FHHNC. Both encoded proteins are especially important for the paracellular reabsorption of Mg2+ and Ca2+ in the thick ascending limb of Henle's loop. Interestingly, in addition to ion disturbances, FHHNC leads to chronic renal failure and may be associated with extrarenal symptoms.
Microbial butyrate and its role for barrier function in the gastrointestinal tract
Authors: Plöger Svenja , Stumpff Friederike , Penner Gregory B. , Schulzke Jörg‐Dieter , Gäbel Gotthold , Martens Holger , Shen Zanming , Günzel Dorothee , Aschenbach Joerg R.
Published: June 2012

Butyrate production in the large intestine and ruminant forestomach depends on bacterial butyryl‐CoA/acetate‐CoA transferase activity and is highest when fermentable fiber and nonstructural carbohydrates are balanced. Gastrointestinal epithelia seem to use butyrate and butyrate‐induced endocrine signals to adapt proliferation, apoptosis, and differentiation to the growth of the bacterial community. Butyrate has a potential clinical application in the treatment of inflammatory bowel disease (IBD; ulcerative colitis). Via inhibited release of tumor necrosis factor α and interleukin 13 and inhibition of histone deacetylase, butyrate may contribute to the restoration of the tight junction barrier in IBD by affecting the expression of claudin‐2, occludin, cingulin, and zonula occludens poteins (ZO‐1, ZO‐2). Further evaluation of the molecular events that link butyrate to an improved tight junction structure will allow for the elucidation of the cofactors affecting the reliability of butyrate as a clinical treatment tool.
Regulation of epithelial proliferation by tight junction proteins
Authors: Farkas Attila E. , Capaldo Christopher T. , Nusrat Asma
Published: June 2012

The epithelial tight junction (TJ) is the apical‐most intercellular junction and serves as a gatekeeper for the paracellular pathway by permitting regulated passage of fluid and ions while restricting movement of large molecules. In addition to these vital barrier functions, TJ proteins are emerging as major signaling molecules that mediate crosstalk between the extracellular environment, the cell surface, and the nucleus. Biochemical studies have recently determined that epithelial TJs contain over a hundred proteins that encompass transmembrane proteins, scaffolding molecules, cytoskeletal components, regulatory elements, and signaling molecules. Indeed, many of these proteins have defined roles in regulating epithelial polarity, differentiation, and proliferation. This review will focus on recent findings that highlight a role for TJ proteins in controlling cell proliferation during epithelial homeostasis, wound healing, and carcinogenesis.
The protein C pathway in intestinal barrier function: challenging the hemostasis paradigm
Authors: D’Alessio Silvia , Genua Marco , Vetrano Stefania
Published: June 2012

The protein C (PC) pathway is a well‐characterized anticoagulant system. Produced mainly by the liver as a zymogen, PC is activated on the vascular endothelial cell surface by thrombin–thrombomodulin complex. Once activated, PC inactivates two important cofactors of the coagulation cascade, factors Va and VIIIa, which are crucial for thrombin generation. For many years, this pathway has been studied for the clotting process, but only recently great progress has been made in understanding other functions of the PC system. Indeed, much work demonstrates that this pathway exerts several activities not only involved in the coagulative process but also in inflammation, cell proliferation, apoptosis, stabilization of endothelial barrier, and fibrinolysis. In addition, a recent study has shed light on a new role of the PC system in controlling intestinal permeability function by regulating tight junction molecules and promoting mucosal healing. This review highlights these recent insights in the context of the complex pathogenesis of inflammatory bowel disease.
Analysis of absorption enhancers in epithelial cell models
Authors: Rosenthal Rita , Heydt Miriam S. , Amasheh Maren , Stein Christoph , Fromm Michael , Amasheh Salah
Published: June 2012

A variety of chemical compounds are currently being discussed as novel drug delivery strategies. One promising strategy is to selectively open the paracellular pathway of epithelia for the passage of macromolecules. A prerequisite for this effect is a rapid and reversible action of these compounds, to allow a marked translocation of a drug, but also to avoid unwanted adverse effects, such as the translocation of noxious agents. Bioactive molecules that elevate paracellular permeability include Ca2+ chelators, bacterial toxins, and other compounds, some of which perturb the structural basis of epithelial barrier function—the tight junction. Within the tight junction, organ‐ and tissue‐specific barrier properties are determined mainly by claudins. The majority of members of the claudin protein family seal the paracellular pathway. This paper focuses on recent approaches concerning absorption‐enhancing effects, with regard to selectivity and mechanism.
Enteropathogenic E. coli effectors EspG1/G2 disrupt tight junctions: new roles and mechanisms
Authors: Glotfelty Lila G. , Hecht Gail A.
Published: June 2012

Enteropathogenic E. coli (EPEC) infection is a major cause of infantile diarrhea in the developing world. Using a type‐three secretion system, bacterial effector proteins are transferred to the host cell cytosol where they affect multiple physiological functions, ultimately leading to diarrheal disease. Disruption of intestinal epithelial cell tight junctions is a major consequence of EPEC infection and is mediated by multiple effector proteins, among them EspG1 and its homologue EspG2. EspG1/G2 contribute to loss of barrier function via an undefined mechanism that may be linked to their disruption of microtubule networks. Recently new investigations have identified additional roles for EspG. Sequestration of active ADP‐ribosylating factor (ARF) proteins and promotion of p21‐activated kinase (PAK) activity as well as inhibition of Golgi‐mediated protein secretion have all been linked to EspG. In this review, we examine the functions of EspG1/G2 and discuss potential mechanisms of EspG‐mediated tight junction disruption.
Defective tight junctions in refractory celiac disease
Authors: Schumann Michael , Kamel Sarah , Pahlitzsch Marie‐Luise , Lebenheim Lydia , May Claudia , Krauss Michael , Hummel Michael , Daum Severin , Fromm Michael , Schulzke Jörg‐Dieter
Published: June 2012

In celiac disease, the gut‐associated immune system is activated in response to the ingestion of gluten, causing an atrophy of the small intestinal mucosa. Although this condition is, in most cases, responsive to a gluten‐free diet, celiac disease refractory to treatment occurs in a small percentage of celiacs. An epithelial barrier defect is known to be an integral part of celiac pathophysiology. However, the mucosa in refractory celiac disease underlies a constant inflammatory process. The epithelial barrier has not been addressed in this condition so far. Herein, the tight junction‐associated barrier in refractory celiac disease is investigated functionally and structurally. Although normally expressed in celiac disease, claudin‐4 is shown to be downregulated in refractory cases, presumably by two mechanisms, reduced protein expression and increased claudin endocytosis. Furthermore, the tightening claudin‐5 is downregulated and the pore‐forming claudin‐2 is upregulated.
Claudin‐based paracellular proton barrier in the stomach
Authors: Tamura Atsushi , Yamazaki Yuji , Hayashi Daisuke , Suzuki Koya , Sentani Kazuhiro , Yasui Wataru , Tsukita Sachiko
Published: June 2012

The claudins comprise a multigene family that consists of at least 27 members. Claudins are responsible for establishing the paracellular barrier—which has permselectivity—at the tight junctions in epithelial cells, and the specific patterns of claudin expression in the epithelial cell sheets that cover the internal and external surfaces of organs contribute to the formation of microenvironments and organs’ biological functions. Data on the detailed characterization of individual claudins and their roles in different microenvironments are accumulating. A study on the stomach‐specific claudin‐18–knockout mouse, which has gastritis, recently revealed that the stomach‐type claudin‐18 specifically forms the proton barrier in the stomach, consistent with previously reported circumstantial evidence. Combined with previous studies on the specific ionic homeostasis by different types of claudins, our findings support the idea that claudins may regulate ion‐specific homeostasis in vivo.
Loss of enteral nutrition in a mouse model results in intestinal epithelial barrier dysfunction
Authors: Feng Yongjia , Ralls Matthew W. , Xiao Weidong , Miyasaka Eiichi , Herman Richard S. , Teitelbaum Daniel H.
Published: June 2012

Total parenteral nutrition (TPN) administration in a mouse model leads to a local mucosal inflammatory response, resulting in a loss of epithelial barrier function (EBF). Although, the underlying mechanisms are unknown, a major contributing factor is a loss of growth factors and subsequent critical downstream signaling. An important component of these is the p‐Akt pathway. An additional contributing factor to the loss of EBF with TPN is an increase in proinflammatory cytokine abundance within the mucosal epithelium, including TNF‐α and IFN‐γ. Loss of critical nutrients, including glutamine and glutamate, may affect EBF, contributing to the loss of tight junction proteins. Finding protective modalities for the small intestine during TPN administration may have important clinical applications. Supplemental glutamine and glutamate may be examples of such agents.
Effects of quercetin studied in colonic HT‐29/B6 cells and rat intestine in vitro
Authors: Amasheh Maren , Luettig Julia , Amasheh Salah , Zeitz Martin , Fromm Michael , Schulzke Jörg‐Dieter
Published: June 2012

The aim of this study was to analyze the influence of quercetin on intestinal barrier function using the human colonic epithelial cell line HT‐29/B6 and rat small and large intestine in vitro. Rat native ileum and late distal colon were incubated in Ussing chambers, and the total resistance (RT) was measured, and expression of tight junction proteins was characterized in immunoblots. By simulating inflammatory conditions with TNF‐α, we examined the barrier‐preventive effects of quercetin. Incubation with TNF‐α led to a decrease of RT in HT‐29/B6 cell monolayers, which could be partially inhibited by quercetin. In accordance with cell culture experiments, quercetin increased mucosal resistance of rat ileum and late distal colon. Thus, barrier disturbance in late distal colon specimens induced by TNF‐α and IFN‐γ could be partially prevented by coincubation with quercetin. These findings demonstrate that quercetin enhances barrier function in rat small and large intestine and possesses protective effects on cytokine‐induced barrier damage.
Abnormal intestinal permeability in Crohn's disease pathogenesis
Authors: Teshima Christopher W. , Dieleman Levinus A. , Meddings Jon B.
Published: June 2012

Increased small intestinal permeability is a longstanding observation in both Crohn's disease patients and in their healthy, asymptomatic first‐degree relatives. However, the significance of this compromised gut barrier function and its place in the pathogenesis of the disease remains poorly understood. The association between abnormal small intestinal permeability and a specific mutation in the NOD2 gene, which functions to modulate both innate and adaptive immune responses to intestinal bacteria, suggests a common, genetically determined pathway by which an abnormal gut barrier could result in chronic intestinal inflammation. Furthermore, rodent colitis models show that gut barrier defects precede the development of inflammatory changes. However, it remains possible that abnormal permeability is simply a consequence of mucosal inflammation. Further insight into whether abnormal barrier function is the cause or consequence of chronic intestinal inflammation will be crucial to understanding the role of intestinal permeability in the pathogenesis of Crohn's disease.
Tight junctions on the move: molecular mechanisms for epithelial barrier regulation
Authors: Shen Le
Published: June 2012

Increasing evidence suggests that the tight junction is a dynamically regulated structure. Cytoskeletal reorganization, particularly myosin light chain phosphorylation‐‐induced actomyosin contraction, has increasingly been recognized as a mediator of physiological and pathophysiological tight junction regulation. However, our understanding of molecular mechanisms of tight junction modulation remains limited. Recent studies using live cell and live animal imaging techniques allowed us to peek into the molecular details of tight junction regulation. At resting conditions, the tight junction is maintained by dynamic protein–protein interactions, which may provide a platform for rapid tight junction regulation. Following stimulation, distinct forms of tight junction protein reorganization were observed. Tumor necrosis factor (TNF‐α) causes a myosin light chain kinase (MLCK)‐‐mediated barrier regulation by inducing occludin removal from the tight junction through caveolar endocytosis. In contrast, MLCK‐ and CK2‐inhibition‐‐caused tight junction regulation is mediated by altered zonula occludens (ZO)‐1 protein dynamics and requires ZO‐1‐‐mediated protein–protein interaction, potentially through regulating claudin function. Although some of the molecular details are missing, studies summarized above point to modulating protein localization and dynamics that are common mechanisms for tight junction regulation.
Ion transport and barrier function are disturbed in microscopic colitis
Authors: Barmeyer Christian , Erko Irene , Fromm Anja , Bojarski Christian , Allers Kristina , Moos Verena , Zeitz Martin , Fromm Michael , Schulzke Jörg‐Dieter
Published: June 2012

In this paper, we identify mechanisms of watery diarrhea in microscopic colitis (MC). Biopsies from the sigmoid colon of patients with collagenous colitis and treated lymphocytic colitis were analyzed in miniaturized Ussing chambers for electrogenic sodium transport and barrier function with one‐path impedance spectroscopy. Cytometric bead arrays (CBA) served to analyze cytokine profiles. In active MC, electrogenic sodium transport was diminished and epithelial resistance decreased. CBA revealed a Th1 cytokine profile featuring increased IFN‐γ, TNF‐α, and IL‐1β levels. After four weeks of steroid treatment with budesonide, electrogenic sodium transport recovered while epithelial barrier defects remained. Diarrhea in MC results at least in part from a combination of impaired electrogenic sodium transport and barrier defects. From a therapeutic perspective it can be postulated that the functional importance of loss of ions may be higher than that caused by barrier impairment.
Comparison of Health‐Relevant Flavonoids in Commonly Consumed Cranberry Products
Authors: Grace Mary H. , Massey Aaron R. , Mbeunkui Flaubert , Yousef Gad G. , Lila Mary Ann
Published: July 2012

Abstract:  The human health benefits from consumption of cranberry products have been associated with the fruits’ unique flavonoid composition, including a complex profile of anthocyanins and proanthocyanidins. However, when processed by techniques such as pressing, canning, concentrating, or drying, a number of these natural components may be compromised or inactivated due to physical separation, thermal degradation, or oxidation. Fresh cranberries were compared to freeze‐dried berries and individual fruit tissues (skin and peeled fruit). Products examined included cranberry juices (commercial and prepared from concentrate), cranberry sauces (commercial and homemade), and sweetened‐dried cranberries (commercial). Freeze‐drying resulted in no detectable losses of anthocyanins or proanthocyanidins from cranberry fruits. Anthocyanins were localized in the skin. Proanthocyanins were higher in the skin than in the flesh, with the exception of procyanidin A‐2 dimer which was concentrated in the flesh. Anthocyanins were significantly higher in not‐from‐concentrate juice than in reconstituted juice from concentrate (8.3 mg and 4.2 mg/100 mL, respectively). Similarly, proanthocyanidins were markedly higher in not‐from‐concentrate juice compared to juice from concentrate (23.0 mg and 8.9 mg/100 mL, respectively). Homemade sauce contained far higher anthocyanins and proanthocyanidins (15.9 and 87.9 mg/100 g, respectively) than canned sauces processed with whole berries (9.6 and 54.4 mg/100 g, respectively) or jelled‐type (1.1 and 16 mg/100 g, respectively). Sweetened‐dried cranberries were quite low in anthocyanins (7.9 mg/100 g), but they still retained considerable proanthocyanidins (64.2 mg/100 g). Commercially processed products contained significantly lower levels of polyphenols as compared to fresh and home‐processed preparations. Anthocyanins were more sensitive to degradation than proanthocyanidins.

Practical Application:  As cranberry juices and other products are increasingly consumed for their recognized health benefits (including prophylaxis against urinary tract infection), it is relevant to consider how various degrees of commercial and home processing can alter innate levels of the biologically active flavonoids (especially anthocyanins and proanthocyanidins) characteristic to the intact fruits.
Motor neurone disease in molybdenum‐deficient sheep fed the endogenous purine xanthosine: possible mechanism for Tribulus staggers
Authors: Bourke CA
Published: June 2012

Background  The occurrence of Tribulus terrestris motor neurone disease (MND) in sheep is linked with grazing Tribulus growing on cultivation paddocks. A previous survey found that the molybdenum (Mo) content of Tribulus growing on uncultivated soils in the Coonabarabran district of New South Wales was 3.03 ppm, but on cultivated soils it was <0.04 ppm. Tribulus contains the purine, xanthosine, which functions as a neuromodulator, and the catabolism of xanthosine is Mo‐dependent.

Design  To investigate the relationship between xanthosine ingestion and low Mo concentration, eight sheep were fed Mo‐deficient lucerne chaff (<0.10 ppm), the Mo antagonist, sodium tungstate, and xanthosine (25 mg/kg/day) over 18 weeks and then returned to pasture.

Results  Signs of MND developed in two sheep 30 months later and astrocyte degeneration occurred in all sheep.

Conclusion  The findings were similar to those observed in sheep with T. terrestris MND, suggesting that the combination of xanthosine ingestion and Mo deficiency may be the cause of this disorder.
Murray Valley encephalomyelitis in a horse
Authors: Holmes JM , Gilkerson JR , El Hage CM , Slocombe RF , Muurlink MA
Published: June 2012

A 5‐year‐old Thoroughbred mare presented with signs of severe pain and was taken to exploratory laparotomy based on suspicion of an acute abdominal lesion. A mild gastrointestinal lesion was discovered, but was considered disproportional to the severity of signs displayed. The mare was later euthanased because of intractable pain. Comprehensive postmortem examination, including polymerase chain reaction testing of central nervous system tissue samples, allowed a definitive diagnosis of Murray Valley encephalomyelitis to be made. This case demonstrates the variability of clinical presentations in horses infected with Murray Valley encephalitis virus.
Plastic reduction for anatomical gingival fibromatosis
Authors: Ribeiro‐Júnior O. , Borba A.M. , Lemos‐Júnior C.A. , Naclério‐Homem M.G. , Kolokythas A.
Published: July 2012

Aim:  Anatomical gingival fibromatosis is an uncommon process involving the palatal gingival and maxillary tuberosity generally in a bilateral and symmetrical manner. Treatment varies from basic periodontal care to surgical removal when the condition causes difficulties with oral hygiene, is associated with secondary periodontal diseases or interferes with prosthesis stability. The authors present a plastic reduction technique modified for tooth‐bearing areas and discuss its application.

Material and methods:  The proposed technique was accomplished in three patients presenting with anatomical gingival fibromatosis. Surgery consisted of a three‐step technique that aimed at the reduction of the fibromatosis and the reconstruction of the palatal gingiva.

Results:  None of the patients present post‐operative complications during a follow‐up period that varied from 6 to 12 months. After surgery all patients improved their ability to provide proper hygiene for the affected area as well as their periodontal condition.

Conclusions:  The present surgical technique is a treatment option with a good result while maintaining periodontal health.
Dentists' attitudes, behaviors, and barriers related to tobacco‐use cessation in the dental setting
Authors: Prakash Preeti , Belek Marilynn G. , Grimes Barbara , Silverstein Steven , Meckstroth Richard , Heckman Barbara , Weintraub Jane A. , Gansky Stuart A. , Walsh Margaret M.
Published: June 2012

Objective: This study assessed attitudes, behaviors, and barriers among general dentists in California, Pennsylvania, and West Virginia, related to patient tobacco cessation counseling.

Methods: From 2004 to 2008, a baseline survey was mailed to 271 study dentists randomly selected from a master Delta Dental Insurance Company provider list in each state who had agreed to participate in a tobacco cessation randomized clinical trial. Four backward logistic regression models assessed correlates of the five As related to tobacco cessation: Asking about tobacco use, Advising users to quit, Assessing readiness to quit, Assisting with quitting, and Arranging follow‐up.

Results: Most respondents (n = 265) were male, had practiced dentistry for over 15 years, asked about tobacco use (74%), and advised tobacco users to quit (78%). Only 19% assessed readiness to quit; 39% assisted with quitting; 4% arranged follow‐up; and 42% had formal training in tobacco cessation. Believing that tobacco cessation counseling was an important professional responsibility, practicing <15 years, and asking about tobacco use significantly related to advising users to quit. Providing cessation advice and feeling effective intervening related to assessing readiness to quit. Advising users to quit, assessing readiness to quit, feeling effective intervening, and having had formal tobacco cessation training related to assisting with quitting. Barriers to cessation counseling were perceived patient resistance (66%), lack of insurance reimbursement (56%), not knowing where to refer (49%), and lack of time (32%).

Conclusion: Study dentists reported not fully performing the five As. Advising, assessing, having formal training, and feeling effective increased the likelihood of cessation counseling.
Using chlorhexidine varnish to prevent early childhood caries in American Indian children
Authors: Robertson Lawrence D. , Phipps Kathy R. , Oh Junhie , Loesche Walter J. , Kaciroti Niko , Symington John M.
Published: June 2012

Objectives: To test the efficacy of 10% chlorhexidine (CHX) dental varnish applied to the mothers' dentition in preventing caries in American Indian children.

Methods: This was a placebo‐controlled, double‐blind, randomized clinical trial. Mother–child pairs were enrolled when the child was 4.5‐6.0 months. Mothers received 4 weekly applications of the study treatment (CHX or placebo) followed by single applications when her child was age 12 and 18 months. Children received caries examinations at enrollment, 12, 18 and 24 months. Analyses were limited to the intent‐to‐treat (ITT) group: children whose mothers received the first study treatment and who received at least one post‐baseline exam. The outcome variable was the number of new carious surfaces (NNCS) at the child's last visit. Wilcoxon nonparametric and Fisher's exact tests were used to test differences between the active and placebo groups.

Results: We randomized 414 mother–child pairs, with 367 (88.6%) included in the ITT group (active = 188, placebo = 179). The proportion of children caries‐free at their final exam was 51.1% and 50.8% for the active and placebo groups (P > 0.99). The mean NNCS for the active and placebo groups was 3.82 (standard deviation [SD] = 8.18) and 3.80 (SD = 6.08), respectively (P = 0.54). The proportion with NNCS > 6 was 18.1% for active children versus 27.9% for placebo (relative risk [RR] = 0.65, P = 0.03). The number needed to treat to shift one child from NNCS > 6 to a lower severity was 10.2.

Conclusions: In this population CHX varnish did not reduce the mean NNCS or proportion of children with caries, but did reduce the proportion with severe caries.
IVC filter limb penetration of the caval wall during retroperitoneal surgery/lymph node dissection
Authors: Goh Gerard S , Slattery Michael M , Given Mark F , Little David , Lee Michael J
Published: June 2012

Optional inferior vena cava (IVC) filters are being increasingly used for protection against pulmonary embolism in patients with deep vein thrombosis where anticoagulation is contraindicated. We describe two cases during retroperitoneal surgery where the IVC filters were found to have perforated the cava wall and were subsequently removed intra‐operatively. Cava wall penetration by filter limbs poses a significant danger during retroperitoneal lymph node dissection and filters should be removed preoperatively.
Extra‐medullary haematopoiesis: A pictorial review of its typical and atypical locations
Authors: Sohawon Dharamraj , Lau Kenneth K , Lau Theodore , Bowden Donald K
Published: June 2012

Extra‐medullary haematopoiesis (EMH) occurs in many haematological disorders and is secondary to insufficient bone marrow function or ineffective erythropoiesis. It usually presents as haematopoietic masses in several typical and atypical body locations. This pictorial review briefly discusses the common EMH locations encountered in clinical practice, including the liver, spleen, lymph nodes and paravertebral regions. Unusual presentation as focal hepatic and splenic masses is also discussed. Some atypical EMH locations are then presented together with their pathophysiology and relevant illustrations. These include EMH in the intra‐spinal canal, pre‐sacral region, nasopharynx and paranasal sinuses. Intra‐spinal EMH can cause cord compression and present with neurological symptoms. In these cases, urgent treatment may be required. Haematopoietic masses in the other atypical locations can present with obstructive symptoms or may be diagnosed incidentally on imaging. EMH in unusual locations need to be monitored with follow‐up imaging to ensure their stability. In some circumstances, tissue biopsy is appropriate to confirm the diagnosis and exclude other sinister pathology (e.g. malignancy). Treatment is only required where symptoms are present. Management options include blood transfusion, radiotherapy, hydroxyurea or surgical resection in selected cases.
Case of bilateral non‐traumatic subperiosteal orbital haematomas
Authors: Lee Daniel K , Tran Phillip V , Lau Kenneth K
Published: June 2012

Subperiosteal orbital haematoma is an uncommon entity, usually unilateral and caused by trauma. We present a unique case of bilateral non‐traumatic subperiosteal orbital haematomas caused by thrombolysis and anticoagulation.
Haemobilia – a rare presentation of intrabiliary hydatid disease
Authors: Lakshminarayan Raghuram , Anuradha Chandramohan
Published: June 2012

Intrabiliary hydatid cysts have been known to cause complications. We report a rare case of calcified crumpled intrabiliary hydatid cyst causing massive haemobilia due to a hepatic artery pseudoaneurysm, an unusual complication of intrabiliary hydatid cyst. The patient was successfully treated by stenting of the pseudoaneurysm with overlapping stents.
The effectiveness and evaluation of conservation planning
Authors: Bottrill Madeleine C. , Pressey Robert L.
Published: July 2012

Investment of time and resources in conservation planning has grown exponentially in recent years; yet there has been limited evaluation of the benefits and costs of investing in planning exercises. Without evaluation, investments in planning are not accountable, decisions are not defensible, and learning from past experiences is limited. Bringing together information from published literature, planning documents, and new qualitative data from interviews with planners, we describe an evaluation framework for conservation planning to more fully address the potential range of outcomes, categorized across five types of capital: natural, financial, social, human, and institutional. We review the extent to which evidence supports these types of outcomes, and finish by considering the conceptual, operational, organizational, and policy implications of improved evaluation in planning. If conservation planning seeks to be effective, adaptive, and informative, then systems of evaluation must be considered from the outset of planning processes to support learning and improvement of outcomes.
Testing four candidate barcoding markers in temperate woody bamboos (Poaceae: Bambusoideae)
Authors: CAI Zhao‐Ming , ZHANG Yu‐Xiao , ZHANG Li‐Na , GAO Lian‐Ming , LI De‐Zhu
Published: June 2012

Abstract  Bambusoideae is an important subfamily of the grass family Poaceae that has considerable economic, ecologic and cultural value. In addition, Bambusoideae species are important constituents of the forest vegetation in China. Because of the paucity of flower‐bearing specimens and homoplasies of morphological characters, it is difficult to identify species of Bambusoideae using morphology alone, especially in the case of temperate woody bamboos (i.e. Arundinarieae). To this end, DNA barcoding has shown great potential in identifying species. The present study is the first attempt to test the feasibility of four proposed DNA barcoding markers (matK, rbcL, trnH–psbA, and internal transcribed spacer [ITS]) in identifying 27 species of the temperate woody bamboos. Three plastid markers showed high levels of universality, whereas the universality of ITS was comparatively low. A single plastid marker provided low levels of discrimination success at both the genus and species levels (<12%). Among the combinations of plastid markers, the highest discriminatory power was obtained using the combination of rbcL+matK (14.8%). Using a combination of three markers did not increase species discrimination. The nuclear region ITS alone could identify 66.7% of species, although fewer taxa were included in the ITS analyses than in the plastid analyses. When ITS was integrated with a single or combination of plastid markers, the species discriminatory power was significantly improved. We suggest that a combination of rbcL+ ITS, which exhibited the highest species identification power of all combinations in the present study, could be used as a potential DNA barcode for temperate woody bamboos.
Increased glutathione and mitogen‐activated protein kinase phosphorylation are involved in the induction of doxorubicin resistance in hepatocellular carcinoma cells
Authors: Ye Cai‐Guo , Yeung John Hok‐Keung , Huang Guo‐Liang , Cui Ping , Wang Jian , Zou Yin , Zhang Xiang‐Ning , He Zhi‐Wei , Cho Chi‐Hin
Published: June 2012

Aim:  The human hepatocellular carcinoma (HCC) cell line HepG2 can easily acquire resistance to doxorubicin. However, the mechanism of action is unclear.

Methods:  In the present study, we used confocal microscopy, flow cytometry and other methods to reveal the mechanisms by which HepG2 cells acquire doxorubicin resistance.

Results:  Our results showed that R‐HepG2 cells, a doxorubicin‐resistant sub‐line of HepG2, exhibited decreased intracellular accumulation of doxorubicin and increased expression of P‐glycoprotein (P‐gp) and multidrug resistance‐associated protein 1 when compared with HepG2 cells. R‐HepG2 cells also harbored higher levels of glutathione and increased expression of glutathione peroxidase. Furthermore, we demonstrated that the phosphorylation of mitogen‐activated protein kinases (p38 and c‐jun‐N‐terminal kinases), IkBα and CREB were increased in R‐HepG2 cells. Specific p38 inhibitor SB203580 decreased P‐gp expression. The multi‐kinase inhibitor sorafenib tosylate also significantly suppressed the phosphorylation of these proteins and inhibited the expression of P‐gp.

Conclusion:  These findings reveal that the drug resistance could be acquired through mitogen‐activated protein kinase‐dependent upregulation of P‐gp. This mechanism protects R‐HepG2 cells from the anticancer action of doxorubicin.